BR122023011393B1 - COMPOUND, PHARMACEUTICAL COMPOSITION AND USES OF A COMPOUND - Google Patents

Abstract

The disclosure presents macrocyclic compounds and pharmaceutical compositions and protein complexes thereof, capable of inhibiting Ras proteins and their uses in the treatment of cancers.

Description

CROSS-REFERENCE TO RELATED ORDERS

[001] This application claims the benefit of priority of U.S. Application No. 62/930,355, filed November 4, 2019; U.S. Application No. 62/951,652, filed December 20, 2019; U.S. Application No. 63/000,357, filed March 26, 2020; U.S. Application No. 63/011,636, filed April 17, 2020; and U.S. Application No. 63/043,588, filed June 24, 2020, all of which are incorporated herein by reference in their entirety.

FUNDAMENTALS

[002] The vast majority of small molecule drugs act by binding to a functionally important pocket on a target protein, thereby modulating the activity of that protein. For example, the cholesterol-lowering drugs known as statins bind to the enzymatic active site of HMG-CoA reductase, thereby preventing the enzyme from engaging its substrates. The fact that many such drug/target interaction pairs are known may have led some to believe that a small molecule modulator could be discovered for most, if not all, proteins given a reasonable amount of time, effort, and resources. This is far from the case. Current estimates are that only about 10% of all human proteins are targetable by small molecules. Bojadzic and Buchwald, Curr Top Med Chem 18: 674–699 (2019). The other 90% are currently considered refractory or intractable for the aforementioned small molecule drug discovery. Such targets are commonly referred to as “undruggable.” These undruggable targets include a vast and largely unexplored reservoir of clinically important human proteins. Thus, there is great interest in discovering new molecular modalities capable of modulating the function of such undruggable targets.

[003] It has been well established in the literature that Ras proteins (KRas, H-Ras, and N-Ras) play an essential role in several human cancers and are therefore appropriate targets for anticancer therapy. Indeed, mutations in Ras proteins account for approximately 30% of all human cancers in the United States, many of which are fatal. Dysregulation of RAS proteins by activating mutations, overexpression, or upstream activation is common in human tumors, and activating mutations in RAS are frequently found in human cancer. For example, activating mutations at codon 12 in Ras proteins function by inhibiting the rates of intrinsic and GTPase-activating protein (GAP)-dependent hydrolysis of GTP, significantly biasing the population of mutant Ras proteins toward the “on” (GTP-bound) (Ras(ON)) state, leading to oncogenic MAPK signaling. Notably, RAS exhibits a picomolar affinity for GTP, allowing RAS to be activated even in the presence of low concentrations of this nucleotide. Mutations in codons 13 (e.g., G13D) and 61 (e.g., Q61K) of Ras are also responsible for oncogenic activity in some cancers.

[004] Despite extensive drug discovery efforts against RAS over the past decades, a drug directly targeting RAS has not yet been approved. Additional efforts are needed to discover additional drugs for cancers driven by the various RAS mutations.

SUMMARY

[005] Provided herein are inhibitors of Ras. The approach described herein involves the formation of a high-affinity three-component complex, or conjugate, between a synthetic ligand and two intracellular proteins that do not interact under normal physiological conditions: the target protein of interest (e.g., Ras) and a widely expressed cytosolic chaperone (presenting protein) in the cell (e.g., cyclophilin A). More specifically, in some embodiments, the Ras inhibitors described herein induce a novel binding pocket in Ras, leading to the formation of a high-affinity triple complex, or conjugate, between the Ras protein and the widely expressed cytosolic chaperone, cyclophilin A (CYPA). Without being bound by theory, the inventors believe that one way in which the inhibitory effect on Ras is effected by the compounds of the invention and the complexes, or conjugates, they form is by steric occlusion of the interaction site between Ras and downstream effector molecules, such as RAF and PI3K, which are necessary to propagate the oncogenic signal.

[006] As such, in some embodiments, the disclosure features a compound, or pharmaceutically acceptable salt thereof, of structural Formula I: Formula Iwhere the dotted lines represent zero, one, two, three, or four nonadjacent double bonds;A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 10-membered heteroarylene;B is absent, is -CH(R9)-, >C=CR9R9', or >CR9R9' where the carbon is bonded to the carbonyl carbon of -N(R11)C(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;G is optionally substituted C1-C4 alkylene, Optionally substituted C1-C4 alkenylene, optionally substituted C1-C4 heteroalkylene, -C(O)O-CH(R6)- where C is attached to -C(R7R8)-, -C(O)NH-CH(R6)- where C is attached to -C(R7R8)-, optionally substituted C1-C4 heteroalkylene, or 3- to 8-membered heteroarylene;L is absent or is a ligand;W is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone, a haloacetal, or an alkynyl sulfone;X1 is optionally substituted C1-C2 alkylene, NR, O, or S(O)n;X2 is O or NH;X3 is N or CH;n is 0, 1, or 2;R is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2C4 alkenyl, C2-C4 optionally substituted alkynyl, C(O)R', C(O)OR', C(O)N(R')2, S(O)R', S(O)2R', or S(O)2N(R')2; each R' is independently H or optionally substituted C1-C4 alkyl; Y1 is C, CH, or N; Y2, Y3, Y4, and Y7 are independently C or N; Y5 is CH, CH2, or N; Y6 is C(O), CH, CH2, or N; R1 is cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl, or R1 and R2 combine with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl; R2 is absent, is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl; R3 is absent, or R2 and R3 combine with the atom to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or optionally substituted 3- to 14-membered heterocycloalkyl; R4 is absent, is hydrogen, halogen, cyano, or methyl optionally substituted by 1 to 3 halogens; R5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl, or cyclobutyl; R6 is hydrogen or methyl; R7 is hydrogen, halogen, or optionally substituted C1-C3 alkyl, or R6 and R7 combine with the carbon atoms to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7 and R8 combine with the carbon atom to which they are attached to form C=CR7'R8'; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;R7a and R8a are independently hydrogen, halo, C1-C3 alkyl, or combine with the carbon to which they are attached to form a carbonyl;R7' is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R8' is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7' and R8' combine with the carbon atom to which they are attached to form optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R9 is H, F optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; or R9 and L combine with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl; R9' is hydrogen or optionally substituted C1-C6 alkyl; or R9 and R9', combined with the atoms to which they are attached, form a 3- to 6-membered cycloalkyl or a 3- to 6-membered heterocycloalkyl; R10 is hydrogen, halo, hydroxy, C1-C3 alkoxy, or C1-C3 alkyl; R10a is hydrogen or halo; R11 is hydrogen or C1-C3 alkyl; and R21 is hydrogen or C1-C3 alkyl (e.g., methyl).

[007] Also provided are pharmaceutical compositions comprising a compound of Formula I, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[008] Further provided is a conjugate, or salt thereof, comprising the structure of Formula IV:ML-PFormula IV wherein L is a ligand; P is a monovalent organic moiety; and M has the structure of Formula V Formula It follows that the dotted lines represent zero, one, two, three or four non-adjacent double bonds; A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 6-membered heteroarylene;B is absent, is -CH(R9)-, >C=CR9R9', or >CR9R9' where the carbon is bonded to the carbonyl carbon of -N(R11)C(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;G is optionally substituted C1-C4 alkylene, optionally substituted C1-C4 alkenylene, optionally substituted C1-C4 heteroalkylene, - C(O)O-CH(R6)- where C is attached to -C(R7R8)-, -C(O)NH-CH(R6)- where C is attached to -C(R7R8)-, optionally substituted C1-C4 heteroalkylene, or 3- to 8-membered heteroarylene;X1 is optionally substituted C1-C2 alkylene, NR, O, or S(O)n;X2 is O or NH;X3 is N or CH;n is 0, 1, or 2;R is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2C4 alkenyl, optionally substituted C2-C4 alkynyl, C(O)R', C(O)OR', C(O)N(R')2, S(O)R', S(O)2R', or S(O)2N(R')2;each R' is independently H or C1-C4 alkyl optionally substituted;Y 1 is C, CH, or N;Y 2, Y3, Y4, and Y7 are independently C or N;Y 5 is CH, CH2, or N;Y 6 is C(O), CH, CH2, or N; R1 is cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl, or R1 and R2 combine with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl; R2 is absent, is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl; R3 is absent, or R2 and R3 combine with the atom to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or optionally substituted 3- to 14-membered heterocycloalkyl; R4 is absent, is hydrogen, halogen, cyano, or methyl optionally substituted by 1 to 3 halogens; R5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl, or cyclobutyl; R6 is hydrogen or methyl; R7 is hydrogen, halogen, or optionally substituted C1-C3 alkyl, or R6 and R7 combine with the carbon atoms to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7 and R8 combine with the carbon atom to which they are attached to form C=CR7'R8'; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;R7a and R8a are independently hydrogen, halo, C1-C3 alkyl, or combine with the carbon to which they are attached to form a carbonyl;R7' is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R8' is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7' and R8' combine with the carbon atom to which they are attached to form optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R9 is H, F optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl, or R9 and L combine with the atoms to which they are attached to form form an optionally substituted 3- to 14-membered heterocycloalkyl; R9' is hydrogen or optionally substituted C1-C6 alkyl; or R9 and R9', combined with the atoms to which they are attached, form a 3- to 6-membered cycloalkyl or a 3- to 6-membered heterocycloalkyl; R10 is hydrogen, halo, hydroxy, C1-C3 alkoxy, or C1-C3 alkyl; R10a is hydrogen or halo; R11 is hydrogen or C1-C3 alkyl; and R21 is H or C1-C3 alkyl.

[009] Also provided is a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof.

[0010] In some embodiments, a method of treating a Ras protein-related disorder in a subject in need thereof is provided, the method comprising administering to the subject a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof.

[0011] Further provided is a method of inhibiting a Ras protein in a cell, the method comprising contacting the cell with an effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof.

[0012] It is specifically contemplated that any limitation discussed with respect to one embodiment of the invention may apply to any other embodiment of the invention. Furthermore, any compound or composition of the invention may be used in any method of the invention, and any method of the invention may be used to produce or utilize any compound or composition of the invention.

BRIEF DESCRIPTION OF THE DRAWINGS

[0013] FIG. 1A and FIG. 1B: These figures illustrate a matched pair analysis of potencies of certain compounds of the present invention (Formula BB) (right dots) and corresponding compounds of Formula AA (left dots) in which one H is replaced by (S)Me in the context of two different cell-based assays. The y-axes represent pERK EC50 (FIG. 1A) or CTG IC50 (FIG. 1B) as measured in an H358 cell line.

[0014] FIG. 2A and FIG. 2B: A compound of the present invention, Compound A, led to profound regressions in vivo in an NSCLC (KRAS G12C) xenograft model. Some animals exhibited complete responses (CR) = 3 consecutive tumor measurements < 30 mm3. FIG. 2A shows that Compound A dosed at 100 mg/kg by daily oral gavage led to tumor regression in the NCI-H358 KRASG12C xenograft model, which is a model sensitive to KRASG12C inhibition alone. The spaghetti titer plot (FIG. 2B) displaying individual tumor growth is shown next to the tumor volume plot (FIG. 2A).

[0015] FIG. 3A and FIG. 3B: A compound of the present invention, Compound B, led to tumor xenograft regressions in combination with a MEK inhibitor, cobimetinib, in a model of NSCLC (KRAS G12C). FIG. 3A shows the combination of intermittent intravenous administration of Compound B at 50 mg/kg plus daily oral administration of cobimetinib at 2.5 mg/kg led to tumor regression, while each single agent led to inhibition of tumor growth. End-of-study responses were shown as waterfall plots (FIG. 3B), which indicate that 6 out of 10 mice had tumor regression in the combination group, while no tumor regression was recorded in each single agent group.

[0016] FIG. 4A and FIG. 4B: A compound of the present invention, Compound C, administered weekly with daily SHP2 inhibitor, RMC-4550, led to xenograft regressions in a model of NSCLC (KRAS G12C). In FIG. 4A, the combinatorial activity of once-weekly intravenous administration of Compound C at 60 mg/kg plus daily oral administration of SHP2 inhibitor at 30 mg/kg is shown. End-of-study responses in individual tumors were represented as a waterfall plot (FIG. 4B).

[0017] FIG. 5: A compound of the present invention, Compound D, combined with a MEK inhibitor, trametinib, durably suppressed growth in vitro in a long-term cell growth model of NSCLC (KRAS G12C).

CHEMICAL DEFINITIONS AND TERMS

[0018] In this application, unless the context otherwise requires, (i) the term “the” means “one or more”; (ii) the term “or” is used to mean “and/or” unless explicitly stated to refer only to alternatives or the alternative are mutually exclusive, although the disclosure supports a definition that refers only to alternatives and “and/or”; (iii) the terms “comprising” and “including” are understood to encompass itemized components or steps, presented by themselves or in conjunction with one or more additional components or steps; and (iv) where ranges are provided, endpoints are included.

[0019] As used herein, the term “about” is used to indicate that a value includes the standard deviation of error for the device or method to be employed to determine the value. In certain embodiments, the term “about” refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less in either direction (greater than or less than) of a stated value, unless otherwise stated or otherwise evident from the context (e.g., when such a number exceeds 100% of a possible value).

[0020] As used herein, the term “adjacent” in the context of describing adjacent atoms refers to divalent atoms that are directly linked by a covalent bond.

[0021] A “compound of the present invention” and similar terms used herein, whether explicitly noted or not, refers to Ras inhibitors described herein, including compounds of Formula I and its subformulas, and compounds of Table 1 and Table 2, as well as salts (e.g., pharmaceutically acceptable salts), solvates, hydrates, stereoisomers (including atropisomers), and tautomers thereof.

[0022] The term “wild type” refers to an entity with a structure or activity found in nature in a “normal” state or context (as contrasted with mutant, diseased, altered, etc.). Those skilled in the art will appreciate that wild type genes and polypeptides often exist in multiple different forms (e.g., alleles).

[0023] Those skilled in the art will appreciate that certain compounds described herein may exist in one or more different isomeric (e.g., stereoisomers, geometric isomers, atropisomers, tautomers) or isotopic (e.g., in which one or more atoms have been replaced by a different isotope of the atom, such as hydrogen replaced by deuterium) forms. Unless otherwise indicated or clear from the context, a depicted structure may be understood to represent any isomeric or isotopic form, individually or in combination.

[0024] The compounds described herein may be asymmetric (e.g., with one or more stereocenters). All stereoisomers, such as diastereomers and enantiomers, are intended unless otherwise indicated. Compounds of the present disclosure that contain asymmetrically substituted carbon atoms can be isolated in optically active or racemic forms. Methods for preparing optically active forms from optically active starting materials are known in the art, such as by resolution of racemic mixtures or by stereoselective synthesis. Many geometric isomers of olefins, C=N double bonds, and the like may also be present in the compounds described herein, and all such stable isomers are contemplated in the present disclosure. Cis and trans geometric isomers of the compounds of the present disclosure are described and may be isolated as a mixture of isomers or as separate isomeric forms.

[0025] In some embodiments, one or more compounds depicted herein may exist in different tautomeric forms. As will be clear from the context, unless explicitly excluded, references to such compounds encompass all such tautomeric forms. In some embodiments, tautomeric forms result from the exchange of a single bond with an adjacent double bond and the concomitant migration of a proton. In certain embodiments, a tautomeric form may be a prototropic tautomer, which is an isomeric protonation state with the same empirical formula and net charge as a reference form. Examples of moieties with prototropic tautomeric forms are ketone-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, amide-imidic acid pairs, enamine-imine pairs, and annular forms where a proton can occupy two or more positions of a heterocyclic system, such as 1H- and 3H-imidazole, 1H-, 2H-, and 4H-1,2,4-triazole, 1H- and 2H-isoindole, and 1H- and 2H-pyrazole. In some embodiments, the tautomeric forms can be in equilibrium or sterically blocked into one form by appropriate substitution. In certain embodiments, the tautomeric forms result from acetal interconversion.

[0026] Unless otherwise indicated, the structures depicted herein are also intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. Exemplary isotopes that may be incorporated into compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, such as 2H, 3H, 11C, 13C, 14C, 13N, 15N, 15O, 17O, 18O, 32P, 33P, 35S, 18F, 36Cl, 123I, and 125I. Isotopically labeled compounds (e.g., those labeled with 3H and 14C) may be useful in compound or substrate tissue distribution assays. Tritiated (i.e., 3H) and carbon-14 (i.e., 14C) isotopes may be useful for their ease of preparation and detectability. Additionally, substitution with heavier isotopes such as deuterium (i.e., 2H) may provide certain therapeutic advantages resulting from increased metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements). In some embodiments, one or more hydrogen atoms are replaced with 2H or 3H, or one or more carbon atoms are replaced with 13C- or 14C-enriched carbon. Positron-emitting isotopes such as 15O, 13N, 11C, and 18F are useful for positron emission tomography (PET) studies to examine substrate receptor occupancy. Preparations of isotopically labeled compounds are known to those of skill in the art. For example, isotopically labeled compounds can generally be prepared following procedures analogous to those disclosed for the compounds of the present invention described herein, substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

[0027] As is known in the art, many chemical entities can adopt a variety of different solid forms, such as, for example, amorphous forms or crystalline forms (e.g., polymorphs, hydrates, solvates). In some embodiments, the compounds of the present invention can be utilized in any of these forms, including in any solid form. In some embodiments, the compounds described or depicted herein can be provided or utilized in hydrate or solvate form.

[0028] At various locations throughout this specification, the substituents of the compounds of the present disclosure are disclosed in groups or in ranges. The present disclosure is specifically intended to include each and every individual subcombination of the members of these groups and ranges. For example, the term “C1-C6 alkyl” is specifically intended to disclose individually methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl, and C6 alkyl. Furthermore, where a compound includes a plurality of positions in which the substituents are disclosed in groups or in ranges, unless otherwise indicated, the present disclosure is intended to cover individual compounds and groups of compounds (e.g., genera and subgenera) containing each and every individual subcombination of members at each position.

[0029] The term “optionally substituted X” (e.g., “optionally substituted alkyl”) is intended to be equivalent to “X, wherein X is optionally substituted” (e.g., “alkyl, wherein said alkyl is optionally substituted”). It is not intended to mean that the feature “X” (e.g., alkyl) per se is optional. As described herein, certain compounds of interest may contain one or more “optionally substituted” moieties. In general, the term “substituted,” whether preceded by the term “optionally” or not, means that one or more hydrogens of the designated moiety are replaced by a suitable substituent, e.g., any of the substituents or groups described herein. Unless otherwise indicated, an “optionally substituted” group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may also be the same or different at each position. For example, in the term “optionally substituted C1-C6 alkyl-C2-C9 heteroaryl,” the alkyl moiety, the heteroaryl moiety, or both, may be optionally substituted. Combinations of substituents contemplated by the present disclosure are preferably those that result in the formation of stable or chemically viable compounds. The term “stable,” as used herein, refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and, in certain embodiments, their recovery, purification, and use for one or more of the purposes disclosed herein.

[0030] Suitable monovalent substituents on a substitutable carbon atom of an “optionally substituted” group may independently be deuterium; halogen; -(CH2)o-4R°; -(CH2)o-4OR°; -O(CH2)o- 4Ro; -O-(CH2)o-4C(O)OR°; -(CH2)O-4CH(OR°)2; -(CH2)O-4SR°; -(CH2)o-4Ph, which may be substituted by R°; -(CH2)0-4O(CH2)0-1Ph which may be substituted by R°; -CH=CHPh, which may be substituted by R°; -(CH2)0-4O(CH2)0-1-pyridyl which may be substituted by R°; saturated or unsaturated 4-8 membered heterocycloalkyl (e.g., pyridyl); saturated or unsaturated 3-8 membered cycloalkyl (e.g., cyclopropyl, cyclobutyl, or cyclopentyl); -NO2; -CN; -N3; - (CH2)0-4N(R°)2; -(CH2)0-4N(R°)C(O)R°; -N(R°)C(S)R°; -(CH2)0-4N(R°)C(O)NR°2; - N(R°)C(S)NR°2; -(CH2)0-4N(R°)C(O)OR°; - N(R°)N(R°)C(O)R°; -N(R°)N(R°)C(O)NR°2; -N(R°)N(R°)C(O)OR°; -(CH2)0-4C(O)R°; -C(S)R°; -(CH2)0- 4C(O)OR°; -(CH2)0-4-C(O)-N(Ro)2; -(CH2)0-4-C(O)-N(Ro)-S(O)2-Ro; -C(NCN)NR°2; -(CH2)0-4C(O)SR°; -(CH2)0-4C(O)OSiR°3; -(CH2)0-4OC(O)R°; -OC(O)(CH2)0-4SR°; -SC(S)SR°; -(CH2)0-4SC(O)R°; -(CH2)0-4C(O)NR°2; -C(S)NR°2; -C(S)SR°; -(CH2)0-4OC(O)NR°2; -C(O)N(OR°)R°; -C(O)C(O)R°; -C(O)CH2C(O)R°; -C(NOR°)R°;-(CH2)0-4SSR°; -(CH2)0-4S(O)2R°; -(CH2)0-4S(O)2OR°; -(CH2)0- 4OS(O)2R°; -S(O)2NR°2; -(CH2)0-4S(O)R°; -N(R°)S(O)2NR°2; -N(R°)S(O)2R°; -N(OR°)R°; -C(NOR°)NR°2; -C(NH)NR°2; -P(O)2R°; -P(O)R°2; -P(O)(OR°)2; -OP(O)R°2; -OP(O)(OR°)2; -OP(O)(OR°)R°, -SiR°3; -(C1-4 linear or branched alkylene)O-N(R°)2; or -(C1-4 linear or branched alkylene)C(O)O-N(R°)2, wherein each R° may be substituted as defined below and is independently hydrogen, -C1-6 aliphatic, -CH2Ph, -O(CH2)0-1Ph, -CH2-(5-6 membered heteroaryl ring), or a 3-6 membered saturated, partially unsaturated aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or, notwithstanding the above definition, two independent occurrences of R°, taken together with their intervening atom(s), form a 3-12 membered saturated, partially unsaturated aryl or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur which may be substituted as defined below.

[0031] Suitable monovalent substituents on R° (or the ring formed by taking two independent occurrences of R° together with their intervening atoms), may be, independently, halogen, -(CH2)O-2R*, -(haloR*), -(CH2)O-20H, -(CH2)O-20R*, -(CH2)O-2CH(OR’)2; -O(haloR*), -CN, -N3, -(CH2)O-2C(0)R*, -(CH2)O-2C(0)OH, -(CH2)O-2C(0)OR’ -(CH2)O-2SR’ -(CH2)O- 2SH, -(CH2)o-2NH2, -(CH2)o-2NHR*, -(CH2)o-2NR*2, -NO2, -SiR*3, -OSiR*3, -C(O)SR*, -(C1-4 linear or branched alkylene)C(O)OR*, or -SSR* where each R* is unsubstituted or where preceded by “halo” is substituted only by one or more halogens and is independently selected from C1-4 aliphatic, -CH2Ph, -O(CH2)o-1Ph, or a saturated, partially unsaturated, or aryl ring of 5-6 membered with o-4 heteroatoms independently selected from nitrogen, oxygen or sulfur. Suitable divalent substituents on a saturated carbon atom of R° include =O and =S.

[0032] Suitable divalent substituents on a saturated carbon atom of an “optionally substituted” group include the following: =O, =S, =NNR*2, =NNHC(O)R*, =NNHC(O)OR*, =NNHS(O)2R*, =NR*, =NOR*, -O(C(R*2))2-3O-, or -S(C(R*2))2-3S-, wherein each independent occurrence of R* is selected from hydrogen, C1-6 aliphatic which may be substituted as defined below, or a saturated, partially unsaturated 5-6 membered aryl ring unsubstituted with o-4 heteroatoms independently selected from nitrogen, oxygen or sulfur. Suitable divalent substituents that are attached to vicinal substitutable carbons of an “optionally substituted” group include: -O(CR*2)2-3O-, wherein each independent occurrence of R* is selected from hydrogen, aliphatic C1-6 may be substituted as defined below, or an unsubstituted 5-6 membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0033] Suitable substituents on the aliphatic group of R* include halogen, -R*, -(haloR*), -OH, -OR*, -O(haloR*), -CN, -C(O)OH, -C(O)OR*, - NH2, -NHR*, -NR*2, or -NO2, wherein each R* is unsubstituted or where preceded by “halo” is substituted only by one or more halogens and is independently C1-4 aliphatic, -CH2Ph, -O(CH2)0-1Ph, or a 5-6 membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0034] Suitable substituents on a substitutable nitrogen of an “optionally substituted” group include -Rt -NR^, -C(O)Rt -C(O)ORt - C(O)C(O)Rt -C(O)CH2C(O)Rt, -S(O)2Rt, -S(O)2NRt2, -C(S)NR^2, -C(NH)NR^2, or -N(Rt)S(O)2Rt; wherein each Rt is independently hydrogen, C1-6 aliphatic which may be substituted as defined below, unsubstituted -OPh, or an unsubstituted 3-6 membered saturated, partially unsaturated aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or, notwithstanding the above definition, two independent occurrences of Rt, taken together with their intervening atom(s) form a saturated, partially unsaturated or unsubstituted 3-12 membered aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0035] Suitable substituents on an aliphatic group of Rt are independently halogen, -R*, -(haloR*), -OH, -OR*, -O(haloR*), -CN, -C(O)OH, -C(O)OR*, -NH2, -NHR*, -NR*2, or -NO2, wherein each R* is unsubstituted or where preceded by "halo" is substituted only by one or more halogens and is independently C1-4 aliphatic, -CH2Ph, -O(CH2)0-1Ph, or a 5-6 membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents on a saturated carbon atom of Rt include =O and =S.

[0036] The term “acetyl”, as used in this document, refers to the group -C(O)CH3.

[0037] The term “alkoxy”, as used in this document, refers to an O-C1-C20 alkyl group, in which the alkoxy group is bonded to the remainder of the compound through an oxygen atom.

[0038] The term “alkyl,” as used herein, refers to a linear or branched, saturated monovalent hydrocarbon group containing from 1 to 20 (e.g., from 1 to 10 or from 1 to 6) carbons. In some embodiments, an alkyl group is unbranched (i.e., linear); in some embodiments, an alkyl group is branched. Alkyl groups are exemplified by, but not limited to, methyl, ethyl, n- and iso-propyl, n-, sec-, iso- and tert-butyl, and neopentyl.

[0039] The term “alkylene” as used herein represents a saturated divalent hydrocarbon group derived from a straight or branched chain saturated hydrocarbon by the removal of two hydrogen atoms and is exemplified by methylene, ethylene, isopropylene, and the like. The term “Cx-Cy alkylene” represents alkylene groups having between x and y carbons. Exemplary values for x are 1, 2, 3, 4, 5, and 6, and exemplary values for y are 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, or 20 (e.g., C1-C6, C1-C10, C2-C20, C2-C6, C2-C10, or C2-C20 alkylene). In some embodiments, the alkylene may be further substituted with 1, 2, 3, or 4 substituent groups as defined herein.

[0040] The term “alkenyl” as used herein represents monovalent straight or branched chain groups of, unless otherwise specified, 2 to 20 carbons (e.g., 2 to 6 or 2 to 10 carbons) containing one or more carbon-carbon double bonds and is exemplified by ethenyl, 1-propenyl, 2-propenyl, 2-methyl-1-propenyl, 1-butenyl, and 2-butenyl. Alkenyls include cis and trans isomers. The term “alkenylene” as used herein represents divalent straight or branched chain groups of, unless otherwise specified, 2 to 20 carbons (e.g., 2 to 6 or 2 to 10 carbons) containing one or more carbon-carbon double bonds.

[0041] The term “alkynyl” as used herein represents monovalent straight or branched chain groups of 2 to 20 carbon atoms (e.g., 2 to 4, 2 to 6, or 2 to 10 carbons) containing a carbon-carbon triple bond and is exemplified by ethynyl and 1-propynyl.

[0042] The term “alkynyl sulfone”, as used herein, represents a group comprising the structure , where R is any chemically viable substituent described herein.

[0043] The term “amino”, as used herein, represents -N(R^)2, for example, -NH2 and -N(CH3)2.

[0044] The term “aminoalkyl,” as used herein, represents an alkyl moiety substituted at one or more carbon atoms with one or more amino moieties.

[0045] The term “amino acid,” as used herein, refers to a molecule having a side chain, an amino group, and an acidic group (e.g., -CO2H or -SO3H), wherein the amino acid is linked to the parent molecular group by the side chain, amino group, or acidic group (e.g., the side chain). As used herein, the term “amino acid,” in its broadest sense, refers to any compound or substance that can be incorporated into a polypeptide chain, for example, through the formation of one or more peptide bonds. In some embodiments, an amino acid has the general structure H2N-C(H)(R)-COOH. In some embodiments, an amino acid is a naturally occurring amino acid. In some embodiments, an amino acid is a synthetic amino acid; in some embodiments, an amino acid is a D-amino acid; in some embodiments, an amino acid is an L-amino acid. “Standard amino acid” refers to any of the twenty standard L-amino acids commonly found in naturally occurring peptides. Examples of amino acids include alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, optionally substituted hydroxylnorvaline, isoleucine, leucine, lysine, methionine, norvaline, ornithine, phenylalanine, proline, pyrrolysine, selenocysteine, serine, taurine, threonine, tryptophan, tyrosine, and valine.

[0046] The term “aryl” as used herein represents a monovalent monocyclic, bicyclic or multicyclic ring system formed by carbon atoms, in which the ring attached to the pendant group is aromatic. Examples of aryl groups are phenyl, naphthyl, phenanthrenyl and anthracenyl. An aryl ring may be attached to its pendant group at any heteroatom or carbon ring atom that results in a stable structure and any of the ring atoms may be optionally substituted unless otherwise specified.

[0047] The term “C0” as used herein represents a bond. For example, part of the term -N(C(O)-(C0-C5 alkylene-H)- includes - N(C(O)-(C0 alkylene-H)-, which is also represented by -N(C(O)-H)-.

[0048] The terms “carbocyclic” and “carbocyclyl,” as used herein, refer to a monovalent, optionally substituted C3-C12 monocyclic, bicyclic, or tricyclic ring structure, which may be bridged, fused, or spirocyclic, in which all rings are formed by carbon atoms and at least one ring is non-aromatic. Carbocyclic structures include cycloalkyl, cycloalkenyl, and cycloalkynyl groups. Examples of carbocyclyl groups are cyclohexyl, cyclohexenyl, cyclooctynyl, 1,2-dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, fluorenyl, indenyl, indanyl, decalinyl, and the like. A carbocyclic ring may be attached to its pendant group at any ring atom that results in a stable structure, and any of the ring atoms may be optionally substituted unless otherwise specified.

[0049] The term “carbonyl”, as used in this document, represents a C(O) group, which can also be represented as C=O.

[0050] The term “carboxyl” as used herein means -CO2H, (C=O)(OH), COOH or C(O)OH or the unprotonated counterparts.

[0051] The term “cyano”, as used in this document, represents a -CN group.

[0052] The term “cycloalkyl” as used herein represents a monovalent saturated cyclic hydrocarbon group, which may be bridged, fused or spirocyclic having from three to eight carbons in the ring, unless otherwise specified, and is exemplified by cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cycloheptyl.

[0053] The term “cycloalkenyl” as used herein represents a monovalent, non-aromatic, saturated cyclic hydrocarbon group, which may be bridged, fused or spirocyclic having from three to eight carbons in the ring, unless otherwise specified, and containing one or more carbon-carbon double bonds.

[0054] The term “diastereomer” as used herein means stereoisomers that are not mirror images of each other and are not superimposable on each other.

[0055] The term “enantiomer” as used herein means each individual optically active form of a compound of the invention having an optical purity or enantiomeric excess (as determined by standard methods in the art) of at least 80% (i.e. at least 90% of one enantiomer and at most 10% of the other enantiomer), preferably at least 90% and more preferably at least 98%.

[0056] The term “guanidinyl” refers to a group with the structure: where each R is, independently, any chemically viable substituent described herein.

[0057] The term “guanidinoalkyl alkyl”, as used herein, represents an alkyl moiety substituted on one or more carbon atoms by one or more guanidinyl moieties.

[0058] The term “haloacetyl”, as used in this document, refers to an acetyl group in which at least one of the hydrogens has been replaced by a halogen.

[0059] The term “haloalkyl”, as used herein, represents an alkyl moiety substituted on one or more carbon atoms with one or more of the same from different halogen moieties.

[0060] The term “halogen” as used herein represents a halogen selected from bromine, chlorine, iodine or fluorine.

[0061] The term “heteroalkyl,” as used herein, refers to an “alkyl” group, as defined herein, in which at least one carbon atom has been replaced by a heteroatom (e.g., an O, N, or S atom). The heteroatom may appear in the middle or at the end of the radical.

[0062] The term “heteroaryl,” as used herein, represents a monovalent, monocyclic or polycyclic ring structure that contains at least one fully aromatic ring: that is, they contain 4n+2 pi electrons within the monocyclic or polycyclic ring system and contain at least one ring heteroatom selected from N, O or S in that aromatic ring. Exemplary unsubstituted heteroaryl groups are 1 to 12 (e.g., 1 to 11, 1 to 10, 1 to 9, 2 to 12, 2 to 11, 2 to 10 or 2 to 9) carbons. The term “heteroaryl” includes bicyclic, tricyclic and tetracyclic groups in which any of the above heteroaromatic rings is fused to one or more aryl or carbocyclic rings, for example, a phenyl ring or a cyclohexane ring. Examples of heteroaryl groups include, but are not limited to, pyridyl, pyrazolyl, benzooxazolyl, benzoimidazolyl, benzothiazolyl, imidazolyl, thiazolyl, quinolinyl, tetrahydroquinolinyl, and 4-azaindolyl. A heteroaryl ring may be attached to its pendant group at any ring atom that results in a stable structure, and any of the ring atoms may be optionally substituted unless otherwise specified. In some embodiments, the heteroaryl is substituted by 1, 2, 3, or 4 substituent groups.

[0063] The term “heterocycloalkyl” as used herein represents a monovalent monocyclic, bicyclic or polycyclic ring system, which may be bridged, fused or spirocyclic, in which at least one ring is non-aromatic and in which the non-aromatic ring contains one, two, three or four heteroatoms independently selected from the group consisting of nitrogen, oxygen and sulfur. The 5-membered ring has zero to two double bonds, and the 6- and 7-membered rings have zero to three double bonds. Exemplary unsubstituted heterocycloalkyl groups are 1 to 12 (e.g., 1 to 11, 1 to 10, 1 to 9, 2 to 12, 2 to 11, 2 to 10 or 2 to 9) carbons. The term "heterocycloalkyl" also represents a heterocyclic compound having a bridged multicyclic structure in which one or more carbons or heteroatoms bridge two non-adjacent members of a monocyclic ring, e.g., a quinuclidinyl group. The term "heterocycloalkyl" includes bicyclic, tricyclic, and tetracyclic groups in which any of the above heterocyclic rings is fused to one or more aromatic, carbocyclic, heteroaromatic, or heterocyclic rings, e.g., an aryl ring, a cyclohexane ring, a cyclohexene ring, a cyclopentane ring, a cyclopentene ring, a pyridine ring, or a pyrrolidine ring. Examples of heterocycloalkyl groups are pyrrolidinyl, piperidinyl, 1,2,3,4-tetrahydroquinolinyl, decahydroquinolinyl, dihydropyrrolopyridine, and decahydronaphthyridinyl. A heterocycloalkyl ring may be attached to its pendant group at any ring atom that results in a stable structure, and any of the ring atoms may be optionally substituted unless otherwise specified.

[0064] The term “hydroxy” as used herein represents an -OH group.

[0065] The term “hydroxyalkyl,” as used herein, represents an alkyl moiety substituted on one or more carbon atoms with one or more -OH moieties.

[0066] The term “isomer” as used herein means any tautomer, stereoisomer, atropiomer, enantiomer or diastereomer of any compound of the invention. It is recognized that the compounds of the invention may have one or more chiral centers or double bonds and therefore exist as stereoisomers, such as double bond isomers (i.e., geometric E/Z isomers) or diastereomers (e.g., enantiomers (i.e., (+) or (-)) or cis/trans isomers). According to the invention, the chemical structures depicted herein, and therefore the compounds of the invention, encompass all corresponding stereoisomers, i.e., both the stereomerically pure form (e.g., geometrically pure, enantiomerically pure or diastereomerically pure) and enantiomeric and stereoisomeric mixtures, e.g., racemates. Enantiomeric and stereoisomeric mixtures of compounds of the invention can typically be resolved into their component enantiomers or stereoisomers by well-known methods, such as chiral phase gas chromatography, chiral phase high-performance liquid chromatography, crystallizing the compound as a chiral salt complex, or crystallizing the compound in a chiral solvent. Enantiomers and stereoisomers can also be obtained from stereomerically or enantiomerically pure intermediates, reagents, and catalysts by well-known asymmetric synthetic methods.

[0067] As used herein, the term “linker” refers to a divalent organic moiety that connects the B moiety to the W moiety in a compound of Formula I, such that the resulting compound is capable of achieving an IC50 of 2 uM or less in the Ras-RAF disruption assay protocol provided in the Examples below and provided herein.

[0068] The objective of this biochemical assay is to measure the ability of test compounds to facilitate the formation of the ternary complex between a nucleotide-loaded RAS isoform and cyclophilin A; the resulting ternary complex disrupts binding to a BRAFRBD construct, inhibiting RAS signaling through a RAF effector.

[0069] In assay buffer containing 25 mM HEPES pH 7.3, 0.002% Tween20, 0.1% BSA, 100 mM NaCl 5 mM MgCl2, untagged Cyclophilin A, His6-K-Ras-GMPPNP (or other Ras variant), and GST-BRAFRBD are combined in a 384-well assay plate at final concentrations of 25 μM, 12.5 nM, and 50 nM, respectively. Compound is present in plate wells as a 10-point, 3-fold dilution series starting at a final concentration of 30 μM. After incubation at 25°C for 3 hours, a mixture of Anti-His Eu-W1024 and anti-GST allophycocyanin is then added to the assay sample wells at final concentrations of 10 nM and 50 nM, respectively, and the reaction incubated for an additional 1.5 hours. The TR-FRET signal is read on a microplate reader (Ex 320 nm, Em 665/615 nm). Compounds that facilitate disruption of a Ras:RAF complex are identified as those that elicit a decrease in the TR-FRET ratio relative to DMSO control wells.

[0070] In some embodiments, the linker comprises 20 or fewer linear atoms. In some embodiments, the linker comprises 15 or fewer linear atoms. In some embodiments, the linker comprises 10 or fewer linear atoms. In some embodiments, the linker has a molecular weight of less than 500 g/mol. In some embodiments, the linker has a molecular weight of less than 400 g/mol. In some embodiments, the linker has a molecular weight of less than 300 g/mol. In some embodiments, the linker has a molecular weight of less than 200 g/mol. In some embodiments, the linker has a molecular weight of less than 100 g/mol. In some embodiments, the linker has a molecular weight of less than 50 g/mol.

[0071] As used herein, a “monovalent organic fraction” is less than 500 kDa. In some embodiments, a “monovalent organic fraction” is less than 400 kDa. In some embodiments, a “monovalent organic fraction” is less than 300 kDa. In some embodiments, a “monovalent organic fraction” is less than 200 kDa. In some embodiments, a “monovalent organic fraction” is less than 100 kDa. In some embodiments, a “monovalent organic fraction” is less than 50 kDa. In some embodiments, a “monovalent organic fraction” is less than 25 kDa. In some embodiments, a “monovalent organic fraction” is less than 20 kDa. In some embodiments, a “monovalent organic fraction” is less than 15 kDa. In some embodiments, a “monovalent organic fraction” is less than 10 kDa. In some embodiments, a “monovalent organic moiety” is less than 1 kDa. In some embodiments, a “monovalent organic moiety” is less than 500 g/mol. In some embodiments, a “monovalent organic moiety” ranges from 500 g/mol to 500 kDa.

[0072] The term “stereoisomer”, as used herein, refers to all possible different isomeric and conformational forms that a compound may possess (e.g., a compound of any formula described herein), in particular all possible stereochemically and conformationally isomeric forms, all diastereomers, enantiomers or conformers of the basic molecular structure, including atropisomers. Some compounds of the present invention may exist in different tautomeric forms, all the latter being included within the scope of the present invention.

[0073] The term “sulfonyl”, as used in this document, represents a -S(O)2- group.

[0074] The term “thiocarbonyl”, as used herein, refers to a -C(S)- group.

[0075] The term “vinyl ketone,” as used herein, refers to a group comprising a carbonyl group directly attached to a carbon-carbon double bond.

[0076] The term “vinyl sulfone,” as used herein, refers to a group comprising a directed sulfonyl group attached to a carbon-carbon double bond.

[0077] The term “inone”, as used in this document, refers to a group comprising the structure , where R is any chemically viable substituent described herein.

[0078] Those skilled in the art reading the present disclosure will appreciate that certain compounds described herein may be provided or utilized in any of a variety of forms, such as, for example, salt forms, protected forms, prodrug forms, ester forms, isomeric forms (e.g., optical or structural isomers), isotopic forms, etc. In some embodiments, reference to a particular compound may refer to a specific form of that compound. In some embodiments, reference to a particular compound may refer to that compound in any form. In some embodiments, for example, a preparation of a single stereoisomer of a compound may be considered a different form of the compound than a racemic mixture of the compound; a particular salt of a compound may be considered a different form than another salt form of the compound; a preparation containing one conformational isomer ((Z) or (E)) of a double bond may be considered a different form from one containing the other conformational isomer ((E) or (Z)) of the double bond; a preparation in which one or more atoms is a different isotope than that present in a reference preparation may be considered a different form.

DETAILED DESCRIPTION COMPOUNDS

[0079] Provided herein are inhibitors of Ras. The approach described herein involves the formation of a high-affinity three-component complex, or conjugate, between a synthetic ligand and two intracellular proteins that do not interact under normal physiological conditions: the target protein of interest (e.g., Ras) and a widely expressed cytosolic chaperone (presenting protein) in the cell (e.g., cyclophilin A). More specifically, in some embodiments, the Ras inhibitors described herein induce a novel binding pocket in Ras, leading to the formation of a high-affinity triple complex, or conjugate, between the Ras protein and the widely expressed cytosolic chaperone, cyclophilin A (CYPA). Without being bound by theory, the inventors believe that one way in which the inhibitory effect on Ras is effected by the compounds of the invention and the complexes, or conjugates, they form is by steric occlusion of the interaction site between Ras and downstream effector molecules, such as RAF, that are necessary to propagate the oncogenic signal.

[0080] Without being limited by theory, the inventors postulate that covalent and non-covalent interactions of a compound of the present invention with Ras and chaperone protein (e.g., cyclophilin A) may contribute to the inhibition of Ras activity. In some embodiments, a compound of the present invention forms a covalent adduct with a side chain of a Ras protein (e.g., a sulfhydryl side chain of the cysteine at position 12 or 13 of a mutant Ras protein). Covalent adducts may also be formed with other Ras side chains. Additionally, or alternatively, non-covalent interactions may be at play: for example, van der Waals, hydrophobic, hydrophilic, and hydrogen bonding interactions, and combinations thereof, may contribute to the ability of compounds of the present invention to form complexes and act as Ras inhibitors. Accordingly, a variety of Ras proteins can be inhibited by compounds of the present invention (e.g., K-Ras, N-Ras, H-Ras, and mutants thereof at positions 12, 13, and 61, such as G12C, G12D, G12V, G12S, G13C, G13D, and Q61L, and others described herein).

[0081] Methods for determining covalent adduct formation are known in the art. One method for determining covalent adduct formation is to perform a “crosslinking” assay, such as under these conditions (Note - the following protocol describes a procedure for monitoring the crosslinking of K-Ras G12C (GMP-PNP) to a compound of the invention. This protocol may also be performed substituting other RAS proteins or nucleotides).

[0082] The objective of this biochemical assay is to measure the ability of test compounds to covalently label nucleotide-loaded K-Ras isoforms. In assay buffer containing 12.5 mM HEPES pH 7.4, 75 mM NaCl, 1 mM MgCl2, 1 mM BME, 5 μM Cyclophilin A, and 2 μM test compound, a 5 μM stock of K-Ras loaded with GMP-PNP(1-169)G12C is diluted 10-fold to yield a final concentration of 0.5 μM; with final sample volume being 100 μL.

[0083] The sample is incubated at 25°C for up to 24 hours before quenching by the addition of 10 μL of 5% formic acid. The quenched samples are centrifuged at 15,000 rpm for 15 minutes in a benchtop centrifuge before injecting a 10 μL aliquot onto a C4 reversed phase column and eluting into the mass spectrometer with an increasing gradient of acetonitrile in the mobile phase. Raw data analysis can be performed using Waters MassLynx MS software, with % binding calculated from the deconvoluted protein peaks for both labeled and unlabeled K-Ras.

[0084] Accordingly, provided herein is a compound, or pharmaceutically acceptable salt thereof, having the structure of Formula I: Formula Iwhere the dotted lines represent zero, one, two, three, or four nonadjacent double bonds;A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 10-membered heteroarylene;B is absent, is -CH(R9)-, >C=CR9R9', or >CR9R9'where the carbon is bonded to the carbonyl carbon of -N(R11)C(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;G is optionally substituted C1-C4 alkylene, Optionally substituted C1-C4 alkenylene, optionally substituted C1-C4 heteroalkylene, -C(O)O-CH(R6)- where C is attached to -C(R7R8)-, -C(O)NH-CH(R6)- where C is attached to -C(R7R8)-, optionally substituted C1-C4 heteroalkylene, or 3- to 8-membered heteroarylene;L is absent or is a ligand;W is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone, a haloacetal, or an alkynyl sulfone;X1 is optionally substituted C1-C2 alkylene, NR, O, or S(O)n;X2 is O or NH;X3 is N or CH;n is 0, 1, or 2;R is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2C4 alkenyl, C2-C4 optionally substituted alkynyl, C(O)R', C(O)OR', C(O)N(R')2, S(O)R', S(O)2R', or S(O)2N(R')2; each R' is independently H or optionally substituted C1-C4 alkyl; Y1 is C, CH, or N; Y2, Y3, Y4, and Y7 are independently C or N; Y 5 is CH, CH 2 , or N; Y 6 is C(O), CH, CH 2 , or N; R 1 is cyano, optionally substituted C 1-C 6 alkyl, optionally substituted C 1-C 6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl, or R 1 and R 2 combine with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl; R 2 is absent, is hydrogen, optionally substituted C 1-C 6 alkyl, optionally substituted C 2-C 6 alkenyl, optionally substituted C 2-C 6 alkynyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl; R3 is absent, or R2 and R3 combine with the atom to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or optionally substituted 3- to 14-membered heterocycloalkyl; R4 is absent, is hydrogen, halogen, cyano, or methyl optionally substituted with 1 to 3 halogens; R5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl, or cyclobutyl; R6 is hydrogen or methyl; R7 is hydrogen, halogen, or optionally substituted C1-C3 alkyl, or R6 and R7 combine with the carbon atoms to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7 and R8 combine with the carbon atom to which they are attached to form C=CR7'R8'; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;R7a and R8a are independently hydrogen, halo, C1-C3 alkyl, or combine with the carbon to which they are attached to form a carbonyl;R7' is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R8' is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7' and R8' combine with the carbon atom to which they are attached to form optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R9 is H, F optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl, or R9 and L combine with the atoms to which they are attached to form form an optionally substituted 3- to 14-membered heterocycloalkyl; R9' is hydrogen or optionally substituted C1-C6 alkyl; or R9 and R9', combined with the atoms to which they are attached, form a 3- to 6-membered cycloalkyl or a 3- to 6-membered heterocycloalkyl; R10 is hydrogen, halo, hydroxy, C1-C3 alkoxy, or C1-C3 alkyl; R10a is hydrogen or halo; R11 is hydrogen or C1-C3 alkyl; and R21 is hydrogen or C1-C3 alkyl (e.g., methyl).

[0085] In some embodiments, R9 is H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl.

[0086] In some embodiments, R21 is hydrogen.

[0087] In some embodiments, provided herein is a compound, or pharmaceutically acceptable salt thereof, having the structure of Formula Ia: Formula Iawhere the dotted lines represent zero, one, two, three, or four nonadjacent double bonds;A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 10-membered heteroarylene;B is -CH(R9)- or >C=CR9R9' where the carbon is bonded to the carbonyl carbon of -N(R11)C(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;G is optionally substituted C1-C4 alkylene, C1-C4 alkenylene optionally substituted, optionally substituted C1-C4 heteroalkylene, -C(O)O-CH(R6)- where C is attached to -C(R7R8)-, -C(O)NH-CH(R6)- where C is attached to -C(R7R8)-, optionally substituted C1-C4 heteroalkylene, or 3- to 8-membered heteroarylene;L is absent or is a ligand;W is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone, a haloacetal, or an alkynyl sulfone;X1 is optionally substituted C1-C2 alkylene, NR, O, or S(O)n;X2 is O or NH;X3 is N or CH;n is 0, 1, or 2;R is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, C(O)R', C(O)OR', C(O)N(R')2, S(O)R', S(O)2R', or S(O)2N(R')2; each R' is independently H or optionally substituted C1-C4 alkyl; Y1 is C, CH, or N; Y2, Y3, Y4, and Y7 are independently C or N; Y 5 is CH, CH 2 , or N; Y 6 is C(O), CH, CH 2 , or N; R 1 is cyano, optionally substituted C 1-C 6 alkyl, optionally substituted C 1-C 6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl, or R 1 and R 2 combine with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl; R 2 is absent, is hydrogen, optionally substituted C 1-C 6 alkyl, optionally substituted C 2-C 6 alkenyl, optionally substituted C 2-C 6 alkynyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl; R3 is absent, or R2 and R3 combine with the atom to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or optionally substituted 3- to 14-membered heterocycloalkyl; R4 is absent, is hydrogen, halogen, cyano, or methyl optionally substituted by 1 to 3 halogens; R5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl, or cyclobutyl; R6 is hydrogen or methyl; R7 is hydrogen, halogen, or optionally substituted C1-C3 alkyl, or R6 and R7 combine with the carbon atoms to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7 and R8 combine with the carbon atom to which they are attached to form C=CR7'R8'; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;R7a and R8a are independently hydrogen, halo, C1-C3 alkyl, or combine with the carbon to which they are attached to form a carbonyl;R7' is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R8' is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7' and R8' combine with the carbon atom to which they are attached to form optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl, or R9 and L combine with the atoms to which they are attached to form a optionally substituted 3- to 14-membered heterocycloalkyl;R9' is hydrogen or optionally substituted C1-C6 alkyl;R10 is hydrogen, halo, hydroxy, C1-C3 alkoxy, or C1-C3 alkyl;R10a is hydrogen or halo; andR11 is hydrogen or C1-C3 alkyl.

[0088] In some embodiments, the disclosure features a compound, or pharmaceutically acceptable salt thereof, of structural Formula Ib: Formula Iwhere the dotted lines represent zero, one, two, three, or four nonadjacent double bonds;A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 6-membered heteroarylene;B is -CH(R9)- where the carbon is bonded to the carbonyl carbon of -N(R11)C(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;G is optionally substituted C1-C4 alkylene, optionally substituted C1-C4 alkenylene, C1-C4 optionally substituted heteroalkylene, - C(O)O-CH(R6)- where C is attached to -C(R7R8)-, -C(O)NH-CH(R6)- where C is attached to -C(R7R8)-, optionally substituted C1-C4 heteroalkylene, or 3- to 8-membered heteroarylene;L is absent or is a ligand;W is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone, a haloacetal, or an alkynyl sulfone;X1 is optionally substituted C1-C2 alkylene, NR, O, or S(O)n;X2 is O or NH;X3 is N or CH;n is 0, 1, or 2;R is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, C(O)R', C(O)OR', C(O)N(R')2, S(O)R', S(O)2R', or S(O)2N(R')2; each R' is independently H or optionally substituted C1-C4 alkyl; Y1 is C, CH, or N; Y2, Y3, Y4, and Y7 are independently C or N; Y5 and Y6 are independently CH or N; R1 is cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl; R2 is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl; R3 is absent, or R2 and R3 combine with the atom to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or optionally substituted 3- to 14-membered heterocycloalkyl; R4 is absent, is hydrogen, halogen, cyano, or methyl optionally substituted by 1 to 3 halogens; R5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl, or cyclobutyl; R6 is hydrogen or methyl; R7 is hydrogen, halogen, or optionally substituted C1-C3 alkyl, or R6 and R7 combine with the carbon atoms to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7 and R8 combine with the carbon atom to which they are attached to form C=CR7'R8'; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;R7' is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R8' is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7' and R8' combine with the carbon atom to which they are attached to form optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; R10 is hydrogen, hydroxy, C1-C3 alkoxy, or C1-C3 alkyl; and R11 is hydrogen or C1-C3 alkyl.

[0089] In some embodiments of the compounds of the present invention, G is optionally substituted C1-C4 heteroalkylene.

[0090] In some embodiments, a compound having the structure of Formula Ic, or a pharmaceutically acceptable salt thereof, is provided: Formula Iwhere the dotted lines represent zero, one, two, three, or four nonadjacent double bonds;A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 6-membered heteroarylene;B is -CH(R9)- where the carbon is bonded to the carbonyl carbon of -N(R11)C(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;L is absent or is a ligand;W is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone, or a alkynyl sulfone;X2 is O or NH;X3 is N or CH;n is 0, 1, or 2;R is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, C(O)R', C(O)OR', C(O)N(R')2, S(O)R', S(O)2R', or S(O)2N(R')2;each R' is independently H or optionally substituted C1-C4 alkyl;Y1 is C, CH, or N;Y2, Y3, Y4, and Y7 are independently C or N;Y5 and Y6 are independently CH or N;R1 is cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, 3- to optionally substituted 6-membered, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl or optionally substituted 5- to 10-membered heteroaryl;R2 is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl; R3 is absent, or R2 and R3 combine with the atom to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or optionally substituted 3- to 14-membered heterocycloalkyl; R4 is absent, is hydrogen, halogen, cyano, or methyl optionally substituted by 1 to 3 halogens; R5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl, or cyclobutyl; R6 is hydrogen or methyl; R7 is hydrogen, halogen, or optionally substituted C1-C3 alkyl, or R6 and R7 combine with the carbon atoms to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7 and R8 combine with the carbon atom to which they are attached to form C=CR7'R8'; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;R7' is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R8' is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7' and R8' combine with the carbon atom to which they are attached to form optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; R10 is hydrogen, hydroxy, C1-C3 alkoxy, or C1-C3 alkyl; and R11 is hydrogen or C1-C3 alkyl.

[0091] In some embodiments of the compounds of the present invention, X2 is NH. In some embodiments, X3 is CH. In some embodiments, R11 is hydrogen. In some embodiments, R11 is C1-C3 alkyl. In some embodiments, R11 is methyl.

[0092] In some embodiments, a compound of the present invention has the structure of Formula Id, or a pharmaceutically acceptable salt thereof: Formula Same where the dotted lines represent zero, one, two, three, or four nonadjacent double bonds;A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 6-membered heteroarylene;B is -CH(R9)- where the carbon is bonded to the carbonyl carbon of NHC(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene; L is absent or is a ligand; W is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone, or an alkynyl sulfone; n is 0, 1, or 2; R is hydrogen, cyano, optionally substituted C 1-C 4 alkyl, optionally substituted C 2-C 4 alkenyl, optionally substituted C 2-C 4 alkynyl, C(O)R', C(O)OR', C(O)N(R')2, S(O)R', S(O)2R', or S(O)2N(R')2; each R' is independently H or optionally substituted C 1-C 4 alkyl; Y 1 is C, CH, or N; Y 2 , Y 3 , Y 4, and Y 7 are independently C or N; Y 5 and Y 6 are independently CH or N; R 1 is cyano, optionally substituted C 1-C 6 alkyl, optionally substituted C 1-C 6 heteroalkyl substituted, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl; R2 is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl; R3 is absent, or R2 and R3 combine with the atom to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or optionally substituted 3- to 14-membered heterocycloalkyl; R4 is absent, is hydrogen, halogen, cyano, or methyl optionally substituted with 1 to 3 halogens; R5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl or cyclobutyl;R6 is hydrogen or methyl; R7 is hydrogen, halogen, or optionally substituted C1-C3 alkyl, or R6 and R7 combine with the carbon atoms to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7 and R8 combine with the carbon atom to which they are attached to form C=CR7'R8'; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;R7' is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R8' is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7' and R8' combine with the carbon atom to which they are attached to form optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; eR10 is hydrogen, hydroxy, C1-C3 alkoxy, or C1-C3 alkyl.

[0093] In some embodiments of a compound of the present invention, X1 is optionally substituted C1-C2 alkylene. In some embodiments, X1 is methylene. In some embodiments, X1 is methylene substituted with a C1-C6 alkyl group or a halogen. In some embodiments, X1 is -CH(Br)-. In some embodiments, X1 is -CH(CH3)-. In some embodiments, R5 is hydrogen. In some embodiments, R5 is C1-C4 alkyl optionally substituted with halogen. In some embodiments, R5 is methyl. In some embodiments, Y4 is C. In some embodiments, R4 is hydrogen. In some embodiments, Y5 is CH.

[0094] In some embodiments, Y6 is CH. In some embodiments, Y1 is C. In some embodiments, Y2 is C. In some embodiments, Y3 is N. In some embodiments, R3 is absent. In some embodiments, Y7 is C.

[0095] In some embodiments, a compound of the present invention has the structure of Formula Ie, or a pharmaceutically acceptable salt thereof: Formula Ie where A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 6-membered heteroarylene; B is -CH(R9)- where the carbon is bonded to the carbonyl carbon of NHC(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene; L is absent or is a ligand; W is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone, or an alkynyl sulfone; R1 is cyano, optionally substituted C1-C6 alkyl, Optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl or optionally substituted 5- to 10-membered heteroaryl;R2 is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl; R3 is absent, or R2 and R3 combine with the atom to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or optionally substituted 3- to 14-membered heterocycloalkyl; R5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl or cyclobutyl; R6 is hydrogen or methyl; R7 is hydrogen, halogen, or optionally substituted C1-C3 alkyl, or R6 and R7 combine with the carbon atoms to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7 and R8 combine with the carbon atom to which they are attached to form C=CR7'R8'; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;R7' is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R8' is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7' and R8' combine with the carbon atom to which they are attached to form optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; and R10 is hydrogen, hydroxy, C1-C3 alkoxy, or C1-C3 alkyl.

[0096] In some embodiments of a compound of the present invention, R6 is hydrogen. In some embodiments, R2 is hydrogen, cyano, optionally substituted C1C6 alkyl, optionally substituted 3-6 membered cycloalkyl, or optionally substituted 3-6 membered heterocycloalkyl. In some embodiments, R2 is optionally substituted C1-C6 alkyl. In some embodiments, R2 is fluoroalkyl. In embodiments, R2 is ethyl. In some embodiments, R2 is -CH2CF3. In some embodiments, R2 is C2-C6 alkynyl. In some embodiments, R2 is -CHC=CH. In some embodiments, R2 is -CH2C=CCH3. In some embodiments, R7 is optionally substituted C1-C3 alkyl. In some embodiments, R7 is C1-C3 alkyl. In some embodiments, R8 is optionally substituted C1-C3 alkyl. In some embodiments, R8 is C1-C3 alkyl.

[0097] In some embodiments, a compound of the present invention has the structure of Formula If, or a pharmaceutically acceptable salt thereof: Formula Ifem where A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 6-membered heteroarylene;B is -CH(R9)- where the carbon is bonded to the carbonyl carbon of NHC(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;L is absent or is a ligand;W is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone, or an alkynyl sulfone;R1 is cyano, optionally substituted C1-C6 alkyl, C1-C6 optionally substituted heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl; R2 is C1-C6 alkyl or 3- to 6-membered cycloalkyl; R7 is C1-C3 alkyl; R8 is C1-C3 alkyl; and R9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl.

[0098] In some embodiments of a compound of the present invention, R1 is optionally substituted 6- to 10-membered aryl, optionally substituted 3- to 6-membered cycloalkenyl, or optionally substituted 5- to 10-membered heteroaryl. In some embodiments, R1 is optionally substituted 6-membered aryl, optionally substituted 6-membered cycloalkenyl, or optionally substituted 6-membered heteroaryl.

[0099] In some embodiments of a compound of the present invention, R1 is or , or a stereoisomer (e.g., atropisomer) thereof. In some embodiments of a compound of the present invention, R 1 is , or a stereoisomer (e.g., atropisomer) thereof. In some embodiments of a compound of the present invention, R 1 is

[00100] In some embodiments, a compound of the present invention has the structure of Formula Ig, or a pharmaceutically acceptable salt thereof: Formula Igem where A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 6-membered heteroarylene; B is -CH(R9)- where the carbon is bonded to the carbonyl carbon of NHC(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene; L is absent or is a ligand; W is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone, or an alkynyl sulfone; R2 is C1-C6 alkyl, C1-C6 fluoroalkyl, or 3- to 6-membered cycloalkyl;R7 is C1-C3 alkyl;R8 is C1-C3 alkyl; andR9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;Xe and Xf are independently N or CH; and R12 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, or optionally substituted 3- to 6-membered heterocycloalkylene.

[00101] In some embodiments of a compound of the present invention, Xe is N and Xf is CH. In some embodiments, Xe is CH and Xf is N.

[00102] In some embodiments of a compound of the present invention, R12 is optionally substituted C1-C6 heteroalkyl. In some embodiments, R12 is . In some embodiments, R12 is

[00103] In some embodiments, a compound of the present invention has the structure of Formula VI, or a pharmaceutically acceptable salt thereof: Formula VIwhere the dotted lines represent zero, one, two, three, or four nonadjacent double bonds;A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene (e.g., phenyl or phenol), or optionally substituted 5- to 10-membered heteroarylene;B is absent, -CH(R9)-, >C=CR9R9', or >CR9R9' where the carbon is bonded to the carbonyl carbon of -N(R11)C(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;G is Optionally substituted C1-C4 alkylene, optionally substituted C1-C4 alkenylene, optionally substituted C1-C4 heteroalkylene, -C(O)O-CH(R6)- where C is attached to -C(R7R8)-, -C(O)NH-CH(R6)- where C is attached to -C(R7R8)-, optionally substituted C1-C4 heteroalkylene, or 3- to 8-membered heteroarylene;L is absent or is a ligand;W is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone, a haloacetal, or an alkynyl sulfone;X1 is optionally substituted C1-C2 alkylene, NR, O, or S(O)n;X2 is O or NH;X3 is N or CH;n is 0, 1, or 2;R is hydrogen, cyano, optionally substituted C1-C4 alkyl, C2-C4 optionally substituted alkenyl, optionally substituted C2-C4 alkynyl, C(O)R', C(O)OR', C(O)N(R')2, S(O)R', S(O)2R', or S(O)2N(R')2; each R' is independently H or optionally substituted C1-C4 alkyl; Y1 is C, CH, or N; Y2, Y3, Y4, and Y7 are independently C or N; Y5 is CH, CH2, or N; Y6 is C(O), CH, CH2, or N; R2 is absent, is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl; R3 is absent, or R2 and R3 combine with the atom to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or optionally substituted 3- to 14-membered heterocycloalkyl; R4 is absent, is hydrogen, halogen, cyano, or methyl optionally substituted by 1 to 3 halogens; R5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl, or cyclobutyl; R6 is hydrogen or methyl; R7 is hydrogen, halogen, or optionally substituted C1-C3 alkyl, or R6 and R7 combine with the carbon atoms to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7 and R8 combine with the carbon atom to which they are attached to form C=CR7'R8'; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;R7a and R8a are independently hydrogen, halo, C1-C3 alkyl, or combine with the carbon to which they are attached to form a carbonyl;R7' is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R8' is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7' and R8' combine with the carbon atom to which they are attached to form optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R9 is H, F optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; or R9 and L combine with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl; R9' is hydrogen or optionally substituted C1-C6 alkyl; or R9 and R9', combined with the atoms to which they are attached, form a 3- to 6-membered cycloalkyl or a 3- to 6-membered heterocycloalkyl; R10 is hydrogen, halo, hydroxy, C1-C3 alkoxy, or C1-C3 alkyl; R10a is hydrogen or halo; R11 is hydrogen or C1-C3 alkyl; R21 is hydrogen or C1-C3 alkyl (e.g., methyl); and Xe and Xf are, independently, N or CH.

[00104] In some embodiments, a compound of the present invention has the structure of Formula VIa, or a pharmaceutically acceptable salt thereof: Formula VIawherein A is optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene or (e.g., phenyl or phenol) or optionally substituted 5- to 6-membered heteroarylene;B is -CH(R9)- where the carbon is bonded to the carbonyl carbon of NHC(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene or 5- to 6-membered heteroarylene;L is absent or is a ligand;W is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone or an alkynyl sulfone;X1 is optionally substituted C1-C2 alkylene, NR, O, or S(O)n;X2 is O or NH; n is 0, 1, or 2; R is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, C(O)R', C(O)OR', C(O)N(R')2, S(O)R', S(O)2R', or S(O)2N(R')2; each R' is independently H or optionally substituted C1-C4 alkyl; R2 is C1-C6 alkyl, C1-C6 fluoroalkyl, or 3- to 6-membered cycloalkyl; R7 is C1-C3 alkyl; R8 is C1-C3 alkyl; eR9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;Xe and Xf are independently N or CH;R11 is hydrogen or C1-C3 alkyl; andR21 is hydrogen or C1-C3 alkyl.

[00105] In some embodiments of a compound of the present invention, Xe is N and Xf is CH. In some embodiments, Xe is CH and Xf is N.

[00106] In some embodiments, a compound of the present invention has the structure of Formula VIb, or a pharmaceutically acceptable salt thereof: Formula VIb wherein A is optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene or (e.g., phenyl or phenol) or optionally substituted 5- to 6-membered heteroarylene; B is -CH(R9)- where the carbon is bonded to the carbonyl carbon of NHC(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene or 5- to 6-membered heteroarylene; R9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; L is absent or is a ligand; and W is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone, or an alkynyl sulfone. In some embodiments of a compound of the present invention, A is optionally substituted 6-membered arylene.

[00107] In some embodiments, a compound of the present invention has the structure of Formula VIc (corresponding to Formula BB of FIG. 1A and FIG. 1B), or a pharmaceutically acceptable salt thereof: Formula VIcwhere the dotted lines represent zero, one, two, three, or four nonadjacent double bonds;A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene (e.g., phenyl or phenol), or optionally substituted 5- to 10-membered heteroarylene;B is absent, is -CH(R9)-, >C=CR9R9', or >CR9R9' where the carbon is bonded to the carbonyl carbon of -N(R11)C(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;G is Optionally substituted C1-C4 alkylene, optionally substituted C1-C4 alkenylene, optionally substituted C1-C4 heteroalkylene, -C(O)O-CH(R6)- where C is attached to -C(R7R8)-, -C(O)NH-CH(R6)- where C is attached to -C(R7R8)-, optionally substituted C1-C4 heteroalkylene, or 3- to 8-membered heteroarylene;L is absent or is a ligand;W is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone, a haloacetal, or an alkynyl sulfone;X1 is optionally substituted C1-C2 alkylene, NR, O, or S(O)n;X2 is O or NH;X3 is N or CH;n is 0, 1, or 2;R is hydrogen, cyano, optionally substituted C1-C4 alkyl, C2-C4 optionally substituted alkenyl, optionally substituted C2-C4 alkynyl, C(O)R', C(O)OR', C(O)N(R')2, S(O)R', S(O)2R', or S(O)2N(R')2; each R' is independently H or optionally substituted C1-C4 alkyl;Y 1 is C, CH, or N;Y 2, Y3, Y4, and Y7 are independently C or N;Y 5 is CH, CH2, or N;Y6 is C(O), CH, CH2, or N;R2 is absent, is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl; R3 is absent, or R2 and R3 combine with the atom to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or optionally substituted 3- to 14-membered heterocycloalkyl; R4 is absent, is hydrogen, halogen, cyano, or methyl optionally substituted by 1 to 3 halogens; R5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl, or cyclobutyl; R6 is hydrogen or methyl; R7 is hydrogen, halogen, or optionally substituted C1-C3 alkyl, or R6 and R7 combine with the carbon atoms to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7 and R8 combine with the carbon atom to which they are attached to form C=CR7'R8'; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;R7a and R8a are independently hydrogen, halo, C1-C3 alkyl, or combine with the carbon to which they are attached to form a carbonyl;R7' is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R8' is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7' and R8' combine with the carbon atom to which they are attached to form optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R9 is H, F optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; or R9 and L combine with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl; R9' is hydrogen or optionally substituted C1-C6 alkyl; or R9 and R9', combined with the atoms to which they are attached, form a 3- to 6-membered cycloalkyl or a 3- to 6-membered heterocycloalkyl; R10 is hydrogen, halo, hydroxy, C1-C3 alkoxy, or C1-C3 alkyl; R10a is hydrogen or halo; R11 is hydrogen or C1-C3 alkyl; and R21 is hydrogen or C1-C3 alkyl (e.g., methyl). In some embodiments, A has the structure: wherein R13 is hydrogen, halo, hydroxy, amino, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; and R13a is hydrogen or halo. In some embodiments, R13 is hydrogen. In some embodiments, R13 and R13a are each hydrogen. In some embodiments, R13 is hydroxy, methyl, fluoro, or difluoromethyl.

[00108] In some embodiments, A is optionally substituted 5- to 6-membered heteroarylene. In some embodiments, A is:

[00109] In some embodiments, A is optionally substituted C1-C4 heteroalkylene. In some embodiments, A is: . In some embodiments, A is optionally substituted 3- to 6-membered heterocycloalkylene. In some embodiments, A is: . In some modalities, A is

[00110] In some embodiments of a compound of the present invention, B is -CHR9-. In some embodiments, R9 is H, F, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl. In some embodiments, R9 is: . In some modalities, R9 is: In some embodiments, R9 is H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl.

[00111] In some embodiments of a compound of the present invention, B is optionally substituted 6-membered arylene. In some embodiments, B is 6-membered arylene. In some embodiments, B is:

[00112] In some embodiments of a compound of the present invention, R7 is methyl.

[00113] In some embodiments of a compound of the present invention, R8 is methyl.

[00114] In some embodiments, R21 is hydrogen.

[00115] In some embodiments of a compound of the present invention, the linker is the structure of Formula II: A1-(B1)f-(C1)g-(B2)h-(D1)-(B3)i-(C2)j-(B4)k-A2Formula IIwhere A1 is a bond between the linker and B; A2 is a bond between W and the linker; B1, B2, B3, and B4 each, independently, are selected from optionally substituted C1-C2 alkylene, optionally substituted C1-C3 heteroalkylene, O, S, and NRN; RN is hydrogen, optionally substituted C1-4 alkyl, C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted C1-C7 heteroalkyl; C1 and C2 are each independently selected from carbonyl, thiocarbonyl, sulfonyl, or phosphoryl; f, g, h, i, j, and k are each independently 0 or 1; and D1 is optionally substituted C1-C10 alkylene, optionally substituted C2-C10 alkenylene, optionally substituted C2-C10 alkynylene, optionally substituted 3- to 14-membered heterocycloalkylene, optionally substituted 5- to 10-membered heteroarylene, optionally substituted 3- to 8-membered cycloalkylene, optionally substituted 6- to 10-membered arylene, optionally substituted C2-C10 polyethylene glycolene, or optionally substituted C1-C10 heteroalkylene, or a chemical bond A1-(B1)f-(C1)g-(B2)h- a -(B3)i-(C2)j-(B4)k-A2 In some embodiments, the linker is acyclic. In some embodiments, the linker has the structure of Formula IIa: Formula IIa wherein Xa is absent or is N; R14 is absent, is hydrogen or optionally substituted C1-C6 alkyl; and L2 is absent, -SO2-, optionally substituted C1-C4 alkylene or optionally substituted C1-C4 heteroalkylene, wherein at least one of Xa, R14, or L2 is present. In some embodiments, the ligand has the structure:

[00116] In some embodiments, the linker is or comprises a cyclic moiety. In some embodiments, the linker has the structure of Formula IIb: Formula IIwhere o is 0 or 1;R15 is hydrogen, or optionally substituted C1-C6 alkyl, optionally substituted 3- to 8-membered cycloalkylene, or optionally substituted 3- to 8-membered heterocycloalkylene;X4 is absent, is optionally substituted C1-C4 alkylene, O, NCH3, or optionally substituted C1-C4 heteroalkylene;Cy is optionally substituted 3- to 8-membered cycloalkylene, optionally substituted 3- to 8-membered heterocycloalkylene, optionally substituted 6- to 10-membered arylene, or optionally substituted 5- to 10-membered heteroarylene;L3 is absent, is -SO2-, optionally substituted C1-C4 alkylene, or optionally substituted C1-C4 heteroalkylene.

[00117] . In some embodiments, the ligand has the structured Formula IIb-1: Formula IIb-1where o is 0 or 1;R15 is hydrogen, or optionally substituted C1-C6 alkyl, optionally substituted 3- to 8-membered cycloalkylene, or optionally substituted 3- to 8-membered heterocycloalkylene;Cy is optionally substituted 3- to 8-membered cycloalkylene, optionally substituted 3- to 8-membered heterocycloalkylene, optionally substituted 6- to 10-membered arylene, or optionally substituted 5- to 10-membered heteroarylene;L3 is absent, is -SO2-, optionally substituted C1-C4 alkylene, or optionally substituted C1-C4 heteroalkylene.

[00118] In some embodiments, the ligand has the structured Formula IIc: wherein R15 is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted 3- to 8-membered cycloalkylene, or optionally substituted 3- to 8-membered heterocycloalkylene; and R15a, R15b, R15c, R15d, R15e, R15f, and R15g are independently hydrogen, halo, hydroxy, cyano, amino, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, or R15b and R15d combine with the carbons to which they are attached to form an optionally substituted 3- to 8-membered cycloalkylene or optionally substituted 3- to 8-membered heterocycloalkylene.

[00119] In some embodiments, the ligand has the structure:

[00120] In some embodiments, the ligand has the structure:

[00121] In some embodiments, the ligand has the structure

[00122] In some embodiments, the ligand has the structure .In some embodiments of a compound of the present invention, W is a crosslinking group comprising a vinyl ketone. In some embodiments, W has the structure of Formula IIIa: Formula IIIa wherein R16a, R16b, and R16c are independently hydrogen, -CN, halogen, or -C1-C3 alkyl optionally substituted by one or more substituents independently selected from -OH, -O-C1-C3 alkyl, -NH2, -NH(C1-C3 alkyl), -N(C1-C3 alkyl)2, or a 4- to 7-membered saturated heterocycloalkyl. In some embodiments, W is: or In some embodiments, W is a crosslinking group comprising an inone. In some embodiments, W has the structure of Formula IIIb: Formula IIIwhere R17 is hydrogen, -C1-C3 alkyl optionally substituted by one or more substituents independently selected from -OH, -O-C1-C3 alkyl, -NH2, -NH(C1-C3 alkyl), -N(C1-C3 alkyl)2, or a 4- to 7-membered saturated heterocycloalkyl, or a 4- to 7-membered saturated heterocycloalkyl. In some embodiments, W is: . In some modalities, W is

[00123] In some embodiments, W is a crosslinking group comprising a vinyl sulfone. In some embodiments, W has the structure of Formula IIIc: Formula III wherein R18a, R18b, and R18c are independently hydrogen, -CN or -C1-C3 alkyl optionally substituted by one or more substituents independently selected from -OH, -O-C1-C3 alkyl, -NH2, -NH(C1-C3 alkyl), -N(C1-C3 alkyl)2, or a 4- to 7-membered saturated heterocycloalkyl. In some embodiments, W is: or . In some embodiments, W is a crosslinking group comprising an alkynyl sulfone. In some embodiments, W has the structure of Formula IIId: wherein R19 is hydrogen, -C1-C3 alkyl optionally substituted by one or more substituents independently selected from -OH, -O-C1-C3 alkyl, -NH2, -NH(C1-C3 alkyl), -N(C1-C3 alkyl)2, or a 4- to 7-membered saturated heterocycloalkyl, or a 4- to 7-membered saturated heterocycloalkyl. In some embodiments, W is: . In some embodiments, W has the structure of Formula IIIe: Formula IIIe wherein Xe is a halogen; and R20 is hydrogen, -C1-C3 alkyl optionally substituted with one or more substituents independently selected from -OH, -O-C1-C3 alkyl, -NH2, -NH(C1-C3 alkyl), -N(C1-C3 alkyl)2, or a 4- to 7-membered saturated heterocycloalkyl. In some embodiments, W is haloacetal, In some embodiments, W is not haloacetal.

[00124] In some embodiments, a compound of the present invention is selected from Table 1, or a pharmaceutically acceptable salt or stereoisomer thereof. In some embodiments, a compound of the present invention is selected from Table 1, or a pharmaceutically acceptable salt or atropisomer thereof. TABLE 1: CERTAIN COMPOUNDS OF THE PRESENT INVENTION Note that some compounds are shown with bonds as planar or wedge bonds. In some cases, the relative stereochemistry of the stereoisomers has been determined; in some cases, the absolute stereochemistry has been determined. In some cases, a single Example number corresponds to a mixture of stereoisomers. All stereoisomers of the compounds in the preceding table are contemplated by the present invention. In particular embodiments, an atropisomer of a compound in the preceding table is contemplated. The parentheses are to be ignored.*The activity of this stereoisomer may, in fact, be attributable to the presence of a small amount of the stereoisomer with the (S) configuration at the -NC(O)-CH(CH3)2-N(CH3)- position.

