CN121419769A

CN121419769A - Heterofunctional compounds and their usage

Info

Publication number: CN121419769A
Application number: CN202480042525.3A
Authority: CN
Inventors: G·R·克拉布特里; R·C·萨罗特; M·N·尼克斯; 张廷虎; N·S·格雷; S·古里桑卡尔; A·克罗霍京
Original assignee: Leland Stanford Junior University
Current assignee: Leland Stanford Junior University
Priority date: 2023-06-30
Filing date: 2024-06-28
Publication date: 2026-01-27
Also published as: WO2025007026A1; AU2024308508A1

Abstract

Disclosed herein are heterobifunctional compounds comprising a BCL6 binding moiety and a moiety that binds to the bromodomain of Histone Acetyltransferase (HAT), wherein the two moieties are connected by a linker. Also disclosed herein are pharmaceutical compositions comprising the compounds, and methods of using the compounds, e.g., to treat proliferative diseases such as cancer.

Description

Heterobifunctional compounds and methods of use thereof

Cross Reference to Related Applications

The present application claims priority and benefit from U.S. provisional patent application No. 63/511,374 filed on month 6 of 2023, 30, which is incorporated herein by reference in its entirety.

Statement regarding federally sponsored research

The present invention was completed under government support under contracts CA163915, CA276167 and MH126720 awarded by the national institutes of health (National Institutes of Health). The government has certain rights in this invention.

Statement of sequence Listing

The contents of the electronic sequence listing (size: 2,854 bytes; and date of creation: 2024, 6, 28 days) titled STDU2_42137_601_sequence listing.

Technical Field

Background

One of the major achievements in cancer biology in the past 40 years has been the identification of cancer drivers that drive proliferation, diffusion or survival of cancer cells upon mutation or other deregulation (Weinberg, science 230 (4727), 770-776 (1985), davoli et al Cell 155, 948-962 (2013), sanchez-Vega et al Cell 173, 321-337 e310 (2018), denny et al Cell 166, 328-342 (2016)). Cancer driving factors have been widely recognized by human genetic studies of specific tumors and can be classified as tumor suppressors that prevent the development of cancer, or oncogenes that directly promote the proliferation or spread of cancer at the expense of organisms.

Parallel studies, conducted primarily in the laboratory of developmental biologists, have identified a pathway of programmed Cell death that kills specific Cell populations that are faulty in the developmental process or faulty in DNA replication or repair (HENGARTNER et al Cell 76, 665-676 (1994); straser et al annu. Rev. Biochem. 69, 217-245 (2000)). These cell death pathways have evolved over the years and are very effective. In fact, about 600 million autologous cells per day are killed by these mechanisms (Alberts et al in Molecular Biology of the Cell (2002)). Genes, proteins and biochemical pathways that induce (pro-apoptotic) and prevent (anti-apoptotic) programmed cell death are well known and well understood.

Disclosure of Invention

Disclosed herein are compounds of formula (I):

A-L-B (I)

Or a pharmaceutically acceptable salt thereof, wherein:

a is a BCL6 binding moiety;

L is a linker, and

B is the moiety that binds to the bromodomain of histone acetyltransferase.

In some embodiments, a is a moiety of the formula:

、、

、、、

、 Or (b) 。

In some embodiments, a is a moiety of the formula:

。

in some embodiments, B is a moiety that binds to the bromodomain of p300 or CBP. In some embodiments, B is a moiety of the formula:

、、、、 Or (b) 。

In some embodiments, B is a moiety of the formula:

。

in some embodiments, the compound is a compound of formula (Ia):

(Ia)。

In some embodiments, L is a direct bond, or comprises any combination of-CH ₂-、-CH=CH-、-C≡C-、-O-、-NR'-、-BR'-、-S-、-C(O)-、-C(NR')-、-S(O)-、-S(O)₂ -, arylene, heteroarylene, cycloalkylene, and heterocyclylene moieties, wherein the arylene, heteroarylene, cycloalkylene, and heterocyclylene moieties are independently unsubstituted or substituted with 1, 2, or 3 substituents. In some embodiments, L is a direct bond, or comprises any combination of-CH ₂-、-O-、-C(O)-、-NH-、-N(CH₃) -, cycloalkylene, and heterocyclylene moieties. In some embodiments, L comprises any combination of the following moieties:

、-(CH₂)_q-、-C(O)-、-NH-、、、、、、,

wherein p is 1,2, 3, 4,5 or 6 and q is 1,2, 3, 4,5, 6,7, 8, 9, 10, 11, 12, 13, 14, 15 or 16. In some embodiments, L has a formula selected from the group consisting of:

、 And ;

Wherein p is 1,2, 3,4, 5 or 6 and q is 1,2, 3,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16.

In some embodiments, L has a formula selected from the group consisting of:

And 。

In some embodiments, the compound is selected from:

and pharmaceutically acceptable salts thereof.

Also disclosed herein is a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

Also disclosed herein is a method of treating a proliferative disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof. In some embodiments, the proliferative disease is a cancer selected from the group consisting of carcinoma, sarcoma, and hematological malignancy. In some embodiments, the cancer is a lymphoma. In some embodiments, the diffuse large B-cell lymphoma.

Also disclosed herein is a compound of formula (I) or a pharmaceutically acceptable salt thereof, for use as a medicament.

Also disclosed herein is a compound of formula (I) or a pharmaceutically acceptable salt thereof, for use in the treatment of a proliferative disease. In some embodiments, the proliferative disease is a cancer selected from the group consisting of carcinoma, sarcoma, and hematological malignancy. In some embodiments, the cancer is a lymphoma. In some embodiments, the cancer is diffuse large B-cell lymphoma.

Also disclosed herein is a kit comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof.

Drawings

FIG. 1 shows a general strategy for activating the cell death pathway with cancer drivers. The main cancer driver pathway is shown on the left. The right side shows pro-apoptotic or cell death pathways. TCIP binds the cancer driver pathway to one chemical entity and the cell death pathway to a second chemical entity. In the context of this application, histone acetyltransferases provide a driving mechanism for certain cancers, such as diffuse large B-cell lymphoma (DLBCL), and BCL6 is a potent regulator of programmed cell death or apoptosis.

FIG. 2 is a schematic representation of a transcription/epigenetic chemical proximity inducer (TCIP) that binds histone acetyltransferase on one side and BCL6 on the other side. The combination of these two proteins with TCIP is known as a ternary complex.

Fig. 3 is a schematic diagram showing TCIP borrowing only a small portion of the total target and reconnecting to create a new connection. This strategy of functional acquisition is the basis for their remarkable efficacy and lack of toxicity.

Fig. 4 shows activation of reporter Green Fluorescent Protein (GFP) under transcriptional control of endogenous BCL6 in diffuse large B-cell lymphoma (DLBCL) cell line KARPAS 422. Cells were treated with HAT-BCL6 TCIP with drug for 24 hours. Mean ± standard error, n=1-3 independent replicates.

FIGS. 5A-5B show the effect of HAT-BCL6 TCIP on viability of the DLBCL family SUDHL5 (FIG. 5A) and KARPAS422 (FIG. 5B). Cells were treated with the compound for 72 hours. Mean ± standard deviation, 1-3 biological replicates.

FIGS. 6A through 6B show the results of measuring cell viability after competitive titration of BCL6 or p300/CBP monovalent inhibitors with 1 nM of the most potent HAT-BCL6 TCIP MNN-03-038 in either the SUDHL5 (FIG. 6A) or KARPAS422 (FIG. 6B) cell lines. Cells were treated with the compound for 72 hours.

FIGS. 7A to 7G show the results of measuring oncoprotein c-MYC suppression by (FIG. 7A) measuring protein level western blot of c-MYC, BCL6, CBP and p300 after 4 hours MNN-03-038 (TCIP 3) treatment in SUDHL5 cells. Representing 3 biological replicates. (fig. 7B) quantification of western blot in fig. 7A, mean ± standard deviation, n=3 biological replicates. (FIG. 7C) Western blot of time-dependent changes in C-MYC protein after 1 nM MNN-03-038 (TCIP 3) in SUDHL5 cells, representing 3 biological replicates. (fig. 7D) quantification of western blot in fig. 7C was superimposed with RT-qPCR measurements of time-dependent changes in C-MYC mRNA after 1 nM MNN-03-038 (TCIP 3) in SUDHL5 cells, the half-life suppression was calculated by fitting a monophasic exponential decay, mean ± standard deviation, n=3 biological replicates. (FIG. 7E) all differential transcripts quantified by RNA-seq after treatment with 1 nM TCIP3 in SUDHL5 cells, 3 biological replicates, P-values calculated by double-sided Wald assay and multiple comparisons adjusted by Benjamini-Hochberg. (FIG. 7F) RT-qPCR quantification of c-MYC mRNA from 1 nM MNN-03-038 (TCIP 3) treatment versus 1 nM GNE-781 and 1 nM BI-3812 treatment was compared in SUDHL5 cells. Mean ± standard deviation, n=3 biological replicates. (FIG. 7G) compares RT-qPCR quantification of 1 nM MNN-03-038 (TCIP 3) treated with increased amounts of c-MYC mRNA of BCL6 in lymphoma cell lines. Mean ± standard deviation, n=3 biological replicates.

FIGS. 8A to 8F show cytotoxicity and ternary complex formation activity by (FIG. 8A) MNN-03-038 (TCIP 3) and inhibitor activity curves in SUDHL5 cells for 72 hours, mean.+ -. Standard deviation, n=3 biological replicates. (FIG. 8B) IC ₅₀ values of 72 hour viability assay of all synthetic TCIPs in SUDHL5 cells. (FIG. 8C) activation of BCL6 reporter gene after treatment with MNN-03-038 (TCIP 3) and inhibitor, mean.+ -. Standard deviation; n=3 biological replicates. (FIG. 8D) TR-FRET assay shows a ternary complex between recombinant p 300-bromodomain-His and BCL6-BTB domain protein after 1 hour co-incubation with TCIP molecule. (FIG. 8E) 72 hour competitive titration of p300/CBP inhibitor (GNE-781 or SGC-CBP-30) or BCL6 inhibitor (BI-3812, GSK 137 or CCT 373566) with MNN-03-038 (TCIP 3) (1 nM) in SUDHL5 cells, mean.+ -. Standard deviation; n=2-3 biological replicates. (FIG. 8F) 72 hour viability curves of MNN-03-038 (TCIP 3) or GNE-781+BI-3812 treatment in four DLBCL cell lines to reduce BCL6 levels.

Detailed Description

The fact that the biochemical pathways used to drive cancer coexist in the same cell as the programmed cell death pathways of the active killer cells opens the possibility that one can reconnect the cellular biochemical circuit so that cancer cells kill themselves using the biochemical pathways initiated by the cancer driver. Disclosed herein is a class of small molecules (transcriptional/epigenetic chemical proximity inducers, or TCIP) that reconnect the biochemical circuit of cells to activate pathways that are normally not controlled by the first pathway, in this case cancer drivers. The operation of these molecules is shown by grey arrows in fig. 1, which connect the cancer driver and its dependent biochemical pathways with the cell death pathways, in this case initiated by DNA replication damage or developmental errors. FIG. 2 shows the general structure of TCIP, which recruits the transcriptional activator histone acetyltransferase to the promoter of the BCL 6-bound cell death gene.

TCIP borrows only a portion of the target protein and uses it to activate transcription. For example, as shown in FIG. 3, only a portion of the total HAT is used. By borrowing only a small amount of target protein, rather than inhibiting all of it, TCIP can avoid the mechanism-based toxicity associated with completely inhibiting or eliminating the target. This is generally shown in fig. 3.

Disclosed herein are high-efficiency TCIP for treating cancer, such as human diffuse large B-cell lymphoma (DLCBL). BCL6 suppresses pro-apoptotic genes by binding to histone deacetylase. Thus, recruitment of histone acetyltransferases may activate these suppressed genes. A compound such as B-I3812 binds to the BTB domain of BCL6 and prevents histone deacetylase binding. Thus, the compounds disclosed herein are designed to bind to the BTB domain of BCL6 and are chemically linked to entities that bind Histone Acetyltransferase (HAT) Ep300 and CBP.

Definition of the definition

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In case of conflict, the present document, including definitions, will control.

The definition of specific functional groups and chemical terms is described in more detail below. For purposes of this disclosure, chemical elements are identified according to the CAS version, handbook of CHEMISTRY AND PHYSICS, 75 th edition, inner cover, and specific functional groups are generally defined as described herein. In addition, the general principles of organic chemistry and specific functional moieties and reactivities are described in Sorrell, organic Chemistry, 2 nd , University Science Books, Sausalito, 2006;Smith, March's Advanced Organic Chemistry: Reactions, Mechanism, and Structure, th edition, 7 th edition, john Wiley & Sons, inc., new York, 2013; larock, comprehensive Organic Transformations, 3 rd edition, john Wiley & Sons, inc., new York, 2018, and Carruthers, some Modern Methods of Organic Synthesis, 3 rd edition, cambridge University Press, cambridge, 1987.

As used herein, the modifier "about" used in connection with a quantity is inclusive of the stated value and has the meaning dictated by the context (e.g., it includes at least the degree of error associated with measurement of the particular quantity). The modifier "about" should also be considered as disclosing the range defined by the absolute values of the two endpoints. For example, the expression "about 2 to about 4" also discloses the range "2 to 4". The term "about" may refer to + -10% of the specified number. For example, "about 10%" may indicate a range of 9% to 11%, and "about 1" may mean 0.9-1.1. Other meanings of "about" may be evident from the context, such as rounding, for example, "about 1" may also mean 0.5 to 1.4.

As used herein, the term "alkyl" refers to a straight or branched saturated hydrocarbon chain group. The alkyl chain may contain, for example, 1 to 24 carbon atoms (C ₁-C₂₄ alkyl), 1 to 16 carbon atoms (C ₁-C₁₆ alkyl), 1 to 14 carbon atoms (C ₁-C₁₄ alkyl), 1 to 12 carbon atoms (C ₁-C₁₂ alkyl), 1 to 10 carbon atoms (C ₁-C₁₀ alkyl), 1 to 8 carbon atoms (C ₁-C₈ alkyl), 1 to 6 carbon atoms (C ₁-C₆ alkyl), 1 to 4 carbon atoms (C ₁-C₄ alkyl), 1 to 3 carbon atoms (C ₁-C₃ alkyl), or 1 to 2 carbon atoms (C ₁-C₂ alkyl). Representative examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2-dimethylpentyl, 2, 3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl and dodecyl.

As used herein, the term "alkenyl" refers to a straight or branched hydrocarbon chain group containing at least one carbon-carbon double bond and no triple bonds. The one or more double bonds may be located at any one or more positions of the hydrocarbon chain. Alkenyl chains may contain, for example, 2 to 24 carbon atoms (C ₂-C₂₄ alkenyl), 2 to 16 carbon atoms (C ₂-C₁₆ alkenyl), 2 to 14 carbon atoms (C ₂-C₁₄ alkenyl), 2 to 12 carbon atoms (C ₂-C₁₂ alkenyl), 2 to 10 carbon atoms (C ₂-C₁₀ alkenyl), 2 to 8 carbon atoms (C ₂-C₈ alkenyl), 2 to 6 carbon atoms (C ₂-C₆ alkenyl), 2 to 4 carbon atoms (C ₂-C₄ alkenyl), 2 to 3 carbon atoms (C ₂-C₃ alkenyl) or 2 carbon atoms (C ₂ alkenyl). Representative examples of alkenyl groups include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, butadienyl, 2-methyl-2-propenyl, 3-butenyl, pentenyl, pentadienyl, hexenyl, heptenyl, octenyl, xin San alkenyl, and the like.

As used herein, the term "alkynyl" means a straight or branched hydrocarbon chain group containing at least one carbon-carbon triple bond. Alkynyl chains may contain, for example, 2 to 24 carbon atoms (C ₂-C₂₄ alkynyl), 2 to 16 carbon atoms (C ₂-C₁₆ alkynyl), 2 to 14 carbon atoms (C ₂-C₁₄ alkynyl), 2 to 12 carbon atoms (C ₂-C₁₂ alkynyl), 2 to 10 carbon atoms (C ₂-C₁₀ alkynyl), 2 to 8 carbon atoms (C ₂-C₈ alkynyl), 2 to 6 carbon atoms (C ₂-C₆ alkynyl), 2 to 4 carbon atoms (C ₂-C₄ alkynyl), 2 to 3 carbon atoms (C ₂-C₃ alkynyl) or 2 carbon atoms (C ₂ alkynyl). The one or more triple bonds may be located at any one or more positions of the hydrocarbon chain. Representative examples of alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, and the like.

As used herein, the term "alkoxy" refers to an alkyl group, as defined herein, appended to the parent molecular moiety through an oxygen atom. Representative examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, 2-propoxy, butoxy, and t-butoxy.

As used herein, the term "amino" refers to the group-NR _xR_y, wherein R _x and R _y are selected from hydrogen and alkyl (e.g., C ₁-C₄ alkyl). The group-NH (alkyl) may be referred to herein as "alkylamino", and the group-N (alkyl) ₂ may be referred to herein as "dialkylamino".