[00125] In some embodiments, a compound of Table 2, or a pharmaceutically acceptable salt thereof, is provided. In some embodiments, a compound of the present invention is selected from Table 2, or a pharmaceutically acceptable salt or atropisomer thereof. TABLE 2: CERTAIN COMPOUNDS OF THE PRESENT INVENTION Note that some compounds are shown with bonds as planar or wedge bonds. In some cases, the relative stereochemistry of the stereoisomers has been determined; in some cases, the absolute stereochemistry has been determined. All stereoisomers of the compounds in the preceding table are contemplated by the present invention. In particular embodiments, an atropisomer of a compound in the preceding table is contemplated.

[00126] In some embodiments, a compound of the present invention is or acts as a prodrug, such as with respect to administration to a cell or to a subject in need thereof.

[00127] Also provided are pharmaceutical compositions comprising a compound of the present invention, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[00128] Further provided is a conjugate, or salt thereof, comprising the structure of Formula IV:ML-PFormula IV wherein L is a ligand; P is a monovalent organic moiety; and M has the structure of Formula Va: Formula Where the dotted lines represent zero, one, two, three, or four nonadjacent double bonds; A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 6-membered heteroarylene; B is absent, is -CH(R9)-, >C=CR9R9', or >CR9R9' where the carbon is bonded to the carbonyl carbon of N(R11)C(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene; G is optionally substituted C1-C4 alkylene, optionally substituted C1-C4 alkenylene, optionally substituted C1-C4 heteroalkylene, -C(O)O-CH(R6)- where C is attached to -C(R7R8)-, -C(O)NH-CH(R6)- where C is attached to -C(R7R8)-, optionally substituted C1-C4 heteroalkylene, or 3- to 8-membered heteroarylene; X 1 is optionally substituted C 1-C 2 alkylene, NR, O, or S(O) n; X 2 is O or NH; X 3 is N or CH; n is 0, 1, or 2; R is hydrogen, cyano, optionally substituted C 1-C 4 alkyl, optionally substituted C 2-C 4 alkenyl, optionally substituted C 2-C 4 alkynyl, C(O) R', C(O) OR', C(O) N(R') 2 , S(O) R', S(O) 2R', or S(O) 2N(R') 2 ; each R' is independently H or optionally substituted C 1-C 4 alkyl; Y 1 is C, CH, or N; Y 2 , Y 3 , Y 4, and Y 7 are independently C or N; Y 5 is CH, CH 2 , or N; Y 6 is C(O), CH, CH 2 , or N; R 1 is cyano, optionally substituted C 1-C 6 alkyl, Optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl, or R1 and R2 combine with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl; R2 is absent, is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl; R3 is absent, or R2 and R3 combine with the atom to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or optionally substituted 3- to 14-membered heterocycloalkyl; R4 is absent, is hydrogen, halogen, cyano, or methyl optionally substituted by 1 to 3 halogens; R5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl, or cyclobutyl; R6 is hydrogen or methyl; R7 is hydrogen, halogen, or optionally substituted C1-C3 alkyl, or R6 and R7 combine with the carbon atoms to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7 and R8 combine with the carbon atom to which they are attached to form C=CR7'R8'; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;R7a and R8a are independently hydrogen, halo, C1-C3 alkyl, or combine with the carbon to which they are attached to form a carbonyl;R7' is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R8' is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7' and R8' combine with the carbon atom to which they are attached to form optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R9 is H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl, or R9 and L combine with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl; R9' is hydrogen or optionally substituted C1-C6 alkyl; or R9 and R9', combined with the atoms to which they are attached, form a 3- to 6-membered cycloalkyl or a 3- to 6-membered heterocycloalkyl; R10 is hydrogen, halo, hydroxy, C1-C3 alkoxy, or C1-C3 alkyl; R10a is hydrogen or halo; and R11 is hydrogen or C1-C3 alkyl. In some embodiments, the conjugate, or salt thereof, comprises the structure of Formula IV: ML-PFormula IVwherein L is a ligand; P is a monovalent organic moiety; and M has the structure of Formula Vb: Formula Vwhere the dotted lines represent zero, one, two, three, or four nonadjacent double bonds;A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 6-membered heteroarylene;B is -CH(R9)- or >C=CR9R9' where the carbon is bonded to the carbonyl carbon of -N(R11)C(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;G is C1-C4 alkylene, optionally substituted C1-C4 alkenylene substituted, optionally substituted C1-C4 heteroalkylene, -C(O)O-CH(R6)- where C is attached to -C(R7R8)-, -C(O)NH-CH(R6)- where C is attached to -C(R7R8)-, optionally substituted C1-C4 heteroalkylene, or 3- to 8-membered heteroarylene;X1 is optionally substituted C1-C2 alkylene, NR, O, or S(O)n;X2 is O or NH;X3 is N or CH; n is 0, 1, or 2; R is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, C(O)R', C(O)OR', C(O)N(R')2, S(O)R', S(O)2R', or S(O)2N(R')2; each R' is independently H or optionally substituted C1-C4 alkyl; Y 1 is C, CH, or N; Y 2, Y3, Y4, and Y7 are independently C or N; Y 5 is CH, CH2, or N; Y6 is C(O), CH, CH2, or N; R 1 is cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl or optionally substituted 5- to 10-membered heteroaryl, or R1 and R2 combine with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl; R2 is absent, is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl; R3 is absent, or R2 and R3 combine with the atom to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or optionally substituted 3- to 14-membered heterocycloalkyl; R4 is absent, is hydrogen, halogen, cyano, or methyl optionally substituted by 1 to 3 halogens; R5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl, or cyclobutyl; R6 is hydrogen or methyl; R7 is hydrogen, halogen, or optionally substituted C1-C3 alkyl, or R6 and R7 combine with the carbon atoms to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7 and R8 combine with the carbon atom to which they are attached to form C=CR7'R8'; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;R7a and R8a are independently hydrogen, halo, C1-C3 alkyl, or combine with the carbon to which they are attached to form a carbonyl;R7' is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R8' is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7' and R8' combine with the carbon atom to which they are attached to form optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl, or R9 and L combine with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl; R9' is hydrogen or optionally substituted C1-C6 alkyl; R10 is hydrogen, halo, hydroxy, C1-C3 alkoxy, or C1-C3 alkyl; R10a is hydrogen or halo; and R11 is hydrogen or C1-C3 alkyl. In some embodiments, the conjugate has the structure of Formula IV: ML-PFormula IVwhere L is a ligand; P is a monovalent organic moiety; and M has the structure of Formula Vc: Formula V with the dotted lines representing zero, one, two, three, or four nonadjacent double bonds;A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 6-membered heteroarylene;B is -CH(R9)- where the carbon is bonded to the carbonyl carbon of -N(R11)C(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;G is optionally substituted C1-C4 alkylene, optionally substituted C1-C4 alkenylene, C1-C4 optionally substituted heteroalkylene, - C(O)O-CH(R6)- where C is attached to -C(R7R8)-, -C(O)NH-CH(R6)- where C is attached to -C(R7R8)-, optionally substituted C1-C4 heteroalkylene, or 3- to 8-membered heteroarylene;X1 is optionally substituted C1-C2 alkylene, NR, O, or S(O)n;X2 is O or NH;X3 is N or CH;n is 0, 1, or 2;R is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, C(O)R', C(O)OR', C(O)N(R')2, S(O)R', S(O)2R', or S(O)2N(R')2;each R' is independently H or Optionally substituted C1-C4 alkyl;Y 1 is C, CH, or N;Y 2, Y3, Y4, and Y7 are independently C or N;Y 5 and Y6 are independently CH or N;R 1 is cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl;R 2 is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl; R3 is absent, or R2 and R3 combine with the atom to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or optionally substituted 3- to 14-membered heterocycloalkyl; R4 is absent, is hydrogen, halogen, cyano, or methyl optionally substituted by 1 to 3 halogens; R5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl, or cyclobutyl; R6 is hydrogen or methyl; R7 is hydrogen, halogen, or optionally substituted C1-C3 alkyl, or R6 and R7 combine with the carbon atoms to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7 and R8 combine with the carbon atom to which they are attached to form C=CR7'R8'; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; R7' is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R8' is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7' and R8' combine with the carbon atom to which they are attached to form optionally substituted 3- to 6-membered cycloalkyl or 3-membered heterocycloalkyl. optionally substituted 7-membered; R9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; R10 is hydrogen, hydroxy, C1-C3 alkoxy, or C1-C3 alkyl; and R11 is hydrogen or C1-C3 alkyl. In some embodiments, a compound of the present invention has the structure of Formula IV: ML-PFormula IVwhere L is a ligand; P is a monovalent organic moiety; and M has the structure of Formula Vd: Formula Vd wherein A is optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene or (e.g., phenyl or phenol) or optionally substituted 5- to 6-membered heteroarylene;B is -CH(R9)- where the carbon is bonded to the carbonyl carbon of NHC(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;X1 is optionally substituted C1-C2 alkylene, NR, O, or S(O)n;X2 is O or NH;n is 0, 1, or 2;R is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, C(O)R', C(O)OR', C(O)N(R')2, S(O)R', S(O)2R', or S(O)2N(R')2; each R' is independently H or optionally substituted C1-C4 alkyl; R2 is C1-C6 alkyl, C1-C6 fluoroalkyl, or 3- to 6-membered cycloalkyl; R7 is C1-C3 alkyl; R8 is C1-C3 alkyl; and R9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; Xe and Xf are independently N or CH; R11 is hydrogen or C1-C3 alkyl; and R21 is hydrogen or C1-C3 alkyl. In some embodiments of a compound of the present invention, Xe is N and Xf is CH. In some embodiments, Xe is CH and Xf is N. In some embodiments, a compound of the present invention has the structure of Formula IV: ML-PFormula IVwhere L is a ligand; P is a monovalent organic moiety; and M has the structure of Formula Ve: Formula We see that A is optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene or (e.g., phenyl or phenol) or optionally substituted 5- to 6-membered heteroarylene; B is -CH(R9)- where the carbon is bonded to the carbonyl carbon of NHC(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene or 5- to 6-membered heteroarylene; and R9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl.

[00129] In some embodiments of a conjugate of the present invention, the linker has the structure of Formula II: A1-(B1)f-(C1)g-(B2)h-(D1)-(B3)i-(C2)j-(B4)k-A2Formula IIwhere A1 is a bond between the linker and B; A2 is a bond between P and the linker; B1, B2, B3, and B4 are each independently selected from optionally substituted C1-C2 alkylene, optionally substituted C1-C3 heteroalkylene, O, S, and NRN; RN is hydrogen, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted C1-C7 heteroalkyl; C1 and C2 are each independently selected from carbonyl, thiocarbonyl, sulfonyl, or phosphoryl; f, g, h, i, j, and k are each independently 0 or 1; and D1 is optionally substituted C1-C10 alkylene, optionally substituted C2-C10 alkenylene, optionally substituted C2-C10 alkynylene, optionally substituted 3- to 14-membered heterocycloalkylene, optionally substituted 5- to 10-membered heteroarylene, optionally substituted 3- to 8-membered cycloalkylene, optionally substituted 6- to 10-membered arylene, optionally substituted C2-C10 polyethylene glycolene, or optionally substituted C1-C10 heteroalkylene, or a chemical bond A1-(B1)f-(C1)g-(B2)h- a -(B3)i-(C2)j-(B4)k-A2.

[00130] In some embodiments of a conjugate of the present invention, the monovalent organic moiety is a protein, such as a Ras protein. In some embodiments, the Ras protein is K-Ras G12C, K-Ras G13C, H-Ras G12C, H-Ras G13C, N-Ras G12C, or N-Ras G13C. Other RAS proteins are described herein. In some embodiments, the linker is attached to the monovalent organic moiety via a bond to a sulfhydryl group of an amino acid residue of the monovalent organic moiety. In some embodiments, the linker is attached to the monovalent organic moiety via a bond to a carboxyl group of an amino acid residue of the monovalent organic moiety.

[00131] Also provided is a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof. The cancer may, for example, be pancreatic cancer, colorectal cancer, non-small cell lung cancer, acute myeloid leukemia, multiple myeloma, adenocarcinoma of the thyroid gland, myelodysplastic syndrome, or squamous cell lung carcinoma. In some embodiments, the cancer comprises a Ras mutation, such as K-Ras G12C, K-Ras G13C, H-Ras G12C, H-Ras G13C, N-Ras G12C, or N-Ras G13C. Other Ras mutations are described herein.

[00132] Further provided is a method of treating a Ras protein-related disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof.

[00133] Further provided is a method of inhibiting a Ras protein in a cell, the method comprising contacting the cell with an effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof. For example, the Ras protein is K-Ras G12C, K-Ras G13C, H-Ras G12C, H-Ras G13C, N-Ras G12C, or N-Ras G13C. Other RAS proteins are described herein. The cell may be a cancer cell, such as a pancreatic cancer cell, a colorectal cancer cell, a non-small cell lung cancer cell, an acute myeloid leukemia cell, a multiple myeloma cell, a thyroid gland adenocarcinoma cell, a myelodysplastic syndrome cell, or a squamous cell lung carcinoma cell. Other types of cancer are described herein. The cell may be in vivo or in vitro.

[00134] With respect to the compounds of the present invention, one stereoisomer may exhibit better inhibition than another stereoisomer. For example, one atropisomer may exhibit inhibition, while the other atropisomer may exhibit little or no inhibition.

[00135] In some embodiments, a method or use described herein further comprises administering an additional anti-cancer therapy. In some embodiments, the additional anti-cancer therapy is a HER2 inhibitor, an EGFR inhibitor, a second Ras inhibitor, a SHP2 inhibitor, a SOS1 inhibitor, a Raf inhibitor, a MEK inhibitor, an ERK inhibitor, a PI3K inhibitor, a PTEN inhibitor, an AKT inhibitor, an mTORC1 inhibitor, a BRAF inhibitor, a PD-L1 inhibitor, a PD-1 inhibitor, a CDK4/6 inhibitor, or a combination thereof. In some embodiments, the additional anti-cancer therapy is a SHP2 inhibitor. Other additional anti-cancer therapies are described herein.

SYNTHESIS METHODS

[00136] The compounds described herein can be made from commercially available raw materials or synthesized using known organic, inorganic, or enzymatic processes.

[00137] The compounds of the present invention may be prepared in a variety of ways known to one skilled in the art of organic synthesis. By way of example, the compounds of the present invention may be synthesized using the methods described in the Schemes below together with synthetic methods known in the art of organic chemistry or variations thereof as appreciated by those skilled in the art. Such preferred methods include, but are not limited to, the methods described in the Schemes below. SCHEME 1. GENERAL SYNTHESIS OF MACROCYCLIC ESTERS

[00138] A general synthesis of macrocyclic esters is outlined in Scheme 1. An appropriately substituted aryl-3-(5-bromo-1-ethyl-1H-indol-3-yl)-2,2-dimethylpropan-1-ol (1) can be prepared in three steps from protected 3-(5-bromo-2-iodo-1H-indol-3-yl)-2,2-dimethylpropan-1-ol and appropriately substituted boronic acid, including palladium-mediated coupling, alkylation, and deprotection reactions. Methyl-amino-hexahydropyridazine-3-carboxylate-boronic ester (2) can be prepared in three steps, including protection, iridium catalyst-mediated biorylation, and coupling with methyl (S)-hexahydropyridazine-3-carboxylate.

[00139] An appropriately substituted acetylpyrrolidine-3-carbonyl-N-methyl-L-valine (or an alternative amino acid derivative (4)) can be made by coupling methyl-L-valinate and protected (S)-pyrrolidine-3-carboxylic acid, followed by deprotection, coupling with a carboxylic acid containing an appropriately substituted Michael acceptor, and a hydrolysis step.

[00140] The final macrocyclic esters can be made by coupling methyl-amino-hexahydropyridazine-3-carboxylate-boronic ester (2) and aryl-3-(5-bromo-1-ethyl-1H-indol-3-yl)-2,2-dimethylpropan-1-ol (1) in the presence of a Pd catalyst followed by hydrolysis and macrolactonization steps to afford a suitably protected macrocyclic intermediate (5). Deprotection and coupling with an appropriately substituted intermediate 4 results in a macrocyclic product. Additional deprotection and/or functionalization steps may be required to afford the final compound. SCHEME 2. ALTERNATIVE GENERAL SYNTHESIS OF MACROCYCLIC ESTERS

[00141] Alternatively, the macrocyclic ester can be prepared as described in Scheme 2. A suitably protected bromoindolyl (6) coupled in the presence of a Pd catalyst with boronic ester (3), followed by iodination, deprotection and ester hydrolysis. Subsequent coupling with methyl (S)-hexahydropyridazine-3-carboxylate, followed by hydrolysis and macrolactonization can result in iodine intermediate (7). Coupling in the presence of a Pd catalyst with an appropriately substituted boronic ester and alkylation can produce fully protected macrocycle (5). Additional deprotection or functionalization steps are required to produce the final compound.

[00142] Furthermore, compounds of the disclosure may be synthesized using the methods described in the Examples below, together with synthetic methods known in the art of synthetic organic chemistry, or variations thereof, as appreciated by those skilled in the art. Such preferred methods include, but are not limited to, the methods described in the Examples below. For example, one of skill in the art would be able to install into a macrocyclic ester a desired -B-L-W group of a compound of Formula (I), where B, L, and W are defined herein, including using methods exemplified in the Example section herein.

[00143] The compounds in Table 1 herein were prepared using methods disclosed herein or were prepared using methods disclosed herein combined with the knowledge of one skilled in the art. The compounds in Table 2 can be prepared using methods disclosed herein or can be prepared using methods disclosed herein combined with the knowledge of one skilled in the art. SCHEME 3. GENERAL SYNTHESIS OF MACROCYCLIC ESTERS

[00144] An alternative general synthesis of macrocyclic esters is outlined in Scheme 3. An appropriately substituted indolyl boronic ester (8) can be prepared in four steps from protected 3-(5-bromo-2-iodo-1H-indol-3-yl)-2,2-dimethylpropan-1-ol and appropriately substituted boronic acid, including palladium-mediated biorylation, alkylation, deprotection, and palladium-mediated biorylation reactions.

[00145] Methyl-amino-3-(4-bromothiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylate (10) can be prepared by coupling (S)-2-amino-3-(4-bromothiazol-2-yl)propanoic acid (9) with methyl (S)-hexahydropyridazine-3-carboxylate.

[00146] The final macrocyclic esters can be made by coupling methyl-amino-3-(4-bromothiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylate (10) and an appropriately substituted indolyl boronic ester (8) in the presence of Pd catalyst followed by hydrolysis and macrolactonization steps to afford a suitably protected macrocyclic intermediate (11). Deprotection and coupling with an appropriately substituted intermediate 4 can afford a macrocyclic product. Additional deprotection or functionalization steps may be required to afford a final compound 13 or 14. SCHEME 4. GENERAL SYNTHESIS OF MACROCYCLIC ESTERS

[00147] An alternative general synthesis of macrocyclic esters is outlined in Scheme 4. An appropriately substituted morpholine or alternative herecyclic intermediate (15) can be coupled to suitably protected Intermediate 1 via palladium-mediated coupling. Subsequent ester hydrolysis and coupling with piperazoic ester affords intermediate 16.

[00148] Macrocyclic esters can be made by a sequence of hydrolysis, deprotection, and macrocyclization. Subsequent deprotection and coupling with Intermediate 4 (or analogs) results in appropriately substituted macrocyclic final products. Additional deprotection or functionalization steps may be required to produce a final compound 17. SCHEME 5. GENERAL SYNTHESIS OF MACROCYCLIC ESTERS

[00149] An alternative general synthesis of macrocyclic esters is outlined in Scheme 5. An appropriately substituted macrocycle (20) can be prepared from an appropriately protected boronic ester 18 and bromoindolyl intermediate (19), including palladium-mediated coupling, hydrolysis, piperazoic ester coupling, hydrolysis, deprotection, and macrocyclization steps. Subsequent coupling with an appropriately substituted protected amino acid is followed by palladium-mediated coupling intermediate 21. Additional deprotection and derivatization steps, including alkylation, may be required at this point.

[00150] The final macrocyclic esters can be made by coupling intermediate (22) and an appropriately substituted carboxylic acid intermediate (23). Additional deprotection or functionalization steps may be necessary to produce a final compound (24).

[00151] Furthermore, compounds of the disclosure may be synthesized using the methods described in the Examples below, together with synthetic methods known in the art of synthetic organic chemistry, or variations thereof, as appreciated by those skilled in the art. Such preferred methods include, but are not limited to, the methods described in the Examples below. For example, one of skill in the art would be able to install into a macrocyclic ester a desired -B-L-W group of a compound of Formula (I), where B, L, and W are defined herein, including using methods exemplified in the Example section herein.

PHARMACEUTICAL COMPOSITIONS AND METHODS OF USE PHARMACEUTICAL COMPOSITIONS AND METHODS OF ADMINISTRATION

[00152] The compounds with which the invention is concerned are inhibitors of Ras and are useful in the treatment of cancer. Accordingly, one embodiment of the present invention provides pharmaceutical compositions containing a compound of the invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient, as well as methods of using the compounds of the invention to prepare such compositions.

[00153] As used herein, the term “pharmaceutical composition” refers to a compound, such as a compound of the present invention, or a pharmaceutically acceptable salt thereof, formulated together with a pharmaceutically acceptable excipient.

[00154] In some embodiments, a compound is present in a pharmaceutical composition in a unit dose amount appropriate for administration in a therapeutic regimen that shows a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, the pharmaceutical compositions can be specially formulated for administration in solid or liquid form, including those adapted for the following: oral administration, e.g., drenches (aqueous or non-aqueous solutions or suspensions); tablets, e.g., those intended for buccal, sublingual, and systemic absorption, boluses, powders, granules, pastes for application to the tongue; parenteral administration, e.g., by subcutaneous, intramuscular, intravenous, or epidural injection as, e.g., a sterile solution or suspension, or sustained-release formulation; topical application, e.g., as a cream, ointment, or a controlled-release patch or spray applied to the skin, lungs, or oral cavity; intravaginally or intrarectally, for example, as a pessary, cream, or foam; sublingually; ocularly; transdermally; or nasally, pulmonaryly, and to other mucosal surfaces.

[00155] A “pharmaceutically acceptable excipient,” as used herein, refers to any inactive ingredient (e.g., a carrier capable of suspending or dissolving the active compound) with the properties of being non-toxic and non-inflammatory in a subject. Typical excipients may include, for example: anti-adherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, sorbents, suspending or dispersing agents, sweeteners, or waters of hydration. Excipients include, but are not limited to: optionally substituted butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, cross-linked polyvinylpyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, optionally substituted hydroxypropyl cellulose, optionally substituted hydroxypropyl methylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methyl paraben, microcrystalline cellulose, polyethylene glycol, polyvinyl pyrrolidone, povidone, pregelatinized starch, propyl paraben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, stearic acid, sucrose, talc, carbon dioxide titanium, vitamin A, vitamin E, vitamin C, and xylitol. Those skilled in the art are familiar with a variety of agents and materials useful as excipients. See, e.g., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, et al., Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005. In some embodiments, a composition includes at least two different pharmaceutically acceptable excipients.

[00156] The compounds described herein, whether expressly stated or not, may be provided or used in salt form, e.g., a pharmaceutically acceptable salt form, unless expressly stated otherwise. The term “pharmaceutically acceptable salt,” as used herein, refers to those salts of the compounds described herein that are, within the scope of medical judgment, suitable for use in contact with the tissues of humans and other animals without undue toxicity, irritation, allergic response, and the like, and are proportionate to a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in: Berge et al., J. Pharmaceutical Sciences 66:1-19, 1977 and in Pharmaceutical Salts: Properties, Selection, and Use, (Eds. P.H. Stahl and C.G. Wermuth), Wiley-VCH, 2008. Salts can be prepared in situ during the final isolation and purification of the compounds described herein or separately by reaction of the free base group with a suitable organic acid.

[00157] The compounds of the invention may have ionizable groups so as to be capable of preparation as pharmaceutically acceptable salts. Such salts may be acid addition salts involving inorganic or organic acids or the salts may, in the case of acidic forms of the compounds of the invention, be prepared from inorganic or organic bases. In some embodiments, the compounds are prepared or used as pharmaceutically acceptable salts prepared as addition products of pharmaceutically acceptable acids or bases. Suitable pharmaceutically acceptable acids and bases are well known in the art, such as hydrochloric, sulfuric, hydrobromic, acetic, lactic, citric or tartaric acids to form acid addition salts and potassium hydroxide, sodium hydroxide, ammonium hydroxide, caffeine, various amines and the like to form base salts. Methods for preparing the appropriate salts are well established in the art.

[00158] Representative acid addition salts include acetate, adipate salts, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptanate, hexanoate, hydrobromide, hydrochloride, hydroiodide, optionally substituted 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate salts, and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like, as well as non-toxic ammonium, quaternary ammonium, and amine cations, including but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like.

[00159] As used herein, the term a “subject” refers to any member of the animal kingdom. In some embodiments, “subject” refers to humans, at any stage of development. In some embodiments, “subject” refers to a human patient. In some embodiments, “subject” refers to non-human animals. In some embodiments, the non-human animal is a mammal (e.g., a rodent, a mouse, a rat, a rabbit, a monkey, a dog, a cat, a sheep, cattle, a primate, or a pig). In some embodiments, subjects include, but are not limited to, mammals, birds, reptiles, amphibians, fish, or worms. In some embodiments, a subject can be a transgenic animal, genetically modified animal, or a clone.

[00160] As used herein, the term “dosage form” refers to a physically distinct unit of a compound (e.g., a compound of the present invention) for administration to a subject. Each unit contains a predetermined amount of compound. In some embodiments, such amount is a unitary dosage amount (or an integral fraction thereof) suitable for administration according to a dosage regimen that has been determined to correlate with a desired or beneficial outcome when administered to a relevant population (i.e., with a therapeutic dosage regimen). Those skilled in the art appreciate that the total amount of a therapeutic composition or compound administered to a particular subject is determined by one or more attending physicians, and may involve administering multiple dosage forms.

[00161] As used herein, the term “dosing regimen” refers to a set of unit doses (typically more than one) that are administered individually to a subject, typically separated by time periods. In some embodiments, a given therapeutic compound (e.g., a compound of the present invention) has a recommended dosing regimen, which may involve one or more doses. In some embodiments, a dosing regimen comprises a plurality of doses, all of which are separated from each other by a time period of the same duration; in some embodiments, a dosing regimen comprises a plurality of doses and at least two different time periods separating individual doses. In some embodiments, all doses within a dosing regimen are of the same unit dose amount. In some embodiments, different doses within a dosing regimen are of different amounts. In some embodiments, a dosing regimen comprises a first dose at a first dose amount, followed by one or more additional doses at a second dose amount different from the first dose amount. In some embodiments, a dosage regimen comprises a first dose at a first dose amount, followed by one or more additional doses at a second dose amount of the same dose amount as the first dose amount. In some embodiments, a dosage regimen is correlated with a desired or beneficial outcome when administered across a relevant population (i.e., is a therapeutic dosage regimen).

[00162] A “therapeutic regimen” refers to a dosing regimen whose administration across a relevant population is correlated with a desired or beneficial therapeutic outcome.

[00163] The term “treatment” (also “treat” or “treating”), in its broadest sense, refers to any administration of a substance (e.g., a compound of the present invention) that partially or completely alleviates, ameliorates, alleviates, inhibits, delays the onset of, reduces the severity of, or reduces the incidence of one or more symptoms, features, or causes of a specific disease, disorder, or condition. In some embodiments, such treatment may be administered to a subject who exhibits no signs of the relevant disease, disorder, or condition or to a subject who exhibits only early signs of the disease, disorder, or condition. Alternatively, or additionally, in some embodiments, the treatment may be administered to a subject who exhibits one or more established signs of the relevant disease, disorder, or condition. In some embodiments, the treatment may be of a subject who has been diagnosed as suffering from the relevant disease, disorder, or condition. In some embodiments, the treatment may be of a subject known to have one or more susceptibility factors that are statistically correlated with increased risk of developing the relevant disease, disorder, or condition.

[00164] The term “therapeutically effective amount” means an amount that is sufficient, when administered to a population suffering from or susceptible to a disease, disorder, or condition in accordance with a therapeutic dosage regimen, to treat the disease, disorder, or condition. In some embodiments, a therapeutically effective amount is one that reduces the incidence or severity or delays the onset of one or more symptoms of the disease, disorder, or condition. Those skilled in the art will appreciate that the phrase “therapeutically effective amount” does not, in fact, require that successful treatment be achieved in a particular individual. Rather, a therapeutically effective amount may be the amount that provides a particular desired pharmacological response in a significant number of subjects when administered to patients in need of such treatment. It is specifically understood that certain subjects may, in fact, be “refractory” to a “therapeutically effective amount.” In some embodiments, reference to a therapeutically effective amount may be a reference to an amount measured in one or more specific tissues (e.g., a tissue affected by the disease, disorder, or condition) or fluids (e.g., blood, saliva, serum, sweat, tears, urine). Those of skill in the art will appreciate that in some embodiments, a therapeutically effective amount may be formulated or administered in a single dose. In some embodiments, a therapeutically effective amount may be formulated or administered in a plurality of doses, for example, as part of a dosing regimen.

[00165] For use as a treatment of subjects, the compounds of the invention, or a pharmaceutically acceptable salt thereof, may be formulated as pharmaceutical or veterinary compositions. Depending on the subject to be treated, the mode of administration, and the type of treatment desired, e.g., prevention, prophylaxis, or therapy, the compounds, or a pharmaceutically acceptable salt thereof, are formulated in ways consistent with these parameters. A summary of such techniques may be found in Remington: The Science and Practice of Pharmacy, 21st Edition, Lippincott Williams & Wilkins, (2005); and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J. C. Boylan, 1988-1999, Marcel Dekker, New York, each of which is incorporated herein by reference.

[00166] The compositions can be prepared according to conventional mixing, granulating or coating methods, respectively, and the present pharmaceutical compositions can contain from about 0.1% to about 99%, from about 5% to about 90%, or from about 1% to about 20% of a compound of the present invention, or a pharmaceutically acceptable salt thereof, by weight or volume. In some embodiments, the compounds, or a pharmaceutically acceptable salt thereof, described herein can be present in amounts totaling 1-95% by weight of the total weight of a composition, such as a pharmaceutical composition.

[00167] The composition may be provided in a fingertip form that is suitable for intra-articular, oral, parenteral (e.g., intravenous, intramuscular), rectal, cutaneous, subcutaneous, topical, transdermal, sublingual, nasal, vaginal, intravesicular, intraurethral, intrathecal, epidural, aural, or ocular administration, or by injection, inhalation, or direct contact with the nasal, genitourinary, reproductive, or oral mucosa. Thus, the pharmaceutical composition may be in the form of, for example, tablets, capsules, pills, powders, granules, suspensions, emulsions, solutions, gels, including hydrogels, pastes, ointments, creams, patches, salves, osmotic delivery devices, suppositories, enemas, injectables, implants, sprays, preparations suitable for iontophoretic delivery, or aerosols. The compositions may be formulated in accordance with conventional pharmaceutical practice.

[00168] As used herein, the term “administration” refers to the administration of a composition (e.g., a compound, or a preparation comprising a compound as described herein) to a subject or system. Administration to an animal subject (e.g., a human) may be by any appropriate route. For example, in some embodiments, administration may be bronchial (including by bronchial instillation), buccal, enteral, interdermal, intraarterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (including by intratracheal instillation), transdermal, vaginal, or vitreous.

[00169] The formulations may be prepared in a manner suitable for systemic administration or topical or local administration. Systemic formulations include those designed for injection (e.g., intramuscular, intravenous, or subcutaneous injection) or may be prepared for transdermal, transmucosal, or oral administration. A formulation will generally include a diluent, as well as, in some cases, adjuvants, buffers, preservatives, and the like. The compounds, or a pharmaceutically acceptable salt thereof, may also be administered in liposomal compositions or as microemulsions.

[00170] For injection, the formulations may be prepared in conventional forms as liquid solutions or suspensions or as solid forms suitable for solution or suspension in liquid prior to injection or as emulsions. Suitable excipients include, for example, water, saline, dextrose, glycerol and the like. Such compositions may also contain amounts of non-toxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents and the like, such as, for example, sodium acetate, sorbitan monolaurate and the like.

[00171] Various sustained drug release systems have also been devised. See, for example, U.S. Patent 5,624,677.

[00172] Systemic administration may also include relatively non-invasive methods, such as the use of suppositories, transdermal patches, transmucosal delivery, and intranasal administration. Oral administration is also suitable for compounds of the invention, or a pharmaceutically acceptable salt thereof. Suitable forms include syrups, capsules, and tablets, as is understood in the art.

[00173] Each compound, or a pharmaceutically acceptable salt thereof, as described herein may be formulated in a variety of ways that are known in the art. For example, the first and second agents of the combination therapy may be formulated together or separately. Other embodiments of combination therapy are described herein.

[00174] Individually or separately formulated agents may be packaged together as a kit. Non-limiting examples include, but are not limited to, kits containing, for example, two pills, a pill and a powder, a suppository and a liquid in a vial, two topical creams, etc. The kit may include optional components that aid in the administration of the unit dose to subjects, such as vials for reconstitution of powder forms, syringes for injection, custom IV delivery systems, inhalers, etc. In addition, the unit dose kit may contain instructions for the preparation and administration of the compositions. The kit may be manufactured as a single-use unit dose for a subject, multiple uses for a specific subject (at a constant dose or where the individual compounds, or a pharmaceutically acceptable salt thereof, may vary in potency as therapy progresses); or the kit may contain multiple doses suitable for administration to multiple subjects (“bulk packaging”). Kit components can be assembled into boxes, blisters, bottles, tubes and the like.

[00175] Formulations for oral use include tablets containing the active ingredient(s) in admixture with non-toxic pharmaceutically acceptable excipients. These excipients may be, for example, inert diluents or fillers (e.g. sucrose, sorbitol, sugar, mannitol, microcrystalline cellulose, starches including potato starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate or sodium phosphate); granulating and disintegrating agents (e.g. cellulose derivatives including microcrystalline cellulose, starches including potato starch, croscarmellose sodium, alginates or alginic acid); binding agents (e.g., sucrose, glucose, sorbitol, acacia, alginic acid, sodium alginate, gelatin, starch, pregelatinized starch, microcrystalline cellulose, magnesium aluminum silicate, sodium carboxymethyl cellulose, methyl cellulose, optionally substituted hydroxypropyl methyl cellulose, ethyl cellulose, polyvinylpyrrolidone, or polyethylene glycol); and lubricating, glidant, and antiadhesive agents (e.g., magnesium stearate, zinc stearate, stearic acid, silicas, hydrogenated vegetable oils, or talc). Other pharmaceutically acceptable excipients may be colorants, flavoring agents, plasticizers, humectants, buffering agents, and the like.

[00176] Two or more compounds may be mixed in a tablet, capsule, or other vehicle, or may be partitioned. In one example, the first compound is contained inside the tablet and the second compound is on the outside, such that a substantial portion of the second compound is released prior to release of the first compound.

[00177] Formulations for oral use may also be provided as chewable tablets or as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent (e.g. potato starch, lactose, microcrystalline cellulose, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with water or an oil medium, e.g. peanut oil, liquid paraffin or olive oil. Powders, granules and pellets may be prepared using the above-mentioned ingredients into tablets and capsules in a conventional manner using, for example, a mixer, a fluid bed apparatus or spray drying equipment.

[00178] Diffusion-controlled dissolution or release may be achieved by appropriately coating a tablet, capsule, pellet or granulate formulation of compounds, or by incorporating the compound, or a pharmaceutically acceptable salt thereof, into an appropriate matrix. A controlled release coating may include one or more of the above mentioned coating substances or, for example, shellac, beeswax, glycowax, castor wax, carnauba wax, stearyl alcohol, glyceryl monostearate, glyceryl distearate, glycerol palmitostearate, ethyl cellulose, acrylic resins, dl-polylactic acid, cellulose acetate butyrate, polyvinyl chloride, polyvinyl acetate, vinyl pyrrolidone, polyethylene, polymethacrylate, methyl methacrylate, optionally substituted 2-hydroxymethacrylate, methacrylate hydrogels, 1,3-butylene glycol, ethylene glycol methacrylate or polyethylene glycols. In a controlled release matrix formulation, the matrix material may also include, for example, hydrated methyl cellulose, carnauba wax and stearyl alcohol, carbopol 934, silicone, glyceryl tristearate, methyl acrylate-methyl methacrylate, polyvinyl chloride, polyethylene, or halogenated fluorocarbon.

[00179] Liquid forms into which the compounds or a pharmaceutically acceptable salt thereof, and pharmaceutical compositions of the present invention may be incorporated for oral administration include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, and emulsions flavored with edible oils such as cottonseed oil, sesame oil, coconut oil, or peanut oil, as well as elixirs and similar pharmaceutical carriers.

[00180] Generally, when administered to a human, the oral dosage of any of the compounds of the invention, or a pharmaceutically acceptable salt thereof, will depend on the nature of the compound and can be readily determined by one of skill in the art. A dosage can be, for example, about 0.001 mg to about 2000 mg per day, about 1 mg to about 1000 mg per day, about 5 mg to about 500 mg per day, about 100 mg to about 1500 mg per day, about 500 mg to about 1500 mg per day, about 500 mg to about 2000 mg per day, or any range derivable therefrom.

[00181] In some embodiments, the pharmaceutical composition may further comprise an additional compound having antiproliferative activity. Depending on the mode of administration, the compounds, or a pharmaceutically acceptable salt thereof, will be formulated into suitable compositions to allow for easy delivery. Each compound, or a pharmaceutically acceptable salt thereof, of a combination therapy may be formulated in a variety of ways that are known in the art. For example, the first and second agents of the combination therapy may be formulated together or separately. Desirably, the first and second agents are formulated together for simultaneous or nearly simultaneous administration of the agents.

[00182] It will be appreciated that the compounds and pharmaceutical compositions of the present invention may be formulated and employed in combination therapies, that is, the compounds and pharmaceutical compositions may be formulated or administered simultaneously with, prior to, or subsequent to one or more other desired therapeutics or medical procedures. The particular combination of therapies (therapeutics or procedures) to employ in a combination regimen will take into account the compatibility of the desired therapeutics or procedures and the desired therapeutic effect to be achieved. It will also be appreciated that the therapies employed may achieve a desired effect for the same disorder, or may achieve different effects (e.g., control of any adverse effects).

[00183] Administration of each drug in a combination therapy as described herein may independently be from one to four times daily for one day to one year and may even be for the life of the subject. Long-term chronic administration may be indicated.

METHODS OF USE

[00184] In some embodiments, the invention discloses a method of treating a disease or disorder that is characterized by aberrant RAS activity due to a mutant RAS. In some embodiments, the disease or disorder is a cancer.

[00185] Accordingly, there is also provided a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising such a compound or salt. In some embodiments, the cancer is colorectal cancer, non-small cell lung cancer, small cell lung cancer, pancreatic cancer, appendiceal cancer, melanoma, acute myeloid leukemia, small bowel cancer, ampullary cancer, germ cell cancer, cervical cancer, cancer of unknown primary, endometrial cancer, esophagogastric cancer, GI neuroendocrine cancer, ovarian cancer, sex cord stromal tumor cancer, hepatobiliary cancer, or bladder cancer. In some embodiments, the cancer is appendiceal, endometrial, or melanoma. Also provided is a method of treating a Ras protein-related disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising such a compound or salt.

[00186] In some embodiments, the compounds of the present invention or pharmaceutically acceptable salts thereof, pharmaceutical compositions comprising such compounds or salts, and methods provided herein can be used for the treatment of a wide variety of cancers, including tumors such as lung, prostate, breast, brain, skin, cervical carcinomas, testicular carcinomas, etc. More particularly, cancers that can be treated by the compounds or salts thereof, pharmaceutical compositions comprising such compounds or salts, and methods of the invention include, but are not limited to, tumor types such as astrocytic, breast, cervical, colorectal, endometrial, esophageal, gastric, head and neck, hepatocellular, laryngeal, pulmonary, oral, ovarian, prostate, and thyroid carcinomas and sarcomas. Other types of cancer include, for example:Cardiac, for example: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma, and teratoma;Lung, for example: bronchogenic carcinoma (squamous cell, small cell undifferentiated, large cell undifferentiated, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondromatous hamartoma, mesothelioma;Gastrointestinal, for example: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumors, vipoma), small intestine (adenocarcinoma, lymphoma, carcinoid tumors, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large intestine (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma);Genitourinary tract, e.g.: kidney (adenocarcinoma, Wilm's tumor (nephroblastoma), lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma, fibroadenoma, adenomatoid tumors, lipoma);Liver, e.g.: hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma;Biliary tract, e.g., gallbladder carcinoma, ampullary carcinoma, cholangiocarcinoma;Bone, e.g., osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticular cell sarcoma), multiple myeloma, malignant giant cell tumor chordoma, exotic chondrosarcoma, chondroblastoma, chondromyxofibroma, osteoid osteoma, and giant cell tumors;Nervous system, e.g., skull (osteoma, hemangioma, granuloma, xanthoma, osteitis deformans), meninges (meningioma, meningiosarcoma, gliomatosis), brain (astrocytoma, medulloblastoma, glioma, ependymoma, germinoma (pinealoma), glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors), neurofibroma of the spinal cord, neurofibromatosis type 1, meningioma, glioma, sarcoma);Gynecological, for example: uterus (endometrial carcinoma, uterine carcinoma, endometrial carcinoma of the uterine body), cervix (cervical carcinoma, pretumor cervical dysplasia), ovaries (ovarian carcinoma (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa-thecal cell tumors, Sertoli-Leydig cell tumors, dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma), fallopian tubes (carcinoma);Hematological, for example: blood (myeloid leukemia (acute and chronic), acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative diseases, multiple myeloma, myelodysplastic syndrome), Hodgkin's disease, non-Hodgkin's lymphoma (malignant lymphoma);Skin, e.g.: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, soft dysplastic nevi, lipoma, angioma, dermatofibroma, keloids, psoriasis; andAdrenal glands, e.g.: neuroblastoma.

[00187] In some embodiments, the Ras protein is wild-type (RasWT). Accordingly, in some embodiments, a compound of the present invention is employed in a method of treating a patient with a cancer comprising a RasWT (e.g., K-RasWT, H-RasWT, or N-RasWT). In some embodiments, the Ras protein is the Ras amplification (e.g., K-Rasamp). Accordingly, in some embodiments, a compound of the present invention is employed in a method of treating a patient with a cancer comprising a Rasamp (K-Rasamp, H-Rasamp, or N-Rasamp). In some embodiments, the cancer comprises a Ras mutation, such as a Ras mutation described herein. In some embodiments, a mutation is selected from: (a) the following K-Ras mutants: G12D, G12V, G12C, G13D, G12R, G12A, Q61H, G12S, A146T, G13C, Q61L, Q61R, K117N, A146V, G12F, Q61K, L19F, Q22K, V14I, A59T, A146P, G13R, G12L, or G13V, and combinations thereof; (b) the following H-Ras mutants: Q61R, G13R, Q61K, G12S, Q61L, G12D, G13V, G13D, G12C, K117N, A59T, G12V, G13C, Q61H, G13S, A18V, D119N, G13N, A146T, A66T, G12A, A146V, G12N or G12R, and combinations thereof; and (c) the following N-Ras mutants: Q61R, Q61K, G12D, Q61L, Q61H, G13R, G13D, G12S, G12C, G12V, G12A, G13V, G12R, P185S, G13C, A146T, G60E, Q61P, A59D, E132K, E49K, T50I, A146V or A59T, and combinations thereof; or a combination of any of the foregoing. In some embodiments, the cancer comprises a K-Ras mutation selected from the group consisting of G12C, G12D, G13C, G12V, G13D, G12R, G12S, Q61H, Q61K, and Q61L. In some embodiments, the cancer comprises an N-Ras mutation selected from the group consisting of G12C, Q61H, Q61K, Q61L, Q61P, and Q61R. In some embodiments, the cancer comprises an H-Ras mutation selected from the group consisting of Q61H and Q61L. In some embodiments, the cancer comprises a Ras mutation selected from the group consisting of G12C, G13C, G12A, G12D, G13D, G12S, G13S, G12V, and G13V. In some embodiments, the cancer comprises at least two Ras mutations selected from the group consisting of G12C, G13C, G12A, G12D, G13D, G12S, G13S, G12V, and G13V. In some embodiments, a compound of the present invention inhibits more than one Ras mutant. For example, a compound can inhibit K-Ras G12C and K-Ras G13C. A compound can inhibit N-Ras G12C and K-Ras G12C. In some embodiments, a compound can inhibit K-Ras G12C and K-Ras G12D. In some embodiments, a compound can inhibit K-Ras G12V and K-Ras G12C. In some embodiments, a compound can inhibit K-Ras G12V and K-Ras G12S. In some embodiments, a compound of the present invention inhibits RasWT in addition to one or more additional Ras mutations (e.g., K-, H-, or N-RasWT and K-Ras G12D, G12V, G12C, G13D, G12R, G12A, Q61H, G12S, A146T, G13C, Q61L, Q61R, K117N, A146V, G12F, Q61K, L19F, Q22K, V14I, A59T, A146P, G13R, G12L, or G13V; K, H, or N-RasWT and H-Ras Q61R, G13R, Q61K, G12S, Q61L, G12D, G13V, G13D, G12C, K117N, A59T, G12V, G13C, Q61H, G13S, A18V, D119N, G13N, A146T, A66T, G12A, A146V, G12N, or G12R; or K, H or N-RasWT and N-Ras Q61R, Q61K, G12D, Q61L, Q61H, G13R, G13D, G12S, G12C, G12V, G12A, G13V, G12R, P185S, G13C, A146T, G60E, Q61P, A59D, E132K, E49K, T50I, A146V, or A59T). In some embodiments, a compound of the present invention inhibits Rasamp in addition to one or more Ras mutations (e.g., K-, H-, or N-Rasamp and K-Ras G12D, G12V, G12C, G13D, G12R, G12A, Q61H, G12S, A146T, G13C, Q61L, Q61R, K117N, A146V, G12F, Q61K, L19F, Q22K, V14I, A59T, A146P, G13R, G12L, or G13V; K, H, or N-Rasamp and H-Ras Q61R, G13R, Q61K, G12S, Q61L, G12D, G13V, G13D, G12C, K117N, A59T, G12V, G13C, Q61H, G13S, A18V, D119N, G13N, A146T, A66T, G12A, A146V, G12N, or G12R; or K, H or N-Rasamp and N-Ras Q61R, Q61K, G12D, Q61L, Q61H, G13R, G13D, G12S, G12C, G12V, G12A, G13V, G12R, P185S, G13C, A146T, G60E, Q61P, A59D, E132K, E49K, T50I, A146V, or A59T).

[00188] Methods of detecting Ras mutations are known in the art. Such means include, but are not limited to, direct sequencing and use of a high-sensitivity (CE-IVD marked) diagnostic assay, e.g., as described in Domagala, et al., Pol J Pathol 3:145-164 (2012), incorporated herein by reference in its entirety, including TheraScreen PCR; AmoyDx; PNAClamp; RealQuality; EntroGen; LightMix; StripAssay; Hybcell plexA; Devyser; Surveyor; Cobas; and TheraScreen Pyro. See also, e.g., WO 2020/106640.

[00189] In some embodiments, the cancer is non-small cell lung cancer and the Ras mutation comprises a K-Ras mutation, such as K-Ras G12C, K-Ras G12V, or K-Ras G12D. In some embodiments, the cancer is colorectal cancer and the Ras mutation comprises a K-Ras mutation, such as K-Ras G12C, K-Ras G12V, or K-Ras G12D. In some embodiments, the cancer is pancreatic cancer and the Ras mutation comprises a K-Ras mutation, such as K-Ras G12D or K-Ras G12V. In some embodiments, the cancer is pancreatic cancer and the Ras mutation comprises an N-Ras mutation, such as N-Ras G12D. In some embodiments, the cancer is melanoma and the Ras mutation comprises an N-Ras mutation, such as N-Ras Q61R or N-Ras Q61K. In some embodiments, the cancer is non-small cell lung cancer and the Ras protein is K-Rasamp. In any of the foregoing, if not already specified, a compound can inhibit RasWT (e.g., K-, H-, or N-RasWT) or Rasamp (e.g., K, H-, or N-Rasamp) as well.

[00190] In some embodiments, a cancer comprises a Ras mutation and a STK11LOF, a KEAP1, an EPHA5, or an NF1 mutation. In some embodiments, the cancer is non-small cell lung cancer and comprises a K-Ras G12C mutation. In some embodiments, the cancer is non-small cell lung cancer and comprises a K-Ras G12C mutation and a STK11LOF mutation. In some embodiments, the cancer is non-small cell lung cancer and comprises a K-Ras G12C mutation and a STK11LOF mutation. In some embodiments, a cancer comprises a K-Ras G13C Ras mutation and a STK11LOF, a KEAP1, an EPHA5, or an NF1 mutation. In some embodiments, the cancer is non-small cell lung cancer and comprises a K-Ras G12D mutation. In some embodiments, the cancer is non-small cell lung cancer and comprises a K-Ras G12V mutation. In some embodiments, the cancer is colorectal cancer and comprises a K-Ras G12C mutation. In some embodiments, the cancer is pancreatic cancer and comprises a K-Ras G12C or K-Ras G12D mutation. In some embodiments, the cancer is pancreatic cancer and comprises a K-Ras G12V mutation. In some embodiments, the cancer is endometrial cancer, ovarian cancer, cholangiocarcinoma, or appendiceal mucinous cancer and comprises a K-Ras G12C mutation. In some embodiments, the cancer is gastric cancer and comprises a K-Ras G12C mutation. In any of the foregoing, a compound can inhibit RasWT (e.g., K-, H-, or N-RasWT) or Rasamp (e.g., K-, H-, or N-Rasamp) as well.

[00191] Also provided is a method of inhibiting a Ras protein in a cell, the method comprising contacting the cell with an effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof. A method of inhibiting RAF-Ras binding, the method comprising contacting the cell with an effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof, is also provided. The cell may be a cancer cell. The cancer cell may be of any cancer type described herein. The cell may be in vivo or in vitro.

COMBINATION THERAPY

[00192] Methods of the invention may include a compound of the invention used alone or in combination with one or more additional therapies (e.g., non-drug treatments or therapeutic agents). Dosages of one or more of the additional therapies (e.g., non-drug treatments or therapeutic agents) may be reduced from standard dosages when administered alone. For example, doses may be determined empirically from combinations and permutations of drugs or may be deduced by isobolographic analysis (e.g., Black et al., Neurology 65:S3-S6 (2005)).

[00193] A compound of the present invention may be administered prior to, following, or simultaneously with one or more such additional therapies. When combined, dosages of a compound of the invention and dosages of one or more additional therapies (e.g., non-drug treatment or therapeutic agent) provide a therapeutic effect (e.g., synergistic or additive therapeutic effect). A compound of the present invention and an additional therapy, such as an anticancer agent, may be administered together, such as in a unitary pharmaceutical composition, or separately, and when administered separately, this may occur simultaneously or sequentially. Such sequential administration may be close or remote in time.

[00194] In some embodiments, additional therapy is the administration of side effect limiting agents (e.g., agents intended to decrease the occurrence or severity of side effects of treatment. For example, in some embodiments, the compounds of the present invention can also be used in combination with a therapeutic agent that treats nausea. Examples of agents that can be used to treat nausea include: dronabinol, granisetron, metoclopramide, ondansetron, and prochlorperazine, or pharmaceutically acceptable salts thereof.

[00195] In some embodiments, the one or more additional therapies include a non-drug treatment (e.g., surgery or radiation therapy). In some embodiments, the one or more additional therapies include a therapeutic agent (e.g., a compound or biologic that is an anti-angiogenic agent, signal transduction inhibitor, anti-proliferative agent, glycolysis inhibitor, or autophagy inhibitor). In some embodiments, the one or more additional therapies include a non-drug treatment (e.g., surgery or radiation therapy) and a therapeutic agent (e.g., a compound or biologic that is an anti-angiogenic agent, signal transduction inhibitor, anti-proliferative agent, glycolysis inhibitor, or autophagy inhibitor). In other embodiments, the one or more additional therapies include two therapeutic agents. In still other embodiments, the one or more additional therapies include three therapeutic agents. In some embodiments, the one or more additional therapies include four or more therapeutic agents.

[00196] In this Combination Therapy section, all references are incorporated by reference to the agents described, whether explicitly stated as such or not.

NON-DRUG THERAPIES

[00197] Examples of non-drug treatments include, but are not limited to, radiation therapy, cryotherapy, hyperthermia, surgery (e.g., surgical excision of tumor tissue), and adoptive T-cell transfer therapy (ACT).

[00198] In some embodiments, the compounds of the invention can be used as an adjuvant therapy following surgery. In some embodiments, the compounds of the invention can be used as a neoadjuvant therapy prior to surgery.

[00199] Radiation therapy can be used to inhibit abnormal cell growth or treat a hyperproliferative disorder, such as cancer, in a subject (e.g., mammal (e.g., human)). Techniques for administering radiation therapy are known in the art. Radiation therapy can be administered by one of several methods, or a combination of methods, including, without limitation, external beam therapy, internal radiation therapy, implant radiation, stereotactic radiosurgery, systemic radiation therapy, permanent or temporary interstitial radiation therapy and brachytherapy. The term “brachytherapy,” as used herein, refers to radiation therapy delivered by a spatially confined radioactive material inserted into the body at or near a tumor or other site of proliferative tissue disease. The term is intended, without limitation, to include exposure to radioactive isotopes (e.g., At-211, I-131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32, and radioactive isotopes of Lu). Suitable radiation sources for use as a cell conditioner of the present invention include both solids and liquids. By way of non-limiting example, the radiation source may be a radionuclide such as I-125, I-131, Yb-169, Ir-192 as a solid source, I-125 as a solid source, or other radionuclides that emit photons, beta particles, gamma rays, α-rays, or other therapeutic rays. The radioactive material may also be a fluid made from any radionuclide solution, for example, a solution of I-125 or I-131, or a radioactive fluid may be produced using a suitable fluid substance containing small solid radionuclide particles, such as Au-198, or Y-90. In addition, the radionuclide(s) may be incorporated in the form of radioactive gel or microspheres.

[00200] In some embodiments, the compounds of the present invention can make abnormal cells more sensitive to radiation treatment for the purpose of killing or inhibiting the growth of such cells. Thus, this invention further relates to a method for sensitizing abnormal cells in a mammal to radiation treatment comprising administering to the mammal an amount of a compound of the present invention, which amount is effective in sensitizing abnormal cells to radiation treatment. The amount of the compound in this method can be determined according to the means for determining effective amounts of such compounds described herein. In some embodiments, the compounds of the present invention can be used as an adjuvant therapy following radiation therapy or as a neoadjuvant therapy prior to radiation therapy.

[00201] In some embodiments, the non-pharmacological treatment is an adoptive T cell transfer (ACT) therapy. In some embodiments, the T cell is an activated T cell. The T cell may be modified to express a chimeric antigen receptor (CAR). CAR-modified T cells (CAR-T) may be generated by any method known in the art. For example, CAR-T cells may be generated by introducing a suitable expression vector encoding the CAR into a T cell. Prior to expansion and genetic modification of the T cells, a source of T cells is obtained from a subject. T cells may be obtained from a variety of sources, including peripheral blood mononuclear cells, bone marrow, lymph node tissue, umbilical cord blood, thymus tissue, tissue from a site of infection, ascites, pleural effusion, spleen tissue, and tumors. In certain embodiments of the present invention, any number of T cell lines available in the art may be used. In some embodiments, the T cell is an autologous T cell. Whether before or after genetic modification of T cells to express a desirable protein (e.g., a CAR), T cells can be activated and expanded generally using methods as described, for example, in U.S. Patents 6,352,694; 6,534,055; 6,905,680; 6,692,964; 5,858,358; 6,887,466; 6,905,681; 7,144,575; 7,067,318; 7,172,869; 7,232,566; 7,175,843; 7,572,631; 5,883,223; 6,905,874; 6,797,514; and 6,867,041.

THERAPEUTIC AGENTS

[00202] A therapeutic agent may be a compound used in the treatment of cancer or symptoms associated with it.

[00203] For example, a therapeutic agent may be a steroid. Accordingly, in some embodiments, one or more additional therapies include a steroid. Suitable steroids may include, but are not limited to, 21-acetoxypregnenolone, alclomethasone, algestone, amcinonide, beclomethasone, betamethasone, budesonide, chlorprednisone, clobetasol, clocortolone, cloprednol, corticosterone, cortisone, cortivazole, deflazacort, desonide, desoximethasone, dexamethasone, diflorasone, diflucortolone, difuprednate, enoxolone, fluazacort, flucloronide, flumethasone, flunisolide, fluocinolone acetonide, fluocinonide, butylfluocortin, fluocortolone, fluorometholone, fluperolone acetate, fluprednidene acetate, fluprednisolone, flurandrenolide, fluticasone propionate, formocortal, halcinonide, halobetasol propionate, halomethasone, hydrocortisone, loteprednol etabonate, mazipredone, medrysone, meprednisone, methylprednisolone, mometasone furoate, paramethasone, prednicarbate, prednisolone, 25-diethylaminoacetate, prednisolone sodium phosphate, prednisone, prednival, prednylidene, rimexolone, tixocortol, triamcinolone, triamcinolone acetonide, triamcinolone benetonide, triamcinolone hexacetonide and salts or derivatives thereof.

[00204] Other examples of therapeutic agents that may be used in combination therapy with a compound of the present invention include compounds described in the following patents: U.S. Patents 6,258,812 , 6,630,500 , 6,515,004 , 6,713,485 , 5,521,184 , 5,770,599 , 5,747,498 , 5,990,141 , 6,235,764 , and 8,623,885 , and International Patent Applications WO01/37820 , WO01/32651 , WO02/68406 , WO02/66470 , WO02/55501 , WO04/05279 , WO04/07481 , WO04/07458 , WO04/09784, WO02/59110, WO99/45009, WO00/59509, WO99/61422, WO00/12089 and WO00/02871.

[00205] A therapeutic agent can be a biologic (e.g., cytokine (e.g., interferon or an interleukin, such as IL-2)) used in the treatment of cancer or symptoms associated therewith. In some embodiments, the biologic is an immunoglobulin-based biologic, e.g., a monoclonal antibody (e.g., a humanized antibody, a fully human antibody, an Fc fusion protein, or a functional fragment thereof) that agonizes a target to stimulate an anti-cancer response or antagonizes an antigen important for cancer. Also included are antibody-drug conjugates.

[00206] A therapeutic agent can be a T-cell checkpoint inhibitor. In one embodiment, the checkpoint inhibitor is an inhibitory antibody (e.g., a monospecific antibody, such as a monoclonal antibody). The antibody can be, for example, humanized or fully human. In some embodiments, the checkpoint inhibitor is a fusion protein, for example, an Fc receptor fusion protein. In some embodiments, the checkpoint inhibitor is an agent, such as an antibody, that interacts with a checkpoint protein. In some embodiments, the checkpoint inhibitor is an agent, such as an antibody, that interacts with the ligand of a checkpoint protein. In some embodiments, the checkpoint inhibitor is an inhibitor (e.g., an inhibitory antibody or small molecule inhibitor) of CTLA-4 (e.g., an anti-CTLA-4 antibody or a fusion protein). In some embodiments, the checkpoint inhibitor is an inhibitor or antagonist (e.g., an inhibitory antibody or small molecule inhibitor) of PD-1. In some embodiments, the checkpoint inhibitor is an inhibitor or antagonist (e.g., an inhibitory antibody or small molecule inhibitor) of PDL-1. In some embodiments, the checkpoint inhibitor is an inhibitor or antagonist (e.g., an inhibitory antibody or Fc fusion or small molecule inhibitor) of PDL-2 (e.g., a PDL-2/Ig fusion protein). In some embodiments, the checkpoint inhibitor is an inhibitor or antagonist (e.g., an inhibitory antibody or small molecule inhibitor) of B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, vista, KIR, 2B4, CD160, CGEN-15049, CHK 1, CHK2, A2aR, B-7 family ligands, or a combination thereof. In some embodiments, the checkpoint inhibitor is pembrolizumab, nivolumab, PDR001 (NVS), REGN2810 (Sanofi/Regeneron), a PD-L1 antibody such as, e.g., avelumab, durvalumab, atezolizumab, pidilizumab, JNJ-63723283 (JNJ), BGB-A317 (BeiGene & Celgene), or a checkpoint inhibitor disclosed in Preusser, M. et al. (2015) Nat. Rev. Neurol., including but not limited to ipilimumab, tremelimumab, nivolumab, pembrolizumab, AMP224, AMP514/MEDI0680, BMS936559, MEDl4736, MPDL3280A,MSB0010718C, BMS986016, IMP321, lirilumab, IPH2101, 1-7F9, and KW-6002.

[00207] A therapeutic agent may be an anti-TIGIT antibody, such as MBSA43, BMS-986207, MK-7684, COM902, AB154, MTIG7192A, or OMP-313M32 (etigilimab).

[00208] A therapeutic agent may be an agent that treats cancer or symptoms associated therewith (e.g., a cytotoxic agent, non-peptide small molecule, or other compound useful in treating cancer or symptoms associated therewith, collectively, an “anti-cancer agent”). Anti-cancer agents may be, for example, chemotherapeutic or targeted therapy agents.

[00209] Anticancer agents include mitotic inhibitors, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, alkylating agents, antimetabolites, folic acid analogues, pyrimidine analogues, purine analogues and related inhibitors, vinca alkaloids, epipodopylotoxins, antibiotics, L-asparaginase, topoisomerase inhibitors, interferons, platinum coordination complexes, anthracenedione-substituted urea, methylhydrazine derivatives, adrenocortical suppressant, adrenocorticosteroids, progestins, estrogens, antiestrogens, androgens, antiandrogens, and gonadotropin-releasing hormone analogues. Other anticancer agents include leucovorin (LV), irenotecan, oxaliplatin, capecitabine, paclitaxel, and doxetaxel. In some embodiments, the one or more additional therapies include two or more anticancer agents. The two or more anticancer agents can be used in a cocktail to be administered in combination or administered separately. Suitable dosing regimens of combination anticancer agents are known in the art and described in, for example, Saltz et al., Proc. Am. Soc. Clin. Oncol. 18:233a (1999), and Douillard et al., Lancet 355(9209):1041-1047 (2000).

[00210] Other non-limiting examples of anticancer agents include Gleevec® (Imatinib Mesylate); Kyprolis® (carfilzomib); Velcade® (bortezomib); Casodex (bicalutamide); Iressa® (gefitinib); alkylating agents such as thiotepa and cyclophosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethylolomelamine; acetogenins (especially bulatacin and bulatacinone); a camptothecin (including the synthetic analogue topotecan); bryostatin; kallistatin; CC-1065 (including its synthetic analogues adozelesin, carzelesin, and bizelesin); cryptophycins (particularly cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including the synthetic analogues, KW-2189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictin A; spongistatin; nitrogen mustards such as chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembiquin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimustine; antibiotics, such as the enediine antibiotics (e.g., calicheamicin, such as gamal calicheamicin and omegal calicheamicin (see, e.g., Agnew, Chem. Intl. Ed Engl. 33:183-186 (1994)); dynemycin, such as dynemycin A; bisphosphonates, such as clodronate; a esperamycin; neocarzinostatin chromophore and chromoprotein related, chromophores antibiotics enediin, aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, caqueamicin, carabicin, caminomycin, carminomycin, carzinophylline, chromomycins, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, adriamycin (doxorubicin), morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, desoxydoxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, chelamycin, rhodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, pteropterin, trimetrexate; purine analogues such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogues such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; antiadrenals such as aminoglutethimide, mitotane, trilostane; folic acid replenishers such as phrolininic acid; aceglaton; aldophosphamide glucoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; desofamine; demecolcine; diaziquone; elfomitin; elliptin acetate; an epothilone such as epothilone B; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidamine; maytansinoids, such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; fenamet; pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK® polysaccharide complex (JHS Natural Products, Eugene, OR); razoxane; rhizoxin; sizofiran; spirogermanium; tenuazonic acid; triziquone; 2,2',2''-trichlorotriethylamine; trichothecenes, such as T-2 toxin, verracurin A, roridine A, and anguidin; urethane; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobromane; gacytosine; arabinoside (“Ara-C”); cyclophosphamide; thiotepa; taxoids, e.g., Taxol® (paclitaxel), Abraxane® (cremophor-free albumin-engineered paclitaxel nanoparticle formulation), and Taxotere® (doxetaxel); chlorambucil; tamoxifen (Nolvadex™); raloxifene; 4(5)-imidazole aromatase inhibitors; 4-hydroxytamoxifen; trioxifen; ceoxifen; LY 117018; onapristone; toremifene (Fareston®); flutamide, nilutamide, bicalutamide, leuprolide, goserelin; chlorambucil; gemcitabine Gemzar®; 6-thioguanine; mercaptopurine; platinum coordination complexes such as cisplatin, oxaliplatin, and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; Navelbine® (vinorelbine); novatrone; teniposide; edatrexate; daunomycin; aminopterin; ibandronate; irrinotecan (e.g., CPT-11); topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids, such as retinoic acid; esperamycins; capecitabine (e.g., Xeloda®); and pharmaceutically acceptable salts of any of the foregoing.

[00211] Additional non-limiting examples of anticancer agents include trastuzumab (Herceptin®), bevacizumab (Avastin®), cetuximab (Erbitux®), rituximab (Rituxan®), Taxol®, Arimidex®, ABVD, avicin, abagovomab, acridine carboxamide, adecatumumab, 17-N-allylamino-17-demethoxygeldanamycin, alfaradin, alvocidib, 3-aminopyridine-2-carboxaldehyde thiosemicarbazone, amonafide, anthracenedione, anti-CD22 immunotoxins, antineoplastics (e.g., cell cycle nonspecific antineoplastic agents and other antineoplastics described herein), antitumorigenic herbs, apaziquone, atiprimod, azathioprine, belotecan, bendamustine, BIBW 2992, biricodar, brostallicin, bryostatin, buthionine sulfoximine, CBV (chemotherapy), calyculin, dichloroacetic acid, discodermolide, elsamitrucin, enocitabine, eribulin, exatecan, exisulinde, ferruginol, forodesine, phosphetrol, ICE chemotherapy regimen, IT-101, imexon, imiquimod, indolocarbazole, irofulvene, laniquidar, larotaxel, lenalidomide, lucanthone, lurtotecan, mafosfamide, mitozolomide, nafoxidine, nedaplatin, olaparib, ortataxel, PAC-1, pawpaw, pixantrone, proteasome inhibitors, rebeccamycin, resiquimod, rubitecan, SN-38, salinosporamide A, sapacitabine, Stanford V, swainsonine, talaporfin, tariquidar, tegafur-uracil, temodar,thesistaxel, triplatin tetranitrate, tris(2-chloroethyl)amine, troxacitabine, uramustine, vadimezan, vinflunine, ZD6126 and zosuquidar.

[00212] Other non-limiting examples of anticancer agents include natural products such as vinca alkaloids (e.g., vinblastine, vincristine, and vinorelbine), epidipodophyllotoxins (e.g., etoposide and teniposide), antibiotics (e.g., dactinomycin (actinomycin D), daunorubicin, and idarubicin), anthracyclines, mitoxantrone, bleomycins, plicamycin (mithramycin), mitomycin, enzymes (e.g., L-asparaginase which systemically metabolizes L-asparagine and deprives cells that lack the ability to synthesize their own asparagine), antiplatelet agents, antiproliferative/antimitotic alkylating agents such as nitrogen mustards (e.g., mechlorethamine, cyclophosphamide and analogues, melphalan, and chlorambucil), ethylenimines, and methylmelamines (e.g., hexamethylmelamine and thiotepa), CDK inhibitors (e.g., a CDK4/6 inhibitor such as abemaciclib, ribociclib, palbociclib; seliciclib, UCN-01, P1446A-05, PD-0332991, dinaciclib, P27-00, AT-7519, RGB286638, and SCH727965), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine (BCNU) and analogues, and streptozocin), trazodone-dacarbazinine (DTIC), antiproliferative/antimitotic antimetabolites such as folic acid analogues, pyrimidine analogues (e.g., fluorouracil, floxuridine, and cytarabine), purine analogues and related inhibitors (e.g., mercaptopurine, thioguanine, pentostatin, and 2-chlorodeoxyadenosine), aromatase inhibitors (e.g., anastrozole, exemestane, and letrozole) and platinum coordination complexes (e.g., cisplatin and carboplatin), procarbazine, hydroxyurea, mitotane, aminoglutethimide, histone deacetylase (HDAC) inhibitors (e.g., trichostatin, sodium butyrate, apicidan, suberoyl anilide hydroamic acid, vorinostat, LBH 589, romidepsin, ACY-1215, and panobinostat), mTOR inhibitors (e.g., vistusertib, temsirolimus, everolimus, ridaforolimus, and sirolimus), KSP(Eg5) inhibitors (e.g., Array 520), DNA binding agents (e.g., Zalypsis®), PI3K inhibitors such as such as PI3K delta inhibitor (e.g., GS-1101 and TGR-1202), PI3K delta and gamma inhibitor (e.g., CAL-130), copanlisib, alpelisib, and idelalisib; multikinase inhibitor (e.g., TG02 and sorafenib), hormones (e.g., estrogen) and hormonal agonists such as leutinizing hormone-releasing hormone (LHRH) agonists (e.g., goserelin, leuprolide, and triptorelin), BAFF neutralizing antibody (e.g., LY2127399), IKK inhibitors, p38MAPK inhibitors, anti-IL-6 (e.g., CNT0328), telomerase inhibitors (e.g., GRN 163L), aurora kinase inhibitors (e.g., MLN8237), cell surface monoclonal antibodies (e.g., anti-CD38 (HUMAX-CD38), anti-CSl (e.g., elotuzumab), HSP90 inhibitors (e.g., 17 AAG and KOS 953), P13K/Akt inhibitors (e.g., perifosine), Akt inhibitors (e.g., GSK-2141795), PKC inhibitors (e.g., enzastaurin), FTIs (e.g., Zarnestra™), anti-CD138 (e.g., BT062), Torcl/2-specific kinase inhibitors (e.g., INK128), ER/UPR targeting agents (e.g., MKC-3946), cFMS inhibitors (e.g., ARRY-382), JAK1/2 inhibitors (e.g., CYT387), PARP inhibitors (e.g., olaparib and veliparib (ABT-888)), and BCL-2 antagonists.

[00213] In some embodiments, an anticancer agent is selected from mechlorethamine, camptothecin, ifosfamide, tamoxifen, raloxifene, gemcitabine, Navelbine®, sorafenib, or any analog or variant derived from the foregoing.

[00214] In some embodiments, the anticancer agent is a HER2 inhibitor. Non-limiting examples of HER2 inhibitors include monoclonal antibodies such as trastuzumab (Herceptin®) and pertuzumab (Perjeta®); small molecule tyrosine kinase inhibitors such as gefitinib (Iressa®), erlotinib (Tarceva®), pilitinib, CP-654577, CP-724714, canertinib (CI 1033), HKI-272, lapatinib (GW-572016; Tykerb®), PKI-166, AEE788, BMS-599626, HKI-357, BIBW 2992, ARRY-334543, and JNJ-26483327.

[00215] In some embodiments, an anticancer agent is an ALK inhibitor. Non-limiting examples of ALK inhibitors include ceritinib, TAE-684 (NVP-TAE694), PF02341066 (crizotinib or 1066), alectinib; brigatinib; entrectinib; ensartinib (X-396); lorlatinib; ASP3026; CEP-37440; 4SC-203; TL-398; PLB1003; TSR-011; CT-707; TPX-0005, and AP26113. Additional examples of ALK kinase inhibitors are described in Examples 3-39 of WO05016894.

[00216] In some embodiments, an anti-cancer agent is an inhibitor of a downstream member of a Receptor Tyrosine Kinase (RTK)/Growth Factor Receptor (e.g., a SHP2 inhibitor (e.g., SHP099, TNO155, RMC-4550, RMC-4630, JAB-3068, RLY-1971), a SOS1 inhibitor (e.g., BI-1701963, BI-3406), a Raf inhibitor, a MEK inhibitor, an ERK inhibitor, a PI3K inhibitor, a PTEN inhibitor, an AKT inhibitor, or an mTOR inhibitor (e.g., mTORC1 inhibitor or mTORC2 inhibitor). In some embodiments, the anti-cancer agent is JAB-3312. In some embodiments, an anti-cancer agent is an additional Ras inhibitor (e.g., AMG 510, MRTX1257, MRTX849, JNJ-74699157 (ARS-3248), LY3499446, ARS-853 or ARS-1620), or a Ras vaccine, or other therapeutic modality designed to directly or indirectly decrease the oncogenic activity of Ras. Other examples of Ras inhibitors that can be combined with a Ras inhibitor of the present invention are provided below, incorporated herein by reference in their entirety: WO 2020050890, WO 2020047192, WO 2020035031, WO 2020028706, WO2019241157, WO 2019232419, WO 2019217691, WO 2019217307, WO 2019215203, WO 2019213526, WO2019213516, WO2019155399, WO2019150305, WO2019110751, WO2019099524, WO2019051291, WO2018218070, WO 2018217651, WO 2018218071, WO 2018218069, WO2018206539, WO 2018143315, WO 2018140600, WO 2018140599, WO2018140598, WO 2018140514, WO 2018140513, WO 2018140512, WO2018119183, WO 2018112420, WO 2018068017, WO 2018064510, WO2017201161, WO 2017172979, WO 2017100546, WO 2017087528, WO2017058807, WO 2017058805, WO 2017058728, WO 2017058902, WO2017058792, WO 2017058768, WO 2017058915, WO 2017015562, WO2016168540, WO 2016164675, WO 2016049568, WO 2016049524, WO2015054572, WO 2014152588, WO 2014143659 and WO 2013155223.

[00217] In some embodiments, a therapeutic agent that can be combined with a compound of the present invention is a MAP kinase (MAPK) pathway inhibitor (or “MAPK inhibitor”). MAPK inhibitors include, but are not limited to, one or more MAPK inhibitors described in Cancers (Basel) 2015 Sep; 7(3): 1758-1784. For example, the MAPK inhibitor can be selected from one or more of trametinib, binimetinib, selumetinib, cobimetinib, LErafAON (NeoPharm), ISIS 5132; vemurafenib, pimasertib, TAK733, RO4987655 (CH4987655); CI-1040; PD-0325901; CH5126766; MAP855; AZD6244; refametinib (RDEA 119/BAY 86-9766); GDC- 0973/XL581; AZD8330 (ARRY-424704/ARRY-704); RO5126766 (Roche, described in PLoS One. 2014 Nov 25;9(11)); and GSK1120212 (or JTP-74057, described in Clin Cancer Res. 2011 Mar 1;17(5):989-1000). The MAPK inhibitor could be PLX8394, LXH254, GDC-5573 or LY3009120.

[00218] In some embodiments, an anti-cancer agent is a disruptor or inhibitor of the RAS-RAF-ERK or PI3K-AKT-TOR or PI3K-AKT signaling pathways. The PI3K/AKT inhibitor may include, but is not limited to, one or more PI3K/AKT inhibitors described in Cancers (Basel) 2015 Sep; 7(3): 1758-1784. For example, the PI3K/AKT inhibitor may be selected from one or more of NVP-BEZ235; BGT226; XL765/SAR245409; SF1126; GDC-0980; PI-103; PF-04691502; PKI-587; GSK2126458.

[00219] In some embodiments, an anti-cancer agent is a PD-1 or PD-L1 antagonist.

[00220] In some embodiments, additional therapeutic agents include ALK inhibitors, HER2 inhibitors, EGFR inhibitors, IGF-1R inhibitors, MEK inhibitors, PI3K inhibitors, AKT inhibitors, TOR inhibitors, MCL-1 inhibitors, BCL-2 inhibitors, SHP2 inhibitors, proteasome inhibitors, and immune therapies. In some embodiments, a therapeutic agent can be a pan-RTK inhibitor, such as afatinib.

[00221] IGF-1R inhibitors include linsitinib, or a pharmaceutically acceptable salt thereof.

[00222] EGFR inhibitors include, but are not limited to, small molecule antagonists, antibody inhibitors, or specific antisense nucleotide or siRNA. Useful antibody inhibitors of EGFR include cetuximab (Erbitux®), panitumumab (Vectibix®), zalutumumab, nimotuzumab, and matuzumab. Other antibody-based EGFR inhibitors include any anti-EGFR antibody or antibody fragment that can partially or completely block the activation of EGFR by its natural ligand. Non-limiting examples of antibody-based EGFR inhibitors include those described in Modjtahedi et al., Br. J. Cancer 1993, 67:247-253; Teramoto et al., Cancer 1996, 77:639-645; Goldstein et al., Clin. Cancer Res. 1995, 1:1311-1318; Huang et al., 1999, Cancer Res. 15:59(8):1935-40; and Yang et al., Cancer Res. 1999, 59:1236-1243. The EGFR inhibitor may be the monoclonal antibody Mab E7.6.3 (Yang, 1999 supra), or Mab C225 (ATCC Accession No. HB-8508), or an antibody or antibody fragment with the same binding specificity.

[00223] Small molecule EGFR antagonists include gefitinib (Iressa®), erlotinib (Tarceva®), and lapatinib (TykerB®). See, for example, Yan et al., Pharmacogenetics and Pharmacogenomics In Oncology Therapeutic Antibody Development, BioTechniques 2005, 39(4):565-8; and Paez et al., EGFR Mutations In Lung Cancer Correlation With Clinical Response To Gefitinib Therapy, Science 2004, 304(5676):1497-500. In some embodiments, the EGFR inhibitor is osimertinib (Tagrisso®). Other non-limiting examples of small molecule EGFR inhibitors include any of the EGFR inhibitors described in the following patent publications and all pharmaceutically acceptable salts of such EGFR inhibitors: EP 0520,722; EP 0566,226; WO96/33980; U.S. Pat. 5,747,498; WO96/30347; EP0787772; WO97/30034; WO97/30044; WO97/38994; WO97/49688; EP 837063; WO98/02434; WO97/38983; WO95/19774; WO95/19970; WO97/13771; WO98/02437; WO98/02438; WO97/32881; DE 19629652; WO98/33798; WO97/32880; WO97/32880; EP 682027; WO97/02266; WO97/27199; WO98/07726; WO97/34895; WO96/31510; WO98/14449; WO98/14450;WO98/14451; WO95/09847; WO97/19065; WO98/17662; Pat. U.S. 5,789,427; Pat. U.S. 5,650,415; Pat. U.S. 5,656,643; WO99/35146; WO99/35132; WO99/07701; and WO92/20642. Additional non-limiting examples of small molecule EGFR inhibitors include any of the EGFR inhibitors described in Traxler et al., Exp. Opin. Ther. Patents 1998, 8(12):1599-1625. In some embodiments, an EGFR inhibitor is an ERBB inhibitor. In humans, the ERBB family contains HER1 (EGFR, ERBB1), HER2 (NEU, ERBB2), HER3 (ERBB3), and HER (ERBB4).

[00224] MEK inhibitors include, but are not limited to, pimasertib, selumetinib, cobimetinib (Cotellic®), trametinib (Mekinist®), and binimetinib (Mektovi®). In some embodiments, a MEK inhibitor targets a MEK mutation that is a Class I MEK1 mutation selected from D67N; P124L; P124S; and L177V. In some embodiments, the MEK mutation is a Class II MEK1 mutation selected from ΔE51-Q58; ΔF53-Q58; E203K; L177M; C121S; F53L; K57E; Q56P; and K57N.

[00225] PI3K inhibitors include, but are not limited to, wortmannin; 17-hydroxywortmannin analogues described in WO06/044453; 4-[2-(1H-Indazol-4-yl)-6-[[4-(methylsulfonyl)piperazin-1-yl]methyl]thieno[3,2-d]pyrimidin-4-yl]morpholine (also known as pictilisib or GDC-0941 and described in WO09/036082 and WO09/055730); 2-methyl-2-[4-[3-methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydroimidazo[4,5-c]quinolin-1-yl]phenyl]propionitrile (also known as BEZ 235 or NVP-BEZ 235, and described in WO06/122806); (S)-1-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2-hydroxypropan-1-one (described in WO08/070740); LY294002 (2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (available from Axon Medchem); PI 103 hydrochloride (3-[4-(4-morpholinylpyrido-[3',2':4,5]furo[3,2-d]pyrimidin-2-yl]phenol hydrochloride (available from Axon Medchem); PIK 75 (2-methyl-5-nitro-2-[(6-bromoimidazo[1,2-a]pyridin-3-yl)methylene]-1-methylhydrazide-benzenesulfonic acid monohydrochloride) (available from Axon Medchem); PIK 90 (N-(7,8-dimethoxy-2,3-dihydro-imidazo[1,2-c]quinazolin-5-yl)-nicotinamide (available from Axon Medchem); AS-252424 (5-[l-[5-(4-fluoro-2-hydroxy-phenyl)-furan-2-yl]-meth-(Z)-ylidene]-thiazolidine-2,4-dione (available from Axon Medchem); TGX-221 (7 -methyl-2-(4-morpholinyl)-9-[1-(phenylamino)ethyl]-4H-pyrido-[1,2-a]pyrimidin-4-one (available from Axon Medchem); INK1117, IPI-145, BKM120, XL147, XL765, Palomid 529, GSK1059615, ZSTK474, PWT33597, IC87114, TGI 00-115, CAL263, PI-103, GNE-477, CUDC-907, and AEZS-136.

[00226] AKT inhibitors include, but are not limited to, Akt-1-1 (inhibits Aktl) (Barnett et al., Biochem. J. 2005, 385(Pt. 2): 399-408); Akt- 1-1,2 (inhibits Akl e 2) (Barnett et al., Biochem. J. 2005, 385(Pt. 2): 399-408); API-59CJ-Ome (e.g., Jin et al., Br. J. Cancer 2004, 91:1808-12); 1-H-imidazo[4,5-c]pyridinyl compounds (e.g., WO 05/011700); indole-3-carbinol and derivatives thereof (e.g., U.S. Pat. 6,656,963; Sarkar and Li J Nutr. 2004, 134(12 Suppl): 3493S-3498S); perifosine (e.g., interferes with Akt membrane localization; Dasmahapatra et al. Clin. Cancer Res. 2004, 10(15): 5242-52); phosphatidylinositol ether lipid analogs (e.g., Gills and Dennis Expert. Opin. Investig. Drugs 2004, 13:787-97); and triciribine (TCN or API-2 or NCI identifier: NSC 154020; Yang et al., Cancer Res. 2004, 64:4394-9).

[00227] mTOR inhibitors include, but are not limited to, ATP-competitive mTORC1/mTORC2 inhibitors, e.g., PI-103, PP242, PP30; Torin 1; FKBP12 enhancers; 4H-1-benzopyran-4-one derivatives; and rapamycin (also known as sirolimus) and derivatives thereof, including: temsirolimus (Torisel®); everolimus (Afinitor®; WO94/09010); ridaforolimus (also known as deforolimus or AP23573); rapalogs, e.g., as disclosed in WO98/02441 and WO01/14387, e.g., AP23464 and AP23841; 40-(2-hydroxyethyl)rapamycin; 40-[3-hydroxy(hydroxymethyl)methylpropanoate]-rapamycin (also known as CC1779); 40-epi-(tetrazolit)-rapamycin (also called ABT578); 32-deoxorapamycin; 16-pentynyloxy-32(S)-dihydrorapamycin; derivatives disclosed in WO05/005434; derivatives disclosed in U.S. Patents 5,258,389, 5,118,677, 5,118,678, 5,100,883, 5,151,413, 5,120,842, and 5,256,790, and in WO94/090101, WO92/05179, WO93/111130, WO94/02136, WO94/02485, WO95/14023, WO94/02136, WO95/16691, WO96/41807, WO96/41807, and WO2018204416; and phosphorus-containing rapamycin derivatives (e.g., WO05/016252). In some embodiments, the mTOR inhibitor is a bisteric inhibitor (see, e.g., WO2018204416, WO2019212990, and WO2019212991), such as RMC-5552.

[00228] BRAF inhibitors that can be used in combination with the compounds of the invention include, for example, vemurafenib, dabrafenib, and encorafenib. A BRAF can comprise a Class 3 BRAF mutation. In some embodiments, the Class 3 BRAF mutation is selected from one or more of the following amino acid substitutions in human BRAF: D287H; P367R; V459L; G466V; G466E; G466A; S467L; G469E; N581S; N581I; D594N; D594G; D594A; D594H; F595L; G596D; G596R, and A762E.

[00229] MCL-1 inhibitors include, but are not limited to, AMG-176, MIK665, and S63845. Myeloid cell leukemia-1 (MCL-1) protein is a major anti-apoptotic member of the B-cell lymphoma-2 (BCL-2) family of proteins. Overexpression of MCL-1 has been closely linked to tumor progression as well as resistance to not only traditional chemotherapies but also targeted therapeutics including BCL-2 inhibitors such as ABT-263.

[00230] In some embodiments, the additional therapeutic agent is an inhibitor of SHP2. SHP2 is a non-receptor protein tyrosine phosphatase encoded by the PTPN11 gene that contributes to multiple cellular functions, including proliferation, differentiation, cell cycle maintenance, and migration. SHP2 has two N-terminal Src homology 2 domains (N-SH2 and C-SH2), a catalytic domain (PTP), and a C-terminal tail. The two SH2 domains control the subcellular localization and functional regulation of SHP2. The molecule exists in an inactive, autoinhibited conformation stabilized by a binding network involving residues from the N-SH2 and PTP domains. Stimulation by, for example, cytokines or growth factors that act through receptor tyrosine kinases (RTKs) leads to exposure of the catalytic site, resulting in enzymatic activation of SHP2.

[00231] SHP2 is involved in signaling through the RAS-mitogen-activated protein kinase (MAPK), JAK-STAT, or phosphoinositol 3-kinase-AKT pathways. Mutations in the PTPN11 gene and subsequently in SHP2 have been identified in several human developmental diseases, such as Noonan syndrome and Leopard syndrome, as well as in human cancers, such as juvenile myelomonocytic leukemia, neuroblastoma, melanoma, acute myeloid leukemia, and breast, lung, and colon cancer. Some of these mutations destabilize the autoinhibited conformation of SHP2 and promote autoactivation or growth factor-enhanced activation of SHP2. SHP2 therefore represents a highly attractive target for the development of novel therapies for the treatment of several diseases, including cancer. A SHP2 inhibitor (e.g., RMC-4550 or SHP099) in combination with a RAS pathway inhibitor (e.g., a MEK inhibitor) has been shown to inhibit the proliferation of multiple cancer cell lines in vitro (e.g., pancreatic, lung, ovarian, and breast cancer). Thus, combination therapy involving a SHP2 inhibitor with a RAS pathway inhibitor may be a general strategy to prevent tumor resistance in a broad range of malignancies.

[00232] Non-limiting examples of such SHP2 inhibitors that are known in the art include: Chen et al. Mol Pharmacol. 2006, 70, 562; Sarver et al., J. Med. Chem. 2017, 62, 1793; Xie et al., J. Med. Chem. 2017, 60, 113734; and gbe et al., Oncotarget, 2017, 8, 113734; and PCT applications: WO2015107493; WO2015107494; WO201507495; WO2016203404; WO2016203405; WO2016203406; WO2011022440; WO2017156397;WO2017079723; WO2017211303; WO2012041524; WO2017211303;WO2019051084; WO2017211303; US20160030594; US20110281942;WO2010011666; WO2014113584; WO2014176488; WO2017100279;WO2019051469; US8637684; WO2007117699; WO2015003094; WO2005094314; WO2008124815; WO2009049098; WO2009135000; WO2016191328; WO2016196591; WO2017078499; WO2017210134; WO2018013597; WO2018129402; WO2018130928; WO20181309928; WO2018136264; WO2018136265; WO2018160731; WO2018172984; and WO2010121212, each of which is incorporated by reference.

[00233] In some embodiments, a SHP2 inhibitor binds to the active site. In some embodiments, a SHP2 inhibitor is a mixed-type irreversible inhibitor. In some embodiments, a SHP2 inhibitor binds to an allosteric site, e.g., a non-covalent allosteric inhibitor. In some embodiments, a SHP2 inhibitor is a covalent SHP2 inhibitor, such as an inhibitor that targets the cysteine residue (C333) that lies outside the phosphatase active site. In some embodiments, a SHP2 inhibitor is a reversible inhibitor. In some embodiments, a SHP2 inhibitor is an irreversible inhibitor. In some embodiments, the SHP2 inhibitor is SHP099. In some embodiments, the SHP2 inhibitor is TNO155. In some embodiments, the SHP2 inhibitor is RMC-4550. In some embodiments, the SHP2 inhibitor is RMC-4630. In some embodiments, the SHP2 inhibitor is JAB-3068. In some embodiments, the SHP2 inhibitor is RLY-1971.

[00234] In some embodiments, the additional therapeutic agent is selected from the group consisting of a MEK inhibitor, a HER2 inhibitor, a SHP2 inhibitor, a CDK4/6 inhibitor, an mTOR inhibitor, a SOS1 inhibitor, and a PD-L1 inhibitor. In some embodiments, the additional therapeutic agent is selected from the group consisting of a MEK inhibitor, a SHP2 inhibitor, and a PD-L1 inhibitor. See, e.g., Hallin et al., Cancer Discovery, DOI: 10.1158/2159-8290 (October 28, 2019) and Canon et al., Nature, 575:217 (2019). In some embodiments, a Ras inhibitor of the present invention is used in combination with a MEK inhibitor and a SOS1 inhibitor. In some embodiments, a Ras inhibitor of the present invention is used in combination with a PDL-1 inhibitor and a SOS1 inhibitor. In some embodiments, a Ras inhibitor of the present invention is used in combination with a PDL-1 inhibitor and a SHP2 inhibitor. In some embodiments, a Ras inhibitor of the present invention is used in combination with a MEK inhibitor and a SHP2 inhibitor. In some embodiments, the cancer is colorectal cancer and the treatment comprises administering a Ras inhibitor of the present invention in combination with a second or third therapeutic agent.

[00235] Proteasome inhibitors include, but are not limited to, carfilzomib (Kyprolis®), bortezomib (Velcade®), and oprozomib.

[00236] Immune therapies include, but are not limited to, monoclonal antibodies, immunomodulatory imides (IMiDs), GITR agonists, genetically engineered T cells (e.g., CAR-T cells), bispecific antibodies (e.g., BiTEs), and anti-PD-1, anti-PDL-1, anti-CTLA4, anti-LAG1, and anti-OX40 agents).

[00237] Immunomodulatory agents (IMiDs) are a class of immunomodulatory drugs (drugs that adjust immune responses) containing an imide group. The IMiD class includes thalidomide and its analogues (lenalidomide, pomalidomide and apremilast).

[00238] Exemplary anti-PD-1 antibodies and methods for their use are described by Goldberg et al., Blood 2007, 110(1):186-192; Thompson et al., Clin. Cancer Res. 2007, 13(6): 1757-1761; and WO06/121168 A1), as well as described elsewhere herein.

[00239] GITR agonists include, but are not limited to, GITR fusion proteins and anti-GITR antibodies (e.g., bivalent anti-GITR antibodies), such as, a GITR fusion protein described in U.S. Pat. 6,111,090, , U.S. Pat. 8,586,023, WO2010/003118, and WO2011/090754; or an anti-GITR antibody described, for example, in U.S. Pat. 7,025,962, EP 1947,183, U.S. Pat. 7,812,135, U.S. Pat. 8,388,967, U.S. Pat. 8,591,886, U.S. Pat. U.S. 7,618,632, EP 1866339 and WO2011/028683, WO2013/039954, WO05/007190, WO07/133822, WO05/055808, WO99/40196, WO01/03720, WO99/20758, WO06/083289, WO05/115451 and WO2011/051726.

[00240] Another example of a therapeutic agent that may be used in combination with the compounds of the invention is an antiangiogenic agent. Antiangiogenic agents include, but are not limited to, chemical compositions prepared synthetically in vitro, antibodies, antigen-binding regions, radionuclides, and combinations and conjugates thereof. An antiangiogenic agent may be an agonist, antagonist, allosteric modulator, toxin, or more generally, may act to inhibit or stimulate its target (e.g., receptor or enzyme activation or inhibition) and thereby promote cell death or arrest cell growth. In some embodiments, one or more additional therapies include an antiangiogenic agent.

[00241] Antiangiogenic agents may be MMP-2 (matrix metalloproteinase 2) inhibitors, MMP-9 (matrix metalloproteinase 9) inhibitors, and COX-II (cyclooxygenase 11) inhibitors. Non-limiting examples of antiangiogenic agents include rapamycin, temsirolimus (CCI-779), everolimus (RAD001), sorafenib, sunitinib, and bevacizumab. Examples of useful COX-II inhibitors include alecoxib, valdecoxib, and rofecoxib. Examples of useful matrix metalloproteinase inhibitors are described in WO96/33172, WO96/27583, WO98/07697, WO98/03516, WO98/34918, WO98/34915, WO98/33768, WO98/30566, WO90/05719, WO99/52910, WO99/52889, WO99/29667, WO99007675, EP0606046, EP0780386, EP1786785, EP1181017, EP0818442, EP1004578 and US20090012085 and U.S. Patents 5,863,949 and 5,861,510. Preferred MMP-2 and MMP-9 inhibitors are those that have little or no MMP-1 inhibitory activity. Most preferred are those that selectively inhibit MMP-2 or AMP-9 over the other matrix metalloproteinases (i.e., MAP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12, and MMP-13). Some specific examples of MMP inhibitors are AG-3340, RO 32-3555, and RS 13-0830.

[00242] Other exemplary anti-angiogenic agents include KDR (kinase domain receptor) inhibitory agents (e.g., antibodies and antigen-binding regions that specifically bind to the kinase domain receptor), anti-VEGF agents (e.g., antibodies or antigen-binding regions that specifically bind to VEGF (e.g., bevacizumab) or soluble VEGF receptors or a ligand-binding region thereof), such as VEGF-TRAP™ and anti-VEGF receptor agents (e.g., antibodies or antigen-binding regions that specifically bind thereto), EGFR inhibitory agents (e.g., antibodies or antigen-binding regions that specifically bind thereto), such as Vectibix® (panitumumab), erlotinib (Tarceva®), anti-Ang1 and anti-Ang2 agents (e.g., antibodies or antigen-binding regions that specifically bind thereto or to their receptors, for (e.g., Tie2/Tek) and anti-Tie2 kinase inhibitory agents (e.g., antibodies or antigen-binding regions that specifically bind to them). Other antiangiogenic agents include Campath, IL-8, B-FGF, Tek antagonists (US2003/0162712; US6,413,932), anti-TWEAK agents (e.g., antibodies binding specifically or antigen-binding regions, or soluble TWEAK receptor antagonists; see US6,727,225), ADAM distintegrin domain to antagonize integrin binding to its ligands (US 2002/0042368), anti-eph or anti-eph receptor antibodies binding specifically or antigen-binding regions (U.S. Patents 5,981,245; 5,728,813; 5,969,110; 6,596,852; 6,232,447; 6,057,124 and members of the patent family of same) and anti-PDGF-BB antagonists (e.g., specific binding antibodies or antigen-binding regions), as well as antibodies or antigen-binding regions that specifically bind to PDGF-BB ligands and PDGFR kinase inhibitory agents (e.g., antibodies or antigen-binding regions that specifically bind thereto). Additional antiangiogenic agents include: SD-7784 (Pfizer, USA); cilengitide (Merck KGaA, Germany, EPO 0770622); pegaptanib octasodium, (Gilead Sciences, USA); Alfastatin, (BioActa, UK); M-PGA, (Celgene, USA, US 5712291); ilomastat, (Arriva, USA, US5892112); emaxanib, (Pfizer, USA, US 5792783); vatalanib, (Novartis, Switzerland); 2-methoxyestradiol (EntreMed, USA); TLC ELL-12 (Elan, Ireland); anecortave acetate (Alcon, USA); alpha-D148 MAb (Amgen, USA); CEP-7055 (Cephalon, USA); anti-Vn MAb (Crucell, Netherlands), DACanthiangiogenic (ConjuChem, Canada); Angiocidin (InKine Pharmaceutical, USA); KM-2550 (Kyowa Hakko, Japan); SU-0879 (Pfizer, USA); CGP-79787 (Novartis, Switzerland, 09700); Argentad Technology, USA); YRealth (YR); Johnson's angiogenic inhibitor (Johnson, UK); TBC-1635 (Encysive Pharmaceuticals, USA); SC-236 (Pfizer, USA); ABT-567 (Abbott, USA); Metastatin (EntreMed, USA); maspin (Sosei, Japan); 2-methoxyestradiol (Oncology Sciences Corporation, USA); ER-68203-00 (IV AX, USA); BeneFin (Lane Labs, USA); Tz-93 (Tsumura, Japan); TAN-1120 (Takeda, Japan); FR-111142 (Fujisawa, Japan, JP 02233610); platelet factor 4 (RepliGen, USA, EP 407122); vascular endothelial growth factor antagonist (Borean, Denmark); bevacizumab (pINN) (Genentech, USA); angiogenic inhibitors (SUGEN, USA); XL 784 (Exelixis, USA); XL 647 (Exelixis, USA); MAb, integrin alpha5beta3, (second generation from Molecular Evolution, USA and USA); entazine hydrochloride (Lilly, USA); CEP55 (Cephalon, USA and Sfano-Sfynthelth, France); and BPI 21 (Cancer, Italy); (XOMA, USA); PI 88 (Progen, Australia); cilengitide (Merck KGaA, German; Technical University of Munich, Germany, Scripps Clinic and Research Foundation, USA); AVE 8062 (Ajinomoto, Japan); AS 1404 (Cancer Research Laboratory, New Zealand); SG 292, (Telios, USA); Endostatin (Boston Childrens Hospital, USA); ATN 161 (Attenuon, USA); 2-methoxyestradiol (Boston Childrens Hospital, USA); ZD 6474, (AstraZeneca, UK); ZD 6126, (Angiogene Pharmaceuticals, UK); PPI 2458, (Praecis, USA); AZD 9935, (AstraZeneca, UK); AZD 2171, (AstraZeneca, UK); vatalanib (pINN), (Novartis, Switzerland and Schering AG, Germany); tissue factor pathway inhibitors, (EntreMed, USA); pegaptanib (Pinn), (Gilead Sciences, USA); xanthorrizole, (Yonsei University, South Korea); VEGF-2 gene-based vaccine, (Scripps Clinic and Research Foundation, USA); SPV5.2, (Suratek, Canada); SDX 103, (University of California San Diego, USA); PX 478, (ProlX, USA); METASTATIN, (EntreMed, USA); troponin I, (Harvard University, USA); SU 6668, (SUGEN, USA); OXI 4503, (OXiGENE, USA); o-guanidines, (Dimensional Pharmaceuticals, USA); motuporamine C, (University of British Columbia, Canada); CDP 791, (Celltech Group, UK); atiprimod (pINN), (GlaxoSmithKline, UK); E 7820, (Eisai, Japan); CYC 381, (Harvard University, USA); AE 941, (Aeterna, Canada); vaccine, angiogenic, (EntreMed, USA); urokinase plasminogen activator inhibitor, (Dendreon, USA); oglufanide (pINN), (Melmotte, USA); HIF-lalpha inhibitors, (Xenova, UK); CEP 5214, (Cephalon, USA); BAY RES 2622, (Bayer, Germany); Angiocidin, (InKine, USA); A6, (Angstrom, USA); KR 31372, (Korea Research Institute of Chemical Technology, South Korea); GW 2286, (GlaxoSmithKline, UK); EHT 0101, (ExonHit, France); CP 868596, (Pfizer, USA); CP 564959, (OSI, USA); CP 547632, (Pfizer, USA); 786034, (GlaxoSmithKline, UK); KRN 633, (Kirin Brewery, Japan); drug delivery system, intraocular, 2-methoxyestradiol; anginex (Maastricht University, The Netherlands, and University of Minnesota, USA); ABT 510 (Abbott, USA); AAL 993 (Novartis, Switzerland); VEGI (ProteomTech, USA); tumor necrosis factor-alpha inhibitors; SU 11248 (Pfizer, USA and SUGEN USA); ABT 518, (Abbott, USA); YH16 (Yantai Rongchang, China); S-3APG (Boston Childrens Hospital, USA and EntreMed, USA); MAb, KDR (ImClone Systems, USA); MAb, alpha5 beta (Protein Design, USA); KDR kinase inhibitor (Celltech Group, UK, and Johnson & Johnson, USA); GFB 116 (University of South Florida, USA and Yale University, USA); CS 706 (Sankyo, Japan); combretastatin A4 prodrug (Arizona State University, USA); chondroitinase AC (IBEX, Canada); BAY RES 2690 (Bayer, Germany); AGM 1470 (Harvard University, USA, Takeda, Japan and TAP, USA); AG 13925 (Agouron, USA); Tetrathiomolybdate (University of Michigan, USA); GCS 100 (Wayne State University, USA) CV 247 (Ivy Medical, UK); CKD 732 (Chong Kun Dang, South Korea); irsogladin, (Nippon Shinyaku, Japan); RG 13577 (Aventis, France); WX 360 (Wilex, Germany); squalamine, (Genaera, USA); RPI 4610 (Sistema, USA); heparanase inhibitors (InSight, Israel); KL 3106 (Cologne, South Korea); Honokiol (Emory University, USA); ZK CDK (Schering AG, Germany); ZK Angio (Schering AG, Germany); ZK 229561 (Novartis, Switzerland, and Schering AG, Germany); XMP 300 (XOMA, USA); VGA 1102 (Taisho, Japan); VE-cadherin-2 antagonists (ImClone Systems, USA); Vasostatin (National Institutes of Health, USA); Flk-1 (ImClone Systems, USA); TZ 93 (Tsumura, Japan); TumStatin (Beth Israel Hospital, USA); truncated soluble FLT-1 (vascular endothelial growth factor receptor 1) (Merck & Co, USA); Tie-2 ligands (Regeneron, USA); and thrombospondin 1 inhibitor (Allegheny Health, Education and Research Foundation, USA).

[00243] Other examples of therapeutic agents that may be used in combination with compounds of the invention include agents (e.g., antibodies, antigen-binding regions, or soluble receptors) that specifically bind to and inhibit the activity of growth factors, such as antagonists of hepatocyte growth factor (HGF, also known as Scatter Factor) and antibodies or antigen-binding regions that specifically bind to its receptor, c-Met.

[00244] Another example of a therapeutic agent that may be used in combination with compounds of the invention is an autophagy inhibitor. Autophagy inhibitors include, but are not limited to, chloroquine, 3-methyladenine, hydroxychloroquine (Plaquenil™), bafilomycin A1, 5-amino-4-imidazole carboxamide riboside (AICAR), okadaic acid, autophagy-suppressing algal toxins that inhibit type 2A or type 1 protein phosphatases, cAMP analogs, and drugs that elevate cAMP levels, such as adenosine, LY204002, N6-mercaptopurine riboside, and vinblastine. Additionally, antisense or siRNA that inhibit the expression of proteins including, but not limited to, ATG5 (which are implicated in autophagy), may also be used. In some embodiments, one or more additional therapies include an autophagy inhibitor.

[00245] Another example of a therapeutic agent that may be used in combination with compounds of the invention is an antineoplastic agent. In some embodiments, one or more additional therapies include an antineoplastic agent. Non-limiting examples of antineoplastic agents include acemannan, aclarubicin, aldesleukin, alemtuzumab, alitretinoin, altretamine, amifostine, aminolevulinic acid, amrubicin, amsacrine, anagrelide, anastrozole, ancer, ancestim, arglabine, arsenic trioxide, BAM-002 (Novelos), bexarotene, bicalutamide, broxuridine, capecitabine, celmoleukin, cetrorelix, cladribine, clotrimazole, cytarabine ocphosphate, DA 3030 (Dong-A), daclizumab, denileukin diftitox, deslorelin, dexrazoxane, dilazep, docetaxel, docosanol, doxercalciferol, doxifluridine, doxorubicin, bromocriptine, carmustine, cytarabine, fluorouracil, diclofenac HIT, interferon alfa, daunorubicin, doxorubicin, tretinoin, edelfosine, edrecolomatine, eflornithine, emitefur, epirubicin, emitefur beta, etoposide phosphate, exemestane, exisulinde, fadrozole, filgrastim, finasteride, fludarabine phosphate, formestane, fotemustine, gallium nitrate, gemcitabine, gemtuzumab zogamicin, gimeracil/oteracil/tegafur combination, glycopin, goserelin, heptaplatin, human chorionic gonadotropin, human fetal alpha-fetoprotein, ibandronic acid, idarubicin, (imiquimod, interferon alfa, interferon alfa, natural, interferon alfa-2, interferon alfa-2a, interferon alfa-2b, interferon alfa-Nl, interferon alfa-n3, interferon alfacon-1, interferon alfa, natural, interferon beta, interferon beta-la, interferon beta-lb, interferon gamma, interferon gamma-la natural, interferon gamma-lb, interleukin-1 beta, iobenguane, irinotecan, irsogladine, lanreotide, LC 9018 (Yakult), leflunomide, lenograstim, lentinan sulfate, letrozole, leukocyte alpha interferon, leuprorelin, levamisole + fluorouracil, liarozole, lobaplatin, lonidamine, lovastatin, masoprocol, melarsoprol, metoclopramide, mifepristone, miltefosine, mirimostim, mismatched double-stranded RNA, mitoguazone, mitolactol, mitoxantrone, molgramostim, nafarelin, naloxone + pentazocine, nartograstim, nedaplatin, nilutamide, noscapine, novel erythropoiesis-stimulating protein, NSC 631570 octreotide, oprelvekin, osaterone, oxaliplatin, paclitaxel, pamidronate, pegaspargase, peginterferon alfa-2b, pentosan polysulfate sodium, pentostatin, picibanil, pirarubicin, rabbit polyclonal antithymocyte antibody, polyethylene glycol interferon alfa-2a, porfimer sodium, raloxifene, raltitrexed, rasburiebodiment, rhenium Re 186 etidronate, RII retinamide, rituximab, romurtide, samarium (153 Sm) lexidronam, sargramostim, sizofiran, sobuzoxane, sonermin, phentermine chloride strontium-89, suramin, tasonermin, tazarotene, tegafur, temoporfin, temozolomide, teniposide, tetrachlorodecaoxide, thalidomide, thymalfasin, thyrotropin alfa, topotecan, toremifene, tositumomab-iodine 131, trastuzumab, treosulfan, tretinoin, trilostane, trimetrexate, triptorelin, tumor necrosis factor alfa, natural, ubenimex, bladder cancer vaccine, Maruyama vaccine, melanoma lysate vaccine, valrubicin, verteporfin, vinorelbine, virulizine, zinostatin stimalamer or zoledronic acid; abarelix; AE 941 (Aeterna), ambamustine, antisense oligonucleotide, bcl-2 (Genta), APC 8015 (Dendreon), decitabine, dexaminoglutethimide, diaziquone, EL 532 (Elan), EM 800 (Endorecherche), eniluracil, etanidazole, fenretinide, filgrastim SD01 (Amgen), fulvestrant, gallocitabine, gastrin 17 immunogen, HLA-B7 gene therapy (Vical), granulocyte macrophage colony-stimulating factor, histamine dihydrochloride, ibritumomab tiuxetan, ilomastat, IM 862 (Cytran), interleukin-2, iproxifen, LDI 200 (Milkhaus), leridistim, lintuzumab, CA 125 MAb (Biomira), cancer MAb (Japan Pharmaceutical Development), HER-2 and Fc MAb (Medarex), idiotypic 105AD7 MAb (CRC Technology), idiotypic CEA MAb (Trilex), LYM-1-iodine 131 MAb (Clone Techni), polymorphic epithelial mucin-yttrium 90 MAb (Antisoma), marimastat, menogaril, mitumomab, motexafin gadolinium, MX 6 (Galderma), Nelarabine, Nolatrexed, Protein P 30, Pegvisomant, Pemetrexed, Porfiromycin, Prinomastat, RL 0903 (Shire), Rubitecan, Satraplatin, Sodium Phenylacetate, Sparphosic Acid, SRL 172 (SR Pharma), SU 5416 (SUGEN), TA 077 (Tanabe), Tetrathiomolybdate, taliblastin, thrombopoietin, ethyl tin etiopurpurin, tirapazamine, cancer vaccine (Biomira), melanoma vaccine (New York University), melanoma vaccine (Sloan Kettering Institute), oncolysed melanoma vaccine (New York Medical College), viral melanoma cell lysate vaccine (Royal Newcastle Hospital), or valspodar.

[00246] Additional examples of therapeutic agents that may be used in combination with compounds of the invention include ipilimumab (Yervoy®); tremelimumab; galiximab; nivolumab, also known as BMS-936558 (Opdivo®); pembrolizumab (Keytruda®); avelumab (Bavencio®); AMP224; BMS-936559; MPDL3280A, also known as RG7446; MEDI-570; AMG557; MGA271; IMP321; BMS-663513; PF-05082566; CDX-1127; anti-OX40 (Providence Health Services); huMAbOX40L; atacicept; CP-870893; lucatumumab; dacetuzumab; Muromonab-CD3; ipilumumab; MEDI4736 (Imfinzi®); MSB0010718C; AMP 224; adalimumab (Humira®); ado-trastuzumab emtansine (Kadcyla®); aflibercept (Eylea®); alemtuzumab (Campath®); basiliximab (Simulect®); belimumab (Benlysta®); basiliximab (Simulect®); belimumab (Benlysta®); brentuximab vedotin (Adcetris®); canakinumab (Ilaris®); Certolizumab pegal (Cimzia®); daclizumab (Zenapax®); daratumumab (Darzalex®); denosumab (Prolia®); eculizumab (Soliris®); efalizumab (Raptiva®); gemtuzumab ozogamicin (Mylotarg®); golimumab (Simponi®); ibritumomab tiuxetan (Zevalin®); infliximab (Remicade®); motavizumab (Numax®); natalizumab (Tysabri®); obinutuzumab (Gazyva®); ofatumumab (Arzerra®); omalizumab (Xolair®); palivizumab (Synagis®); pertuzumab (Perjeta®); pertuzumab (Perjeta®); ranibizumab (Lucentis®); raxibacumab (Abthrax®); tocilizumab (Actemra®); tositumomab; tositumomab-i-131; tositumomab and tositumomab-i-131 (Bexxar®); ustekinumab (Stelara®); AMG 102; AMG 386; AMG 479; AMG 655; AMG 706; AMG 745; and AMG 951.

[00247] The compounds described herein may be used in combination with the agents disclosed herein or other suitable agents, depending on the condition being treated. Therefore, in some embodiments, the one or more compounds of the disclosure will be co-administered with other therapies as described herein. When used in combination therapy, the compounds described herein may be administered with the second agent simultaneously or separately. Such combination administration may include simultaneous administration of the two agents in the same dosage form, simultaneous administration in separate dosage forms, and separate administration. That is, a compound described herein and any of the agents described herein may be formulated together in the same dosage form and administered simultaneously. Alternatively, a compound of the invention and any of the therapies described herein may be administered simultaneously, wherein both agents are present in separate formulations. In another alternative, a compound of the present disclosure may be administered and followed by any of the therapies described herein, or vice versa. In some embodiments of the separate administration protocol, a compound of the invention and any of the therapies described herein are administered a few minutes apart, or a few hours apart, or a few days apart.

[00248] In some embodiments of any of the methods described herein, the first therapy (e.g., a compound of the invention) and one or more additional therapies are administered simultaneously or sequentially, in any order. The first therapeutic agent can be administered immediately, up to 1 hour, up to 2 hours, up to 3 hours, up to 4 hours, up to 5 hours, up to 6 hours, up to 7 hours, up to 8 hours, up to 9 hours, up to 10 hours, up to 11 hours, up to 12 hours, up to 13 hours, 14 hours, up to 16 hours, up to 17 hours, up to 18 hours, up to 19 hours, up to 20 hours, up to 21 hours, up to 22 hours, up to 23 hours, up to 24 hours, or up to 1-7, 1-14, 1-21, or 1-30 days before or after one or more additional therapies.

[00249] The invention also features kits including (a) a pharmaceutical composition including an agent (e.g., a compound of the invention) described herein, and (b) a package insert with instructions for performing any of the methods described herein. In some embodiments, the kit includes (a) a pharmaceutical composition including an agent (e.g., a compound of the invention) described herein, (b) one or more additional therapies (e.g., non-drug treatment or therapeutic agent), and (c) a package insert with instructions for performing any of the methods described herein.

[00250] As one aspect of the present invention contemplates the treatment of disease or associated symptoms with a combination of pharmaceutically active compounds that can be administered separately, the invention further relates to the combination of separate pharmaceutical compositions in kit form. The kit may comprise two separate pharmaceutical compositions: a compound of the present invention and one or more additional therapies. The kit may comprise a container for holding the separate compositions, such as a divided bottle or a divided foil package. Additional examples of containers include syringes, boxes, and pouches. In some embodiments, the kit may comprise instructions for use of the separate components. The kit form is particularly advantageous when the separate components are preferably administered in different dosage forms (e.g., oral and parenteral), are administered at different dosage intervals, or when titration of the individual components of the combination is desired by the healthcare professional's prescription. LISTED EMBODIMENTS[1] A compound, or pharmaceutically acceptable salt thereof, having the structure of Formula I: Formula Iwhere the dotted lines represent zero, one, two, three, or four nonadjacent double bonds;A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 10-membered heteroarylene;B is absent, is -CH(R9)-, >C=CR9R9', or >CR9R9' where the carbon is bonded to the carbonyl carbon of -N(R11)C(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;G is optionally substituted C1-C4 alkylene, optionally substituted C1-C4 alkenylene, optionally substituted C1-C4 heteroalkylene, -C(O)O-CH(R6)- where C is attached to -C(R7R8)-, -C(O)NH-CH(R6)- where C is attached to -C(R7R8)-, optionally substituted C1-C4 heteroalkylene, or 3- to 8-membered heteroarylene; L is absent or is a ligand; W is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone, or an alkynyl sulfone; X 1 is optionally substituted C 1-C 2 alkylene, NR, O, or S(O) n ; X 2 is O or NH ; X 3 is N or CH ; n is 0, 1, or 2; R is hydrogen, cyano, optionally substituted C 1-C 4 alkyl, optionally substituted C 2-C 4 alkenyl, optionally substituted C 2-C 4 alkynyl, C(O) R', C(O) OR', C(O) N(R') 2 , S(O) R', S(O) 2R', or S(O) 2N(R') 2 ; each R' is independently H or optionally substituted C 1-C 4 alkyl; Y 1 is C, CH, or N ; Y 2 , Y 3 , Y 4 , and Y 7 are independently C or N ; Y 5 is CH, CH2, or N; Y6 is C(O), CH, CH2, or N; R1 is cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl, or R1 and R2 combine with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl; R2 is absent, is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 3- to 7-membered heteroaryl, optionally substituted 6-membered aryl, 5- or 6-membered optionally substituted; R3 is absent, orR2 and R3 combine with the atom to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or optionally substituted 3- to 14-membered heterocycloalkyl;R4 is absent, is hydrogen, halogen, cyano, or methyl optionally substituted with 1 to 3 halogens;R5 is hydrogen, C1-C4 alkyl optionally substituted with halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl, or cyclobutyl;R6 is hydrogen or methyl; R7 is hydrogen, halogen, or optionally substituted C1-C3 alkyl, or R6 and R7 combine with the carbon atoms to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7 and R8 combine with the carbon atom to which they are attached to form C=CR7'R8'; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;R7a and R8a are independently hydrogen, halo, C1-C3 alkyl or combine with the carbon to which they are attached to form a carbonyl; R7' is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R8' is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7' and R8' combine with the carbon atom to which they are attached to form optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R9 is H, F optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl, or R9 and L combine with the atoms to which they are attached to form form an optionally substituted 3- to 14-membered heterocycloalkyl; R9' is hydrogen or optionally substituted C1-C6 alkyl; or R9 and R9', combined with the atoms to which they are attached, form a 3- to 6-membered cycloalkyl or a 3- to 6-membered heterocycloalkyl; R10 is hydrogen, halo, hydroxy, C1-C3 alkoxy, or C1-C3 alkyl; R10a is hydrogen or halo; R11 is hydrogen or C1-C3 alkyl; and R21 is H or C1-C3 alkyl.[2] The compound, or pharmaceutically acceptable salt thereof, of paragraph [1], wherein G is optionally substituted C1-C4 heteroalkylene.[3] The compound, or pharmaceutically acceptable salt thereof, of paragraph [1] or [2], wherein the compound has the structure of Formula Ic: Formula Iwhere the dotted lines represent zero, one, two, three, or four nonadjacent double bonds;A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 6-membered heteroarylene;B is -CH(R9)- where the carbon is bonded to the carbonyl carbon of -N(R11)C(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;L is absent or is a ligand;W is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone, or a alkynyl sulfone;X2 is O or NH;X3 is N or CH;n is 0, 1 or 2; R is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, C(O)R', C(O)OR', C(O)N(R')2, S(O)R', S(O)2R', or S(O)2N(R')2;each R' is independently H or optionally substituted C1-C4 alkyl;Y1 is C, CH, or N;Y2, Y3, Y4, and Y7 are independently C or N;Y5 and Y6 are independently CH or N;R1 is cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, 6- to 10-membered aryl optionally substituted 5- to 10-membered heteroaryl; R2 is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl; R3 is absent, or R2 and R3 combine with the atom to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or optionally substituted 3- to 14-membered heterocycloalkyl; R4 is absent, is hydrogen, halogen, cyano, or methyl optionally substituted by 1 to 3 halogens; R5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl, or cyclobutyl; R6 is hydrogen or methyl; R7 is hydrogen, halogen, or optionally substituted C1-C3 alkyl, or R6 and R7 combine with the carbon atoms to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7 and R8 combine with the carbon atom to which they are attached to form C=CR7'R8'; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;R7' is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R8' is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7' and R8' combine with the carbon atom to which they are attached to form optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; R10 is hydrogen, hydroxy, C1-C3 alkoxy, or C1-C3 alkyl; and R11 is hydrogen or C1-C3 alkyl.[4] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [3], wherein X2 is NH.[5] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [4], wherein X3 is CH.[6] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [5], wherein R11 is hydrogen.[7] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [5], wherein R11 is C1-C3 alkyl.[8] The compound, or pharmaceutically acceptable salt thereof, of paragraph [7], wherein R11 is methyl.[9] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] to [6], wherein the compound has the structure of Formula Id: Formula Same where the dotted lines represent zero, one, two, three, or four nonadjacent double bonds;A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 6-membered heteroarylene;B is -CH(R9)- where the carbon is bonded to the carbonyl carbon of NHC(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;L is absent or is a ligand;W is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone, or an alkynyl sulfone;n is 0, 1, or 2;R is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2C4 alkenyl, optionally substituted C2-C4 alkynyl, C(O)R', C(O)OR', C(O)N(R')2, S(O)R', S(O)2R', or S(O)2N(R')2;each R' is independently H or optionally substituted C1-C4 alkyl;Y1 is C, CH, or N;Y2, Y3, Y4, and Y7 are independently C or N;Y5 and Y6 are independently CH or N;R1 is cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl substituted, optionally substituted 6- to 10-membered aryl or optionally substituted 5- to 10-membered heteroaryl; R2 is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl; R3 is absent, or R2 and R3 combine with the atom to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or optionally substituted 3- to 14-membered heterocycloalkyl; R4 is absent, is hydrogen, halogen, cyano, or methyl optionally substituted by 1 to 3 halogens; R5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl, or cyclobutyl; R6 is hydrogen or methyl; R7 is hydrogen, halogen, or optionally substituted C1-C3 alkyl, or R6 and R7 combine with the carbon atoms to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7 and R8 combine with the carbon atom to which they are attached to form C=CR7'R8'; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;R7' is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R8' is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7' and R8' combine with the carbon atom to which they are attached to form optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; and R10 is hydrogen, hydroxy, C1-C3 alkoxy, or C1-C3 alkyl.[10] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [9], wherein X1 is optionally substituted C1-C2 alkylene.[11] The compound, or pharmaceutically acceptable salt thereof, of paragraph [10], wherein X1 is methylene.[12] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [11], wherein R5 is hydrogen.[13] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [11], wherein R5 is C1-C4 alkyl optionally substituted by halogen.[14] The compound, or pharmaceutically acceptable salt thereof, of paragraph [13], wherein R5 is methyl.[15] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [14], wherein Y4 is C.[16] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [15], wherein R4 is hydrogen.[17] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [16], wherein Y5 is CH.[18] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [17], wherein Y6 is CH.[19] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [18], wherein Y1 is C.[20] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [19], wherein Y2 is C.[21] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] to [20], wherein Y3 is N.[22] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] to [21], wherein R3 is absent.[23] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] to [22], wherein Y7 is C.[24] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] to [6] or [9] to [23], wherein the compound has the structure of Formula Ie: Formula Ie where A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 6-membered heteroarylene; B is -CH(R9)- where the carbon is bonded to the carbonyl carbon of NHC(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene; L is absent or is a ligand; W is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone, or an alkynyl sulfone; R1 is cyano, optionally substituted C1-C6 alkyl, Optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl or optionally substituted 5- to 10-membered heteroaryl;R2 is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl; R3 is absent, or R2 and R3 combine with the atom to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or optionally substituted 3- to 14-membered heterocycloalkyl; R5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl or cyclobutyl; R6 is hydrogen or methyl; R7 is hydrogen, halogen, or optionally substituted C1-C3 alkyl, or R6 and R7 combine with the carbon atoms to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7 and R8 combine with the carbon atom to which they are attached to form C=CR7'R8'; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;R7' is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R8' is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7' and R8' combine with the carbon atom to which they are attached to form optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; and R10 is hydrogen, hydroxy, C1-C3 alkoxy, or C1-C3 alkyl.[25] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [3] through [24], wherein R6 is hydrogen.[26] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [25], wherein R2 is hydrogen, cyano, optionally substituted C1-C6 alkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 6-membered heterocycloalkyl.[27] The compound, or pharmaceutically acceptable salt thereof, of paragraph [26], wherein R2 is optionally substituted C1-C6 alkyl.[28] The compound, or pharmaceutically acceptable salt thereof, of paragraph [27], wherein R2 is ethyl.[29] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [28], wherein R7 is optionally substituted C1-C3 alkyl.[30] The compound, or pharmaceutically acceptable salt thereof, of paragraph 29 R7 is C1-C3 alkyl.[31] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through 30, wherein R8 is optionally substituted C1-C3 alkyl.[32] The compound, or pharmaceutically acceptable salt thereof, of paragraph [31], wherein R8 is C1-C3 alkyl.[33] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [32], wherein the compound has the structure of the formula If: Formula Ifem where A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 6-membered heteroarylene;B is -CH(R9)- where the carbon is bonded to the carbonyl carbon of NHC(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;L is absent or is a ligand;W is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone, or an alkynyl sulfone;R1 is cyano, optionally substituted C1-C6 alkyl, C1-C6 optionally substituted heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl; R2 is C1-C6 alkyl or 3- to 6-membered cycloalkyl; R7 is C1-C3 alkyl; R8 is C1-C3 alkyl; and R9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl.[34] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [33], wherein R1 is optionally substituted 6- to 10-membered aryl, optionally substituted 3- to 6-membered cycloalkenyl, or optionally substituted 5- to 10-membered heteroaryl.[35] The compound, or pharmaceutically acceptable salt thereof, of paragraph [34], wherein R1 is optionally substituted 6-membered aryl, optionally substituted 6-membered cycloalkenyl, or optionally substituted 6-membered heteroaryl.[36] The compound, or pharmaceutically acceptable salt thereof, of paragraph [35], wherein R1 is [37] The compound, or pharmaceutically acceptable salt thereof, of paragraph [36], wherein R1 is [38] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] to [37], wherein the compound has the structure of Formula Ig: Formula Igem where A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 6-membered heteroarylene; B is -CH(R9)- where the carbon is bonded to the carbonyl carbon of NHC(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene; L is absent or is a ligand; W is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone, or an alkynyl sulfone; R2 is C1-C6 alkyl, C1-C6 fluoroalkyl, or 3- to 6-membered cycloalkyl; R7 is C1-C3 alkyl; R8 is C1-C3 alkyl; and R9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl. Xe and Xf are independently N or CH; and R12 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl.[39] The compound, or pharmaceutically acceptable salt thereof, of paragraph [38], wherein Xe is N and Xf is CH.[40] The compound, or pharmaceutically acceptable salt thereof, of paragraph [38], wherein Xe is CH and Xf is N.[41] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [38] to [40], wherein R12 is optionally substituted C1-C6 heteroalkyl.[42] The compound, or pharmaceutically acceptable salt thereof, ÇH3of any one of paragraphs [38] to [41], wherein R12 is [43] The compound, or pharmaceutically acceptable salt thereof, of paragraph [1] or [2], wherein the compound has the structure of Formula VI: wherein the dotted lines represent zero, one, two, three, or four nonadjacent double bonds; A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 10-membered heteroarylene; B is absent, is -CH(R9)-, >C=CR9R9', or >CR9R9' where the carbon is bonded to the carbonyl carbon of -N(R11)C(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene; G is optionally substituted C1-C4 alkylene, Optionally substituted C1-C4 alkenylene, optionally substituted C1-C4 heteroalkylene, -C(O)O-CH(R6)- where C is attached to -C(R7R8)-, -C(O)NH-CH(R6)- where C is attached to -C(R7R8)-, optionally substituted C1-C4 heteroalkylene, or 3- to 8-membered heteroarylene;L is absent or is a ligand;W is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone, a haloacetal, or an alkynyl sulfone;X1 is optionally substituted C1-C2 alkylene, NR, O, or S(O)n;X2 is O or NH;X3 is N or CH;n is 0, 1, or 2;R is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, C2-C4 optionally substituted alkynyl, C(O)R', C(O)OR', C(O)N(R')2, S(O)R', S(O)2R', or S(O)2N(R')2; each R' is independently H or optionally substituted C1-C4 alkyl; Y 1 is C, CH, or N; Y 2, Y3, Y4, and Y7 are independently C or N; Y 5 is CH, CH2, or N; Y6 is C(O), CH, CH2, or N; R2 is absent, is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl; R3 is absent, or R2 and R3 combine with the atom to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or optionally substituted 3- to 14-membered heterocycloalkyl; R4 is absent, is hydrogen, halogen, cyano, or methyl optionally substituted by 1 to 3 halogens; R5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl, or cyclobutyl; R6 is hydrogen or methyl; R7 is hydrogen, halogen, or optionally substituted C1-C3 alkyl, or R6 and R7 combine with the carbon atoms to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7 and R8 combine with the carbon atom to which they are attached to form C=CR7'R8'; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;R7a and R8a are independently hydrogen, halo, C1-C3 alkyl, or combine with the carbon to which they are attached to form a carbonyl;R7' is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R8' is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7' and R8' combine with the carbon atom to which they are attached to form optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R9 is H, F optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; or R9 and L combine with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl; R9' is hydrogen or optionally substituted C1-C6 alkyl; or R9 and R9', combined with the atoms to which they are attached, form a 3- to 6-membered cycloalkyl or a 3- to 6-membered heterocycloalkyl; R10 is hydrogen, halo, hydroxy, C1-C3 alkoxy, or C1-C3 alkyl; R10a is hydrogen or halo; R11 is hydrogen or C1-C3 alkyl; R21 is hydrogen or C1-C3 alkyl (e.g., methyl); and Xe and Xf are, independently, N or CH.[44] The compound, or pharmaceutically acceptable salt thereof, of paragraph [43], wherein the compound has the structure of Formula VIa: Formula VIawhere A is, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 6-membered heteroarylene;B is -CH(R9)- where the carbon is bonded to the carbonyl carbon of NHC(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;L is absent or is a ligand;W is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone, or an alkynyl sulfone;X1 is optionally substituted C1-C2 alkylene, NR, O, or S(O)n;X2 is O or NH;n is 0, 1, or 2;R is hydrogen, cyano, C1-C4 optionally substituted alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, C(O)R', C(O)OR', C(O)N(R')2, S(O)R', S(O)2R', or S(O)2N(R')2; each R' is independently H or optionally substituted C1-C4 alkyl; R2 is C1-C6 alkyl, C1-C6 fluoroalkyl, or 3- to 6-membered cycloalkyl; R7 is C1-C3 alkyl; R8 is C1-C3 alkyl; and R9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered heterocycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; Xe and Xf are independently N or CH; R11 is hydrogen or C1-C3 alkyl; and R21 is hydrogen or C1-C3 alkyl.[45] The compound, or pharmaceutically acceptable salt thereof, of paragraph [43] or [44], wherein the compound has the structure of Formula VIb: Formula VIwhere A is optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 6-membered heteroarylene;B is -CH(R9)- where the carbon is bonded to the carbonyl carbon of NHC(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;R9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;L is absent or is a ligand; andW is a crosslinking group comprising a vinyl ketone, a vinyl sulfone, an ynone, or an alkynyl sulfone.[46] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] to [45], wherein A is optionally substituted 6-membered arylene.[47] The compound, or pharmaceutically acceptable salt thereof, of paragraph [46], wherein A has the structure: wherein R13 is hydrogen, halo, hydroxy, amino, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; and R13a is hydrogen or halo.[48] The compound, or pharmaceutically acceptable salt thereof, of paragraph [47], wherein R13 and R13a are each hydrogen.[49] The compound, or pharmaceutically acceptable salt thereof, of paragraph [47], wherein R13 is hydroxy, methyl, fluoro, or difluoromethyl.[50] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [45], wherein A is optionally substituted 5- to 6-membered heteroarylene.[51] The compound, or pharmaceutically acceptable salt thereof, of paragraph [50], wherein A is: [52] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] to [45], wherein A is optionally substituted C1-C4 heteroalkylene.[53] The compound, or pharmaceutically acceptable salt thereof, of paragraph [52], wherein A is: [54] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] to [45], wherein A is optionally substituted 3- to 6-membered heterocycloalkylene.[55] The compound, or pharmaceutically acceptable salt thereof, of paragraph [54], wherein A is: [56] The compound, or pharmaceutically acceptable salt thereof, of paragraph [55], wherein A is [57] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] to [56], wherein B is -CHR9-.[58] The compound, or pharmaceutically acceptable salt thereof, of paragraph [57], wherein R9 is F, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl.[59] The compound, or pharmaceutically acceptable salt thereof, of paragraph [58], wherein R9 is: [60] The compound, or pharmaceutically acceptable salt, of paragraph [59], wherein R9 is: [61] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] to [56], wherein B is optionally substituted 6-membered arylene.[62] The compound, or pharmaceutically acceptable salt thereof, of paragraph [61], wherein B is 6-membered arylene.[63] The compound, or pharmaceutically acceptable salt thereof, of paragraph [61], wherein B is: [64] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [63], wherein R7 is methyl.[65] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [64], wherein R8 is methyl.[66] The compound, or pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [65], wherein the linker is the structure of Formula II:A1-(B1)f-(C1)g-(B2)h-(D1)-(B3)i-(C2)j-(B4)k-A2 Formula IIwhere A1 is a bond between the linker and B; A2 is a bond between W and the linker; B1, B2, B3, and B4 are each independently selected from optionally substituted C1-C2 alkylene, optionally substituted C1-C3 heteroalkylene, O, S, and NRN; RN is hydrogen, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted C1-C7 heteroalkyl; C1 and C2 are each independently selected from carbonyl, thiocarbonyl, sulfonyl, or phosphoryl; f, g, h, i, j, and k are each independently 0 or 1; and D1 is optionally substituted C1-C10 alkylene, optionally substituted C2-C10 alkenylene, optionally substituted C2-C10 alkynylene, optionally substituted 3- to 14-membered heterocycloalkylene, optionally substituted 5- to 10-membered heteroarylene, optionally substituted 3- to 8-membered cycloalkylene, optionally substituted 6- to 10-membered arylene, optionally substituted C2-C10 polyethylene glycolene, or optionally substituted C1-C10 heteroalkylene, or a chemical bond linking A1-(B1)f-(C1)g-(B2)h- to -(B3)i-(C2)j-(B4)k-A2.[67] The compound, or a pharmaceutically acceptable salt thereof, of any one of paragraphs [1] to [66], wherein the linker is acyclic.[68] The compound, or a pharmaceutically acceptable salt thereof, of paragraph [67], wherein the ligand has the structure of Formula IIa: Formula IIa in which Xa is absent or is N; R14 is absent, is hydrogen or optionally substituted C1-C6 alkyl; and L2 is absent, is -SO2-, optionally substituted C1-C4 alkylene or optionally substituted C1-C4 heteroalkylene, in which at least one of Xa, R14 or L2 is present.[69] The compound, or a pharmaceutically acceptable salt thereof, of paragraph [68], wherein the ligand has the structure: [70] The compound, or a pharmaceutically acceptable salt thereof, of any one of paragraphs [1] to [66], wherein the linker is or comprises a cyclic moiety.[71] The compound, or a pharmaceutically acceptable salt thereof, of paragraph [70], wherein the linker has the structure of Formula IIb: Formula IIb wherein o is 0 or 1; R15 is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted 3- to 8-membered cycloalkylene, or optionally substituted 3- to 8-membered heterocycloalkylene; X4 is absent, is optionally substituted C1-C4 alkylene, O, NCH3, or optionally substituted C1-C4 heteroalkylene; Cy is optionally substituted 3- to 8-membered cycloalkylene, optionally substituted 3- to 8-membered heterocycloalkylene, optionally substituted 6- to 10-membered arylene, or optionally substituted 5- to 10-membered heteroarylene; L3 is absent, is -SO2-, optionally substituted C1-C4 alkylene, or optionally substituted C1-C4 heteroalkylene.[72] The compound, or a pharmaceutically acceptable salt thereof, of paragraph [71], wherein the linker has the structure: Formula IIce wherein R15 is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted 3- to 8-membered cycloalkylene, or optionally substituted 3- to 8-membered heterocycloalkylene; and R15a, R15b, R15c, R15d, R15e, R15f, and R15g are independently hydrogen, halo, hydroxy, cyano, amino, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, or R15b and R15d combine with the carbons to which they are attached to form an optionally substituted 3- to 8-membered cycloalkylene or optionally substituted 3- to 8-membered heterocycloalkylene.[73] The compound, or a pharmaceutically acceptable salt thereof, of paragraph [72], wherein the linker has the structure: [74] The compound, or a pharmaceutically acceptable salt thereof, of paragraph [71], wherein the linker has the structure: [75] The compound, or a pharmaceutically acceptable salt thereof, of any one of paragraphs [1] to [74], wherein W is a crosslinking group comprising a vinyl ketone.[76] . The compound, or a pharmaceutically acceptable salt thereof, of paragraph [75], wherein W has the structure of Formula IIIa: Formula IIIawherein R16a, R16b, and R16c are independently hydrogen, -CN, halogen, or -C1-C3 alkyl optionally substituted by one or more substituents independently selected from -OH, -O-C1-C3 alkyl, -NH2, -NH(C1-C3 alkyl), -N(C1-C3 alkyl)2, or a 4- to 7-membered saturated heterocycloalkyl.[77] The compound, or a pharmaceutically acceptable salt thereof, of paragraph [76], wherein W is: [78] The compound, or a pharmaceutically acceptable salt thereof, of any one of paragraphs [1] to [74], wherein W is a crosslinking group comprising an inone.[79] The compound, or a pharmaceutically acceptable salt thereof, of paragraph [78], wherein W has the structure of Formula IIIb: Formula IIIb wherein R17 is hydrogen, -C1-C3 alkyl optionally substituted by one or more substituents independently selected from -OH, -O-C1-C3 alkyl, -NH2, -NH(C1-C3 alkyl), -N(C1-C3 alkyl)2, or a saturated 4- to 7-membered cycloalkyl, or a saturated 4- to 7-membered heterocycloalkyl.[80] The compound, or a pharmaceutically acceptable salt thereof, of paragraph [79], wherein W is: [81] The compound, or a pharmaceutically acceptable salt thereof, of any one of paragraphs [1] to [74], wherein W is a crosslinking group comprising a vinyl sulfone.[82] The compound, or a pharmaceutically acceptable salt thereof, of paragraph [81], wherein W has the structure of Formula IIIc: Formula IIIc wherein R18a, R18b, and R18c are independently hydrogen, -CN or -C1-C3 alkyl optionally substituted by one or more substituents independently selected from -OH, -O-C1-C3 alkyl, -NH2, -NH(C1-C3 alkyl), -N(C1-C3 alkyl)2, or a 4- to 7-membered saturated heterocycloalkyl.[83] The compound, or a pharmaceutically acceptable salt thereof, of paragraph [82], wherein W is: [84] The compound, or a pharmaceutically acceptable salt thereof, of any one of paragraphs [1] to [74], wherein W is a crosslinking group comprising an alkynyl sulfone.[85] The compound, or a pharmaceutically acceptable salt thereof, of paragraph [84], wherein W has the structure of Formula IIId: Formula III wherein R19 is hydrogen, -C1-C3 alkyl optionally substituted by one or more substituents independently selected from OH, -O-C1-C3 alkyl, -NH2, -NH(C1-C3 alkyl), -N(C1-C3 alkyl)2, or a 4- to 7-membered saturated heterocycloalkyl, or a 4- to 7-membered saturated heterocycloalkyl.[86] The compound, or a pharmaceutically acceptable salt thereof, of paragraph [85], wherein W is: [87] The compound, or a pharmaceutically acceptable salt thereof, of any one of paragraphs [1] to [74], wherein W has the structure of Formula IIIe: Formula IIIe wherein Xe is a halogen; and R20 is hydrogen, -C1-C3 alkyl optionally substituted by one or more substituents independently selected from -OH, -O-C1-C3 alkyl, -NH2, -NH(C1-C3 alkyl), -N(C1-C3 alkyl)2, or a saturated 4- to 7-membered heterocycloalkyl.[88] A compound, or a pharmaceutically acceptable salt thereof, selected from Table 1 or Table 2.[89] A pharmaceutical composition comprising a compound, or a pharmaceutically acceptable salt thereof, of any one of paragraphs [1] to [88], and a pharmaceutically acceptable excipient.[90] A conjugate, or salt thereof, comprising the structure of Formula IV:ML-PFormula IVwherein L is a linker; P is a monovalent organic moiety; and M has the structure of Formula V: Formula The dotted lines represent zero, one, two, three, or four nonadjacent double bonds;A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 6-membered heteroarylene;B is absent, is -CH(R9)-, >C=CR9R9', or >CR9R9' where the carbon is bonded to the carbonyl carbon of -N(R11)C(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;G is optionally substituted C1-C4 alkylene, Optionally substituted C1-C4 alkenylene, optionally substituted C1-C4 heteroalkylene, -C(O)O-CH(R6)- where C is attached to -C(R7R8)-, -C(O)NH-CH(R6)- where C is attached to -C(R7R8)-, optionally substituted C1-C4 heteroalkylene, or 3- to 8-membered heteroarylene;X1 is optionally substituted C1-C2 alkylene, NR, O, or S(O)n;X2 is O or NH;X3 is N or CH;n is 0, 1, or 2;R is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2C4 alkenyl, optionally substituted C2-C4 alkynyl, C(O)R', C(O)OR', C(O)N(R')2, S(O)R', S(O)2R', or S(O)2N(R')2;each R' is independently H or optionally substituted C1-C4 alkyl;Y1 is C, CH, or N;Y2, Y3, Y4, and Y7 are independently C or N;Y5 is CH, CH2, or N;Y6 is C(O), CH, CH2, or N;R1 is cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl, or R1 and R2 combine with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl;R2 is absent, is hydrogen, optionally substituted C1-C6 alkyl, Optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl; R3 is absent, or R2 and R3 combine with the atom to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or optionally substituted 3- to 14-membered heterocycloalkyl; R4 is absent, is hydrogen, halogen, cyano, or methyl optionally substituted by 1 to 3 halogens; R5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl, or cyclobutyl; R6 is hydrogen or methyl; R7 is hydrogen, halogen, or optionally substituted C1-C3 alkyl, or R6 and R7 combine with the carbon atoms to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7 and R8 combine with the carbon atom to which they are attached to form C=CR7'R8'; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;R7a and R8a are independently hydrogen, halo, C1-C3 alkyl, or combine with the carbon to which they are attached to form a carbonyl;R7' is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R8' is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7' and R8' combine with the carbon atom to which they are attached to form optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R9 is H, F optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl, or R9 and L combine with the atoms to which they are attached to form form an optionally substituted 3- to 14-membered heterocycloalkyl; R9' is hydrogen or optionally substituted C1-C6 alkyl; or R9 and R9', combined with the atoms to which they are fixedly attached, form a 3- to 6-membered cycloalkyl or a 3- to 6-membered heterocycloalkyl; R10 is hydrogen, halo, hydroxy, C1-C3 alkoxy, or C1-C3 alkyl; R10a is hydrogen or halo; R11 is hydrogen or C1-C3 alkyl; and R21 is H or C1-C3 alkyl.[91] . The conjugate of paragraph 90, or salt thereof, wherein M has the structure of formula Vd: Formula Vwhere A is, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 6-membered heteroarylene;B is -CH(R9)- where the carbon is bonded to the carbonyl carbon of NHC(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;X1 is optionally substituted C1-C2 alkylene, NR, O, or S(O)n;X2 is O or NH;n is 0, 1, or 2;R is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, C(O)R', C(O)OR', C(O)N(R')2, S(O)R', S(O)2R', or S(O)2N(R')2; each R' is independently H or optionally substituted C1-C4 alkyl; R2 is C1-C6 alkyl, C1-C6 fluoroalkyl, or 3- to 6-membered cycloalkyl; R7 is C1-C3 alkyl; R8 is C1-C3 alkyl; and R9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; Xe and Xf are independently N or CH; R11 is hydrogen or C1-C3 alkyl; and R21 is hydrogen or C1-C3 alkyl.[92] The conjugate of paragraph [91], or salt thereof, wherein M has the structure of Formula Ve: Formula Ve where A is, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 6-membered heteroarylene; B is -CH(R9)- where the carbon is bonded to the carbonyl carbon of NHC(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene; and R9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl.[93] The conjugate, or salt thereof, of any one of paragraphs [90] to [92], wherein the ligand has the structure of Formula II:A1 -(B1)f-(C1)g-(B2)h-(D1 )-(B3)i-(C2)j-(B4)k-A2Formula IIwhere A1 is a bond between the ligand and B; A2is a bond between W and the ligand; B1, B2, B3, and B4 each, independently, are selected from optionally substituted C1-C2 alkylene, optionally substituted C1-C3 heteroalkylene, O, S, and NRN; RN is hydrogen, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 aquinyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted C1-C7 heteroalkyl; C1 and C2 are each independently selected from carbonyl, thiocarbonyl, sulfonyl, or phosphoryl; f, g, h, i, j, and k are each independently 0 or 1; and D1 is optionally substituted C1-C10 alkylene, optionally substituted C2-C10 alkenylene, optionally substituted C2C10 alkynylene, optionally substituted 3- to 14-membered heterocycloalkylene, optionally substituted 5- to 10-membered heteroarylene, optionally substituted 3- to 8-membered cycloalkylene, optionally substituted 6- to 10-membered arylene, optionally substituted C2-C10 polyethylene glycolene, or optionally substituted C1-C10 heteroalkylene, or a chemical bond linking A1-(B1)f-(C1)g-(B2)h- to -(B3)i-(C2)j-(B4)k-A2.[94] The conjugate, or salt thereof, of any one of paragraphs [90] to [93], wherein the monovalent organic moiety is a protein.[95] The conjugate, or salt thereof, of paragraph [94], wherein the protein is a Ras protein.[96] The conjugate, or salt thereof, of paragraph [95], wherein the Ras protein is K-Ras G12C, K-Ras G13C, H-Ras G12C, H-Ras G13C, N-Ras G12C, or N-Ras G13C.[97] The conjugate, or salt thereof, of any one of paragraphs [93] through [96], wherein the linker is attached to the monovalent organic moiety through a bond to a sulfhydryl group of an amino acid residue of the monovalent organic moiety.[98] A method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound, or a pharmaceutically acceptable salt thereof, of any one of paragraphs [1] through [88] or a pharmaceutical composition of paragraph [89].[99] The method of paragraph [98], wherein the cancer is pancreatic cancer, colorectal cancer, non-small cell lung cancer, or endometrial cancer.[100] The method of paragraph [98] or [99], wherein the cancer comprises a Ras mutation.[101] The method of paragraph [100], wherein the Ras mutation is KRas G12C, K-Ras G13C, H-Ras G12C, H-Ras G13C, N-Ras G12C, or N-Ras G13C.[102] A method of treating a Ras protein-related disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound, or a pharmaceutically acceptable salt thereof, of any one of paragraphs [1] to [88] or a pharmaceutical composition of paragraph [89].[103] A method of inhibiting a Ras protein in a cell, the method comprising contacting the cell with an effective amount of a compound, or a pharmaceutically acceptable salt thereof, of any one of paragraphs [1] to [88] or a pharmaceutical composition of paragraph [89].[104] The method of paragraph [102] or [103], wherein the Ras protein is K-Ras G12C, K-Ras G13C, H-Ras G12C, H-Ras G13C, N-Ras G12C, or N-Ras G13C.[105] The method of paragraph [103] or [104], wherein the cell is a cancer cell.[106] The method of paragraph [105], wherein the cancer cell is a pancreatic cancer cell, a colorectal cancer cell, a non-small cell lung cancer cell, or an endometrial cancer cell.[107] The method or use of any one of paragraphs [98] to [106], wherein the method or use further comprises administering an additional anti-cancer therapy.[108] The method of paragraph [107], wherein the additional anti-cancer therapy is an EGFR inhibitor, a second Ras inhibitor, a SHP2 inhibitor, a SOS1 inhibitor, a Raf inhibitor, a MEK inhibitor, an ERK inhibitor, a PI3K inhibitor, a PTEN inhibitor, an AKT inhibitor, an mTORC1 inhibitor, a BRAF inhibitor, a PD-L1 inhibitor, a PD-1 inhibitor, a CDK4/6 inhibitor, a HER2 inhibitor, or a combination thereof.[109] The method of paragraph [107] or [108], wherein the additional anti-cancer therapy is a SHP2 inhibitor.

EXAMPLES

[00251] The disclosure is further illustrated by the following examples and summary examples, which are not to be construed as limiting this disclosure in scope or spirit to the specific procedures described herein. It should be understood that the examples are provided to illustrate particular embodiments and that no limitation of the scope of this disclosure is intended thereby. It should also be understood that various other embodiments, modifications and equivalents thereof may be available, which may suggest the same to those skilled in the art, without departing from the spirit of the present disclosure or the scope of the appended claims.

CHEMICAL SYNTHESIS

[00252] The definitions used in the following examples and elsewhere in this document are: CH2Cl2, DCM Methylene chloride, Dichloromethane CH3CN, MeCN Acetonitrile CuI Copper(I) iodide DIPEA Diisopropylethyl amine DMF N,N-Dimethylformamide EtOAc Ethyl acetate H2O Water HCI Hydrochloric acid K3PO4 Potassium phosphate (tribasic) MeOH Methanol Na2SO4 Sodium sulfate NMP N-methyl pyrrolidone Pd(dppf)Cl2 [1,1'-Bis(diphenylphosphino)ferrocene]dichloropalladium(II)

INSTRUMENTATION

[00253] Mass spectrometry data collection occurred with a Shimadzu LCMS-2020, an Agilent 1260LC-6120/6125MSD, a Shimadzu LCMS-2010EV, or a Waters Acquity UPLC, with a QDa detector or SQ 2 detector. Samples were injected in their liquid phase onto a C-18 reversed phase. Compounds were eluted from the column using an acetonitrile gradient and fed to the mass analyzer. Initial data analysis occurred with Agilent ChemStation, Shimadzu LabSolutions, or Waters MassLynx. NMR data were collected with a Bruker AVANCE III HD 400MHz, a Bruker Ascend 500MHz or a Varian 400MHz instrument, and raw data were analyzed with TopSpin or Mestrelab Mnova.SYNTHESIS OF INTERMEDIATESINTERMEDIATE 1. SYNTHESIS OF 3-(5-BROMO-1-ETHYL-2-[2-[(1S)-1-METHOXYETHYL]PYRIDIN-3-YL]INDOL-3-YL)-2,2-DIMETHYLPROPAN-1-OL

[00254] Step 1. To a mixture of 3-(tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropanoyl chloride (65 g, 137 mmol, crude) in DCM (120 mL) at 0 °C under a N2 atmosphere was added 1 M SnCl4 in DCM (137 mL, 137 mmol) slowly. The mixture was stirred at 0 °C for 30 min, then a solution of 5-bromo-1H-indole (26.8 g, 137 mmol) in DCM (40 mL) was added dropwise. The mixture was stirred at 0 °C for 45 min, then diluted with EtOAc (300 mL), washed with brine (100 mL × 4), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 1-(5-bromo-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropan-1-one (55 g, 75% yield). LCMS (ESI): m/z [M+Na] calcd for C29H32BrNO2SiNa 556.1; found 556.3.

[00255] Step 2. To a mixture of 1-(5-bromo-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropan-1-one (50 g, 93.6 mmol) in THF (100 mL) at 0 °C under a N2 atmosphere was added LiBH4 (6.1 g, 281 mmol). The mixture was heated to 60 °C and stirred for 20 h, then MeOH (10 mL) and EtOAc (100 mL) were added and the mixture was washed with brine (50 mL), dried over Na2SO4, filtered, and the filtrate concentrated under reduced pressure. The residue was diluted with DCM (50 mL), cooled to 10 °C, and diludine (9.5 g, 37.4 mmol) and TsOH.H2O (890 mg, 4.7 mmol) added. The mixture was stirred at 10 °C for 2 h, filtered, the filtrate concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give 1-(5-bromo-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropan-1-one (41 g, 84% yield). LCMS (ESI): m/z [M+H] calcd for C29H34BrNOSi 519.2; found 520.1; 1H NMR (400 MHz, CDCl3) δ 7.96 (s, 1H), 7.75 - 7.68 (m, 5H), 7.46 - 7.35 (m, 6H), 7.23 - 7.19 (m, 2H), 6.87 (d, J = 2.1 Hz, 1H), 3.40 (s, 2H), 2.72 (s, 2H), 1.14 (s, 9H), 0.89 (s, 6H).

[00256] Step 3. To a mixture of 1-(5-bromo-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropan-1-one (1.5 g, 2.9 mmol) and I2 (731 mg, 2.9 mmol) in THF (15 mL) at rt was added AgOTf (888 mg, 3.5 mmol). The mixture was stirred at rt for 2 h, then diluted with EtOAc (200 mL) and washed with saturated Na2S2O3 (100 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-iodo-1H-indole (900 mg, 72% yield) as a solid. 1H NMR (400 MHz, DMSO-d6) δ 11.70 (s, 1H), 7.68 (d, J = 1.3 Hz, 1H), 7.64 - 7.62 (m, 4H), 7.46 - 7.43 (m, 6H), 7.24 - 7.22 (d, 1H), 7.14 - 7.12 (dd, J = 8.6, 1.6 Hz, 1H), 3.48 (s, 2H), 2.63 (s, 2H), 1.08 (s, 9H), 0.88 (s, 6H).

[00257] Step 4. To a stirred mixture of HCOOH (66.3 g, 1.44 mol) in TEA (728 g, 7.2 mol) at 0 °C under an Ar atmosphere was added (4S,5S)-2-chloro-2-methyl-1-(4-methylbenzenesulfonyl)-4,5-diphenyl-1,3-diaza-2-ruthenacyclopentanecimene (3.9 g, 6.0 mmol) in portions. The mixture was heated to 40 °C and stirred for 15 min, then cooled to rt and 1-(3-bromopyridin-2-yl)ethanone (120 g, 600 mmol) added in portions. The mixture was heated to 40 °C and stirred for an additional 2 h, then the solvent was concentrated under reduced pressure. Brine (2 L) was added to the residue, the mixture was extracted with EtOAc (4 × 700 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give (1S)-1-(3-bromopyridin-2-yl)ethanol (100 g, 74% yield) as an oil. LCMS (ESI): m/z [M+H] calculated for C7H8BrNO 201.1; found 201.9.

[00258] Step 5. To a stirred mixture of (1S)-1-(3-bromopyridin-2-yl)ethanol (100 g, 495 mmol) in DMF (1 L) at 0 °C was added NaH, 60% dispersion in oil (14.25 g, 594 mmol) in portions. The mixture was stirred at 0 °C for 1 h. MeI (140.5 g, 990 mmol) was added dropwise at 0 °C and the mixture was allowed to warm to rt and stirred for 2 h. The mixture was cooled to 0 °C and saturated NH 4 Cl (5 L) was added. The mixture was extracted with EtOAc (3 x 1.5 L), dried over anhydrous Na 2 SO 4 , and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give 3-bromo-2-[(1S)-1-methoxyethyl]pyridine (90 g, 75% yield) as an oil. LCMS (ESI): m/z [M+H] calcd for C8H10BrNO 215.0; found 215.9.

[00259] Step 6. To a stirred mixture of 3-bromo-2-[(1 S)-1-methoxyethyl]pyridine (90 g, 417 mmol) and Pd(dppf)Cl2 (30.5 g, 41.7 mmol) in toluene (900 mL) at rt under an Ar atmosphere was added bis(pinacolato)diboron (127 g, 500 mmol) and KOAc (81.8 g, 833 mmol) in portions. The mixture was heated to 100 °C and stirred for 3 h. The filtrate was concentrated under reduced pressure and the residue was purified by Al2O3 column chromatography to obtain 2-[(1S)-1-methoxyethyl]-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (100 g, 63% yield) as a semisolid. LCMS (ESI): m/z [M+H] calcd for C14H22BNO3 263.2; found 264.1.

[00260] Step 7. To a stirred mixture of 5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-2-iodo-1H-indole (140 g, 217 mmol) and 2-[(1S)-1- methoxyethyl]-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (100 g, 380 mmol) in 1,4-dioxane (1.4 L) at rt under an Ar atmosphere were added K2CO3 (74.8 g, 541 mmol), Pd(dppf)Cl2 (15.9 g, 21.7 mmol) and H2O (280 mL) in portions. The mixture was heated to 85 °C and stirred for 4 h, then cooled, H2O (5 L) added, and the mixture extracted with EtOAc (3 × 2 L). The combined organic layers were washed with brine (2 × 1 L), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give 5-bromo-3-[3-tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-1H-indole (71 g, 45% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C37H43BrN2O2Si 654.2; found 655.1.

[00261] Step 8. To a stirred mixture of 5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-2-[2-[1S)-1-methoxyethyl]pyridin-3-yl]-1H]-indole (71 g, 108 mmol) in DMF (0.8 L) at 0 °C under a N2 atmosphere was added Cs2CO3 (70.6 g, 217 mmol) and EtI (33.8 g, 217 mmol) in portions. The mixture was warmed to rt and stirred for 16 h, then H2O (4 L) was added and the mixture was extracted with EtOAc (3 × 1.5 L). The combined organic layers were washed with brine (2 × 1 L), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indole (66 g, 80% yield) as an oil. LCMS (ESI): m/z [M+H] calcd for C39H47BrN2O2Si 682.3; found 683.3.