As used herein, the term "aryl" refers to a monocyclic, bicyclic, or tricyclic 4n+2 aromatic ring system (e.g., having 6, 10, or 14 pi electrons shared in a cyclic array) having 6-14 ring carbon atoms and zero heteroatoms ("_C6-_C14 aryl"). In some embodiments, aryl groups have six ring carbon atoms ("_C6 aryl", i.e., phenyl). In some embodiments, aryl groups have ten ring carbon atoms ("_C10 aryl", for example, naphthyl such as 1-naphthyl and 2-naphthyl). In some embodiments, aryl groups have fourteen ring carbon atoms ("_C14 aryl", i.e., anthracenyl and phenanthrenyl).

As used herein, the term "arylene" refers to a divalent aryl group.

As used herein, the term "cycloalkyl" refers to a saturated carbocyclic ring system group containing three to ten carbon atoms and zero heteroatoms. Cycloalkyl groups may be monocyclic, bicyclic, bridged, fused or spiro. Representative examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, adamantyl, bicyclo [2.2.1] heptyl, bicyclo [3.2.1] octyl, and bicyclo [5.2.0] nonyl.

As used herein, the term "cycloalkylene" refers to a divalent cycloalkyl group.

As used herein, the term "cyano" refers to a —cn group.

As used herein, the term "halogen" or "halo" refers to F, cl, br or I.

As used herein, the term "haloalkyl" refers to an alkyl group as defined herein wherein at least one hydrogen atom (e.g., one, two, three, four, five, six, seven, or eight hydrogen atoms) is replaced with a halogen. In some embodiments, each hydrogen atom of the alkyl group is replaced with a halogen ("perhaloalkyl"). Representative examples of haloalkyl include, but are not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, 2-fluoroethyl, 2-trifluoroethyl and 3, 3-trifluoropropyl.

As used herein, the term "haloalkoxy" refers to a haloalkyl group as defined herein appended to the parent molecular moiety through an oxygen atom. Representative examples of haloalkoxy groups include, but are not limited to, difluoromethoxy, trifluoromethoxy, and 2, 2-trifluoroethoxy.

As used herein, the term "heteroalkyl" refers to an alkyl group, as defined herein, wherein one or more carbon atoms (and any associated hydrogen atoms) are each independently replaced by a heteroatom group such as-NH-, -O-, -S (O) ₂-、-OP(O)(O^-) O-, and the like. For example, 1, 2, 3, 4, 5, 6 or more carbon atoms may independently be replaced by the same or different heteroatom groups. Heteroalkyl groups may also contain one or more carbonyl moieties (i.e., where the carbon atom of the alkyl group is oxidized to a-C (O) -group).

As used herein, the term "heteroaryl" refers to a 5-10 membered monocyclic or bicyclic 4n+2 aromatic ring system (e.g., having 6 or 10 pi electrons shared in a cyclic array) group having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5-10 membered heteroaryl"). In heteroaryl groups containing one or more nitrogen atoms, the point of attachment may be a carbon atom or a nitrogen atom, as the valence allows. Heteroaryl bicyclic ring systems may contain one or more heteroatoms in one or both rings. "heteroaryl" also includes ring systems in which a heteroaryl ring as defined above is fused with one or more aryl groups, wherein the point of attachment is on the aryl or heteroaryl ring, and in such cases the number of ring members indicates the number of ring members in the fused (aryl/heteroaryl) ring system. Bicyclic heteroaryl groups wherein one ring does not contain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, etc.), the point of attachment can be on either ring, i.e., a ring bearing a heteroatom (e.g., 2-indolyl) or a ring not containing a heteroatom (e.g., 5-indolyl). Exemplary 5-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyrrolyl, furanyl, and thienyl. Exemplary 5-membered heteroaryl groups containing two heteroatoms include, but are not limited to, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryl groups containing three heteroatoms include, but are not limited to, triazolyl, oxadiazolyl, and thiadiazolyl. Exemplary 5-membered heteroaryl groups containing four heteroatoms include, but are not limited to, tetrazolyl. Exemplary 6-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyridinyl. Exemplary 6-membered heteroaryl groups containing two heteroatoms include, but are not limited to, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryl groups containing three or four heteroatoms include, but are not limited to, triazinyl and tetrazinyl, respectively. Exemplary 7-membered heteroaryl groups containing one heteroatom include, but are not limited to, azapyridyl, oxapyridyl, and thiapyridyl. Exemplary 5, 6-bicyclic heteroaryl groups include, but are not limited to, indolyl, isoindolyl, indazolyl, benzotriazole, benzothienyl, isobenzothienyl, benzofuranyl, benzisotofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, benzothiadiazolyl, indolizinyl, and purinyl. Exemplary 6, 6-bicyclic heteroaryl groups include, but are not limited to, naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.

As used herein, the term "heteroarylene" refers to a divalent heteroaryl group.

As used herein, the term "heterocyclyl" refers to a 3-to 10-membered non-aromatic ring system group having a ring carbon atom and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus and silicon ("3-10 membered heterocyclyl"). In a heterocyclic group containing one or more nitrogen atoms, the point of attachment may be a carbon atom or a nitrogen atom, as long as the valence allows. The heterocyclyl may be a monocyclic ("monocyclic heterocyclyl") or a fused, bridged or spiro ring system, such as a bicyclic system ("bicyclic heterocyclyl"), and may be saturated or may be partially unsaturated. The heterocyclyl bicyclic ring system may contain one or more heteroatoms in one or both rings. "heterocyclyl" also includes ring systems in which a heterocyclyl ring as defined above is fused to one or more cycloalkyl groups, in which the point of attachment is on the cycloalkyl or heterocyclyl ring, or ring systems in which a heterocyclyl ring as defined above is fused to one or more aryl or heteroaryl groups, in which the point of attachment is on the heterocyclyl ring, and in which case the number of ring members continues to indicate the number of ring members in the heterocyclyl ring system. Heterocyclyl groups may be described as, for example, 3-7 membered heterocyclyl groups, wherein the term "membered" refers to a non-hydrogen ring atom within the moiety, i.e., carbon, nitrogen, oxygen, sulfur, boron, phosphorus, and silicon. Exemplary 3-membered heterocyclic groups containing one heteroatom include, but are not limited to, aziridinyl, oxetanyl and thiiranyl (thiorenyl). Exemplary 4-membered heterocyclic groups containing one heteroatom include, but are not limited to, azetidinyl, oxetanyl, and thietanyl. Exemplary 5-membered heterocyclic groups containing one heteroatom include, but are not limited to, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2, 5-dione. Exemplary 5-membered heterocyclic groups containing two heteroatoms include, but are not limited to, dioxolanyl, oxathiolanyl (oxasulfuranyl), dithiolane (disulfuranyl), and oxazolidin-2-one. Exemplary 5-membered heterocyclic groups containing three heteroatoms include, but are not limited to, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclic groups containing one heteroatom include, but are not limited to, piperidinyl (e.g., 2, 6-tetramethylpiperidinyl), tetrahydropyranyl, dihydropyridinyl, pyridonyl (e.g., 1-methylpyridin-2-onyl), and thialkyl. Exemplary 6-membered heterocyclic groups containing two heteroatoms include, but are not limited to, piperazinyl, morpholinyl, pyridazinonyl (2-methylpyridazin-3-onyl), pyrimidinonyl (e.g., 1-methylpyrimidin-2-onyl, 3-methylpyrimidin-4-onyl), dithianyl, dioxanyl. Exemplary 6-membered heterocyclic groups containing two heteroatoms include, but are not limited to, triazinylalkyl groups. Exemplary 7-membered heterocyclyl groups containing one heteroatom include, but are not limited to, azepanyl, oxepinyl, and thiepanyl. Exemplary 8-membered heterocyclic groups containing one heteroatom include, but are not limited to, azacyclooctyl, oxacyclooctyl, and thiacyclooctyl. Exemplary 5-membered heterocyclyl groups (also referred to herein as 5, 6-bicyclic heterocyclyl rings) fused to a C ₆ aromatic ring include, but are not limited to, indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, benzoxazolinonyl, and the like. Exemplary 5-membered heterocyclyl groups fused to a heterocyclyl ring (also referred to herein as a5, 5-bicyclic heterocyclyl ring) include, but are not limited to, octahydropyrrolo-pyrrolyl (e.g., octahydropyrrolo [3,4-c ] pyrrolyl), and the like. Exemplary 6-membered heterocyclyl groups fused to a heterocyclyl ring (also referred to as a 4, 6-membered heterocyclyl ring) include, but are not limited to, diazaspiro nonyl (e.g., 2, 7-diazaspiro [3.5] nonyl). Exemplary 6-membered heterocyclyl groups fused to an aromatic ring (also referred to herein as 6, 6-bicyclic heterocyclyl rings) include, but are not limited to, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and the like. Exemplary 6-membered heterocyclyl groups fused to a cycloalkyl ring (also referred to herein as a 6, 7-bicyclic heterocyclyl ring) include, but are not limited to, azabicyclooctyl (e.g., (1, 5) -8-azabicyclo [3.2.1] octyl). Exemplary 6-membered heterocyclyl groups fused to a cycloalkyl ring (also referred to herein as a 6, 8-bicyclic heterocyclyl ring) include, but are not limited to, azabicyclo nonyl (e.g., 9-azabicyclo [3.3.1] nonyl).

As used herein, the term "heterocyclylene" refers to a divalent heterocyclic group.

As used herein, the term "hydroxyl" refers to an-OH group.

As used herein, the term "nitro" refers to a-NO ₂ group.

When a group or moiety may be substituted, the term "substituted" indicates that one or more (e.g., 1, 2, 3, 4, 5, or 6; in some embodiments 1, 2, or 3; and in other embodiments 1 or 2) hydrogen atoms on the group indicated in the expression using "substituted" may be replaced by a series of the indicated groups or by suitable substituents known to those skilled in the art (e.g., one or more of the groups listed below), provided that the normal valency of the indicated atom is not exceeded. Substituents include, but are not limited to, alkyl, alkenyl, alkynyl, alkoxy, acyl, amino, amido, amidino, aryl, azido, carbamoyl, carboxyl ester, cyano, cycloalkyl, cycloalkenyl, guanidino, halo, haloalkyl, haloalkoxy, heteroalkyl, heteroaryl, heterocyclyl, hydroxy, hydrazino, imino, oxo, nitro, phosphate, phosphonate, sulfonic acid, thiol, thioketone, or combinations thereof.

As used herein, in chemical structure, the indication:

representing the point of attachment of one moiety to another moiety (e.g., substituent to the remainder of the compound).

For the compounds described herein, the groups and substituents thereof may be selected according to the permissible valences of atoms and substituents such that the selections and substitutions result in stable compounds, e.g., which do not spontaneously undergo transformations such as by rearrangement, cyclization, elimination, and the like.

Such indications also encompass substituents resulting from right-to-left writing structures when the substituents are specified by their conventional formulas written from left-to-right. For example, if a divalent group is shown as-CH ₂ O-, such an indication also encompasses-OCH ₂ -; similarly, -OC (O) NH-also encompasses-NHC (O) O-. When the linker moiety is shown, the linker may be attached to the other moiety of the compound in either direction.

The terms "administration (administer)", "Administration (ADMINISTERING)", or "administration" as used herein refer to implantation, absorption, ingestion, injection, inhalation, or otherwise introducing a compound or pharmaceutical composition.

As used herein, the terms "disorder," "disease," and "condition" are used interchangeably.

An "effective amount" of a compound or composition refers to an amount sufficient to elicit the desired biological response (e.g., to treat a disorder). Those of skill in the art will appreciate that the effective amount of a compound may vary depending on factors such as the desired biological endpoint, the pharmacokinetics of the compound, the condition being treated, the mode of administration, and the age and health of the subject. An effective amount encompasses both therapeutic and prophylactic treatment. For example, in treating cancer, an effective amount of a compound or composition may reduce the burden of a tumor or prevent the growth or spread of a tumor.

A "therapeutically effective amount" of a compound or composition refers to an amount sufficient to provide a therapeutic benefit in treating a disorder or delay or minimize one or more symptoms associated with the disorder. In some embodiments, a therapeutically effective amount is an amount sufficient to provide a therapeutic benefit in treating a disorder or to minimize one or more symptoms associated with the disorder. A therapeutically effective amount of a compound means an amount of a therapeutic agent that provides a therapeutic benefit in treating a disorder, either alone or in combination with other therapies. The term "therapeutically effective amount" may encompass an amount that improves overall therapy, alleviates symptoms of the disorder or avoids the cause of the disorder, or enhances the therapeutic efficacy of another therapeutic agent.

"Subject" contemplated for administration includes, but is not limited to, humans (i.e., males or females of any age group, e.g., pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young, middle-aged, or elderly) and/or other non-human animals, e.g., mammals (e.g., primates (e.g., cynomolgus, rhesus); commercially relevant mammals such as cows, pigs, horses, sheep, goats, cats, and/or dogs) and birds (e.g., commercially relevant birds such as chickens, ducks, geese, and/or turkeys).

As used herein, the terms "treating", "treatment" and "treatment" refer to reversing, alleviating, delaying the onset of, or inhibiting the progression of a disease or disorder or one or more signs or symptoms thereof. In some embodiments, "treatment" and "treatment" require that signs or symptoms of the disease, disorder or condition have been developed or observed. In other embodiments, the treatment may be administered without signs or symptoms of the disease or disorder. For example, treatment may be administered to a susceptible individual prior to onset of symptoms (e.g., based on a history of symptoms and/or based on genetic or other susceptibility factors). Treatment may also continue after symptom relief, for example, to delay or prevent recurrence.

Compounds of formula (I)

Disclosed herein are compounds of formula (I):

A-L-B (I)

Or a pharmaceutically acceptable salt thereof, wherein:

a is a BCL6 binding moiety;

L is a linker, and

B is the moiety that binds to the bromodomain of histone acetyltransferase.

Part a is a BCL6 binding moiety. In some embodiments, a is a moiety of the formula:

、、

、、、

、 Or (b) 。

In some embodiments, a is a moiety of the formula:

。

Part B is the part that binds to the bromodomain of histone acetyltransferase. In some embodiments, B is a moiety that binds to the bromodomain of p300 or CBP. In some embodiments, B is a moiety of the formula:

、、、

、 Or (b) 。

In some embodiments, B is a moiety of the formula:

。

in some embodiments, the compound of formula (I) is a compound of formula (Ia):

(Ia),

or a pharmaceutically acceptable salt thereof.

L is a linker that provides a covalent linkage between the a and B moieties. In some embodiments, the exact structure of the linker may not be critical, so long as it does not substantially interfere with the activity of the BCL6 binding moiety or the moiety that binds to the bromodomain of HAT. In some embodiments, L is a direct bond. In other embodiments, L comprises any combination of-CH ₂-、-CH=CH-、-C≡C-、-O-、-NR'-、-BR'-、-S-、-C(O)-、-C(NR')-、-S(O)-、-S(O)₂ -, arylene, heteroarylene, cycloalkylene, and heterocyclylene moieties, wherein R' is selected from hydrogen and C ₁-C₆ alkyl, and wherein the arylene, heteroarylene, cycloalkylene, and heterocyclylene moieties are independently unsubstituted or substituted with 1, 2, or 3 substituents.

In some embodiments, the linker comprises an alkylene chain (e.g., having 2-20-CH ₂ -units). In other embodiments, the linker comprises an alkylene chain interrupted and/or terminated by at least one group selected from (at either or both termini )：-O-、-S-、-N(R')-、-CH=CH-、-C≡C-、-C(O)-、-C(O)O-、-OC(O)-、-OC(O)O-、-C(NOR')-、-C(O)N(R')-、-C(O)N(R')C(O)-、-C(O)N(R')C(O)N(R')-、-N(R')C(O)-、-N(R')C(O)N(R')-、-N(R')C(O)O-、-OC(O)N(R')-、-C(NR')-、-N(R')C(NR')-、-C(NR')N(R')-、-N(R')C(NR')N(R')-、-OB(CH₃)O-、-S(O)₂-、-OS(O)-、-S(O)O-、-S(O)-、-OS(O)₂-、-S(O)₂O-、-N(R')S(O)₂-、-S(O)₂N(R')-、-N(R')S(O)-、-S(O)N(R')-、-N(R')S(O)₂N(R')-、-N(R')S(O)N(R')-、C₃-C₁₂ cycloalkylene, 3-to 12-membered heterocyclylene, 5-to 10-membered arylene, 5-to 12-membered heteroarylene, or any combination thereof, wherein each R' is independently selected from hydrogen and C ₁-C₆ alkyl, and wherein the interrupting and terminating groups may be the same or different.