[00262] Step 9: To a stirred mixture of TBAF (172.6 g, 660 mmol) in THF (660 mL) at rt under a N2 atmosphere was added 5-bromo-3-[3-[tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indole (66 g, 97 mmol) in portions. The mixture was heated to 50 °C and stirred for 16 h, cooled, diluted with H2O (5 L), and extracted with EtOAc (3 × 1.5 L). The combined organic layers were washed with brine (2 × 1 L), dried over anhydrous Na2SO4, and filtered. After filtration, the filtrate was concentrated under reduced pressure. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to obtain 3-(5-bromo-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-3-yl)-2,2-dimethylpropan-1-ol (30 g, 62% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C23H29BrN2O2 444.1; found 445.1. INTERMEDIATE 1. ALTERNATIVE SYNTHESIS VIA THE FISHER INDOL ROUTE.

[00263] Step 1. To a mixture of i-PrMgCl (2M in THF, 0.5 L) at -10 °C under a N 2 atmosphere was added n-BuLi , 2.5 M in hexane (333 mL, 833 mmol) dropwise over 15 min. The mixture was stirred for 30 min at -10 °C and then 3-bromo-2-[(1S)-1-methoxyethyl]pyridine (180 g, 833 mmol) in THF (0.5 L) was added dropwise over 30 min at -10 °C. The resulting mixture was warmed to -5 °C and stirred for 1 h, then 3,3-dimethyloxane-2,6-dione (118 g, 833 mmol) in THF (1.2 L) was added dropwise over 30 min at -5 °C. The mixture was warmed to 0 °C and stirred for 1.5 h, then quenched with the addition of pre-chilled 4 M HCl in 1,4-dioxane (0.6 L) at 0 °C to adjust the pH ~5. The mixture was diluted with ice-cold water (3 L) and extracted with EtOAc (3 × 2.5 L). The combined organic layers were dried over anhydrous Na 2 SO 4 , filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give 5-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-2,2-dimethyl-5-oxopentanoic acid (87 g, 34% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C 15 H 21 NO 4 279.2; found 280.1.

[00264] Step 2. To a mixture of 5-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-2,2-dimethyl-5-oxopentanoic acid (78 g, 279 mmol) in EtOH (0.78 L) at rt under a N2 atmosphere was added (4-bromophenyl)hydrazine HCl salt (68.7 g, 307 mmol) in portions. The mixture was heated to 85 °C and stirred for 2 h, cooled to rt, then 4 M HCl in 1,4-dioxane (69.8 mL, 279 mmol) added dropwise. The mixture was heated to 85 °C and stirred for an additional 3 h, then concentrated under reduced pressure and the residue was dissolved in TFA (0.78 L). The mixture was heated to 60 °C and stirred for 1.5 h, concentrated under reduced pressure, and the residue adjusted to pH ~5 with saturated NaHCO 3 , then extracted with EtOAc (3 × 1.5 L). The combined organic layers were dried over anhydrous Na 2 SO 4 , filtered, the filtrate concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give 3-(5-bromo-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-1H-indol-3-yl)-2,2-dimethylpropanoic acid and ethyl (S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropanoate (78 g, crude). LCMS (ESI): m/z [M+H] calculated for C21H23BrN2O3 430.1 and C23H27BrN2O3 458.1; found 431.1 and 459.1.

[00265] Step 3. To a mixture of 3-(5-bromo-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-1H-indol-3-yl)-2,2-dimethylpropanoic acid and ethyl (S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropanoate (198 g, 459 mmol) in DMF (1.8 L) at 0 °C under a N2 atmosphere was added Cs2CO3 (449 g, 1.38 mol) in portions. EtI (215 g, 1.38 mmol) in DMF (200 mL) was then added dropwise at 0 °C. The mixture was warmed to rt and stirred for 4 h and then diluted with brine (5 L) and extracted with EtOAc (3 × 2.5 L). The combined organic layers were washed with brine (2 × 1.5 L), dried over anhydrous Na 2 SO 4 , and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give ethyl 3-(5-bromo-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-3-yl)-2,2-dimethylpropanoate (160 g, 57% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C 25H 31 BrN 2 O 3 486.2; found 487.2.

[00266] Step 4. To a mixture of ethyl 3-(5-bromo-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-3-yl)-2,2-dimethylpropanoate (160 g, 328 mmol) in THF (1.6 L) at 0 °C under a N2 atmosphere was added LiBH4 (28.6 g, 1.3 mol). The mixture was heated at 60 °C for 16 h, cooled, and quenched with pre-cooled (0 °C) aqueous NH4Cl (5 L). The mixture was extracted with EtOAc (3 x 2 L), and the combined organic layers were washed with brine (2 x 1 L), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give two atropisomers (as single atropisomers) of 3-(5-bromo-1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropan-1-ol (60 g, 38% yield) and (40 g, 26% yield) both as solids. LCMS (ESI): m/z [M+H] calcd for C23H29BrN2O2 444.1; found 445.2. INTERMEDIATE 2 AND INTERMEDIATE 4. SYNTHESIS OF (S)-1-((S)-2-((TERT-BUTOXYCARBONYL)AMINO)-3-(3-(4,4,5,5-TETRAMETHYL-1,3,2-DIOXABOROLAN-2-YL)-5-((TRI-ISOPROPYLSILYL)OXY)PHENYL)PROPANOYL)HEXA-HYDROPYRIDAZINE-3-CARBOXYLATE

[00267] Step 1. To a mixture of (S)-methyl 2-(tert-butoxycarbonylamino)-3-(3-hydroxyphenyl)propanoate (10.0 g, 33.9 mmol) in DCM (100 mL) was added imidazole (4.6 g, 67.8 mmol) and TIPSCl (7.8 g, 40.7 mmol). The mixture was stirred at rt overnight and then diluted with DCM (200 mL) and washed with H2O (150 mL × 3). The organic layer was dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give (S)-methyl 2-tert-butoxycarbonylamino)-3-(3-(triisopropylsilyloxy)phenyl)-propanoate (15 g, 98% yield) as an oil. LCMS (ESI): m/z [M+Na] calculated for C24H41NO5SiNa 474.3; found 474.2.

[00268] Step 2. A mixture of (S)-methyl 2-(tert-butoxycarbonylamino)-3-(3-(triisopropylsilyloxy)phenyl)-propanoate (7.5 g, 16.6 mmol), PinB2 (6.3 g, 24.9 mmol), [Ir(OMe)(COD)]2 (1.1 g, 1.7 mmol), and 4-tert-butyl-2-(4-tert-butyl-2-pyridyl)pyridine (1.3 g, 5.0 mmol) was purged with Ar (x3), then THF (75 mL) was added and the mixture was placed under an atmosphere of Ar and sealing. The mixture was heated to 80 °C and stirred for 16 h, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give (S)-methyl 2-(tert-butoxycarbonylamino)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-(triisopropylsilyloxy)phenyl)-propanoate (7.5 g, 78% yield) as a solid. LCMS (ESI): m/z [M+Na] calcd for C30H52BNO7SiNa 600.4; found 600.4; 1H NMR (300 MHz, CD3OD) δ 7.18 (s, 1H), 7.11 (s, 1H), 6.85 (s, 1H), 4.34 (m, 1H), 3.68 (s, 3H), 3.08 (m, 1H), 2.86 (m, 1H), 1.41 - 1.20 (m, 26H), 1.20 - 1.01 (m, 22H), 0.98 - 0.79 (m, 4H).

[00269] Step 3. To a mixture of triisopropylsilyl (S)-2-((tert-butoxycarbonyl)amino)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-((triisopropylsilyl)oxy)phenyl)propanoate (4.95 g, 6.9 mmol) in MeOH (53 mL) at 0 °C was added LiOH (840 mg, 34.4 mmol) in H 2 O (35 mL). The mixture was stirred at 0 °C for 2 h, then acidified to pH ~5 with 1 M HCl and extracted with EtOAc (250 mL × 2). The combined organic layers were washed with brine (100 mL × 3), dried over anhydrous Na 2 SO 4 , filtered, and the filtrate concentrated under reduced pressure to give (S)-2-((tert-butoxycarbonyl)amino)-3-(3-(4,4,5,5-tetramethyl- 1,3,2-dioxaborolan-2-yl)-5-((triisopropylsilyl)oxy)phenyl)propanoic acid (3.7 g, 95% yield), which was used directly in the next step without further purification. LCMS (ESI): m/z [M+NH 4 ] calcd for C 29H 50BNO 7 SiNH 4 581.4; found 581.4.

[00270] Step 4. To a mixture of methyl (S)-hexahydropyridazine-3-carboxylate (6.48 g, 45.0 mmol) in DCM (200 mL) at 0 °C was added NMM (41.0 g, 405 mmol), (S)-2-((tert-butoxycarbonyl)amino)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-((triisopropylsilyl)oxy)phenyl)propanoic acid (24 g, 42.6 mmol) in DCM (50 mL) then HOBt (1.21 g, 9.0 mmol) and EDCI HCl salt (12.9 g, 67.6 mmol). The mixture was warmed to rt and stirred for 16 h, then diluted with DCM (200 mL) and washed with H2O (3 × 150 mL). The organic layer was dried over anhydrous Na2SO4, filtered, the filtrate concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give methyl (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-((triisopropylsilyl)oxy)phenyl)propanoyl)hexahydropyridazine-3-carboxylate (22 g, 71% yield) as an oil. LCMS (ESI): m/z [M+H] calculated for C35H60BN3O8Si 689.4; found 690.5.INTERMEDIATE 3. SYNTHESIS OF N-((S)-1-ACRYLOYLPYRROLIDINE-3-CARBONYL)-N-METHYL-L-VALINE

[00271] Step 1. To a mixture of (S)-1-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid (2.2 g, 10.2 mmol) in DMF (10 mL) at rt was added HATU (7.8 g, 20.4 mmol) and DIPEA (5 mL). After stirring at rt for 10 min, tert-butylmethyl-L-valinate (3.8 g, 20.4 mmol) in DMF (10 mL) was added. The mixture was stirred at rt for 3 h, then diluted with DCM (40 mL) and H2O (30 mL). The aqueous and organic layers were separated, and the organic layer was washed with H2O (3 × 30 mL), brine (30 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give (S)-tert-butyl 3-(((S)-1-(tert-butoxy)-3-methyl-1-oxobutan-2-yl(methyl)carbamoyl)pyrrolidine-1-carboxylate (3.2 g, 82% yield) as an oil. LCMS (ESI): m/z [M+Na] calcd for C20H36N2O5Na 407.3; found 407.2.

[00272] Step 2. A mixture of (S)-tert-butyl 3-(((S)-1-(tert-butoxy)-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)pyrrolidine-1-carboxylate (3.2 g, 8.4 mmol) in DCM (13 mL) and TFA (1.05 g, 9.2 mmol) was stirred at rt for 5 h. The mixture was concentrated under reduced pressure to give (S)-tert-butyl 3-methyl-2-((S)-N-methylpyrrolidine-3-carboxamido)butanoate (2.0 g, 84% yield) as an oil. LCMS (ESI): m/z [M+H] calcd for C15H28N2O3 284.2; found 285.2.

[00273] Step 3. To a mixture of (S)-tert-butyl 3-methyl-2-((S)-N-methylpyrrolidine-3-carboxamido)butanoate (600 mg, 2.1 mmol) in DCM (6 mL) at 0 °C was added TEA (342 mg, 3.36 mmol). After stirring at 0 °C for 10 min, acryloyl chloride (284 mg, 3.2 mmol) in DCM (10 mL) was added. The mixture was warmed to rt and stirred for 24 h, then diluted with DCM (30 mL) and H2O (30 mL). The aqueous and organic layers were separated, and the organic layer was washed with H2O (3 × 30 mL), brine (30 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give tert-butyl N-((S)-1-acryloylpyrrolidine-3-carbonyl)-N-methyl-L-valinate (500 mg, 70% yield) as an oil.

[00274] Step 4. To a mixture of tert-butyl N-((S)-1-acryloylpyrrolidine-3-carbonyl)-N-methyl-L-valinate (100 mg, 0.29 mmol) in DCM (3.0 mL) at 15 °C was added TFA (0.3 mL). The mixture was warmed to rt and stirred for 5 h, then the mixture was concentrated under reduced pressure to give N-((S)-1-acryloylpyrrolidine-3-carbonyl)-N-methyl-L-valine (150 mg) as a solid. The crude product was used directly in the next step without further purification. LCMS (ESI): m/z [M+H] calcd for C14H22N2O4 282.2; found 283.2.INTERMEDIATE 5. SYNTHESIS OF TERC-BUTYL ((63S,4S)-11 -ETHYL-12-(2-((S)-1 -METHOXYETHYL)PYRIDIN-3-YL)-10,10-DIMETHYL-5,7-DIOXO-25-((TRI-ISOPROPYLSYL)OXY)-61 ,62,63,64,65,66-HEXA-HYDRO-11H-8-OXA-1(5,3)-INDOLE-6(1,3)-PYRIDAZINE-2(1,3)-BENZENOCYCLOUNDECAPHANE-4-YL)CARBAMATE.

[00275] Step 1. To a stirred mixture of 3-(5-bromo-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-3-yl)-2,2-dimethylpropan-1-ol (30 g, 67 mmol) and methyl in 1,4-dioxane (750 mL) at rt under an Ar atmosphere was added Na2CO3 (17.9 g, English: 168.4 mmol), Pd(DtBPF)Cl2 (4.39 g, 6.7 mmol), and H2O (150.00 mL) in portions. The mixture was heated to 85 °C and stirred for 3 h, cooled, diluted with H2O (2 L), and extracted with EtOAc (3 × 1 L). The combined organic layers were washed with brine (2 × 500 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain methyl (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-5-yl]-5-[(triisopropylsilyl)oxy]phenyl]propanoyl]-1,2-diazinane-3-carboxylate (50 g, 72% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C52H77N5O8Si 927.6; found 928.8.

[00276] Step 2. To a stirred mixture of methyl (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-5-yl]-5-[(triisopropylsilyl)oxy]phenyl]propanoyl]-1,2-diazinane-3-carboxylate (50 g, 54 mmol) in DCE (500 mL) at rt was added trimethyltin hydroxide (48.7 g, 269 mmol) portionwise. The mixture was heated to 65 °C and stirred for 16 h, then filtered and the filter cake washed with DCM (3 × 150 mL). The filtrate was concentrated under reduced pressure to give (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-5-yl]-5-[(triisopropylsilyl)oxy]phenyl]propanoyl]-1,2-diazinane-3-carboxylic acid (70 g, crude), which was used directly in the next step without further purification. LCMS (ESI): m/z [M+H] calcd for C51H75N5O8Si 913.5; found 914.6.

[00277] Step 3. To a stirred mixture of (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-5-yl]-5-[(triisopropylsilyl)oxy]phenyl]propanoyl]-1,2-diazinane-3-carboxylic acid (70 g) in DCM (5 L) at 0 °C under a N2 atmosphere was added DIPEA (297 g, 2.3 mol), HOBT (51.7 g, 383 mmol), and EDCI (411 g, 2.1 mol) in portions. The mixture was warmed to rt and stirred for 16 h, then diluted with DCM (1 L), washed with brine (3 × 1 L), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give tert-butyl((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-25-((triisopropylsilyl)oxy)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (36 g, 42% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C51H73N5O7Si 895.5; found 896.5.INTERMEDIATE 6. SYNTHESIS OF tert-BUTYL N-[(8S,14S)-21-IODO-18,18-DIMETHYL- 9,15-DIOXO-4-[(TRI-ISOPROPYLSILYL)OXY]-16-OXA-10,22,28- TRIAZAPENTACYCLO[18.5.2.1A[2,6].1A[10,14].0A[23,27]]NONACOSA- 1(26),2,4,6(29),20,23(27),24-HEPTAEN-8-YL]CARBAMATE.

[00278] Step 1. This reaction was performed in 5 parallel batches on the scale illustrated below.

[00279] In 2 L round bottom flasks, 5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-1H-indole (100 g, 192 mmol) and TBAF (301.4 g, 1.15 mol) in THF (1.15 L) were each added at rt. The resulting mixture was heated to 50 °C and stirred for 16 h, then the mixture was concentrated under reduced pressure. The combined residues were diluted with H2O (5 L) and extracted with EtOAc (3 × 2 L). The combined organic layers were washed with brine (2 × 1.5 L), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give 3-(5-bromo-1H-indol-3-yl)-2,2-dimethylpropan-1-ol (310 g, crude) as a solid. LCMS (ESI): m/z [M+H] calculated for C13H16BrNO 281.0 and 283.0; found 282.1 and 284.1.

[00280] Step 2. This reaction was performed in two parallel batches on the scale illustrated below.

[00281] To a stirred mixture of 3-(5-bromo-1H-indol-3-yl)-2,2-dimethylpropan-1-ol (135 g, 478 mmol) and TEA (145.2 g, 1.44 mol) in DCM (1.3 L) at 0 °C under a N2 atmosphere was added Ac2O (73.3 g, 718 mmol) and DMAP (4.68 g, 38.3 mmol) in portions. The resulting mixture was stirred for 10 min at 0 °C, then washed with H2O (3 × 2 L). The organic layers from each experiment were combined and washed with brine (2 × 1 L), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 3-(5-bromo-1H-indol-3-yl)-2,2-dimethylpropyl acetate (304 g, 88% yield) as a solid. 1H NMR (400 MHz, DMSO-d6) δ 11.16–11.11 (m, 1H), 7.69 (d, J = 2.0 Hz, 1H), 7.32 (d, J = 8.6 Hz, 1H), 7.19–7.12 (m, 2H), 3.69 (s, 2H), 2.64 (s, 2H), 2.09 (s, 3H), 0.90 (s, 6H).

[00282] Step 3. This reaction was performed in four parallel batches on the scale illustrated below.

[00283] In 2 L round bottom flasks were added methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-[(triisopropylsilyl)oxy]phenyl]propanoate (125 g, 216 mmol), 1,4-dioxane (1 L), H2O (200 mL), 3-(5-bromo-1H-indol-3-yl)-2,2-dimethylpropyl acetate (73.7 g, 227 mmol), K2CO3 (59.8 g, 433 mmol), and Pd(DtBPF)Cl2 (7.05 g, 10.8 mmol) at rt under an Ar atmosphere. The resulting mixture was heated to 65 °C and stirred for 2 h, then diluted with H2O (10 L) and extracted with EtOAc (3 × 3 L). The combined organic layers were washed with brine (2 × 2 L), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain methyl (2S)-3-(3-[3-[3-(acetyloxy)-2,2-dimethylpropyl]-1H-indol-5-yl]-5-[(triisopropylsilyl)oxy]phenyl)-2-[(tert-butoxycarbonyl)amino]propanoate (500 g, 74% yield) as an oil. LCMS (ESI): m/z [M+Na] calcd for C39H58N2O7SiNa 717.4; found 717.3.

[00284] Step 4. This reaction was performed in three parallel batches on the scale illustrated below.

[00285] To a stirred mixture of methyl (2S)-3-(3-[3-[3-(acetyloxy)-2,2-dimethylpropyl]-1H-indol-5-yl]-5-[(triisopropylsilyl)oxy]phenyl)-2-[(tert-butoxycarbonyl)amino]propanoate (150 g, 216 mmol) and NaHCO3 (21.76 g, 259 mmol) in THF (1.5 L) was added AgOTf (66.5 g, 259 mmol) in THF dropwise at 0 °C under a nitrogen atmosphere. I2 (49.3 g, 194 mmol) in THF was added dropwise over 1 h at 0 °C, and the resulting mixture was stirred for an additional 10 min at 0 °C. The combined experiments were diluted with aqueous Na2S2O3 (5 L) and extracted with EtOAc (3 × 3 L). The combined organic layers were washed with brine (2 × 1.5 L), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain methyl (2S)-3-(3-[3-[3-(acetyloxy)-2,2-dimethylpropyl]-2-iodo-1H-indol-5-yl]-5-[(triisopropylsilyl)oxy]phenyl)-2-[(tert-butoxycarbonyl)amino]propanoate (420 g, 71% yield) as an oil. LCMS (ESI): m/z [M+Na] calculated for C39H57IN2O7SiNa, 843.3; found 842.9.

[00286] Step 5. This reaction was performed in three parallel batches on the scale illustrated below.

[00287] To a 2 L round bottom flask were added methyl (2S)-3-(3-[3-[3-(acetyloxy)-2,2-dimethylpropyl]-2-iodo-1H-indol-5-yl]-5-[(triisopropylsilyl)oxy]phenyl)-2-[(tert-butoxycarbonyl)amino]propanoate (140 g, 171 mmol), MeOH (1.4 L), and K3PO4 (108.6 g, 512 mmol) at 0 °C. The mixture was warmed to rt and stirred for 1 h, then the combined experiments were diluted with H2O (9 L) and extracted with EtOAc (3 × 3 L). The combined organic layers were washed with brine (2 × 2 L), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to give methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl]-5-[(triisopropylsilyl)oxy]phenyl]propanoate (438 g, crude) as a solid. LCMS (ESI): m/z [M+Na] calcd for C37H55IN2O6SiNa 801.3; found 801.6.

[00288] Step 6. This reaction was performed in three parallel batches on the scale illustrated below.

[00289] To a stirred mixture of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl]-5-[(triisopropylsilyl)oxy]phenyl]propanoate (146 g, 188 mmol) in THF (1.46 L) was added LiOH (22.45 g, 937 mmol) in H2O (937 mL) dropwise at 0 °C. The resulting mixture was warmed to rt and stirred for 1.5 h [note: LCMS showed 15% de-TIPS product]. The mixture was acidified to pH 5 with 1 M HCl (1 M) and the combined experiments were extracted with EtOAc (3 × 3 L). The combined organic layers were washed with brine (2 × 2 L), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to give (2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl]-5-[(triisopropylsilyl)oxy]phenyl]propanoic acid (402 g, crude) as a solid. LCMS (ESI): m/z [M+Na] calcd for C36H53IN2O6SiNa 787.3; found 787.6.

[00290] Step 7. To a stirred mixture of (2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl]-5-[(triisopropylsilyl)oxy]phenyl]propanoic acid (340 g, 445 mmol) and methyl (3S)-1,2-diazinane-3-carboxylate (96.1 g, 667 mmol) in DCM (3.5 L) was added NMM (225 g, 2.2 mol), EDCI (170 g, 889 mmol), and HOBT (12.0 g, 88.9 mmol) in portions at 0 °C. The mixture was warmed to rt and stirred for 16 h, then washed with H2O (3 × 2.5 L), brine (2 × 1 L), dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain methyl (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl]-5-[(triisopropylsilyl)oxy]phenyl]propanoyl]-1,2-diazinane-3-carboxylate (310 g, 62% yield) as an oil. LCMS (ESI): m/z [M+H] calcd for C42H63IN4O7Si 890.4; found 890.8.

[00291] Step 8. This reaction was performed in three parallel batches on the scale illustrated below.

[00292] To a stirred mixture of methyl (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl]-5-[(triisopropylsilyl)oxy]phenyl]propanoyl]-1,2-diazinane-3-carboxylate (85.0 g, 95.4 mmol) in THF (850 mL) was each added LiOH (6.85 g, 286 mmol) in H2O (410 mL) dropwise at 0 °C under an atmosphere of N2. The mixture was stirred at 0 °C for 1.5 h [note: LCMS showed 15% de-TIPS product], then acidified to pH 5 with 1 M HCl and the combined experiments extracted with EtOAc (3 × 2 L). The combined organic layers were washed with brine (2 × 1.5 L), dried over anhydrous Na 2 SO 4 , filtered, and the filtrate was concentrated under reduced pressure to give (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl]-5-[(triisopropylsilyl)oxy]phenyl]propanoyl]-1,2-diazinane-3-carboxylic acid (240 g, crude) as a solid. LCMS (ESI): m/z [M+H] calculated for C41H61IN4O7Si 876.3; found 877.6.

[00293] Step 9. This reaction was performed in two parallel batches on the scale illustrated below.

[00294] To a stirred mixture of (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl]-5-[(triisopropylsilyl)oxy]phenyl]propanoyl]-1,2-diazinane-3-carboxylic acid (120 g, 137 mmol) in DCM (6 L) was added DIPEA (265 g, 2.05 mol), EDCI (394 g, 2.05 mol), and HOBT (37 g, 274 mmol) in portions at 0 °C under a N2 atmosphere. The mixture was warmed to rt and stirred overnight, then the combined experiments were washed with H2O (3 × 6 L), brine (2 × 6 L), dried over anhydrous Na2SO4, and filtered. After filtration, the filtrate was concentrated under reduced pressure. The filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to give tert-butyl N-[(8S,14S)-21-iodo-18,18-dimethyl-9,15-dioxo-4-[(triisopropylsilyl)oxy]-16-oxa-10,22,28-triazapentacyclo[18.5.2.1A[2,6]. 1 A[10,14].0A[23,27]]nonacosa- 1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamate (140 g, 50% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C41H59IN4O6Si 858.9; found 858.3.INTERMEDIATE 7. SYNTHESIS OF (S)-METHYL 2-((TERT-BUTOXYCARBONYL)AMINθ)-3-(4-(4,4,5,5-TETRAMETHYL-1,3,2-DIOXABOROLAN-2-YL)PYRIDIN-2-YL)PROPANOATE

[00295] Step 1. Zn powder (28 g, 428 mmol) was added to a 1 L three-necked round bottom flask, purged with N2, and heated with a heat gun for 10 min under vacuum. The mixture was cooled to rt and a solution of 1,2-dibromoethane (1.85 mL, 21.5 mmol) in DMF (90 mL) was added dropwise over 10 min. The mixture was heated at 90 °C for 30 min and cooled back to rt. English: TMSCl (0.55 mL, 4.3 mmol) was added and the mixture was stirred for 30 min at rt, then a mixture of (R)-methyl 2-((tert-butoxycarbonyl)amino)-3-iodopropanoate (22.5 g, 71.4 mmol) in DMF (200 mL) was added dropwise over a period of 10 min. The mixture was heated to 35 °C and stirred for 2 h, then cooled to rt and 2,4-dichloropyridine (16 g, 109 mmol) and Pd(PPh3)2Cl2 (4 g, 5.7 mmol) were added. The mixture was heated to 45 °C and stirred for 2 h, cooled and filtered, then H2O (1 L) and EtOAc (0.5 L) were added to the filtrate. The organic and aqueous layers were separated, and the aqueous layer was extracted with EtOAc (2 × 500 mL). The organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude residue was purified by silica gel column chromatography to give (S)-methyl 2-((tert-butoxycarbonyl)amino)-3-(4-chloropyridin-2-yl)propanoate (6.5 g, 29% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C14H19ClN2O4 314.1; found 315.1.

[00296] Step 2. To a mixture of (S)-methyl 2-((tert-butoxycarbonyl)amino)-3-(4-chloropyridin-2-yl)propanoate (6.5 g, 20.6 mmol) in 1,4-dioxane (80 mL) at rt under a N2 atmosphere was added bis(pinacolato)diboron (6.3 g, 24.7 mmol), KOAc (8.1 g, 82.4 mmol), and Pd(PCy3)2Cl2 (1.9 g, 2.5 mmol). The mixture was heated to 100 °C and stirred for 3 h, then H2O (100 mL) was added and the mixture was extracted with EtOAc (3 × 200 mL). The organic layers were combined, washed with brine (2 × 100 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give (S)-methyl 2-((tert-butoxycarbonyl)amino)-3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)propanoate (6 g, 72% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C20H31BN2O6 406.2; found 407.3. SYNTHESIS OF INTERMEDIATE 8.

[00297] Step 1. To a mixture of 4-(dimethylamino)but-2-ynoic acid (900 mg, 7.0 mmol) in DMF (20 mL) at -5 °C was added tert-butyl N-methyl-N-((S)-pyrrolidine-3-carbonyl)-L-valinate (1.0 g, 3.5 mmol), DIPEA (2.2 g, 17.6 mmol), and HATU (2.7 g, 7.0 mmol) in portions. The mixture was stirred at -5 to 5 °C for 1 h, then diluted with EtOAc (100 mL) and ice-H2O (100 mL). The aqueous and organic layers were separated, and the organic layer was washed with H2O (3 × 100 mL), brine (100 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give tert-butyl N-((S)-1-(4-(dimethylamino)but-2-ynoyl)pyrrolidine-3-carbonyl)-N-methyl-L-valinate (900 mg, 55% yield) as an oil. LCMS (ESI): m/z [M+H] calcd for C21H35N3O4 393.5; found 394.3.

[00298] Step 2. To a mixture of tert-butyl N-((S)-1-(4-(dimethylamino)but-2-ynoyl)pyrrolidine-3-carbonyl)-N-methyl-L-valinate (260 mg, 0.66 mmol) in DCM (6 mL) was added TFA (3 mL) at rt. The mixture was stirred at rt for 2 h, then the solvent was concentrated under reduced pressure to give (2S)-2-{1-[(3S)-1-[4-(dimethylamino)but-2-ynoyl]pyrrolidin-3-yl]-N-methylformamido}-3-methylbutanoic acid (280 mg) as an impure oil. The crude product was used directly in the next step without further purification. LCMS (ESI): m/z [M+H] calcd for C17H27N3O4 337.2; found 338.3.INTERMEDIATE SYNTHESIS 9.

[00299] Step 1. To a mixture of tert-butyl N-methyl-N-((S)-pyrrolidine-3-carbonyl)-L-valinate (500 mg, 1.8 mmol) in DCM (8 mL) at 5 °C was added TEA (533 mg, 5.3 mmol) followed by dropwise addition of 2-chloroethane-1-sulfonyl chloride (574 mg, 3.5 mmol) in DCM (2 mL). The mixture was stirred at 5 °C for 1 h, then diluted with H2O (20 mL) and extracted with EtOAC (3 × 10 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain tert-butyl N-methyl-N-((S)-1-(vinylsulfonyl)pyrrolidine-3-carbonyl)-L-valinate (300 mg, 45% yield) as an oil. LCMS (ESI): m/z [M+H] calcd for C17H30N2O5S 374.2; found 375.2.

[00300] Step 2. To a mixture of tert-butyl N-methyl-N-((S)-1-(vinylsulfonyl)pyrrolidine-3-carbonyl)-L-valinate (123 mg, 0.33 mmol) in DCM (3 mL) at rt was added TFA (1 mL). The mixture was stirred at rt for 1 h, then concentrated under reduced pressure to give N-methyl-N-((S)-1-(vinylsulfonyl)pyrrolidine-3-carbonyl)-L-valine (130 mg, crude) as a solid, which was used directly in the next step without further purification. LCMS (ESI): m/z [M+H] calcd for C13H22N2O5S 318.1; found 319.1. SYNTHESIS OF INTERMEDIATE 10.

[00301] Step 1. A mixture of 5-chloro-1H-pyrrolo[3,2-b]pyridine-3-carbaldehyde (8.5 g, 47.1 mmol) and ethyl 2-(triphenylphosphoranylidene)propionate (2.56 g, 70.7 mmol) in 1,4-dioxane (120 mL) was stirred at reflux for 4 h, then concentrated under reduced pressure. EtOAc (200 mL) was added and the mixture was washed with brine, dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give (E)-3-(5-chloro-1H-pyrrolo[3,2-b]pyridin-3-yl)-2-methylacrylate (7.5 g, 60% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C13H13ClN2O2 264.1; found 265.1.

[00302] Step 2. To a mixture of ethyl (E)-3-(5-chloro-1H-pyrrolo[3,2-b]pyridin-3-yl)-2-methylacrylate (7.5 g, 28.3 mmol) and NiCl2 (4.8 g, 28.3 mmol) in 1:1 THF/MeOH (300 mL) was added NaBH4 (21.5 g, 566 mmol) in 20 portions every 25 min. After complete addition, the mixture was stirred at rt for 30 min, then diluted with EtOAc (500 mL) and washed with brine, dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give ethyl 3-(5-chloro-1H-pyrrolo[3,2-b]pyridin-3-yl)-2-methylpropanoate (3.4 g, 45% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C13H15ClN2O2 266.1; found 267.1.

[00303] Step 3. To a mixture of ethyl 3-(5-chloro-1H-pyrrolo[3,2-b]pyridin-3-yl)-2-methylpropanoate (7.0 g, 26.2 mmol) and AgOTf (6.7 g, 26.2 mmol) in THF (50 mL) at 0 °C was added I2 (6.65 g, 26.2 mol). The mixture was stirred at 0 °C for 30 min, then diluted with EtOAc (100 mL), washed with Na2SO3 (50 mL), brine (50 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give ethyl 3-(5-chloro-2-iodo-1H-pyrrolo[3,2-b]pyridin-3-yl)-2-methylpropanoate (6 g, 58% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C13H14ClIN2O2 392.0; found 393.0.

[00304] Step 4. To a mixture of ethyl 3-(5-chloro-2-iodo-1H-pyrrolo[3,2-b]pyridin-3-yl)-2-methylpropanoate (6.0 g, 15.3 mmol) and 2-(2-(2-methoxyethyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (5.6 g, 21.4 mmol) and K2CO3 (6.3 g, 45.9 mmol) in 1,4-dioxane (150 mL) and H2O (30 mL) under a N2 atmosphere was added Pd(dppf)Cl2.DCM (1.3 g, 3.1 mmol). The mixture was heated to 80 °C and stirred for 4 h, then diluted with EtOAc (500 mL), washed with brine, dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 3-(5-chloro-2-(2-(2-methoxyethyl)phenyl)-1H-pyrrolo[3,2-b]pyridin-3-yl)-2-methylpropanoate (5.5 g, 50% yield) as a viscous oil. LCMS (ESI): m/z [M+H] calcd for C22H25ClN2O3 400.2; found 401.2.

[00305] Step 5. A mixture of ethyl 3-(5-chloro-2-(2-(2-methoxyethyl)phenyl)-1H-pyrrolo[3,2-b]pyridin-3-yl)-2-methylpropanoate (5.5 g, 13.8 mmol), Cs2CO3 (8.9 g, 27.5 mmol), and EtI (3.5 g, 27.5 mmol) in DMF (30 mL) at rt was stirred for 10 h. The mixture was diluted with EtOAc (100 mL), washed with brine (20 mL x 4), dried over Na2SO4, filtered, and concentrated in vacuo. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give 3-(5-chloro-1-ethyl-2-(2-(2-methoxyethyl)phenyl)-1H-pyrrolo[3,2-b]pyridin-3-yl)-2-methylpropanoate (5.6 g, 95% yield) as a viscous oil. LCMS (ESI): m/z [M+H] calcd for C25H31ClN2O3 428.2; found 429.2.

[00306] Step 6. To a mixture of ethyl 3-(5-chloro-1-ethyl-2-(2-(2-methoxyethyl)phenyl)-1H-pyrrolo[3,2-b]pyridin-3-yl)-2-methylpropanoate (5.4 g, 12.6 mmol) in THF (50 mL) at -65 °C was added 2M LDA (25 mL, 50 mmol) and stirred at -65 °C for 1 h. MeI (3.6 g, 25 mmol) was added and the mixture was stirred at -65 °C for 2.5 h, then aqueous NH4Cl and EtOAc (50 mL) were added. The aqueous and organic layers were separated and the organic layer was washed with brine (30 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give ethyl 3-(5-chloro-1-ethyl-2-(2-(2-methoxyethyl)phenyl)-1H-pyrrolo[3,2-b]pyridin-3-yl)-2,2-dimethylpropanoate (3.2 g, 57% yield) as a viscous oil. LCMS (ESI): m/z [M+H] calcd for C25H31ClN2O3 442.2; found 443.2.

[00307] Step 7. To a mixture of ethyl 3-(5-chloro-1-ethyl-2-(2-(2-methoxyethyl)phenyl)-1H-pyrrolo[3,2-b]pyridin-3-yl)-2,2-dimethylpropanoate (1.0 g, 2.3 mmol) in THF (10 mL) at 5 °C was added LiBH4 (196 mg, 9.0 mmol). The mixture was heated to 65 °C and stirred for 2 h, then NH4Cl and EtOAc (50 mL) were added. The aqueous and organic layers were separated, and the organic layer was washed with brine (30 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give 3-(5-chloro-1-ethyl-2-(2-(2-methoxyethyl)phenyl)-1H-pyrrolo[3,2-b]pyridin-3-yl)-2,2-dimethylpropan-1-ol (0.75 g, 82% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C23H29ClN2O2 400.2; found 401.2.INTERMEDIATE 11: (3S)-1-{(2S)-2-(TERT-BUTOXYCARBONYL)AMINO-3-[3-FLUORO-5-(4,4,5,5-TETRAMETHYL-1,3,2-DIOXABOROLAN-2-YL)PHENYL]PROPANOYL}-1,2-DIAZINAN-3-CARBOXYLATE

[00308] Step 1. To a stirred solution of methyl (2R)-2-{[(tert-butoxy)carbonyl]amino}-3-(iodozincium)propanoate (12 g, 30 mmol, 1.2 eq) in DMF (100 mL) was added 1-bromo-3-fluoro-5-iodobenzene (7.5 g, 25 mmol, 1 eq) and Pd(PPh3)2Cl2 (1.7 g, 2.5 mmol, 0.1 equiv) at 20°C under N2 atmosphere. The resulting mixture was stirred for 2 h at 65°C under N2 atmosphere. The reaction mixture was quenched with water and extracted with EA (200 mL x 2). The organic phase was washed with water (200 mL x 1) and brine (100 mL x 1) and concentrated to dryness to give a residue. The residue was purified by preparative TLC (PE/EA=10/1) to give methyl 3-(3-bromo-5-fluorophenyl)-2-{[(tert-butoxy)carbonyl]amino}propanoate (6 g, 58% yield) as a colorless oil. LCMS (ESI) m/z = 398.1 [M+Na]+ , calcd for C15H19BrFNO4: 375.0

[00309] Step 2. To a solution of methyl 3-(3-bromo-5-fluorophenyl)-2-{[(tert-butoxy)carbonyl]amino}propanoate (3.2 g, 8.5 mmol, 1 eq) in THF (50 mL) was added lithium hydroxide (610.7 mg, 25.5 mmol, 3 eq) in H2O (10 mL). Then, the reaction mixture was stirred at 20 °C for 1 h. The mixture was adjusted to pH = 5.0 with 1 M aqueous HCl solution. The mixture was quenched with H2O (150 mL) and extracted with EA (200 mL x 3). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, and concentrated to give 3-(3-bromo-5-fluorophenyl)-2-{[(tert-butoxy)carbonyl]amino}propanoic acid (2.65 g, 68% yield) as a white solid. LCMS (ESI) m/z = 384.1 [M+Na]+, calcd for C14H15BrFNO4 MW: 361.0

[00310] Step 3. To a mixture of 3-(3-bromo-5-fluorophenyl)-2-{[(tert-butoxy)carbonyl]amino}propanoic acid (2.3 g, 6.4 mmol, 1 eq) and methyl (3S)-1,2-diazinane-3-carboxylate (1.66 g, 11.5 mmol, 1.8 eq) in DMF (150 mL) was added HATU (4.9 g, 12.8 mmol, 2 eq) and DIEA (16.5 g, 128 mmol, 20 eq) in DMF (50 mL) at 0 °C. Then, the reaction mixture was stirred at 0 °C for 1 h. The mixture was quenched with H2O (100 mL) and extracted with EA (300 mL x 3). The combined organic layers were washed (50 mL), dried over Na2SO4 and concentrated to give the residue, which was purified by Pre-HPLC eluting with 60% to 70% acetonitrile in water (0.1% FA) in 10 min to give methyl (3S)-1-[(2S)-3-(3-bromo-5-fluorophenyl)-2-{[(tert-butoxy)carbonyl]amino}propanoyl]-1,2-diazinane-3-carboxylate (2.7 g, 78% yield) as a pale yellow solid. LCMS (ESI) m/z = 510.1 [M+Na]+, calcd for C20H27BrFNO5: 487.1.

[00311] Step 4. A mixture of methyl (S)-1-((S)-3-(3-bromo-5-fluorophenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate (3 g, 6.16 mmol, 1 eq), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (1.9 g, 7.4 mmol, 1.2 eq), KOAc (900 mg, 9.24 mmol, 1.5 eq), and Pd(dppf)Cl2DCM (0.3 g, 0.37 mmol, 0.05 eq) in dioxane (50 mL) was heated at 100 °C for 17 h under N2 atmosphere. The mixture was concentrated and purified by column chromatography (DCM/MeOH=100/1 to 40/1) to obtain methyl (3S)-1-(2S)-2-{(tert-butoxycarbonyl)amino-3-[3-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]propanoyl}-1,2-diazinane-3-carboxylate (2.6 g, 79% yield) as a yellow oil. LCMS (ESI) m/z = 536.2 [M+H]+, calcd for C26H39BFNO7: 535.3.

[00312] Compounds A341 and A342 can be prepared using methods disclosed herein via Intermediate 11. EXAMPLE A75. SYNTHESIS OF TWO ATROPISOMERS OF (2S)-N-[(8S,14S,20M)- 22-ETHYL-4-HYDROXY-21-{2-[(1S)-1-METHOXYETHYL]PYRIDIN-3-YL}-18,18-DIMETHYL-9,15-DIOXO-16-OXA-10,22,28-TRIAZAPENTACYCLE[18.5.2.12,6.110,14.023,27] NONACOSA-1 (26)

[00313] Step 1. To a stirred mixture of tert-butyl((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-25-((triisopropylsilyl)oxy)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (18.0 g, 20.1 mmol) in THF (180 mL) at 0 °C was added a 1 M solution of TBAF in THF (24.1 mL, 24.1 mmol). The mixture was stirred at 0 °C for 1 h, then diluted with brine (1.5 L) and extracted with EtOAc (3 × 1 L). The combined organic layers were washed with brine (2 × 500 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give tert-butyl((63S,4S)-11-ethyl-25-hydroxy-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (11.5 g, 69% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C42H53N5O7 739.4; found 740.4.

[00314] Step 2. To a stirred mixture of tert-butyl ((63S,4S)-11-ethyl-25-hydroxy-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexa- hydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundephane-4-yl)carbamate (11.5 g, 15.5 mmol) in DCM (120 mL) at 0°C was added TFA (60 mL, 808 mmol). The mixture was stirred at 0 °C for 1 h, then concentrated under reduced pressure and the residue again concentrated under reduced pressure with toluene (20 mL; repeated ×3) to give (63S,4S)-4-amino-11-ethyl-25-hydroxy-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-5,7-dione (12 g, crude), which was used directly in the next step without further purification. LCMS (ESI): m/z [M+H] calcd for C37H45N5O5 639.3; found 640.6.

[00315] Step 3. To a stirred mixture of (63S,4S)-4-amino-11-ethyl-25-hydroxy-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-5,7-dione (11.9 g, 18.6 mmol) in DMF (240 mL) at 0 °C under an atmosphere of N 2 was added DIPEA (48.1 g, 372 mmol), (2S)-3-methyl-2-[N-methyl-1-[(3S)-1-(methyl ... English:[(2-[(2-[(2-[(2-[(2-[(2-[(2-[(2-( ... The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (× 2) to give two atropisomers of (2S)-N-[(8S,14S,20M)-22-ethyl-4-hydroxy-21-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonacosa-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]-3-methyl-2-{M-methyl-1-[(3S)-1 -(prop-2-enoyl)pyrrolidin-3-yl]formamido}butanamide (2.7 g, 15.5% yield) and (4.2 g, 24.7% yield) both as solids. LCMS (ESI): m/z [M+H] calculated for C51H65N7O8 903.5; found 904.7; 1H NMR (400 MHz, DMSO-d6) δ 9.35 - 9.27 (m, 1H), 8.77 (dd, J = 4.7, 1.7 Hz, 1H), 7.95 (dq, J = 6.2, 2.0 Hz, 2H), 7.55 (ddd, J = 28.0, 8.2, 4.3 Hz, 3H), 7.08 (dd, J = 37.9, 6.2 Hz, 2H), 6.69 - 6.48 (m, 2H), 6.17 (ddt, J = 16.7, 7.2, 2.3 Hz, 1H), 5.74 - 5.62 (m, 1H), 5.43 - 5.34 (m, 1H), 5.12 - 5.00 (m, 1H), 4.25 (d, J = 12.3 Hz, 1H), 4.17 - 3.99 (m, 3H), 3.89 - 3.65 (m, 4H), 3.66 - 3.45 (m, 3H), 3.12 (s, 4H), 2.95 - 2.70 (m, 6H), 2.41 - 2.06 (m, 5H), 1.99 - 1.88 (m, 1H), 1.82 (d, J = 12.1 Hz, 2H), 1.54 (t, J = 12.0 Hz, 1H), 1.21 (dd, J = 6.3, 2.5 Hz, 3H), 1.11 (t, J = 7.1 Hz, 3H), 0.99 - 0.88 (m, 6H), 0.79 (ddd, J = 27.8, 6.7, 2.1 Hz, 3H), 0.48 (d, J = 3.7 Hz, 3H) and LCMS (ESI): m/z [M+H] calculated for C51H65N7O8 903.5; found 904.7; 1H NMR (400 MHz, DMSO-d6) δ 9.34 - 9.27 (m, 1H), 8.77 (dd, J = 4.7, 1.7 Hz, 1H), 8.17 - 7.77 (m, 3H), 7.64 - 7.43 (m, 3H), 7.33 (d, J = 13.7 Hz, 1H), 7.05 - 6.94 (m, 1H), 6.69 - 6.41 (m, 2H), 6.26 - 5.94 (m, 1H), 5.73 - 5.63 (m, 1H), 5.50 - 5.20 (m, 2H), 4.40 - 4.15 (m, 3H), 4.00 - 3.40 (m, 9H), 3.11 (d, J = 4.4 Hz, 3H), 2.93 - 2.60 (m, 8H), 2.29 - 2.01 (m, 3H), 1.99 (s, 1H), 1.87 - 1.75 (m, 2H), 1.73 - 1.47 (m, 2H), 1.40 (d, J = 6.0 Hz, 3H), 1.01 - 0.88 (m, 6H), 0.85 - 0.65 (m, 7H), 0.56 (s, 3H).EXAMPLE A89. SYNTHESIS OF (2S)-N-[(8S,14S)-22-ETHYL-4-HYDROXY-18,18-DIMETHYL-21-[6-(4- METHYLPIPERAZIN-1-IL)PYRIDIN-3-IL|-9,15-DIOXO-16-OXA-10,22,28-TRIAZAPENTAacLO[18.5.2.12,6.110,14.023,27]NONACOSA-1(26),2,4,6(29),20,23(27),24- HEPTAEN-8-IL|-3-METHYL-2-{N-METHYL-1-[(3S)-1-(PROP-2-ENOYL)PIRROLIDIN-3- IL]FORMAMIDO}BUTANAMIDE.

[00316] Step 1. To a mixture of tert-butyl((63S,4S)-12-iodo-10,10-dimethyl-5,7-dioxo-25-((triisopropylsilyl)oxy)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (240.00 mg, 0.279 mmol, 1.00 equiv) and Cs2CO3 (182 mg, 0.558 mmol, 2 equiv) in DMF (5.00 mL) was added ethyl iodide (113.45 mg, 0.727 mmol, 2.60 equiv) dropwise dropwise at 0 °C. The reaction was stirred for 16 h at 25 °C. The resulting mixture was diluted with water (10 mL) and extracted with EtOAc (3 × 10 mL). The combined organic layers were washed with brine (3 × 10 mL) and dried over anhydrous Na2SO4. The filtrate was concentrated under reduced pressure, and the remaining residue was purified by silica gel column chromatography to give tert-butyl ((63S,4S)-11-ethyl-12-iodo-10,10-dimethyl-5,7-dioxo-25-((triisopropylsilyl)oxy)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenecycloundecaphane-4-yl)carbamate (190 mg, 77% yield) as a yellow solid.

[00317] Step 2. A mixture of tert-butyl ((63S,4S)-11-ethyl-12-iodo-10,10-dimethyl-5,7-dioxo-25-((tri-isopropylsilyl)oxy)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)- indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundephane-4-yl)carbamate(500 mg, 0.54 mmol), 1-methyl-4-[5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl]piperazine (257 mg, 0.8 mmol), Pd(dppf)Cl2 (83 mg, 0.11 mmol) and K2CO3 (156 mg, 1.1 mmol) in 1,4-dioxane (25 mL) and H2O (5 mL) under an Ar atmosphere was stirred at 80 C for 2 h. The mixture was concentrated under reduced pressure and the residue was purified by prep-TLC to give tert-butyl((63S,4S)-11-ethyl-10,10-dimethyl-12-(6-(4-methylpiperazin-1-yl)pyridin-3-yl)-5,7-dioxo-25-((triisopropylsilyl)oxy)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (400 mg, 76% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C53H77N7O6Si 935.6; found 936.6.

[00318] Step 3. A mixture of tert-butyl((63S,4S)-11-ethyl-10,10-dimethyl-12-(6-(4-methylpiperazin-1-yl)pyridin-3-yl)-5,7-dioxo-25-((triisopropylsilyl)oxy)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (350 mg, 0.36 mmol) and 1M TBAF in THF (0.4 mL, 0.4 mmol) in THF (5 mL) was stirred at 0 °C for 1 h. The mixture was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give tert-butyl((63S,4S)-11-ethyl-25-hydroxy-10,10-dimethyl-12-(6-(4-methylpiperazin-1-yl)pyridin-3-yl)-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (290 mg, 100% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C44H57N7O6 779.4; found 780.4.

[00319] Step 4. A mixture of tert-butyl ((63S,4S)-11-ethyl-25-hydroxy-10,10-dimethyl-12-(6-(4-methylpiperazin-1-yl)pyridin-3-yl)-5,7-dioxo-61,62,63,64,65,66-hexa- hydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundephane-4-yl)carbamate (300 mg, 0.37 mmol) in TFA (5 mL) and DCM (5 mL) was stirred at rt for 1 h. The mixture was concentrated under reduced pressure to give (63S,4S)-4-amino-11-ethyl-25-hydroxy-10,10-dimethyl-12-(6-(4-methylpiperazin-1-yl)pyridin-3-yl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-5,7-dione (300 mg, crude) as a solid. LCMS (ESI): m/z [M+H] calcd for C39H49N7O4 679.4; found 680.3.

[00320] Step 5. To a mixture of (63S,4S)-4-amino-11-ethyl-25-hydroxy-10,10-dimethyl-12-(6-(4-methylpiperazin-1-yl)pyridin-3-yl)-61,62,63,64,65,66-hexahydro- 11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundephane-5,7-dione (300 mg, 0.36 mmol) in DMF (3 mL) at 0 C under an N2 atmosphere was added DIPEA (0.96 mL, 5.4 mmol) and acid (2S)-3-methyl-2-[N-methyl-1-[(3S)-1-(prop-2- [[enoyl]pyrrolidin-3-yl]formamido]butanoic acid (213 mg, 0.72 mmol) was added, followed by dropwise addition of COMU (243 mg, 0.56 mmol). H2O was added at 0 °C, and the mixture was extracted with EtOAc (3 × 10 mL). The combined organic layers were washed with brine (3 × 10 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the crude residue was purified by Prep-HPLC to give (2S)-N-[(8S,14S)-22-ethyl-4-hydroxy-18,18-dimethyl-21-[6-(4-methylpiperazin-1-yl)pyridin-3-yl]-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14023,27]nonacosa- 1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]-3-methyl-2-{N-methyl-1-[(3S)-1-(prop-2-enoyl)pyrrolidin-3-yl]formamido}butanamide (45 mg, 13.2% yield) as a solid. LCMS (ESI): m/z [M+H] calculated C53H69N9O7 943.5; found 944.8; 1H NMR (400 MHz, DMSO-d6) δ 9.39 - 9.23 (m, 1H), 8.64 - 8.60 (m, 1H), 8.19 - 8.16 (m, 1H), 8.15 (d, J = 6.2 Hz, 1H), 7.86 (s, 1H), 7.66 - 7.62 (m, 1H), 7.56-7.54 (m, 1H), 7.50 - 7.43 (m, 1H), 7.13 - 7.11 (m, 1H), 7.03 - 6.95 (m, 1H), 6.70 - 6.47 (m, 2H), 6.17 (ddt, J = 16.8, 6.4, 2.8 Hz, 1H), 5.76 - 5.63 (m, 1H), 5.45 - 5.33 (m, 1H), 5.11 (m, 1H), 4.75 - 4.72 (m, 1H), 4.28 - 4.24 (m, 1H), 4.11 - 3.98 (m, 4H), 3.91 - 3.76 (m, 1H), 3.73 - 3.71 (m, 1H), 3.59 - 3.56 (m, 7H), 3.51 - 3.40 (m, 2H), 3.08 - 2.94 (m, 1H), 2.94 - 2.92 (m, 2H), 2.92 - 2.87 (m, 2H), 2.86 - 2.83 (m, 2H) 1.74 - 1.66 (m, 1H), 1.56 - 1.48 (m, 1H), 1.11 - 1.08 (m, 4H), 0.99 - 0.92 (m, 2H), 0.89 - 0.87 (m, 5H), 0.82 - 0.73 (m, 2H).EXAMPLE A115. SYNTHESIS OF TWO ATROPISOMERS OF (2S)-N-[(8S,14S,20P)-22-ETHYL-21 -{4-[(1S)-1 -METHOXYETHYL]PYRIDIN-3-IL}-1 8,18-DIMETHYL-9,15-DIOXO-16-OXA-10,22,28-TRIAZAPENTACYCLOri 8.5.2.12,e.11o,14,023,27]NONACOSA-1 (26)

[00321] Step 1. A 1 L round bottom flask was charged with tert-butyl((63S,4S)-12-iodo-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (22.00 g, 32.042 mmol, 1.00 equiv), toluene (300.00 mL), Pd2(dba)3 (3.52 g, 3.845 mmol, 0.12 equiv), S-Phos (3.95 g, 9.613 mmol, 0.30 equiv), and KOAc (9.43 g, 96.127 mmol, 3.00 equiv) at room temperature. To the mixture was added 4,4,5,5-tetramethyl-1,3,2-dioxaborolane (26.66 g, 208.275 mmol, 6.50 equiv) dropwise with stirring at room temperature. The resulting solution was stirred for 3 h at 60 °C. The resulting mixture was filtered and the filter cake was washed with EtOAc. The filtrate was concentrated under reduced pressure, and the remaining residue was purified by silica gel column chromatography to give ((63S,4S)-10,10-dimethyl-5,7-dioxo-12-(4,4,5,5-tetramethyl- 1,3,2-dioxaborolan-2-yl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (22 g, 90%) as a light yellow solid. ESI-MS m/z = 687.3 [M+H]+; Calcd. MW: 686.4

[00322] Step 2. A mixture of tert-butyl ((63S,4S)-10,10-dimethyl-5,7-dioxo-12-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)- 61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine- 2(1,3)-benzenecycloundecaphane-4-yl)carbamate(2.0 g, 2.8 mmol), 3-bromo-2-[(1S)-1-methoxyethyl]pyridine (0.60 g, 2.8 mmol), Pd(dppf)Cl2 (0.39 g, 0.5 mmol), and K3PO4 (1.2 g, 6.0 mmol) in 1,4-dioxane (50 mL) and H2O (10 mL) under a N2 atmosphere was heated to 70 C and stirred for 2 h. The mixture was diluted with H2O (50 mL) and extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with brine (3 × 50 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give tert-butyl((63S,4S)-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (1.5 g, 74% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C40H49N5O6 695.4; found 696.5.

[00323] Step 3. A mixture of tert-butyl ((63S,4S)-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexa- hydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundephane-4-yl)carbamate (1.5 g, 2.1 mmol), Cs2CO3 (2.1 g, 6.3 mmol) and ethyl iodide (0.43 mL, 5.1 mmol) in DMF (50 mL) was stirred at 0 C for 16 h. The mixture was quenched at 0 C with H2O and extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with brine (3 × 50 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give tert-butyl((63S,4S)-11-ethyl- 12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (1.5 g, 99% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C42H53N5O6 723.4; found 724.6.

[00324] Step 4. A mixture of tert-butyl ((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo- 61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundecaphane-4-yl)carbamate (1.3 g, 1.7 mmol) in TFA (10 mL) and DCM (20 mL) was stirred at 0 C for 2 h. The mixture was concentrated under reduced pressure to afford (63S,4S)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-5,7-dione (1.30 g, crude) as a solid. LCMS (ESI): m/z [M+H] calcd for C37H45N5O4 623.3; found 624.4.

[00325] Step 5. In a purged 40 mL flask kept with an inert atmosphere of Ar, (63S,4S)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexa- hydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundephane-5,7-dione (250 mg, 0.4 mmol), (2S)-3-methyl-acid 2-[N-methyl-1-[(3S)-1-(prop-2-enoyl)pyrrolidin-3-yl]formamido]butanoic acid (226mg, 0.8 mmol), DIPEA (774 mg, 6.0 mmol) and DMF (3 mL). A solution of COMU (257 mg, 0.6 mmol) in DMF (2 mL) was added at 0 °C and the resulting mixture was stirred at 0 °C for 1 h. The mixture was filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by preparative HPLC to give two atropisomers of (2S)-N-[(8S,14S,20P)-22-ethyl-21-{4-[(1S)-1-methoxyethyl]pyridin-3-yl}-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6 110.14 023.27]nonacosa-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]-3-methyl-2-{N-methyl-1-[(3S)-1-(prop-2-enoyl)pyrrolidin-3-yl]formamido}butanamide (56 mg, 15% yield) and (46 mg, 13% yield) both as a solid. LCMS (ESI): m/z [M+H] calculated for C51H65N7O7 887.5; found 888.4; 1H NMR (400 MHz, DMSO-d6) δ 8.81 (s, 1H), 8.07 (s,1H), 8.05 - 7.96(m, 1H), 7.78 - 7.45 (m, 5H), 7.41 - 7.08(m, 2H), 6.66 -6.58 (m, 1H), 6.18 (d, J = 17.0 Hz, 1H), 5.75 - 5.67 (m, 1H), 5.46 - 5.31(m, 1H), 5.16 - 5.04 (m, 1H), 4.75 (dd, J = 10.9, 4.5 Hz, 1H), 4.31 - 4.21 (m, 2H), 4.11 - 3.95 (m, 3H), 3.87 - 3.71 (m, 5H), 3.74 - 3.54 (m, 3H), 3.11 (s, 4H), 2.95 (d, J = 9.7 Hz, 2H), 2.85 - 2.72 (m, 3H), 2.31 - 2.04 (m, 3H), 1.88 - 1.47(m, 2H), 1.24 - 1.21 (m, 3H), 1.16 - 1.08 (m, 3H), 1.03 - 0.91 (m, 6H), 0.85 -0.74 (m, 3H), 0.51 - 0.46 (m, 3H) and LCMS (ESI): m/z [M+H] calculated for C51H65N7O7 887.5; found 888.4; 1H NMR (400 MHz, DMSO-d6) δ 8.77 (s, 1H), 8.71 - 8.63 (m, 0.5H), 8.23 - 8.17 (m, 0.5H), 8.00 (s, 1H), 7.85 (t, J = 9.9 Hz, 2H), 7.77 - 7.62 (m, 3H), 7.57 - 7.50 (m, 1H), 7.33 - 7.22 (m, 1H), 7.15 - 7.06 (m, 1H), 6.73 - 6.56 (m, 1H), 6.17 (ddd, J = 16.7, 6.1, 2.7 Hz, 1H), 5.76 - 5.64 (m,, 1H), 5.49 - 5.29 (m, 2H), 4.70 (dd, J = 10.8, 3.5 Hz, 1H), 4.33 - 4.22 (m, 3H), 4.14 - 3.95 (m, 2H), 3.86 - 3.77 (m, 1H), 3.72 - 3.65 (m, 2H), 3.61 (t, J = 10.6 Hz, 3H), 3.46 - 3.42 (m, 1H), 3.13 (d, J = 4.8 Hz, 3H), 2.99 (d, J = 14.4 Hz, 1H), 2.95 - 2.70 (m, 6H), 2.24 - 1.99 (m, 4H), 1.95 - 1.44 (m, 4H), 1.40 (d, J = 6.1 Hz, 3H), 0.98 - 0.87 (m, 6H), 0.86 - 0.64 (m, 6H), 0.64 - 0.54 (m, 3H).EXAMPLE A2: SYNTHESIS OF (2S)-N-[(8S,14S)-4-AMINO-22-ETHYL-21-[2-(2-METHOXYETHYL)PHENYL]-18,18-DIMETHYL-9,15-DIOXO-16-OXA-10,22,28- TRIAZAPENTACYCLE[18.5.2.12,6.110,14.023,27]NONACOSA-1 (26)

[00326] Step 1. In a 25 mL sealed tube, 3-[1-ethyl-2-[2-(methoxymethyl)phenyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indol-3-yl]-2,2-dimethylpropan-1-ol (590 mg, 1.2 mmol) was added. (2S)-3-(3-bromo-5-nitrophenyl)-2-[(tert-butoxycarbonyl)amino]propanoate (747 mg, 1.9 mmol), XPhos Pd G3 (105 mg, 0.12 mmol), XPhos (71 mg, 0.15 mmol), K2CO3 (427 mg, 3.1 mmol) and 1,4-dioxane (2 mL) under an atmosphere of N2 at rt. The mixture was heated to 60 °C and stirred overnight, then cooled and H2O added. The mixture was extracted with EtOAc (3 × 20 mL), and the combined organic layers were washed with brine (1 × 20 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain (2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-(methoxymethyl)phenyl]indol-5-yl]-5-nitrophenyl]propanoic acid (500 mg, 61% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C37H45N3O8 659.3; found 660.4.

[00327] Step 2. A mixture of (2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-(methoxymethyl)phenyl]indol-5-yl]-5-nitrophenyl]propanoic acid (500 mg, 0.79 mmol), methyl (3S)-1,2-diazinane-3-carboxylate (164 mg, 1.1 mmol), DCM (6 mL), DIPEA (294 mg, 2.3 mmol), and HATU (432 mg, 1.1 mmol) was stirred at 0 °C for 1 h under an air atmosphere. H2O was added and the mixture was extracted with DCM (3 × 20 mL), then the combined organic layers were dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain methyl (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-(methoxymethyl)phenyl]indol-5-yl]-5-nitrophenyl]propanoyl]-1,2-diazinane-3-carboxylate (520 mg, 87% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C43H55N5O9 785.4; found 786.8

[00328] Step 3. In a sealed 40 mL tube were added methyl (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-(methoxymethyl)phenyl]indol-5-yl]-5-nitrophenyl]propanoyl]-1,2-diazinane-3-carboxylate (510 mg, 0.65 mmol), DCE (5 mL), and trimethyltin hydroxide (587 mg, 3.3 mmol) at rt under an air atmosphere. The mixture was heated to 60 °C and stirred overnight, cooled, and diluted with DCM (20 mL). The mixture was washed with 0.1 N KHSO4 (3 × 20 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to give ((3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-(methoxymethyl)phenyl]indol-5-yl]-5-nitrophenyl]propanoyl]-1,2-diazinane-3-carboxylic acid (500 mg, 100%) as a solid. LCMS (ESI): m/z [M+H] calcd for C42H53N5O9 771.4; found 772.7.

[00329] Step 4. A mixture of (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-(methoxymethyl)phenyl]indol-5-yl]-5-nitrophenyl]propanoyl]-1,2-diazinane-3-carboxylic acid (490 mg, 0.64 mmol), DCM (100 mL), DIPEA (2.5 g, 19.0 mmol), HOBT (429 mg, 3.2 mmol), and EDCI (3.65 g, 19.0 mmol) at rt was stirred at rt overnight under an atmosphere of air. H2O was added and the mixture was extracted with DCM (3 × 60 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain tert-butyl((63S,4S)-11-ethyl-12-(2-(methoxymethyl)phenyl)-10,10-dimethyl-25-nitro-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (350 mg, 73% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C42H51N5O8 753.4; found 754.2.

[00330] Step 5. A mixture of tert-butyl((63S,4S)-11-ethyl-12-(2-(methoxymethyl)phenyl)-10,10-dimethyl-25-nitro-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (200 mg, 0.27 mmol), MeOH (4 mL), and Pd on carbon (20 mg) was stirred at rt for 2 h under an H2 atmosphere. The mixture was filtered, the filter cake was washed with MeOH (3 × 5 mL), and the filtrate was concentrated under reduced pressure to give tert-butyl((63S,4S)-25-amino-11-ethyl-12-(2-(methoxymethyl)phenyl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (60 mg, 31% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C42H53N5O6 723.4; found 724.4.

[00331] Step 6. In an 8 mL vial were added tert-butyl((63S,4S)-25-amino-11-ethyl-12-(2-(methoxymethyl)phenyl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (50 mg, 0.07 mmol), DCM (1 mL), and TFA (158 mg, 1.4 mmol) at 0 °C under an atmosphere of air. The mixture was stirred at 0 °C for 2 h and then concentrated under reduced pressure to give (63S,4S)-25,4-diamino-11-ethyl-12-(2-(methoxymethyl)phenyl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-5,7-dione (45 mg) as a solid, which was used directly in the next step without further purification. LCMS (ESI): m/z [M+H] calcd for C37H45N5O4 623.3; found 624.4.

[00332] Step 7. Into an 8 mL vial were added (63S,4S)-25,4-diamino-11-ethyl-12-(2-(methoxymethyl)phenyl)-10,10-dimethyl- 61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-5,7-dione (40 mg, 0.06 mmol), DMF (1 mL), DIPEA (75 mg, 0.58 mmol), and COMU (41 mg, 0.1 mmol) at 0 °C under an air atmosphere. The mixture was stirred at 0 °C for 1 h, then H2O was added. The mixture was extracted with EtOAc (3 × 30 mL), the combined organic layers were concentrated under reduced pressure, and purified by preparative HPLC to give (2S)-N-[(8S,14S)-4-amino-22-ethyl-21-[2-(2-methoxyethyl)phenyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6 110.14 023.27]nonacosa- 1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]-3-methyl-2-{N-methyl-1-[(3S)-1-(prop-2-enoyl)pyrrolidin-3-yl]formamido}butanamide (2.5 mg, 4.4% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C51H65N7O7 887.5; found 888.6; 1H NMR (400 MHz, DMSO-d6) δ 8.74 - 8.55 (m, 1H), 7.89 (d, J = 9.6 Hz, 1H), 7.66 - 7.53 (m, 1H), 7.57 - 7.47 (m, 6H), 7.32 (t, J = 6.4 Hz, 1H), 6.85 (d, J = 8.4 Hz, 2H), 6.70 - 6.55 (m, 1H), 6.24 - 6.12 (m, 1H), 5.69 (ddd, J = 14.8, 8.0, 3.9 Hz, 1H), 5.41 (s, 1H), 5.09 - 4.80 (m, 2H), 4.26 (d, J = 10.1 Hz, 2H), 4.19 (s, 2H), 4.17 - 4.06 (m, 1H), 4.02 (dd, J = 12.0, 3.9 Hz, 1H), 3.92 (d, J = 8.0 Hz, 3H), 3.78 (d, J = 8.7 Hz, 5H), 3.29 (s, 2H), 3.14 (d, J = 1.9 Hz, 1H), 2.98 - 2.92 (m, 1H), 2.87 - 2.68 (m, 3H), 2.62 (d, J = 12.5 Hz, 3H), 2.15 - 1.99 (m, 4H), 1.80 (s, 1H), 1.68 - 1.53 (m, 2H), 1.08 (t, J = 7.1 Hz, 1H), 0.98 - 0.88 (m, 6H), 0.82 (dd, J = 23.3, 16.4 Hz, 3H), 0.74 (t, J = 7.2 Hz, 3H), 0.44 (s, 2H), 0.43 (s, 3H).EXAMPLE A118. SUMMARY OF (2S)-N-[(7S,13S)-21-ETHYL-20-[2-(METHOXYMETHYL)PYRIDIN-3-YL]-17,17-DIMETHYL-8,14-DIOXO-15-OXA-3-TIA-9,21,27,28-TETRA-AZAPENTACYCLE[17.5.2.12,5.1 9,13,022,26]OCTACOSA-1(25),2(28),4,19,22(26),23-HEXAEN-7-IL]-3-METHYL-2-{N-METHYL-1-[(3S)-1-(PROP-2-ENOYL)PIRROLIDIN-3-YL]FORMAMIDO}BUTANAMIDE

[00333] Step 1. A mixture of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(1,3-thiazol-4-yl)propanoate (2.08 g, 7.26 mmol) and mCPBA (1.88 g, 10.9 mmol) in DCE (15 mL) at 0 °C under a N2 atmosphere was diluted with DCM (100 mL). The mixture was allowed to warm to rt and stirred for 16 h, then diluted with DCM, washed with H2O (1 x 30 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 4-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-methoxy-3-oxopropyl]-1,3-thiazol-3-io-3-olate (1.15 g, 47% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C12H18N2O5S 302.1; found 303.2.

[00334] Step 2. To a mixture of 4-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-methoxy-3-oxopropyl]-1,3-thiazol-3-io-3-olate (1.15 g, 3.8 mmol) in THF at 0 °C under a N2 atmosphere was added NBS (0.74 g, 4.2 mmol) dropwise. The mixture was allowed to warm to rt and stirred for 2 h, then diluted with H2O (500 mL) and extracted with EtOAc (3 x 500 mL). The combined organic layers were washed with water (2 x 30 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 2-bromo-4-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-methoxy-3-oxopropyl]-1,3-thiazol-3-io-3-olate (1.2 g, 74% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C12H17BrN2O5S 380.0; found 381.0.

[00335] Step 3. To a stirred mixture of 2-bromo-4-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-methoxy-3-oxopropyl]-1,3-thiazol-3-io-3-olate (1.2 g, 3.2 mmol) and 4,4,5,5-tetramethyl-2-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (1.04 g, 4.1 mmol) in MeCN at 70 °C under a N2 atmosphere was added ethane-1,2-diamine (1.89 g, 31.5 mmol) in portions. The mixture was cooled to 60 °C and the mixture was stirred overnight, then diluted with water (500 mL) and extracted with EtOAc (3 x 400 mL). The combined organic layers were washed with brine (1 × 50 mL), dried over anhydrous Na 2 SO 4 , and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give methyl (2S)-3-(2-bromo-1,3-thiazol-4-yl)-2-[(tert-butoxycarbonyl)amino]propanoate (653 mg, 54% yield) as a solid.

[00336] Step 4. A 50 mL sealed tube was charged with 3-[1-ethyl-2-[2-(methoxymethyl)pyridin-3-yl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indol-3-yl]-2,2-dimethylpropan-1-ol (1.00 g, 2.1 mmol), K2CO3 (727 mg, 5.2 mmol), Pd(dppf)Cl2 (153 mg, 0.21 mmol), and 2,4-dibromo-1,3-thiazole (1.0 g, 4.2 mmol) at rt under a N2 atmosphere, then 1,4-dioxane (1.0 mL) and H2O (0.20 mL) were added. The mixture was heated to 70 °C and stirred for 4 h, then cooled, diluted with H2O (100 mL), and extracted with EtOAc (3 × 100 mL). The combined organic layers were washed with brine (3 × 100 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 3-[5-(4-bromo-1,3-thiazol-2-yl)-1-ethyl-2-[2-(methoxymethyl)pyridin-3-yl]indol-3-yl]-2,2-dimethylpropan-1-ol (727 mg, 67% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C34H44N4O6S 636.3; found 637.3.

[00337] Step 5. To a stirred mixture of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-[2-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-(methoxymethyl)pyridin-3-yl]indol-5-yl]-1,3-thiazol-4-yl]propanoate (636 mg, 1.0 mmol) and LiOH.H2O (126 mg, 3.0 mmol) in THF at 0 °C under a N2 atmosphere was added H2O (1.24 mL) in portions. The mixture was allowed to warm to rt and stirred for 1 h, then diluted with water (300 mL) and extracted with EtOAc (3 × 300 mL). The combined organic layers were washed with brine (1 × 100 mL), dried over anhydrous Na2SO4, filtered, and the filtrate concentrated under reduced pressure to give (2S)-2-[(tert-butoxycarbonyl)amino]-3-[2-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-(methoxymethyl)pyridin-3-yl]indol-5-yl]-1,3-thiazol-4-yl]propanoic acid (622 mg, crude), which was used in the next step directly without further purification. LCMS (ESI): m/z [M+H] calcd for C33H42N4O6S 622.3; found 623.2.

[00338] Step 6. To a stirred mixture of (2S)-2-[(tert-butoxycarbonyl)amino]-3-[2-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-(methoxymethyl)pyridin-3-yl]indol-5-yl]-1,3-thiazol-4-yl]propanoic acid (622 mg, 1.0 mmol) and methyl (3S)-1,2-diazinane-3-carboxylate (288 mg, 2.0 mmol) in DMF at 0 °C under a N2 atmosphere was added HATU (570 mg, 1.5 mmol). The mixture was stirred at 0 °C for 1 h, then diluted with EtOAc and washed with H 2 O (1 × 10 mL), dried over anhydrous Na 2 SO 4 , filtered, and the filtrate concentrated under reduced pressure to obtain methyl (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[2-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-(methoxymethyl)pyridin-3-yl]indol-5-yl]-1,3-thiazol-4-yl]propanoyl]-1,2-diazinane-3-carboxylate (550 mg, 62% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C 39 H 52 N 6 O 7 S 748.4; found 749.6.

[00339] Step 7. (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[2-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-(methoxymethyl)pyridin-3-yl]indol-5-yl]-1,3-thiazol-4-yl]propanoyl]-1,2-diazinane-3-carboxylic acid was synthesized in a manner similar to (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl]-5-[(triisopropylsilyl)oxy]phenyl]propanoyl]-1,2-diazinane-3-carboxylic acid except methyl (3S)-1- [(2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl]-5-[(triisopropylsilyl)oxy]phenyl]propanoyl]-1,2-diazinan-3-carboxylate was replaced by methyl (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[2-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-(methoxymethyl)pyridin-3-yl]indol-5-yl]-1,3-thiazol-4-yl]propanoyl]-1,2-diazinane-3-carboxylate. LCMS (ESI): m/z [M+H] calculated for C38H50N6O7S 734.3; found 735.3.

[00340] Step 8. tert-butyl ((63S,4S,Z)-11-ethyl-12-(2-(methoxymethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8- oxa-2(2,4)-thiazole-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-4-yl)carbamate was synthesized in a similar way to tert-butyl ((63S,4S)-11-ethyl-12-(2-((S)-1- methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-25-((triisopropylsilyl)oxy)-61,62,63,64,65,66- hexahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundephane-4-yl)carbamateexcept (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-[(1S)-1- methoxyethyl]pyridin-3-yl]indol-5-yl]-5-[(triisopropylsilyl)oxy]phenyl]propanoyl]-1,2-diazinane-3- carboxylic acid was replaced by (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[2-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-(methoxymethyl)pyridin-3-yl]indol-5-yl]-1,3-thiazol-4-yl]propanoyl]-1,2-diazinane-3-carboxylic acid. LCMS (ESI): m/z [M+H] calcd for C38H48N6O6S 716.3; found 717.4.

[00341] Step 9. To a stirred mixture of tert-butyl ((63S,4S,Z)-11-ethyl-12-(2-(methoxymethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexa- hydro-11H-8-oxa-2(2,4)-thiazole-1(5,3)-indole-6(1,3)-pyridazinecycloundephane-4-yl)carbamate (253 mg) in DCM at 0 C under an N2 atmosphere was added TFA (1.0 mL) dropwise. The mixture was stirred at 0 °C for 1 h, then concentrated under reduced pressure and then repeated using toluene (20 mL × 3) to give (63S,4S,Z)-4-amino-11-ethyl-12-(2-(methoxymethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(2,4)-thiazole-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-5,7-dione (253 mg, crude) as a solid. LCMS (ESI): m/z [M+H] calcd for C33H40N6O4S 616.3; found 617.3.

[00342] Step 10. (2S)-N-[(7S,13S)-21-ethyl-20-[2-(methoxymethyl)pyridin-3-yl]-17,17-dimethyl-8,14-dioxo-15-oxa-3-thia-9,21,27,28-tetra-azapentacyclo[17.5.2.12.5 19,13022,26]octacosa-1(25),2(28),4,19,22(26),23-hexaen-7-yl]-3-methyl-2-{N-methyl-1-[(3S)-1-(prop-2-enoyl)pyrrolidin-3-yl]formamido}butanamide was synthesized in a similar way (2S)-N-[(8S,14S)-4-amino-22-ethyl-21-[2-(2-methoxyethyl)phenyl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28- triazapentaciclo[18.5.2.12,6.i1o,14,023,27]nonacosa-i(26),2,4,6(29),20,23(27),24— heptaen-8-yl]-3-methyl-2-{N-methyl-1-[(3S)-1-(prop-2-enoyl)pyrrolidin-3- yl]formamido}butanamideexcept (63S,4S)-4-amino-11-ethyl-25-hydroxy-12-(2-((S)-1- methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundecaphane-5,7-dione was replaced by (63S,4S,Z)-4-amino-11-ethyl-12-(2-(methoxymethyl)pyridin-3-yl)-10,10-dimethyl- 61,62,63,64,65,66-hexahydro-11H-8-oxa-2(2,4)-thiazole-1(5,3)-indole-6(1,3)- pyridazinecycloundecaphane-5,7-dione. LCMS (ESI): m/z [M+H] calculated for C47H60N8O7S 880.4; found 881.6; 1H NMR (400 MHz, DMSO-d6) δ 8.75 (m, 1H), 8.55 (d, J = 6.7 Hz, 1H), 8.32 (d, J = 8.3 Hz, 1H), 7.99 (d, J = 7.7 Hz, 1H), 7.65 - 7.51 (m, 3H), 7.11 - 6.92 (m, 1H), 6.72 - 6.56 (m, 1H), 6.18 (dd, J = 16.8, 2.9 Hz, 1H), 5.82 - 5.65 (m, 1H), 5.61 - 5.46 (m, 1H), 5.02 (dd, J = 24.2, 12.2Hz, 1H), 4.69 (d, J = 10.9 Hz, 1H), 4.37 - 4.11 (m, 5H), 4.05 - 3.79 (m, 4H), 3.76 - 3.50 (m, 6H), 3.47 (s, 2H), 3.08 (s, 3H), 3.04 (s, 1H), 2.98 (d, J = 1.9 Hz, 1H), 2.95 (d, J = 3.6 Hz, 2H), 2.83 (d, J = 2.0 Hz, 2H), 2.24 - 2.03 (m, 4H), 1.81 (s, 2H), 1.56 (s, 1H), 1.11 (t, J = 7.0 Hz, 2H), 1.02 - 0.87 (m, 8H), 0.80 (dd, J = 24.6, 6.6 Hz, 3H), 0.41 (s, 2H), 0.31 (s, 1H).EXAMPLE A194. SYNTHESIS OF (2S)-N-[(7S,13S)-21-ETHYL-20-[2-(METHOXYMETHYL)PYRIDIN-3-YL]-17,17-DIMETHYL-8,14-DIOXO-15-OXA-4-TIA-9,21,27,28- TETRA-AZAPENTACYCLO[17.5.2.12,5.19,13.022,26]OCTACOSA- 1(25),2.5(28),19,22(26),23-HEXAEN-7-IL]-3-METHYL-2-{N-METHYL-1-[(3S)-1-(PROP-2- ENOIL)PYRROLIDIN-3-YL]FORMAMIDO}BUTANAMIDE

[00343] Step 1. A mixture of Zn (1.2 g, 182 mmol) and 1,2-dibromoethane (1.71 g, 9.1 mmol) and DMF (50 mL) was stirred for 30 min at 90 °C under an Ar atmosphere. The mixture was allowed to stand at rt, then TMSCl (198 mg, 1.8 mmol) was added dropwise over 30 min at rt. Methyl (2R)-2-[(tert-butoxycarbonyl)amino]-3-iodopropanoate (10.0 g, 30.4 mmol) in DMF (100 mL) was added dropwise over 10 min at rt. The mixture was heated to 35 °C and stirred for 2 h, then a mixture of 2,5-dibromo-1,3-thiazole (1.48 g, 60.8 mmol) and Pd(PPh3)2Cl2 (2.1 g, 3.0 mmol) in DMF (100 mL) was added dropwise. The mixture was heated to 70 °C and stirred for 2 h, then filtered and the filtrate diluted with EtOAc (1 L) and washed with H2O (3 × 1 L), dried with anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give methyl (2S)-3-(5-bromo-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)amino]propanoate (3 g, 27% yield) as a semisolid. LCMS (ESI): m/z [M+H] calcd for C12H17BrN2O4S 364.0; found 365.1.