In some embodiments, L comprises an alkylene chain: Wherein q is 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16. In some embodiments, q is 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12. For example, in some embodiments, q is 1-16、1-15、1-14、1-13、1-12、1-11、1-10、1-9、1-8、1-7、1-6、1-5、1-4、1-3、1-2、2-16、2-15、2-14、2-13、2-12、2-11、2-10、2-9、2-8、2-7、2-6、2-5、2-4、2-3、3-16、3-15、3-14、3-13、3-12、3-11、3-10、3-9、3-8、3-7、3-6、3-5、3-4、4-16、4-15、4-14、4-13、4-12、4-11、4-10、4-9、4-8、4-7、4-6、4-5、5-16、5-15、5-14、5-13、5-12、5-11、5-10、5-9、5-8、5-7、5-6、6-16、6-15、6-14、6-13、6-12、6-11、6-10、6-9、6-8、6-7、7-16、7-15、7-14、7-13、7-12、7-11、7-10、7-9、7-8、8-16、8-15、8-14、8-13、8-12、8-11、8-10、8-9 or 9-16、9-15、9-14、9-13、9-12、9-11、9-10、10-16、10-15、10-14、10-13、10-12、10-11、11-16、11-15、11-14、11-13、11-12、12-16、12-15、12-14、12-13、13-16、13-15、13-14、14-16、14-15 and 15-16. In some embodiments, q is 1. In some embodiments, q is 2. In some embodiments, q is 3. In some embodiments, q is 4. In some embodiments, q is 5. In some embodiments, q is 6. In some embodiments, q is 7. In some embodiments, q is 8. In some embodiments, q is 9. In some embodiments, q is 10. In some embodiments, q is 11. In some embodiments, q is 12. In some embodiments, q is 13. In some embodiments, q is 14. In some embodiments, q is 15. In some embodiments, q is 16. Specific examples of alkylene chains include:

、、、、、、

、、、

、、

、 And

。

In some embodiments, L comprises an alkylene chain interrupted by a functional group, such as -C(O)-、-C(O)O-、-OC(O)-、-OC(O)O-、-C(NOR')-、-C(O)N(R')-、-C(O)N(R')C(O)-、-C(O)N(R')C(O)N(R')-、-N(R')C(O)-、-N(R')C(O)N(R')-、-N(R')C(O)O-、-OC(O)N(R')-、-C(NR')-、-N(R')C(NR')-、-C(NR')N(R')-、-N(R')C(NR')N(R')-、-OB(CH₃)O-、-S(O)₂-、-OS(O)-、-S(O)O-、-S(O)-、-OS(O)₂-、-S(O)₂O-、-N(R')S(O)₂-、-S(O)₂N(R')-、-N(R')S(O)-、-S(O)N(R')-、-N(R')S(O)₂N(R')- or-N (R ') S (O) N (R') -).

In some embodiments, L comprises one or more alkylene glycol repeat units, such as ethylene glycol or propylene glycol repeat units. For example, in some embodiments, L comprises a polyethylene glycol or oligoethylene glycol chain: wherein p is 1,2,3, 4, 5 or 6. In some embodiments, L comprises a set of formulas: Wherein p is 1, 2, 3, 4, 5 or 6. In some embodiments, p is 1-6, 1-5, 1-4, 1-3, 1-2, 2-6, 2-5, 2-4, 2-3, 3-6, 3-5, 3-4, 4-6, or 5-6. In some embodiments, p is 1. In some embodiments, p is 2. In some embodiments, p is 3. In some embodiments, p is 4. In some embodiments, p is 5. In some embodiments, p is 6.

In some embodiments, L comprises one or more cycloalkylene or heterocyclylene groups, non-limiting examples of which include:

、、、、、、

、、、、、

、、、、

、、 And 。

In some embodiments, L comprises a cycloalkylene or heterocyclylene group, such as one of the cycloalkyl and heterocyclylene groups described above, having one or more additional groups on one or both ends, wherein the additional groups are independently selected from -CH₂-、-O-、-S-、-N(R')-、-CH=CH-、-C≡C-、-C(O)-、-C(O)O-、-OC(O)-、-OC(O)O-、-C(NOR')-、-C(O)N(R')-、-C(O)N(R')C(O)-、-C(O)N(R')C(O)N(R')-、-N(R')C(O)-、-N(R')C(O)N(R')-、-N(R')C(O)O-、-OC(O)N(R')-、-C(NR')-、-N(R')C(NR')-、-C(NR')N(R')-、-N(R')C(NR')N(R')-、-OB(CH₃)O-、-S(O)₂-、-OS(O)-、-S(O)O-、-S(O)-、-OS(O)₂-、-S(O)₂O-、-N(R')S(O)₂-、-S(O)₂N(R')-、-N(R')S(O)-、-S(O)N(R')-、-N(R')S(O)₂N(R')- and-N (R ') S (O) N (R ') -, or any combination thereof, wherein each R ' is independently selected from hydrogen and C ₁-C₆ alkyl. In some embodiments, L comprises a cycloalkylene or heterocyclylene group, such as one of the cycloalkylene and heterocyclylene groups described above, having one or more additional groups at one or both ends, wherein the additional groups are independently selected from-CH ₂ -, -C (O) -, -NH-and-O-.

In some embodiments, L comprises two or more cycloalkylene or heterocyclylene groups, such as two or three cycloalkylene or heterocyclylene groups independently selected from those described above, that are attached to each other by a covalent bond or one or more linking groups, optionally with one or more additional linking groups on one or both ends, wherein the linking groups are independently selected from -O-、-S-、-N(R')-、-CH₂-、-CH=CH-、-C≡C-、-C(O)-、-C(O)O-、-OC(O)-、-OC(O)O-、-C(NOR')-、-C(O)N(R')-、-C(O)N(R')C(O)-、-C(O)N(R')C(O)N(R')-、-N(R')C(O)-、-N(R')C(O)N(R')-、-N(R')C(O)O-、-OC(O)N(R')-、-C(NR')-、-N(R')C(NR')-、-C(NR')N(R')-、-N(R')C(NR')N(R')-、-OB(CH₃)O-、-S(O)₂-、-OS(O)-、-S(O)O-、-S(O)-、-OS(O)₂-、-S(O)₂O-、-N(R')S(O)₂-、-S(O)₂N(R')-、-N(R')S(O)-、-S(O)N(R')-、-N(R')S(O)₂N(R')- and-N (R ') S (O) N (R ') -, or any combination thereof, wherein each R ' is independently selected from hydrogen and C ₁-C₆ alkyl. In some embodiments, L comprises two or more cycloalkylene or heterocyclylene groups, such as two or three cycloalkylene or heterocyclylene groups independently selected from those described above, that are linked to each other by a covalent bond or one or more linking groups, optionally with one or more additional linking groups on one or both ends, wherein the linking groups are independently selected from-O-and-CH ₂ -.

In some embodiments, L comprises one or more cycloalkylene or heterocyclylene groups substituted with 1,2, or 3 substituents, e.g., substituents independently selected from C ₁-C₄ alkyl, C ₁-C₄ haloalkyl, C ₁-C₄ alkoxy, halo, hydroxy, and the like. In some embodiments, L comprises one or more cycloalkylene or heterocyclylene groups substituted with 1,2, or 3 substituents independently selected from C ₁-C₄ alkyl and halo. In some embodiments, L comprises one or more cycloalkylene or heterocyclylene groups substituted with 1,2, or 3 substituents independently selected from methyl and fluoro.

In some embodiments, L is a direct bond, or comprises any combination of-CH ₂-、-CH=CH-、-C≡C-、-O-、-NR'-、-BR'-、-S-、-C(O)-、-C(NR')-、-S(O)-、-S(O)₂ -, arylene, heteroarylene, cycloalkylene, and heterocyclylene moieties, wherein the arylene, heteroarylene, cycloalkylene, and heterocyclylene moieties are independently unsubstituted or substituted with 1,2, or 3 substituents. In some embodiments, L is a direct bond, or comprises any combination of-CH ₂-、-O-、-C(O)-、-NH-、-N(CH₃) -, cycloalkylene, and heterocyclylene moieties.

In some embodiments, L comprises any combination of the following moieties:

、-(CH₂)_q-、-C(O)-、-NH-、、、、

、 And ,

In some embodiments, L has a formula selected from the group consisting of:

、 And ;

In some embodiments, L has a formula selected from the group consisting of:

And 。

In some embodiments, the compound of formula (I) is selected from:

and pharmaceutically acceptable salts thereof.

When discussing certain features or characteristics of compounds of formula (I) herein or of compositions, methods, or kits comprising compounds of formula (I), it is to be understood that such references also include compounds of formula (Ia), as well as specific exemplary compounds disclosed herein.

The compounds may exist as stereoisomers wherein asymmetric or chiral centers are present. Stereoisomers are "R" or "S", depending on the configuration of substituents around the chiral carbon atom. The terms "R" and "S" as used herein are configurations as defined in IIUPAC 1974 Recommendations for Section E, fundamental Stereochemistry, pure appl. Chem. 1976, 45:13-30. The various stereoisomers and mixtures thereof are specifically included within the scope of the present disclosure. Stereoisomers include enantiomers and diastereomers, as well as mixtures of enantiomers or diastereomers. Individual stereoisomers of the compounds may be prepared synthetically from commercially available starting materials containing asymmetric or chiral centers or by preparing racemic mixtures followed by resolution procedures well known to those of ordinary skill in the art. Examples of such resolution methods are (1) attaching a mixture of enantiomers to a chiral auxiliary, separating the resulting mixture of diastereomers by recrystallization or chromatography, and optionally liberating optically pure products from the auxiliary, as described in Furniss, hannaford, smith, and Tatchell, "Vogel's Textbook of Practical Organic Chemistry," 5 th edition (1989), longman Scientific & Technical, essex CM202JE, england, (2) separating the mixture of optical enantiomers directly on a chiral chromatographic column, or (3) fractional recrystallization methods.

It is to be understood that the compounds may exist in different tautomeric forms and that all such forms are included within the scope of the disclosure.

The present disclosure also includes isotopically-labeled compounds, which are identical to those recited in formula (I), but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes suitable for inclusion in the compounds of the invention are hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, such as, but not limited to ²H、³H、¹³C、¹⁴C、¹⁵N、¹⁸O、¹⁷O、³¹P、³²P、³⁵S、¹⁸F and ³⁶ Cl, respectively. Substitution with heavier isotopes such as deuterium (² H) may afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements and therefore may be preferred in some circumstances. The compounds may incorporate positron emitting isotopes for use in medical imaging and Positron Emission Tomography (PET) studies to determine receptor distribution. Suitable positron emitting isotopes that can be incorporated into compounds of formula (I) are ¹¹C、¹³N、¹⁵ O and ¹⁸ F. Isotopically-labeled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art, or by processes analogous to those described in the accompanying examples, using an appropriate isotopically-labeled reagent in place of a non-isotopically-labeled reagent.

The compounds disclosed herein may exist in solvated as well as unsolvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like, and the present disclosure is intended to cover solvated and unsolvated forms. In one embodiment, the compound is amorphous. In one embodiment, the compound is a single polymorph. In another embodiment, the compound is a mixture of polymorphs. In another embodiment, the compound is in crystalline form.

A. pharmaceutically acceptable salts

The disclosed compounds may exist as pharmaceutically acceptable salts. The term "pharmaceutically acceptable salt" refers to salts or zwitterions of a compound that are water-soluble or oil-soluble or dispersible, are suitable for use in treating a condition without undue toxicity, irritation, and allergic response commensurate with a reasonable benefit/risk ratio, and are effective for their intended use. Salts may be prepared during the final isolation and purification of the compounds or separately by reacting the amino groups of the compounds with a suitable acid. For example, the compound may be dissolved in a suitable solvent such as, but not limited to, methanol and water and treated with at least one equivalent of an acid (e.g., hydrochloric acid). The resulting salt may precipitate, be isolated by filtration and dried under reduced pressure. Alternatively, the solvent and excess acid may be removed under reduced pressure to provide a salt. Representative salts include acetates, adipates, alginates, citrates, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphoric acid, camphorsulfonate, digluconate, glycerophosphate, hemisulfate, heptanoate, caproate, formate, isethionate, fumarate, lactate, maleate, methanesulfonate, naphthalenesulfonate, nicotinate, oxalate, pamoate, pectate, persulfate, 3-phenylpropionate, picrate, oxalate, maleate, pivalate (pivalate), propionate, succinate, tartrate, trichloroacetate, trifluoroacetate, glutamate, p-toluenesulfonate, undecanoate, hydrochloride, hydrobromide, sulfate, phosphate, and the like. The amino groups of the compounds may also be quaternized with alkyl chlorides, bromides and iodides such as methyl, ethyl, propyl, isopropyl, butyl, lauryl, myristyl, stearyl, and the like.

Basic addition salts can be prepared by reacting the carboxyl groups with suitable bases such as hydroxides, carbonates or bicarbonates of metal cations (such as lithium, sodium, potassium, calcium, magnesium or aluminum), or organic primary, secondary or tertiary amines, during the final isolation and purification of the disclosed compounds. Quaternary amine salts such as those derived from methylamine, dimethylamine, trimethylamine, triethylamine, diethylamine, ethylamine, tributylamine, pyridine, N-dimethylaniline, N-methylpiperidine, N-methylmorpholine, dicyclohexylamine, procaine, dibenzylamine, N-dibenzylphenylamine, 1-dibenzylhydroxylamine, and N, N' -dibenzylethylenediamine, ethylenediamine, ethanolamine, diethanolamine, piperidine, piperazine, and the like can be prepared.

B. Synthesis method

In another aspect, disclosed herein are methods for making a compound of formula (I), or a pharmaceutically acceptable salt thereof. In general, the compounds of formula (I) and pharmaceutically acceptable salts thereof may be prepared by any known method suitable for preparing chemically related compounds. Exemplary suitable synthetic schemes are provided in the examples section.

The compounds and intermediates may be isolated and purified by methods well known to those skilled in the art of organic synthesis. Examples of conventional methods for separating and purifying compounds may include, but are not limited to, chromatography on solid supports such as silica gel, alumina or silica derivatized with alkylsilane groups, by recrystallization at high or low temperatures and optional pretreatment with activated carbon, thin layer chromatography, distillation under various pressures, sublimation under vacuum, and wet milling, as described, for example, "Vogel's Textbook of Practical Organic Chemistry," 5 th edition (1989), furniss, hannaford, smith and Tatchell, pub, longman Scientific & Technical, essex CM20 JE, england.

The reaction conditions and reaction times of each individual step may vary depending on the particular reactant used and the substituents present in the reactant used. The reaction may be carried out in a conventional manner, for example by removing the solvent from the residue, and further purified according to methods generally known in the art, such as, but not limited to, crystallization, distillation, extraction, wet milling, and chromatography. Unless otherwise indicated, starting materials and reagents are commercially available or can be prepared from commercially available materials by one skilled in the art using methods described in the chemical literature.

Routine experimentation, including appropriate manipulation of reaction conditions, reagents, and synthetic route sequences, protection of any chemical functionality that is incompatible with the reaction conditions, and deprotection at appropriate points in the reaction sequence of the process, are included within the scope of the present disclosure. Suitable protecting groups and methods for protecting and deprotecting different substituents using such suitable protecting groups are well known to those skilled in the art, examples of which can be found in PGM Wuts and TW Greene, in Greene's book titled Protective Groups in Organic Synthesis (4 th edition), john Wiley & Sons, NY (2006).

When an optically active form of the disclosed compounds is desired, it can be obtained by performing one of the procedures described herein using optically active starting materials (e.g., asymmetric induction preparation by suitable reaction steps) or by resolution of a mixture of stereoisomers of the compound or intermediate using standard procedures such as chromatographic separation, recrystallization, or enzymatic resolution.

Similarly, when a pure geometric isomer of a compound is desired, it may be obtained by performing one of the procedures described herein using the pure geometric isomer as starting material, or by resolving a mixture of geometric isomers of the compound or intermediate using standard procedures (such as chromatographic separation).

The described synthetic schemes and specific examples are illustrative and should not be construed as limiting the scope of the disclosure or claims. Alternatives, modifications, and equivalents of the synthetic methods and specific examples are contemplated.

Pharmaceutical composition

The disclosed compounds can be incorporated into pharmaceutical compositions suitable for administration to a subject (such as a patient, which can be human or non-human). The pharmaceutical composition may comprise a "therapeutically effective amount" or a "prophylactically effective amount" of the agent. "therapeutically effective amount" means an amount effective to achieve the desired therapeutic result at the necessary dosage and time period. The therapeutically effective amount of the composition can be determined by one of skill in the art and can vary depending on factors such as the disease state, age, sex, and weight of the individual, and the ability of the composition to elicit a desired response in the individual. A therapeutically effective amount is also an amount in which any toxic or detrimental effects of the compounds of the present disclosure are exceeded by the therapeutically beneficial effects. "prophylactically effective amount" means an amount effective to achieve the desired prophylactic effect at the dosages and for periods of time necessary. Typically, since a prophylactic dose is administered to a subject prior to or at an early stage of a disease or disorder, the prophylactically effective amount will be less than the therapeutically effective amount.

The pharmaceutical composition may comprise a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" as used herein refers to a non-toxic, inert solid, semi-solid or liquid filler, diluent, encapsulating material or any type of formulation aid. Some examples of materials that may be pharmaceutically acceptable carriers are sugars such as, but not limited to lactose, dextrose, and sucrose, starches such as, but not limited to, corn starch and potato starch, celluloses and derivatives thereof such as, but not limited to, sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate, tragacanth, malt, gelatin, talc, excipients such as, but not limited to, cocoa butter and suppository waxes, oils such as, but not limited to, peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil, glycols such as propylene glycol, esters such as, but not limited to, ethyl oleate and ethyl laurate, agar, buffers such as, but not limited to, magnesium hydroxide and aluminum hydroxide, alginic acid, athermal, isotonic saline, ringer's solution, ethanol and phosphate buffered solutions, and other non-toxic lubricants such as, but not limited to, sodium dodecyl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening agents, flavoring agents and flavoring agents, preservatives and antioxidants may also be present in the composition at the discretion of the formulator.

Thus, the compounds and pharmaceutically acceptable salts thereof may be formulated for administration by, for example, solid administration, eye drops, topical oil-based formulations, injection, inhalation (through the mouth or nose), implants, or oral, buccal, parenteral or rectal administration. Techniques and formulations can generally be found in Remington's Pharmaceutical Sciences (Meade Publishing co., pennsylvania). Therapeutic compositions must generally be sterile and stable under the conditions of manufacture and storage.

The route of administration of the disclosed compounds and the form of the composition will determine the type of carrier to be used. The composition may be in a variety of forms, such as suitable for systemic administration (e.g., oral, rectal, nasal, sublingual, buccal, implant, or parenteral) or topical administration (e.g., dermal, pulmonary, nasal, otic, ocular, liposomal delivery system, or iontophoresis).

Carriers for systemic administration generally include at least one of diluents, lubricants, binders, disintegrants, colorants, flavorants, sweeteners, antioxidants, preservatives, glidants, solvents, suspending agents, wetting agents, surfactants, combinations thereof, and the like. All carriers are optional in the composition.

Suitable diluents include sugars such as glucose, lactose, dextrose, and sucrose, glycols such as propylene glycol, calcium carbonate, sodium carbonate, sugar alcohols such as glycerol, mannitol, and sorbitol. The amount of one or more diluents in the systemic or topical compositions is typically from about 50% to about 90% by weight of the composition.

Suitable lubricants include silica, talc, stearic acid and its magnesium and calcium salts, calcium sulfate, and liquid lubricants such as polyethylene glycol and vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil, and cocoa butter. The amount of one or more lubricants in the systemic or topical composition is typically from about 5% to about 10% by weight of the composition.

Suitable binders include polyvinylpyrrolidone, magnesium aluminum silicate, starches such as corn starch and potato starch, gelatin, tragacanth, and celluloses and derivatives thereof such as sodium carboxymethyl cellulose, ethyl cellulose, methyl cellulose, microcrystalline cellulose and sodium carboxymethyl cellulose. The amount of the one or more binders in the systemic composition is typically from about 5% to about 50% by weight of the composition.

Suitable disintegrants include agar, alginic acid and its sodium salt, effervescent mixtures, croscarmellose, crospovidone, sodium carboxymethyl starch, sodium starch glycolate, clays and ion exchange resins. The amount of one or more disintegrants in a systemic or topical composition is typically from about 0.1% to about 10% by weight of the composition.

Suitable colorants include colorants such as FD & C dyes. When used, the amount of colorant in the systemic or topical composition is typically from about 0.005% to about 0.1% by weight of the composition.

Suitable flavors include menthol, peppermint, and fruit flavors. When used, the amount of one or more fragrances in the systemic or topical compositions is typically from about 0.1% to about 1.0%.

Suitable sweeteners include aspartame and saccharin. When used, the amount of one or more sweeteners in the systemic or topical composition is typically from about 0.001% to about 1% by weight of the composition.

Suitable antioxidants include butylated hydroxyanisole ("BHA"), butylated hydroxytoluene ("BHT"), and vitamin E. The amount of one or more antioxidants in the systemic or topical composition is typically from about 0.1% to about 5% by weight of the composition.

Suitable preservatives include benzalkonium chloride, methylparaben and sodium benzoate. The amount of one or more preservatives in the systemic or topical composition is typically from about 0.01% to about 5% by weight of the composition.

Suitable glidants include silicon dioxide. The amount of one or more glidants in a systemic or topical composition is generally from about 1% to about 5% by weight of the composition.

Suitable solvents include water, isotonic saline, ethyl oleate, glycerol, hydroxylated castor oil, alcohols (e.g., ethanol), and phosphate buffer solutions. The amount of the one or more solvents in the systemic or topical composition is typically from about 0 to about 100% by weight of the composition.

Suitable suspending agents include AVICEL RC-591 (available from FMC Corporation, philadelphia, pa.) and sodium alginate. The amount of one or more suspending agents in the systemic or topical composition is typically from about 1% to about 8% by weight of the composition.

Suitable surfactants include lecithin, polysorbate 80 and sodium lauryl sulfate, as well as tween from Atlas Powder Company of wilmington, tela. Suitable surfactants include those disclosed in C.T.F.A. Cosmetic Ingredient Handbook, 1992, pages 587-592, remington's Pharmaceutical Sciences, 15 th edition 1975, pages 335-337, and McCutcheon, volume 1, emulsifiers & Detergents, 1994, north America, pages 236-239. The amount of one or more surfactants in the systemic or topical compositions is typically from about 0.1% to about 5% by weight of the composition.

Although the amount of components in the systemic composition may vary depending on the type of systemic composition being prepared, in general, the systemic composition comprises from 0.01% to 50% by weight of the active compound and from 50% to 99.99% by weight of one or more carriers. Compositions for parenteral administration typically comprise from 0.1% to 10% by weight of the active substance and from 90% to 99.9% by weight of the carrier, including diluents and solvents.

Compositions for oral administration may have a variety of dosage forms. For example, solid forms include tablets, capsules, granules, and bulk powders. These oral dosage forms comprise a safe and effective amount, typically at least about 5% by weight, more particularly about 25% to about 50% by weight of the active agent. The oral dosage form composition includes from about 50% to about 95% by weight of the carrier, more particularly from about 50% to about 75% by weight.

The tablets may be compressed, tablet ground, enteric coated, sugar coated, film coated or multiply compressed. Tablets typically contain the active ingredient in combination with a carrier containing an ingredient selected from the group consisting of diluents, lubricants, binders, disintegrants, coloring agents, flavoring agents, sweeteners, glidants, and combinations thereof. Specific diluents include calcium carbonate, sodium carbonate, mannitol, lactose and cellulose. Specific binders include starch, gelatin, and sucrose. Specific disintegrants include alginic acid and croscarmellose. Specific lubricants include magnesium stearate, stearic acid and talc. The specific colorant is FD & C dye, which may be added for appearance. The chewable tablet preferably contains a sweetener such as aspartame and saccharin, or a flavoring such as menthol, peppermint, fruit flavors, or combinations thereof.

Capsules (including implants, timed release and sustained release formulations) typically include an active compound (e.g., a compound of formula (I)) and a carrier comprising one or more of the diluents disclosed above in a capsule containing gelatin. The particles typically contain the disclosed compounds, and preferably a glidant such as silicon dioxide to improve flow characteristics. The implant may be of the biodegradable or non-biodegradable type.

The choice of ingredients in the carrier of the oral composition depends on secondary considerations such as taste, cost and shelf stability, which are not important for the purposes of the present disclosure.

The solid compositions may be coated by conventional methods, typically with a pH or time dependent coating, such that the disclosed compounds are released in the gastrointestinal tract near the desired application, or at different points and times, to prolong the desired effect. The coating typically comprises one or more components selected from the group consisting of cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl methyl cellulose phthalate, ethyl cellulose, EUDRAGIT (available from Evonik Industries of Essen, germany), waxes and shellac.

Compositions for oral administration may have a liquid form. For example, suitable liquid forms include aqueous solutions, emulsions, suspensions, solutions reconstituted with non-effervescent granules, suspensions reconstituted with non-effervescent granules, effervescent formulations reconstituted with effervescent granules, elixirs, tinctures, syrups, and the like. Liquid oral compositions typically comprise a disclosed compound and a carrier, i.e., a carrier selected from diluents, colorants, flavors, sweeteners, preservatives, solvents, suspending agents, and surfactants. The oral liquid composition preferably comprises one or more ingredients selected from the group consisting of colorants, flavors and sweeteners.

Other compositions that may be used to achieve systemic delivery of the subject compounds include sublingual, buccal and nasal dosage forms. Such compositions typically comprise one or more soluble filler materials, such as diluents, including sucrose, sorbitol and mannitol, and binders, such as acacia, microcrystalline cellulose, carboxymethylcellulose and hydroxypropyl methylcellulose. Such compositions may also contain lubricants, colorants, flavors, sweeteners, antioxidants, and glidants.

The disclosed compounds may be administered topically. Topical compositions that may be topically applied to the skin may be in any form, including solids, solutions, oils, creams, ointments, gels, emulsions, shampoos, leave-in and rinse-off conditioners, emulsions, cleansers, moisturizers, sprays, skin patches, and the like. The topical compositions comprise a disclosed compound (e.g., a compound of formula (I)) or a pharmaceutically acceptable salt thereof and a carrier. The carrier of the topical composition preferably assists penetration of the compound into the skin. The carrier may also include one or more optional components.

The amount of carrier used in combination with the disclosed compounds is sufficient to provide a practical amount of composition for administration per unit dose of the compound. Techniques and compositions for making dosage forms useful in the methods of the present disclosure are described in the following references Modern Pharmaceutics, chapter 9 and 10, banker & Rhodes (1979), lieberman et al Pharmaceutical Dosage Forms: tablets (1981), and Ansel, introduction to Pharmaceutical Dosage Forms, 2 nd edition (1976).

The carrier may comprise a single ingredient or a combination of two or more ingredients. In the topical compositions, the carrier comprises a topical carrier. Suitable topical carriers include one or more ingredients selected from phosphate buffered saline, isotonic water, deionized water, monofunctional alcohols, symmetrical alcohols, aloe vera gel, allantoin, glycerin, vitamin a and E oils, mineral oil, propylene glycol, PPG-2 myristyl propionate, dimethyl isosorbide, castor oil, combinations thereof, and the like. More particularly, carriers for skin applications include propylene glycol, dimethyl isosorbide, and water, even more particularly, phosphate buffered saline, isotonic water, deionized water, monofunctional alcohols, and symmetrical alcohols.

The carrier of the topical composition may also include one or more ingredients selected from the group consisting of emollients, propellants, solvents, moisturizers, thickeners, powders, fragrances, pigments, and preservatives, all of which are optional.

Suitable emollients include stearyl alcohol, glycerol monoricinoleate, glycerol monostearate, propane-1, 2-diol, butane-1, 3-diol, mink oil, cetyl alcohol, isopropyl isostearate, stearic acid, isobutyl palmitate, isocetyl stearate, oleyl alcohol, isopropyl laurate, hexyl laurate, decyl oleate, stearyl-2-ol, isocetyl alcohol, cetyl palmitate, di-n-butyl sebacate, isopropyl myristate, isopropyl palmitate, isopropyl stearate, butyl stearate, polyethylene glycol, triethylene glycol, lanolin, sesame oil, coconut oil, peanut oil, castor oil, acetylated lanolin alcohol, petroleum, mineral oil, butyl myristate, isostearic acid, palmitic acid, isopropyl linoleate, lauryl lactate, myristyl lactate, decyl oleate, myristyl myristate, and combinations thereof. Specific emollients for the skin include stearyl alcohol and polydimethylsiloxane. The amount of one or more emollients in a skin-based topical composition is typically from about 5% to about 95% by weight of the composition.

Suitable propellants include propane, butane, isobutane, dimethyl ether, carbon dioxide, nitrous oxide, and combinations thereof. The amount of the one or more propellants in the topical composition is generally from about 0% to about 95% by weight of the composition.

Suitable solvents include water, ethanol, methylene chloride, isopropyl alcohol, castor oil, ethylene glycol monoethyl ether, diethylene glycol monobutyl ether, diethylene glycol monoethyl ether, dimethyl sulfoxide, dimethylformamide, tetrahydrofuran, and combinations thereof. Specific solvents include ethanol and isopolyols. The amount of the one or more solvents in the topical composition is typically from about 0% to about 95% by weight of the composition.

Suitable humectants include glycerin, sorbitol, sodium 2-pyrrolidone-5-carboxylate, soluble collagen, dibutyl phthalate, gelatin, and combinations thereof. Specific humectants include glycerin. The amount of one or more humectants in a topical composition is typically from 0% to 95% by weight of the composition.

The amount of one or more thickeners in the topical composition is typically from about 0% to about 95% by weight of the composition.

Suitable powders include beta-cyclodextrin, hydroxypropyl cyclodextrin, chalk, talc, fullerenes, kaolin, starch, gums, colloidal silica, sodium polyacrylate, tetraalkyl montmorillonite ammonium, trialkyl aryl montmorillonite ammonium, chemically modified magnesium aluminum silicate, organically modified montmorillonite clay, hydrated aluminum silicate, fumed silica, carboxyvinyl polymer, sodium carboxymethyl cellulose, ethylene glycol monostearate, and combinations thereof. The amount of the one or more powders in the topical composition is typically from 0% to 95% by weight of the composition.

The amount of perfume in the topical composition is generally from about 0% to about 0.5%, particularly from about 0.001% to about 0.1% by weight of the composition.

Suitable pH adjusting additives include HCl or NaOH in an amount sufficient to adjust the pH of the topical pharmaceutical composition.

Application method

The disclosed compounds and pharmaceutical compositions may be used in methods for treating conditions, including proliferative conditions (such as cancer). In some embodiments, the cancer is characterized by or is mediated by BCL6 activity. In some embodiments, the cancer is a cancer in which BCL6 and p300/CBP are expressed.

Thus, in some embodiments, disclosed herein is a method of treating a disorder in a subject in need thereof, wherein the disorder is characterized by or mediated by BCL6 activity, or a cancer in which BCL6 and p300/CBP are expressed, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein (e.g., a compound of formula (I) or a pharmaceutically acceptable salt thereof), or a pharmaceutical composition comprising a compound disclosed herein (e.g., a compound of formula (I) or a pharmaceutically acceptable salt thereof). In some embodiments, the disorder is a proliferative disease, i.e., a disease that occurs due to abnormal growth or expansion caused by cell multiplication. In some embodiments, the proliferative disease is cancer. The term "cancer" refers to a class of diseases characterized by uncontrolled proliferation of abnormal cell development and the ability to penetrate and destroy normal body tissues. See, e.g., stedman's Medical Dictionary, 25 th edition, volume Hensyl, williams & Wilkins: philadelphia, 1990.

In some embodiments, the compounds and pharmaceutical compositions disclosed herein are used to treat cancer in a patient in need thereof. In some embodiments, the cancer is a cancer in which BCL6 is mutated. In some embodiments, the cancer is a cancer in which BCL6 is highly expressed, or overexpressed relative to non-cancer cells.

In some embodiments, the cancer is a solid tumor, such as a sarcoma or carcinoma. In some embodiments, the cancer is a hematological malignancy.

Exemplary sarcomas include, but are not limited to alveolar rhabdomyosarcoma, alveolar soft tissue sarcoma, amelogenetic carcinoma, angiosarcoma, chondrosarcoma, chordoma, parenchymal clear cell sarcoma, dedifferentiated liposarcoma, hard fibrosarcoma, desmoplasia small round cell tumor, embryonal rhabdomyosarcoma, epithelioid fibrosarcoma, epithelioid endothelioma, olfactory neuroblastoma, ewing's sarcoma, renal exorhabdoid sarcoma, osteo-mucoid chondrosarcoma, osteo-osteosarcoma, fibrosarcoma, giant cell tumor, vascular epidermoid carcinoma, infant fibrosarcoma, inflammatory myofibroblastoma, kaposi's sarcoma, osteoleiomyosarcoma, liposarcoma, psoralen-fatty sarcoma, malignant Fibrous Histiocytoma (MFH), osteomalignant fibrous histiocytoma (MFH), malignant mesenchymal tumor, malignant peripheral nerve sheath tumor, mesenchymal chondrosarcoma, myxofibrosarcoma, myxoid liposarcoma, myxoid fibrosarcoma, tumors with perivascular epithelial differentiation, osteosarcoma with perivascular differentiation, peri-myxomatoid osteosarcoma, peri-myxomatoid sarcoma, and myxomatoid sarcoma, angiosarcoma, angio-tumor, osteosarcoma, peri-tumor, osteosarcoma, and myxomatoid sarcoma, and myxoma.

Exemplary cancers include, but are not limited to, adenocarcinoma, squamous cell carcinoma, adenosquamous carcinoma, undifferentiated carcinoma, large cell carcinoma, small cell carcinoma, anal carcinoma, appendiceal carcinoma, cholangiocarcinoma (i.e., bile duct carcinoma (cholangiocarcinoma)), bladder carcinoma, brain tumor, breast carcinoma, cervical carcinoma, colon carcinoma, primary Carcinoma (CUP), esophageal carcinoma (e.g., esophageal squamous cell carcinoma), eye carcinoma, fallopian tube carcinoma, gastrointestinal carcinoma, renal carcinoma, liver carcinoma (e.g., hepatocellular carcinoma), lung carcinoma, myeloblastoma, melanoma, oral carcinoma, ovarian carcinoma, pancreatic carcinoma, parathyroid disease, penile carcinoma, pituitary carcinoma, prostate carcinoma, rectal carcinoma, skin carcinoma, gastric carcinoma, testicular carcinoma, throat carcinoma, thyroid carcinoma, uterine carcinoma, vaginal carcinoma, and vulvar carcinoma.