[00344] Step 2. In a sealed 20 mL tube, added 3-[1-ethyl-2-[2-(methoxymethyl)pyridin-3-yl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indol-3-yl]-2,2-dimethylpropan-1-ol (100 mg, 0.21 mmol), K3PO4 (111 mg, 0.52 mmol), Pd(dppf)Cl2 (15 mg, 0.02 mmol), methyl (2S)-3-(4-bromo-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)amino]propanoate (153 mg, 0.42 mmol), toluene (1 mL), and H2O (0.2 mL) at rt under N2 atmosphere. The mixture was heated to 60 °C and stirred for 3 h, cooled, diluted with H2O (10 mL), and extracted with EtOAc (10 mL × 3). The combined organic layers were washed with brine (3 × 10 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-[4-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-(methoxymethyl)pyridin-3-yl]indol-5-yl]-1,3-thiazol-2-yl]propanoate (72 mg, 54% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C34H44N4O6S 636.3; found 637.2.

[00345] Step 3. A mixture of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-[4-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-(methoxymethyl)pyridin-3-yl]indol-5-yl]-1,3-thiazol-2-yl]propanoate (40 mg, 0.06 mmol) and LiOH.H2O (unspecified) in THF (1 mL) and H2O (0.2 mL) was stirred at rt under a N2 atmosphere for 2 h. The mixture was acidified to pH 5 with aqueous NaHSO4 and extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine (3 × 10 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give (2S)-2-((tert-butoxycarbonyl)amino)-3-(4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-(methoxymethyl)pyridin-3-yl)-1H-indol-5-yl)thiazol-2-yl)propanoic acid. The crude product was used in the next step directly without further purification. LCMS (ESI): m/z [M+H] calcd for C33H42N4O6S 622.3; found 623.3.

[00346] Step 4. Methyl (3S)-1-((2S)-2-((tert-butoxycarbonyl)amino)-3-(4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2- (methoxymethyl)pyridin-3-yl)-1H-indol-5-yl)thiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylate was synthesized in a similar manner to methyl (3S)-1-[(2S)-2- [(tert-butoxycarbonyl)amino]-3-[2-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-(methoxymethyl)pyridin-3-yl]indol-5-yl]-1,3-thiazol-4-yl]propanoyl]-1,2-diazinan-3- carboxylate, except (2S)-2-[(tert-butoxycarbonyl)amino]-3-[2-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-(methoxymethyl)pyridin-3-yl]indol-5-yl]-1,3-thiazol-4-yl]propanoic acid was replaced by (2S)-2-((tert-butoxycarbonyl)amino)-3-(4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-(methoxymethyl)pyridin-3-yl)-1H-indol-5-yl)thiazol-2-yl)propanoic acid. LCMS (ESI): m/z [M+H] calculated for C39H52N6O7S 748.4; found 749.4.

[00347] Step 5. (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[4-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-(methoxymethyl)pyridin-3-yl]indol-5-yl]-1,3-thiazol-2-yl]propanoyl]-1,2-diazinane-3-carboxylic acid was synthesized in a manner similar to (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl]-5-[(triisopropylsilyl)oxy]phenyl]propanoyl]-1,2-diazinane-3-carboxylic acid except methyl methyl (3S)-1-((2S)-2-((tert-butoxycarbonyl)amino)-3- (4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-(methoxymethyl)pyridin-3-yl)-1H-indol-5-yl)thiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylate. LCMS (ESI): m/z [M+H] calculated for C38H50N6O7S 734.3; found 735.4.

[00348] Step 6. Tert-butyl ((63S,4S,Z)-11-ethyl-12-(2-(methoxymethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H- 8-oxa-2(4,2)-thiazole-1(5,3)-indole-6(1,3)-pyridazinecycloundephane-4-yl)carbamate was synthesized in a similar manner to tert-butyl ((63S,4S)-11- ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-25-((tri- (isopropylsilyl)oxy)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate except (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-5-yl]-5-[(triisopropylsilyl)oxy]phenyl]propanoyl]-1,2-diazinane-3-carboxylic acid was replaced by (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[4-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-(methoxymethyl)pyridin-3-yl]indol-5-yl]-1,3-thiazol-2-yl]propanoyl]-1,2-diazinane-3-carboxylic. LCMS (ESI): m/z [M+H] calculated for C38H48N6O6S 716.3; found 717.3.

[00349] Step 7. (63S,4S,Z)-4-amino-11-ethyl-12-(2-(methoxymethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa- 2(4,2)-thiazole-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-5,7-dione was synthesized in a similar way to (63S,4S,Z)-4-amino-11-ethyl-12-(2-(methoxymethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa- 2(2,4)-thiazole-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-5,7-dione except tert-butyl ((63S,4S,Z)-11-ethyl-12-(2-(methoxymethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(2,4)-thiazole-1(5,3)-indole-6(1,3)- pyridazinecycloundecaphane-4-yl)carbamate was replaced by tert-butyl((63S,4S,Z)-11-ethyl-12-(2-(methoxymethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo- 61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-thiazole-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-4-yl)carbamate. LCMS (ESI): m/z [M+Na] calcd for C33H40N6O4SNa 639.3; found 640.3.

[00350] Step 8. (2S)-N-[(7S,13S)-21-ethyl-20-[2-(methoxymethyl)pyridin-3-yl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetra- azapentacyclo[17.5.2.12,5.19,13.022,26]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-3-methyl-2-{N-methyl-1-[(3S)-1-(prop-2-enoyl)pyrrolidin-3-yl]formamido}butanamide was synthesized in a similar way to (2S)-N-[(7S,13S)- 21-ethyl-20-[2-(methoxymethyl)pyridin-3-yl]-17,17-dimethyl-8,14-dioxo-15-oxa-3-thia- 9,21,27,28-tetra-azapentacyclo[17.5.2.12,5.i9,i3.022.26]octacosa- 1(25),2(28),4,19,22(26),23-hexaen-7-yl]-3-methyl-2-{N-methyl-1-[(3S)-1-(prop-2-enoyl)pyrrolidin-3-yl]formamido}butanamide except (63S,4S)-4-amino-11-ethyl-25-hydroxy-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexa- hydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundephane- 5,7-dione was replaced by (63S,4S,Z)-4-amino-11-ethyl-12-(2-(methoxymethyl)pyridin- 3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-thiazola-1(5,3)- indole-6(1,3)-pyridazinecycloundecaphane-5,7-dione. LCMS (ESI): m/z [M+H] calculated for C47H60N8O7S 880.4; found 881.5; 1H NMR (400 MHz, DMSO-d6) δ 8.70 (dt,J= 16.2, 8.1 Hz, 1H), 8.54 (ddd, J = 6.6, 4.7, 1.7 Hz, 1H), 8.50 (m, 1H), 7.96 (d, J = 7.8 Hz, 1H), 7.88 (t, J = 2.1 Hz, 2H), 6.70 - 6.57 (m, 2H), 6.24 - 6.13 (m, 2H), 5.75 (m, 1H), 5.55 (t, J = 7.3 Hz, 1H), 5.46 (d, J = 8.5 Hz, 1H), 5.14 (d, J = 13.0 Hz, 1H), 4.84 - 4.75 (m, 1H), 4.35 (d, J = 10.7 Hz, 1H), 4.28 - 4.19 (m, 4H), 3.91 (s, 3H), 3.87 (dd, J = 10.4, 8.1 Hz, 1H), 3.78 - 3.70 (m, 2H), 3.63 (t, J = 8.8 Hz, 2H), 3.61 - 3.49 (m, 2H), 2.87 (d, J = 1.1 Hz, 2H), 2.79 (s, 1H), 2.38 (s, 1H), 2.18 (s, 1H), 2.13 (d, J = 10.7 Hz, 4H), 1.96 (s, 2H) 3H).EXAMPLE A71. SYNTHESIS OF (2S)-2-(1-{1-[(2E)-4-(DIMETHYLAMINO)BUT-2- ENOYL]AZETIDIN-3-IL}-N-METHYLFORMAMIDO)-N-[(8S,14S)-22-ETHYL-4-HYDROXY-21-[2- (METHOXYMETHYL)PYRIDIN-3-YL]-1 8,18-DIMETHYL-9,15-DIOXO-16-OXA-1 0,22,28- TRIAZAPENTACYCLE[18.5.2.12,6.110,14.023,27]NONACOSA- 1(26),2,4,6(29),20,23(27),24-HEPTAEN-8-IL]-3-METHYLBUTANAMIDE

[00351] Step 1. To a mixture of tert-butyl N-(azetidine-3-carbonyl)-N-methyl-L-valinate (350 mg, 1.3 mmol) and (2E)-4-(dimethylamino)but-2-enoic acid (201 mg, 1.56 mmol) in DCM (8 mL) at 5 °C was added a solution of T3P, 50% in EtOAc (827 mg, 2.6 mmol) and DIPEA (1.7 g, 13 mmol) in DCM (2 mL). The mixture was stirred for 1 h, then diluted with EtOAc (20 mL) and H2O (20 mL). The aqueous and organic layers were separated, and the organic layer was washed with H2O (3 × 10 mL), brine (10 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative HPLC to obtain tert-butyl (E)-N-(1-(4-(dimethylamino)but-2-enoyl)azetidine-3-carbonyl)-N-methyl-L-valinate (200 mg, 39% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C20H35N3O4 381.3; found 382.3.

[00352] Step 2. To a mixture of tert-butyl (E)-N-(1-(4-(dimethylamino)but-2-enoyl)azetidine-3-carbonyl)-N-methyl-L-valinate (190 mg, 0.32 mmol) in DCM (3 mL) at rt was added TFA (1 mL). The mixture was stirred at rt for 1 h, then concentrated under reduced pressure to give (E)-N-(1-(4-(dimethylamino)but-2-enoyl)azetidine-3-carbonyl)-N-methyl-L-valine (190 mg, 90%) as a solid, which was used directly in the next step without further purification. LCMS (ESI): m/z [M+H] calcd for C16H27N3O4 325.2; found 326.2.

[00353] Step 3. To a mixture of (63S,4S)-4-amino-11-ethyl-25-hydroxy-12-(2-(methoxymethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexa- hydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundephane-5,7-dione (172 mg, 0.27 mmol) and (E)-N-(1-(4-(dimethylamino)but-2-enoyl)azetidine-3-carbonyl)-N-methyl-L-valine (105 mg, 0.32 mmol) in DMF (2 mL) at 5°C was added a mixture of HATU (133 mg, 0.297 mmol) and DIPEA (348 mg, 2.7 mmol) in DMF (1 mL). The mixture was stirred for 1 h, then diluted with EtOAc (20 mL) and H2O (20 mL). The aqueous and organic layers were separated, and the organic layer was washed with H2O (3 × 10 mL), brine (10 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative TLC to obtain (2S)-2-(1-{1-[(2E)-4-(dimethylamino)but-2-enoyl]azetidin-3-yl}-N-methylformamido)-N-[(8S,14S)-22-ethyl-4-hydroxy-21-[2-(methoxymethyl)pyridin-3-yl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6.110,14.023,27]nonacosa-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]-3-methylbutanamide (4.8 mg, 2% yield over 2 steps) as a solid. LCMS (ESI): m/z [M+H] calculated for C52H68N8O8 932.5; found 933.5; 1H NMR (400 MHz, CD3OD) δ 8.71 (d, J = 3.2 Hz, 1H), 8.50 (s, 1.5H), 8.08 - 7.85 (m, 2H), 7.65 - 7.44 (m, 3H), 7.32 - 7.14 (m, 1H), 7.07 - 6.95 (m, 1H), 6.80 (dt, J = 22.1, 6.8 Hz, 1H), 6.55 (d, J = 35.8 Hz, 1H), 6.30 (d, J = 15.4 Hz, 1H), 5.56 (dd, J = 13.8, 6.7 Hz, 1H), 4.76 (dd, J = 19.8, 10.5 Hz, 1H), 4.54 (dd, J = 15.9, 7.5 Hz, 2H), 4.48 - 4.38 (m, 2H), 4.36 - 4.23 (m, 3H), 4.22 - 4.14 (m, 1H), 3.96 (qd, J = 15.6, 7.9 Hz, 3H), 3.77 (ddd, J = 25.8, 23.4, 11.9 Hz, 2H), 3.58 (dd, J = 17.2, 8.3 Hz, 2H), 3.38 (s, 2H), 3.25 - 3.11 (m, 3H), 3.05 - 2.94 (m, 1H), 2.94 - 2.81 (m, 4H), 2.73 (dd, J = 20.9, 11.0 Hz, 1H), 2.45 (d, J = 6.9 Hz, 5H), 2.32 - 2.07 (m, 3H), 1.92 (d, J = 13.2 Hz, 1H), 1.72 (s, 1H), 1.64 - 1.51 (m, 1H), 1.18 (t, J = 7.0 Hz, 2H), 1.00 (ddd, J = 14.6, 11.8, 8.5 Hz, 6H), 0.92 - 0.81 (m, 4H), 0.55 - 0.41 (m, 3H).EXAMPLE A67. SYNTHESIS OF (2E)-4-(DIMETHYLAMINO)-N-(6-{[(1S)-1-{[(8S,14S)-22- ETHYL-4-HYDROXY-21-[2-(METHOXYMETHYL)PYRIDIN-3-YL]-18,18-DIMETHYL-9,15-DIOXO-16-OXA-10,22,28-TRIAZAPEN TACICLOri8.5.2.12,e.11o,14,023,27]NONACOSA-1(26),2,4,6(29),20,23(27),24-HEPTAEN-8-YL]CARBAMOIL}-2- METHYLPROPYL](METHYL)CARBAMOYL}PYRIDIN-3-YL)BUT-2-ENAMIDE

[00354] Step 1. To a TFA salt mixture of (63S,4S)-4-amino-11-ethyl-25-hydroxy-12-(2-(methoxymethyl)pyridin-3-yl)-10,10-dimethyl- 61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundecaphane-5,7-dione (225 mg, 0.28 mmol) and (E)-N- TFA salt (5-(4-(dimethylamino)but-2-enamido)picolinoyl)-N-methyl-L-valine (260 mg crude, 0.56 mmol) in DMF (5 mL) at 0°C was added DIPEA (0.46 mL, 2.8 mmol) followed by HATU (140 mg, 0.36 mmol). The mixture was stirred at 0-10 °C for 1 h, then concentrated under reduced pressure, and the residue was purified by preparative HPLC to give (2E)-4-(dimethylamino)-N-(6-{[(1S)-1-{[(8S,14S)-22-ethyl-4-hydroxy-21-[2-(methoxymethyl)pyridin-3-yl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6 110.14 023.27]nonacosa- 1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamoyl}-2- methylpropyl](methyl)carbamoyl}pyridin-3-yl)but-2-enamide (23.3 mg, 8% yield over 2 steps) as a solid. LCMS (ESI): m/z [M+Na] calculated for C54H67N9O8Na 992.5; found 992.4; - 7.70 (m, 1H), 7.60 - 7.47 (m, 2H), 7.31 - 7.19 (m, 1H),7.07 - 6.90 (m, 2H), 6.70 - 6.36 (m, 3H), 5.81 - 5.61 (m, 1H), 4.50 - 4.20 (m, 4H),4.01 - 3.68 (m, 3H), 3.64 - 3.35 (m, 5H), 3.27 - 3.08 (m, 3H), 3.04 - 2.44 (m, 11H),2.36 - 2.10 (m, 3H), 1.93 (d, J = 13.0 Hz, 1H), 1.61 (dd, J = 34.3,21.6Hz, 3H), 1.39 - 1.16 (m, 3H), 1.12 - 0.81 (m, 6H), 0.78 -0.45 (m, 6H).EXAMPLE A54. SYNTHESIS OF (2S)-2-{1-[(3S)-1 -[(2E)-4-(DIMETHYLAMINO)BUT-2- ENOYL]PYRROLIDIN-3-YL]-N-METHYLFORMAMIDO}-N-[(8S,14S)-22-ETHYL-4-HYDROXY-21-[2- (METHOXYMETHYL)PYRIDIN-3-IL]-1 8,18-DIMETHYL-9,15-DIOXO-16-OXA-1 0,22,28- TRIAZAPENTACYCLE[18.5.2.12,6.110,14.023,27]NONACOSA-1(26),2,4,6(29),20,23(27),24-HEPTAEN-8-IL]-3-METHYLBUTANAMIDE

[00355] Step 1. To a mixture of tert-butyl N-methyl-N-((S)-pyrrolidine-3-carbonyl)-L-valinate (210 mg, 0.73 mmol) in DMF (4 mL) at rt were added 4-(dimethylamino)-4-methylpent-2-ynoic acid (450 mg, 2.9 mmol), DIPEA (1.2 mL, 7.3 mmol), and HATU (332 mg, 0.88 mmol). The mixture was stirred at rt for 1 h, then diluted with EtOAc and the mixture was washed with H2O, brine, dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give tert-butyl N-((S)-1-(4-(dimethylamino)-4-methylpent-2-ynoyl)pyrrolidine-3-carbonyl)-N-methyl-L-valinate (140 mg, 45% yield) as an oil. LCMS (ESI): m/z [M+H] calcd for C23H39N3O4 421.3; found 422.3.

[00356] Step 2. A mixture of tert-butyl N-((S)-1-(4-(dimethylamino)-4-methylpent-2-ynoyl)pyrrolidine-3-carbonyl)-N-methyl-L-valinate (130 mg, 0.31 mmol) in DCM (2 mL) and TFA (1 mL) was stirred at rt for 90 min. The mixture was concentrated under reduced pressure to give N-((S)-1-(4-(dimethylamino)-4-methylpent-2-ynoyl)pyrrolidine-3-carbonyl)-N-methyl-L-valine TFA salt (150 mg) as an oil, which was used directly in the next step without further purification. LCMS (ESI): m/z [M+H] calcd for C19H31N3O4 365.2; found 366.2.

[00357] Step 3. TFA salt (3S)-1-(4-(dimethylamino)-4-methylpent-2-ynoyl)-N-((2S)-1-(((63S,4S)-11-ethyl-25-hydroxy-12-(2-(methoxymethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro- 11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazin-2(1,3)-benzenacycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)-N-methylpyrrolidine-3-carboxamide was synthesized in a similar manner to 1-acryloyl-N-((2S)-1-(((63S,4S)-11-ethyl-25-hydroxy-12-(2-(methoxymethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66- hexahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundephane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)-N-methylazetidine-3-carboxamide, except TFA salt of (2S)-2-{1-[(3S)-1-[(2E)-4-(dimethylamino)but-2-enoyl]pyrrolidin-3-yl]-N-methylformamido}-N-[(8S,14S)-22-ethyl- 4-hydroxy-21-[2-(methoxymethyl)pyridin-3-yl]-18,18-dimethyl-9,15-dioxo-16-oxa- 10,22,28-triazapentacyclo[18.5.2.12.6 110.14 023.27]nonacosa- 1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]-3-methylbutanamide. (120 mg, 54% yield in 2 steps) as a solid. 1H-NMR (400 MHz, CD3OD) δ 8.76 - 8.68 (m, 1H), 8.44 (s, 1H), 8.02 - 7.94 (m, 1H), 7.94 - 7.84 (m, 1H), 7.65 - 7.43(m, 3H), 7.27 - 7.14 (m, 1H), 7.06 - 6.96 (m, 1H), 6.65 - 6.48 (m, 1H), 5.62 - 5.46(m, 1H), 4.81 - 4.57 (m, 1H), 4.46 - 4.22 (m, 3H), 4.10 - 3.35 (m, 11H), 3.26 - 2.93(m, 6H), 2.91 - 2.51 (m, 4H), 2.42 - 2.09 (m, 9H), 1.95 - 1.87 (m, 1H), 1.85 - 1.40 (m, 6H), 1.38 - 1.10 (m, 6H), 1.07 - 0.81 (m, 9H), 0.56 - 0.38 (m, 3H). LCMS (ESI): m/z [M+H] C52H68N8O8 found 947.7. EXAMPLE A95. SYNTHESIS OF (2S)-N-[(8S,14S)-22-ETHYL-4-HYDROXY-21-[2-(METHOXYMETHYL)PYRIDIN-3-YL]-1 8,18-DIMETHYL-9,15-DIOXO-16-OXA-1 0,22,28-TRIAZAPENTACYCLE[18,5,2,12,6,110,14,023,27]NONACOSA-1 (26),2,4,6(29),20,23(27),24-HEPTAEN-8-IL]-3-METHYL-2-{N-METHYL-1 -[(3S)-1 -[4- (MORFOUN-4-IL)BUT-2-INOIL|PYRROUDIN-3-IL|FORMAMIDO}BUTANAMIDE

[00358] Step 1. A mixture of tert-butyl (2S)-3-methyl-2-[N-methyl-1-(3S)-pyrrolidin-3-ylformamido]butanoate (500 mg, 1.8 mmol), 4-(morpholin-4-yl)but-2-ynoic acid (1.49 g, 8.8 mmol), DIPEA (682 mg, 5.3 mmol), and CIP (635 mg, 2.3 mmol) in DMF (5 mL) was stirred at 0 °C for 2 h. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain tert-butyl N-methyl-N-((S)-1-(4-morpholinobut-2-ynoyl)pyrrolidine-3-carbonyl)-L-valinate (150 mg, 19% yield) as an oil. LCMS (ESI): m/z [M+H] calculated for C23H37N3O5 435.3; found 436.5.

[00359] Step 2. A mixture of tert-butyl N-methyl-N-((S)-1-(4-morpholinobut-2-inoyl)pyrrolidine-3-carbonyl)-L-valinate (250 mg, 0.57 mmol) in DCM (5 mL) and TFA (2.5 mL) was stirred at rt for 2 h. The mixture was concentrated under reduced pressure to give (2S)-3-methyl-2-[N-methyl-1-[(3S)-1-[4-(morpholin-4-yl)but-2-inoyl]pyrrolidin-3-yl]formamido]butanoic acid (310 mg, crude) as an oil, which was used directly in the next step without further purification. LCMS (ESI): m/z [M+H] calcd for C19H29N3O5 379.2; found 380.2.

[00360] Step 3. A mixture of (63S,4S)-4-amino-11-ethyl-25-hydroxy-12-(2-(methoxymethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro- 11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundephane-5,7-dione (250 mg, 0.4 mmol), DIPEA (516 mg, 4.0 mmol), acid (2S)-3-methyl-2-[N- methyl-1-[(3S)-1-[4-(morpholin-4-yl)but-2-inoyl]pyrrolidin-3-yl]formamido]butanoic acid (182 mg, 0.48 mmol) and COMU (205 mg, 0.48 mmol) in DMF (3 mL) was stirred at -20 C for 2 h. The mixture was diluted with H2O (10 mL), then extracted with EtOAc (3 × 10 mL) and the combined organic layers were washed with brine (3 × 10 mL), dried over anhydrous Na2SO4, and filtered. The mixture was concentrated under reduced pressure, and the residue was purified by reverse-phase silica gel column chromatography to give (2S)-N-[(8S,14S)-22-ethyl-4-hydroxy-21-[2-(methoxymethyl)pyridin-3-yl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.12,6 110.14 023.27]nonacosa-1(26),2A6(29),20,23(27),24-heptaen-8-yl]-3-methyl-2-{N-methyl-1-[(3S)-1-[4-(morpholin-4-yl)but-2-inoyl]pyrrolidin-3-yl]formamido}butanamide (207 mg, 53% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C55H70N8O9 986.5; found 987.8; 1H NMR (400 MHz, DMSO-d6) δ 9.39 - 9.28 (m, 1H), 8.74 (t,J = 4.8, 1H), 8.70 - 8.04 (m, 1H), 7.98 - 7.90 (m, 1.5H), 7.82 (d,J = 7.7 Hz, 0.5H), 7.63 - 7.46 (m, 3H), 7.26 - 7.10 (m, 1H), 7.03 (s, 1H), 6.58 - 6.43 (m, 1H), 5.44 - 5.30 (m, 1H), 5.06 (q, 0.5H), 4.72 (t ,J = 11.0, 0.5H), 4.39 - 4.20 (m, 3H), 4.15 (d ,J = 11.1 Hz, 1H), 4.09 - 3.85 (m, 4H), 3.66 (s, 2H), 3.65 - 3.58 (m, 4H), 3.58 - 3.55 (m, 2H), 3.55 - 3.48 (m, 3H), 3.47 - 3.41 (m, 3H), 3.31 (s, 2H), 3.10 (s, 2H), 2.92 (s, 1H), 2.89 - 2.65 (m, 5H), 2.68 (s, 1H), 2.45 - 2.38 (m, 1H), 2.29 - 2.24 (m, 1H), 2.23 - 1.99 (m, 3H), 1.82 (d ,J = 12.1 Hz, 1H), 1.76 - 1.62 (m, 1H), 1.61 - 1.45 (m, 1H), 1.14 - 1.04 (m, 2H), 1.02 - 0.92 (m, 3H), 0.91 - 0.86 (m, 3H), 0.83 - 0.77 (m, 3H), 0.77 - 0.70 (m, 2H), 0.50 - 0.35 (m, 3H).EXAMPLE A145. SYNTHESIS OF TWO ATROPISOMERS (2S)-2-{1-[(3S)-1-(BUT-2-INOIL)PYRROLIDIN-3-IL]-N-METHYLFORMAMIDO}-N-[(8S, 14S,20M)-22-ETHYL-4-HYDROXY-21-{2-[(1S)-1-METHOXYETHYL]PYRIDIN-3-YL}-18,18-D IMETHYL-9,15-DIOXO-16-OXA-10,22,28-TRIAZAPENTACYCLE[18.5.2.1²,⁶.1¹⁰,¹⁴.0²³ ,²⁷]NONACOSA-1(26),2,4,6(29),20,23(27),24-HEPTAEN-8-IL]-3-METHYLBUTANAMIDE

[00361] Step 1. To a mixture of but-2-ynoic acid (222 mg) and CIP (588 mg) in ACN (8 mL) at 0 °C under an Ar atmosphere was added DIPEA (681 mg). The mixture was stirred at 0 °C, then tert-butyl N-methyl-N-((S)-pyrrolidine-3-carbonyl)-L-valinate (500 mg) in ACN (3 mL) was added dropwise and the mixture was stirred at 0 °C for 2 h. EtOAc was added and the mixture was washed with brine (3 x 20 mL), dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give tert-butyl N-((S)-1-(but-2-ynoyl)pyrrolidine-3-carbonyl)-N-methyl-L-valinate as a solid. LCMS (ESI): m/z [M+H] calculated for C19H30N2O4 350.2; found 352.1.

[00362] Step 2. A mixture of tert-butyl N-((S)-1-(but-2-ynoyl)pyrrolidine-3-carbonyl)-N-methyl-L-valinate (200 mg) in DCM (4 mL) and TFA (2 mL) was stirred at 0 °C for 2 h. The mixture was concentrated under reduced pressure with azeotropic removal of H2O using toluene (4 mL x 2) to give N-((S)-1-(but-2-ynoyl)pyrrolidine-3-carbonyl)-N-methyl-L-valinate as a solid. LCMS (ESI): m/z [M+H] calcd for C15H22N2O4 294.2; found 295.2.

[00363] Step 3. Two atropisomers of (2S)-2-{1-[(3S)-1- (but-2-inoyl)pyrrolidin-3-yl]-N-methylformamido}-N-[(8S,14S,20M)-22-ethyl-4-hydroxy- 21-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacycle [18.5.2.1²,⁶.1¹⁰,¹⁴.0²³,²⁷]nonacosa-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]-3-methylbutanamide was synthesized in a similar way to (2S)-N- [(8S,14S)-22-ethyl-4-hydroxy-21-[2-(methoxymethyl)pyridin-3-yl]-18,18-dimethyl-9,15- dioxo-16-oxa-10,22,28-triazapentaciclo[18.5.2.1²,⁶.1¹⁰,¹⁴.0²³,²⁷]nonacosa- 1(26) 3-Methyl-2-[N-methyl-1-[(3S)-1-[4-(morpholin-4-yl)but-2-inoyl]pyrrolidin-3-yl]formamido]butanoic acid was replaced by N-((S)-1-(but-2-inoyl)pyrrolidin-3-carbonyl)-N-methyl-L-valinate. (43.3 mg, 12% yield) and (33 mg, 9% yield) both as solids. LCMS (ESI): m/z [M+H] calculated for C52H65N7O8 915.5; found 916.7; 1H NMR (400 MHz, DMSO-d6) δ 9.34 - 9.27 (m, 1H), 8.78 (t, J = 2.5 Hz, 1H), 8.68 (t, J = 8.5 Hz, 0.5H), 8.20 - 8.11 (m, 0.6H), 7.95 (ddt, J = 5.4, 3.5, 1.7 Hz, 2H), 7.63 - 7.60 (m, 1H), 7.61 - 7.49 (m, 2H), 7.13 (s, 1H), 7.03 (d, J = 6.2 Hz, 1H), 6.60 - 6.49 (d, J = 35.5 Hz, 1H), 5.43 - 5.39 (m, 1H), 5.12 - 5.00 (m, 0.7H), 4.74 (d, J = 10.6 Hz, 0.4H), 4.32 - 4.25 (m, 1H), 4.18 - 3.85 (m, 5H), 3.81 - 3.45 (m, 8H), 3.18 - 3.02 (m, 5H) 1.74 - 1.63 (m, 1H), 1.6 2-1.42 (m, 1H), 1.32 - 1.16 (m, 4H), 1.15 - 1.05 (t, J = 6.3 Hz, 4H), 1.04 - 0.95 (m, 2H), 0.95 - 0.85 (m, 5H), 0.68 - 0.52 (m, 4H), 0.52 - 0.37 (m, 4H). and LCMS (ESI): m/z [M+H] calculated for C52H65N7O8 915.5; found 916.7; 1H NMR (400 MHz, DMSO-d6) δ 9.36 - 9.28 (m, 1H), 8.77 (dd, J = 4.7, 1.8 Hz, 1H), 8.62 - 8.57 (m, 0.5H), 8.15 - 8.07 (m, 0.5H), 7.95 (s, 1H), 7.87 - 7.81 (m, 1H), 7.65 - 7.51 (m, 3H), 7.37 - 7.25 (m, 1H), 7.10 - 7.03 (m, 1H), 6.54 (d, J = 35.5Hz, 1H), 5.52 - 5.21 (m, 2H), 4.78 - 4.66 (m, 0.5H) 2.28 - 1.96 (m, 7H), 1.95 - 1.74 (m, 2H), 1.73 - 1.44 (m, 2H), 1.42 - 1.37 (m, 3H), 1.28 - 1.14 (m, 1H), 1.03 - 0.85 (m, 6H), 0.83 - 0.72 (m, 7H), 0.71 - 0.55 (m, 3H). EXAMPLE A28. SYNTHESIS OF (2S)-N-[(8S,14S)-22-ETHYL-4-HYDROXY-21-[4- (METHOXYMETHYL)PYRIDIN-3-YL]-18,18-DIMETHYL-9,15-DIOXO-16-OXA-10,22,28- TRIAZAPENTACYCLO[18.5.2.1^[2,6].1^[10,14].0^[23,27]]NONACOSA- 1(26),2,4,6(29),20,23(27),24-HEPTAEN-8-IL]-3-METHYL-2-[N-METHYL-1-[(3S)-1-(PROP- 2-ENOIL)PYRROLIDIN-3-YL]FORMAMIDO]BUTANAMIDE

[00364] Step 1. To a mixture of 3-bromo-4-(methoxymethyl)pyridine (1.00 g, 5.0 mmol), 4,4,5,5-tetramethyl-2-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (1.51 g, 5.9 mmol) and KOAc (1.21 g, 12.3 mmol) in toluene (10 mL) at rt under an Ar atmosphere was added Pd(dppf)Cl2 (362 mg, 0.5 mmol). The mixture was heated to 110 °C and stirred overnight, then concentrated under reduced pressure to give 4-(methoxymethyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine, which was used directly in the next step without further purification. LCMS (ESI): m/z [M+H] calcd for C13H20BNO3 249.2; found 250.3.

[00365] Step 2. To a mixture of 4-(methoxymethyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (290 mg, 1.16 mmol), K3PO4 (371 mg, 1.75 mmol) and tert-butyl N-[(8S,14S)-21-iodo-18,18-dimethyl-9,15-dioxo-4-[(triisopropylsilyl)oxy]-16-oxa-10,22,28-triazapentacyclo[18.5.2.1 A[2,6]. 1A[10,14].0A[23,27]]nonacosa-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamate (500 mg, 0.58 mmol) in 1,4-dioxane (5 mL) and H2O (1 mL) at rt under an Ar atmosphere was added Pd(dppf)Cl2 (43 mg, 0.06 mmol). The mixture was heated to 70 °C and stirred for 2 h, then H2O was added and the mixture extracted with EtOAc (2 × 10 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give tert-butyl N-[(8S,14S)-21-[4-(methoxymethyl)pyridin-3-yl]-18,18-dimethyl-9,15-dioxo-4-[(triisopropylsilyl)oxy]-16-oxa-10,22,28-triazapentacyclo[18.5.2.1A[2,6].1A[10,14].0A[23,27]]nonacosa-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamate (370 mg, 74% yield) as a foam. LCMS (ESI): m/z [M+H] calculated for C48H67N5O7Si 853.6; found 854.6.

[00366] Step 3. A mixture of tert-butyl N-[(8S,14S)-21-[4-(methoxymethyl)pyridin-3-yl]-18,18-dimethyl-9,15-dioxo-4-[(tr-isopropylsilyl)oxy]-16-oxa- 10,22,28-triazapentacyclo[18.5.2.1A[2,6].1A[10,14].0A[23,27]]nonacosa- 1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamate (350 mg, 0.41 mmol), Cs2CO3 (267 mg, 0.82 mmol) and EtI (128 mg, 0.82 mmol) in DMF (4 mL) was stirred at 35°C overnight. H2O was added and the mixture was extracted with EtOAc (2 x 15 mL). The combined organic layers were washed with brine (15 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give tert-butyl N-[(8S,14S)-22-ethyl-21-[4-(methoxymethyl)pyridin-3-yl]-18,18-dimethyl-9,15-dioxo-4-[(triisopropylsilyl)oxy]-16-oxa-10,22,28-triazapentacyclo[18.5.2.1A[2,6].1A[10,14].0A[23,27]]nonacosa- 1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamate (350 mg, 97% yield) as an oil. LCMS (ESI): m/z [M+H] calculated for C50H71N5O7Si 881.5; found 882.6.

[00367] Step 4. A mixture of tert-butyl N-[(8S,14S)-22-ethyl-21-[4-(methoxymethyl)pyridin-3-yl]-18,18-dimethyl-9,15-dioxo-4-[(tr-isopropylsilyl)oxy]-16-oxa-10,22,28-triazapentacyclo[18.5.2.1A[2,6].1A[10,14].0A[23,27]]nonacosa-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamate (350 mg, 0.4 mmol) and 1M TBAF in THF (0.48 mL, 0.480 mmol) in THF (3 mL) at 0 °C under an Ar atmosphere was stirred for 1 h. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give tert-butyl N-[(8S,14S)-22-ethyl-4-hydroxy-21-[4-(methoxymethyl)pyridin-3-yl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.1A[2,6].1A[10,14].0A[23,27]]nonacosa-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamate (230 mg, 80% yield) as an oil. LCMS (ESI): m/z [M+H] calcd for C41H51N5O7 725.4; found 726.6.

[00368] Step 5. To a mixture of tert-butyl N-[(8S,14S)-22-ethyl-4-hydroxy-21-[4-(methoxymethyl)pyridin-3-yl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.1A[2,6].1A[10,14].0A[23,27]]nonacosa-1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]carbamate (200 mg, 0.28 mmol) in 1,4-dioxane (2 mL) at 0 °C under an Ar atmosphere was added 4 M HCl in 1,4-dioxane (2 mL, 8 mmol). The mixture was allowed to warm to rt and stirred overnight, then concentrated under reduced pressure to give (8S,14S)-8-amino-22-ethyl-4-hydroxy-21-[4-(methoxymethyl)pyridin-3-yl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.1A[2,6].1A[10,14].0A[23,27]]nonacosa- 1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione (200 mg). LCMS (ESI): m/z [M+H] calcd for C36H43N5O5 625.3; found 626.5.

[00369] Step 6. To a mixture of (2S)-3-methyl-2-[N-methyl-1-[(3S)-1-(prop-2-enoyl)pyrrolidin-3-yl]formamido]butanoic acid (108 mg, 0.38 mmol) and (8S,14S)-8-amino-22-ethyl-4-hydroxy-21-[4-(methoxymethyl)pyridin-3-yl]-18,18-dimethyl-16-oxa-10,22,28-triazapentacyclo[18.5.2.1A[2,6].1A[10,14].0A[23,27]]nonacosa- 1(26),2,4,6(29),20,23(27),24-heptaene-9,15-dione (200 mg, 0.32 mmol) in DCM (3 mL) at 0 °C was added DIPEA (165 mg, 1.3 mmol) and COMU (274 mg, 0.64 mmol) in portions. The mixture was stirred at 0 °C for 2 h, H2O added and extracted with EtOAc (2 × 10 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography then prep-HPLC to yield (2S)-N-[(8S,14S)-22-ethyl-4-hydroxy-21-[4-(methoxymethyl)pyridin-3-yl]-18,18-dimethyl-9,15-dioxo-16-oxa-10,22,28-triazapentacyclo[18.5.2.1A[2,6].1A[10,14].0A[23,27]]nonacosa- 1(26),2,4,6(29),20,23(27),24-heptaen-8-yl]-3-methyl-2-[N-methyl-1-[(3S)-1-(prop-2-enoyl)pyrrolidin-3-yl]formamido]butanamide (16 mg, 5.6% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C50H63N7O8 889.5; found 890.6; 1H NMR (400 MHz, DMSO-d6) δ 9.33 (dd, J = 9.1, 6.9 Hz, 1H), 8.79 - 8.46 (m, 2H), 7.93 (s, 1H), 7.68 - 7.58 (m, 2H), 7.53 (t, J = 8.5 Hz, 1H), 7.26 - 6.98 (m, 2H), 6.71 - 6.47 (m, 2H), 6.24 - 6.07 (m, 1H), 5.80 - 5.60 (m, 1H), 5.49 - 5.18 (m, 1H), 4.45 - 4.07 (m, 4H), 4.08 - 3.87 (m, 3H), 3.87 - 3.64 (m, 4H), 3.64 - 3.40 (m, 5H), 3.34 (s, 2H), 3.30 (s, 2H), 3.23 (d, J = 1.8 Hz, 1H), 2.94 - 2.74 (m, 6H), 2.16 - 2.01 (m, 3H), 1.82 - 1.47 (m, 3H), 1.08 (q, J = 8.9, 8.0 Hz, 1H), 1.00 - 0.88 (m, 6H), 0.82 (d, J = 10.8 Hz, 4H), 0.76 - 0.66 (m, 2H), 0.44 (d, J = 14.2 Hz, 3H). EXAMPLE A316. SYNTHESIS OF (2R)-2-(((1 -(4-(DIMETHYLAMINO)-4-METHYLPENT-2- INOYL)AZETIDIN-3-YL)OXY)METHYL)-N-((63S,4S)-11 -ETHYL-12-(2-((S)-1 - METHOXYETHYL)PYRIDIN-3-IL)-10,10-DIMETHYL-5,7-DIOXO-61,62,63,64,65,66-HEXA-HYDRO- 11H-8-OXA-1(5,3)-INDOLE-6(1,3)-PYRDAZINE-2(1,3)-BENZENOCYCLOUNDECAPHANE-4-YL)-3-METHYLBUTANAMIDE

[00370] Step 1. To a mixture of 2-(((1-((benzyloxy)carbonyl)azetidin-3-yl)oxy)methyl)-3-methylbutanoic acid (650 mg, 2 mmol) and tert-butyl dicarbonate (883 mg, 4 mmol) in tBuOH (10 mL) was added 4-dimethylaminopyridine (124 mg, 1 mmol). The mixture was heated to 30 °C and stirred for 1 h, then diluted with H2O (50 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to afford benzyl 3-(2-(tert-butoxycarbonyl)-3-methylbutoxy)azetidine-1-carboxylate (450 mg, 56% yield) as an oil. LCMS (ESI): m/z [M+Na] calculated for C21H31NO5Na 400.2; found 400.2.

[00371] Step 2. A mixture of benzyl 3-(2-(tert-butoxycarbonyl)-3-methylbutoxy)azetidine-1-carboxylate (450 mg, 1.19 mmol) and Pd/C (50 mg) in THF (30 mL) was stirred for 2 h under an atmosphere of H2 (15 psi). The mixture was filtered and the filtrate was concentrated under reduced pressure to give tert-butyl 2-((azetidin-3-yloxy)methyl)-3-methylbutanoate (300 mg, 100% yield) as an oil. LCMS (ESI): m/z [M+H] calcd for C13H26NO3 243.2; found 244.2; 1H NMR (400 MHz, CDCl3) δ 4.35 - 4.25 (m, 1H), 3.71 - 3.63 (m, 2H), 3.63 - 3.56 (m, 2H), 3.50 (t, J = 8.0 Hz, 1H), 3.43 (dd, J = 9.0, 4.0 Hz, 1H), 2.37 - 2.26 (m, 1H), 2.21 (br. s, 1H), 1.92 - 1.81 (m, 1H), 1.47 (s, 9H), 0.93 (d, J = 6.8 Hz, 6H).

[00372] Step 3. To a mixture of tert-butyl 2-((azetidin-3-yloxy)methyl)-3-methylbutanoate (270 mg, 1.11 mmol), 4-(dimethylamino)-4-methylpent-2-ynoic acid (860 mg, 5.55 mmol), and DIPEA (1.56 g, 11.1 mmol) in DMF (20 mL) at 0 °C was added T3P (2.12 g, 6.7 mmol). The mixture was stirred at 0 °C for 1 h, diluted with EtOAc (200 mL), then washed with H2O (30 mL × 5), brine (30 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give tert-butyl 2-(((1-(4-(dimethylamino)-4-methylpent-2-ynoyl)azetidin-3-yl)oxy)methyl)-3-methylbutanoate (200 mg, 47% yield) as an oil. LCMS (ESI): m/z [M+H] calcd for C21H36N2O4 380.3; found 381.3.

[00373] Step 4. To a mixture of tert-butyl 2-(((1-(4-(dimethylamino)-4-methylpent-2-ynoyl)azetidin-3-yl)oxy)methyl)-3-methylbutanoate (190 mg, 0.5 mmol) in DCM (4 mL) was added TFA (2 mL). The mixture was stirred for 1 h, then concentrated under reduced pressure to give 2-(((1-(4-(dimethylamino)-4-methylpent-2-ynoyl)azetidin-3-yl)oxy)methyl)-3-methylbutanoic acid (162 mg, 100% yield) as an oil, which was used directly in the next step without further purification. LCMS (ESI): m/z [M+H] calcd for C17H28N2O4 324.2; found 325.3.

[00374] Step 5. To a solution of (2S)-3-(3-bromophenyl)-2-[(tert-butoxycarbonyl)amino]propanoic acid (100 g, 290 mmol) in DMF (1 L) at room temperature was added NaHCO3 (48.8 g, 581.1 mmol) and MeI (61.9 g, 435.8 mmol). The reaction mixture was stirred for 16 h and was then quenched with H2O (1 L) and extracted with EtOAc (3 x 1 L). The combined organic layers were washed with brine (3 x 500 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (13% EtOAc/petroleum ether) to give methyl (S)-3-(3-bromophenyl)-2-((tert-butoxycarbonyl)amino)propanoate (109 g, crude). LCMS (ESI): m/z [M+Na] calculated for C15H20BrNO4 380.05; found 380.0.

[00375] Step 6. To a stirred solution of methyl (2S)-3-(3-bromophenyl)-2-[(tert-butoxycarbonyl)amino]propanoate (108 g, 301.5 mmol) and bis(pinacolato)diboron (99.53 g, 391.93 mmol) in 1,4-dioxane (3.2 L) was added KOAc (73.97 g, 753.70 mmol) and Pd(dppf)Cl2 (22.06 g, 30.15 mmol). The reaction mixture was heated at 90 °C for 3 h and was then cooled to room temperature and extracted with EtOAc (2 x 3 L). The combined organic layers were washed with brine (3 x 800 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (5% EtOAc/petroleum ether) to give methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanoate (96 g, 78.6% yield). LCMS (ESI): m/z [M+Na] calcd for C21H32BNO6 428.22; found 428.1.

[00376] Step 7. To a mixture of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]propanoate (94 g, 231.9 mmol) and 3-(5-bromo-1H-indol-3-yl)-2,2-dimethylpropyl acetate (75.19 g, 231.93 mmol) in 1,4-dioxane (1.5 L) and H2O (300 mL) was added K2CO3 (64.11 g, 463.85 mmol) and Pd(DtBPF)Cl2 (15.12 g, 23.19 mmol). The reaction mixture was heated to 70 °C and stirred for 4 h. The reaction mixture was extracted with EtOAc (2 × 2 L) and the combined organic layers were washed with brine (3 × 600 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (20% EtOAc/petroleum ether) to give (S)-3-(3-(3-(3-acetoxy-2,2-dimethylpropyl)-1H-indol-5-yl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoate (130 g, crude). LCMS (ESI): m/z [M+H] calcd for C30H38N2O6 523.28; found 523.1.

[00377] Step 8. To a solution of (2S)-3-(3-[3-[3-(acetyloxy)-2,2-dimethylpropyl]-1H-indol-5-yl]phenyl)-2-[(tert-butoxycarbonyl)amino]propanoate (95.0 g, 181.8 mmol) and iodine (36.91 g, 145.41 mmol) in THF (1 L) at -10 °C was added AgOTf (70.0 g, 272.7 mmol) and NaHCO3 (22.9 g, 272.65 mmol). The reaction mixture was stirred for 30 min and was then quenched by the addition of Na2S2O3 (100 mL) at 0 °C. The resulting mixture was extracted with EtOAc (3 × 1 L), and the combined organic layers were washed with brine (3 × 500 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (50% EtOAc/petroleum ether) to give methyl (S)-3-(3-(3-(3-acetoxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoate (49.3 g, 41.8% yield). LCMS (ESI) m/z: [M + H] calcd for C30H37IN2O6: 649.18; found 649.1.

[00378] Step 9: To a solution of (2S)-3-(3-[3-[3-(acetyloxy)-2,2-dimethylpropyl]-2-iodo-1H-indol-5-yl]phenyl)-2-[(tert-butoxycarbonyl)amino]propanoate (60 g, 92.5 mmol) in THF (600 mL) was added a solution of LiOH^O (19.41 g, 462.5 mmol) in H2O (460 mL). The resulting solution was stirred overnight and then the pH was adjusted to 6 with HCl (1 M). The resulting solution was extracted with EtOAc (2 x 500 mL) and the combined organic layers were washed with sat. brine. (2 x 500 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give (S)-2-((tert-butoxycarbonyl)amino)-3-(3-(3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl)phenyl)propanoic acid (45 g, 82.1% yield). LCMS (ESI): m/z [M+Na] calcd for C27H33IN2O6 615.13; found 615.1.

[00379] Step 10. To a solution of (2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl]phenyl]propanoic acid (30 g, 50.6 mmol) and methyl (3S)-1,2-diazinane-3-carboxylate (10.9 g, 75.9 mmol) in DCM (400 mL) was added NMM (40.97 g, 405.08 mmol), HOBT (2.05 g, 15.19 mmol), and EDCI (19.41 g, 101.27 mmol). The reaction mixture was stirred overnight and then the mixture was washed with NH4Cl (2 × 200 mL) and saturated brine (2 × 200 mL), and the mixture was dried over Na2SO4, filtered, and concentrated under reduced pressure to give methyl (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl)phenyl)propanoyl)hexahydropyridazine-3-carboxylate (14 g, 38.5% yield). LCMS (ESI): m/z [M+H] calcd for C33H43IN4O6 718.23; found 719.4.

[00380] Step 11. To a solution of methyl (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl)phenyl)propanoyl)hexahydropyridazine-3-carboxylate (92 g, 128.0 mmol) in THF (920 mL) at 0°C was added a solution of LiOH^O (26.86 g, 640.10 mmol) in H2O (640 mL). The reaction mixture was stirred for 2 h and was then concentrated under reduced pressure to give (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl)phenyl)propanoyl)hexahydropyridazine-3-carboxylic acid (90 g, crude). LCMS (ESI): m/z [M+H] calcd for C32H41IN4O6 705.22; found 705.1.

[00381] Step 12. To a solution of (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[3-[3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl]phenyl]propanoyl]-1,2-diazinane-3-carboxylic acid (90 g, 127.73 mmol) in DCM (10 L) at 0 °C was added HOBt (34.52 g, 255.46 mmol), DIPEA (330.17 g, 2554.62 mmol), and EDCI (367.29 g, 1915.96 mmol). The reaction mixture was stirred for 16 h and was then concentrated under reduced pressure. The mixture was extracted with DCM (2 × 2 L), and the combined organic layers were washed with brine (3 × 1 L), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (50% EtOAc/petroleum ether) to give tert-butyl ((63S,4S)-12-iodo-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (70 g, 79.8% yield). LCMS (ESI): m/z [M+H] calculated for C32H39IN4O5 687.21; found 687.1.

[00382] Step 13. A 1 L round bottom flask was charged with tert-butyl ((63S,4S)-12-iodo-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundecaphane-4-yl)carbamate (22.0 g, 32.042 mmol), toluene (300.0 mL), Pd2(dba)3 (3.52 g, 3.845 mmol), S-Phos (3.95 g, 9.613 mmol) and KOAc (9.43 g, 96.127 mmol) at room temperature. To the mixture was added 4,4,5,5-tetramethyl-1,3,2-dioxaborolane (26.66 g, 208.275 mmol) dropwise with stirring at room temperature. The resulting solution was stirred for 3 h at 60 °C. The resulting mixture was filtered and the filter cake was washed with EtOAc. The filtrate was concentrated under reduced pressure and the remaining residue was purified by silica gel column chromatography to give tert-butyl((63S,4S)-10,10-dimethyl-5,7-dioxo-12-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (22 g, 90% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C38H51BN4O7 687.3; found 687.4.

[00383] Step 14. A mixture of tert-butyl ((63S,4S)-10,10-dimethyl-5,7-dioxo-12-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-61,62,63,64,65,66- hexahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundephane-4-yl)carbamate(2.0 g, 2.8 mmol), 3-bromo-2-[(1S)-1-methoxyethyl]pyridine (0.60 g, 2.8 mmol), Pd(dppf)Cl2 (0.39 g, 0.5 mmol), and K3PO4 (1.2 g, 6.0 mmol) in English: 1,4-Dioxane (50 mL) and H2O (10 mL) under a N2 atmosphere was heated to 70 C and stirred for 2 h. The mixture was diluted with H2O (50 mL) and extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with brine (3 × 50 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give tert-butyl((63S,4S)-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (1.5 g, 74% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C40H49N5O6 695.4; found 696.5.

[00384] Step 15. To a solution of tert-butyl((63S,4S)-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (20 g, 28.7 mmol) and Cs2CO3 (18.7 g, 57.5 mmol) in DMF (150 mL) at 0 °C was added a solution of ethyl iodide (13.45 g, 86.22 mmol) in DMF (50 mL). The resulting mixture was stirred overnight at 35 °C and was then diluted with H2O (500 mL). The mixture was extracted with EtOAc (2 × 300 mL), and the combined organic layers were washed with brine (3 × 100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography to give tert-butyl((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (4.23 g, 18.8% yield) and the atropisomer (5.78 g, 25.7% yield) as solids. LCMS (ESI): m/z [M+H] calcd for C42H53N5O6 724.4; found 724.6.

[00385] Step 16. A mixture of tert-butyl ((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexa- hydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundephane-4-yl)carbamate (1.3 g, 1.7 mmol) in TFA (10 mL) and DCM (20 mL) was stirred at 0 C for 2 h. The mixture was concentrated under reduced pressure to afford (63S,4S)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-5,7-dione (1.30 g, crude) as a solid. LCMS (ESI): m/z [M+H] calcd for C37H45N5O4 623.3; found 624.4.

[00386] Step 17. To a mixture of (63S,4S)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H- 8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundephane-5,7-dione (258 mg, 0.41 mmol) and 2-(((1)-(4-(dimethylamino)-4-methylpent-2-inoyl)azetidin-3-yl)oxy)methyl)-3-methylbutanoic acid (162 mg, 0.5 mmol) in DMF (4 mL) at 0°C was added a mixture of HATU (188 mg, 0.5 mmol) and DIPEA (534 mg, 4.14 mmol) in DMF (2 mL). The mixture was stirred at 0 °C for 1 h, then diluted with H2O (30 mL) and extracted with EtOAc (30 mL × 3). The combined organic layers were concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 2-(((1-(4-(dimethylamino)-4-methylpent-2-ynoyl)azetidin-3-yl)oxy)methyl)-N-((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-1H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)-3-methylbutanamide (250 mg, 64% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C54H71N7O7 929.5; found 930.5; 1H NMR (400 MHz, CD3OD) δ 8.90 - 8.79 (m, 1H), 8.54 - 8.21 (m, 1H), 8.15 - 7.91 (m, 2H), 7.88 - 7.67 (m, 2H), 7.65 - 7.52 (m, 2H), 7.47 - 7.15 (m, 2H), 5.80 - 5.52 (m,1H), 4.53 - 4.23 (m, 5H), 4.23 - 3.93 (m, 3H), 3.90 - 3.76 (m, 2H), 3.75 - 3.58 (m,3H), 3.57 - 3.44 (m, 1H), 3.38 (s, 1H), 3.29 - 3.26 (m, 2H), 3.21 - 2.85 (m, 8H), 2.82 - 2.65 (m, 3H), 2.51 - 2.30 (m, 1H), 2.24 - 2.03 (m, 1H), 1.99 - 1.87 (m, 1H) 0.79 (m, 3H), 0.79 - 0.63 (m, 3H), 0.56 (s, 1H).

[00387] Step 18. 2-(((1-(4-(dimethylamino)-4-methylpent-2-inoyl)azetidin-3-yl)oxy)methyl)-N-((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)- 10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-1H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundephane-4-yl)-3-methylbutanamide (180mg, 0.194 mmol) was purified by prep-HPLC to generate (2R)-2-(((1-(4- (dimethylamino)-4-methylpent-2-inoyl)azetidin-3-yl)oxy)methyl)-N-((63S,4S)-11-ethyl-12- (2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexa- hydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundephane-4-yl)-3-methylbutanamide (41.8 mg, 23.2% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C54H71N7O7 930.5; found 930.5; 1H NMR (400 MHz, MeOD) δ 8.74 (d, J = 4.0 Hz, 1H), 8.53 - 8.30 (m, 1H), 8.10 - 7.95 (m, 1H), 7.94 - 7.80 (m, 2H), 7.68 (t, J = 8.0 Hz, 1H), 7.65-7.58 (m, 1H), 7.58-7.46(m, 2H), 7.38-7.17 (m, 2H), 5.73-5.60 (m, 1H), 4.52-4.40 (m, 1H), 4.35-4.15 (m, 4H), 4.14-3.95 (m, 2H), 3.90-3.72 (m, 3H), 3.71-3.45 (m, 4H), 3.30-3.20 (m, 3H), 3.06-2.72 (m, 5H), 2.49 - 2.28 (m, 4H), 2.28 - 2.20 (m, 3H), 2.18 - 2.06 (m, 1H),2.00 - 1.90 (m, 1H), 1.90 - 1.52 (m, 4H), 1.52 - 1.40 (m, 5H), 1.40 - 1.22 (m, 4H), 1.09 - 0.92 (m, 8H), 0.90 - 0.75 (m, 3H), 0.71 - 0.52 (m, 3H) and (2S)-2-(((1-(4-(dimethylamino)-4-methylpent-2-inoyl)azetidin-3-yl)oxy)methyl)-N-((63S,4S)-11-ethyl-12- (2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundecaphane- 4-yl)-3-methylbutanamide (51.2 mg, 28.4% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C54H71N7O7 930.5; found 930.3; 1H NMR (400 MHz, MeOD) δ 8.74 (d, J = 4.0 Hz, 1H), 8.28 - 8.20 (m, 0.6H), 8.11 - 7.98 (m, 1H), 7.97 - 7.80 (m, 2H), 7.73 - 7.48 (m, 4H), 7.46 - 7.36 (m, 0.4H), 7.33 - 7.26 (m, 1H), 7.25 - 7.13 (m, 1H), 5.79 - 5.66 (m, 1H), 4.54 - 4.43 (m, 1H), 4.42 - 4.01 (m, 7H), 3.90 - 3.75 (m, 2H), 3.73 - 3.48 (m, 4H), 3.27 - 3.12 (m, 3H), 3.08 - 2.99 (m, 1H), 2.96 - 2.85 (m, 2H), 2.84 - 2.69 (m, 2H), 2.69 - 2.49 (m, 6H), 2.41 - 2.29 (m, 1H), 2.15 - 2.05 (m, 1H), 1.95 - 1.85 (m, 1H), 1.84 - 1.71 (m, 1H), 1.71 - 1.38 (m, 11H), 1.14 - 1.00 (m, 3H), 1.00 - 0.71 (m, 9H), 0.70 - 0.56 (m, 3H).EXAMPLE A427. SYNTHESIS OF 3-((1-(4-(DIMETHYLAMINO)-4-METHYLPENT-2-INOYL)AZETIDIN-3-YL)OXI)- N -((63 S , 4 S, Z)-11-ETHYL-12-(2-(( S )-1-METHOXYETHYL)PYRIDIN- 3-IL)-10,10-DIMETHYL-5,7-DIOXO-61,62,63,64,65,66-HEXA-HYDRO-11 H-8-OXA-2(4,2)- TlAZOLA-1(5,3)-lNDOLA-6(1,3)-PlRIDAZINECYCLOUNDECAPHANE-4-lL)PROPANAMIDE

[00388] Step 1. To a mixture of tert-butyl prop-2-ynoate (5 g, 40 mmol) and [3-(3-hydroxyazetidin-1-yl)phenyl]methyl formate (4.1 g, 20 mmol) in DCM (150 mL) was added DMAP (9.8 g, 80 mmol). The mixture was stirred for 2 h, then diluted with H2O and washed with H2O (60 mL × 3). The organic layer was dried over Na2SO4, filtered, the filtrate was concentrated under reduced pressure, and the residue purified by silica gel column chromatography to give benzyl (E)-3-((3-(tert-butoxy)-3-oxoprop-1-en-1-yl)oxy)azetidine-1-carboxylate (6.6 g, 90% yield) as an oil. LCMS (ESI): m/z [M+Na] calcd for C18H23NO5Na 356.2; found 356.2.

[00389] Step 2. A mixture of benzyl (E)-3-((3-tert-butoxy)-3-oxoprop-1-en-1-yl)oxy)azetidine-1-carboxylate (1.4 g, 4 mmol) and Pd/C (200 mg) in THF (10 mL) was stirred under an H2 atmosphere (1 atmosphere) for 16 h. The mixture was filtered and the filtrate was concentrated under reduced pressure to give tert-butyl 3-(azetidin-3-yloxy)propanoate, which was used directly in the next step. LCMS (ESI): m/z [M+H] calculated for C10H19NO3 201.1; found 202.2.

[00390] Step 3. To a mixture of tert-butyl 3-(azetidin-3-yloxy)propanoate (300 mg, 1.5 mmol) and 4-(dimethylamino)-4-methylpent-2-ynoic acid (2.3 g, 15 mmol) in DMF (15 mL) at 5 °C was added DIPEA (1.9 g, 15 mmol) and T3P (4.77 g, 7.5 mmol) dropwise. The mixture was stirred at 5 °C for 2 h, then H2O and EtOAc (80 mL) were added. The organic and aqueous layers were separated, and the organic layer was washed with H2O (20 mL × 3), brine (30 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative HPLC to afford tert-butyl 3-((1-(4-(dimethylamino)-4-methylpent-2-ynoyl)azetidin-3-yl)oxy)propanoate (60 mg, 12% yield) as an oil. LCMS (ESI): m/z [M+H] calculated for C18H30N2O4 338.2; found 339.2.

[00391] Step 4. A mixture of tert-butyl 3-((1-(4-(dimethylamino)-4-methylpent-2-ynoyl)azetidin-3-yl)oxy)propanoate (70 mg, 0.21 mmol) in TFA/DCM (1:3, 2 mL) was stirred at 0-5 °C for 1 h, then concentrated under reduced pressure to give 3-({1-[4-(dimethylamino)-4-methylpent-2-ynoyl]azetidin-3-yl}oxy)propanoic acid (56 mg, 95% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C14H22N2O4 282.2; found 283.3.

[00392] Step 5. To a mixture of 3-((1-(4-(dimethylamino)-4-methylpent-2-inoyl)azetidin-3-yl)oxy)propanoic acid (56 mg, 0.19 mmol), (63S,4S,Z)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-thiazole-1(5,3)-indole- 6(1,3)-pyridazinecycloundefane-5,7-dione (90 mg, 0.14 mmol) and DIPEA (200 mg, 1.9 mmol) in DMF (1 mL) at 0°C was added HATU (110 mg, 0.38 mmol) was added portionwise. The mixture was stirred at 0 °C for 1 h, then H2O was added and the mixture was extracted with EtOAx (150 mL × 2). The combined organic layers were washed with H2O (150 mL) and brine (150 mL), then dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by preparative HPLC to obtain 3-((1-(4-(dimethylamino)-4-methylpent-2-ynoyl)azetidin-3-yl)oxy)-N-((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-thiazole-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)propanamide (12.6 mg, 7.5% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C48H62N8O7S 894.5; found 895.3; 1H NMR (400 MHz, CD3OD) δ 8.73 (dd, J = 4.8, 1.6 Hz, 1H), 8.57 (s, 1H), 8.28 (s, 0.3H), 7.84 (m, 1H), 7.71 (m, 1H), 7.52 (m, 3H), 5.76 (dd, J = 30.2, 7.8 Hz, 1H), 4.40 (m, 4H), 4.32 - 4.12 (m, 4H), 4.06 (dd, J = 12.4, 6.0 Hz, 1H), 3.97 - 3.86 (m, 1H), 3.79 - 3.66 (m, 4H), 3.46 (dd, J = 14.8, 4.8 Hz, 1H), 3.41 - 3.33 (m, 3H), 3.29 - 3.19 (m, 1H), 3.17 - 3.05 (m, 1H), 2.79 (m, 1H), 2.73 - 2.50 (m, 3H), 2.49 - 2.43 (m, 3H), 2.38 (s, 3H), 2.21 (dd, J = 12.6, 9.6 Hz, 1H), 1.95 (d, J = 12.8 Hz, 1H), 1.86 - 1.73 (m, 1H), 1.61 (dd, J = 12.6, 3.6 Hz, 1H), 1.51 (s, 2H), 1.46 - 1.43 (m, 4H), 1.38 - 1.27 (m, 3H), 1.01 - 0.86 (m, 6H), 0.44 (d, J = 11.6 Hz, 3H). EXAMPLE A716. SYNTHESIS OF (3S)-1 -ACRYLOYL-N-((2S)-1 -(((63S,4S)-11 -ETHYL- 12-(2-((S)-1 -METHOXYETHYL)PYRIDIN-3-IL)-1 0,10-DIMETHYL-5,7-DIOXO-21,22,23,26,61 ,62,63,64,65,66-DECA-HYDRO-11H-8-OXA-1 (5,3)-INDOLE-6(1,3)- PYRDAZINE-2(5,1)-PYRIDINECYCLOUNDECAPHANE-4-YL)AMINO)-3-METHYL-1-OXOBUTAN-2-YL)-N-METHYLPYRROLIDINE-3-CARBOXAMIDE

[00393] Step 1. To a solution of methyl (tert-butoxycarbonyl)-L-serinate (10 g, 45 mmol) in anhydrous MeCN (150 mL) was added DIPEA (17 g, 137 mmol). The reaction mixture was stirred at 45 °C for 2 h to give methyl 2-((tert-butoxycarbonyl)amino)acrylate in solution. LCMS (ESI): m/z [M+Na] calculated for C9H15NO4 201.1; found 224.1.

[00394] Step 2. To a solution of methyl 2-((tert-butoxycarbonyl)amino)acrylate (12 g, 60 mmol) in anhydrous MeCN (150 mL) at 0 °C was added 4-DMAP (13 g, 90 mmol) and (Boc)2O (26 g, 120 mmol). The reaction was stirred for 6 h, then quenched with H2O (100 mL) and extracted with DCM (200 mL x 3). The combined organic layers were washed with brine (150 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give methyl 2-(bis(tert-butoxycarbonyl)amino)acrylate (12.5 g, 65% yield) as a solid. LCMS (ESI): m/z [M+Na] calculated for C14H23NO6 301.2; found 324.1.

[00395] Step 3. To a mixture of 5-bromo-1,2,3,6-tetrahydropyridine (8.0 g, 49 mmol) in MeOH (120 mL) under an Ar atmosphere was added methyl 2-{bis[(tert-butoxy)carbonyl]amino}prop-2-enoate (22 g, 74 mmol). The mixture was stirred for 16 h, then concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give methyl 2-(bis(tert-butoxycarbonyl)amino)-3-(5-bromo-3,6-dihydropyridin-1(2H)-yl)propanoate (12 g, 47% yield) as an oil. LCMS (ESI): m/z [M+H] calculated for C19H31BrN2O6 462.1; found 463.1.

[00396] Step 4. To a mixture of methyl 2-(bis(tert-butoxycarbonyl)amino)-3-(5-bromo-3,6-dihydropyridin-1(2H)-yl)propanoate (14 g, 30 mmol) in 1,4-dioxane (30 mL) and H2O (12 mL) was added LiOH (3.6 g, 151 mmol). The mixture was heated to 35 °C and stirred for 12 h, then 1 M HCl was added and the pH was adjusted to ~3-4. The mixture was extracted with DCM (300 mL x 2), and the combined organic layers were dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to obtain 3-(5-bromo-3,6-dihydropyridin-1(2H)-yl)-2-((tert-butoxycarbonyl)amino)propanoic acid (10 g, 85% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C13H21BrN2O4 348.1; found 349.0.

[00397] Step 5. To a mixture of 3-(5-bromo-3,6-dihydropyridin-1(2H)-yl)-2-((tert-butoxycarbonyl)amino)propanoic acid (10 g, 30 mmol), DIPEA (12 g, 93 mmol), and methyl (3S)-1,2-diazinane-3-carboxylate (5.4 g, 37 mmol) in DMF (100 mL) at 0 °C under an Ar atmosphere was added HATU (13 g, 34 mmol). The mixture was stirred at 0 °C for 2 h, then H2O was added and the mixture was extracted with EtOAc (300 mL × 2). The combined organic layers were dried over anhydrous Na2SO4, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by preparative HPLC to give methyl (3S)-1-(3-(5-bromo-3,6-dihydropyridin-1(2H)-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate (9.0 g, 55% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C19H31BrN4O5 474.1; found 475.1.

[00398] Step 6. A mixture of methyl (3S)-1-(3-(5-bromo-3,6-dihydropyridin-1(2H)-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate (9.0 g, 18 mmol), K2CO3 (4.5 g, 32 mmol), Pd(dppf)Cl2.DCM (1.4 g, 2 mmol), 3-(1-ethyl-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indol-3-yl)-2,2-dimethylpropan-1-ol (9.8g, 20 mmol) in 1,4-dioxane (90 mL) and H2O (10 mL) under an Ar atmosphere was heated to 75 °C and stirred for 2 h. H2O was added and the mixture was extracted with EtOAc (200 mL x 3). The combined organic layers were dried over Na2SO4, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain methyl (3S)-1-(2-((tert-butoxycarbonyl)amino)-3-(5-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)-3,6-dihydropyridin-1(2H)-yl)propanoyl)hexahydropyridazine-3-carboxylate (4.0 g, 25% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C42H60N6O7 760.5; found 761.4.

[00399] Step 7. To a mixture of methyl (3S)-1-(2-((tert-butoxycarbonyl)amino)-3-(5-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)-3,6-dihydropyridin-1(2H)-yl)propanoyl)hexahydropyridazine-3-carboxylate (4.1 g, 5.0 mmol) in THF (35 mL) at 0 °C was added LiOH (0.60 g, 27 mmol). The mixture was stirred at 0 °C for 1.5 h, then 1 M HCl was added to adjust the pH to ~6-7, and the mixture was extracted with EtOAc (200 mL × 3). The combined organic layers were dried over Na2SO4, filtered, and filtrate was concentrated under reduced pressure to give (3S)-1-(2-((tert-butoxycarbonyl)amino)-3-(5-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)-3,6-dihydropyridin-1(2H)-yl)propanoyl)hexahydropyridazine-3-carboxylic acid (3.6 g, 80% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C41H58N6O7 746.4; found 747.4.

[00400] Step 8. To a mixture of acid (3S)-1-(2-((tert-butoxycarbonyl)amino)-3-(5-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1- methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)-3,6-dihydropyridin-1(2H)-yl)propanoyl)hexahydropyridazine-3-carboxylic acid (3.6 g, 5.0 mmol) and DIPEA (24 g, 190 mmol) in DCM (700 mL) under an Ar atmosphere was added EDCI.HCl (28 g, 140 mmol) and HOBT (6.5 g, 50 mmol). The mixture was heated to 30 °C and stirred for 16 h at 30 °C, then concentrated under reduced pressure. The residue was diluted with EtOAc (200 mL) and washed with H2O (200 mL × 2), brine (200 mL), dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to obtain tert-butyl((63S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-21,22,23,26,61,62,63,64,65,66-decahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(5,1)-pyridinecycloundecaphane-4-yl)carbamate (1.45 g, 40% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C41H56N6O6 728.4; found 729.4.

[00401] Step 9. To a mixture of tert-butyl ((63S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo- 21,22,23,26,61,62,63,64,65,66-decahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(5,1)-pyridinecycloundecaphane-4-yl)carbamate (130 mg, 0.20 mmol) in DCM (1.0 mL) at 0 C TFA (0.3 mL) was added. The mixture was warmed to room temperature and stirred for 2 h, then concentrated under reduced pressure to give (63S)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-21,22,23,26,61,62,63,64,65,66-decahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(5,1)-pyridinecycloundecaphane-5,7-dione, which was used directly in the next step without further purification. LCMS (ESI): m/z [M+H] calcd for C36H48N6O4 628.4; found 629.4.

[00402] Step 10. To a mixture of ((63S)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-21,22,23,26,61,62,63,64,65,66-deca- hydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(5,1)-pyridinecycloundephane-5,7-dione (130 mg, 0.2 mmol), DIPEA (270 mg, 2.0 mmol) and (2S)-3-methyl-acid 2-{N-methyl-1-[(3S)-1-(prop-2-enoyl)pyrrolidin-3-yl]formamido}butanoic acid (118 mg, 0.40 mmol) in DMF (3.0 mL) at 0 °C under an Ar atmosphere was added HATU (87 mg, 0.30 mmol) in portions. The mixture was stirred at 0 °C for 1 h, then diluted with H 2 O extracted with EtOAc (30 mL × 2). The combined organic layers were dried over anhydrous Na 2 SO 4 , filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by preparative HPLC to give (3S)-1-acryloyl-N-((2S)-1-(((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)- 10,10-Dimethyl-5,7-dioxo-21,22,23,26,61,62,63,64,65,66-decahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(5,1)-pyridine(cycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)-N-methylpyrrolidine-3-carboxamide (17.2 mg, 10% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C50H68N6O4 892.5; found 893.5;1H NMR (400 MHz, CD3OD) δ 8.74 (d, J = 4.4 Hz, 1H), 7.93 - 7.90 (m, 1H), 7.56 - 7.51 (m, 3H), 7.43 (d, J = 4.4 Hz, 1H), 6.63 - 6.53 (m, 2H), 6.33 - 6.23 (m, 2H), 5.83 - 5.70 (m, 1H), 4.73 - 4.70 (d, J = 11.0 Hz, 1H), 4.48 - 4.45 (d, J = 13.0 Hz, 1H), 4.12 - 4.10 (m, 3H), 3.86 - 3.81 (m, 4H), 3.79 - 3.75 (m,1H), 3.72 - 3.69 (m, 3H), 3.57 - 3.47 (m, 2H), 3.21 - 3.09 (m, 1H), 3.07 - 3.04 (q,4H), 3.02 - 2.95 (m, 3H), 2.86 - 2.82 (m, 3H), 2.66 - 2.48 (m, 2H), 2.29 - 2.17 (m,4H), 2.11 - 1.98 (m, 2H), 1.95 - 1.91 (m,1H), 1.45 (d, J = 6.2 Hz, 3H), 1.23 - 1.16 (m, 2H), 1.09 - 1.04 (m, 1H), 0.97 - 0.93 (m, 3H), 0.92 - 0.81 (m, 5H), 0.67 - 0.63 (m, 3H).EXAMPLE A663. SYNTHESIS OF (2R)-2-(((1-(4-(DIMETHYLAMINO)-4-METHYLPENT-2- INOYL)386IRIDINE3863-IL)OXY)METHYL)-N-((63S,4S)-11 -ETHYL-12-(2-((S)-1 - METHOXYETHYL)386IRIDINE-3-IL)-10,10-DIMETHYL-5,7-DIOXO-21,22,23,26,61 ,62,63,64,65,66-DECA-HYDRO-11H-8-OXA-1 (5,3)-INDOLE-6(1,3)- PYRDAZINE-2(5,1)-PYRIDINECYCLOUNDECAPHANE-4-YL)-3-METHYLBUTANAMIDE

[00403] To a mixture of acid (63S,4S)-4-amino-11-ethyl-12- (2-((S)-1-methoxyethyl)386pyridin-3-yl)-10,10-dimethyl-21,22,23,26,61,62,63,64,65,66-deca- hydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(5,1)-pyridinecycloundephane-5,7-dione (100 mg, 0.16 mmol), ®-2-(((1-(4-(dimethylamino)-4-methylpent-2-inoyl)386pyridine386-3-yl)oxy)methyl)-3-methylbutanoic acid (80 mg, 0.24 mmol) and DIPEA (825 mg, 6.4 mmol) in DMF (2 mL) at 0 C, HATU (95 mg, 0.24 mmol) was added. The reaction mixture was stirred at 0 °C for 1 h, then poured into H2O (60 mL), extracted with EtOAc (80 mL × 2). The combined organic layers were washed with H2O (80 mL) and brine (80 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography to give (2R)-2-(((1-(4-(dimethylamino)-4-methylpent-2-ynoyl)386pyridine386-3-yl)oxy)methyl)-N-((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)386pyridine-3-yl)-10,10-dimethyl-5,7-dioxo-21,22,23,26,61,62,63,64,65,66-decahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(5,1)-pyridinecycloundecaphane-4-yl)-3-methylbutanamide (55 mg, 36% yield) as solid. 1H NMR (400 MHz, CD3OD) δ 8.76 - 8.70 (m, 1H), 8.49 (dd, J = 4.3, 1.4 Hz, 0.1H), 7.93 - 7.87 (m, 1H), 7.58 - 7.50 (m, 3H), 7.41 (dd, J = 8.8, 3.2Hz, 1H), 6.26 (d, J = 16.8Hz, 1H), 5.96 (t, J = 9.6Hz, 1H), 4.47 (d, J = 12.8Hz, 1H), 4.39 - 4.28 (m, 2H), 4.21 - 3.97 (m, 5H), 3.96 - 3.70 (m, 5H), 3.68 - 3.54 (m, 3H), 3.51 - 3.35 (m, 1H), 3.11 (d, J = 22.7 Hz, 3H), 3.00 - 2.67 (m, 5H), 2.46 - 2.30 (m, 7H), 2.24 (s, 3H), 2.11 (d, J = 12.4 Hz, 1H), 1.92 (d, J = 13.2 Hz, 1H), 1.85 - 1.60 (m, 3H), 1.45 (d, J = 7.8 Hz, 6H), 1.32 (d, J = 16.0 Hz, 3H), 1.12 (dt, J = 24.5, 6.8Hz, 3H), 0.95 (m, 6H), 0.76 (m, 6H). LCMS (ESI): m/z [M+H] calcd for C53H74N8O7 934.6; found 935.5.EXAMPLEA646. SYNTHESIS OF (2R)-2-(((1-(4-(DIMETHYLAMINO)-4-METHYLPENT-2- INOYL)AZETIDIN-3-YL)OXY)METHYL)-N-((63S,4S)-11 -ETHYL-12-(2-((S)-1 - METHOXYETHYL)PYRIDIN-3-IL)-10,10-DIMETHYL-5,7-DIOXO-61,62,63,64,65,66-HEXA-HYDRO- 11H-8-OXA-1(5,3)-INDOLE-6(1,3)-PYRDAZINE-2(3,1)-PIPERIDINECYCLOUNDECAPHANE-4-YL)-3-METHYLBUTANAMIDE

[00404] Step 1. A mixture of tert-butyl ((63S)-11-ethyl-12-(2- ((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-21,22,23,26,61,62,63,64,65,66- decahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(5,1)-pyridinecycloundephane-4-yl)carbamate (0.2 g, 0.28 mmol) and Pd/C (0.2 g, 2 mmol) in MeOH (10 mL) was stirred at 25 C for 16 h under an H2 atmosphere. The reaction mixture was filtered through Celite, concentrated under reduced pressure to afford tert-butyl((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(3,1)-piperidinecycloundecaphane-4-yl)carbamate as solid. LCMS (ESI): m/z [M+H] calcd for C41H58N6O6 730.4; found 731.4.

[00405] Step 2. To a solution of tert-butyl((63S,4S)-11-ethyl- 12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(3,1)-piperidinecycloundecaphane-4-yl)carbamate (150 mg, 0.2 mmol) in DCM (1.5 mL) at 0 °C was added TFA (0.5 mL). The reaction mixture was stirred at 20 °C for 1 h, then concentrated under reduced pressure to give (63S,4S)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(3,1)-piperidinecycloundecaphane-5,7-dione as a solid. LCMS (ESI): m/z [M+H] calcd for C36H50N6O4 630.4; found 631.4.

[00406] Step 3. A mixture of (63S,4S)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8- oxa-1(5,3)-indole-6(1,3)-pyridazine-2(3,1)-piperidinecycloundecaphane-5,7-dione (240 mg, 0.4 mmol), DIPEA (982 mg, 2 mmol) and acid (R)-2-(((1-(4-(dimethylamino)-4-methylpent-2-inoyl)azetidin-3-yl)oxy)methyl)-3-methylbutanoic acid (148 mg, 0.45 mmol) in DMF (4 mL) at At 0 °C under an argon atmosphere, HATU (173 mg, 0.46 mmol) was added portionwise. The reaction mixture was stirred at 0 °C under an argon atmosphere for 1 h, then quenched with H2O at 0 °C. The resulting mixture was extracted with EtOAc (30 mL × 2). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by reversed-phase chromatography to give (2R)-2-(((1-(4-(dimethylamino)-4-methylpent-2-inoyl)azetidin-3-yl)oxy)methyl)-N-((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(3,1)-piperidinecycloundecaphane-4-yl)-3-methylbutanamide (150 mg, 38% yield) as solid. 1H NMR (400 MHz, CD3OD) δ 8.72 (d, J = 4.8 Hz, 1H), 7.88 (d, J = 8.0 Hz, 1H), 7.53-7.49 (m, 2H), 7.42 (d, J = 8.4 Hz, 1H), 7.18 (d, J = 8.4 Hz, 1H) (m, 2H), 3.55-3.50(m, 2H), 3.39-3.36 (m, 2H), 3.20 (s, 3H), 3.02 (s, 3H), 2.89-2.79 (m, 3H), 2.62-2.50 (m, 2H), 2.36 (s, 3H), 2.35-2.30 (m, 1H), 2.26(s, 3H), 2.20-1.15 (m, 1H), 1.971.93 (m, 3H), 1.81-1.76 (m, 4H), 1.64-1.61 (m, 2H), 1.46-1.43 (m, 6H), 1.36 (d, J = 14.8 Hz, 3H), 1.02 (s, 3H), 0.94 (m, 6H), 0.81 (s, 3H), 0.65 (s, 3H). LCMS (ESI): m/z [M+H] calculated for C53H76N8O7 936.6; found 937.5. Example A740. Synthesis of (3S)-1-acryloyl-N-((2S)-1-(((23S,63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo- 61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(3,1)-piperidinecycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)-N-methylpyrrolidine-3-carboxamide

[00407] A mixture of (63S,4S)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa- 1(5,3)-indole-6(1,3)-pyridazine-2(3,1)-piperidinecycloundecaphane-5,7-dione (140 mg, 0.20 mmol), DIPEA (570 mg, 4.4 mmol) and (2S)-3-methyl-2-{N-methyl-1- acid [(3S)-1-(prop-2-enoyl)pyrrolidin-3-yl]formamido}butanoic acid (124 mg, 0.40 mmol) in DMF (3.0 mL) at 0 C under an Ar atmosphere was added HATU (100 mg, 0.30 mmol) in portions. The mixture was stirred at 0 °C for 1 h, then H2O was added and the mixture was extracted with EtOAc (2 × 30 mL). The combined organic layers were dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by preparative HPLC to give (3S)-1-acryloyl-N-((2S)-1-(((23S,63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(3,1)-piperidinecycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)-N-methylpyrrolidine-3-carboxamide (41 mg, 20% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C50H70N8O7 894.5; found 895.5; 1H NMR (400 MHz, CD3OD) δ 8.72 (d, J = 4.8 Hz, 1H), 7.87 (d, J = 7.6 Hz, 1H), 7.51 - 7.49 (m, 2H), 7.42 - 7.37 (m, 1H), 7.18 - 7.14 (m, 1H), 6.64 - 6.54 (m, 1H), 6.30 - 6.23 (m, 1H), 5.77 - 5.70 (m, 2H), 4.65 - 4.60 (m, 1H), 4.50 - 4.40 (m, 1H), 4.27 - 4.16 (m, 2H), 4.00 - 3.95 (m, 2H), 3.83 - 3.78(m,2H), 3.73 - 3.60 (m, 4H), 3.51 - 3.36 (m, 3H), 3.22 - 3.19 (m, 4H), 3.07 (d, J = 6.8Hz, 2H), 2.99 (d, J = 12.0 Hz, 3H), 2.90 - 2.78 (m, 2H), 2.75 - 2.64 (m, 3H), 2.20 - 2.10 (m, 4H), 2.02 - 1.93 (m, 3H), 1.87 - 1.64 (m, 4H), 1.45 (d, J = 4.8 Hz, 3H), 1.06 - 1.00 (m, 4H), 0.97 - 0.89 (m, 3H), 0.83 - 0.79 (m, 3H), 0.66 (s, 3H).EXAMPLE A534. (2S)-2-((S)-7-(4-(DIMETHYLAMINO)-4-METHYLPENT-2-INOYL)-1 - oxo-2,7-DiAZAESPiRor4.4]NONAN-2-iL)-N-((63S,4S)-11 -ETHYL-12-(2-((S)-1 -METHOXYETHYL)PYRIDIN-3-IL)-10,10-DIMETHYL-5,7-DIOXO-61,62,63,64,65,66-HEXA-HYDRO- 11H-8-OXA-1(5,3)-INDOLE-6(1,3)-PYRDAZINE-2(1,3)-BENZENACYCLOUNDECAPHANE-4- IL)-3-METHYLBUTANAMIDE

[00408] Step 1. To a mixture of 1-tert-butyl 3-methyl pyrrolidine-1,3-dicarboxylate (20.0 g, 87.2 mmol) in THF (150 mL) at -78 °C under a nitrogen atmosphere was added 1 M LiHMDS in THF (113.4 mL, 113.4 mmol). After stirring at -78 °C for 40 min, allyl bromide (13.72 g, 113.4 mmol) was added and the mixture was allowed to warm to room temperature and stirred for 4 h. The mixture was cooled to 0 °C, saturated NaCl (30 mL) was added, and the mixture was extracted with EtOAc. The combined organic layers were dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give 1-(tert-butyl)3-methyl 3-allylpyrrolidine-1,3-dicarboxylate (17 g, 72% yield) as an oil. 1H NMR (300 MHz, CDCl3) δ 5.80 - 5.60 (m, 1H), 5.16 - 5.02 (m, 2H), 3.71 (s, 4H), 3.42 (d, J = 9.3 Hz, 2H), 3.27 (t, J = 11.2 Hz, 1H), 2.42 (d, J = 7.6 Hz, 2H), 2.38 - 2.24 (m, 1H), 2.05 (s, 1H), 1.85 (dt, J = 14.3, 7.5 Hz, 1H), 1.46 (s, 10H), 1.27 (t, J = 7.1 Hz, 1H).

[00409] Step 2. To a mixture of 1-(tert-butyl)3-methyl 3-allylpyrrolidine-1,3-dicarboxylate (4.0 g, 14.9 mmol) and 2,6-dimethylpyridine (3.18 g, 29.7 mmol) in 1,4-dioxane (200 mL) and H2O (100 mL) at 0 °C was added K2OsO4 2H2O (0.11 g, 0.3 mmol) in portions. The mixture was stirred for 15 min at 0 °C, then NaIO4 (6.35 g, 29.7 mmol) was added in portions. The mixture was stirred at room temperature for 3 h at room temperature, then cooled to 0 °C and saturated aqueous Na2S2O3 (50 mL) added. The mixture was extracted with EtOAc (3 × 100 mL), and the combined organic layers were washed with 2 M HCl, then dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain 1-(tert-butyl)3-methyl 3-(2-oxoethyl)pyrrolidine-1,3-dicarboxylate (4 g, 52% yield) as an oil. 1H NMR (300 MHz, CDCl3) δ 5.80 - 5.60 (m, 1H), 5.16 - 5.04 (m, 2H), 3.72 (s, 3H), 3.41 (s, 3H), 3.28 (d, J = 11.0 Hz, 1H), 2.44 (s, 2H), 2.31 (d, J = 9.1 Hz, 1H), 1.85 (dt, J = 12.7, 7.5 Hz, 1H), 1.69 (s, 1H), 1.47 (s, 10H).

[00410] Step 3. To a mixture of 1-(tert-butyl)3-methyl 3-(2-oxoethyl)pyrrolidine-1,3-dicarboxylate (6.30 g, 23.2 mmol) in MeOH (70 mL) at 0 °C was added benzyl (2S)-2-amino-3-methylbutanoate (7.22 g, 34.8 mmol) and ZnCl2 (4.75 g, 34.8 mmol). The mixture was warmed to room temperature and stirred for 30 min, then cooled to 0 °C and NaCNBH3 (2.92 g, 46.4 mmol) was added portionwise. The mixture was warmed to room temperature and stirred for 2 h, then cooled to 0 °C and saturated aqueous NH4Cl added. The mixture was extracted with EtOAc (3 × 200 mL), and the combined organic layers were washed with brine (150 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 1-(tert-butyl)3-methyl 3-(2-(((S)-1-(benzyloxy)-3-methyl-1-oxobutan-2-yl)amino)ethyl)pyrrolidine-1,3-dicarboxylate (6.4 g, 54% yield) as an oil. LCMS (ESI): m/z [M+H] calcd for C25H38N2O6 462.3; found 463.4.

[00411] Step 4. To a mixture of 1-(tert-butyl) 3-methyl 3-(2-(((S)-1-(benzyloxy)-3-methyl-1-oxobutan-2-yl)amino)ethyl)pyrrolidine-1,3-dicarboxylate (4.50 g, 9.7 mmol) in toluene (50 mL) was added DIPEA (12.57 g, 97.3 mmol) and DMAP (1.19 g, 9.7 mmol). The resulting mixture was heated to 80 °C and stirred for 24 h, then concentrated under reduced pressure and the residue was purified by preparative HPLC, then by chiral HPLC to give tert-butyl (R)-7-((S)-1-(benzyloxy)-3-methyl-1-oxobutan-2-yl)-6-oxo-2,7-diazaspiro[4.4]nonane-2-carboxylate (1.0 g, 32% yield) and tert-butyl (S)-7-((S)-1-(benzyloxy)-3-methyl-1-oxobutan-2-yl)-6-oxo-2,7-diazaspiro[4.4]nonane-2-carboxylate (1.0 g, 32% yield) and as an oil. LCMS (ESI): m/z [M+H] calculated for C24H34N2O5 430.5; found 431.2 and LCMS (ESI): m/z [M+H] calculated C24H34N2O5 430.3; found 431.2.

[00412] Step 5. A mixture of tert-butyl (R)-7-((S)-1-(benzyloxy)-3-methyl-1-oxobutan-2-yl)-6-oxo-2,7-diazaspiro[4.4]nonan-2-carboxylate (4.0 g) and 10% Pd/C (1 g) in MeOH (40 mL) was stirred at room temperature under an atmosphere of H2. The mixture was filtered through a pad of Celite and the filtrate was concentrated under reduced pressure to give (S)-2-((R)-7-(tert-butoxycarbonyl)-1-oxo-2,7-diazaspiro[4.4]nonan-2-yl)-3-methylbutanoic acid (4.9 g) as a solid. LCMS (ESI): m/z [M-H] calculated for C17H28N2O5 340.2; found 339.3.

[00413] Step 6. To a mixture of (63S,4S)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-5,7-dione (500 mg, 0.8 mmol) in DCM at 0 °C was added DIPEA (829 mg, 6.4 mmol), ((S)-2-((R)-7-(tert-butoxycarbonyl)-1-oxo-2,7-diazaspiro[4.4]nonan-2-yl)-3-methylbutanoic acid (273 mg, 0.8 mmol) and HATU (396 mg, 1.0 mmol) portionwise over 1 min. The mixture was allowed to warm to room temperature and stirred 2 h, then concentrated under reduced pressure and the residue was purified by preparative TLC to give tert-butyl (5R)-7-((2S)-1-(((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro- 11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl) -6-oxo-2,7-diazaspiro[4.4]nonane-2-carboxylate (500 mg, 64% yield) as an oil. LCMS (ESI): m/z [M+H] calculated for C54H71N7O8 945.5; found 946.5.

[00414] Step 7. To a mixture of tert-butyl (5R)-7-((2S)-1-(((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo- 61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)-6-oxo-2,7- diazaspiro[4.4]nonane-2-carboxylate (1.0 g, 1.06 mmol) in DCM (10 mL) at 0°C was added TFA (3 mL) dropwise. The mixture was warmed to room temperature and stirred for 1 h, then concentrated under reduced pressure to give (2S)-N-((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)-3-methyl-2-((S)-1-oxo-2,7-diazaspiro[4.4]nonan-2-yl)butanamide (1.3 g). LCMS (ESI): m/z [M-H] calcd for C49H63N7O6 846.1; found 845.5.

[00415] Step 8. To a mixture of (2S)-N-((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexa- hydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundephane-4-yl)-3-methyl-2-((S)-1-oxo-2,7-diazaspiro[4.4]nonan-2-yl)butanamide (500 mg, 0.59 mmol) and DIPEA (764 mg, 5.9 mmol) in DMF (5 mL) at 0°C acid was added English: 4-(dimethylamino)-4-methylpent-2-ynoic acid (110 mg, 0.71 mmol) and HATU (292 mg, 0.77 mmol) were added portionwise. The mixture was warmed to room temperature and stirred for 1 h, then H2O (10 mL) was added and the mixture was extracted with EtOAc (10 mL × 3). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by preparative HPLC to obtain (2S)-2-((S)-7-(4-(dimethylamino)-4-methylpent-2-ynoyl)-1-oxo-2,7-diazaspiro[4.4]nonan-2-yl)-N-((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)-3-methylbutanamide (177 mg, 28.94% yield). as a white solid. LCMS (ESI): m/z [M+H] calcd for C57H74N8O7 982.6; found 983.8; - 7.58 (m, 3H), 7.53 (dd, J = 7.7, 4.8 Hz, 1H), 7.24 (t, J = 7.6 Hz, 1H), 7.12 (d, J = 7.6 Hz, 1H), 5.32 (d, J = 9.7 Hz, 2H), 4.33 - 4.20 (m, 4H), 4.03 (dd, J = 15.0, 8.6 Hz, 2H), 3.88 - 3.82 (m, 1H), 3.63 (dq, J = 20.5, 10.3 Hz, 4H), 3.42 - 3.34 (m, 2H), 3.21 (s, 1H), 3.13 (d, J = 2.8 Hz, 3H), 2.87 (s, 2H), 2.83 - 2.72 (m, 2H), 2.69 - 2.62 (m, 1H), 2.21 (d, J = 22.6 Hz, 6H), 2.12 - 1.76 (m, 7H), 1.75 - 1.47 (m, 2H), 1.46 - 1.28 (m, 9H), 0.99 - 0.89 (m, 6H), 0.79 - 0.71 (m, 6H), 0.52 (s, 3H).EXAMPLE A341. SYNTHESIS OF (3S)-1 -ACRYLOYL-N-((2S)-1 -(((63S,4S)-25- (DIFLUOROMETHYL)-11-ETHYL-12-(2-((S)-1-METHOXYETHYL)PYRIDIN-3-IL)-10,10-DIMETHYL-5,7-DIOXO-61 ,62,63,64,65,66-HEXA-HYDRO-11H-8-OXA-1 (5,3)-INDOLE-6(1,3)-PYRIDAZINE-2(1,3)-BENZENACYCLOUNDECAPHANE-4-YL)AMINO)-3-METHYL-1-OXOBUTAN-2-YL) -N-METHYLPYRROLIDINE-3-CARBOXAMIDE

[00416] Step 1. To a mixture of 3-bromo-5-iodobenzaldehyde (4.34 g, 14.0 mmol) in DCM at 0 °C under a N 2 atmosphere was added BAST (6.8 g, 30.7 mmol) and EtOH (129 mg, 2.8 mmol) dropwise. The mixture was heated with microwave heating at 27 °C for 14 h. H 2 O (500 mL) was added and the mixture was extracted with DCM (200 mL x 3), the combined organic layers were concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give 1-bromo-3-(difluoromethyl)-5-iodobenzene (3.2 g, 65% yield) as a solid. 1H NMR (300 MHz, DMSO-d6) δ 8.16 (p, J = 1.2 Hz, 1H), 7.94 (p, J = 1.3 Hz, 1H), 7.81 (p, J = 1.3 Hz, 1H), 7.00 (t, J = 55.3 Hz, 1H).

[00417] Step 2. A mixture of Zn (2.28 g, 34.8 mmol) and I2 (442 mg, 1.74 mmol) in DMF (20 mL) under an Ar atmosphere was stirred at 50 °C for 0.5 h. To this mixture was added a solution of (R)-2-((tert-butoxycarbonyl)amino)-3-iodopropanoate (2.39 g, 7.25 mmol) in DMF (20 mL), and the mixture was stirred at 50 °C for 2 h. After cooling, the mixture was added to 1-bromo-3-(difluoromethyl)-5-iodobenzene (2.90 g, 8.7 mmol), Pd2(dba)3 (239 mg, 0.26 mmol), and tri-2-furylphosphine (162 mg, 0.7 mmol) in DMF (20 mL). The mixture was heated to 70 °C and stirred for 2 h, then H2O (200 mL) was added, and the mixture was extracted with EtOAc (200 mL × 3). The combined organic layers were concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give methyl (S)-3-(3-bromo-5-(difluoromethyl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoate (560 mg, 19% yield) as a solid. 1H NMR (300MHz, DMSO-d6) δ 7.65 (d, J = 10.0Hz, 2H), 7.47 (s, 1H), 7.36 (d, J = 8.4Hz, 1H), 7.00 (t, J = 55.6Hz, 1H), 4.25 (td, J = 9.6, 4.7 Hz, 1H), 3.64 (s, 3H), 3.11 (dd, J = 13.6, 4.9 Hz, 1H), 3.00 - 2.80 (m, 1H), 1.32 (s, 9H).

[00418] Step 3. To a mixture of methyl (S)-3-(3-bromo-5-(difluoromethyl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoate (650 mg, 1.6 mmol) in THF (1.5 mL) at 0 °C under a N2 atmosphere was added LiOH (114 mg, 4.8 mmol) in H2O (1.50 mL). The mixture was stirred at 0 °C for 1 h, then acidified to pH 5 with 1 M HCl. The mixture was extracted with DCM/MeOH (10/1) (100 mL × 3), and the combined organic layers were dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to give (S)-3-(3-bromo-5-(difluoromethyl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid (500 mg), which was used directly in the next step without further purification. LCMS (ESI): m/z [M+H] calculated for C15H18BrF2NO4 393.0; found 392.1.

[00419] Step 4. To a mixture of methyl (3S)-1,2-diazinane-3-carboxylate (475 mg, 3.3 mmol) in DCM (10 mL) at 0 °C under a N2 atmosphere were added N-methylmorpholine (3.34 g, 33.0 mmol) and (S)-3-(3-bromo-5-(difluoromethyl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid (650 mg, 1.7 mmol) and HOBt (45 mg, 0.33 mmol) and EDCI (632 mg, 3.3 mmol). The mixture was warmed to room temperature and stirred for 16 h, then diluted with DCM (100 mL) and H2O. The organic and aqueous layer were separated, and the aqueous layer was extracted with DCM (100 mL × 3). The combined organic layers were dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain methyl (S)-1-((S)-3-(3-bromo-5-(difluoromethyl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate (510 mg, 56% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C21H28BrF2N3O5 519.1; found 520.3.

[00420] Step 5. To a mixture of 4,4,5,5-tetramethyl-2-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (488 mg, 1.92 mmol) and methyl (S)-1-((S)-3-(3-bromo-5-(difluoromethyl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate (500 mg, 0.96 mmol) in 1,4-dioxane (5 mL) was added Pd(dppf)Cl2 (70 mg, 0.07 mmol) and KOAc (236 mg, 2.4 mmol) in portions. The mixture was heated to 90 °C and stirred for 4 h and then diluted with H2O (100 mL). The mixture was extracted with DCM (100 mL × 3), and the combined organic layers were dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain methyl (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(difluoromethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanoyl)hexahydropyridazine-3-carboxylate (423 mg, 73% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C27H40BF2N3O7 567.3; found 568.2.

[00421] Step 6. To a mixture of methyl (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(difluoromethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanoyl)hexahydropyridazine-3-carboxylate (260 mg, was added K2CO3 (163 mg, 1.12 mmol). The mixture was heated to 60 °C and stirred for 16 h, then diluted with H2O (100 mL) and extracted with DCM (100 mL × 3). The combined organic layers were dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain methyl (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(difluoromethyl)-5-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)phenyl)propanoyl)hexahydropyridazine-3-carboxylate (350 mg, 78% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C44H57F2N5O7 805.4; found 806.6.

[00422] Step 7. To a mixture of methyl (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(difluoromethyl)-5-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)phenyl)propanoyl)hexahydropyridazine-3-carboxylate (350 mg, 0.43 mmol) in THF (2.8 mL) at 0 °C was added LiOH H2O (54 mg, 1.3 mmol) in H2O (0.7 mL). The mixture was warmed to room temperature and stirred for 2 h, then acidified to pH 5 with 1 M HCl and extracted with EtOAc (50 mL × 3). The combined organic layers were dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to give (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(difluoromethyl)-5-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)phenyl)propanoyl)hexahydropyridazine-3-carboxylic acid (356 mg) was used directly in the next step without further purification. LCMS (ESI): m/z [M+H] calcd for C43H55F2N5O7 791.4; found 792.6.

[00423] Step 8. To a mixture of (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(difluoromethyl)-5-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)phenyl)propanoyl)hexahydropyridazine-3-carboxylic acid (356 mg, 0.45 mmol) and DIPEA (1.74 g, 13.5 mmol) in DCM were added EDCI (2.41 g, 12.6 mmol) and HOBt (304 mg, 2.3 mmol). The mixture was stirred for 16 h, then H2O was added and the mixture was extracted with EtOAc (200 mL × 3). The combined organic layers were washed with brine (50 mL × 4), dried over anhydrous Na 2 SO 4 , and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give tert-butyl ((63S,4S)-25-(difluoromethyl)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro- 11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (202 mg, 51% yield). LCMS (ESI): m/z [M+H] calcd for C43H53F2N5O6 773.4; found 774.6.

[00424] Step 9. To a mixture of tert-butyl((63S,4S)-25-(difluoromethyl)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (202 mg, 0.26 mmol) in DCM (2 mL) at 0 °C was added TFA (1.0 mL) dropwise. The mixture was stirred at 0 °C for 1.5 h, then concentrated under reduced pressure and azeotropically dried with toluene (3 mL × 3) to give (63S,4S)-4-amino-25-(difluoromethyl)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-5,7-dione, which was used directly in the next without further purification. LCMS (ESI): m/z [M+H] calcd for C38H45F2N5O4 673.3; found 674.5.

[00425] Step 10. To a mixture of (63S,4S)-4-amino-25-(difluoromethyl)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl- 61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-5,7-dione (202 mg, 0.3 mmol) and (2S)-2-[(tert-butoxycarbonyl)(methyl)amino]-3-methylbutanoic acid (139 mg, 0.6 mmol) in THF under an Ar atmosphere was added DIPEA (581 mg, 4.5 mmol), EDCI (86 mg, 0.45 mmol) and HOBt (61 mg, 0.45 mmol) were added. The mixture was stirred for 16 h, then H2O (100 mL) was added and the mixture was extracted with EtOAc (200 mL × 3). The combined organic layers were washed with brine (50 mL × 3), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain tert-butyl ((2S)-1-(((63S,4S)-25-(difluoromethyl)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamate (135 mg, 46% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C49H64F2N6O7 886.5; found 887.6.

[00426] Step 11. To a mixture of tert-butyl ((2S)-1-(((63S,4S)-25-(difluoromethyl)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl) (methyl)carbamate (130 mg, 0.15 mmol) in DCM at 0 °C under a N2 atmosphere was added TFA (1.0 mL) dropwise. The mixture was stirred at 0 °C for 1.5 h, then concentrated under reduced pressure and azeotropically dried with toluene (3 mL × 3) to give (2S)-N-((63S,4S)-25-(difluoromethyl)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)-3-methyl-2-(methylamino)butanamide (130 mg), which was used directly in the next step without further purification. LCMS (ESI): m/z [M+H] calculated for C44H56F2N6O5 786.4; found 787.6.

[00427] Step 12. A mixture of (2S)-N-((63S,4S)-25-(difluoromethyl)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)-3-methyl-2-(methylamino)butanamide (130 mg, 0.17 mmol) and (3S)-1-(prop-2-enoyl)pyrrolidine-3-carboxylic acid (56 mg, 0.33 mmol) in MeCN (1.5 mL) at 0 °C was evaporated to a boiling point. C under a N2 atmosphere were added DIPEA (427 mg, 3.3 mmol) and CIP (69 mg, 0.25 mmol). The mixture was stirred at 0 °C for 1 h, then H2O (100 mL) was added, and the mixture was extracted with EtOAc (200 mL × 3). The combined organic layers were washed with brine (50 mL × 3), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by preparative HPLC to obtain (3S)-1-acryloyl-N-((2S)-1-(((63S,4S)-25-(difluoromethyl)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)-N-methylpyrrolidine-3-carboxamide (58 mg, 36% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C52H65F2N7O7 937.4; found 938.1; 1H NMR (400 MHz, DMSO-d6) δ 8.78 (dd, J = 4.8, 1.7 Hz, 1H), 8.43 - 8.21 (m, 1H), 8.02 (s, 2H), 7.93 - 7.81 (m, 2H), 7.76 (dd, J = 9.3, 3.9 Hz, 1H) 16.8, 4.9, 2.4 Hz, 1H), 5.90 - 5.60 (m, 1H), 5.59 - 5.19 (m, 2H),4.71 (dd, J = 10.7, 3.1 Hz, 1H), 4.40 - 4.17 (m, 3H), 4.12 - 3.90 (m, 3H), 3.85- 3.71 (m, 1H), 3.61 (tdd, J = 23.4, 9.9, 4.3 Hz, 6H), 3.40 - 3.30 (m, 2H), 3.11(d, J = 6.8 Hz, 3H), 3.08 - 2.90 (m, 2H), 2.87 (s, 2H), 2.84 (s, 3H), 2.69 - 2.30 (d, J =16.5 Hz, 1H), 2.30 - 1.79 (m, 5H), 1.75 - 1.45 (m, 2H), 1.40 (d, J = 6.1 Hz, 3H), 1.05 - 0.85 (m, 6H), 0.85 - 0.66 (m, 6H), 0.57 (d, J = 11.8 Hz, 3H). EXAMPLE A741. SYNTHESIS OF (3S)-1 -ACRYLOYL-N-((2S)-1 -(((23S,63S,4S)-11 - ETHYL-12-(2-((S)-1-METHOXYETHYL)PYRIDIN-3-IL)-10,10-DIMETHYL-5,7-DIOXO-61,62,63,64,65,66-HEXA-HYDRO-11H-8-OXA-1(5,3)-INDOLE-6(1,3)-PYRIDAZINE- 2(1,3)-PIPERIDINECYCLOUNDECAPHANE-4-YL)AMINO)-3-METHYL-1 -OXOBUTAN-2-YL) -N-METHYLPYRROLIDINE-3-CARBOXAMIDE

[00428] Step 1. A solution of tert-butyl (3R)-3-(hydroxymethyl)piperidine-1-carboxylate (10 g, 46.45 mmol) in DCM (200 mL) at 0 °C was added PPh3 (15.8 g, 60.4 mmol), Imidazole (4.7 g, 69.7 mmol), and I2 (14.1 g, 55.74 mmol). The reaction suspension was stirred at 20 °C for 17 h, then concentrated under reduced pressure. The residue was purified by silica gel column chromatography to afford tert-butyl (3R)-3-(iodomethyl)piperidine-1-carboxylate (10 g, 66% yield) as an oil. LCMS (ESI): m/z [M+H] calculated for C11H20INO2 325.1; no mass was found.

[00429] Step 2. To a mixture of 3-isopropyl-2,5-dimethoxy-3,6-dihydropyrazine (10.8 g, 58.9 mmol) in THF (150 mL) at -60 °C under a N 2 atmosphere was added n-BuLi (47 mL, 2.5 M in hexane, 117.7 mmol) dropwise. The mixture was warmed to 0 °C and stirred for 2 h, then cooled again to -60 °C and a solution of tert-butyl (3R)-3-(iodomethyl)piperidin-1-yl formate (9.60 g, 29.4 mmol) in THF (50 mL) was added slowly dropwise. The mixture was stirred at -60 °C for 2 h, then warmed to room temperature and stirred for 2 h. Saturated NH4Cl (150 mL) was added slowly and the mixture was extracted with EtOAc (150 mL × 2). The combined organic layers were washed with brine (200 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was reduced under reduced pressure, and the residue was purified by silica gel column chromatography to obtain tert-butyl (3S)-3-{[(2S)-5-isopropyl-3,6-dimethoxy-2,5-dihydropyrazin-2-yl]methyl}piperidin-1-yl formate (5.3 g, 46% yield) as a gum. LCMS (ESI): m/z [M+H] calcd for C20H35N3O4 381.5; found 382.3.

[00430] Step 3. A mixture of tert-butyl(3S)-3-{[(2S)-5-isopropyl-3,6-dimethoxy-2,5-dihydropyrazin-2-yl]methyl}piperidin-1-yl formate (5.30 g, 13.9 mol) in MeCN (4 mL) was added 1M HCl (27.7 mL, 27.7 mmol) dropwise. The mixture was stirred for 2 h, then saturated NaHCO3 was added to ~pH 7-8, then extracted with DCM (30 mL x 2). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give (S)-tert-butyl 3-((S)-2-amino-3-methoxy-3-oxopropyl)piperidine-1-carboxylate (4.3 g, 95% yield) as an oil, which was used in the next step without further purification. LCMS (ESI): m/z [M+H] calcd for C14H26N2O4 286.2; found 287.3.

[00431] Step 4. To a mixture of methyl (S)-tert-butyl 3-((S)-2-amino-3-methoxy-3-oxopropyl)piperidine-1-carboxylate (4.30 g, 15.0 mmol) in EtOAc (30 mL) and H2O (20 mL) at -10 °C was added NaHCO3 (3.77 g, 44.88 mmol). The mixture was stirred at -10 °C for 10 min, then a solution of benzyl chloroformate (3.83 g, 22.44 mmol) was added dropwise. The mixture was warmed to 0 °C and stirred for 1 h, then H2O (50 mL) was added and the mixture was extracted with EtOAc (50 mL × 2). The combined organic layers were dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to give tert-nutyl (3S)-3-[(2S)-2-{[(benzyloxy)carbonyl]amino}-3-methoxy-3-oxopropyl]piperidine-1-carboxylate (4.0 g, 60% yield) as a gum. LCMS (ESI): m/z [M-Boc+H] calculated for C17H24N2O4 320.2; found 321.3.

[00432] Step 5. To a mixture of tert-butyl (3S)-3-[(2S)-2-{[(benzyloxy)carbonyl]amino}-3-methoxy-3-oxopropyl]piperidine-1-carboxylate (1.0 g, 2.38 mmol) in EtOAc (8 mL) was added 2M HCl in EtOAc (11.9 mL, 23.8 mmol). The mixture was stirred for 2 h, then saturated NaHCO3 added until ~pH 8-9 and the mixture was extracted with DCM (30 mL x 3). The combined organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give (2S)-2{[(benzyloxy)carbonyl]amino}-3-[(3S)-piperidin-3-yl]propanoate (740 mg, 91% yield) as a gum. LCMS (ESI): m/z [M+H] calculated for C17H24N2O4 320.2; found 321.2.

[00433] Step 6. To a mixture of (3-{3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl}-1-ethyl-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}indol-5-yl)boranediol (5.47 g, 8.43 mmol) and methyl (2S)-2-{[(benzyloxy)carbonyl]amino}-3-[(3S)-piperidin-3-yl]propanoate (2.70 g, 8.43 mmol) in DCM (70 mL) was added Cu(OAc)2 (6.06 g, 16.86 mmol) and pyridine (2.0 g, 25.3 mmol). The mixture was stirred under an O2 atmosphere for 48 h, then diluted with DCM (200 mL) and washed with H2O (150 mL × 2). The organic layer was dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give methyl 2-{[(benzyloxy)carbonyl]amino}-3-[(3S)-1-(3-{3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl}-1-ethyl-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}indol-5-yl)piperidin-3-yl]propanoate (3.6 g, 42% yield) as a solid. LCMS (ESI): m/z [M/2+H] calculated for C56H70N4O6Si 462.3; found 462.3.

[00434] Step 7. To a mixture of methyl 2-{[(benzyloxy)carbonyl]amino}-3-[(3S)-1-(3-{3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl}-1-ethyl-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}indol-5-yl)piperidin-3-yl]propanoate (3.60 g, 3.57 mmol) in THF (60 mL) and H2O (30 mL) was added LiOH (342 mg, 14.28 mmol). The mixture was stirred for 2 h, then diluted with H2O (150 mL), then 1M HCl was slowly added until ~pH 3-4, and the mixture was extracted with EtOAc (200 mL × 3). The combined organic layers were dried over anhydrous Na2SO4, filtered, and the filtrate concentrated under reduced pressure to obtain 2-{[(benzyloxy)carbonyl]amino}3-[(3S)-1-(3-{3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl}-1-ethyl-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}indol-5-yl)piperidin-3-yl]propanoic acid (3.3 g, 85% yield) as a solid, which was used directly in the next step without further purification. LCMS (ESI): m/z [M/2+H] calcd for C55H68N4O6Si 455.3; found 455.3.

[00435] Step 8. To a mixture of methyl 2-{[(benzyloxy)carbonyl]amino}-3-[(3S)-1-(3-{3-[(tert-butyldiphenylsilyl)oxy]-2,2- dimethylpropyl}-1-ethyl-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}indol-5-yl)piperidin-3-yl]propanoate (3.30 g, 2.91 mmol) in DMF (40 mL) was added methyl (3S)-1,2-diazinan-3-carboxylate (0.42 g, 2.91 mmol), HATU (2.21 g, 5.82 mmol) and DIPEA (2.26 g, 17.46 mmol). The mixture was stirred for 3 h, then poured into gel-H2O and extracted with EtOAc (120 mL × 2). The combined organic layers were washed with saturated NaHCO3 (150 mL), brine (150 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain methyl (3S)-1-[(2S)-2-{[(benzyloxy)carbonyl]amino}-3-[(3S)-1-(3-{3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl}-1-ethyl-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}indol-5-yl)piperidin-3-yl]propanoyl]-1,2-diazinane-3-carboxylate (2.9 g, 95% yield) as a gum. LCMS (ESI): m/z [M/2+H] calcd for C61H78N6O7Si 518.3; found 518.3.

[00436] Step 9. To a mixture of methyl (3S)-1-[(2S)-2-{[(benzyloxy)carbonyl]amino}-3-[(3S)-1-(3-{3-[(tert-butyldiphenylsilyl)oxy]-2,2- dimethylpropyl}-1-ethyl-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}indol-5-yl)piperidin-3-yl]propanoyl]-1,2-diazinane-3-carboxylate (1.70 g, 1.64 mmol) was added to a mixture of 1M TBAF in THF (19.68 mL, 19.68 mmol) and AcOH (1.18 g, 19.68 mmol). The reaction was heated to 60 °C and stirred for 22 h, then diluted with EtOAc (80 mL) and washed with saturated NaHCO3 (80 mL), H2O (60 mL x 2), and brine (60 mL). The organic layer was dried over anhydrous Na2SO4, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by preparative HPLC to obtain methyl (3S)-1-[(2S)-2-{[(benzyloxy)carbonyl]amino}-3-[(3S)-1-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}indol-5-yl]piperidin-3-yl]propanoyl]-1,2-diazinane-3-carboxylate (1.0 g, 73% yield) as a solid. LCMS (ESI): m/z [M/2+H] calcd for C45H60N6O7 399.2; found 399.4.

[00437] Step 10. Mix together methyl (3S)-1-[(2S)-2-{[(benzyloxy)carbonyl]amino}-3-[(3S)-1-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-{2-[(1 S)-1-methoxyethyl]pyridin-3-yl}indol-5-yl]piperidin-3-yl]propanoyl]-1,2-diazinane-3-carboxylate (1.0 g, 1.1 mmol) in 1,2-dichloroethane (10 mL) was added to Me3SnOH (1.42 g, 7.84 mmol). The mixture was heated to 65 °C and stirred for 10 h, then filtered, and the filtrate was concentrated under reduced pressure to obtain (3S)-1-[(2S)-2-{[(benzyloxy)carbonyl]amino}-3-[(3S)-1-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}indol-5-yl]piperidin-3-yl]propanoyl]-1,2-diazinane-3-carboxylic acid (1.0 g, 99% yield) as a gum. The product was used in the next step without further purification. LCMS (ESI): m/z [M/2+H] calcd for C44H58N6O7 392.2; found 392.3.

[00438] Step 11. To a mixture of acid (3S)-1-[(2S)-2-{[(benzyloxy)carbonyl]amino}-3-[(3S)-1-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-{2-[(1S)-1-methoxyethyl] pyridin-3-yl} indole-5-yl] piperidin-3-yl]propanoyl]-1,2-diazinane-3-carboxylic acid (1.0 g, 1.1 mmol) in DCM (30 ml) at 0°C was added HOBT (1.51 g, 11.2 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide HCl (6.44 g, 33.6 mmol) and DIPEA (5.79 g, 44.8 mmol). The mixture was warmed to room temperature and stirred for 6 h, then diluted with H2O and extracted with EtOAc (100 mL x 3). The combined organic layers were dried over anhydrous Na2SO4, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give benzyl N-[(6S,8S,14S)-22-ethyl-21-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}-18,18-dimethyl-9,15-dioxo-16-oxa-2,10,22,28-tetra-azapentacyclo[18.5.2.1A{2,6}.1A{10,14}.0A{23,27}]nonacosa-1(26),20,23(27),24-tetraen-8-yl]carbamate (340 mg, 36% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C44H56N6O6 383.2; found 383.3.

[00439] Step 12. A mixture of benzyl N-[(6S,8S,14S)-22-ethyl-21-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}-18,18-dimethyl-9,15-dioxo-16-oxa-2,10,22,28-tetra-azapentacycle [18.5.2.1 A{2,6}. 1 A{10,14}.0A{23,27}]nonacosa-1(26),20,23(27),24-tetraen-8-yl]carbamate (250mg, 0.33 mmol), Pd/C (100 mg) and NH4Cl (353 mg, 6.6 mmol) in MeOH (5 mL) was stirred under an H2 atmosphere for 4h. The mixture was filtered through Celite and the filtrate was concentrated under reduced pressure. The residue was dissolved in DCM (30 mL) and washed with saturated NaHCO3 (20 mL), H2O (20 mL), and brine (20 mL). The organic layer was dried over Na2SO4, filtered, and the filtrate concentrated under reduced pressure to give (6S,8S,14S)-8-amino-22-ethyl-21-{2-[(1 S)-1-methoxyethyl]pyridin-3-yl}-18,18-dimethyl-16-oxa-2,10,22,28-tetrazapentacyclo[18.5.2.1A{2,6}. 1 A{10,14}.0A{23,27}]nonacosa- 1(26),20,23(27),24-tetraene-9,15-dione which was used in the next step without further purification. LCMS (ESI): m/z [M+H] calculated for C36H50N6O4 631.4; found 631.4.

[00440] Step 13. To a mixture of (6S,8S,14S)-8-amino-22-ethyl-21-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}-18,18-dimethyl-16-oxa- 2,10,22,28-tetra-azapentacyclo[18.5.2.1A{2,6}.1A{10,14}.0A{23,27}]nonacosa-1(26),20,23(27),24-tetraene-9,15-dione (300 mg, 0.48 mmol), acid (2S)-3- methyl-2-{N-methyl-1-[(3S)-1-(prop-2-enoyl)pyrrolidin-3-yl]formamido}butanoic acid (136 mg, 0.48 mmol) and DIPEA (620 mg, 4.8 mmol) in DMF (5 mL) at 0 C was added HATU (183 mg, 0.48 mmol). The mixture was stirred at 0-5 °C for 1 h, then diluted with EtOAc (50 mL), washed with H2O (50 mL × 2), brine (50 mL), dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give (2S)-N-[(6S,8S,14S)-22-ethyl-21-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}-18,18-dimethyl-9,15-dioxo-16-oxa-2,10,22,28-tetra-azepentacyclo[18.5.2.1A{2,6}.1A{10,14}.0A{23,27}]nonacosa-1(26),20,23(27),24-tetraen-8-yl]-3-methyl-2-{N-methyl-1-[(3S)-1-(prop-2-enoyl)pyrrolidine-3-yl]formamido}butanamide (90 mg, 20% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C50H70N8O7 895.5; found 895.4; 1H NMR (400MHz, CD3OD) Hz, 1H), 7.22 (d, J = 12.1 Hz, 1H), 7.09 (dd, J = 8.9, 1.9 Hz, 1H), 6.59 (dt, J = 16.9, 9.9 Hz, 1H), 6.26 (ddd, J = 16.8, 5.0, 1.9 Hz, 1H), 5.80 - 5.67 (m, 1H), 5.59 - 5.46 (m, 1H), 4.93 (d, J = 12.4 Hz, 1H), 4.66 (dd, J = 11.1, 6.4 Hz, 1H), 4.45 (d, J = 12.6 Hz, 1H), 4.28 - 4.19 (m, 1H), 4.13 (dd, J = 14.5, 7.2 Hz, 1H), 4.02 - 3.87 (m, 1H), 3.87 - 3.36 (m, 11H), 3.16 (s, 2H), 3.10 (d, J = 3.4 Hz, 2H), 2.76 (dd, J = 26.9, 13.5 Hz, 3H), 2.61 (s, 1H), 2.35 - 1.97 (m, 5H), 1.78 (dd, J=25.4, 22.1 Hz, 10H), 1.45 (d, J=6.2 Hz, 3H), 1.04 (d, J=6.2 Hz, 3H), 0.95 (dd, J=6.5, 1.8 Hz, 3H), 0.83 (d, J = 6.6 Hz, 3H), 0.72 (d, J = 31.8 Hz, 6H).EXAMPLE A715. SYNTHESIS OF BENZYL ((23S,63S,4S)-11 -ETHYL-12-(2-((S)-1 - METHOXYETHYL)PYRIDIN-3-IL)-10,10-DIMETHYL-5,7-DIOXO-61,62,63,64,65,66-HEXA-HYDRO- 11H-8-OXA-1(5,3)-INDOLE-6(1,3)-PYIDAZINE-2(1,3)PIPERIDINECYCLOUNDECAPHANE- 4-YL)CARBAMATE

[00441] To a solution of ((23S,63S,4S)-4-amino-11-ethyl-12- (2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8- oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-piperidineccloundecaphane-5,7-dione (50 mg, 0.08 mmol), (R)-2-(((1-(4-(dimethylamino)-4-methylpent-2-inoyl)azetidin-3-yl)oxy)methyl)-3-methylbutanoic acid (26 mg, 0.08 mmol) and DIPEA (31 mg, 0.24 mmol) in DMF (1 mL) at 0 C, HATU (30 mg, 0.08 mmol) was added. The reaction mixture was stirred at 0-5 °C for 1 h, then diluted with EtOAc (20 mL), washed with H2O (20 mL × 2) and brine (20 mL). The organic phase was separated and dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give (2R)-2-(((1-(4-(dimethylamino)-4-methylpent-2-ynoyl)azetidin-3-yl)oxy)methyl)-N-((23S,63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10- dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-piperidineclondecaphane-4-yl)-3-methylbutanamide as a solid. 1H NMR (400 MHz, CD3OD) 4.8 Hz, 1H), 7.39 (dd, J = 8.9, 3.1 Hz, 1H), 7.26 (d, J = 16.6 Hz, 1H), 7.11 (d, J = 8.8 Hz, 1H), 5.61 (s, 1H), 4.50 - 4.28 (m, 3H), 4.27 - 4.07 (m, 3H), 3.98 (ddd, J = 25.6, 13.4, 5.1 Hz, 2H), 3.84 - 3.72 (m, 2H), 3.62 (dd, J = 10.7, 4.8 Hz, 2H), 3.55 (d, J = 7.1 Hz, 2H), 3.47 (d, J = 6.5 Hz, 2H), 3.16 (s, 3H), 3.03 - 2.91 (m, 1H), 2.76 (dd,J = 28.7, 15.2 Hz, 3H), 2.62 (s, 1H), 2.40 (t,J = 7.0 Hz, 3H), 2.33 (dd,J = 14.3, 5.0Hz, 4H), 2.05 (d, J= 11.6 Hz, 1H), 1.99 - 1.64 (m, 10H), 1.64 - 1.55 (m, 1H), 1.51 - 1.42 (m, 6H), 1.37 (d, J= 12.3 Hz, 3H), 1.05 (s, 3H), 0.94 (ddd, J= 9.3, 6.7, 2.0 Hz, 6H), 0.76 (D, J = 3.8 Hz, 3H), 0.69 (S, 3H). LCMS (ESI): m/z [M+H] calculated for C53H76N8O7 936.6; found 937.4.EXAMPLE A347. SYNTHESIS OF(2S)-N-[(7S,13S)-21-ETHYL-20-{2-[(1S)-1-METHOXYETHYL] PYRIDINE-3-YL}-17,17-DIMETHYL-8,14-DIOXO-15-OXA-4-TIA-9,21,25,27,28- PENTAZAPENTACYCLE [17.5.2.1A{2,5}.1A{9,13}.0A{22,26}]OCTACOSA-1(25),2,5(28),19,22(26),23-HEXAEN-7-I L]-3-METHYL-2-{N-METHYL-1-[(3S)-1-(PROP-2-ENOYL)PIRROLIDIN-3-YL]FORMAMIDO}BUTANAMIDE

[00442] Step 1. To a mixture of (5-chloro-1H-pyrrolo[3,2- b ]pyridin-3-yl)methanol (3.5 g, 19 mmol), and ((1-methoxy-2-methylprop-1-en-1-yl)oxy)trimethylsilane (6.7 g, 38 mmol) in THF (50 mL) at 0 °C was added TMSOTf (3.8 g, 17 mmol) dropwise. The mixture was stirred at 0-5 °C for 2 h, then diluted with EtOAc (100 mL) and washed with saturated NaHCO3 (50 mL) and brine (50 mL × 2). The organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give methyl 3-(5-chloro-1H-pyrrolo[3,2-b]pyridin-3-yl)-2,2-dimethylpropanoate (3.0 g, 59% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C13H15ClN2O2 266.1; found 267.1.

[00443] Step 2. To a mixture of methyl 3-(5-chloro-1H-pyrrolo[3,2-b]pyridin-3-yl)-2,2-dimethylpropanoate (3.0 g, 11 mmol) in anhydrous THF (50 mL) at 0 °C was added AgOTf (4.3 g, 17 mmol) and I2 (2.9 g, 11 mmol). The mixture was stirred at 0 °C for 2 h, then saturated Na2SO3 (20 mL) and EtOAc (50 mL) were added. The mixture was filtered and the filtrate was washed with brine (50 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give methyl 3-(5-chloro-2-iodo-1H-pyrrolo[3,2-b]pyridin-3-yl)-2,2-dimethylpropanoate (2.3 g, 52% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C13H15ClIN2O2 392.0; found 393.0.

[00444] Step 3. To a mixture of methyl 3-(5-chloro-2-iodo-1H-pyrrolo[3,2-b]pyridin-3-yl)-2,2-dimethylpropanoate (2.3 g, 5.9 mmol) and 2-(2-(2-methoxyethyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.6 g, 7.1 mmol) and K2CO3 (2.4 g, 18 mol) in 1,4-dioxane (25 mL) and H2O (5 mL) under a N2 atmosphere was added Pd(dppf)Cl2.DCM (480 mg, 0.59 mmol). The mixture was heated at 70 °C for 4 h, then diluted with EtOAc (200 mL) and washed with brine (25 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give methyl (S)-3-(5-chloro-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-pyrrolo[3,2-b]pyridin-3-yl-2,2-dimethylpropanoate (2.0 g, 84% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C21H24ClN3O3 401.2; found 402.2.

[00445] Step 4. A mixture of ethyl 3-(5-chloro-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-pyrrolo[3,2-b]pyridin-3-yl)-2-methylpropanoate (2.0 g, 5.0 mmol), Cs2CO3 (3.3 g, 10 mmol), and Etl (1.6 g, 10 mmol) in DMF (30 mL) was stirred for 10 h. The mixture was diluted with EtOAc (100 mL) and washed with brine (20 mL × 4), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain two diastereomers of methyl (S)-3-(5-chloro-1-ethyl-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-pyrrolo[3,2-b]pyridin-3-yl)-2,2-dimethylpropanoate (0.7 g, 32% yield; 0.6 g, 28% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C23H28ClN3O3 429.2; found 430.2.

[00446] Step 5. To a mixture of (S)-3-(5-chloro-1-ethyl-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-pyrrolo[3,2-b]pyridin-3-yl)-2,2-dimethylpropanoate (1.9 g, 4.4 mmol) in anhydrous THF (20 mL) at 0 °C was added LiBH4 (200 mg, 8.8 mmol). The mixture was heated to 60 °C and stirred for 4 h, then saturated NH4Cl (20 mL) and EtOAc (50 mL) added. The aqueous and organic layers were separated, and the organic layer was washed with brine (30 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain (S)-3-(5-chloro-1-ethyl-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-pyrrolo[3,2-b]pyridin-3-yl)-2,2-dimethylpropan-1-ol (1.5 g, 85% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C22H28ClN3O2 401.2; found 402.2.

[00447] Step 6. To a mixture of acid (S)-3-(5-chloro-1-ethyl-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-pyrrolo[3,2-b]pyridin-3-yl)-2,2-dimethylpropan-1-ol (550 mg, 1.37 mmol), (S)-(2-(2-((tert-butoxycarbonyl)amino)-3-methoxy-3-oxopropyl)thiazol-4-yl)boronic acid (907.4 mg, 2.74 mmol, 2 eq) and K2CO3 (568 mg, 4.11 mmol) in 1,4-dioxane (25 mL) and H2O (5 mL) under an N2 atmosphere were added Pd(dppf)Cl2.DCM (89 mg, 0.14 mmol). The mixture was heated to 70 °C and stirred for 4 h, then H2O (50 mL) was added, and the mixture was extracted with EtOAc (100 mL × 3). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain methyl (S)-2-((tert--butycarbonyl)amino)-3-(4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-pyrrylo[3,2-b]pyridin-5-yl)thiazol-2-yl)propanoate d (440 mg, 22% yield) as a solid, which was used directly in the next step. LCMS (ESI): m/z [M+H] calculated for C34H45N5O6S 651.3; found 652.3.

[00448] Step 7. To a mixture of (2S)-methyl 2-((tert-butoxycarbonyl)amino)-3-(4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-pyrrolo[3,2-b]pyridin-5-yl)thiazol-2-yl)propanoate (280 mg, 0.43 mmol) in MeOH (4 mL) was added a solution of LiOH (51 mg, 2.2 mmol) in H2O (2 mL). The mixture was stirred for 5 h, then the pH was adjusted to ~3-4 by addition of 1 M HCl. The mixture was diluted with H2O (30 mL) and extracted with EtOAc (15 mL × 3). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to give (2S)-2-((tert-butoxycarbonyl)amino)-3-(4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-pyrrolo[3,2-b]pyridin-5-yl)thiazol-2-yl)propanoic acid (280 mg) as a solid, which was used directly in the next step without further purification. LCMS (ESI): m/z [M+H] calcd for C33H43N5O6S 637.3; found 638.3.

[00449] Step 8. To a mixture of (2S)-2-((tert-butoxycarbonyl)amino)-3-(4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-pyrrolo[3,2-b]pyridin-5-yl)thiazol-2-yl)propanoic acid (274 mg, 0.43 mmol) and methyl (3S)-1,2-diazinane-3-carboxylate (280 mg, 0.64 mmol) in DMF (3 mL) at 0-5 °C was added a solution of HATU (245 mg, 0.64 mmol) and DIPEA (555 mg, 4.3 mmol) in DMF (2 mL). The mixture was stirred for 1 h, then diluted with EtOAc (20 mL) and H2O (20 mL). The aqueous and organic layers were partitioned, and the organic layer was washed with H2O (20 mL × 3), brine (20 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain methyl (3S)-1-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-3-{4-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}pyrrolo[3,2-b]pyridin-5-yl]-1,3-thiazol-2-yl}propanoyl]-1,2-diazinane-3-carboxylate (230 mg, 70% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C39H53N7O7S 763.4; found 764.3.

[00450] Step 9. To a mixture of methyl (3S)-1-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-3-{4-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-{2-[(1 S)-1-methoxyethyl]pyridin-3-yl}pyrrolo[3,2-b]pyridin-5-yl]-1,3-thiazol-2-yl}propanoyl]-1,2-diazinane-3-carboxylate (230 mg, 0.3 mmol) in DCE (3 mL) under a N2 atmosphere was added Me3SnOH (300 mg). The mixture was heated to 65 °C and stirred for 16 h, then concentrated under reduced pressure. The residue was diluted with EtOAc (20 mL), washed with H2O (20 mL) and brine (10 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain (3S)-1-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-3-{4-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}pyrrolo[3,2-b]pyridin-5-yl]-1,3-thiazol-2-yl}propanoyl]-1,2-diazinane-3-carboxylic acid (200 mg) as foam, which was used directly in the next step without further purification. LCMS (ESI): m/z [M+H] calcd for C38H51N7O7S 749.4; found 750.3.

[00451] Step 10. To a mixture of acid (3S)-1-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-3-{4-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-{2-[(1 S)-1-methoxyethyl]pyridin-3-yl}pyrrolo[3,2-b]pyridin-5-yl]-1,3-thiazol-2-yl}propanoyl]-1,2-diazinan-3-carboxylic acid (245 mg, 0.32 mmol) in DCM (50 mL) at 0-5 °C were added HOBT (432 mg, 3.2 mmol), EDCI HCl (1.8 g, 9.6 mmol) and DIPEA (1.65 g, 12.8 mmol). The mixture was warmed to room temperature and stirred for 16 h, then concentrated under reduced pressure. The residue was diluted with EtOAc (20 mL) and H2O (20 mL), and the aqueous and organic layers were partitioned. The organic layer was washed with H2O (30 mL × 3), brine (30 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative TLC to obtain tert-butyl((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-thiazole-1(5,3)-pyrrolo[3,2-b]pyridine-6(1,3)-pyridazinecycloundecaphane-4-yl)carbamate (100 mg, 43% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C38H49N7O6S 731.4; found 732.3.

[00452] Step 11. A mixture of tert-butyl ((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo- 61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-thiazole-1(5,3)-pyrrolo[3,2-b]pyridine-6(1,3)-pyridazinacloundecaphane-4-yl)carbamate (80 mg, 0.11 mmol) in DCM (0.6 mL) and TFA (0.2 mL) was stirred for 1 h. The mixture was concentrated under reduced pressure to give (63S,4S,Z)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-thiazole-1(5,3)-pyrrolo[3,2-b]pyridin-6(1,3)-pyridazinaccloundecaphane-5,7-dione (72 mg, 95% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C33H41N7O4S 631.3; found 632.3.

[00453] Step 12. To a mixture of (63S,4S,Z)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66- hexahydro-11H-8-oxa-2(4,2)-thiazole-1(5,3)-pyrrolo[3,2-b]pyridine-6(1,3)-pyridazinecycloundecaphane-5,7-dione(120 mg, 0.39 mmol) and DIPEA (335 mg, 2.6 mmol) in DMF (1 mL) at 0 °C was added HATU (60 mg, 0.16 mmol). The mixture was stirred at 0 °C for 1 h, then diluted with H2O (110 mL) and extracted with EtOAc (80 mL × 2). The combined organic layers were washed with H2O (100 mL), brine (100 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative TLC to give (3S)-1-acryloyl-N-((2S)-1-(((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-thiazole-1(5,3)-pyrrolo[3,2-b]pyridine-6(1,3)-pyridazinecycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)-N-methylpyrrolidine-3-carboxamide (1.8 mg, 2% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C47H61N9O7S 895.4; found; and 896.3;1H NMR (400 MHz, CD3OD) δ 8.72 (d, J = 4.5 Hz, 1H), 7.98 - 7.77 (m, 3H), 7.72 (dd, J = 12.0, 8.6 Hz, 1H), 7.54 (dd, J = 7.6, 4.8 Hz, 1H) 3.89 - 3.65 (m,6H), 3.50 (m, 4H), 3.34 (d, J = 6.2 Hz, 3H), 3.12 (d, J = 4.0 Hz, 2H), 3.00 (s,1H), 2.73 (m, 1H), 2.48 - 2.37 (m, 1H), 2.31 - 2.07 (m, 4H), 1.88 (d, J = 11.2 Hz, 1H), 1.71 (d, J = 12.8 Hz, 1H), 1.44 (m, 7H), 0.97 (dd, J = 6.2, 4.4 Hz, 3H), 0.92 - 0.84 (m, 8H), 0.41 (d, J = 6.2 Hz, 3H).EXAMPLE 647. SYNTHESIS OF 1-(4-(DIMETHYLAMINO)-4-METHYLPENT-2-INOYL)-N-((2S)-1- (((22S,63S,4S)-11 -ETHYL-12-(2-((S)-1 -METHOXYETHYL)PYRIDIN-3-YL)-1 0,10-DIMETHYL-5,7- DIOXO-61 ,62,63,64,65,66-HEXA-HYDRO-11H-8-OXA-2(4,2)-MORPHOLINE-1(5,3)- INDOLA-6(1,3)-PYRDAZINECYCLOUNDECAPHANE-4-YL)AMINO)-3-METHYL-1-OXOBUTAN-2-YL) -4-FLUORO-N-METHYLPIPERIDINE-4-CARBOXAMIDE

[00454] Step 1. A mixture of 1-[tert-butoxy)carbonyl]-4-fluoropiperidine-4-carboxylic acid (2.0 g, 8.1 mmol) in DCM (20 mL) was added oxalic dichloride (1.34 g, 10.5 mmol) and DMF (30 mg, 0.4 mmol). The resulting solution was stirred at room temperature for 1 h. Et3N (3.2 g, 3.2 mmol) and (2S)-3-methyl-2-(methylamino)butanoic acid (1.25 g, 9.5 mmol) were added, and the mixture was stirred at room temperature for 1 h. H2O (100 mL) was added, and the mixture was extracted with EtOAc (50 mL × 3). The combined organic layers were concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give tert-butyl (S)-4-((1-tert-butoxy)-3-methyl-1-oxybutan-2-yl)(methyl)carbamoyl)-4-fluoropiperidine-1-carboxylate (1.34 g, 45% yield) as a solid. LCMS (ESI): m/z [M+Na] calculated for C21H37FN2O5Na 439.3; found 439.3.

[00455] Step 2. A mixture of tert-butyl (S)-4-((1-(tert-butoxy)-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)-4-fluoropiperidine-1-carboxylate (290 mg, 0.70 mmol) in DCM (4 mL) and TFA (2 mL) was stirred at room temperature for 2 h, then concentrated under reduced pressure to give N-(4-fluoropiperidine-4-carbonyl)-N-methyl-L-valine, which was used directly in the next step without further purification. LCMS (ESI): m/z [M+H] calcd for C12H21FN2O3 260.2; found 261.2.

[00456] Step 3. To a solution of tert-butyl N-(4-fluoropiperidine-4-carbonyl)-N-methyl-L-valinate (1.7 g, 5.3 mmol), sodium 4-(dimethylamino)-4-methylpent-2-ynoate (1.67 g, 9.4 mmol), and Et3N (2.73 g, 36.9 mmol) in DMF (20 mL) stirred at 5 °C was added T3P (4.11 g, 10.7 mmol, 50 wt % in EtOAc). The reaction mixture was stirred at 5 °C for 1 h. The resulting mixture was quenched with H2O (100 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were concentrated and purified by silica gel column chromatography to give tert-butyl N-(1-(4-(dimethylamino)-4-methylpent-2-ynoyl)-4-fluoropiperidine-4-carbonyl)-N-methyl-L-valinate (1.6 g, 74.0% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C24H40FN3O4 453.3; found 454.2.

[00457] Step 4. To a solution of tert-butyl N-(1-(4-(dimethylamino)-4-methylpent-2-ynoyl)-4-fluoropiperidine-4-carbonyl)-N-methyl-L-valinate (50 mg, 0.11 mmol) in DCM (2 mL) was added TFA (1 mL). The reaction mixture was stirred at 20 °C for 2 h, then concentrated under reduced pressure to afford N-(1-(4-(dimethylamino)-4-methylpent-2-ynoyl)-4-fluoropiperidine-4-carbonyl)-N-methyl-L-valine. It was used for the next step directly without further purification. LCMS (ESI): m/z [M+H] calculated for C20H32FN3O4 397.2; found 398.3.

[00458] Step 5. To a solution of tert-butyl (2R)-2-(hydroxymethyl)morpholin-4-yl formate (50 g, 230 mmol) in EtOAc (1 L) was added TEMPO (715 mg, 4.6 mmol) and NaHCO3 (58 g, 690 mmol) at 20 °C. The mixture was cooled to -50 °C, then TCCA (56 g, 241 mmol) in EtOAc (100 mL) was added dropwise over 30 min. The reaction mixture was heated at 5 °C for 2 h, then quenched with 10% Na2S2O3 (200 mL) and stirred for 20 min. The resulting mixture was filtered, and the organic phase was separated from the filtrate. The aqueous phase was extracted with EtOAc (100 mL × 2). The combined organic layers were washed with H2O (100 mL) and brine (100 mL) and dried over anhydrous Na2SO4. The organic layer was concentrated under reduced pressure to afford tert-butyl (2R)-2-formylmorpholin-4-yl formate (50 g, crude) as an oil.

[00459] Step 6. To a solution of tert-butyl (2R)-2-formylmorpholin-4-yl formate (49 g, 153 mmol) and methyl 2-{[(benzyloxy)carbonyl]amino}-2-(dimethoxyphosphoryl)acetate (60 g, 183 mmol) in CAN (300 mL) was added tetramethylguanidine (35 g, 306 mmol) at 0~10 °C. The reaction mixture was stirred at 10 °C for 30 min and then heated at 20 °C for 2 h. The reaction mixture was diluted with DCM (200 mL) and washed with citric acid (10%, 200 mL) and 10% aqueous NaHCO3 solution (200 mL). The organic phase was concentrated under reduced pressure and purified by silica gel column chromatography to afford tert-butyl (S,Z)-2-(2-(((benzyloxy)carbonyl)amino)-3-methoxy-3-oxoprop-1-en-1-yl)morpholine-4-carboxylate (36 g, 90% yield) as a solid. LCMS (ESI): m/z [M+Na] calcd for C21H28N2O4 420.2; found: 443.1

[00460] Step 7. To a solution of tert-butyl (S,Z)-2-(2-(((benzyloxy)carbonyl)amino)-3-methoxy-3-oxoprop-1-en-1-yl)morpholine-4-carboxylate (49 g, 0.12 mol) in MeOH (500 mL) was added (S,S)-Et-DUPHOS-Rh (500 mg, 0.7 mmol). The mixture was stirred at 25 °C under an H2 atmosphere (60 psi) for 48 h. The reaction was concentrated and purified by chromatography to give tert-butyl (S)-2-((S)-2-(((benzyloxy)carbonyl)amino)-3-methoxy-3-oxopropyl)morpholine-4-carboxylate (44 g, 89.8% yield) as a solid. LCMS (ESI): m/z [M+Na] calcd for C21H30N2O7 422.2; found: 445.2.

[00461] Step 8. To a stirred solution of tert-butyl (S)-2-((S)-2-(((benzyloxy)carbonyl)amino)-3-methoxy-3-oxopropyl)morpholine-4-carboxylate (2.2 g, 5.2 mmol) in EtOAc (2 mL) was added HCl/EtOAc (25 mL) at 15 °C. The reaction was stirred at 15 °C for 2 h, then concentrated under reduced pressure to give methyl (S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-morpholin-2-yl)propanolamine (1.51 g, 90.4% yield) as an oil. LCMS (ESI): m/z [M+H] calcd for C16H22N2O5 322.1; found 323.2.

[00462] Step 9. To a solution of 3-(5-bromo-1-ethyl-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}indol-3-yl)-2,2-dimethylpropan-1-ol (100 g, 0.22 mol) and 1H-imidazole (30.6 g, 0.45 mol) in DCM (800 mL) was added TBSCl (50.7 g, 0.34 mol) in DCM (200 mL) at 0 °C. The reaction was stirred at 25 °C for 2 h. The resulting solution was washed with H2O (300 mL × 3) and brine (200 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified with silica gel column chromatography to give (S)-5-bromo-3-(3-((tert-butyldimethylsilyl)oxy)-2,2-dimethylpropyl)-1-ethyl-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-indole (138 g, 90% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C29H43BrN2O2Si 558.2; found 559.2.

[00463] Step 10. To a stirred solution of Intermediate 1 (50 g, 89.3 mmol) in dioxane (500 mL) was added methyl (2S)-2-{[(benzyloxy)carbonyl]amino}-3-[(2S)-morpholin-2-yl]propanoate from step 1 (31.7 g, 98.2 mmol), RuPhos (16.7 g, 35.7 mmol), Di-mu-chlorobis(2-amino-1,1-biphenyl-2-yl-C,N)dipalladium(II) (2.8 g, 4.4 mmol), and cesium carbonate (96 g, 295 mmol) followed by RuPhos-Pd-G2 (3.5 g, 4.4 mmol) at 105 °C under a N2 atmosphere. The reaction mixture was stirred for 6 h at 105 °C under a N2 atmosphere. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC chromatography to give methyl (2S)-2-{[(benzyloxy)carbonyl]amino}-3-[(2S)-4-(3-{3-[(tert-butyldimethylsilyl)oxy]-2,2-dimethylpropyl}-1-ethyl-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}indol-5-yl)morpholin-2-yl]propanoate (55 g, 73% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C45H64N4O7Si 800.5; found 801.5.

[00464] Step 11. To a solution of methyl ((2S)-2-{[(benzyloxy)carbonyl]amino}-3-[(2S)-4-(3-{3-[(tert-butyldimethylsilyl)oxy]-2,2-dimethylpropyl}-1-ethyl-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}indol-5-yl)morpholin-2-yl]propanoate (10 g, 12 mmol) in THF (270 mL) was added LiOH (1.3 g, 31 mmol) in H2O (45 mL) at 20 °C. The reaction was stirred at 20 °C for 2 h, then treated with 1 N HCl to adjust the pH to 4~5 at 0~5 °C. The resulting mixture was extracted with EtOAc (50 mL × 2). The layers Combined organic layers were washed with brine and dried over anhydrous Na2SO4. The organic phase was then concentrated under reduced pressure to give (2S)-2-{[(benzyloxy)carbonyl]amino}-3-[(2S)-4-(3-{3-[(tert-butyldimethylsilyl)oxy]-2,2-dimethylpropyl}-1-ethyl-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}indol-5-yl)morpholin-2-yl]propanoic acid (9.5 g, 97% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C44H62N4O7Si 786.4; found 787.4.

[00465] Step 12. To a stirred solution of (2S)-2-{[(benzyloxy)carbonyl]amino}-3-[(2S)-4-(3-{3-[(tert-butyldimethylsilyl)oxy]-2,2-dimethylpropyl}-1-ethyl-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}indol-5-yl)morpholin-2-yl]propanoic acid (10 g, 12.7 mmol) in DMF (150 mL), was added methyl (S)-hexahydropyridazine-3-carboxylate (2 g, 14 mmol), then cooled to 0 °C, DIPEA (32.8 g, 254 mmol) was added followed by HATU (9.7 g, 25.4 mmol) at 0~5 °C. The reaction mixture was stirred at 0~5 °C for 1 h. The resulting mixture was diluted with EtOAc (500 mL) and H2O (200 mL). The organic layer was separated and washed with H2O (100 mL × 2) and brine (100 mL), dried over anhydrous sodium sulfate. The solution was filtered and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give methyl (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-4-(3-(3-((tert-butyldimethylsilyl)oxy)-2,2-dimethylpropyl)-1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)morpholin-2-yl)propanoyl)hexahydropyridazine-3-carboxylate (8 g, 70% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C50H72N6O8Si 912.5; found 913.4.

[00466] Step 13. A solution of methyl (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-4-(3-(3-((tert-butyldimethylsilyl)oxy)-2,2-dimethylpropyl)-1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)morpholin-2-yl)propanoyl)hexahydropyridazine-3-carboxylate (8.5 g, 9 mmol) in THF (8 mL) was added to a mixture of tetrabutylammonium fluoride (1 M in THF, 180 mL, 180 mmol) and AcOH (11 g, 200 mmol) at 20 °C. The reaction mixture was stirred at 75 °C for 3 h. The resulting mixture was diluted with EtOAc (150 mL) and washed with H2O (20 mL × 6). The organic phase was concentrated under reduced pressure to obtain methyl (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)morpholin-2-yl)propanoyl)hexahydropyridazine-3-carboxylate (7.4 g, 100% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C44H58N6O8 799.4; found 798.4.

[00467] Step 14. To a solution of methyl (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)morpholin-2-yl)propanoyl)hexahydropyridazine-3-carboxylate (8 g, 10 mmol) in THF (200 mL) was added lithium hydroxide (600 mg, 25 mmol) in H2O (30 mL). The reaction mixture was stirred at 20 °C for 1 h, then treated with 1 N HCl to adjust the pH to 4~5 at 0~5 °C, and extracted with EtOAc (500 mL × 2). The organic phase was washed with brine and concentrated under reduced pressure to give (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)morpholin-2-yl)propanoyl)hexahydropyridazine-3-carboxylic acid (8 g, crude) as a solid. LCMS (ESI): m/z [M+H] calcd for C43H56N6O8 784.4; found 785.4.

[00468] Step 15. To a stirred solution of (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)morpholin-2-yl)propanoyl)hexahydropyridazine-3-carboxylic acid (8 g, 10.2 mmol) and DIPEA (59 g, 459 mmol) in DCM (800 mL) was added EDCI (88 g, 458 mmol) and HOBT (27.6 g, 204 mmol) at 25 °C under an argon atmosphere. The reaction mixture was stirred at 25 °C for 16 h. The resulting mixture was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give benzyl((22S,63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholine-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)carbamate (5 g, 66% yield) as a solid; LCMS (ESI): m/z [M+H] calcd for C43H54N6O7 766.4; found 767.4.

[00469] Step 16. To a solution of benzyl((22S,63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholine-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)carbamate (400 mg, 0.5 mmol) in MeOH (20 mL) was added Pd/C (200 mg) and ammonium acetate (834 mg, 16 mmol) at 20 °C under an atmosphere of H2 and the mixture was stirred for 2 h. Then, the resulting mixture was filtered and concentrated under reduced pressure. The residue was redissolved in DCM (20 mL) and washed with H2O (5 mL × 2), then concentrated under reduced pressure to afford (22S,63S,4S)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholine-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-5,7-dione (320 mg, 97% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C35H48N6O5 632.4; found 633.3.

[00470] Step 17. To a solution of (22S,63S,4S)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholine-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-5,7-dione (50 mg, 0.079 mmol), N-(1-(4-(dimethylamino)-4-methylpent-2-ynoyl)-4-fluoropiperidine-4-carbonyl)-N-methyl-L-valine (47 mg, 0.12 mmol) in DMF (2 mL) stirred at 0 °C was added HATU (36 mg, 0.12 mmol) 0.09 mmol) and DIPEA (153 mg, 1.2 mmol) dropwise. The reaction was stirred at 0 °C for 1 h. The resulting mixture was purified by reverse phase to yield 1-(4-(dimethylamino)-4-methylpent-2-inoyl)-N-((2S)-1-(((22S,63S,4S)-11-ethyl-12-(2-((S)-1- methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholine-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-4-yl)amino)- 3-methyl-1-oxobutan-2-yl)-4-fluoro-N-methylpiperidine-4-carboxamide (11.9 mg, 13.9% yield) as a solid. 1H NMR (400MHz, CD3OD) δ 8.71(dd, J=4.8, 1.7Hz, 1H), 7.86(d, J=7.8Hz, 1H), 7.51(dd, J=7.8, 4.8Hz, 1H), 7.39(d, J=8.9Hz, 1H), 7.15 - 7.04 (m, 2H), 5.67 (d, J = 8.8 Hz, 1H), 4.62 (d, J = 11.2 Hz, 1H), 4.46 (d, J = 12.4 Hz, 1H), 4.39 - 4.27 (m, 2H), 4.23 (d, J = 6.1 Hz, 1H), 4.17 - 4.08 (m, 1H), 3.93 (s, 2H), 3.86 (s, 1H), 3.84 - 3.76 (m, 2H), 3.74 - 3.65 (m, 2H), 3.63 - 3.51 (m, 2H), 3.27 - 3.23 (m, 1H), 3.22 - 3.11 (m, 6H), 3.0 - 2.89 (m, 2H), 2.85 - 2.75 (m, 2H), 2.74 - 2.55 (m, 2H), 2.36 (d, J = 8.2 Hz, 6H), 2.32 - 2.21 (m, 2H), 2.20 - 2.02 (m, 5H), 1.92 (d, J = 12.5 Hz, 2H), 1.69 (dd, J = 43.8, 12.6 Hz, 2H), 1.46 (dt, J = 8.0, 4.9 Hz, 9H), 1.03 (d, J = 3.5 Hz, 3H), 0.90 (dd, J = 48.3, 6.5 Hz, 6H), 0.77 (d, J = 3.0 Hz, 3H), 0.69 (s, 3H). LCMS (ESI): m/z [M+H] calculated for C55H78FN9O8 1011.6; found 1012.5.EXAMPLE A375. SUMMARY OF (2R)-2-(((1-(4-(DIMETHYLAMINO)-4-METHYLPENT-2-INOYL)AZETIDIN-3-YL)OXY)METHYL)-N -((22S,63S,4S)-12-(2-((S)-1-METHOXYETHYL)PYRIDIN-3-IL)-10,10-DIMETHYL-5,7-DIOXO -11-(2,2,2-TRIFLUOROETHYL)-61,62,63,64,65,66-HEXA-HYDRO-11H-8-OXA-2(4,2)-MORPHOLINE-1(5,3)-INDOLE-6(1,3)-PYRDAZINECYCLOUNDECAPHANE-4-YL)-3-METHYLBUTANAMIDE

[00471] Step 1. To a mixture of 5-bromo-3-{3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl}-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}-1H-indole (10.0 g, 15.2 mmol) in anhydrous DMF (120 mL) at 0 °C under a N2 atmosphere was added NaH, 60% dispersion in oil (1.2 g, 30.4 mmol), and 2,2,2-trifluoroethyl trifluoromethanesulfonate (35.4 g, 152 mmol). The mixture was stirred at 0 °C for 1 h, then saturated NH4Cl (30 mL) was added and the mixture was extracted with EtOAc (100 mL × 3). The combined organic layers were washed with brine, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain (S)-5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indole (8 g) as an oil and the other atropisomer (6 g, 48% yield) as an oil. LCMS (ESI): m/z [M+H] calcd for C39H44BrF3N2O2Si 736.2; found 737.1.