Exemplary hematological malignancies include, but are not limited to, leukemia, lymphoma, myeloma, non-hodgkin's lymphoma, T-cell malignancy, and B-cell malignancy. Exemplary T cell malignancies include anaplastic large cell lymphoma, angioimmunoblastic lymphoma, adult T cell leukemia/lymphoma (ATLL), maternal NK cell lymphoma, cutaneous T cell lymphoma, enteropathy type T cell lymphoma, blood spleen γδ T cell lymphoma, lymphoblastic lymphoma, nasal NK/T cell lymphoma, peripheral T cell lymphoma not otherwise specified (PTCL-NOS), and treatment-related T cell lymphoma. Exemplary B-cell malignancies include Chronic Lymphocytic Leukemia (CLL), small Lymphocytic Lymphomas (SLL), high-risk CLL, and non-CLL/SLL lymphomas. In some embodiments, the cancer is selected from the group consisting of B-cell pre-lymphocytic leukemia, burkitt's lymphoma, diffuse large B-cell lymphoma (DLBCL), extranodal peripheral zone B-cell lymphoma, follicular Lymphoma (FL), immunoblastic large cell lymphoma, intravascular large B-cell lymphoma, lymphomatoid granuloma, lymphoplasmacytic lymphoma, mantle Cell Lymphoma (MCL), mediastinal (thymic) large B-cell lymphoma, multiple myeloma, extranodal peripheral zone B-cell lymphoma, non-burkitt's high grade B-cell lymphoma, plasma cell myeloma, plasma cell lymphoma, precursor B-lymphoblastic lymphoma, primary exudative lymphoma, primary mediastinal B-cell lymphoma (PMBL), splenic peripheral zone lymphoma, or fahrenheit macroglobulinemia. In some embodiments, the cancer is diffuse large B-cell lymphoma (DLBCL).

In some embodiments, the cancer is a recurrent or refractory cancer, such as the cancers described herein. In some embodiments, the cancer is a metastatic cancer, such as the cancers described herein.

In the methods of treatment disclosed herein, the compound or pharmaceutical composition may be administered to a subject by any convenient route of administration, whether systemic/peripheral or at the desired site of action, including but not limited to oral (e.g., by ingestion), topical (including, e.g., transdermal, intranasal, ocular, buccal, and sublingual), pulmonary (e.g., by inhalation or insufflation using, e.g., an aerosol, e.g., through the mouth or nose), rectal, vaginal, parenteral (e.g., by injection, including subcutaneous, intradermal, intramuscular, intravenous, intra-arterial, intracardiac, intrathecal, intraspinal, intracapsular, intraorbital, intraperitoneal, intratracheal, subcutical, intra-articular, subarachnoid, and intrasternal), or by implantation of a depot, e.g., subcutaneous or intramuscular. In some embodiments, the administering comprises oral administration. In some embodiments, the administering comprises parenteral administration. Additional modes of administration may include adding the compound and/or a composition comprising the compound to food or beverage, including drinking water for animals, to provide the compound as part of the animal's diet.

It will be appreciated that the appropriate dosage of the compounds and compositions comprising the compounds may vary from patient to patient. Determining the optimal dose will generally involve balancing the level of therapeutic benefit against any risk or deleterious side effects of the treatment of the present disclosure. The selected dosage level will depend on a variety of factors including, but not limited to, the activity of the particular compound, the route of administration, the time of administration, the rate of excretion of the compound, the duration of the treatment, other drugs, compounds and/or materials used in combination, as well as the age, sex, weight, condition, general health and prior medical history of the patient. The amount of the compound and the route of administration will ultimately be at the discretion of the physician, but generally the dosage will result in a local concentration at the site of action which achieves the desired effect without causing generally deleterious or toxic side effects.

In vivo administration may be achieved in one dose, continuously or intermittently (e.g., in divided doses at appropriate intervals) throughout the course of treatment. Methods of determining the most effective mode and dosage of administration are well known to those skilled in the art and will vary with the formulation used for the treatment, the purpose of the treatment, the target cells being treated and the subject being treated. Single or multiple administrations may be performed depending on the dosage level and mode selected by the treating physician. Generally, a suitable dose of the compound is in the range of about 100 μg/kg subject weight to about 250 mg/kg subject weight per day.

The compound or composition may be administered once, or may be administered continuously (e.g., by intravenous drip), or may be administered periodically/intermittently, including about once every hour, about once every two hours, about once every four hours, about once every eight hours, about once every twelve hours, about once every day, about once every two days, about once every three days, about twice weekly, about once weekly, and about once monthly. The composition may be administered until the desired relief of symptoms is achieved.

The compounds described herein may be used in combination with other known therapies. "combined" administration as used herein means that two (or more) different treatments are delivered to a subject during the course of the subject suffering from a disorder, e.g., after the subject is diagnosed with a disorder, and before the disorder is cured or eliminated or treatment is stopped for other reasons. In some embodiments, when delivery of the second treatment begins, delivery of one treatment is still in progress, so there is overlap in administration. This is sometimes referred to herein as "simultaneous delivery" or "parallel delivery. In other embodiments, the delivery of one therapy ends before the delivery of another therapy begins. In some embodiments of either case, the treatment is more effective due to the combined administration. For example, the second treatment is more effective than that observed when the second treatment is administered without the first treatment, e.g., the same effect is observed with less of the second treatment, or the second treatment reduces symptoms to a greater extent, or similar conditions are observed with the first treatment. In some embodiments, the delivery is such that the reduction in symptoms or other parameters associated with the disorder is greater than that observed when one treatment is delivered without another treatment. The effect of both treatments may be partially additive, fully additive, or more than additive. The delivery may be such that the effect of the first treatment is still detectable when the second treatment is delivered.

The compounds or compositions described herein may be administered simultaneously, in the same or separate compositions, or sequentially with at least one additional therapeutic agent. For sequential administration, the compounds described herein may be administered first, and additional agents may be administered subsequently, or the order of administration may be reversed.

In some embodiments, the compounds described herein are administered in combination with other therapeutic modalities including surgery, radiation, transplantation (e.g., stem cell transplantation, bone marrow transplantation), cryotherapy, and/or thermal therapy. Such combination therapies may allow for lower doses of administered agents and/or other chemotherapeutic agents, thereby avoiding toxicity or complications associated with the various therapies.

In some embodiments, the compounds described herein are administered with at least one additional therapeutic agent (such as a chemotherapeutic agent). In certain embodiments, the compounds described herein are administered in combination with one or more additional chemotherapeutic agents. The chemotherapeutic agent may be one identified in the "cancer drugs a to Z list" issued by the national cancer institute.

Also disclosed herein are methods of reducing proliferation of a cancer cell in a sample, comprising contacting the sample with a compound described herein (e.g., a compound of formula (I) or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition described herein.

Medicine box

The compounds and/or compositions disclosed herein may be assembled into a kit or pharmaceutical system. The kit or pharmaceutical system may comprise a carrier or package such as a box, carton, tube, etc., having tightly closed therein one or more containers, such as vials, tubes, ampoules or bottles, containing a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof. The kit or pharmaceutical system may also include printed instructions for using the compounds and/or compositions.

The following examples further illustrate aspects of the disclosure, but should not be construed as limiting its scope in any way.

Examples

The following abbreviations are used in the examples: DIPEA is N, N-diisopropylethylamine, DMF is N, N-dimethylformamide, DMSO is dimethylsulfoxide, HATU is 1- [ bis (dimethylamino) methylene ] -1H-1,2, 3-triazolo [4,5-b ] pyridinium 3-oxide hexafluorophosphate, HPLC is high performance liquid chromatography, TFA is trifluoroacetic acid, and UP-LC is ultra-high performance liquid chromatography.

EXAMPLE 1 Synthesis of Compounds

Procedure A (MNN-02-155) to a solution of 5- ((tert-butoxycarbonyl) amino) pentanoic acid (4.35 mg, 0.0200 mmol, 1.50 eq.) in anhydrous DMF (300 uL, 0.05M) and DIPEA (6.97 uL, 0.0400 mmol, 3.00 eq.) under nitrogen was added HATU (7.61 mg, mmol, 1.50 eq.). The reaction was stirred at 0 ℃ for 15 min, and then 3- (7- (difluoromethyl) -6- (1-methyl-1H-pyrazol-4-yl) -3, 4-dihydroquinolin-1 (2H) -yl) -N-methyl-1- (piperidin-4-yl) -1,4,6, 7-tetrahydro-5H-pyrazolo [4,3-C ] pyridine-5-carboxamide (7.00 mg, 0.0133 mmol,1.00 eq) (1 mg/10 μl) dissolved in DMSO was added dropwise. The reaction was stirred for 3 hours, at which point the product mass (MS M/z 724.62 [ M+H ] ⁺) of acceptable purity was observed by UP-LC. The reaction was diluted in water and the product extracted 3 times in ethyl acetate, dried over Na ₂SO₄ and concentrated in vacuo. The crude product was then dissolved in 500 uL dichloromethane and 100 μl TFA and stirred at room temperature for 1 hour, at which point the solution was concentrated in vacuo, azeotroped 3 times with dichloromethane and left under vacuum for 12 hours.

Procedure B (MNN-02-155) DIPEA (11.7 μl, 0.0673 mmol, 7.00 eq) and HATU (5.49 mg, 0.0144 mmol, 1.50 eq) were added to a solution of 1- (5-chloro-4- ((8-methoxy-1-methyl-3- (2- (methylamino) -2-oxoethoxy) -2-oxo-1, 2-dihydroquinolin-6-yl) amino) pyrimidin-2-yl) piperidine-4-carboxylic acid (7.66 mg, 0.0144 mmol, 1.50 eq) in DMF (200 μl, 0.05M) and the mixture was stirred under nitrogen at 0 ℃ for 15 min. The product of procedure a (6.00 mg, 0.00962 mmol, 1.00 eq.) dissolved in 200 μl DMF was then added dropwise and the mixture stirred for 12 hours. The reaction mixture was directly purified by preparative HPLC (85% -20% water (0.05% TFA)/methanol). The product 1- (1- (5-chloro-4- ((8-methoxy-1-methyl-3- (2- (methylamino) -2-oxoethoxy) -2-oxo-1, 2-dihydroquinolin-6-yl) amino) pyrimidin-2-yl) piperidine-4-carboxamido) pentanoyl) piperidin-4-yl) -3- (7- (difluoromethyl) -6- (1-methyl-1H-pyrazol-4-yl) -3, 4-dihydroquinolin-1 (2H) -yl) -N-methyl-1, 4,6, 7-tetrahydro-5H-pyrazolo [4,3-c ] pyridine-5-carboxamide was obtained as a white solid (4.28 mg, 40%, MS m/z 1136.52 [M+H]⁺).¹H NMR (500 MHz, DMSO-d⁶) δ = 9.03 (s, 1H), 8.10 (d, J = 6.0 Hz, 1H), 7.95 (q, J = 4.5 Hz, 1H), 7.79 (t, J = 5.6 Hz, 1H), 7.73 (s, 1H), 7.51 (s, 2H), 7.48 (s, 1H), 7.08 (s, 1H), 7.01 (s, 1H), 6.78 (s, 1H), 6.77 (t, J = 55.3 Hz, 1H), 6.54 (s, 1H), 4.55 (s, 2H), 4.49 – 4.43 (m, 3H), 4.31 (td, J = 10.6, 5.4 Hz, 1H), 4.01 (s, 2H), 3.95 (d, J = 13.7 Hz, 1H), 3.87 (d, J = 3.4 Hz, 3H), 3.85 (s, 3H), 3.85 (s, 3H), 3.60 (q, J = 6.1 Hz, 2H), 3.56 (t, J = 5.6 Hz, 2H), 3.21 – 3.09 (m, 1H), 3.03 (q, J = 7.0 Hz, 2H), 2.95 – 2.87 (m, 2H), 2.82 (t, J = 6.3 Hz, 2H), 2.76 – 2.71 (m, 2H), 2.68 (d, J = 12.5 Hz, 1H), 2.64 (d, J = 4.7 Hz, 3H), 2.54 (s, 3H), 2.42 – 2.35 (m, 1H), 2.33 (t, J = 7.3 Hz, 2H), 1.96 (q, J = 5.9 Hz, 2H), 1.91 – 1.82 (m, 3H), 1.70 (d, J = 13.1 Hz, 3H), 1.58-1.43 (m, 4H), 1.40 (t, J = 7.4 Hz, 2H).

MNN-03-037 (1- (1- (4- (1- (5-chloro-4- ((8-methoxy-1-methyl-3- (2- (methylamino) -2-oxoethoxy) -2-oxo-1, 2-dihydroquinolin-6-yl) amino) pyrimidin-2-yl) piperidine-4-carboxamido) butyryl) piperidin-4-yl) -3- (7- (difluoromethyl) -6- (1-methyl-1H-pyrazol-4-yl) -3, 4-dihydroquinolin-1 (2H) -yl) -N-methyl-1, 4,6, 7-tetrahydro-5H-pyrazolo [4,3-c ] pyridine-5-carboxamide). Synthesis of Compound MNN-03-037 using linker 4- ((tert-butoxycarbonyl) amino) butanoic acid according to procedure A, and subsequent passage through procedure B, gives the title compound (1.95 mg, 18%, MS m/z 1122.51 [M+H]⁺).¹H NMR (500 MHz, DMSO-d⁶) δ = 9.03 (s, 1H), 8.09 (s, 1H), 7.97 – 7.91 (m, 1H), 7.81 (t, J = 5.7 Hz, 1H), 7.74 (d, J = 2.6 Hz, 1H), 7.51 (d, J = 2.7 Hz, 2H), 7.49 (s, 1H), 7.08 (s, 1H), 7.00 (s, 1H), 6.78 (s, 1H), 6.77 (t, J = 55.3 Hz, 1H), 6.54 (s, 1H), 4.55 (s, 2H), 4.46 (d, J = 12.9 Hz, 3H), 4.35 – 4.26 (m, 1H), 4.01 (s, 2H), 3.93 (d, J = 13.4 Hz, 1H), 3.86 (s, 3H), 3.85 (s, 3H), 3.85 (s, 3H), 3.66-3.58 (m, 2H), 3.56 (t, J = 5.7 Hz, 2H), 3.14 (d, J = 12.3 Hz, 1H), 3.05 (q, J = 6.6 Hz, 2H), 2.96 – 2.87 (m, 2H), 2.81 (d, J = 6.5 Hz, 2H), 2.73 (d, J = 5.5 Hz, 2H), 2.69 (d, J = 12.7 Hz, 1H), 2.64 (d, J = 4.7 Hz, 3H), 2.54 (s, 3H), 2.42 – 2.36 (m, 1H), 2.33 (t, J = 7.5 Hz, 2H), 1.95 (p, J = 6.3 Hz, 2H), 1.87 (d, J = 17.1 Hz, 3H), 1.75 – 1.68 (m, 3H), 1.62 (p, J = 7.2 Hz, 2H), 1.50 (td, J = 12.2, 8.6 Hz, 2H).

MNN-02-195- (1- (6- (1- (5-chloro-4- ((8-methoxy-1-methyl-3- (2- (methylamino) -2-oxoethoxy) -2-oxo-1, 2-dihydroquinolin-6-yl) amino) pyrimidin-2-yl) piperidine-4-carboxamido) hexan-4-yl) -piperidin-4-yl) -3- (7- (difluoromethyl) -6- (1-methyl-1H-pyrazol-4-yl) -3, 4-dihydroquinolin-1 (2H) -yl) -N-methyl-1, 4,6, 7-tetrahydro-5H-pyrazolo [4,3-c ] pyridine-5-carboxamide. Compound MNN-02-195 was synthesized according to procedure A using linker 6- ((tert-butoxycarbonyl) amino) hexanoic acid, and then passed through procedure B to give the title compound (3.52 mg, 20%, MS m/z 1150.53 [M+H]⁺).¹H NMR (500 MHz, DMSO-d⁶) δ = 9.12 (s, 1H), 8.12 (d, J = 5.5 Hz, 1H), 7.95 (q, J = 4.5 Hz, 1H), 7.77 (t, J = 5.6 Hz, 1H), 7.73 (s, 1H), 7.51 (s, 2H), 7.48 (s, 1H), 7.08 (s, 1H), 7.01 (s, 1H), 6.78 (s, 1H), 6.77 (t, J = 55.2 Hz, 1H), 6.54 (s, 1H), 4.55 (s, 2H), 4.44 (d, J = 13.2 Hz, 3H), 4.36 – 4.26 (m, 1H), 4.01 (s, 2H), 3.95 (d, J = 13.2 Hz, 1H), 3.86 (s, 3H), 3.85 (s, 3H), 3.85 (s, 3H), 3.60 (q, J = 5.8 Hz, 2H), 3.56 (t, J = 5.6 Hz, 2H), 3.15 (t, J = 12.0 Hz, 1H), 3.01 (q, J = 6.8 Hz, 2H), 2.93 (t, J= 12.5 Hz, 2H), 2.82 (t, J = 6.5 Hz, 2H), 2.73 (t, J = 5.9 Hz, 2H), 2.71 – 2.66 (m, 1H), 2.64 (d, J = 4.6 Hz, 3H), 2.54 (s, 3H), 2.41 – 2.34 (m, 1H), 2.31 (t, J = 7.5 Hz, 2H), 1.95 (t, J = 5.8 Hz, 2H), 1.91 – 1.83 (m, 3H), 1.71 (dd, J = 13.2, 3.7 Hz, 3H), 1.49 (dt, J = 14.5, 5.6 Hz, 4H), 1.39 (p, J = 7.1 Hz, 2H), 1.25 (qd, J = 8.5, 5.8 Hz, 2H).