[00472] Step 2. To a mixture of (S)-5-bromo-3-(3-((tert-methoxyethyl)oxy)-2,2-dimethylpropyl)-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indole (7.2 g, 9.7 mmol) in toluene (80 mL) under An N2 atmosphere was added 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (2.7 g, 10.6 mmol), KOAc (1.9 g, 19.4 mmol) and Pd(dppf)Cl2 DCM (0.8 g, 0.1 mmol). The mixture was heated to 90 °C and stirred for 8 h, then saturated NH 4 Cl (30 mL) was added, and the mixture was extracted with EtOAc (40 mL × 3). The combined organic layers were dried over anhydrous Na 2 SO 4 and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain (S)-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-(1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2-trifluoroethyl)-1H-indole (6.1 g, 64% yield) as an oil. LCMS (ESI): m/z [M+H] calculated for C45H56BF3N2O4Si 784.4; found 785.3.

[00473] Step 3. To a mixture of (S)-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-(1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2-trifluoroethyl)-1H-indole (33 g, 42 mmol) in THF (120 mL) and MeOH (330 mL) at 0 °C under a N2 atmosphere was added MeB(OH)2 (50.4 g, 841 mmol) then a mixture of NaOH (33.6 g, 841 mmol) in H2O (120 mL). The mixture was warmed to room temperature and stirred for 16 h, then concentrated under reduced pressure. H2O (500 mL) was added to the residue, and the mixture was extracted with EtOAc (300 mL × 3). The combined organic layers were washed with brine (300 mL), H2O (300 mL), then concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give (S)-(3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)1H-indol-5-yl)boronic acid (20 g, 68% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C39H46BF3N2O4Si 702.3; found 703.3.

[00474] Step 4. Note: Three reactions were run in parallel - yield reflects the sum of the products.

[00475] A mixture of (S)-(3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)boronic acid (1.85 g, 5.6 mmol) and methyl (S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-morpholin-2-yl)propanoate in DCM (150 mL) under air was added pyridine (1.35 g, 16.9 mmol) and Cu(OAc)2 (2.0 g, 11.3 mmol). The mixture was stirred for 48 h, then concentrated under reduced pressure. H2O (300 mL) was added to the residue, and the mixture was extracted with EtOAc (300 mL × 2). The combined organic layers were washed with brine (100 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column to obtain methyl (S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-4-(3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)morpholin-2-yl)propanoate (9.2 g, 55% yield) as a solid. LCMS (ESI): m/z [M/2+H] calculated for C55H65F3N4O7Si 490.2; found 490.3.

[00476] Step 5. To a mixture of methyl (S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-4-(3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)morpholin-2-yl)propanoate (10.8 g, 11.0 mmol) in THF (50 mL) was added LiOH (528 mg, 22 mmol) in H2O (10 mL). The mixture was stirred for 1 h, then cooled to 0-5°C and acidified to pH~7 using 2N HCl (10 mL). The mixture was extracted with DCM (100 mL × 2), and the combined organic layers were dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to obtain (S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-4-(3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)morpholin-2-yl)propanoic acid (10.6 g, 100% yield) as a solid. LCMS (ESI): m/z [M/2+H] calcd for C54H63F3N4O7Si 483.2; found 483.3.

[00477] Step 6. To a mixture of (S)-2-(((benzyloxy)carbonyl)amino acid)-3-((S)-4-(3-(3-((tert-butyldiphenylsilyl)oxy)-2,2- dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)morpholin-2-yl)propanoic acid (10.6 g, 11.0 mmol) and methyl (3S)-1,2-diazinan-3-carboxylate (15.8 g, 22.0 mmol) in DMF (150 mL) at 0°C was added DIPEA (28.4 g, 220 mmol) and HATU (8.4 g, 22.0 mmol). The mixture was stirred at 0~5 °C for 1 h, then EtOAc (500 mL) was added and the mixture was washed with H2O (200 mL × 2), brine (100 mL), dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain methyl (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-4-(3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)morpholin-2-yl)propanoyl)hexahydropyridazine-3-carboxylate (11 g, 90% yield) as a solid. LCMS (ESI): m/z [M/2+H] calcd for C60H73F3N6O8Si 546.3; found 546.3.

[00478] Step 7. To a mixture of (S)-1-((S)-2-(((benzyloxy)carbonyl) amino)-3-((S)-4-(3- (3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)morpholin-2-yl)propanoyl)hexahydropyridazine-3-carboxylate (11.0 g, 10.1 mmol) in THF (10 mL) was added to a mixture of AcOH (21.2 g, 353 mmol) and 1M TBAF in THF (300 mL, 300 mmol). The mixture was heated to 80 °C and stirred for 16 h, then concentrated under reduced pressure. EtOAc (800 mL) was added to the residue, and the mixture was washed with H2O (80 mL × 6), concentrated under reduced pressure, and the residue was purified by preparative HPLC to obtain methyl (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-4-(3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)morpholin-2-yl)propanoyl)hexahydropyridazine-3-carboxylate (7.9 g, 91% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C44H55F3N6O8 852.4; found 853.3.

[00479] Step 8. To a mixture of methyl (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-4-(3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)morpholin-2-yl)propanoyl)hexahydropyridazine-3-carboxylate (7.9 g, 9.3 mmol) in THF (50 mL) was added LiOH (443 mg, 18.5 mmol) in H2O (10 mL). The mixture was stirred for 1 h, then cooled to 0-5 °C and acidified to ~pH 7 with 2N HCl (9 mL). The mixture was extracted with DCM (100 mL × 2), and the combined organic layers were dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to obtain (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-4-(3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)morpholin-2-yl)propanoyl)hexahydropyridazine-3-carboxylic acid (7.6 g, 98% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C43H53F3N6O8 838.4; found 839.3.

[00480] Step 9. To a mixture of (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-4-(3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)morpholin-2-yl)propanoyl)hexahydropyridazine-3-carboxylic acid (7.6 g, 9.0 mmol) and DIPEA (52.3 g, 405 mmol) in DCM (800 mL) under an Ar atmosphere was added EDCI (77.6 g, 405 mmol) and HOBT (12 g, 90 mmol). The mixture was stirred for 16 h, then concentrated under reduced pressure. The residue was diluted with EtOAc (500 mL), washed with H2O (100 mL × 2), and filtered. The organic layer was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give benzyl((22S,63S,4S)-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholine-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)carbamate (6.1 g, 74% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C43H51F3N6O7 820.4; found 821.3.

[00481] Step 10. To a mixture of benzyl ((22S,63S,4S)-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2- trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholine-1(5,3)-indole-6(1,3)-pyridazinecycloundephane-4-yl)carbamate (700 mg, 0.85 mmol) in MeOH (30 mL) was added 10% Pd in C (317 mg) and NH4Cl (909 mg). The mixture was stirred under an atmosphere of H2 (1 atm) for 16 h, then filtered through Celite, and the filter cake was washed with MeOH (150 mL). The filtrate was concentrated under reduced pressure, DCM (20 mL) was added to the residue, and the mixture was washed with saturated NaHCO3 (20 mL × 3). The organic layer was concentrated under reduced pressure to obtain (22S,63S,4S)-4-amino-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholine-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-5,7-dione (660 mg, 95% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C35H45F3N6O5 686.3; found 687.3; 1H NMR (400 MHz, CDCl3) δ 8.80 (dd, J = 4.7, 1.7 Hz, 1H), 7.66 (d, J = 7.4 Hz, 1H), 7.43 - 7.30 (m, 2H), 7.12 - 7.01 (m, 2H), 4.90 - 4.83 (m, 1H), 4.68 (d, J = 12.5 Hz, 1H), 4.57 (dd, J = 16.2, 8.1 Hz, 1H), 4.24 (q, J = 6.1 Hz, 1H), 4.08 (d, J = 10.6 Hz, 1H), 3.97 - 3.82 (m, 4H), 3.80 - 3.68 (m, 2H) 1H), 2.19 - 2.04 (m, 4H), 1.96 (d, J = 13.6 Hz, 2H), 1.80 - 1.71 (m, 2H), 1.66 - 1.59 (m, 1H), 1.47 (d, J = 6.1 Hz, 3H), 0.88 (s, 3H), 0.42 (s, 3H).

[00482] Step 11. To a mixture of (22S,63S,4S)-4-amino-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-11-(2,2,2-trifluoroethyl)- 61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholine-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-5,7-dione (300 mg, 0.4 mmol), acid (2R)-2-[({1- [4-(dimethylamino)-4-methylpent-2-inoyl]azetidin-3-yl}oxy)methyl]-3-methylbutanoic acid (157 mg, 0.48 mmol) and DIPEA (569.0 mg, 0.4 mmol) in DMF (5 mL) at 0 C was added HATU (217 mg, 0.57 mmol). The mixture was stirred at 0 °C for 0.5 h, then diluted with H2O and extracted with EtOAc (20 mL × 3). The combined organic layers were washed with brine (20 mL × 3), dried over Na2SO4 , and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by preparative TLC to give (2R)-2-(((1-(4-(dimethylamino)-4-methylpent-2-ynoyl)azetidin-3-yl)oxy)methyl)-N-((22S,63S,4S)-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholine-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)-3-methylbutanamide (200 mg, 46% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C52H71F3N8O8 992.5; found 993.4; 1H NMR (400 MHz, CD3OD) δ 8.74 (m, 1H), 8.00 (m, 1H), 7.85 (d, J = 7.7 Hz, 1H), 7.60 - 7.43 (m, 2H), 7.14 (t, J = 9.5 Hz, 2H), 5.63 (s, 1H), 5.06 (m, 1H), 4.64 (s, 1H), 4.52 - 4.31 (m, 3H), 4.27 - 4.05 (m, 3H), 3.97 (m, 1H), 3.92 - 3.66 (m, 6H), 3.59 (m, 2H), 3.46 (m, 2H), 3.25 (d, J = 5.3 Hz, 3H), 3.07 - 2.89 (m, 2H), 2.86 - 2.59 (m, 3H), 2.38 - 2.32 (m, 3H), 2.28 (s, 3H), 2.13 (m, 2H), 2.03 - 1.51 (m, 6H), 1.50 - 1.41 (m, 6H), 1.38 (d, J = 7.5 Hz, 3H), 0.98 (t, J = 8.7 Hz, 6H), 0.89 (t, J =6.4 Hz, 3H), 0.54 (d, J = 8.4 Hz, 3H).EXAMPLE A722. SUMMARY OF 1-ACRYLOYL-N-((2S)-1-(((63S,4S)-11-ETHYL-12-(2-((S)-1-METHOXYETHYL)PYRIDI N-3-IL)-10,10-DIMETHYL-5,7-DIOXO-61,62,63,64,65,66-HEXA-HYDRO-11H-8-OXA- 2(4,2)-MORPHOLINE-1(5,3)-INDOLE-6(1,3)-PYRDAZINECYCLOUNDECAPHANE-4-YL)AMINO)-3-METHYL-1-OXOBUTAN-2-YL)-4-FLUORO-N-METHYLPIPERIDINE-4-CARBOXAMIDE

[00483] Step 1. To a mixture of N-(4-fluoropiperidine-4-carbonyl)-N-methyl-L- (190mg, 0.73 mmol) and NaHCO3 (306 mg, 3.6 mmol) in DCM (2 mL) and H2O (1 mL) at -10 °C was added prop-2-enoyl chloride (132 mg, 1.45 mmol). The mixture was stirred at 0-5 °C for 1 h, then diluted with DCM (20 mL) and washed with H2O (20 mL × 2), brine (20 mL), and the organic layer dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give N-(1-acryloyl-4-fluoropiperidine-4-carbonyl)-N-methyl-L-valine (120 mg, 52% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C15H23FN2O4 314.2; found 315.2.

[00484] Step 2. To a mixture of (63S,4S)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexa- hydro-11H-8-oxa-2(4,2)-morpholine-1(5,3)-indole-6(1,3)-pyridazinecycloundephane-5,7-dione (153 mg, 0.24 mmol), N-(1-acryloyl-4-fluoropiperidine-4-carbonyl)-N-methyl-L-valine (106 mg, 0.34 mmol) in DMF (2 mL) at 5°C was added HATU (110 mg, 0.29 mmol) and DIPEA (468 mg, 3.6 mmol) dropwise. The mixture was stirred at 5 °C for 1 h, then purified by preparative HPLC to obtain 1-acryloyl-N-((2S)-1-(((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholine-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)-4-fluoro-N-methylpiperidine-4-carboxamide (69.5 mg, 29% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C50H69FN8O8 928.5; found 929.4; - 7.02 (m, 2H), 6.80 (dd, J = 16.8, 10.7 Hz, 1H), 6.23 (d, J = 16.8Hz, 1H), 5.77 (d, J = 10.6 Hz, 1H), 5.67 (d, J = 6.5 Hz, 1H), 4.61 (d, J = 11.1Hz, 1H), 4.44 (t, J = 15.1 Hz, 2H), 4.23 (q, J = 6.1 Hz, 1H), 4.18 - 4.10 (m, 1H), 4.09 - 4.01 (m, 1H), 3.99 - 3.83 (m, 3H), 3.83 - 3.65 (m, 4H), 3.58 - 3.46(m, 2H), 3.27 (s, 1H), 3.21 - 3.11 (m, 6H), 3.00 - 2.91 (m, 2H), 2.85 - 2.75(m, 2H), 2.73 - 2.64 (m, 1H), 2.62 - 2.54 (m, 1H), 2.36 - 2.21 (m, 2H), 2.19 -2.01 (m, 5H), 1.92 (d, J=12.7Hz, 2H), 1.79 - 1.57 (m, 2H), 1.44 (d, J=6.2Hz, 3H), 1.04 (t, J=6.3Hz, 3H), 0.98 - 0.81 (m, 6H), 0.76 (s,3H), 0.68 (s,3H). EXAMPLE377. SYNTHESIS OF (2R)-2-(((1-(4-(DIMETHYLAMINO)-4-METHYLPENT-2- INOYL)AZETIDIN-3-YL)OXY)METHYL)-N-((22S,63S,4S)-11 -ETHYL-12-(2-((S)-1 - METHOXYETHYL)PYRIDIN-3-IL)-10,10-DIMETHYL-5,7-DIOXO-61,62,63,64,65,66-HEXA- HYDRO-11H-8-OXA-2(4,2)-MORPHOLINE-1(5,3)-INDOLE-6(1,3)- PYIDAZINECYCLOUNDECAPHANE-4-YL)-3-METHYLBUTANAMIDE

[00485] Step 1. (2R)-2-(((1-(4-(dimethylamino)-4-methylpent-2-inoyl)azetidin-3-yl)oxy)methyl)-N-((22S,63S,4S)-11-ethyl-12-(2-((S)-1- methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholine-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-4-yl)-3-methylbutanamide was synthesized in a similar to (2R)-2-(((1-(4- (dimethylamino)-4-methylpent-2-inoyl)azetidin-3-yl)oxy)methyl)-N-((22S,63S,4S)- 12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2- trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholine-1(5,3)-indole-6(1,3)-pyridazinecycloundephane-4-yl)-3-methylbutanamide except(22S,63S,4S)-4-amino-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-11- (2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholine-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-5,7-dione was replaced with (22S,63S,4S)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholine-1(5,3)- indole-6(1,3)-pyridazinecycloundecaphane-5,7-dione and acid 3-({1-[4-(dimethylamino)-4-methylpent-2-inoyl]azetidin-3-yl}oxy)propanoic acid was replaced by (2R)-2-[({1-[4-(dimethylamino)-4-methylpent-2-inoyl]azetidin-3-yl}oxy)methyl]-3-methylbutanoic acid to give the desired product (25.6 mg, 26% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C52H74N8O8 938.6; found 939.5; 1H NMR (400 MHz, CD3OD) δ 8.72 (dd, J = 4.8, 1.5 Hz, 1H), 7.97 (dd, J = 20.0, 6.8 Hz, 1H), 7.87 (dd, J = 5.8, 2.6 Hz, 1H), 7.54 - 7.51 (m, 1H), 7.41 (d, J = 8.9 Hz, 1H), 7.14 (dd, J = 36.0, 10.4Hz, 2H), 5.65 (s, 1H), 4.49 - 4.33 (m, 3H), 4.27 - 4.08 (m, 4H), 3.96 (d, J = 8.6 Hz, 2H), 3.87 (dd, J = 10.8, 3.6 Hz, 2H), 3.79 (dd, J = 10.8, 7.7 Hz, 3H), 3.69 - 3.58 (m, 3H), 3.43 (dd, J = 23.9, 11.7 Hz, 2H), 3.17 (d, J = 21.9 Hz, 3H), 3.00 - 2.95 (m, 1H), 2.70 (t, J = 14.0 Hz, 8H), 2.34 - 2.24 (m, 1H), 2.05 (d, J = 34.4 Hz, 3H), 1.92 - 1.82 (m, 2H), 1.69 - 1.62 (m, 5H), 1.57 (d, J = 11.5 Hz, 3H), 1.45 (d, J = 6.2Hz, 3H), 1.33 (d, J = 12.6Hz, 1H), 1.05 - 0.93 (m, 10H), 0.80 (d, J = 9.8Hz, 3H), 0.64 (d, J = 12.2Hz, 2H).EXAMPLE A643. SYNTHESIS OF (2R)-2-(((1-(4-(DIMETHYLAMINO)-4-METHYLPENT-2- INOYL)PIPERIDIN-4-YL)OXY)METHYL)-N-((22S,63S,4S)-11 -ETHYL-12-(2-((S)-1 - METHOXYETHYL)PYRIDIN-3-IL)-10,10-DIMETHYL-5,7-DIOXO-61,62,63,64,65,66-HEXA-HYDRO- 11H-8-OXA-2(4,2)-MORPHOLINE-1(5,3)-INDOLE-6(1,3)-PYRIDAZINECYCLOUNDECAFAN- 4-IL)-3-METHYLBUTANAMIDE

[00486] Step 1. A mixture of 1-(1-methylphenyl)piperidin-4-yl methanesulfonate (2 g, 7.4 mmol) and tert-butyl (2R)-2-(hydroxymethyl)-3-methylbutanoate (1.39 g, 7.4 mmol) was stirred at 120 °C for 1 h, then purified by silica gel column chromatography to give tert-butyl (R)-2-(((1-benzylpiperidin-4-yl)oxy)methyl)-3-methylbutanoate (800 mg, 28% yield) as an oil. LCMS (ESI): m/z [M+H] calcd for C22H35NO3 361.3; found 362.3.

[00487] Step 2. A mixture of tert-butyl (R)-2-(((1-benzylpiperidin-4-yl)oxy)methyl)-3-methylbutanoate (700 mg, 1.9 mmol), 10% wet Pd/C (411 mg, 3.9 mmol), and 20% wet Pd(OH)2/C (542 mg, 3.9 mmol) in THF (30 mL) was stirred under an H2 atmosphere (15 psi) for 16 h. The mixture was filtered and the filtrate was concentrated under reduced pressure to give tert-butyl (R)-3-methyl-2-((piperidin-4-yloxy)methyl)butanoate (440 mg, 80% yield) as an oil. LCMS (ESI): m/z [M+H] calculated for C15H29NO3 271.2; found 272.2.

[00488] Step 3. To a mixture of tert-butyl (R)-3-methyl-2-((piperidin-4-yloxy)methyl)butanoate (440 mg, 1.6 mmol), 4-(dimethylamino)-4-methylpent-2-ynoic acid (3.77 g, 24.3 mmol), and DIPEA (2.09 g, 16.2 mmol) in DMF (50 mL) at 0 °C was added T3P (2.57 g, 8.1 mmol). The mixture was stirred at 0 °C for 1 h, then poured into H2O (50 mL) and extracted with EtOAc (50 mL × 3). The combined organic layers were washed with brine, concentrated under reduced pressure, and the residue purified by silica gel column chromatography to give tert-butyl (R)-2-(((1-(4-(dimethylamino)-4-methylpent-2-ynoyl)piperidin-4-yl)oxy)methyl)-3-methylbutanoate (190 mg, 27% yield) as an oil. LCMS (ESI): m/z [M+H] calcd for C23H40N2O4 408.3; found 409.4.

[00489] Step 4. To a mixture of tert-butyl (R)-2-(((1-(4-(dimethylamino)-4-methylpent-2-ynoyl)piperidin-4-yl)oxy)methyl)-3-methylbutanoate (180 mg, 0.47 mmol) in DCM (2 mL) was added TFA (1 mL). The mixture was stirred at room temperature for 1 h, then concentrated under reduced pressure to give (R)-2-(((1-(4-(dimethylamino)-4-methylpent-2-ynoyl)piperidin-4-yl)oxy)methyl)-3-methylbutanoic acid (170 mg, 98% yield) as an oil. LCMS (ESI): m/z [M+H] calcd for C19H32N2O4 352.2; found 353.2.

[00490] Step 5. (2R)-2-(((1-(4-(dimethylamino)-4-methylpent-2-inoyl)piperidin-4-yl)oxy)methyl)-N-((22S,63S,4S)-11-ethyl-12-(2-((S)-1- methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholine-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-4-yl)-3-methylbutanamide was synthesized in a similar way to (2R)-2- (((1-(4-(dimethylamino)-4-methylpent-2-inoyl)azetidin-3-yl)oxy)methyl)-N- ((22S,63S,4S)-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11- (2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholine- 1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-4-yl)-3-methylbutanamide except (22S,63S,4S)-4-amino-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)- Morpholine-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-5,7-dione was replaced by (22S,63S,4S)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3- (2R)-2-[({1-[4-(dimethylamino)-4-methylpent-2-ynoyl]piperidin-4-yl}oxy)methyl]-3-methylbutanoic acid (101 mg, 42% yield) as a solid. LCMS (ESI): m/z [M+H] calcd. for C54H78N8O8 966.6; found 969.5;1H NMR (400 MHz, CD3OD) δ 8.70 (d, J = 4.8 Hz, 1H), 7.81 - 7.76 (m, 1H), 7.56 - 7.46 (m, 1H), 7.43 - 7.34 (m, 1H), 7.24 - 7.01 (m, 2H) 3.47 - 3.34 ( m, 2H), 3.26 - 3.15 (m, 3H), 2.98 - 2.57 (m, 5H), 2.37 - 2.30 (m, 3H), 2.27 - 2.18 (m, 4H), 2.15 - 2.02 (m, 2H), 2.00 - 1.80 (m, 4H), 1.78 - 1.71 (m, 2H), 1.68 - 1.55 (m, 3H), 1.49 - 1.37 (m, 6H), 1.35 - 1.28 (m, 3H), 1.05 - 0.92 (m, 9H), 0.85 - 0.72 (m, 3H), 0.68 - 0.51 (m, 3H).EXAMPLE A328. SYNTHESIS OF TWO ATROPISOMERS OF (3S)-1 -ACRYLOYL-N-((2S)-1 - (((63S,4S)-11 -ETHYL-12-(2-((S)-1 -METHOXYETHYL)-5-(4-METHYLPIPERAZIN-1 -IL)PYRIDIN-3-IL)-10,10-DIMETHYL-5,7-DIOXO-61,62,63,64,65,66-HEXA-HYDRO-11H-8-OXA-1(5,3)- INDOLA-6(1,3)-PYRIDAZINE-2(1,3)-BENZENACYCLOUNDECAPHANE-4-YL)AMINO)-3-METHYL-1-OXOBUTAN-2-YL)-N-METHYLPYRROLIDINE-3-CARBOXAMIDE

[00491] Step 1. To a mixture of 3-bromo-5-iodo-2-[(1 S)-1-methoxyethyl]pyridine (2.20 g, 6.4 mmol) and tert-butyl piperazine-1-carboxylate (1.20 g, 6.4 mmol) in toluene (50 mL) under an Ar atmosphere were added tBuONa (0.74 g, 7.7 mmol) and portionwise addition of Pd2(dba)3 (0.59 g, 0.64 mmol), followed by portionwise addition of Xantphos (0.74 g, 1.3 mmol). The mixture was heated to 100 °C and stirred for 16 h, then H2O was added and the mixture extracted with EtOAc (400 mL × 3). The combined organic layers were washed with brine (150 mL × 3), dried over anhydrous Na 2 SO 4 , and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by preparative HPLC to give tert-butyl 4-[5-bromo-6-[(1 S)-1-methoxyethyl]pyridin-3-yl]piperazine-1-carboxylate (1.7 g, 61% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C 17H 26BrN 3 O 3 399.1; found 400.1.

[00492] Step 2. To a mixture of tert-butyl 4-[5-bromo-6-[(1 S)-1-methoxyethyl]pyridin-3-yl]piperazine-1-carboxylate (1.76 g, 4.4 mmol) and 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (1.67 g, 6.6 mmol) in toluene (18 mL) under an Ar atmosphere were added KOAc (0.95 g, 9.7 mmol) and Pd(PPh3)2Cl2 (0.31 g, 0.44 mmol) in portions. The mixture was heated to 80 °C and stirred for 16 h, then diluted with H2O and the mixture extracted with EtOAc (500 mL × 3). The combined organic layers were washed with brine (100 mL × 3), dried over anhydrous Na 2 SO 4 , and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give tert-butyl 4-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl]piperazine-1-carboxylate (1.4 g, 68% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C 23H 38 BN 3 O 5 447.4; found 448.2.

[00493] Step 3. To a mixture of tert-butyl((63S,4S)-12-iodo-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (1.0 g, 1.5 mmol) in DCM (10 mL) at 0 °C under a N2 atmosphere was added TFA (5.0 mL, 67.3 mmol) in portions. The mixture was stirred at 0 °C for 1 h, then concentrated under reduced pressure and azeotropically dried with toluene (3 mL × 3) to give (63S,4S)-4-amino-12-iodo-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-5,7-dione (1.0 g), which was used directly in the next step without further purification. LCMS (ESI): m/z [M+H] calcd for C27H31IN4O3 586.1; found 587.3.

[00494] Step 4. To a mixture of (63S,4S)-4-amino-12-iodo-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-5,7-dione (1.0 g, 1.7 mmol) in DMF (15 mL) at 0 C under a N 2 atmosphere were added DIPEA (2.20 g, 17.0 mmol) and (2S)-2-[[(benzyloxy)carbonyl](methyl)amino]-3-methylbutanoic acid (0.90 g, 3.4 mmol) in portions, followed by COMU (1.10 g, 2.6 mmol) in portions over 10 min. The mixture was stirred at 0 °C for 1.5 h, then diluted with H2O and the mixture was extracted with EtOAc (300 mL × 3). The combined organic layers were washed with brine (100 mL × 3), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative HPLC to give benzyl ((2S)-1-(((63S,4S)-12-iodo-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl) (methyl)carbamate (790 mg, 53% yield) as a solid.

[00495] Step 5. To a mixture of benzyl ((2S)-1-(((63S,4S)-12-iodo-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H- 8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundephane-4-yl)amino)-3-methyl-1-oxobutan-2-yl) (methyl)carbamate (480 mg, 0.58 mmol) and tert-butyl 4-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2- English: To 1,4-yl)pyridin-3-yl]piperazine-1-carboxylate (309 mg, 0.69 mmol) in 1,4-dioxane (8.0 mL) and H2O (1.6 mL) under an Ar atmosphere were added K2CO3 (199 mg, 1.4 mmol) and Pd(dppf)Cl2 (42 mg, 0.06 mmol) in portions. The mixture was heated to 70 °C and stirred for 16 h, then diluted with H2O and extracted with EtOAc (200 mL × 3). The combined organic layers were washed with brine (150 mL × 3), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain tert-butyl 4-(5-((63S,4S)-4-((S)-2-(((benzyloxy)carbonyl)(methyl)amino)-3-methylbutanamido)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-12-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate (335 mg, 51% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C58H74N8O9 1026.6; found 1027.4.

[00496] Step 6. To a mixture of tert-butyl 4-(5-((63S,4S)-4-((S)-2-(((benzyloxy)carbonyl)(methyl)amino)-3-methylbutanamido)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-12-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate (335 mg, 0.33 mmol) in DMF (5 mL) at 0 °C under a N2 atmosphere was added Cs2CO3 (234 mg, 0.72 mmol) and iodoethane (102 mg, 0.65 mmol) in portions. The mixture was warmed to room temperature and stirred for 16 h, then diluted with H2O and the mixture extracted with EtOAc (100 mL × 3). The combined organic layers were washed with brine (50 mL × 3), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative TLC to give tert-butyl 4-(5-((63S,4S)-4-((S)-2-(((benzyloxy)carbonyl)(methyl)amino)-3-methylbutanamido)-11-ethyl-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-12-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate (320 mg, 84% yield) as a light yellow solid. LCMS (ESI): m/z [M+H] calculated for C60H78N8O9 1054.6; found 1055.8.

[00497] Step 7. A mixture of tert-butyl 4-(5-((63S,4S)-4-((S)-2-(((benzyloxy)carbonyl)(methyl)amino)-3-methylbutanamido)-11-ethyl-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-12-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate (320 mg) in xx M HCl in 1,4-dioxane (3.0 mL) at 0 C under an atmosphere of N 2 was stirred at room temperature for 2 h, then concentrated under reduced pressure to give benzyl ((2S)-1-(((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)-5-(piperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazin-2(1,3)-benzenacycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamate, which was directly used in the next further purification step. LCMS (ESI): m/z [M+H] calcd for C55H70N8O7 954.5; found 955.3.

[00498] Step 8. To a mixture of benzyl ((2S)-1-(((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)-5-(piperazin-1-yl)pyridin-3-yl)-10,10- dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl) (methyl)carbamate (320 mg, 0.34 mmol) and HCHO (60 mg, 2.0 mmol) in MeOH (3.0 mL) at 0 C under a N2 atmosphere NaCNBH3 (42 mg, 0.67 mmol) and AcOH (60 mg, 1.0 mmol) were added portionwise. The mixture was warmed to room temperature and stirred for 2 h, then diluted with H2O and the mixture extracted with DCM/MeOH (5:1) (200 mL × 3). The combined organic layers were washed with brine (100 mL × 3), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative HPLC to give benzyl ((2S)-1-(((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl) (methyl)carbamate (160 mg, 59% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C56H72N8O7 968.6; found 969.6.

[00499] Step 9. To a mixture of benzyl ((2S)-1-(((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)- 10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundephane-4-yl)amino)-3-methyl-1-oxobutan-2-yl) (methyl)carbamate (160 mg, 0.17 mmol) in toluene (10 mL) and MeOH (1.0 mL) Pd/C (130 mg, 1.2 mmol) in portions. The mixture was evacuated and recharged with H2 (x 3), then stirred under an atmosphere of H2 for 16 h. The mixture was filtered and the filtrate was concentrated under reduced pressure to give (2S)-N-((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)-3-methyl-2-(methylamino)butanamide (140 mg), which was used directly in the next step without further purification. LCMS (ESI): m/z [M+H] calcd for C48H66N8O5 834.5; found 835.5.

[00500] Step 10. To a mixture of (2S)-N-((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazin-2(1,3)-benzenacycloundecaphane-4-yl)-3-methyl-2-(methylamino)butanamide (140 mg, 0.17 mmol) in ACN (2.0 mL) at 0 °C under a N 2 atmosphere were added DIPEA (433 mg, 3.35 mmol), ethyl acetate (4.3 mg, 1.2 mmol), and diethyl ether (4.3 mg, 1.2 mmol). (3S)-1-(prop-2-enoyl)pyrrolidine-3-carboxylic acid (57 mg, 0.34 mmol) portionwise and CIP (70 mg, 0.25 mmol) portionwise over 10 min. The mixture was stirred at 0 C for 1.5 h, then H2O was added and the mixture extracted with EtOAc (150 mL × 3). The combined organic layers were washed with brine (100 mL × 3), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by preparative HPLC to obtain two atropisomers of (3S)-1-acryloyl-N-((2S)-1-(((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)-N-methylpyrrolidine-3-carboxamide (40 mg, 24% yield) as a solid and (20 mg, 12% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C56H75N9O7 985.6; found 986.7; 1H NMR (400 MHz, DMSO-d6) 8.47 (t, J = 2.1 Hz, 1H), 8.00 (d, J = 4.7 Hz, 1H), 7.78 - 7.59 (m, 3H), 7.58 - 7.48 (m, 1H), 7.42 - 7.30 (m, 1H), 7.23 (dq, J = 8.0, 4.0, 3.5 Hz, 1H), 7.15 - 7.03 (m, 1H), 6.75 - 6.50 (m, 1H), 6.18 (dt, J = 16.8, 2.7 Hz, 1H), 5.70 (tt, J = 9.3, 2.7 Hz, 1H), 5.48 - 5.23 (m, 1H), 5.06 (dd, J = 31.1, 12.3 Hz, 1H), 4.74 (dd, J = 11.0, 4.3 Hz, 1H), 4.33 - 4.15 (m, 2H), 4.01 (ddd, J = 36.1, 12.6, 7.6 Hz, 2H), 3.91 - 3.56 (m, 6H), 3.52 - 3.39 (m, 2H), 3.31 - 3.28 (m, 2H), 3.24 (d, J = 5.7 Hz, 4H), 3.06 (s, 4H), 2.93 (d, J = 9.8 Hz, 2H), 2.81 (d, J = 5.4 Hz, 3H), 2.47 - 2.43 (m, 4H), 2.22 (s, 4H), 2.09 (tq, J = 12.0, 7.4, 6.6 Hz, 3H), 1.81 (s, 1H), 1.74 (d, J = 11.7 Hz, 1H), 1.56 (d, J = 11.7 Hz, 1H), 1.20 (dd, J = 6.3, 1.5 Hz, 3H), 1.10 (td, J = 7.2, 2.4 Hz, 3H), 1.00 - 0.86 (m, 6H), 0.86 - 0.72 (m, 3H), 0.54 (d, J = 3.5 Hz, 3H) and LCMS (ESI): m/z [MH] calculated for C56H75N9O7 985.6; found 984.4; 1H NMR (400 MHz, DMSO-d6) δ 8.46 (d, J = 3.0 Hz, 2H), 7.98 (s, 1H), 7.89 - 7.83 (m, 1H), 7.76 - 7.57 (m, 3H), 7.24 (s, 2H), 7.07 (s, 1H), 6.70 - 6.58 (m, 1H), 6.17 (d, J = 16.5 Hz, 1H), 5.73 - 5.67 (m, 1H), 5.36 - 5.30 (m, 1H), 4.31 - 3.97 (m, 6H), 3.83 - 3.77 (m, 2H), 3.74 - 3.49 (m, 6H), 3.48 - 3.41 (m, 1H), 3.40 - 3.37 (m, 2H) 2.44 (m, 4H), 2.22 (s, 3H), 2.04 (d, J = 26.1 Hz, 3H), 1.85 - 1.79 (m, 1H), 1.67 - 1.55 (m, 2H), 1.35 (d, J = 6.1 Hz, 3H), 1.27 -1.22 (m, 1H), 1.05 - 0.93 (m, 4H), 0.89 (d, J = 6.9 Hz, 2H), 0.79 (d, J = 12.4Hz, 5H), 0.73 (d, J = 6.5 Hz, 1H), 0.56 (s, 3H).EXAMPLE A542. SYNTHESIS OF 1-(4-(DIMETHYLAMINO)-4-METHYLPENT-2-INOYL)-4-FLUORO- N-((2S)-1 -(((63S,4S)-12-(2-((S)-1 -METHOXYETHYL)-5-(4-METHYLPIPERAZIN-1 -YL)PYRIDIN- - BENZENACYCLOUNDECAPHANE-4-YL)AMINO)-3-METHYL-1-OXOBUTAN-2-YL)-N-METHYLPYPERIDINE-4-CARBOXAMIDE

[00501] Step 1. To a solution of (S)-3-bromo-5-iodo-2-(1-methoxyethyl)pyridine (15 g, 43.86 mmol) and benzyl piperazine-1-carboxylate (8.7 g, 39.48 mmol) in toluene (150 mL) at 0 °C was added cesium carbonate (71.46 g, 219.32 mmol), BINAP (0.55 g, 0.88 mmol), and palladium acetate (0.49 g, 2.19 mmol) in portions. The reaction mixture was stirred at 90 °C for 12 h under an argon atmosphere. The resulting mixture was cooled to room temperature, filtered, and the filter cake was washed with EtOAc (150 mL × 3). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give benzyl (S)-4-(5-bromo-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate (16 g, 84% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C44H58N6O7 433.1; found 434.0.

[00502] Step 2. To a stirred solution of benzyl (S)-4-(5-bromo-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate (22.7 g, 52.26 mmol), bis(pinacolato)diboron (19.91 g, 78.4 mmol) in toluene (230 mL) at 0 °C was added potassium acetate (12.82 g, 130.66 mmol) and Pd(dppf)C^DCM (4.26 g, 5.23 mmol) in portions. The reaction mixture was stirred at 90 °C for 6 h under an argon atmosphere. The resulting mixture was filtered, and the filter cake was washed with EtOAc (200 mL x 3). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to afford benzyl (S)-4-(6-(1-methoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)piperazine-1-carboxylate (14.7 g, 58% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C26H36BN3O5 481.3; found 482.3.

[00503] Step 3. To a stirred solution of 5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-iodo-1H-indole (17.46 g, 27 mmol) in 1,4-dioxane (150 mL) and H2O (30 mL) at 0 °C were added potassium carbonate (9.33 g, 67.51 mmol) and Pd(dppf)Cl2 DCM (2.2 g, 2.7 mmol) in portions, followed by benzyl (S)-4-(6-(1-methoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)piperazine-1-carboxylate (13 g, 27 mmol). The reaction mixture was stirred at 70 °C for 12 h under an argon atmosphere. The resulting mixture was cooled to room temperature and quenched with H2O, then extracted with EtOAc (200 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to afford benzyl (S)-4-(5-(5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate (20 g, 84.7% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C49H57BN4O4Si 873.2; found 873.3.

[00504] Step 4. To a mixture of benzyl (S)-4-(5-(5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate (19 g, 21.74 mmol) and Cs2CO3 (49.58 g, 152.17 mmol) in DMF (190 mL) at 0 °C under argon atmosphere, 2,2,2-trifluoroethyl trifluoromethanesulfonate (50.46 g, 217.39 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 12 h under an argon atmosphere, then quenched with H2O, extracted with EtOAc (200 mL × 3). The combined organic layers were washed with brine (200 mL × 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to afford benzyl (S)-4-(5-(5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate (17.6 g, 84.7% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C51H58BF3N4O4Si 954.2; found 955.3.

[00505] Step 5. To a solution of benzyl (S)-4-(5-(5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate (18 g, 18.83 mmol), was added TBAF in THF (180.0 mL) at 0 °C. The reaction mixture was stirred at 40 °C for 12 h under an argon atmosphere, then quenched with cold H2O. The resulting mixture was extracted with EtOAc (200 mL × 3). The combined organic layers were washed with brine (200 mL × 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to afford benzyl (S)-4-(5-(5-bromo-3-(3-hydroxy-2,2-dimethylpropyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate (7.8 g, 57.7% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C35H40BrF3N4O4 716.2; found 717.1.

[00506] Step 6. A solution of benzyl (S)-4-(5-(5-bromo-3-(3-hydroxy-2,2-dimethylpropyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate (1 g, 1.39 mmol) in 1,4-dioxane (10 mL) and H2O (2 mL) was added methyl (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanoyl)hexahydropyridazine-3-carboxylate (1.08 g, 2.09 mmol), potassium carbonate English: (481.47 mg, 3.48 mmol) and Pd(dtbpf)Cl2 (181.64 mg, 0.28 mmol) in portions at 0 °C. The reaction mixture was stirred at 70 °C for 3 h under an argon atmosphere. The resulting mixture was cooled to room temperature, then quenched with H2O and extracted with EtOAc (50 mL × 3). The combined organic layers were washed with brine (20 mL × 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography to give methyl (S)-1-((S)-3-(3-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-(3-hydroxy-2,2-dimethylpropyl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate (1.1 g, 77% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C55H68F3N7O9 1027.5; found 1028.3.

[00507] Step 7. To a solution of methyl (S)-1-((S)-3-(3-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-(3-hydroxy-2,2-dimethylpropyl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-ylphenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate (1.1 g, 1.07 mmol) in THF (8 mL) and H2O (2 mL) at 0 °C, LiOH (2.2 mL, 1 M aqueous) was added dropwise under an argon atmosphere. The reaction mixture was stirred for 2 h and then concentrated under reduced pressure. The residue was acidified to pH 5 with citric acid (1 M) and extracted with EtOAc (20 mL × 3). The combined organic layers were concentrated under reduced pressure. The residue was purified by reversed-phase chromatography to give (S)-1-((S)-3-(3-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-(3-hydroxy-2,2-dimethylpropyl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylic acid (750 mg, 69% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C54H66F3N7O9 1013.5; found 1014.3.

[00508] Step 8. To a solution of (S)-1-((S)-3-(3-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-(3-hydroxy-2,2-dimethylpropyl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylic acid (0.75 g, 0.74 mmol) in DCM (75 mL) at 0 °C were added portionwise HOBT (0.5 g, 3.7 mmol), DIPEA (3.82 g, 29.58 mmol), and EDCI (4.25 g, 22.19 mmol). mmol) at 0 °C. The reaction mixture was stirred at room temperature for 12 h under an argon atmosphere. The resulting mixture was concentrated under reduced pressure and extracted with EtOAc (100 mL × 3). The combined organic layers were washed with brine (50 mL × 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography to afford benzyl 4-(5-((63S,4S)-4-((tert-butoxycarbonyl)amino)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-12-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate (0.5 g, 67.9% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C54H64F3N7O8 995.5; found 996.3.

[00509] Step 9. To a mixture of benzyl 4-(5-((63S,4S)-4-((tert-butoxycarbonyl)amino)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-12-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate (500 mg, 0.5 mmol) in MeOH (15 mL) at 0 °C was added paraformaldehyde (135.64 mg, 1.5 mmol) and Pd/C (750 mg) in portions. The reaction mixture was stirred at room temperature for 12 h under a hydrogen atmosphere. The resulting mixture was filtered, and the filter cake was washed with EtOAc (50 mL × 5). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography to afford tert-butyl ((63S,4S)-12-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (350 mg, 79.6% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C47H60F3N7O8 875.5; found 876.5.

[00510] Step 10. To a solution of tert-butyl((63S,4S)-12-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-4-yl)carbamate (300 mg, 0.34 mmol) in DCM (2 mL) at 0 °C was added dropwise HCl in 1,4-dioxane (1 mL, 4M, 4 mmol). The reaction mixture was stirred at room temperature for 2 h, then concentrated under reduced pressure to give (63S,4S)-4-amino-12-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(1,3)-benzenacycloundecaphane-5,7-dione hydrochloride (350 mg, crude) as a solid. LCMS (ESI): m/z [M+H] calculated for C42H52F3N7O4 775.4; found 766.4.

[00511] Step 11. To a hydrochloride solution of (63S,4S)-4-amino-12-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10- dimethyl-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(1,3)-benzenecycloundecaphane-5,7-dione (150 mg, 0.19 mmol) and N-(1-(4-(dimethylamino)-4-methylpent-2-inoyl)-4-fluoropiperidine-4-carbonyl)-N-methyl-L-valine (154 English:mg, 0.39 mmol) in DMF (2 mL) at 0 °C, a mixture of DIPEA (1 g, 7.72 mmol) and HATU (110 mg, 0.29 mmol) in DMF (0.2 mL) was added dropwise. The reaction mixture was stirred at 0 °C for 2 h under an argon atmosphere, then quenched with H2O. The resulting mixture was extracted with EtOAc (20 mL × 3). The combined organic phase was washed with brine (10 mL × 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by reverse phase chromatography to give 1-(4-(dimethylamino)-4-methylpent-2-inoyl)-4-fluoro-N-((2S)-1-(((63S,4S)-12-(2-((S)-1-methoxyethyl)-5-(4- methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)- 61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine- 2(1,3)-benzenecycloundephane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)-N- methylpiperidine-4-carboxamide (39.5 mg, 17% yield) as a solid. 1H NMR (400 MHz, DMSO-d6) δ 8.48 (d, J = 2.9 Hz, 1H), 8.32 (t, J = 7.3 Hz, 1H), 7.97 (s, 1H), 7.82-7.69 (m, 3H), 7.66 (t, J = 7.8 Hz, 1H), 7.33-7.07 (m, 3H), 5.50 (dd, J = 16.7, 8.6 Hz, 1H), 5.33 (t, J = 9.2 Hz, 1H), 5.16 (d, J = 12.2 Hz, 1H), 4.94-4.80 (m, 1H), 4.64 (d, J= 10.8Hz, 1H), 4.33-4.16(m, 3H), 4.12-4.02 (m, 2H), 3.71-3.50 (m, 3H), 3.25 (s, 3H), 3.21-3.16 (m, 3H), 3.14 -3.05 (m, 1H), 2.96 (t, J = 4.7 Hz, 4H), 2.84 (s, 1H), 2.82-2.72 (m, 2H), 2.59-2.53 (m, 1H), 2.47-2.40 (m, 4H), 2.22 (d, J = 2.9 Hz, 9H), 2.18-2.12 (m, 2H), 2.111.99 (m, 3H), 1.87-1.78 (m, 1H) 0.77 (d, J = 6.6 Hz, 2H), 0.38 (s, 3H). LCMS (ESI): m/z [M+H] calculated for C44H58N6O7 1154.6; found 1155.7.EXAMPLE A735. SUMMARY OF 2-ACRYLOYL-N-((2S)-1-(((63S,4S,Z)-11-ETHYL-12-(2-((S)-1-METHOXYETHYL)PYRIDIN-3-IL)-10,10-DIM ETHYL-5,7-DIOXO-61,62,63,64,65,66-HEXA-HYDRO-11H-8-OXA-2(4,2)-THIAZOLA-1(5,3)-INDOLE-6(1,3)- PYIDAZINECYCLOUNDECAPHANE-4-YL)AMINO)-3-METHYL-1-OXOBUTAN-2-YL)-N-METHYL-5-OXA-2,9-DIAZAESPIRO[3,5]NONANE-9-CARBOXAMIDE

[00512] Step 1. To a stirred mixture of BTC (425.2 mg, 1.448 mmol) in DCM (10 mL) was added dropwise pyridine (1.04 g, 13.16 mmol) and tert-butyl 5-oxa-2,9-diazaspiro[3.5]nonane-2-carboxylate (1 g, 4.39 mmol), the reaction mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure to give crude tert-butyl 9-(chlorocarbonyl)-5-oxa-2,9-diazaspiro[3.5]nonane-2-carboxylate.

[00513] Step 2. To a stirred solution of tert-butyl 9-(chlorocarbonyl)-5-oxa-2,9-diazaspiro[3.5]nonane-2-carboxylate (2.5 g, crude) in MeCN (20 mL) were added dropwise pyridine (1.04 g, 13.16 mmol) and benzyl (2S)-3-methyl-2-(methylamino)butanoate (970.66 mg, 4.38 mmol) at room temperature. The reaction mixture was stirred at 80 °C for 12 h and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to afford tert-butyl (S)-9-((1-(benzyloxy)-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)-5-oxa-2,9-diazaspiro[3.5]nonane-2-carboxylate (783 mg, 37.6% yield, two steps) as an oil. LCMS (ESI): m/z [M+H] calcd for C25H37N3O6 475.3; found 476.3.

[00514] Step 3. A solution of tert-butyl (S)-9-((1-(benzyloxy)-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)-5-oxa-2,9-diazaspiro[3.5]nonane-2-carboxylate (783 mg, 1.65 mmol) and 10 wt % palladium on carbon (226.29 mg) in THF (10 mL) was stirred for 2 h at 50 °C under a hydrogen atmosphere. The resulting mixture was cooled to room temperature, filtered, and the filter cake was washed with MeCN (10 mL × 3). The filtrate was concentrated under reduced pressure to give N-(2-(tert-butoxycarbonyl)-5-oxa-2,9-diazaspiro[3.5]nonane-9-carbonyl)-N-methyl-L-valine (591 mg, 98.8% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C18H31N3O6 385.2; found 386.3.

[00515] Step 4. To a stirred solution of intermediate 2 (731 mg, 1.16 mmol) and DIPEA (2.25 g, 17.38 mmol) in MeCN (50 mL) was added CIP (644.31 mg, 2.32 mmol) and N-(2-(tert-butoxycarbonyl)-5-oxa-2,9-diazaspiro[3.5]nonane-9-carbonyl)-N-methyl-L-valine (446.68 mg, 1.16 mmol) at room temperature. The reaction mixture was stirred for 2 h and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give tert-butyl 9-(((2S)-1-(((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro- 11H-8-oxa-2(4,2)-thiazole-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)-5-oxa-2,9-diazaspiro[3,5]nonane-2-carboxylate (752 mg, 65% yield) as solid. LCMS (ESI): m/z [M+H] calculated for C52H71N9O9S 997.5; found 996.6.

[00516] Step 5. To a stirred solution of tert-butyl 9-(((2S)-1-(((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo- 61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-thiazola-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)- 5-oxa-2,9-diazaspiro[3,5]nonane-2-carboxylate (752 mg, 0.75 mmol) in DCM (40 mL) was added TFA (10 mL) in portions at room temperature. The reaction mixture was stirred for 2 h and then concentrated under reduced pressure. To the residue was added saturated aqueous sodium bicarbonate (100 mL) and DCM (100 mL). The aqueous layer was separated and extracted with DCM (100 mL × 2). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give N-((2S)-1-(((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-thiazole-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)-N-methyl-5-oxa-2,9-diazaspiro[3,5]nonane-9-carboxamide (587 mg, 87% yield) as solid. LCMS (ESI): m/z [M+H] calcd for C47H63N9O7S 897.5; found 898.4.

[00517] Step 6. A stirred solution of N-((2S)-1-(((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-thiazole-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)-N-methyl-5-oxa-2,9-diazaspiro[3,5]nonane-9-carboxamide (586 mg, 0.65 mmol) in MeCN (10 mL) was added acrylic acid (47 mg, 0.5 mmol) 0.65 mmol), DIPEA (421 mg, 3.26 mmol), CIP (362 mg, 1.3 mmol). The reaction mixture was stirred for 12 h and concentrated under reduced pressure. The residue was purified by reversed-phase chromatography to generate 2-acryloyl-N-((2S)-1-(((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8- oxa-2(4,2)-thiazole-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)-N-methyl-5-oxa-2,9-diazaspiro[3,5]nonane-9-carboxamide (194 mg, 27% yield) as a solid. 1H NMR (400 MHz, DMSO-d6) δ 8.78 (dd, J = 4.8, 1.7 Hz, 1H), 8.48 (d, J = 1.6 Hz, 2H), 7.85-7.65 (m, 3H), 7.65- 7.42(m,2H), 6.30 (mm, 1H), 6.10 (m, J = 17.0, 1H), 5.78-5.50 (m, J = 10.3, 2H), 5.10 (dd, 1H), 4.40-3.80 (m, 14H), 3.60 - 3.10 (m, 10H), 2.94 (d, J = 14.5 Hz, 1H), 2.85 (s, 4H), 2.42 (dd, 1H), 2.07 (dd, 2H), 1.80 (s, 2H), 1.55 (s, 3H), 1.32 (d, 3H), 0.95 - 0.75 (m, 12H), 0.33 (s, 3H). LCMS (ESI): m/z [M+H] calculated for C50H65N9O8S 951.5; found 952.6.EXAMPLE A720. SYNTHESIS OF (2R)-2-(((1-(4-(DIMETHYLAMINO)-4-METHYLPENT-2- INOYL)AZETIDIN-3-YL)OXY)METHYL)-N-((63S,4S,Z)-11 -ETHYL-12-(2-((S)-1 - METHOXYETHYL)PYRIDIN-3-IL)-21,10,10-TRIMETHYL-5,7-DIOXO-61,62,63,64,65,66-HEXA-HYDRO-1 1H,21H-8-OXA-1(5,3)-INDOLA-6(1,3)-PYRDAZINE-2(5,3)-TRIAZOLACYCLOUNDECAPHANE-4-YL)-3-METHYLBUTANAMIDE

[00518] Step 1. To a stirred solution of methyl 1-methyl-1,2,4-triazole-3-carboxylate (7.0 g, 49.60 mmol) in CCl4 (70 mL) was added NBS (13.24 g, 74.40 mmol) and AIBN (11.40 g, 69.44 mmol) in portions at 25 °C under an argon atmosphere. The resulting mixture was stirred for 24 h at 80 °C. The resulting mixture was filtered, the filtrate was cooled to 20 °C and kept at 20 °C for 30 min. The resulting mixture was filtered. The filter cake was washed with H2O (3 x 50 mL) and petroleum ether (3 x 100 mL). The filter cake was dried under reduced pressure. This gave methyl 5-bromo-1-methyl-1,2,4-triazole-3-carboxylate (10 g, crude) as a light yellow solid. LCMS (ESI): m/z [M+H] calculated for C5H6BrN3O2 219.0; found 219.9.

[00519] Step 2. To a stirred solution of methyl 5-bromo-1-methyl-1,2,4-triazole-3-carboxylate (10.0 g, 45.50 mmol) in MeOH (150.0 mL) and H2O (30.0 mL) was added NaBH4 (6.88 g, 181.80 mmol) in portions at -5 °C under a nitrogen atmosphere. The resulting mixture was stirred for 2 h at 0~10 °C. The desired product could be detected by LCMS. The reaction was quenched with brine (100 mL) at 0 °C. The resulting mixture was extracted with petroleum ether (100 mL). The aqueous layer was separated and filtered. The filter cake was washed with MeOH (2 x 50 mL). The filtrate was concentrated under reduced pressure to afford (5-bromo-1-methyl-1,2,4-triazol-3-yl)methanol (6 g, crude) as a light yellow solid. LCMS (ESI): m/z [M+H] calculated for C4H6BrN3O 191.98; found 192.0.

[00520] Step 3. A solution of (5-bromo-1-methyl-1,2,4-triazol-3-yl)methanol (6.0 g) and HBr in AcOH (144.0 mL) was stirred overnight at 80 °C. The mixture was neutralized to pH 9 with saturated NaHCO 3 (aq.). The resulting mixture was extracted with EtOAc (3 x 60 mL). The combined organic layers were washed with brine (3 x 100 mL), dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. This afforded 5-bromo-3-(bromomethyl)-1-methyl-1,2,4-triazole (6 g, crude) as a white solid. LCMS (ESI): m/z [M+H] calcd for C 4 H 5 Br 2 N 3 253.89; found 253.8.

[00521] Step 4. To a stirred mixture of 5-bromo-3-(bromomethyl)-1-methyl-1,2,4-triazole (6.0 g, 23.54 mmol) and tert-butyl 2-[(diphenylmethylidene)amino]acetate (6.95 g, 23.54 mmol) in toluene (42 mL) and DCM (18.0 mL) was added (2R,4R,5S)-1-(anthracen-9-ylmethyl)-5-ethenyl-2-[(S)-(prop-2-en-1-yloxy)(quinolin-4-yl)methyl]-1-azabicyclo[2.2.2]octan-1-ium bromide (1.43 g, 2.35 mmol) in portions at 0 °C under an argon atmosphere. The resulting mixture was stirred and KOH (60 mL) in H2O was added. The resulting mixture was stirred for 24 h at -10 °C under an argon atmosphere. The desired product could be detected by LCMS. The reaction was quenched with NH4Cl (aq.) at 0 °C. The resulting mixture was extracted with EtOAc (3 × 100 mL). The combined organic layers were washed with brine (1 × 200 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC to afford tert-butyl (2S)-3-(5-bromo-1-methyl-1,2,4-triazol-3-yl)-2-[(diphenylmethylidene)amino]propanoate (5 g, 38.6% yield) as a yellow oil. LCMS (ESI): m/z [M+H] calculated for C21H21BrN4O2 469.12; found 469.1.

[00522] Step 5. To a stirred solution of tert-butyl (2S)-3-(5-bromo-1-methyl-1,2,4-triazol-3-yl)-2-[(diphenylmethylidene)amino]propanoate (5.0 g, 10.65 mmol) in DCM (50.0 mL) was added TFA (25.0 mL) dropwise at 0 °C under an argon atmosphere. The resulting mixture was stirred for 16 h at room temperature under an argon atmosphere. The resulting mixture was concentrated under reduced pressure to give (2S)-2-amino-3-(5-bromo-1-methyl-1,2,4-triazol-3-yl)propanoic acid (6 g, crude) as a brown oil. LCMS (ESI): m/z [M+H] calculated for C6H9BrN4O2 249.00; found 249.0.

[00523] Step 6. To a stirred solution of (2S)-2-amino-3-(5-bromo-1-methyl-1,2,4-triazol-3-yl)propanoic acid (6.0 g, 24.09 mmol) in THF (36.0 mL) was added NaHCO3 (10.14 g, 120.69 mmol), Boc2O (7.89 g, 36.14 mmol) in portions at 0 °C under an argon atmosphere. The resulting mixture was stirred for 16 h at room temperature. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure. The mixture was purified by reversed-phase chromatography to afford (2S)-3-(5-bromo-1-methyl-1,2,4-triazol-3-yl)-2-[(tert-butoxycarbonyl)amino]propanoic acid (3 g, 33.9% yield) as a white solid. LCMS (ESI): m/z [M+H] calculated for C11H17BrN4O4 349.05; found 349.0.

[00524] Step 7. To a stirred solution of 2-[[(2M)-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indol-3-yl]methyl]-2-methylpropyl (3S)-1,2-diazinane-3-carboxylate (1.0 g, 1.65 mmol) in DMF (10.0 mL) was added DIPEA (4.28 g, 33.08 mmol), (2S)-3-(5-bromo-1-methyl-1,2,4-triazol-3-yl)-2-[(tert-butoxycarbonyl)amino]propanoic acid (0.69 g, 1.98 mmol) and HATU (0.75 g, 1.99 mmol) portionwise at 0 °C. The resulting mixture was stirred for 2 h at 20 °C under an argon atmosphere. The desired product could be detected by LCMS. The resulting mixture was quenched with H2O (100 mL) and extracted with EtOAc (3 × 30 mL). The combined organic layers were washed with brine (3 × 100 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase chromatography to give 2-[[(2M)-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indol-3-yl]methyl]-2-methylpropyl(3S)-1-[(2S)-3-(5-bromo-1-methyl-1,2,4-triazol-3-yl)-2-[(tert-butoxycarbonyl)amino]propanoyl]-1,2-diazinane-3-carboxylate (800 mg, 46.5% yield) as a light yellow solid. LCMS (ESI): m/z [M+H] calcd for C45H64BBrN8O8 935.42; found 935.2.

[00525] Step 8. To a stirred solution of 2-[[(2M)-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indol-3-yl]methyl]-2-methylpropyl K3PO4 (0.45 g, 2.12 mmol), XPhos (122.26 mg, 0.27 mmol), English: XPhos Pd G3 (0.22 g, 0.27 mmol) and H2O (2.0 mL) at room temperature. The resulting mixture was stirred for 3 h at 75 °C under an argon atmosphere. The desired product could be detected by LCMS. The resulting mixture was extracted with EtOAc (3 × 100 mL). The combined organic layers were washed with brine (3 × 60 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase chromatography to afford tert-butyl((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-21,10,10-trimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H,21H-8-oxa-1(5,3)-indola-6(1,3)-pyridazin-2(5,3)-triazolacycloundecaphane-4-yl)carbamate (400 mg, 56.8% yield) as a light yellow solid. LCMS (ESI): m/z [M+H] calcd for C39H52N8O6 729.41; found 729.3.

[00526] Step 9. To a solution of tert-butyl((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-21,10,10-trimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H,21H-8-oxa-1(5,3)-indola-6(1,3)-pyridazin-2(5,3)-triazolacycloundecaphane-4-yl)carbamate (400.0 mg, 0.56 mmol) in DCM (1 mL) was added TFA (0.5 mL). The reaction was stirred for 1 h at room temperature under an argon atmosphere. After concentration, the mixture was neutralized to pH 8 with saturated NaHCO3 (aq., 20 mL). The mixture was extracted with DCM (3 × 20 mL). The organic layers were dried over Na2SO4 and concentrated to afford (63S,4S,Z)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-21,10,10-trimethyl-61,62,63,64,65,66-hexahydro-11H,21H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(5,3)-triazolacycloundecaphane-5,7-dione (500 mg, crude) as a light yellow solid. ESI-MS m/z = 629.3 [M+H]+; MWcalculated:628.3. LCMS (ESI): m/z [M+H] calculated for C34H44N8O4 629.36; found 629.3.

[00527] Step 10. To a stirred solution of (63S,4S,Z)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-21,10,10-trimethyl-61,62,63,64,65,66-hexahydro-11H,21H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(5,3)-triazolacycloundecaphane-5,7-dione (170.0 mg, 0.27 mmol) and (R)-2-(((1-benzhydrylazetidin-3-yl)oxy)methyl)-3-methylbutanoic acid (114.68 mg, 0.32 mmol) in DMF (5 mL) was added DIPEA (698.86 mg, 5.41 mmol) and HATU (123.36 mg, 0.32 mmol) were added dropwise at 0 °C under an air atmosphere. The resulting mixture was stirred for 2 h at 0 °C. The resulting mixture was diluted with 25 mL of H2O. The resulting mixture was extracted with EtOAc (3 × 25 mL). The combined organic layers were washed with brine (3 × 25 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to give (2R)-2-(((1-benzhydrylazetidin-3-yl)oxy)methyl)-N-((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-21,10,10-trimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H,21H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(5,3)-triazolacycloundecaphane-4-yl)-3-methylbutanamide (180 mg, crude) as an off-white oil. LCMS (ESI): m/z [M+H] calculated for C56H69N9O6 964.54; found 964.4.

[00528] Step 11. To a stirred solution of (2R)-2-(((1-benzhydrylazetidin-3-yl)oxy)methyl)-N-((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-21,10,10-trimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H,21H-8-oxa-1(5,3)-indola-6(1,3)-pyridazin-2(5,3)-triazolacycloundecaphane-4-yl)-3-methylbutanamide (180.0 mg, 0.19 mmol) and Pd/C (90.0 mg, 0.85 mmol) in MeOH (10 mL) was added Boc2O (81.48 mg, 0.37 mmol) at room temperature under a hydrogen atmosphere. The resulting mixture was stirred at room temperature overnight. The resulting mixture was filtered, the filter cake was washed with MeOH (3x10 mL). The filtrate was concentrated under reduced pressure to afford tert-butyl 3-((2R)-2-(((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-21,10,10-trimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H,21H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(5,3)-triazolacycloundecaphane-4-yl)carbamoyl)-3-methylbutoxy)azetidine-1-carboxylate (80 mg, 47.7% yield) as an off-white solid. LCMS (ESI): m/z [M+H] calcd for C48H67N9O8 898.52; found 898.4.

[00529] Step 12. To a stirred solution of tert-butyl 3-((2R)-2-(((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-21,10,10-trimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H,21H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(5,3)-triazolacycloundecaphane-4-yl)carbamoyl)-3-methylbutoxy)azetidine-1-carboxylate in DCM (2 mL) was added TFA (1.0 mL) dropwise at 0 C under an atmosphere of air. The resulting mixture was stirred for 1 h at 0 °C. The resulting mixture was concentrated under reduced pressure to give (2R)-2-((azetidin-3-yloxy)methyl)-N-((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-21,10,10-trimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H,21H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(5,3)-triazolacycloundecaphane-4-yl)-3-methylbutanamide (85 mg, crude) as a yellow-green oil.

[00530] Step 13. A stirred solution of (2R)-2-((azetidin-3-yloxy)methyl)-N-((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-21,10,10-trimethyl- and acid 4-(dimethylamino)-4-methylpent-2-inoic acid (38.90 mg, 0.25 mmol) in DMF (2 mL) was added DIPEA (518.27 mg, 4.01 mmol) and COMU (51.52 mg, 0.12 mmol) in portions at 0 C. The reaction mixture was stirred under an air atmosphere for 2 h. The crude product (150 mg) was purified by reversed-phase chromatography to give (2R)-2-(((1-(4-(dimethylamino)-4-methylpent-2-inoyl)azetidin-3-yl)oxy)methyl)-N-((63S,4S,Z)- 11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-21,10,10-trimethyl-5,7-dioxo-61,62,63,64,65,66- hexahydro-11H,21H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(5,3)-triazolacycloundephane-4-yl)-3-methylbutanamide (15.3 mg, 16.3% yield) as a solid off-white.1H NMR (400 MHz, DMSO-d6) δ 8.77 (dd, J = 4.8, 1.7 Hz, 1H), 8.15 (d, J = 1.7 Hz, 1H), 8.07 (d, J = 8.1 Hz, 1H), 7.85 - 7.78 (m, 1H), 7.70 (d, J = 8.6 Hz, 1H), 7.58 - 7.48 (m, 2H), 5.82 (s, 1H), 4.95 (d, J = 11.7 Hz, 1H), 4.41 - 4.30 (m, 5H), 4.30 (d, J = 8.2 Hz, 2H), 4.25 (d, J = 5.6Hz, 4H), 4.10 (td, J = 17.1, 16.1, 9.1 Hz, 2H), 3.99 - 3.82 (m, 3H), 3.71 - 3.60 (m, 1H), 3.54 - 3.43 (m, 3H), 3.39 (s, 2H), 3.22 (d, J = 1.6 Hz, 1H), 2.92 (d, J = 13.6 Hz, 1H), 2.86 - 2.77 (m, 2H), 2.45 (s, 6H), 2.37 (q, J = 7.7 Hz, 1H), 2.17 (d, J = 6.6 Hz, 2H), 2.03 (d, J = 10.2 Hz, 2H), 1.78 - 1.66 (m, 3H), 1.47 (t, J = 10.9 Hz, 6H), 1.35 - 1.28 (m, 12H), 0.32 (s, 3H). LCMS (ESI): m/z [M+H] calculated for C51H70N10O7 935.55; found 935.3.EXAMPLE A692. SYNTHESIS OF (3S)-1-(4-(DIMETHYLAMINO)-4-METHYLPENT-2-INOYL)-N- ((2S)-1 -(((63S,4S)-11 -ETHYL-12-(2-((S)-1 -METHOXYETHYL)PYRIDIN-3-YL)-1 0,10-DIMETHYL- 5,7-DIOXO-61 ,62,63,64,65,66-HEXA-HYDRO-11H-8-OXA-2(5,2)-OXAZOLA-1 (5,3)- INDOLA-6(1,3)-PYRDAZINECYCLOUNDECAPHANE-4-YL)AMINO)-3-METHYL-1-OXOBUTAN-2- IL)-N-METHYLPYRROLIDINE-3-CARBOXAMIDE

[00531] Step 1. To a solution of 1,3-oxazol-2-ylmethanol (5.0 g, 50.46 mmol) in THF (75 mL) were added imidazole (8.59 mg, 0.13 mmol) and TBSCl (11.41 mg, 0.08 mmol) at 0 °C. The resulting solution was stirred for 5 h and then concentrated under reduced pressure. The crude material was purified by silica gel column chromatography to afford 2-[[(tert-butyldimethylsilyl)oxy]methyl]-1,3-oxazole (10 g, 92.8% yield) as a colorless oil. LCMS (ESI): m/z [M+H] calcd C10H19NO2Si 214.13; found 214.3.

[00532] Step 2. To a solution of 2-[[(tert-butyldimethylsilyl)oxy]methyl]-1,3-oxazole (10.0 g, 46.87 mmol) in THF (150.0 mL, 1851.45 mmol) at -78 °C was added n-BuLi (22.4 mL, 56.25 mmol) over 10 min and stirred for 30 min at -78 °C under an argon atmosphere. Then, a solution of Br2 (3.6 mL, 70.31 mmol) in THF (10 mL) was added over 10 min to the solution at -78 °C. The resulting solution was slowly warmed to room temperature and stirred for 2 h. The resulting mixture was diluted with NH4Cl/H2O (100 mL) and extracted with EtOAc (3 × 100 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography to afford 5-bromo-2-[[(tert-butyldimethylsilyl)oxy]methyl]-1,3-oxazole (5.3 g, 38.6% yield) as a yellow oil. LCMS (ESI): m/z [M+H] calcd for C10H18BrNO2Si 292.04; found 292.0.

[00533] Step 3. To a solution of 5-bromo-2-[[(tert-butyldimethylsilyl)oxy]methyl]-1,3-oxazole (4.0 g, 13.69 mmol) in DCM (60.0 mL) was added PBr3 (7.41 g, 27.37 mmol) at 0 °C under an argon atmosphere. The resulting solution was stirred for 4 h and then diluted with NaHCO3/H2O (30 mL). The mixture was extracted with EtOAc (3 x 40 mL). The organic layers were concentrated under reduced pressure and purified by silica gel column chromatography to afford 5-bromo-2-(bromomethyl)-1,3-oxazole (2.5 g, 75.7% yield) as a yellow oil. LCMS (ESI): m/z [M+H] calcd for C4H3Br2NO 239.87; found 241.9.

[00534] Step 4. A mixture of 5-bromo-2-(bromomethyl)-1,3-oxazole (9.0 g, 37.36 mmol), Cat: 200132-54-3 (2.26 g, 3.74 mmol), DCM (45.0 mL), toluene (90.0 mL), KOH (20.96 g, 373.63 mmol), H2O (42 mL), and tert-butyl 2-[(diphenylmethylidene)amino]acetate (13.24 g, 44.82 mmol) at 0 C was stirred for 4 h and then diluted with H2O (30 mL). The mixture was extracted with DCM (3 x 40 mL). The organic layers were concentrated under reduced pressure and purified by reversed-phase column chromatography to afford tert-butyl (2S)-3-(5-bromo-1,3-oxazol-2-yl)-2-[(diphenylmethylidene)amino]propanoate (4.8 g, 28.2% yield) as a yellow solid. LCMS (ESI): m/z [M+H] calculated for C23H23BrN2O3 455.10; found 457.1.

[00535] Step 5. A mixture of tert-butyl (2S)-3-(5-bromo-1,3-oxazol-2-yl)-2-[(diphenylmethylidene)amino]propanoate (1.20 g, 2.64 mmol), DCM (10.0 mL, 157.30 mmol), and TFA (5.0 mL, 67.32 mmol) at 0 °C was stirred for 2 h and then concentrated under reduced pressure to afford (S)-3-(5-bromooxazol-2-yl)-2-((2,2,2-trifluoroacetyl)-14-azaneyl)propanoic acid (0.5 g, 81.3% yield) as a yellow solid. LCMS (ESI): m/z [M+H] calcd for C6H7BrN2O3 234.97; found 237.0.

[00536] Step 6. A mixture of (S)-3-(5-bromooxazol-2-yl)-2-((2,2,2-trifluoroacetyl)-14-azaneyl)propanoic acid (500.0 mg, 2.13 mmol), Boc2O (928.56 mg, 4.26 mmol), dioxane (2.50 mL), H2O (2.50 mL), and NaHCO3 (714.84 mg, 8.51 mmol) at 0 C was stirred for 3 h. The resulting solution was purified by reversed-phase column chromatography to afford (2S)-3-(5-bromo-1,3-oxazol-2-yl)-2-[(tert-butoxycarbonyl)amino]propanoic acid (0.65 g, 91.1% yield) as a yellow solid. LCMS (ESI): m/z [M+H] calculated for C11H15BrN2O5 335.02; found 334.8.

[00537] Step 7. To a solution of (2S)-3-(5-bromo-1,3-oxazol-2-yl)-2-[(tert-butoxycarbonyl)amino]propanoic acid (500.0 mg, 1.49 mmol) and 3-[(2M)-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indol-3-yl]-2,2-dimethylpropan-1-ol (808.16 mg, 1.64 mmol) in DMF (5.0 mL) and H2O(1.0 mL) K3PO4 (791.67 mg, 3.73 mmol) and Pd(dppf)Cl2 (109.16 mg, 0.15 mmol). The resulting mixture was stirred for 2 h at 70 °C under an argon atmosphere. The mixture was purified by reversed-phase column chromatography to give (2 S)-2-[(tert-butoxycarbonyl)amino]-3-[5-[(2M)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-5-yl]-1,3-oxazol-2-yl]propanoic acid (600 mg, 64.79% yield) as a light brown solid. LCMS (ESI): m/z [M+H] calcd for C34H44N4O7 621.33; found 621.3.

[00538] Step 8. To a stirred mixture of methyl (3S)-1,2-diazinane-3-carboxylate (627.10 mg, 4.350 mmol) and (2S)-2-[(tert-butoxycarbonyl)amino]-3-[5-[(2M)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-5-yl]-1,3-oxazol-2-yl]propanoic acid (900.0 mg, 1.45 mmol) in DCM (10.0 mL) were added HATU (661.54 mg, 1.74 mmol) and DIPEA (3747.71 mg, 29.00 mmol) at 0 °C. The resulting mixture was stirred for 2 h. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure and the crude material was purified by silica gel column chromatography to afford methyl (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[5-[(2M)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-5-yl]-1,3-oxazol-2-yl]propanoyl]-1,2-diazinane-3-carboxylate (900 mg, 83.11%) as a yellow brown solid. LCMS (ESI): m/z [M+H] calcd for C40H54N6O8 747.41; found 747.2.

[00539] Step 9. To a stirred mixture of methyl (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[5-[(2M)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-5-yl]-1,3-oxazol-2-yl]propanoyl]-1,2-diazinane-3-carboxylate (2000.0 mg, 2.68 mmol) in THF (18 mL) and H2O (6.0 mL) was added LiOH.H2O (337.10 mg, 8.03 mmol) at 0 °C. The resulting mixture was stirred for 2 h. The desired product can be detected by LCMS. The reaction was quenched with H2O at 0 °C and adjusted to pH 6 with 1 N HCl solution. The resulting mixture was extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with brine (1 × 10 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to give (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[5-[(2M)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-5-yl]-1,3-oxazol-2-yl]propanoyl]-1,2-diazinane-3-carboxylic acid (1300 mg, 66.2% yield) as a yellow solid. LCMS (ESI): m/z [M+H] calculated for C39H52N6O8 733.39; found 733.3.

[00540] Step 10. To a stirred mixture of (3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[5-[(2M)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-5-yl]-1,3-oxazol-2-yl]propanoyl]-1,2-diazinane-3-carboxylic acid (1.2 g, 1.64 mmol) and DIPEA (8.5 g, 65.50 mmol) in DCM (120.0 mL) were added HOBT (1.8 g, 13.10 mmol) and EDCI (7.8 g, 40.93 mmol) at 0 °C. The resulting mixture was stirred for 2 h. The desired product could be detected by LCMS. The mixture was concentrated under reduced pressure and purified by silica gel column chromatography to give tert-butyl((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(5,2)-oxazole-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)carbamate (660 mg, 56.4% yield) as a yellow brown solid. LCMS (ESI): m/z [M+H] calcd for C39H50N6O7 715.38; found 715.3.

[00541] Step 11. A mixture of tert-butyl((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(5,2)-oxazole-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)carbamate (20.0 mg, 0.028 mmol) and TFA (3.0 mL) in DCM (6.0 mL) at 0 C was stirred for 2 h. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure to give (63S,4S)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro- 11H-8-oxa-2(5,2)-oxazole-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-5,7-dione (160 mg, crude) as a green-yellow solid. LCMS (ESI): m/z [M+H] calcd for C34H42N6O5 615.33; found 615.2.

[00542] Step 12. To a stirred mixture of (63S,4S)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(5,2)-oxazole-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-5,7-dione (180.0 mg, 0.293 mmol) and (2S)-2-[(tert-butoxycarbonyl)methyl)amino]-3-methylbutanoic acid (135.45 mg, 0.59 mmol) in DMF (2.0 mL) were added HATU (133.60 mg, 0.35 mmol) and DIPEA (756.86 mg, 5.86 mmol) at 0 °C. The resulting mixture was stirred for 2 h. The desired product could be detected by LCMS. The mixture was purified by reversed-phase column chromatography to give tert-butyl ((2S)-1-(((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo- 61,62,63,64,65,66-hexahydro-11H-8-oxa-2(5,2)-oxazole-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamate (160 mg, 66% yield) as a yellow brown solid. LCMS (ESI): m/z [M+H] calcd for C45H61N7O8 828.47; found 828.4.

[00543] Step 13. To a stirred mixture of tert-butyl ((2S)-1-(((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo- 61,62,63,64,65,66-hexahydro-11H-8-oxa-2(5,2)-oxazola-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamate (160.0 mg, 0.19 mmol) in DCM (2.0 ml) TFA (1.0 ml) was added at 0°C. The resulting mixture was stirred for 2 h. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure to give (2S)-N-((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(5,2)-oxazole-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)-3-methyl-2-(methylamino)butanamide (130 mg, crude) as a yellow solid. LCMS (ESI): m/z [M+H] calcd for C40H53N7O6 728.41; found 728.5.