RCS-SBM-IJD-001:1- (1- (7- (1- (5-chloro-4- ((8-methoxy-1-methyl-3- (2- (methylamino) -2-oxoethoxy) -2-oxo-1, 2-dihydroquinolin-6-yl) amino) pyrimidin-2-yl) piperidine-4-carboxamido) heptanoyl) piperidin-4-yl) -3- (7- (difluoromethyl) -6- (1-methyl-1H-pyrazol-4-yl) -3, 4-dihydroquinolin-1 (2H) -yl) -N-methyl-1, 4,6, 7-tetrahydro-5H-pyrazolo [4,3-c ] pyridine-5-carboxamide. Synthesis of Compound RCS-SBM-IJD-001 using linker 8- ((tert-butoxycarbonyl) amino) octanoic acid according to procedure A, and subsequent passage through procedure B to give title compound (3.92 mg, 18%, MS m/z 1165.48 [M+H]⁺).¹H NMR (500 MHz, DMSO-d⁶) δ = 9.16 (s, 1H), 8.12 (s, 1H), 7.95 (q, J = 4.6 Hz, 1H), 7.76 (t, J = 5.6 Hz, 1H), 7.73 (s, 1H), 7.50 (s, 2H), 7.48 (d, J = 0.8 Hz, 1H), 7.08 (s, 1H), 7.01 (s, 1H), 6.79 (s, 1H), 6.77 (t, J = 55.2 Hz, 1H), 6.53 (s, 1H), 4.55 (s, 2H, attributed to HSQC, 4.46-4.40 (3H, attributed to HSQC), 4.35-4.27 (m, 1H, attributed to HSQC) ), 4.01 (s, 2H), 3.95 (d, J = 13.6 Hz, 1H), 3.86 (s, 3H), 3.86 (s, 3H), 3.85 (s, 3H), 3.60 (q, J = 5.8 Hz, 2H), 3.56 (t, J = 5.7 Hz, 2H), 3.15 (t, J = 12.5 Hz, 1H), 3.00 (q, J = 6.6 Hz, 2H), 2.97 – 2.89 (m, 2H), 2.82 (t, J = 6.3 Hz, 2H), 2.73 (t, J = 6.0 Hz, 2H), 2.68 (d, J = 12.1 Hz, 1H), 2.64 (d, J = 4.6 Hz, 3H), 2.54 (s, 3H), 2.42 – 2.34 (m, 1H), 2.31 (t, J = 7.5 Hz, 2H), 1.96 (p, J = 6.3 Hz, 2H), 1.88 (dd, J = 18.7, 8.7 Hz, 3H), 1.74 – 1.68 (m, 3H), 1.56 – 1.42 (m, 4H), 1.36 (q, J = 7.0 Hz, 2H), 1.25 (d, J = 5.4 Hz, 4H).

MNN-02-196 1- (1- (8- (1- (5-chloro-4- ((8-methoxy-1-methyl-3- (2- (methylamino) -2-oxoethoxy) -2-oxo-1, 2-dihydroquinolin-6-yl) amino) pyrimidin-2-yl) piperidine-4-carboxamido) octanoyl) piperidin-4-yl) -3- (7- (difluoromethyl) -6- (1-methyl-1H-pyrazol-4-yl) -3, 4-dihydroquinolin-1 (2H) -yl) -N-methyl-1, 4,6, 7-tetrahydro-5H-pyrazolo [4,3-c ] pyridine-5-carboxamide. Synthesis of Compound MNN-02-196 using linker 8- ((tert-butoxycarbonyl) amino) octanoic acid according to procedure A and subsequent passage through procedure B to give title compound (1.43 mg, 8%, MS m/z 1180.09 [M+H]⁺).¹H NMR (500 MHz, DMSO-d⁶) δ = 8.86 (d, J = 8.5 Hz, 1H), 8.06 (d, J = 8.2 Hz, 1H), 8.02 – 7.97 (m, 1H), 7.74 (q, J = 7.3 Hz, 2H), 7.52 (d, J = 8.9 Hz, 2H), 7.47 (d, J = 8.9 Hz, 1H), 7.08 (d, J = 8.8 Hz, 1H), 7.00 (d, J = 8.6 Hz, 1H), 6.79 (s, 1H), 6.76 (t, 1H), 6.53 (s, 1H), 4.54 (s, 2H), 4.50 (d, J = 21.7 Hz, 3H), 4.35 – 4.26 (m, 1H), 4.00 (d, J = 8.7 Hz, 2H), 3.94 (d, J = 13.2 Hz, 1H), 3.87 (d, J = 2.2 Hz, 3H), 3.86 – 3.82 (m, 6H), 3.68-3.34 (4H, attributed to HSQC ), 3.14 (q, J = 10.1 Hz, 1H), 3.00 (q, J = 6.7 Hz, 2H), 2.88 (t, J = 12.1 Hz, 2H), 2.82 (t, J = 6.5 Hz, 2H), 2.73 (t, J = 6.1 Hz, 2H), 2.68 (d, J = 12.1 Hz, 1H), 2.64 (dd, J = 9.2, 4.6 Hz, 3H), 2.54 (s, 3H), 2.36 (q, J = 5.2 Hz, 1H), 2.30 (t, J = 7.5 Hz, 2H), 1.95 (q, J = 6.2 Hz, 2H), 1.86 (d, J = 10.5 Hz, 3H), 1.69 (d, J = 12.3 Hz, 3H), 1.47 (t, J = 11.0 Hz, 4H), 1.35 (d, J = 8.0 Hz, 2H), 1.29 – 1.14 (m, 6H).

MNN-02-187 1- (1- (9- (1- (5-chloro-4- ((8-methoxy-1-methyl-3- (2- (methylamino) -2-oxoethoxy) -2-oxo-1, 2-dihydroquinolin-6-yl) amino) pyrimidin-2-yl) piperidine-4-carboxamido) nonanoyl) piperidin-4-yl) -3- (7- (difluoromethyl) -6- (1-methyl-1H-pyrazol-4-yl) -3, 4-dihydroquinolin-1 (2H) -yl) -N-methyl-1, 4,6, 7-tetrahydro-5H-pyrazolo [4,3-c ] pyridine-5-carboxamide. Synthesis of Compound MNN-02-187 using linker 9- ((tert-butoxycarbonyl) amino) nonanoic acid according to procedure A, and subsequent passage through procedure B, gives the title compound (1.41 mg, 8%, MS m/z 1193.80 [M+H]⁺).¹H NMR (500 MHz, DMSO-d⁶) δ = 8.10 (d, J = 3.5 Hz, 1H), 7.95 (q, J = 4.6 Hz, 1H), 7.79 – 7.72 (m, 2H), 7.51 (s, 2H), 7.48 (s, 1H), 7.08 (s, 1H), 7.01 (d, J = 2.6 Hz, 1H), 6.79 (s, 1H), 6.76 (t, J = 55.2 Hz, 1H), 6.54 (s, 1H), 4.54 (s, 2H), 4.46 (d, J = 13.1 Hz, 3H), 4.31 (s, 1H), 4.01 (s, 2H), 3.95 (d, J = 13.7 Hz, 1H), 3.88 – 3.83 (m, 9H), 3.58 (dt, J = 22.9, 5.6 Hz, 4H), 3.19 – 3.11 (m, 1H), 3.09 – 2.97 (m, 2H), 2.97 – 2.87 (m, 2H), 2.82 (t, J = 6.4 Hz, 2H), 2.79 – 2.71 (m, 2H), 2.69 (s, 1H), 2.64 (d, J= 4.7 Hz, 3H), 2.53 (s, 3H), 2.38 (d, J = 11.7 Hz, 2H), 2.30 (t, J = 7.4 Hz, 2H), 1.95 (p, J = 6.0 Hz, 3H), 1.89 (s, 3H), 1.70 (d, J = 13.2 Hz, 3H), 1.56 – 1.42 (m, 4H), 1.38 – 1.28 (m, 2H), 1.28 – 1.20 (m, 6H).

MNN-02-197 1- (1- (10- (1- (5-chloro-4- ((8-methoxy-1-methyl-3- (2- (methylamino) -2-oxoethoxy) -2-oxo-1, 2-dihydroquinolin-6-yl) amino) pyrimidin-2-yl) piperidine-4-carboxamido) decanoyl) piperidin-4-yl) -3- (7- (difluoromethyl) -6- (1-methyl-1H-pyrazol-4-yl) -3, 4-dihydroquinolin-1 (2H) -yl) -N-methyl-1, 4,6, 7-tetrahydro-5H-pyrazolo [4,3-c ] pyridine-5-carboxamide. Compound MNN-02-197 was synthesized according to procedure A using linker 10- ((tert-butoxycarbonyl) amino) decanoic acid, and then passed through procedure B to give the title compound (1.41 mg, 8%, MS m/z 1207.76 [M+H]⁺).¹H NMR (500 MHz, DMSO-d⁶) δ = 9.10 (s, 1H), 8.11 (s, 1H), 7.95 (q, J = 4.6 Hz, 1H), 7.75 (t, J = 5.6 Hz, 1H), 7.74 (s, 1H), 7.51 (s, 2H), 7.48 (d, J = 0.9 Hz, 1H), 7.08 (s, 1H), 7.01 (d, J = 1.8 Hz, 1H), 6.79 (s, 1H), 6.77 (t, J = 55.3 Hz, 1H), 6.54 (s, 1H), 4.55 (s, 2H), 4.45 (d, J = 12.9 Hz, 3H), 4.30 (m, J = 10.4, 5.3 Hz, 1H), 4.01 (s, 2H), 3.95 (d, J = 13.6 Hz, 1H), 3.87 (s, 3H), 3.86 (s, 3H), 3.85 (s, 3H), 3.65 – 3.53 (m, 4H), 3.18 – 3.09 (m, 1H), 2.99 (q, J = 6.6 Hz, 2H), 2.97 – 2.89 (m, 2H), 2.82 (t, J = 6.4 Hz, 2H), 2.73 (t, J = 5.9 Hz, 2H), 2.68 (d, J = 12.0 Hz, 1H), 2.64 (d, J = 4.7 Hz, 3H), 2.54 (d, J = 1.2 Hz, 3H), 2.43 – 2.35 (m, 2H), 2.33 – 2.27 (m, 2H), 1.96 (p, J = 6.3 Hz, 3H), 1.91 – 1.80 (m, 3H), 1.71 (d, J = 13.3 Hz, 3H), 1.56 – 1.43 (m, 4H), 1.35 (t, J = 6.8 Hz, 2H), 1.29 – 1.20 (m, 8H).

MNN-02-160:1- (1- (11- (1- (5-chloro-4- ((8-methoxy-1-methyl-3- (2- (methylamino) -2-oxoethoxy) -2-oxo-1, 2-dihydroquinolin-6-yl) amino) pyrimidin-2-yl) piperidine-4-carboxamido) undecyl) piperidin-4-yl) -3- (7- (difluoromethyl) -6- (1-methyl-1H-pyrazol-4-yl) -3, 4-dihydroquinolin-1 (2H) -yl) -N-methyl-1, 4,6, 7-tetrahydro-5H-pyrazolo [4,3-c ] pyridine-5-carboxamide. Synthesis of Compound MNN-02-160 using linker 11- ((tert-butoxycarbonyl) amino) undecanoic acid according to procedure A and subsequent passage through procedure B to give the title compound (0.89 mg, 7%, MS m/z 1221.82 [M+H]⁺).¹H NMR (500 MHz, DMSO-d⁶) δ = 9.07 (s, 1H), 8.10 (s, 1H), 7.96 (q, J = 4.8 Hz, 1H), 7.77 (t, J = 5.6 Hz, 1H), 7.74 (d, J = 0.9 Hz, 1H), 7.52 (s, 2H), 7.48 (d, J = 0.8 Hz, 1H), 7.08 (s, 1H), 7.01 (s, 1H), 6.79 (s, 1H), 6.76 (t, J = 55.3 Hz, 1H), 6.55 (s, 1H), 4.55 (s, 2H), 4.47 (m, J = 13.0 Hz, 3H), 4.31 (tt, J = 10.6, 4.5 Hz, 1H), 4.01 (s, 2H), 3.98 – 3.91 (m, 1H), 3.87 (s, 3H), 3.86 (s, 3H), 3.85 (s, 3H), 3.58 (m, J = 17.8, 5.3 Hz, 4H), 3.15 (t, J = 12.4 Hz, 1H), 2.99 (q, J = 6.6 Hz, 2H), 2.97 – 2.88 (m, 2H), 2.82 (t, J = 6.5 Hz, 2H), 2.73 (t, J = 6.0 Hz, 2H), 2.68 (d, J = 11.7 Hz, 1H), 2.64 (d, J = 4.6 Hz, 3H), 2.54 (d, J = 1.7 Hz, 3H), 2.42 – 2.34 (m, 1H), 2.33 – 2.27 (m, 2H), 1.96 (dt, J = 10.0, 6.3 Hz, 2H), 1.91 – 1.83 (m, 3H), 1.71 (d, J = 13.2 Hz, 3H), 1.49 (dq, J = 22.0, 8.1 Hz, 4H), 1.38 – 1.32 (m, 2H), 1.27 – 1.15 (m, 12H).

MNN-02-161:1- (1- (3- (2- (1- (5-chloro-4- ((8-methoxy-1-methyl-3- (2- (methylamino) -2-oxoethoxy) -2-oxo-1, 2-dihydroquinolin-6-yl) amino) pyrimidin-2-yl) piperidine-4-carboxamido) ethoxy) propionyl) piperidin-4-yl) -3- (7- (difluoromethyl) -6- (1-methyl-1H-pyrazol-4-yl) -3, 4-dihydroquinolin-1 (2H) -yl) -N-methyl-1, 4,6, 7-tetrahydro-5H-pyrazolo [4,3-c ] pyridine-5-carboxamide. Compound MNN-02-161 was synthesized according to procedure A using linker 3- (2- ((tert-butoxycarbonyl) amino) ethoxy) propionic acid and then passed through procedure B to give the title compound (2.52 mg, 20%, MS m/z 1152.48 [M+H]⁺).¹H NMR (500 MHz, DMSO-d⁶) δ = 8.83 (s, 1H), 8.05 (s, 1H), 7.96 (q, J = 4.6 Hz, 1H), 7.84 (t, J = 5.6 Hz, 1H), 7.73 (s, 1H), 7.53 (q, J = 2.3 Hz, 2H), 7.48 (s, 1H), 7.08 (s, 1H), 6.99 (s, 1H), 6.76 (t, 2H), 6.54 (d, J = 4.7 Hz, 1H), 4.54 (s, 2H), 4.47 (t, J = 18.1 Hz, 3H), 4.31 (d, J = 10.2 Hz, 1H), 3.98 (d, J = 18.1 Hz, 2H), 3.96 (s, 1H), 3.91 – 3.81 (m, 9H), 3.64 – 3.52 (m, 4H), 3.38 (t, J = 5.9 Hz, 2H), 3.17 (dt, J = 5.9, 2.8 Hz, 1H), 2.87 (s, 1H), 2.81 (d, 2H), 2.73 (t, J = 5.8 Hz, 2H), 2.72 – 2.68 (m, 1H), 2.64 (d, J = 4.7 Hz, 3H), 2.59 (dt, J = 12.3, 6.6 Hz, 1H), 2.54 (s, 3H), 2.44 – 2.36 (m, 2H), 1.95 (p, J = 6.3 Hz, 2H), 1.87 (d, J= 22.9 Hz, 3H), 1.69 (d, J = 9.7 Hz, 4H), 1.53 – 1.43 (m, 2H), 1.29 – 1.21 (m, 2H).

MNN-02-162:1- (1- (3- (2- (2- (1- (5-chloro-4- ((8-methoxy-1-methyl-3- (2- (methylamino) -2-oxoethoxy) -2-oxo-1, 2-dihydroquinolin-6-yl) amino) pyrimidin-2-yl) piperidine-4-carboxamido) ethoxy) propionyl) piperidin-4-yl) -3- (7- (difluoromethyl) -6- (1-methyl-1H-pyrazol-4-yl) -3, 4-dihydroquinolin-1 (2H) -yl) -N-methyl-1, 4,6, 7-tetrahydro-5H-pyrazolo [4,3-c ] pyridine-5-carboxamide. Synthesis of Compound MNN-02-162 using linker 2, 2-dimethyl-4-oxo-3,8,11-trioxa-5-aza-tetradecan-14-acid according to procedure A and subsequent passage through procedure B to give the title compound (2.91 mg, 20%, MS m/z 1196.40 [M+H]⁺).¹H NMR (500 MHz, DMSO-d⁶) δ = 8.85 (s, 1H), 8.06 (s, 1H), 7.95 (q, J = 4.6 Hz, 1H), 7.84 (t, J = 5.8 Hz, 1H), 7.73 (s, 1H), 7.53 (s, 2H), 7.48 (s, 1H), 7.08 (s, 1H), 7.00 (s, 1H), 6.77 (t, 1H), 6.53 (d, J = 5.1 Hz, 1H), 4.54 (s, 2H), 4.47 (dd, J = 26.3, 12.9 Hz, 3H), 4.35 – 4.27 (m, 1H), 4.00 (d, J = 5.1 Hz, 2H), 3.96 (d, J = 5.9 Hz, 1H), 3.90 – 3.82 (m, 9H), 3.60-3.56 (4H, attributed to HSQC ),3.48 (s, 2H), 3.37 (t, J = 5.9 Hz, 2H), 3.16 (q, J = 6.0 Hz, 3H), 2.88 (td, J = 13.0, 2.7 Hz, 2H), 2.82 (t, J = 6.4 Hz, 2H), 2.73 (q, J = 5.7 Hz, 2H), 2.69 (s, 1H), 2.64 (d, J = 4.7 Hz, 3H), 2.59 (q, J = 6.5 Hz, 2H), 2.54 (s, 3H), 2.40 (dh, J = 11.4, 3.7 Hz, 2H), 1.99 – 1.92 (m, 2H), 1.87 (d, J = 18.6 Hz, 3H), 1.77 – 1.66 (m, 3H), 1.48 (qd, J = 12.5, 4.1 Hz, 2H), 1.29 – 1.21 (m, 2H), 1.13 – 1.08 (m, 1H).

MNN-02-156 1- (1- (1- (5-chloro-4- ((8-methoxy-1-methyl-3- (2- (methylamino) -2-oxoethoxy) -2-oxo-1, 2-dihydroquinolin-6-yl) amino) pyrimidin-2-yl) piperidin-4-yl) -1-oxo-5, 8, 11-trioxa-2-aza-tetradec-yl) piperidin-4-yl) -3- (7- (difluoromethyl) -6- (1-methyl-1H-pyrazol-4-yl) -3, 4-dihydroquinolin-1 (2H) -yl) -N-methyl-1, 4,6, 7-tetrahydro-5H-pyrazolo [4,3-c ] pyridine-5-carboxamide. Synthesis of Compound MNN-02-156 using linker 2, 2-dimethyl-4-oxo-3,8,11,14-tetraoxa-5-azaheptadecane-17-acid according to procedure A and subsequent passage through procedure B to give the title compound .(2.00 mg, 20%, MS m/z 1241.04 [M+H]⁺).¹H NMR (500 MHz, DMSO-d⁶) δ = 8.83 (s, 1H), 8.06 (d, J = 1.4 Hz, 1H), 7.96 (d, J = 4.9 Hz, 1H), 7.85 (q, J = 6.2 Hz, 1H), 7.74 (d, J = 1.7 Hz, 1H), 7.53 (s, 2H), 7.48 (s, 1H), 7.08 (s, 1H), 7.00 (s, 1H), 6.91 – 6.63 (m, 2H), 6.53 (d, J = 4.7 Hz, 1H), 4.54 (s, 2H), 4.47 (dd, J = 27.5, 12.8 Hz, 3H), 4.31 (s, 1H), 4.00 (s, 2H), 3.97 (s, 1H), 3.89 – 3.83 (m, 9H), 3.61 (dt, J = 6.6, 3.4 Hz, 4H), 3.50 – 3.36 (m, 2H), 3.17 (p, J = 5.8 Hz, 3H), 2.88 (td, J= 12.8, 2.7 Hz, 1H), 2.82 (t, J = 6.1 Hz, 2H), 2.76 – 2.72 (m, 2H), 2.70 (d, J = 12.8 Hz, 1H), 2.65 (d, J = 4.7 Hz, 3H), 2.62 – 2.56 (m, 1H), 2.53 (d, J = 3.3 Hz, 3H), 2.45 – 2.33 (m, 1H), 2.30 (dd, J = 7.8, 5.2 Hz, 1H), 1.99 – 1.92 (m, 2H), 1.91 – 1.84 (m, 3H), 1.75 – 1.66 (m, 3H), 1.54 – 1.43 (m, 2H).

MNN-03-039:1- (1- (1 '- (1- (5-chloro-4- ((8-methoxy-1-methyl-3- (2- (methylamino) -2-oxoethoxy) -2-oxo-1, 2-dihydroquinolin-6-yl) amino) pyrimidin-2-yl) piperidine-4-carbonyl) - [1,4' -bipiperidine ] -4-carbonyl) piperidin-4-yl) -3- (7- (difluoromethyl) -6- (1-methyl-1H-pyrazol-4-yl) -3, 4-dihydroquinolin-1 (2H) -yl) -N-methyl-1, 4,6, 7-tetrahydro-5H-pyrazolo [4,3-c ] pyridine-5-carboxamide. Compound MNN-03-039 was synthesized according to procedure a using linker 1'- (tert-butoxycarbonyl) - [1,4' -bipiperidine ] -4-carboxylic acid and then passed through procedure B to give the title compound (0.88 mg, 10%, MS M/z 1232.68 [ m+h ] ⁺).

MNN-03-049:1- (1- (3- (1- (5-chloro-4- ((8-methoxy-1-methyl-3- (2- (methylamino) -2-oxoethoxy) -2-oxo-1, 2-dihydroquinolin-6-yl) amino) pyrimidin-2-yl) piperidine-4-carbonyl) -3-azaspiro [5.5] undecane-9-carbonyl) piperidin-4-yl) -3- (7- (difluoromethyl) -6- (1-methyl-1H-pyrazol-4-yl) -3, 4-dihydroquinolin-1 (2H) -yl) -N-methyl-1, 4,6, 7-tetrahydro-5H-pyrazolo [4,3-c ] pyridine-5-carboxamide. Synthesis of Compound MNN-03-049 using linker 3- (tert-butoxycarbonyl) -3-azaspiro [5.5] undecane-9-carboxylic acid according to procedure A and subsequent passage through procedure B to give the title compound .(2.65 mg, 30%, MS m/z 1216.57 [M+H]⁺).¹H NMR (500 MHz, DMSO-d⁶) δ = 9.11 (s, 1H), 8.11 (s, 1H), 7.94 (q, J = 4.6 Hz, 1H), 7.74 (s, 1H), 7.52 (d, J = 2.3 Hz, 1H), 7.51 – 7.45 (m, 2H), 7.09 (s, 1H), 7.01 (s, 1H), 6.81 (s, 1H), 6.77 (t, J = 55.2 Hz, 1H), 6.54 (s, 1H), 4.55 (s, 2H), 4.45 (d, J = 12.7 Hz, 3H), 4.40 – 4.28 (m, 1H), 4.02 (s, 3H), 3.89 – 3.84 (m, 9H), 3.71 – 3.58 (m, 4H), 3.56 (t, J = 5.6 Hz, 2H), 3.47 (s, 2H), 3.41 (s, 2H), 3.18 (t, J = 12.7 Hz, 1H), 3.00 (s, 2H), 2.93 (d, J = 10.5 Hz, 1H), 2.83 (t, J = 6.4 Hz, 2H), 2.74 (t, J = 5.7 Hz, 2H), 2.69 (s, 1H), 2.64 (d, J = 4.7 Hz, 3H), 2.60 (s, 1H), 2.54 (s, 3H), 1.96 (h, J = 6.8 Hz, 2H), 1.85 (d, J = 16.9 Hz, 2H), 1.73 – 1.63 (m, 4H), 1.50 (d, J = 16.4 Hz, 6H), 1.38 (s, 1H), 1.30 (s, 1H), 1.16 (dd, J = 24.9, 9.9 Hz, 3H).

MNN-03-050:1- (1- (2- (1- (5-chloro-4- ((8-methoxy-1-methyl-3- (2- (methylamino) -2-oxoethoxy) -2-oxo-1, 2-dihydroquinolin-6-yl) amino) pyrimidin-2-yl) piperidine-4-carbonyl) -2-azaspiro [3.3] heptane-6-carbonyl) piperidin-4-yl) -3- (7- (difluoromethyl) -6- (1-methyl-1H-pyrazol-4-yl) -3, 4-dihydroquinolin-1 (2H) -yl) -N-methyl-1, 4,6, 7-tetrahydro-5H-pyrazolo [4,3-c ] pyridine-5-carboxamide. Synthesis of Compound MNN-03-050 using linker 2- (tert-butoxycarbonyl) -2-azaspiro [3.3] heptane-6-carboxylic acid according to procedure 1 and subsequent passage through procedure 2 to give the title compound (4.72 mg, 40%, MS m/z 1160.73 [M+H]⁺).¹H NMR (500 MHz, DMSO-d⁶) δ = 9.17 (s, 1H), 8.12 (s, 1H), 7.94 (q, J = 4.7 Hz, 1H), 7.74 (s, 1H), 7.52 (dd, J = 7.1, 2.3 Hz, 1H), 7.48 (d, J = 2.6 Hz, 2H), 7.09 (s, 1H), 7.00 (s, 1H), 6.81 (d, J = 2.2 Hz, 1H), 6.77 (t, 1H), 6.54 (s, 1H), 4.55 (s, 2H), 4.42 (d, J = 12.8 Hz, 3H), 4.32 (dq, J = 11.2, 5.3 Hz, 1H), 4.24 (s, 1H), 4.22 (s, 1H), 4.08 (s, 1H), 4.06 (s, 2H), 4.02 (s, 2H), 3.86 (dd, J = 4.5, 1.7 Hz, 9H), 3.80 (d, J = 13.9 Hz, 1H), 3.70 (s, 1H), 3.58 (dt, J = 18.3, 6.3 Hz, 4H), 3.26 (td, J = 8.4, 2.7 Hz, 1H), 3.11 (s, 1H), 2.97 (s, 2H), 2.83 (t, J = 6.4 Hz, 2H), 2.73 (h, J = 6.2 Hz, 3H), 2.64 (d, J = 4.7 Hz, 3H), 2.54 (s, 3H), 2.34 (td, J = 8.1, 4.0 Hz, 2H), 2.25 (d, J = 8.2 Hz, 1H), 1.97 (p, J = 6.1 Hz, 2H), 1.87 (d, J = 13.2 Hz, 2H), 1.81 (s, 1H), 1.69 (d, J = 31.1 Hz, 3H), 1.44 (t, J = 12.0 Hz, 2H).

MNN-03-038:1- (1- (3- (4- (1- (5-chloro-4- ((8-methoxy-1-methyl-3- (2- (methylamino) -2-oxoethoxy) -2-oxo-1, 2-dihydroquinolin-6-yl) amino) pyrimidin-2-yl) piperidine-4-carbonyl) piperazin-1-yl) propionyl) piperidin-4-yl) -3- (7- (difluoromethyl) -6- (1-methyl-1H-pyrazol-4-yl) -3, 4-dihydroquinolin-1 (2H) -yl) -N-methyl-1, 4,6, 7-tetrahydro-5H-pyrazolo [4,3-c ] pyridine-5-carboxamide. Compound MNN-03-038 was synthesized according to procedure 1 using linker 3- (4- (tert-butoxycarbonyl) piperazin-1-yl) propionic acid and then passed through procedure 2 to give the title compound (3.33 mg, 30% yield , MS m/z 1177.74 [M+H]⁺).¹H NMR (500 MHz, DMSO-d⁶) δ = 9.57 (s, 1H), 8.94 (s, 1H), 8.09 (s, 1H), 7.98 (d, J = 4.7 Hz, 1H), 7.74 (s, 1H), 7.58 (d, J = 2.2 Hz, 1H), 7.50 (d, J = 2.3 Hz, 1H), 7.49 (d, J = 0.8 Hz, 1H), 7.09 (s, 1H), 6.99 (s, 1H), 6.77 (t, J = 55.2 Hz, 1H), 6.76 (s, 1H), 6.55 (s, 1H), 4.55 (s, 2H), 4.48 (m, J = 15.8 Hz, 4H), 4.41 – 4.32 (m, 1H), 4.23 (s, 1H), 4.02 (s, 2H), 3.95 (d, J = 13.1 Hz, 1H), 3.87 (s, 3H), 3.86 (s, 6H), 3.80-3.50 (7H, as attributed to HSQC ), 3.36 (s, 2H), 3.26 – 3.17 (m, 1H), 3.10 (s, 1H), 3.04 – 2.94 (m, 5H), 2.91 (q, J = 6.8 Hz, 2H), 2.83 (q, J = 8.5 Hz, 3H), 2.76 (q, J = 8.1 Hz, 2H), 2.64 (d, J = 4.7 Hz, 3H), 2.54 (s, 3H), 1.94 (m, J= 13.2, 4.8 Hz, 5H), 1.84 – 1.63 (m, 3H), 1.50 (s, 2H).

MNN-03-041- (1- (4- (1- (5-chloro-4- ((8-methoxy-1-methyl-3- (2- (methylamino) -2-oxoethoxy) -2-oxo-1, 2-dihydroquinolin-6-yl) amino) pyrimidin-2-yl) piperidine-4-carboxamido) cyclohexane-1-carbonyl) piperidin-4-yl) -3- (7- (difluoromethyl) -6- (1-methyl-1H-pyrazol-4-yl) -3, 4-dihydroquinolin-1 (2H) -yl) -N-methyl-1, 4,6, 7-tetrahydro-5H-pyrazolo [4,3-c ] pyridine-5-carboxamide. Compound MNN-03-041 was synthesized according to procedure 1 using linker 4- ((tert-butoxycarbonyl) amino) cyclohexane-1-carboxylic acid and then passed through procedure 2 to give the title compound (4.73 mg, 40%, MS m/z 1162.38 [M+H]⁺).¹H NMR (500 MHz, DMSO-d⁶) δ = 8.96 (s, 1H), 8.09 (s, 1H), 7.94 (q, J = 4.8 Hz, 1H), 7.77 – 7.71 (m, 1H), 7.55 – 7.49 (m, 2H), 7.48 (t, J = 1.4 Hz, 1H), 7.08 (s, 1H), 7.04 – 6.98 (m, 1H), 6.81 (s, 1H), 6.76 (t, J = 55.3 Hz, 1H), 6.54 (s, 1H), 4.55 (d, J = 2.3 Hz, 2H), 4.50 (d, J = 13.2 Hz, 3H), 4.32 (dq, J = 10.8, 5.4 Hz, 1H), 4.02 (s, 3H), 3.90 – 3.80 (m, 9H), 3.77 (s, 1H), 3.60 (t, J = 5.7 Hz, 2H), 3.56 (t, J = 5.6 Hz, 2H), 3.43 (s, 1H), 3.18 (t, J = 12.7 Hz, 1H), 3.07 (t, J = 11.4 Hz, 1H), 2.98 (t, J = 12.4 Hz, 1H), 2.92 – 2.85 (m, 2H), 2.85 – 2.79 (m, 2H), 2.74 (t, J = 6.0 Hz, 2H), 2.69 (d, J = 3.3 Hz, 1H), 2.67 – 2.59 (m, 3H), 2.54 (s, 3H), 1.96 (q, J = 5.9 Hz, 2H), 1.88 (s, 3H), 1.68 (d, J = 12.7 Hz, 6H), 1.48 (d, J= 29.4 Hz, 4H).

MNN-03-040:1- (1- (1- (1- (5-chloro-4- ((8-methoxy-1-methyl-3- (2- (methylamino) -2-oxoethoxy) -2-oxo-1, 2-dihydroquinolin-6-yl) amino) pyrimidin-2-yl) piperidine-4-carbonyl) azetidine-3-carbonyl) piperidin-4-yl) -3- (7- (difluoromethyl) -6- (1-methyl-1H-pyrazol-4-yl) -3, 4-dihydroquinolin-1 (2H) -yl) -N-methyl-1, 4,6, 7-tetrahydro-5H-pyrazolo [4,3-c ] pyridine-5-carboxamide. Compound MNN-03-040 was synthesized according to procedure 1 using linker 1- (tert-butoxycarbonyl) azetidine-3-carboxylic acid and then passed through procedure 2 to give the title compound (0.82 mg, 6%, MS m/z 1120.61 [M+H]⁺).¹H NMR (500 MHz, DMSO-d⁶) δ = 8.89 (s, 1H), 8.07 (d, J = 2.2 Hz, 1H), 7.97 – 7.92 (m, 1H), 7.74 (d, J = 8.5 Hz, 1H), 7.54 (t, J = 2.8 Hz, 1H), 7.51 (d, J = 2.2 Hz, 1H), 7.48 (d, J = 4.7 Hz, 1H), 7.08 (s, 1H), 6.98 (d, J = 5.2 Hz, 1H), 6.79 (d, J = 3.1 Hz, 1H), 6.78 (td, J= 55.2, 3.1 Hz, 1H), 6.54 (s, 1H), 4.55 (d, J = 3.4 Hz, 2H), 4.48 (d, J = 13.8 Hz, 3H), 4.31 (dt, J = 14.3, 6.9 Hz, 1H), 4.01 (d, J = 5.2 Hz, 2H), 3.99 (s, 1H), 3.94 – 3.88 (m, 1H), 3.88 – 3.81 (m, 9H), 3.74 (ddd, J = 15.0, 8.5, 6.0 Hz, 1H), 3.67 (d, J = 13.5 Hz, 1H), 3.60 (s, 2H), 3.56 (t, J = 5.7 Hz, 2H), 3.21 – 3.12 (m, 2H), 3.09 (td, J = 7.3, 4.8 Hz, 2H), 2.94 (t, J = 13.0 Hz, 2H), 2.83 (t, J = 6.5 Hz, 2H), 2.79 (s, 1H), 2.74 (t, J = 5.9 Hz, 2H), 2.64 (dd, J = 4.7, 2.2 Hz, 3H), 2.58 – 2.52 (m, 3H), 1.96 (t, J = 5.8 Hz, 2H), 1.90 (s, 3H), 1.76 (d, J = 7.3 Hz, 1H), 1.66 (t, J = 14.3 Hz, 2H), 1.43 (d, J = 13.7 Hz, 2H).