[00544] Step 14. To a stirred mixture of acid ((2S)-N-((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo- 61,62,63,64,65,66-hexahydro-11H-8-oxa-2(5,2)-oxazole-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-4-yl)-3-methyl-2-(methylamino)butanamide (130.0 mg, 0.18 mmol) and acid (3S)-1-[4-(dimethylamino)-4-methylpent-2-inoyl]pyrrolidine-3-carboxylic acid (180.25 mg, 0.72 mmol) in DMF (2.0 mL) were added DIPEA (461.64 mg, 3.57 mmol) and HATU (135.81 mg, 0.36 mmol) at 0 C. The resulting mixture was stirred for 2 h. The desired product could be detected by LCMS. The mixture was purified by reversed-phase column chromatography to give (3S)-1-(4-(dimethylamino)-4-methylpent-2-ynoyl)-N-((2S)-1-(((63S,4S)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexa- hydro-11H-8-oxa-2(5,2)-oxazole-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)-N-methylpyrrolidine-3-carboxamide (55.3 mg, 31.3% yield) as a solid. 1H NMR (400 MHz, DMSO-d6) δ 8.77 (dd, J = 4.8, 1.7 Hz, 1H), 8.09 - 8.00 (m, 1H), 7.92 (s, 1H), 7.88 - 7.80 (m, 1H), 7.62 (d, J = 8.7 Hz, 1H), 7.59 - 7.49 (m, 2H), 7.39 - 7.30 (m, 1H), 5.69 (p, J = 8.8 Hz, 1H), 5.44 (d, J = 12.1 Hz, 1H), 4.67 (d, J = 10.7 Hz, 1H), 4.30 - 4.15 (m, 3H), 3.99 (dt, J = 13.2, 6.4 Hz, 3H), 3.89 - 3.79 (m, 1H), 3.62 (ddd, J = 30.5, 18.6, 11.4 Hz, 5H), 3.39 (dd, J = 9.4, 3.8 Hz, 2H), 3.21 - 3.13 (m, 1H), 3.08 (d, J = 15.0 Hz, 3H), 2.99 - 2.74 (m, 6H), 2.26 - 2.18 (m, 5H), 2.16 (s, 2H), 2.14 - 1.94 (m, 3H), 1.86 - 1.68 (m, 2H), 1.57 (q, J = 9.2, 5.8 Hz, 1H), 1.44 - 1.27 (m, 9H), 0.94 (d, J = 6.6 Hz, 4H), 0.89 (dd, J = 6.5, 2.5 Hz, 2H), 0.80 (d, J = 6.3 Hz, 2H), 0.77 - 0.69 (m, 4H), 0.58 (d, J = 20.4 Hz, 3H). LCMS (ESI): m/z [M+H] calculated for C53H71N9O8 962.55; found 962.5. EXAMPLE A675. SYNTHESIS OF (2R)-2-(((1-(4-(DIMETHYLAMINO)-4-METHYLPENT-2- INOYL)AZETIDIN-3-YL)OXY)METHYL)-N-((63S,4S,Z)-11 -ETHYL-12-(2-((S)-1 - METHOXYETHYL)PYRIDIN-3-IL)-10,10-DIMETHYL-5,7-DIOXO-61,62,63,64,65,66-HEXA-HYDRO- 11H-8-OXA-2(2,4)-OXAZOLA-1(5,3)-INDOLE-6(1,3)-PYIDAZINECYCLOUNDECAPHANE-4-YL)-3-METHYLBUTANAMIDE

[00545] Step 1. A mixture of 2-bromo-4-(ethoxycarbonyl)-1,3-oxazol-5-ylium (6.83 g, 31.19 mmol), EtOH (100.0 mL), and NaBH4 (4.72 g, 124.76 mmol) at 0 °C was stirred for 6 h at 0 °C under an air atmosphere. The reaction was quenched with H2O at 0 °C. The resulting mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to afford (2-bromo-1,3-oxazol-4-yl)methanol (4.122 g, 74.3% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C4H4BrNO2 177.95; found 178.0.

[00546] Step 2. A mixture of (2-bromo-1,3-oxazol-4-yl)methanol (4.30 g, 24.16 mmol), DCM (50 mL), and phosphorus tribromide (9809.39 mg, 36.24 mmol) at 0 °C was stirred overnight at 0 °C under an air atmosphere. The reaction was quenched by the addition of NaHCO3 (aq.) at 0 °C. The resulting mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to afford 2-bromo-4-(bromomethyl)-1,3-oxazole (3.28 g, 56.4% yield) as a liquid. LCMS (ESI): m/z [M+H] calculated for C4H3Br2NO 239.87; found 239.9.

[00547] Step 3. A mixture of 2-bromo-4-(bromomethyl)-1,3-oxazole (3280.0 mg, 13.62 mmol), KOH (9M, 10 mL), 30 mL of toluene/DCM (7/3) mixture, and tert-butyl 2-[(diphenylmethylidene)amino]acetate (5228.74 mg, 17.70 mmol) at -16 °C was stirred overnight under an air atmosphere. The resulting mixture was extracted with EtOAc (3 x 50 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to afford tert-butyl tert-butyl (2S)-3-(2-bromo-1,3-oxazol-4-yl)-2-[(diphenylmethylidene)amino]propanoate (8.33 g, 80.6% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C23H23BrN2O3 455.10; found 455.1.

[00548] Step 4. A mixture of tert-butyl (2S)-3-(2-bromo-1,3-oxazol-4-yl)-2-[(diphenylmethylidene)amino]propanoate (4100.0 mg, 9.0 mmol) and citric acid (1 N) (40.0 mL, 0.21 mmol), in THF (40 mL) at room temperature was stirred overnight under an atmosphere of air. The reaction was quenched by the addition of HCl (aq.) (100 mL) at 0 °C. The aqueous layer was extracted with EtOAc (3 x 100 mL). K2CO3 (aq.) (200 mL) was added to the resulting mixture and extracted with EtOAc (3 x 100 mL). The organic layer was concentrated under reduced pressure to afford tert-butyl (2S)-2-amino-3-(2-bromo-1,3-oxazol-4-yl)propanoate (1730 mg, 66% yield) as a dark yellow solid. LCMS (ESI): m/z [M+H] calculated for C10H15BrN2O3 291.03; found 291.0.

[00549] Step 5. A mixture of tert-butyl (2S)-2-amino-3-(2-bromo-1,3-oxazol-4-yl)propanoate (1780.0 mg, 6.11 mmol), TFA (10.0 mL), and DCM (10.0 mL) at 0 °C was stirred overnight under an atmosphere of air. The resulting mixture was concentrated under reduced pressure to afford (2S)-2-amino-3-(2-bromo-1,3-oxazol-4-yl)propanoic acid (1250 mg, 87% yield) as a dark yellow solid. LCMS (ESI): m/z [M+H] calcd for C6H7BrN2O3 234.97; found 234.9.

[00550] Step 6. A mixture of di-tert-butyl dicarbonate (4178.58 mg, 19.15 mmol), THF (10 mL), H2O (10 mL), (2S)-2-amino-3-(2-bromo-1,3-oxazol-4-yl)propanoic acid (1500.0 mg, 6.38 mmol), and NaHCO3 (3216.74 mg, 38.29 mmol) at room temperature was stirred overnight under an atmosphere of air. The reaction was quenched with H2O at room temperature. The resulting mixture was concentrated under reduced pressure. The resulting mixture was extracted with EtOAc (3 x 100 mL). The aqueous layer was acidified to pH 6 with 1 M HCl (aq.). The resulting mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to afford (2S)-3-(2-bromo-1,3-oxazol-4-yl)-2-[(tert-butoxycarbonyl)amino]propanoic acid (920 mg, 43.0% yield) as an oil. LCMS (ESI): m/z [M+H] calculated for C11H15BrN2O5 335.02; found 335.0.

[00551] Step 7. A mixture of 3-(2-bromo-1,3-oxazol-4-yl)-2-[(tert-butoxycarbonyl)amino]propanoic acid (850.0 mg, 2.54 mmol), methyl 1,2-diazinane-3-carboxylate (1.88 g, 13.04 mmol), DIPEA (1966.68 mg, 15.22 mmol), DCM (30.0 mL), and HATU (1446.48 mg, 3.80 mmol) at 0 °C was stirred for 3 h under an air atmosphere. The resulting mixture was extracted with DCM (3 x 50 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The resulting mixture was purified by reverse flash chromatography to afford methyl 1-[3-(2-bromo-1,3-oxazol-4-yl)-2-[(tert-butoxycarbonyl)amino]propanoyl]-1,2-diazinane-3-carboxylate (610 mg, 52.1% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C17H25BrN4O6 461.10; found 461.0.

[00552] Step 8. A mixture of methyl 1-[3-(2-bromo-1,3-oxazol-4-yl)-2-[(tert-butoxycarbonyl)amino]propanoyl]-1,2-diazinane-3-carboxylate (570.0 mg, 1.24 mmol), LiOH (2.0 mL, 1 M aq.), and THF (2.0 mL) at 0 °C was stirred for 3 h under an air atmosphere. The resulting mixture was concentrated under reduced pressure. The resulting mixture was extracted with EtOAc (3 × 50 mL). The combined aqueous layers were acidified to pH 5 with 1N HCl (aq.). The aqueous phase was extracted with EtOAc (3 × 50 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to afford 1-[3-(2-bromo-1,3-oxazol-4-yl)-2-[(tert-butoxycarbonyl)amino]propanoyl]-1,2-diazinane-3-carboxylic acid (500 mg, 90.5% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C16H23BrN4O6 447.09; found 446.8.

[00553] Step 9. A mixture of 1-[3-(2-bromo-1,3-oxazol-4-yl)-2-[(tert-butoxycarbonyl)amino]propanoyl]-1,2-diazinan-3-carboxylic acid (450.0 mg, 1.01 mmol), 3-[(2M)-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indol-3-yl]-2,2-dimethylpropan-1-ol (743.19 mg, 1.51 mmol), DMAP (24.58 mg, 0.20 mmol), DCM (15.0 mL) and DCC (311.37 mg, 1.51 mmol) at 0 °C was Stirred for 3 h under an air atmosphere. The resulting mixture was extracted with EtOAc (3 × 20 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC to give 3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl(S)-1-((S)-3-(2-bromooxazol-4-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate (330 mg, 35.6% yield) as a white solid. LCMS (ESI): m/z [M+H] calcd for C45H62BBrN6O9 921.39; found 921.4.

[00554] Step 10. A mixture of 3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)- 2,2-dimethylpropyl(S)-1-((S)-3-(2-bromo-oxazol-4-yl)-2-((tert-butoxycarbonyl)amino)propanoyl) hexahydropyridazine-3-carboxylate(290.0 mg, 0.33 mmol), K3PO4 (206.64 mg, 0.97 mmol), X-Phos (30.94 mg, 0.07 mmol), XPhos Pd G3 (54.93 mg, 0.07 mmol), dioxane (5 mL) and H2O (1.0 mL) at 70 °C was stirred for 4 h under an argon atmosphere. The resulting mixture was extracted with EtOAc (3 × 20 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC to afford tert-butyl((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(2,4)-oxazole-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)carbamate (130 mg, 56.0% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C39H50N6O7 715.38; found 715.3.

[00555] Step 11. A mixture of tert-butyl ((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66- hexahydro-11H-8-oxa-2(2,4)-oxazole-1(5,3)-indole-6(1,3)-pyridazinecycloundephane-4-yl)carbamate (120.0 mg), DCM (2.0 mL) and TFA (0.2 mL) at room temperature was stirred for 6 h under an air atmosphere. The resulting mixture was concentrated under reduced pressure to give (63S,4S,Z)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(2,4)-oxazole-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-5,7-dione (90 mg, 87.2% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C34H42N6O5 615.33; found 615.3.

[00556] Step 12. A mixture of (63S,4S,Z)-4-amino-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-61,62,63,64,65,66-hexahydro-11H-8-oxa- 2(2,4)-oxazole-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-5,7-dione (200.0 mg, 0.33 mmol), (2R)-2-([[1-(diphenylmethyl)azetidine-3-yl]oxy]methyl)-3-methylbutanoic acid (172.49 mg, 0.49 mmol), DIPEA (420.48 mg, 3.25 mmol), DMF (3.0 mL) and HATU (148.44 mg, 0.39 mmol) at 0 C was stirred for 3 h under an air atmosphere. The resulting mixture was extracted with EtOAc (3 × 20 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC to give (2R)-2-(((1-benzhydrylazetidin-3-yl)oxy)methyl)-N-((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(2,4)-oxazole-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)-3-methylbutanamide (154 mg, 77.0% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C56H67N7O7 950.52; found 950.6.

[00557] Step 13. A mixture of (2R)-2-(((1-benzhydrylazetidine-3-yl)oxy)methyl)-N-((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl- 5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(2,4)-oxazole-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-4-yl)-3-methylbutanamide (240.0 mg, 0.25 mmol), (Boc)2O (165.37 mg, 0.76 mmol), MeOH (5.0 mL) and Pd(OH)2 (72.0 mg, 0.51 mmol) at room temperature was stirred overnight under an atmosphere of H2. The resulting mixture was filtered, the filter cake was washed with MeOH (3 x 5 mL). The filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC to afford tert-butyl 3-((2R)-2-(((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(2,4)-oxazole-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)carbamoyl)-3-methylbutoxy)azetidine-1-carboxylate (150 mg, 67.2% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C48H65N7O9 884.49; found 884.2.

[00558] Step 14. A mixture of tert-butyl 3-((2R)-2-(((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo- and TFA (0.40 mL) at 0°C was stirred for 3 h under an air atmosphere. The resulting mixture was concentrated under reduced pressure to give (2R)-2-((azetidin-3-yloxy)methyl)-N-((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(2,4)-oxazole-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)-3-methylbutanamide (120 mg, 90.2% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C43H57N7O7 784.44; found 784.2.

[00559] Step 15. A mixture of (2R)-2-((azetidin-3-yloxy)methyl)-N-((63S,4S,Z)-11-ethyl-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7- dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(2,4)-oxazole-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-4-yl)-3-methylbutanamide (130.0 mg, 0.17 mmol), 4- Sodium (dimethylamino)-4-methylpent-2-inoate (44.07 mg, 0.25 mmol), DMF (3.0 mL), DIPEA (64.29 mg, 0.50 mmol) and COMU (106.46 mg, 0.25 mmol) at 0 °C was stirred for 3 h under an air atmosphere. The resulting mixture was extracted with EtOAc (3 × 20 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by reverse phase chromatography to give (2R)-2-(((1-(4-(dimethylamino)-4-methylpent-2-inoyl)azetidin-3-yl)oxy)methyl)-N-((63S,4S,Z)-11-ethyl-12-(2-((S)-1- methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(2,4)-oxazole-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-4-yl)-3-methylbutanamide (25 mg, 16.4% yield) as a solid. 1H NMR (400 MHz, DMSO-d6) δ 8.77 (dd, J = 4.8, 1.8 Hz, 1H), 8.55 (s, 1H), 8.14 (dd, J = 8.3, 2.9 Hz, 1H), 7.82 (d, J = 7.6 Hz, 1H), 7.76 - 7.65 (m, 3H) 4.18 - 4.04 (m, 2H), 3.94 (dd, J = 32.9, 7.8 Hz, 1H), 3.77 - 3.63 (m, 2H), 3.57 (s, 1H), 3.49 (s, 2H), 3.21 (s, 3H), 2.90 (d, J = 14.6 Hz, 1H), 2.87 - 2.79 (m, 1H), 2.72 (td, J = 15.5, 14.6, 3.1 Hz, 2H), 2.46 (s, 1H), 2.43 - 2.26 (m, 6H), 2.11 - 1.99 (m, 1H), 1.82 - 1.66 (m, 2H), 1.56 - 1.37 (m, 11H), 0.89 (dt, J = 12.3, 7.7 Hz, 12H), 0.35 (s, 3H). LCMS (ESI): m/z [M+H] calcd for C51H68N8O8 921.52; found 921.5.EXAMPLE A607. SYNTHESIS OF (2R)-2-(((1-(4-(DIMETHYLAMINO)-4-METHYLPENT-2- INOYL)AZETIDIN-3-YL)OXY)METHYL)-N-((23S,63S,4S)-11 -ETI-12-(2-((S)-1 - METHOXYETHYL)PYRIDIN-3-IL)-10,10-DIMETHYL-5,7-DIOXO-61,62,63,64,65,66-HEXA-HYDRO- 11H-8-OXA-1(5,3)-INDOLE-6(1,3)-PYRDAZINE-2(1,3)-PIPERIDINECYCLOUNDECAPHANE-4-YL)-3-METHYLBUTANAMIDE

[00560] Step 1. A mixture of Zn (44.18 g, 675.41 mmol) and I2 (8.58 g, 33.77 mmol) in DMF (120 mL) was stirred for 30 min at 50 °C under an argon atmosphere, followed by the addition of methyl (2R)-2-[(tert-butoxycarbonyl)amino]-3-iodopropanoate (72.24 g, 219.51 mmol) in DMF (200 mL). The reaction mixture was stirred at 50 °C for 2 h under an argon atmosphere. Next, a mixture of 2,6-dibromo-pyridine (40 g, 168.85 mmol) and Pd(PPh3)4 (39.02 g, 33.77 mmol) in DMF (200 mL) was added. The resulting mixture was stirred at 75 °C for 2 h, then cooled to room temperature and extracted with EtOAc (1 L × 3). The combined organic layers were washed with H 2 O (1 L × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give methyl (2S)-3-(6-bromopyridin-2-yl)-2-[(tert-butoxycarbony)amino]propanoate (41 g, 67% yield) as an oil. LCMS (ESI): m/z [M+H] calcd for C 14 H 19 BrN 2 O 4 358.1; found 359.1.

[00561] Step 2. To a solution of 3-[(2M)-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethylpropan-1-ol (45.0 g, 82.35 mmol) in dioxane (400 mL) and H2O (80 mL), potassium carbonate (28.45 g, 205.88 mmol), methyl (2S)-3-(6-bromopyridin-2-yl)-2-[(tert-butoxycarbonyl)amino]propanoate (35.5 g, 98.8 mmol) were added. Pd(dtbpf)Cl2 (5.37 g, 8.24 mmol) at room temperature. The reaction mixture was stirred at 70 °C for 2 h under a nitrogen atmosphere. The resulting mixture was extracted with EtOAc (500 mL × 3). The combined organic layers were washed with H2O (300 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography to give (S)-2-((tert-butoxycarbonyl)amino)-3-(6-(3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-pyridin-2-yl)propanoate (48 g, 83% yield) as a solid. LCMS (ESI): m/z [M+H] calcd C37H45F3N4O6 698.3; found 699.4.

[00562] Step 3. A solution of (S)-2-((tert-butoxycarbonyl)amino)-3-(6-(3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)pyridin-2-yl)propanoate (52 g, 74.42 mmol) in THF (520 mL) was added LiOH (74.41 mL, 223.23 mmol) at 0 °C. The reaction mixture was stirred at room temperature for 3 h. The resulting mixture was acidified to pH 5 with HCl (aq.) and extracted with EtOAc (1 L x 3). The combined organic layers were washed with H2O (1 L × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give (S)-2-((tert-butoxycarbonyl)amino)-3-(6-(3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)pyridin-2-yl)propanoic acid (50 g, 98% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C36H43F3N4O6 684.3; found 685.1.

[00563] Step 4. To a solution of (S)-2-((tert-butoxycarbonyl)amino)-3-(6-(3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)pyridin-2-yl)propanoic acid (55 g, 80.32 mmol) in DCM (600 mL) were added DIPEA (415.23 g, 3212.82 mmol) and HATU (45.81 g, 120.48 mmol) at 0 °C. The reaction mixture was stirred at room temperature for 12 h and then quenched with H2O. The resulting mixture was extracted with EtOAc (1 L x 3). The combined organic layers were washed with H2O (1 L), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give methyl 1-(S)-2-((tert-butoxycarbonyl)amino)-3-(6-(3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indole-5-pyridin-2-yl)propanoyl)hexahydropyridazine-3-carboxylate (63 g, 96% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C42H53F3N6O7 810.4; found 811.3.

[00564] Step 5. A solution of 1-((S)-2-((tert-butoxycarbonyl)amino)-3-(6-(3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)pyridin-2-yl)propanoyl)hexahydropyridazine-3-carboxylate (50 g, 61.66 mmol) in THF (500 mL) and 3M LiOH (61.66 mL, 184.980 mmol) at 0 °C was stirred at room temperature for 3 h, then acidified to pH 5 with HCl (aq.). The resulting mixture was extracted with EtOAc (800 mL × 3). The combined organic layers were washed with H2O (800 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give 1-(S)-2-((tert-butoxycarbonyl)amino)-3-(6-(3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)pyridin-2-yl)propanoyl)hexahydropyridazine-3-carboxylic acid (48 g, 97% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C41H51F3N6O7 796.3; found 797.1.

[00565] Step 6. To a solution of 1-((S)-2-((tert-butoxycarbonyl)amino)-3-(6-(3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H)-indol-5-yl)pyridin-2-yl)propanoyl)hexahydropyridazine-3-carboxylic acid (50 g, 62.74 mmol) in DCM (10 L) at 0 °C were added DIPEA (243.28 g, 1882.32 mmol), EDCI (360.84 g, 1882.32 mmol), and HOBT (84.78 g, 627.44 mmol). The reaction mixture was stirred at room temperature for 3 h, quenched with H2O, and concentrated under reduced pressure. The residue was extracted with EtOAc (2 L × 3). The combined organic layers were washed with H2O (2 L × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to afford tert-butyl ((4S)-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(2,6)-pyridinecycloundecaphane-4-yl)carbamate (43.6 g, 89% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C41H49F3N6O6 778.3; found 779.3.

[00566] Step 7. To a solution of tert-butyl((4S)-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(2,6)-pyridinecycloundecaphane-4-yl)carbamate (300 mg) in DCM (10 mL), TFA (3 mL) was added at 0 C. The reaction mixture was stirred at room temperature for 1 h. The resulting mixture was diluted with toluene (10 mL) and concentrated under reduced pressure three times to give (4S)-4-amino-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(2,6)-pyridinecycloundecaphane-5,7-dione (280 mg, crude) as oil. LCMS (ESI): m/z [M+H] calcd for C36H41F3N6O4 679.2; found 678.3.

[00567] Step 8. To a solution of (4S)-4-amino-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro- 11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(2,6)-pyridinecycloundecaphane-5,7-dione (140 mg, 0.21 mmol) in MeCN (2 mL), were added DIPEA (266.58 mg, 2.06 mmol), N-(4-(tert-butoxycarbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carbonyl)-N-methyl-L-valine (127.94 mg, 0.31 mmol) and CIP (114.68 mg, 0.41 mmol) were obtained at 0 °C. The reaction mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated under reduced pressure. The residue was extracted with EtOAc (10 mL × 3). The combined organic layers were washed with H2O (10 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography to give tert-butyl 9-(((2S)-1-(((63S,4S)-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(2,6)-pyridinecycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)-1-oxa-4,9-diazaspiro[5,5]undecane-4-carboxylate (170 mg, 76% yield) as solid. LCMS (ESI): m/z [M+H] calculated for C56H74F3N9O9 1073.5; found 1074.6.

[00568] Step 9. To a solution of tert-butyl 9-(((2S)-1-(((63S,4S)-12-(2-((S)-1-methoxyethyl) pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine- 2(2,6)-pyridinecycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl) (methyl)carbamoyl)- 1-oxa-4,9-diazaspiro [5.5] undecane-4-carboxylate (160 mg, 0.15 mmol) in DCM (5 mL) at 0 °C, TFA (1.5 mL) was added dropwise. The reaction mixture was stirred at 0 °C for 1 h. The resulting mixture was diluted with toluene (10 mL) and concentrated under reduced pressure three times to give N-((2S)-1-(((63S,4S)-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(2,6)-pyridinecycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)-N-methyl-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxamide (150 mg, crude) as oil. LCMS (ESI): m/z [M+H] calculated for C51H65F3N9O7 973.5; found 974.4.

[00569] Step 10. To a solution of N-((2S)-1-(((63S,4S)-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indola-6(1,3)-pyridazine-2(2,6)-pyridinecycloundecaphane-4-yl)amino)-3-methyl-1-oxobutan-2-yl)-N-methyl-1-oxa-4,9-diazaspiro[5,5]undecane-9-carboxamide (150 mg, 0.15 mmol) in DMF (3 mL), was added DIPEA (199.01 mg, 1.54 mmol), acrylic acid (16.64 mg, 0.23 mmol), and COMU (98.39 mg, 0.23 mmol) were prepared at 0 C. The reaction mixture was stirred at room temperature for 1 h and concentrated under reduced pressure. The residue was extracted with EtOAc (10 mL × 3). The combined organic layers were washed with H 2 O (10 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography and reverse phase chromatography to generate 4-acryloyl-N-((2S)-1-(((63S,4S)-12-(2-((S)-1- methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-1(5,3)-indole-6(1,3)-pyridazine-2(2,6)-pyridinecycloundecaphane- 4-yl)amino)-3-methyl-1-oxobutan-2-yl)-N-methyl-1-oxa-4,9-diazaspiro[5,5]undecane-9-carboxamide (53 mg, 33% yield) as a solid. 1H NMR (400 MHz, DMSO-d6) δ 8.79 (dd, J = 4.8, 1.9 Hz, 2H), 8.09 (d, J = 31.4 Hz, 1H), 7.96 (d, J = 8.9 Hz, 1H), 7.90 - 7.72 (m, 3H), 7.64 (t, J = 7.8 Hz, 1H), 7.56 (dd, J = 7.8, 4.7 Hz, 1H), 7.03 (s,1H), 6.86 (dd, J = 16.6, 10.4 Hz, 1H), 6.20 (d, J = 14.0 Hz, 1H), 5.73 (d, J = 12.2 Hz,2H), 5.47 (s, 1H), 5.35 (d, J = 11.8 Hz, 1H), 4.68 (s, 1H), 4.28 (d, J = 12.5 Hz, 1H),4.13 (d, J = 6.4 Hz, 1H), 3.92 (s, 1H), 3.84 - 3.64 (m, 6H), 3.59 (d, J = 14.0 Hz, 3H),3.50 (s, 3H), 3.10 (s, 5H), 3.02 (d, J = 13.3 Hz, 2H), 2.85 (d, J = 12.1 Hz, 3H), 2.08 - 1.89 (m, 2H), 1.81 (s, 1H), 1.75 - 1.51 (m, 5H), 1.39 (d, J = 6.1 Hz, 4H), 1.24 (s, 0H), 0.91 - 0.66 (m, 10H), 0.53 (s, 3H). LCMS (ESI): m/z [M+H] calculated for C54H68F3N9O8 1027.5; found 1028.1.EXAMPLE A590. SUMMARY OF (2R)-2-(((1-(4-(DIMETHYLAMINO)-4-METHYLPENT-2-INOYL)AZETIDIN-3-YL)OXY)METHYL)- N-((63S,4S,Z)-12-(2-((S)-1-METHOXYETHYL)PYRIDIN-3-IL)-10,10-DIMETHYL-5,7-DIOXO- 11-(2,2,2-TRIFLUOROETHYL)-61,62,63,64,65,66-HEXA-HYDRO-11H-8-OXA-2(5,3)-THIADIAZOLA-1(5,3)-INDOLE-6(1,3)-PYRDAZINECYCLOUNDECAPHANE-4-YL)-3-METHYLBUTANAMIDE

[00570] Step 1. To a mixture of ethyl 2-ethoxy-2-iminoacetate (25.0 g, 172.23 mmol) and EtOH (250.0 mL) at 0 °C was added ammonium chloride (9.21 g, 172.23 mmol) in portions and then stirred for 4 h at room temperature under an argon atmosphere. The resulting mixture was concentrated under reduced pressure and washed with Et2O (3 × 200 mL). The organic layers were combined and concentrated under reduced pressure. This afforded ethyl 2-amino-2-iminoacetate hydrochloride (20 g, crude) as a light yellow solid. LCMS (ESI): m/z [M+H] calcd for C4H8N2O2 117.07; found 116.9.

[00571] Step 2. To a mixture of ethyl 2-amino-2-iminoacetate hydrochloride (13.30 g, 87.17 mmol), H2O (50.0 mL), and Et2O (100.0 mL) at 0 °C was added sodium hypochlorite pentahydrate (7.79 g, 104.60 mmol) dropwise. The resulting mixture was stirred for 3 h under an argon atmosphere. The mixture was extracted with Et2O (3x 200 mL). The resulting solution was washed with brine (3x 100 mL). The organic phase was dried over anhydrous Na2SO4 and concentrated under reduced pressure to afford (Z)-2-amino-2-(chloroimino)ethyl acetate (7 g, crude) as a light yellow solid. LCMS (ESI): m/z [M+H] calculated for C4H7ClN2O2 151.03; found 150.8.

[00572] Step 3. To a solution of ethyl (Z)-2-amino-2-(chloroimino)acetate (8.40 g, 55.792 mmol) and MeOH (130.0 mL) at 0 °C was added potassium thiocyanate (5.42 g, 55.79 mmol) in portions. The resulting mixture was stirred for 4 h at room temperature under an argon atmosphere. The reaction was quenched with H2O/Ice. The mixture was extracted with EtOAc (5 x 100 mL). The organic phase was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to afford ethyl 5-amino-1,2,4-thiadiazole-3-carboxylate (2.3 g, 23.8% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C5H7N3O2S 174.03; found 173.8.

[00573] Step 4. To a solution of ethyl 5-amino-1,2,4-thiadiazole-3-carboxylate (5.80 g, 33.49 mmol), MeCN (90.0 mL), and CuBr2 (11.22 g, 50.23 mmol) at 0 °C was added 2-methyl-2-propylnitrite (6.91 g, 66.98 mmol) dropwise under an argon atmosphere. The mixture was stirred for 30 min. The mixture was then stirred for 4 h at 50 °C. The mixture was cooled to 0 °C and quenched with H2O/ice. The mixture was extracted with EtOAc (3x100 mL). The organic phase was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by silica gel chromatography to give ethyl 5-bromo-1,2,4-thiadiazole-3-carboxylate (6.2 g, 78.1% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C5H5BrN2O2S 236.93; found 237.1.

[00574] Step 5. To a solution of (S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropan-1-ol (16.60 g, 33.24 mmol), DCM (170.0 mL), and imidazole (5.66 g, 83.10 mmol) at 0 °C was added tert-butyl-chlorodiphenylsilane (11.88 g, 43.21 mmol) dropwise. The resulting mixture was stirred for 3 h at room temperature under an argon atmosphere. The reaction was quenched with H2O/ice. The mixture was extracted with EtOAc (3x 200 mL). The organic layer was washed with brine (3x100 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give (S)-5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indole (22 g, 89.7% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C39H44BrF3N2O2Si 737.24; found 737.0.

[00575] Step 6. To a solution of (S)-5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indole (28.0 g, 37.95 mmol), toluene (270.0 mL), KOAc (9.31 g, 94.88 mmol), and bis(pinacolato)diboron (19.27 g, 75.90 mmol) at 0 °C was added Pd(dppf)Cl2.CH2Cl2 (6.18 g, 7.59 mmol) in portions. The resulting mixture was stirred for 3 h at 90 °C under an argon atmosphere. The mixture was cooled to room temperature and quenched with H2O/ice. The mixture was extracted with EtOAc (3 × 200 mL). The organic phase was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain (S)-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-(1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2-trifluoroethyl)-1H-indole (28.2 g, 94.7% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C45H56BF3N2O4Si 785.41; found 785.4.

[00576] Step 7. To a solution of (S)-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-(1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5- tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2-trifluoroethyl)-1H)-indole (19.60 g, 24.97 mmol), 1,4-dioxane (200 mL), H2O (40 mL), ethyl 5-bromo-1, 2, 4-thiadiazol-3-carboxylate (5.92 g, 24.97 mmol) and K3PO4 (13.25 g, 62.43 mmol) at 0 °C Pd(dtbpf)Cl2 was added (1.63 g, 2.50 mmol) in portions. The resulting mixture was stirred for 1.5 h at 75 °C under an argon atmosphere. The mixture was cooled to 0 °C and quenched with H 2 O/ice and extracted with EtOAc (3 × 200 mL). The organic phase was dried over anhydrous Na 2 SO 4 and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give ethyl (S)-5-(3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)-1,2,4-thiadiazole-3-carboxylate (14 g, 68.8% yield) as a yellow solid. LCMS (ESI): m/z [M+H] calculated for C44H49F3N4O4SSi 815.33; found 815.2.

[00577] Step 8. To a solution of ethyl (S)-5-(3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)-1,2,4-thiadiazole-3-carboxylate (13.60 g, 16.69 mmol) and EtOH (140.0 mL) at 0 °C was added NaBH 4 (3.16 g, 83.43 mmol) in portions. The resulting mixture was stirred for 3 h and then quenched with H 2 O/ice. The resulting mixture was washed with brine (3 x 100 mL) and extracted with EtOAc (3 x 200 mL). The combined organic layers were dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give (S)-5-(3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)-1,2,4-thiadiazol-3-ol (9.7 g, 75.2% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C42H47F3N4O3SSi 773.32; found 773.3.

[00578] Step 9. To a solution of (S)-5-(3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)-1,2,4-thiadiazol-3-ol (9.70 g, 12.55 mmol), DCM (100.0 mL), and CBr4 (8.32 g, 25.10 mmol) at 0 °C was added PPh3 (6.58 g, 25.10 mmol) in DCM (20.0 mL) dropwise. The resulting mixture was stirred for 2 h under an argon atmosphere and then quenched with H2O/ice. The mixture was extracted with DCM (3 × 200 mL). The organic phase was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by reverse flash chromatography to give (S)-3-(bromomethyl)-5-(3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)-1,2,4-thiadiazole (9.5 g, 90.6% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C42H46BrF3N4O2SSi 835.23; found 834.9.

[00579] Step 10. To a stirred solution of (S)-3-(bromomethyl)-5-(3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)-1,2,4-thiadiazole (9.40 g, 11.25 mmol), toluene (84.0 mL), DCM (36.0 mL), , tert-butyl 2-[(diphenylmethylidene)amino]acetate (3.32 g, 11.25 mmol), and O-allyl-N-(9-anthracenylmethyl)cinconidinium bromide (0.68 g, 1.13 mmol) at 0 °C was added 9 M NaCl of aqueous KOH (94.0 mL) dropwise. The resulting mixture was stirred overnight under an argon atmosphere. The mixture was extracted with EtOAc (3 × 200 mL), and the organic phase was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give tert-butyl (S)-3-(5-(3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)-1,2,4-thiadiazol-3-yl)-2-((diphenylmethylene)amino)propanoate (9 g, 76.2% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C61H66F3N5O4SSi 1050.46; found 1050.8.

[00580] Step 11. To a solution of tert-butyl (S)-3-(5-(3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)-1,2,4-thiadiazol-3-yl)-2-((diphenylmethylene)amino)propanoate (8.0 g, 7.62 mmol) and DCM (40.0 mL) at 0 °C was added TFA (40.0 mL) dropwise. The resulting mixture was stirred overnight at room temperature and then concentrated under reduced pressure. The residue was basified to pH 8 with NaHCO 3 . The mixture was extracted with EtOAc (3 × 200 mL). The organic phase was concentrated under reduced pressure. The residue was purified by reverse flash chromatography to give (S)-2-amino-3-(5-(3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)-1,2,4-thiadiazol-3-yl)propanoic acid (5 g, 79.1% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C44H50F3N5O4SSi 830.34; found 830.2.

[00581] Step 12. To a solution of (S)-2-amino-3-(5-(3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)-1,2,4-thiadiazol-3-yl)propanoic acid (4.70 g, 5.66 mmol), DCM (50.0 mL), and Et3N (2.86 g, 28.31 mmol) at 0 °C was added (Boc)2O (1.36 g, 6.23 mmol) dropwise. The resulting mixture was stirred for 3 h at room temperature under an argon atmosphere and then concentrated under reduced pressure and purified by reverse flash chromatography to give (S)-2-((tert-butoxycarbonyl)amino)-3-(5-(3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)-1,2,4-thiadiazol-3-yl)propanoic acid (5 g, 94.9% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C49H58F3N5O6SSi 930.39; found 930.3.

[00582] Step 13. To a mixture of (S)-2-((tert-butoxycarbonyl)amino)-3-(5-(3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)-1,2,4-thiadiazol-3-yl)propanoic acid (5.30 g, 5.70 mmol), DMF (60.0 mL), methyl 1,2-diazinane-3-carboxylate (1.64 g, 11.40 mmol), and DIPEA (22.09 g, 170.94 mmol) at 0 °C was added HATU (2.82 g, 7.41 mmol) in DMF (5 mL) was added dropwise. The resulting mixture was stirred for 3 h at room temperature under an argon atmosphere. The reaction was then quenched with H2O/ice. The mixture was extracted with EtOAc (3 × 100 mL), and the organic phase was washed with brine (3 × 100 mL). The resulting mixture was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to afford methyl (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(5-(3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)-1,2,4-thiadiazol-3-yl)propanoyl)hexahydropyridazine-3-carboxylate (5.6 g) as a solid. LCMS (ESI): m/z [M+H] calcd for C55H68F3N7O7SSi 1056.47; found 1056.2.

[00583] Step 14. A mixture of methyl (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(5-(3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)-1,2,4-thiadiazol-3-yl)propanoyl)hexahydropyridazine-3-carboxylate (5.60 g, 5.30 mmol) and TBAF in THF (56.0 mL) was stirred overnight at 40 C under an argon atmosphere. The reaction was quenched with NH4Cl (aq.). The mixture was extracted with EtOAc (3 × 100 mL), and the organic phase was concentrated under reduced pressure. The residue was purified by reverse flash chromatography to give (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(5-(3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)-1,2,4-thiadiazol-3-yl)propanoyl)hexahydropyridazine-3-carboxylic acid (4.1 g, 96.2% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C38H48F3N7O7S 804.34; found 804.3.

[00584] Step 15. To a solution of (S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(5-(3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)-1,2,4-thiadiazol-3-yl)propanoyl)hexahydropyridazine-3-carboxylic acid (4.0 g, 5.0 mmol) and DCM (450.0 mL) at 0 °C were added DIPEA (51.45 g, 398.08 mmol), HOBt (6.72 g, 49.76 mmol), and EDCI (57.23 g, 298.55 mmol) in portions. The resulting mixture was stirred for 16 h at room temperature under an argon atmosphere. The reaction was quenched with H2O/ice and extracted with EtOAc (3 × 30 mL). The organic phase was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to afford tert-butyl((63S,4S,Z)-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(5,3)-thiadiazole-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)carbamate (1.7 g, 43.5% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C38H46F3N7O6S 786.33; found 786.3.

[00585] Step 16. To a solution of ((63S,4S,Z)-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(5,3)-thiadiazole-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)carbamate (300.0 mg, 0.38 mmol) and DCM (2.0 mL) at 0 °C was added TFA (1.0 mL) dropwise. The resulting mixture was stirred for 2 h at room temperature and then concentrated under reduced pressure. The residue was basified to pH 8 with saturated NaHCO3 (aq.). The mixture was extracted with EtOAc (3 × 20 mL). The organic phase was concentrated under reduced pressure to give (63S,4S,Z)-4-amino-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(5,3)-thiadiazole-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-5,7-dione (270 mg, crude) as a solid. LCMS (ESI): m/z [M+H] calculated for C33H38F3N7O4S 686.27; found 686.1.

[00586] Step 17. To a solution of (63S,4S,Z)-4-amino-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(5,3)-thiadiazole-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-5,7-dione (160.0 mg, 0.23 mmol), (R)-2-(((1-benzhydrylazetidin-3-yl)oxy)methyl)-3-methylbutanoic acid (123.70 mg, 0.35 mmol) and DMF (2.0 mL) at 0 °C were added DIPEA (603.09 mg, 4.660 mmol) and COMU (119.91 mg, 0.28 mmol) in DMF (0.5 mL). The resulting mixture was stirred for 2 h at room temperature under an argon atmosphere. The reaction was quenched with H2O/ice and extracted with EtOAc (3 × 20 mL). The organic phase was washed with brine (3 × 10 mL) and concentrated under reduced pressure. The residue was purified by Prep-TLC to give (2R)-2-(((1-benzhydrylazetidin-3-yl)oxy)methyl)-N-((63S,4S,Z)-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(5,3)-thiadiazole-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)-3-methylbutanamide (160 mg, 67.2% yield) as a solid. LCMS (ESI): m/z [M+H] calcd for C55H63F3N8O6S 1021.46; found 1021.4.

[00587] Step 18. To a solution of (2R)-2-(((1-benzhydrylazetidine-3-yl)oxy)methyl)-N-((63S,4S,Z)-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)- 10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(5,3)-thiadiazole-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-4-yl)-3-methylbutanamide (160.0 mg, 0.16 mmol) and MeOH (5.0 mL) at 0 C was added (Boc)2O (85.49 mg, 0.39 mmol) dropwise followed by Pd/C (320.0 mg) portionwise. The resulting mixture was stirred overnight at room temperature under a hydrogen atmosphere. The resulting mixture was filtered, and the filter cake was washed with EtOAc (3 × 20 mL). The filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC to afford tert-butyl 3-((2R)-2-(((63S,4S,Z)-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(5,3)-thiadiazole-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)carbamoyl)-3-methylbutoxy)azetidine-1-carboxylate (80 mg, 53.5% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C47H61F3N8O8S 955.44; found 955.2.

[00588] Step 19. To a solution of tert-butyl 3-((2R)-2-(((63S,4S,Z)-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(5,3)-thiadiazole-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)carbamoyl)-3-methylbutoxy)azetidine-1-carboxylate (120.0 mg, 0.13 mmol) and DCM (0.80 mL) at 0 °C was added TFA (0.4 mL) dropwise and the resulting mixture was stirred for 2 h at room temperature. The mixture was basified to pH 8 with saturated NaHCO3 (aq.). The mixture was extracted with EtOAc (3 × 10 mL) and concentrated under reduced pressure. The residue was purified by reverse flash chromatography to give (2R)-2-((azetidin-3-yloxy)methyl)-N-((63S,4S,Z)-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(5,3)-thiadiazole-1(5,3)-indola-6(1,3)-pyridazinecycloundecaphane-4-yl)-3-methylbutanamide (40 mg, 37.2% yield) as a solid. LCMS (ESI): m/z [M+H] calculated for C42H53F3N8O6S 855.38; found 855.3.

[00589] Step 20. To a solution of acid (2R)-2-((azetidin-3-yloxy)methyl)-N-((63S,4S,Z)-12-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7- dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(5,3)-thiadiazole-1(5,3)-indole-6(1,3)-pyridazinecycloundecaphane-4-yl)-3-methylbutanamide (32.0 mg, 0.037 mmol),4-(dimethylamino)-4-methylpent-2-inoic acid (11.62 mg, 0.074 mmol) and DMF (0.50 mL) at 0 °C were added DIPEA (193.49 mg, 1.48 mmol) and COMU (19.23 mg, 0.044 mmol) in DMF (0.1 mL) dropwise. The resulting mixture was stirred for 2 h at room temperature under an argon atmosphere. The reaction was quenched with H2O/ice and extracted with EtOAc (3 × 20 mL). The organic phase was washed with brine (3 × 10 mL) and concentrated under reduced pressure. The crude product (60 mg) was purified by reversed-phase chromatography to give (2R)-2-(((1-(4-(dimethylamino)-4-methylpent-2-inoyl)azetidin-3-yl)oxy)methyl)-N-((63S,4S,Z)-12-(2-((S)-1- methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)- 61,62,63,64,65,66-hexahydro-11H-8-oxa-2(5,3)-thiadiazole-1(5,3)-indole-6(1,3)- pyridazinecycloundecaphane-4-yl)-3-methylbutanamide (11.7 mg, 30.9% yield) as a solid. 1H NMR (400 MHz, DMSO-d6) δ 8.80 (dd, J = 4.7, 1.8 Hz, 1H), 8.58 (s, 1H), 8.30 (d, J = 8.9 Hz, 1H), 7.92 (d, J = 8.6 Hz, 1H), 7,847.69 (m, 2H), 7.56 (dd, J = 7.8, 4.8 Hz, 1H), 5.78 (t, J = 8.6 Hz, 2H), 5.10 (d, J = 12.1 Hz, 1H), 4.91 (dd, J = 16.9, 8.8 Hz, 1H), 4.31 (d, J = 6.6 Hz, 6H), 4.05 (dd, J = 16.3, 6.6 Hz, 2H), 3.87 (d, J = 6.3 Hz, 1H), 3.75 (d, J = 11.3 Hz, 1H), 3.61 (d, J = 11.0 Hz, 1H), 3.53 (d, J = 9.8 Hz, 1H), 3.45 (s, 3H), 3.25 (s, 1H), 3.09 (d, J = 10.4 Hz, 1H), 3.01 (d, J = 14.5 Hz, 1H), 2.79 (s, 1H), 2.45-2.35 (m, 2H), 2.20 (d, J = 5.4 Hz, 6H), 2.15 (d, J = 12.0Hz, 1H), 1.81 (d, 2H), 1.74-1.65 (m, 1H), 1.54 (s, 1H), 1.41-1.30 (m, 9H), 1.24 (s, 1H), 0.94 (s, 3H), 0.90-0.81 (m, 5H), 0.29 (s, 3H). LCMS (ESI): m/z [M+H] calculated for C50H64F3N9O7S 992.47; found 992.5.

[00590] The following table of compounds (Table 3) was prepared using the methods mentioned above or variations thereof, as is known to those skilled in the art. TABLE 3: EXEMPLARY COMPOUNDS PREPARED BY METHODS OF THE PRESENT INVENTION Blank = not determined.

CORRESPONDING PAIR ANALYSIS

[00591] FIGS. 1A-1B compare the potency in two different cell-based assays of Formula BB compounds of the present invention (right dots) and corresponding compounds of Formula AA (left dots) in which one H is replaced by (S)Me. The y-axes represent pERK EC50 (FIG. 1A) or CTG IC50 (FIG. 1B) as measured in an H358 cell line. Assay protocols are below. Connected dots represent a matched pair that differs only between the H and (S)Me substitution. Each Formula BB compound demonstrated reduced potency in cellular assays compared to the corresponding Formula AA compound.

BIOLOGICAL TESTS POWER TEST: PERK

[00592] The objective of this assay is to measure the ability of test compounds to inhibit K-Ras in cells. Activated K-Ras induces increased phosphorylation of ERK at Threonine 202 and Tyrosine 204 (pERK). This procedure measures a decrease in cellular pERK in response to test compounds. The procedure described below in NCI-H358 cells is applicable to K-Ras G12C.

[00593] Note: This protocol can be performed substituting other cell lines to characterize inhibitors of other RAS variants, including, for example, AsPC-1 (K-Ras G12D), Capan-1 (KRas G12V), or NCI-H1355 (K-Ras G13C).

[00594] NCI-H358 cells were cultured and maintained using media and procedures recommended by the ATCC. The day prior to compound addition, cells were plated in 384-well cell culture plates (40 μL/well) and grown overnight in a 37°C, 5% CO2 incubator. Test compounds were prepared in 10-fold dilutions in DMSO, with a high concentration of 10 mM. On the day of the assay, 40 nL of the test compound was added to each well of the cell culture plate using an Echo550 liquid handler (LabCyte®). Test compound concentrations were tested in duplicate. After compound addition, cells were incubated 4 hours at 37°C, 5% CO2. Following incubation, the culture medium was removed and the cells were washed once with phosphate buffered saline.

[00595] In some experiments, the cellular pERK level was determined using the AlphaLISA SureFire Ultra p-ERK1/2 Assay Kit (PerkinElmer). Cells were lysed in 25 μL of lysis buffer, with shaking at 600 RPM at room temperature. The lysate (10 μL) was transferred to a 384-well Opti plate (PerkinElmer) and 5 μL of acceptor mix was added. After a 2-hour incubation in the dark, 5 μL of donor mix was added, the plate was sealed and incubated 2 hours at room temperature. The signal was read on an Envision plate reader (PerkinElmer) using default AlphaLISA settings. Raw data analysis was performed in Excel (Microsoft) and Prism (GraphPad). The signal was plotted vs. the decadal logarithm of compound concentration and IC50 was determined by fitting a 4-parameter sigmoidal concentration response model.

[00596] In further experiments, cellular pERK was determined by In-Cell Western. After compound treatment, cells were washed twice with 200 μL of tris-buffered saline (TBS) and fixed for 15 minutes with 150 μL of 4% paraformaldehyde in TBS. Fixed cells were washed 4 times for 5 minutes with TBS containing 0.1% Triton X-100 (TBST) and then blocked with 100 μL of Odyssey blocking buffer (LI-COR) for 60 minutes at room temperature. Primary antibody (pERK, CST-4370, Cell Signaling Technology) was diluted 1:200 in blocking buffer and 50 μL was added to each well and incubated overnight at 4°C. Cells were washed 4 times for 5 minutes with TBST. Secondary antibody (rabbit IR-800CW, LI-COR, diluted 1:800) and DRAQ5 DNA stain (LI-COR, diluted 1:2000) were added and incubated 1-2 hours at room temperature. Cells were washed 4 times for 5 minutes with TBST. Plates were scanned on a Li-COR Odyssey CLx Imager. Raw data analysis was performed in Excel (Microsoft) and Prism (GraphPad). Signal was plotted vs. decadal logarithm of compound concentration and IC50 was determined by fitting a 4-parameter sigmoidal concentration response model.

[00597] The following compounds exhibited a pERK EC50 of less than 5 uM (H358 KRAS G12C):A48,A15,A272,A174,A163,A453,A447,A279,A240,A214,A225,A136,A226,A219,A228,A21,A12,A78,A424,A219,A378,A224,A4,A53,A187,A218,A213,A314,A22 0,A208,A24,A9,A126,A345,A46,A203,A210,A184,A469,A366,A113,A328,A693, A639,A364,A100,A249,A486,A307,A347,A33,A210,A192,A285,A468,A185,A61 2,A109,A284,A200,A2,A6,A606,A325,A139,A496,A393,A561,A125,A494,A547, A215,A258,A195,A259,A212,A637,A53,A63,A68,A178,A189,A205,A78,A254,A 690,A563,A14,A19,A92,A576,A278,A331,A42,A67,A209,A350,A562,A652,A70 3,A623,A191,A241,A199,A193,A478,A251,A177,A222,A23,A59,A26,A211,A10 6,A279,A120,A7,A134,A521,A116,A467,A694,A729,A151,A110,A277,A340,A2 21,A723,A13,A442,A611,A50,A190,A553,A696,A211,A303,A613,A37,A146,A6 66,A688,A216,A390,A548,A238,A160,A183,A164,A451,A481,A524,A1,A186,A 37,A635,A71,A269,A289,A489,A400,A731,A497,A568,A274,A253,A471,A720, A241,A179,A180,A426,A117,A363,A716,A423,A217,A708,A227,A3,A12,A8,A3 81,A84,A408,A85,A171,A263,A473,A258,A564,A118,A103,A565,A641,A655,A 47,A11,A392,A169,A487,A640,A206,A449,A358,A192,A148,A4,A41,A5,A18,A3 01,A10,A65,A554,A159,A264,A99,A79,A142,A143,A25,A98,A80,A101,A730,A2 12,A359,A61,A441,A283,A413,A717,A145,A182,A62,A181,A233,A232,A634,A 495,A34,A251,A539,A632,A54,A327,A37,A196,A607,A645,A35,A214,A225,A6 38,A40,A52,A268,A448,A575,A176,A593,A15,A17,A94,A170,A713,A93,A402,A 64,A261,A399,A422,A214,A225,A625,A31,A119,A135,A281,A676,A709,A81,A 32,A633,A39,A646,A662,A124,A732,A320,A81,A187,A354,A45,A570,A165,A6 6,A20,A455,A431,A270,A250,A457,A153,A404,A710,A541,A127,A373,A369,A 557,A349,A598,A618,A60,A636,A499,A87,A156,A680,A477,A406,A330,A202, A535,A617,A737,A201,A302,A722,A209,A374,A631,A29,A555,A420,A380,A11 1,A306,A173,A628,A672,A51,A167,A588,A512,A194,A282,A412,A701,A583,A 396,A678,A649,A27,A204,A626,A257,A614,A409,A172,A372,A353,A58,A728, A74,A619,A144,A183,A538,A445,A531,A360,A361,A459,A536,A344,A267,A57 4,A677,A530,A415,A30,A73,A152,A490,A702,A714,A483,A567,A43,A310,A31 9,A86,A321,A656,A739,A115,A130,A155,A608,A648,A168,A485,A738,A129,A 650,A715,A488,A147,A121,A470,A115,A133,A510,A421,A309,A335,A387,A38 6,A734,A95,A430,A604,A458,A592,A384,A664,A197,A725,A89,A83,A586,A62 2,A305,A498,A668,A427,A630,A158,A644,A735,A70,A683,A352,A341,A719,A 674,A70,A44,A501,A438,A698,A377,A417,A154,A433,A104,A184,A603,A280, A712,A237,A105,A394,A605,A517,A704,A566,A77,A356,A454,A600,A643,A11 2,A569,A529,A247,A463,A437,A718,A472,A461,A558,A48,A671,A395,A670,A 681,A687,A382,A82,A686,A342,A436,A296,A16,A545,A533,A416,A149,A207, A371,A596,A675,A132,A419,A56,A579,A733,A573,A707,A597,A697,A75,A653 ,A362,A615,A332,A69,A162,A128,A432,A654,A22,A397,A526,A582,A418,A91, A260,A97,A191,A55,A581,A375,A522,A108,A367,A610,A552,A571,A57,A543, A661,A138,A196,A246,A337,A446,A265,A96,A509,A123,A627,A651,A682,A15 7,A572,A624,A691,A532,A462,A580,A695,A186,A316,A540,A590,A665,A244, A166,A587,A629,A595,A518,A519,A131,A502,A726,A452,A141,A181,A262,A3 38,A155,A389,A124,A275,A414,A546,A679,A425,A669,A28,A520,A88,A131,A 589,A621,A182,A297,A594,A283,A194,A250,A336,A706,A252,A440,A107,A72 4,A525,A388,A175,A300,A333,A659,A346,A150,A476,A368,A528,A503,A504, A505,A684,A76,A736,A551,A383,A491,A492,A493,A410,A316,A295,A559,A51 1,A38,A140,A663,A334,A700,A692,A348,A584,A513,A657,A328,A515,A317,A 135,A660,A351,A544,A281,A685,A602,A556,A385,A326,A464,A465,A403,A13 3,A299,A667,A255,A334,A256,A585,A642,A133,A443,A435,A560,A444,A439, A324,A120,A407,A527,A245,A370,A537,A247,A474,A475,A705,A323,A112,A2 98,A609,A673,A292,A599,A132,A145,A266,A601,A466,A549,A379,A727,A167, A711,A75,A76,A121,A357,A620,A316,A479,A290,A339,A322,A376,A456,A391 ,A291,A550,A343,A721,A689,A411,A578,A616,A534,A365,A658,A699,A577,A 647,A591,A542,A279,A294.

DETERMINATION OF CELL VIABILITY IN RAS MUTANT CANCER CELL LINES PROTOCOL: CELLTITER-GLO® CELL VIABILITY ASSAY

[00598] Note - The following protocol describes a procedure for monitoring the cell viability of K-Ras mutant cancer cell lines in response to a compound of the invention. Other RAS isoforms may be employed, although the number of cells to be seeded will vary based on the cell line used.

[00599] The objective of this cellular assay was to determine the effects of test compounds on the proliferation of three human cancer cell lines (NCI-H358 (K-Ras G12C), AsPC-1 (K-Ras G12D), and Capan-1 (K-Ras G12V)) over a 5-day treatment period by quantifying the amount of ATP present at endpoint using CellTiter-Glo® 2.0 Reagent (Promega).

[00600] Cells were seeded at 250 cells/well in 40 μL of growth medium in 384-well assay plates and incubated overnight in a humidified atmosphere of 5% CO2 at 37°C. On the day of the assay, 10 mM stock solutions of test compounds were first diluted into 3 mM solutions with 100% DMSO. Well-mixed compound solutions (15 μL) were transferred to the next wells containing 30 μL of 100% DMSO and repeated until a 3-fold serial dilution of concentration 9 was made (initial assay concentration of 10 μM). Test compounds (132.5 nL) were dispensed directly into the cell-containing assay plates. Plates were shaken for 15 seconds at 300 rpm, centrifuged, and incubated in a humidified atmosphere of 5% CO2 at 37°C for 5 days. On day 5, assay plates and their contents were equilibrated at room temperature for approximately 30 minutes. CellTiter-Glo® 2.0 reagent (25 μL) was added and plate contents were mixed for 2 minutes on an orbital shaker prior to incubation at room temperature for 10 minutes. Luminescence was measured using the PerkinElmer Enspire. Data were normalized by the following: (Sample signal/DMSO mean)*100. Data were fit using a four-parameter logistic fit.

DISRUPTION OF THE INTERACTION OF THE RAS B-RAF BINDING DOMAIN (BRAFRBD) WITH K-RAS BY COMPOUNDS OF THE INVENTION (ALSO CALLED FRET ASSAY OR MOA ASSAY)

[00601] Note - The following protocol describes a procedure for monitoring the disruption of K-Ras G12C (GMP-PNP) binding to BRAFRBD by a compound of the invention. This protocol may also be performed by substituting other Ras proteins or nucleotides.

[00602] The objective of this biochemical assay was to measure the ability of test compounds to facilitate ternary complex formation between a nucleotide-loaded K-Ras isoform and Cyclophilin A; the resulting ternary complex disrupts binding to a BRAFRBD construct, inhibiting K-Ras signaling through a RAF effector. Data are reported as IC50 values.

[00603] In assay buffer containing 25 mM HEPES pH7.3, 0.002% Tween20, 0.1% BSA, 100 mM NaCl 5 mM MgCl2, untagged Cyclophilin A, His6-K-Ras-GMPPNP, and GST-BRAFRBD were combined in a 384-well assay plate at final concentrations of 25 μM, 12.5 nM, and 50 nM, respectively. Compound was present in plate wells as a 10-point, 3-fold dilution series starting at a final concentration of 30 μM. After incubation at 25°C for 3 hours, a mixture of Anti-His Eu-W1024 and anti-GST allophycocyanin was then added to the assay sample wells at final concentrations of 10 nM and 50 nM, respectively, and the reaction incubated for an additional 1.5 hours. The TR-FRET signal was read on a microplate reader (Ex 320 nm, Em 665/615 nm). Compounds that facilitate the disruption of a K-Ras:RAF complex were identified as those that elicit a decrease in the TR-FRET ratio relative to DMSO control wells. TABLE 4: BIOLOGICAL ASSAY DATA FOR REPRESENTATIVE COMPOUNDS OF THE PRESENT INVENTION TABLE 5. ADDITIONAL H358 CELL VIABILITY ASSAY DATA (K-RASG12C, IC50, uM):*Key:++++: IC50 > 1 uM+++: 1 uM > IC50 > 0.1 uM++: 0.1 uM > IC50 > 0.01 uM+: IC50 < 0.01 uM ADDITIONAL RAS-RAF/FRET/MOA DISRUPTION TEST DATA (IC50, UM):*Key:+++++: IC50 > 10 uM++++: 10 uM > IC50 > 1 uM+++: 1 uM > IC50 > 0.1 uM++: 0.1 uM > IC50 > 0.01 uM+: IC50 < 0.01 uMTABLE 6. KRAS G12S FRET DATA TABLE 7. FRET DATA KRAS G12D TABLE 8. KRAS G13C FRET DATA TABLE 9. FRET DATA KRAS G12V TABLE 10. FRET KRAS WT DATA TABLE 11. FRET DATA KRAS G12C TABLE 12. KRAS G13D FRET DATA TABLE 13. FRET DATA KRAS Q61H TABLE 14. FRET NRAS G12C DATA TABLE 15. FRET NRAS Q61R DATA TABLE 16. NRAS Q61K FRET DATA TABLE 17. NRAS WT FRET DATA

IN VITRO CELL PROLIFERATION PANELS

[00604] Potency for cell growth inhibition was assessed at CrownBio using standard methods. Briefly, cell lines were cultured in appropriate media and then plated on 3D methylcellulose. Cell growth inhibition was determined by CellTiter-Glo® after 5 days of culture with increasing compound concentrations. Compound potency was reported as the 50% inhibition concentration (absolute IC50). The assay was run for 7 days. On day 1, cells in 2D culture were harvested during logarithmic growth and suspended in culture media at 1x105 cells/ml. Higher or lower cell densities were used for some cell lines based on prior optimization. 3.5 ml of cell suspension was mixed with 6.5% growth media with 1% methylcellulose, resulting in a cell suspension in 0.65% methylcellulose. 90 μl of this suspension was distributed into the wells of 2 96-well plates. One plate was used for the day 0 readout and 1 plate was used for the endpoint experiment. The plates were incubated overnight at 37°C, 5% CO2. On day 2, one plate (for t0 readout) was removed and 10 μl of growth medium plus 100 μl of CellTiter-Glo® Reagent were added to each well. After mixing and a 10-minute incubation, luminescence was recorded on an EnVision Multi-Label Reader (Perkin Elmer). Compounds in DMSO were diluted in growth medium so that the maximum final compound concentration was 10 μM and 4-fold serial dilutions were performed to generate a 9-point concentration series. 10 μl of compound solution at 10 times the final concentration was added to the wells of the second plate. The plate was then incubated for 120 h at 37°C and 5% CO2. On day 7, the plates were removed, 100 μl of CellTiter-Glo® Reagent was added to each well, and after mixing and a 10-min incubation, luminescence was recorded on an EnVision Multi-Label Reader (Perkin Elmer). Data were exported to GeneData Screener and modeled with a sigmoidal concentration-response model to determine the IC50 for compound response.

[00605] Not all cell lines with a given RAS mutation may be equally sensitive to a RAS inhibitor targeting that mutation, due to differential expression of efflux transporters, varying dependencies on RAS pathway activation for growth, or other reasons. This was exemplified by the KYSE-410 cell line, which despite harboring a KRAS G12C mutation, is insensitive to the KRAS G12C (OFF) inhibitor MRTX-849 (Hallin et al., Cancer Discovery 10:54–71 (2020)) and the SW1573 cell line, which is insensitive to the KRAS G12C (OFF) inhibitor AMG510 (Canon et al., Nature 575:217–223 (2019)).TABLE 18: IC50 VALUES FOR VARIOUS CANCER CELL LINES WITH COMPOUND B*Key:low sensitivity: IC50 > 1 uMmoderately sensitive: 1 uM > IC50 > 0.1 uM very sensitive: IC50 < 0.1 uM TABLE 19: SUMMARY OF IC50 RESULTS FOR VARIOUS CANCER CELL LINES WITH VARIOUS COMPOUNDS OF THE PRESENT INVENTION (B AND M COMPOUNDS)*Key:(L) low sensitivity: IC50 > 1 uM(M) moderately sensitive: 1 uM > IC50 > 0.1 uM(V) very sensitive: IC50 < 0.1 uM

IN VIVO NSCLC K-RAS G12C XENOGRAFT MODELS COMPOUND A: METHODS:

[00606] The effects of a compound of the present invention, Compound A (H358 pERK K-Ras G12C EC50: 0.001 uM), on tumor cell growth in vivo were evaluated in the NCI-H358 KRASG12C human non-small cell lung cancer xenograft model using female BALB/c nude mice (6-8 weeks old). Mice were implanted with NCI-H358 tumor cells in 50% Matrigel (5 x 106 cells/mouse) subcutaneously in the flank. At the indicated tumor volume (dotted line, FIG. 2A), mice were randomized to treatment groups to begin administration of test articles or vehicle. Compound A was administered by oral gavage daily at a dose of 100 mg/kg. Body weight and tumor volume (using calipers) were measured twice weekly until study endpoints. The spaghetti plot (FIG. 2B) shows the change in tumor volume in individual tumors during the course of treatment.

RESULTS:

[00607] FIG. 2A shows that Compound A dosed at 100mg/kg by daily oral gavage led to tumor regression in the NCI-H358 KRASG12C xenograft model, which is a model sensitive to KRASG12C inhibition alone. The spaghetti titer plot (FIG. 2B) displaying individual tumor growth is shown alongside the tumor volume plot (FIG. 2A). Over the 28-day treatment course, Compound A led to tumor regression in all 10 animals bearing NCI-H358 KRASG12C tumors.

COMPOUND B: METHODS:

[00608] The combinatorial effect of a compound of the present invention, Compound B (H358 pERK K-Ras G12C EC50: 0.003 uM), with cobimetinib on tumor cell growth in vivo was evaluated in the NCI-H358 KRASG12C human non-small cell lung cancer xenograft model using female BALB/c nude mice (6-8 weeks old). Mice were implanted with NCI-H358 tumor cells in 50% Matrigel (5 x 106 cells/mouse) subcutaneously in the flank. At the indicated tumor volume (dotted line, FIG. 3A), mice were randomized to treatment groups to begin administration of test articles or vehicle. Compound B was administered by intermittent (twice weekly) intravenous injection at a dose of 50 mg/kg. Cobimetinib was administered by daily oral gavage at 2.5 mg/kg. The combination of Compound B and cobimetinib at their respective single-agent dose and regimen was also tested. Body weight and tumor volume (using calipers) were measured twice weekly until study endpoints. End-of-study responses in individual tumors were represented as a waterfall plot (Fig. 3B), and numbers indicate the number of tumor regressions in each group. Tumor regression is defined as a greater than 10% reduction in tumor volume at the end of the study relative to baseline.

RESULTS:

[00609] FIG. 3A shows the combination of intermittent intravenous administration of Compound B at 50 mg/kg plus daily oral administration of cobimetinib at 2.5 mg/kg led to tumor regression, while each single agent led to inhibition of tumor growth. End-of-study responses were shown as waterfall plots (FIG. 3B), which indicate that 6 out of 10 mice had tumor regression in the combination group, while no tumor regression was recorded in each single agent group.

COMPOUND C: METHODS:

[00610] The combinatorial effect of a compound of the present invention, Compound C (H358 pERK K-Ras G12C EC50: 0.007 uM), with a SHP2 inhibitor, RMC-4550, on tumor cell growth in vivo was evaluated in the NCI-H358 KRASG12C human non-small cell lung cancer xenograft model using female BALB/c nude mice (6-8 weeks old). Mice were implanted with NCI-H358 tumor cells in 50% Matrigel (5 x 106 cells/mouse) subcutaneously in the flank. At the indicated tumor volume (dotted line, FIG. 4A), mice were randomized to treatment groups to begin administration of test articles or vehicle. Compound C was administered by intravenous injection once weekly at a dose of 60 mg/kg. The SHP2 inhibitor was administered by daily oral gavage at 30 mg/kg. The combination of Compound C and SHP2 inhibitor at their respective dose and single-agent regimen was also tested. Body weight and tumor volume (using calipers) were measured twice weekly until study endpoints. End-of-study responses in individual tumors were represented as a waterfall plot (Fig. 4B), and numbers indicate the number of tumor regressions in each group. Tumor regression is defined as a greater than 10% reduction in tumor volume at the end of the study relative to baseline.

RESULTS:

[00611] In FIG. 4A, the combinatorial activity of once-weekly intravenous administration of Compound C at 60 mg/kg plus daily oral administration of SHP2 inhibitor at 30 mg/kg is shown. The combination treatment had similar antitumor activity to the single-agent SHP2 inhibitor, but the combination treatment led to 8 of 10 mice with tumor regression, while the single-agent SHP2 inhibitor led to 5 of 10 mice with tumor regressions. Single-agent Compound C administered once-weekly by intravenous injection led to inhibition of tumor growth with one tumor regression.

CELL PROLIFERATION ASSAY METHODS:

[00612] NCI-H358 cells were plated in 12-well tissue culture plates at a density of 100,000 cells/well in RPMI 1640 (10% FBS, 1% PenStrep) and cultured overnight at 37°C, 5% CO2. The next day, cells were treated with trametinib (10 nM) or a compound of the present invention, Compound D (H358 pERK K-Ras G12C EC50: 0.024 uM), (17 nM). These concentrations represent the EC50 values from a 72-hour proliferation assay using CellTiter-Glo® reagent (Promega). Additionally, cells were treated with the combination of trametinib and Compound D at the concentrations indicated above. The plate was placed in the Incucyte S3 live cell analysis system (37°C, 5% CO2) and confluence was measured by recording images at 6-hour intervals for a maximum of 40 days, or until the wells reached maximal confluence. Medium and drug were replaced at 3–4-day intervals. Data are plotted as % confluence over the course of the experiment for each single agent and respective combination (Fig. 5).

RESULTS:

[00613] As shown in FIG. 5, treatment of NCI-H358 cells with submaximal concentrations (EC50) of Compound D or MEK inhibitor results in a short period of growth inhibition, followed by proliferation. Cells reach maximal confluence within ~10 days after drug addition. The combination of the MEK inhibitor, trametinib, with Compound D resulted in complete and sustained inhibition of cell growth throughout the duration of the assay.

[00614] Although the invention has been described in conjunction with specific embodiments thereof, it will be understood that the same is capable of further modifications and this application is intended to cover all variations, uses or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as are within the known or customary practice in the art to which the invention belongs and can be applied to essential features disclosed herein.

[00615] All publications, patents and patent applications are hereby incorporated by reference in their entirety to the same extent as if each individual publication, patent or patent application were specifically and individually indicated to be incorporated by reference in its entirety.

Claims (26)

1. COMPOUND, or pharmaceutically acceptable salt thereof, characterized by having the structure of Formula If: Formula Ifwhere A is -N(H or CH3)C(O)-(CH2)- where the amino nitrogen is bonded to the carbon atom of -CH(R10)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 6-membered heteroarylene;B is -CH(R9)- where the carbon is bonded to the carbonyl carbon of NHC(O)-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;L is absent or is a ligand, wherein the ligand is the structure of Formula II:A1-(B1)f-(C1)g-(B2)h-(D1)-(B3)i-(C2)j-(B4)k-A2Formula IIwhere A1 is a bond between the ligand and B; A2 is a bond between W and the ligand; B1, B2, B3, and B4 each independently are selected from optionally substituted C1-C2 alkylene, optionally substituted C1-C3 heteroalkylene, O, S, and NRN; RN is hydrogen, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted C1-C7 heteroalkyl; C1 and C2 are each independently selected from carbonyl, thiocarbonyl, sulfonyl, or phosphoryl; f, g, h, i, j, and k are each independently 0 or 1; and D1 is optionally substituted C1-C10 alkylene, optionally substituted C2-C10 alkenylene, optionally substituted C2-C10 alkynylene, optionally substituted 3- to 14-membered heterocycloalkylene, optionally substituted 5- to 10-membered heteroarylene, optionally substituted 3- to 8-membered cycloalkylene, optionally substituted 6- to 10-membered arylene, optionally substituted C2-C10 polyethylene glycolene, or optionally substituted C1-C10 heteroalkylene, or a chemical bond linking A1-(B1)f-(C1)g-(B2)h- to -(B3)i-(C2)j-(B4)k-A2; W is a crosslinking group selected from a vinyl ketone, a vinyl sulfone, an ynone, or an alkynyl sulfone; wherein the vinyl ketone has the structure of Formula IIIa: Formula IIIawherein R16a, R16b, and R16c are independently hydrogen, -CN, halogen, or -C1-C3 alkyl optionally substituted by one or more substituents independently selected from -OH, -O-C1-C3 alkyl, -NH2, -NH(C1-C3 alkyl), -N(C1-C3 alkyl)2, or a 4- to 7-membered saturated heterocycloalkyl;the vinyl sulfone has the structure of Formula IIIc: Formula IIIcwhere R18a, R18b, and R18c are independently hydrogen, -CN, or -C1-C3 alkyl optionally substituted by one or more substituents independently selected from -OH, -O-C1-C3 alkyl, -NH2, -NH(C1-C3 alkyl), -N(C1-C3 alkyl)2, or a saturated 4- to 7-membered heterocycloalkyl;the ynone has the structure of Formula IIIb: Formula IIIwhere R17 is hydrogen, -C1-C3 alkyl optionally substituted by one or more substituents independently selected from -OH, -O-C1-C3 alkyl, -NH2, -NH(C1-C3 alkyl), -N(C1-C3 alkyl)2, or a 4- to 7-membered saturated cycloalkyl, or a 4- to 7-membered saturated heterocycloalkyl;the alkynyl sulfone has the structure of Formula IIId: Formula III wherein R19 is hydrogen, -C1-C3 alkyl optionally substituted by one or more substituents independently selected from OH, -O-C1-C3 alkyl, -NH2, -NH(C1-C3 alkyl), -N(C1-C3 alkyl)2, or a 4- to 7-membered saturated heterocycloalkyl, or a 4- to 7-membered saturated heterocycloalkyl, R1 is cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl; R2 is optionally substituted C1-C6 alkyl, C1-C6 fluoroalkyl, or optionally substituted 3- to 6-membered cycloalkyl substituted;R7 is C1-C3 alkyl;R8 is C1-C3 alkyl; andR9 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl,wherein the compound is not: (Compound A647). 2. The compound, or pharmaceutically acceptable salt thereof, according to claim 1, wherein R1 is optionally substituted 6- to 10-membered aryl, optionally substituted 3- to 6-membered cycloalkenyl, or optionally substituted 5- to 10-membered heteroaryl. 3. COMPOUND, or pharmaceutically acceptable salt thereof, according to claim 1, characterized in that the compound has the structure of Formula Ig: Formula Igem where Xe and Xf are independently N or CH; and R12 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl. 4. COMPOUND, or pharmaceutically acceptable salt thereof, according to any one of claims 1 to 3, characterized in that A is optionally substituted 6-membered arylene. 5. COMPOUND, or pharmaceutically acceptable salt thereof, according to any one of claims 1 to 3, characterized in that A is optionally substituted 5- to 6-membered heteroarylene. 6. COMPOUND, or pharmaceutically acceptable salt thereof, according to any one of claims 1 to 3, characterized in that A is optionally substituted C1-C4 heteroalkylene. 7. COMPOUND, or pharmaceutically acceptable salt thereof, according to any one of claims 1 to 3, characterized in that A is optionally substituted 3- to 6-membered heterocycloalkylene. 8. COMPOUND, or pharmaceutically acceptable salt thereof, according to any one of claims 1 to 7, characterized in that B is -CHR9-. 9. The compound, or pharmaceutically acceptable salt thereof, of claim 8, wherein R9 is F, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl. 10. COMPOUND, or pharmaceutically acceptable salt thereof, according to any one of claims 1 to 7, characterized in that B is optionally substituted 6-membered arylene. 11. The compound, or pharmaceutically acceptable salt thereof, according to claim 10, characterized in that B is 6-membered arylene. 12. COMPOUND, or a pharmaceutically acceptable salt thereof, according to any one of claims 1 to 11, characterized in that the ligand is acyclic. 13. The compound, or a pharmaceutically acceptable salt thereof, according to claim 12, characterized in that the ligand has the structure of Formula IIa: Formula IIa in which Xa is absent or is N; R14 is absent, is hydrogen or optionally substituted C1-C6 alkyl; and L2 is absent, is -SO2-, optionally substituted C1-C4 alkylene or optionally substituted C1-C4 heteroalkylene, in which at least one of Xa, R14 or L2 is present. 14. COMPOUND, or a pharmaceutically acceptable salt thereof, according to any one of claims 1 to 11, characterized in that the binder is or comprises a cyclic moiety. 15. The compound, or a pharmaceutically acceptable salt thereof, of claim 14, wherein the ligand has the structure of Formula IIb: Formula IIwhere o is 0 or 1; R15 is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted 3- to 8-membered cycloalkylene, or optionally substituted 3- to 8-membered heterocycloalkylene; X4 is absent, is optionally substituted C1-C4 alkylene, O, NCH3, or optionally substituted C1-C4 heteroalkylene; Cy is optionally substituted 3- to 8-membered cycloalkylene, optionally substituted 3- to 8-membered heterocycloalkylene, optionally substituted 6- to 10-membered arylene, or optionally substituted 5- to 10-membered heteroarylene; and L3 is absent, is -SO2-, optionally substituted C1-C4 alkylene, or optionally substituted C1-C4 heteroalkylene. 16. The compound, or a pharmaceutically acceptable salt thereof, according to any one of claims 1 to 15, characterized in that W has the structure of Formula IIIa: Formula IIIawherein R16a, R16b, and R16c are independently hydrogen, -CN, halogen, or -C1-C3 alkyl optionally substituted by one or more substituents independently selected from -OH, -O-C1-C3 alkyl, -NH2, -NH(C1-C3 alkyl), -N(C1-C3 alkyl)2, or a 4- to 7-membered saturated heterocycloalkyl. 17. The compound, or a pharmaceutically acceptable salt thereof, according to any one of claims 1 to 15, characterized in that W has the structure of Formula IIIb: Formula IIIwhere R17 is hydrogen, -C1-C3 alkyl optionally substituted by one or more substituents independently selected from -OH, -O-C1-C3 alkyl, -NH2, -NH(C1-C3 alkyl), -N(C1-C3 alkyl)2, or a 4- to 7-membered saturated cycloalkyl, or a 4- to 7-membered saturated heterocycloalkyl. 18. The compound, or a pharmaceutically acceptable salt thereof, according to any one of claims 1 to 15, characterized in that W has the structure of Formula IIIc: Formula IIIc wherein R18a, R18b, and R18c are independently hydrogen, -CN or -C1-C3 alkyl optionally substituted by one or more substituents independently selected from -OH, -O-C1-C3 alkyl, -NH2, -NH(C1-C3 alkyl), -N(C1-C3 alkyl)2, or a 4- to 7-membered saturated heterocycloalkyl. 19. The compound, or a pharmaceutically acceptable salt thereof, according to any one of claims 1 to 15, characterized in that W has the structure of Formula IIId: Formula III wherein R19 is hydrogen, -C1-C3 alkyl optionally substituted by one or more substituents independently selected from OH, -O-C1-C3 alkyl, -NH2, -NH(C1-C3 alkyl), -N(C1-C3 alkyl)2, or a 4- to 7-membered saturated heterocycloalkyl, or a 4- to 7-membered saturated heterocycloalkyl. 20. COMPOUND, or a pharmaceutically acceptable salt thereof, according to claim 1, characterized in that it is selected from: 21. PHARMACEUTICAL COMPOSITION, characterized by the fact that it comprises a compound, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 20, and a pharmaceutically acceptable excipient. 22. USE OF A COMPOUND, as defined in any one of claims 1 to 20, or of a pharmaceutical composition, as defined in claim 21, characterized in that it is for the manufacture of a medicament to treat cancer. 23. USE, according to claim 22, characterized by the fact that the cancer is pancreatic cancer, colorectal cancer, non-small cell lung cancer or endometrial cancer. 24. USE according to any one of claims 22 to 23, characterized in that the cancer comprises a Ras mutation. 25. USE, according to claim 24, characterized by the fact that the Ras mutation is K-Ras G12C, K-Ras G13C, H-Ras G12C, H-Ras G13C, N-Ras G12C or N-Ras G13C. 26. USE OF A COMPOUND, as defined in any one of claims 1 to 20, or of a pharmaceutical composition, as defined in claim 21, characterized in that it is for the manufacture of a medicament to treat a disorder related to the Ras protein.