MNN-04-022 (S) -1- (1- (3- (4- (1- (5-chloro-4- ((2-cyclopropyl-3, 3-difluoro-7-methyl-6-oxo-1, 2,3,4,6, 7-hexahydro- [1,4] oxazepino [2,3-c ] quinolin-10-yl) amino) pyrimidin-2-yl) piperidine-4-carbonyl) piperazin-1-yl) propionyl) piperidin-4-yl) -3- (7- (difluoromethyl) -6- (1-methyl-1H-pyrazol-4-yl) -3, 4-dihydroquinolin-1 (2H) -yl) -N-methyl-1, 4,6, 7-tetrahydro-5H-pyrazolo [4,3-c ] pyridine-5-carboxamide. Compound MNN-04-022 was synthesized according to procedure 1 using linker 3- (4- (tert-butoxycarbonyl) piperazin-1-yl) propionic acid to give 3- (7- (difluoromethyl) -6- (1-methyl-1H-pyrazol-4-yl) -3, 4-dihydroquinolin-1 (2H) -yl) -N-methyl-1- (1- (3- (piperazin-1-yl) propionyl) piperidin-4-yl) -1,4,6, 7-tetrahydro-5H-pyrazolo [4,3-c ] pyridine-5-carboxamide (9.90 mg, 97%) as a white solid.

To a solution of (S) -1- (5-chloro-4- ((2-cyclopropyl-3, 3-difluoro-7-methyl-6-oxo-1, 2,3,4,6, 7-hexahydro- [1,4] oxazepino [2,3-C ] quinolin-10-yl) amino) pyrimidin-2-yl) piperidine-4-carboxylic acid (11.1 mg, 0.0199 mmol, 1.20 eq.) in DMF (330 μl, 0.05M) was added DIPEA (20.2 μl, 0.0116 mmol, 7.00 eq.) and HATU (8.18 mg, 0.0215 mmol, 1.30 eq.) and the mixture was stirred under nitrogen at 0 ℃ for 15 min. After stirring for 15 min, the crude mixture of 3- (7- (difluoromethyl) -6- (1-methyl-1H-pyrazol-4-yl) -3, 4-dihydroquinolin-1 (2H) -yl) -N-methyl-1- (1- (3- (piperazin-1-yl) propionyl) piperidin-4-yl) -1,4,6, 7-tetrahydro-5H-pyrazolo [4,3-c ] pyridine-5-carboxamide dissolved in DMF was added dropwise and the reaction stirred for 1H to give (S) -1- (1- (3- (4- (5-chloro-4- ((2-cyclopropyl-3, 3-difluoro-7-methyl-6-oxo-1, 2,3,4,6, 7-hexahydro- [1,4] oxazao [2,3-c ] quinolin-10-yl) amino) piperidin-4-carbonyl) piperidin-4-piperazin-1-yl) propionyl) piperidin-4-yl) -3- (difluoro-4- ((2-cyclopropyl-3, 3-difluoro-7-methyl-6-oxo-1, 2, 3-hexahydro- [1,4] oxazino [2, 3-amino) pyrimidin-4-carbonyl) piperidin-4-yl ] piperidin-3- (1- (5-methyl) -5-yl) -pyrazolo-1- (1, 3-dimethyl) -amino-5-methyl-5-butan-amide, 9%, MS M/z 1207.86 [ M+H ] ⁺).

Example 2 reporter Gene activation

The compounds shown above were tested for their ability to activate the BCL6 reporter GFP integrated into the KARPAS lymphoma cell line. The reporter construct contains 8 high confidence BCL6 binding sites and flanking base pairs identified by chromatin immunoprecipitation sequencing (ChIP-seq) and functional genomic data at a validated cell death gene upstream of the minimal promoter driving Green Fluorescent Protein (GFP) as described in Gourisankar et al Nature 620, 417-425 (2023). The reporter gene is integrated into the genome of other genetically unmodified KARPAS422 DLBCL cells and GFP is not normally transcribed due to the repressing function of endogenous BCL 6. As shown in fig. 4 and 8C, several compounds activated the reporter gene even at the lowest concentration.

EXAMPLE 3 cell viability assay

After 72 hours of treatment, HAT-BCL6 TCIP was tested for its ability to kill lymphoma cells. 30,000 cells were plated in 100 μl of medium per well of a 96-well plate and treated with the drug for the indicated times and doses. The fluorescence ratio at 560/590nm was recorded 1.5 hours after addition of the resazurin-based cell indicator. After subtraction of background fluorescence, signals were normalized to DMSO-treated cells. DMSO content was normalized to 0.1%. EC50 values were calculated using standard 4-parameter logarithmic logic functions. The data are shown in fig. 5A and 8A (sudfl 5 cells) and fig. 5B (KARPAS 422 cells), with EC50 values in fig. 8A and 8B and table 1. dCBP-1 is a degradation agent for p300/CBP (CAS number 2484739-25-3), A485 is a catalytic site p300/CBP inhibitor (CAS number 1889279-16-6), and GNE-781 is a p300/CBP bromodomain inhibitor (CAS number 1936422-33-1). Negative 1 is a negative control for MNN-02-155 in which BCL6 binding is eliminated, the compound has the structure:

。

TABLE 1 EC ₅₀ values in KARPAS422 cells

EXAMPLE 4 ternary complex formation

To determine whether HAT-BCL6 TCIP works by inducing a ternary complex between p300/CBP and BCL6, each compound was tested for its ability to rescue loss of cell viability generated with one MNN-03-038 (TCIP 3) of the most potent HAT-BCL6 TCIP against the excess p300/CBP inhibitor (including SGC-CBP-30, GNE-781 or a-485) added at the beginning of the experiment and against the excess BCL6 inhibitor (including BI-3812, GSK137 (CAS 2728727-91-9) or CCT nM 67 (CAS 2378853-67-7)). The data are shown in fig. 6A (KARPAS 422 cells) and fig. 6B and fig. 8E (SUDHL 5 cells). Ternary complex formation reactions are on the rescue of viability because excess monomeric drugs compete for bioactive ternary complexes.

Example 5 half-life and internal clearance

Table 3 shows the in vitro half-life and internal clearance of p300/CBP TCIP in a mouse liver microsome stability assay relative to the reference kinase inhibitor sunitinib.

Table 3.

EXAMPLE 6 oncogene suppression Studies

Method of

And (5) culturing the cells. Lymphoma and leukemia cells were cultured in RPMI-1640 (ATCC 30-2001) +10% FBS with antibiotic (100 XPenStrep GIBCO # 15140122) and incubated at 37℃and 5% CO ₂. Cells were routinely checked for mycoplasma and immediately checked once suspected. No cultures tested positive.

Western blotting. Cells were plated at 100 ten thousand per milliliter (mL) and treated with drugs at the indicated time points and doses. Cells were harvested on ice in RIPA buffer (50 mM Tris-HCl pH 8, 150mM NaCl, 1% NP-40, 0.1% DOC, 1% SDS, protease inhibitor cocktail (about 20 mg/ml pepstatin, aprotinin and leupeptin), 1mM DTT) and 1:200 benzonase (Sigma #E1014) was added and incubated 20 min. After centrifugation at 14,000g and 4 ℃ for 10 min, the supernatant was collected and the protein concentration was measured by Bradford. Antibodies used for immunoblotting were ：BCL6 (Cell Signaling D65C10)、p300 (Santa Cruz F-4)、CBP (Cell Signaling D6C5)、c-MYC (Cell Signaling D84C12) and GAPDH (Santa Cruz 6C 5). All antibodies were used at a dilution of 1:1000 v/v except GAPDH (1:2000). Blotting imaging and quantification was performed using ImageStudio (Licor).

RNA extraction, qPCR and sequencing library preparation. Cells were plated at 100 ten thousand/ml and harvested in TRIsure (Bioline # 38033). RNA was extracted using a Direct-zol RNA MicroPrep column (zymo#R2062) treated with DNase I. cDNA for RT-qPCR was prepared using SENSIFAST CDNA preparation kit according to the manufacturer's instructions (Bioline # 65054). mu.L of cDNA was used for each RT-qPCR reaction and prepared with SYBR Lo-ROX (Bioline # 94020). For sequencing library preparation, transcripts containing polyA (neb#e7490S) were enriched, prepared as a double-ended library (neb#e7760S), and sequenced from Novogene on Illumina NovaSeq using double-ended 150 bp reads.

RNA-seq analysis. The quality of the original reading was checked using fastqc (www.bioinformatics.babraham.ac.uk/projects/fastqc /), and trimmed from the adapter using parameters cutadapt -a AGATCGGAAGAGCACACGTCTGAACTCCAGTCA (SEQ ID NO: 1) -b AGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGT (SEQ ID NO: 2) --nextseq- using cutadapt (Martin, embnet. Journ 17, 10 (2011)) for trim = 20—minimum-length 1. Transcripts were quantified for human Gencode v33 index transcriptome and annotation using kallisto (brain et al Nature Biotechnology, 525-527 (2016)). Differential gene analysis was performed using DESeq2 (Love et al Genome Biol 15, 550 (2014)) using apeglm (Zhu et al bioenformatics 35, 2084-2092 (2019)) to reduce log ₂ fold changes.

Results

The results are shown in fig. 7A to 7G. The data in fig. 7A-7F show a reduction in c-MYC compared to control (such as BCL6 inhibition by BI-3812 (nanomolar inhibitor of BCL 6) (or related compounds), or p300/CBP bromodomain inhibition by GNE-781 (nanomolar inhibitor of p 300/CBP) (or related compounds)). The data in fig. 7E shows that ¬ c-MYC is the most significantly suppressed gene at 1 hour of 1 nM treatment in an unbiased analysis of all transcripts altered by TCIP 3. The data in fig. 7G shows that this decrease is dependent on BCL6 levels of the cell line. The data in fig. 7A show that the containment of MYC does not occur through degradation or loss of the target protein (p 300, CBP, BCL 6), but through a new signaling event. The data in FIG. 7D shows that suppression of c-MYC is first at the transcriptional level of the c-MYC gene, as expected for TCIP acting by modulating transcription.

Example 7 ternary Complex dependent, BCL6 regulated intracellular Gene expression and cytotoxicity assays

Method of

Cell viability measurement. 30,000 cells in 100 μl of medium were seeded in 96-well plates and treated with the indicated doses of drug for 72 hours. A resazurin-based cell health indicator (PrestoBlue, thermoFisher #p50200) was added, at which time the plates were incubated at 37 ℃ for 1.5 hours. The fluorescence ratio at 560/590nm was then recorded. After subtraction of background fluorescence, signals were normalized to DMSO-treated cells. The dose response curve fitting and statistical analysis of the data was performed using a four parameter logarithmic logic function using the drc package in R.

And (5) measuring a reporter gene. KARPAS422 cells were lentivirally transduced with the previously reported GFP reporter construct (Gourisankar et al Nature 620, 417-425 (2023)). After selection, cells were plated and treated with the indicated amounts of each compound for 8 hours. Cells were washed in 2.5% FBS/PBS, 7-AAD at 1:250 v/v was administered to differentiate between live and dead cells, and harvested for flow cytometry on BD Accuri. In view of the transduced polyclonal population, the area under the histogram curve representing FITC signals of all living cells was calculated as a comprehensive measure of total GFP signal. GFP-positive gates were drawn from untransduced cells and the area of each sample exceeding the threshold was calculated and normalized to DMSO-treated cells.

TR-FRET. A mixture of 10 nM p 300-bromodomain (purified from Addgene construct # 39018, available from Nicola Burgess-Brown), 200 nM biotinylated BCL6-BTB, 20 nM streptavidin-FITC (Thermo #SA 1001) and 1:400 anti-6 xHis terbium antibody (Perkinelmer #61HI2 TLF) in 10. Mu.L buffer (20 mM HEPES, 150 mM NaCl, 0.1% BSA, 0.1% NP-40 and 1mM TCIP) was added to a low-volume 384-well plate. Each protein mixture was incubated with digitally dispensed drug (Tecan D300 e) for 1 hour at room temperature in the dark, then challenged at 337 nM and the emissions at 520 nM (FITC) and 490 nM (terbium) were measured with a PHERASTAR FS plate reader (BMG Labtech). The signal ratios at 520 nm and 490 nm were calculated and normalized to DMSO treatment conditions and plotted.

Results

The results are shown in figures 8A to 8F and show that MNN-03-038 (TCIP 3) exhibits greater cell killing than the inhibitor alone and that cell killing is dependent on ternary complex formation between p300 and BCL 6. FIG. 8A shows the enhanced TCIP3 induced cell killing compared to known p300/CBP inhibitors (A-485), degradation agents (dCBP-1) or combinations of BCL6 and p300/CBP inhibitors (BI-3812 and GNE-781, respectively). FIG. 8B shows IC ₅₀ values and structure activity relationship of all p300/CBP TCIP molecules synthesized in a 72 hour viability assay in SUDHL5 cells. FIG. 8C shows that treatment with TCIP3 induces transcriptional activation of the BCL6 promoter relative to BI-3812 or GNE-781. FIG. 8D shows that all TCIPs are capable of biochemically inducing a ternary complex between p300 and BCL6 in the TR-FRET assay, relative to BI-3812 and GNE-781, both for linear and rigid linkers. FIG. 8E shows that titrating BI-3812 or GNE-781 in a dose-dependent manner can rescue cell killing by TCIP3 in a 72 hour viability assay. Fig. 8F shows TCIP3 killing was more effective in DLBCL cell lines with high levels of BCL6 protein.

All references, including publications, patent applications, and patents, cited herein are hereby incorporated by reference to the same extent as if each reference were individually and specifically indicated to be incorporated by reference and were set forth in its entirety herein.

Claims

1. A compound of formula (I)

A-L-B (I)

Or a pharmaceutically acceptable salt thereof, wherein:

a is a BCL6 binding moiety;

L is a linker, and

B is the moiety that binds to the bromodomain of histone acetyltransferase.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein a is a moiety of the formula:

、、

、、、

、 Or (b) 。

3. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein a is a moiety of the formula:

。

4. the compound of any one of claims 1-3, or a pharmaceutically acceptable salt thereof, wherein B is a moiety that binds to the bromodomain of p300 or CBP.

5. The compound of any one of claims 1-4, or a pharmaceutically acceptable salt thereof, wherein B is a moiety of the formula:

、、、

、 Or (b) 。

6. The compound of any one of claims 1-5, or a pharmaceutically acceptable salt thereof, wherein B is a moiety of the formula:

。

7. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is a compound of formula (Ia):

(Ia)。

8. the compound of any one of claims 1-7, or a pharmaceutically acceptable salt thereof, wherein L is a direct bond, or comprises any combination of-CH ₂-、-CH=CH-、-C≡C-、-O-、-NR'-、-BR'-、-S-、-C(O)-、-C(NR')-、-S(O)-、-S(O)₂ -, arylene, heteroarylene, cycloalkylene, and heterocyclylene moieties, wherein the arylene, heteroarylene, cycloalkylene, and heterocyclylene moieties are independently unsubstituted or substituted with 1, 2, or 3 substituents.

9. The compound of any one of claims 1-8, or a pharmaceutically acceptable salt thereof, wherein L is a direct bond, or comprises any combination of-CH ₂-、-O-、-C(O)-、-NH-、-N(CH₃ -, cycloalkylene-, and heterocyclylene moieties.

10. The compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, wherein L comprises any combination of the following moieties:

、-(CH₂)_q-、-C(O)-、-NH-、、、、、、,

11. The compound of any one of claims 1-10, or a pharmaceutically acceptable salt thereof, wherein L has a formula selected from the group consisting of:

、 And ;

12. The compound of any one of claims 1-10, or a pharmaceutically acceptable salt thereof, wherein L has a formula selected from the group consisting of:

And 。

13. The compound of claim 1, wherein the compound is selected from the group consisting of:

and pharmaceutically acceptable salts thereof.

14. A pharmaceutical composition comprising a compound of any one of claims 1-13, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

15. A method of treating a proliferative disease in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-13, or a pharmaceutically acceptable salt thereof.

16. The method of claim 14, wherein the proliferative disease is a cancer selected from the group consisting of carcinoma, sarcoma, and hematological malignancy.

17. The method of claim 15, wherein the cancer is lymphoma.

18. The method of claim 16, wherein the cancer is diffuse large B-cell lymphoma.

19. A compound of any one of claims 1-13, or a pharmaceutically acceptable salt thereof, for use as a medicament.

20. A compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, for use in the treatment of a proliferative disease.

21. The compound of claim 20, wherein the proliferative disease is a cancer selected from the group consisting of carcinoma, sarcoma, and hematological malignancy.

22. The compound of claim 21, wherein the cancer is lymphoma.

23. The compound of claim 22, wherein the cancer is diffuse large B-cell lymphoma.

24. A kit comprising a compound of any one of claims 1-13, or a pharmaceutically acceptable salt thereof.