WO2025160079A1

WO2025160079A1 - Indazolo-amino-pyrimidinone compounds

Info

Publication number: WO2025160079A1
Application number: PCT/US2025/012443
Authority: WO
Inventors: Magnus Munck AF ROSENSCHÖLD; Filip Alexander PAULSEN; Andrea ÖHLANDER; John Henry Van Duzer; Joon O. Jung; Lei Shi; Peter Blume-Jensen; Åsa Helén NILSSON; Caroline Maria WIGERUP; David Proia; Kristina MASSON
Original assignee: Acrivon Therapeutics Inc
Current assignee: Acrivon Therapeutics Inc
Priority date: 2024-01-22
Filing date: 2025-01-21
Publication date: 2025-07-31
Anticipated expiration: 2026-07-22

Abstract

Indazolo-amino-pyrimidinone compounds of formula I; (I), for use in methods of inhibiting both Wee1 A kinase and Myt1 kinase.

Description

INDAZOLO-AMINO-PYRIMIDINONE COMPOUNDS

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to United States Provisional Application No. 63/623,648, filed January 22, 2024, the entirety of which is incorporated herein by reference.

BACKGROUND

[0002] Cancer cells frequently have a defective Gl/S checkpoint, often via disrupted p53 activity due to mutations or deletion, or inactivation by viral oncoproteins. Therefore, cancer cells rely heavily on other cell cycle checkpoints, including the G2/M checkpoint, to avoid accumulation of deleterious DNA damage and mitotic catastrophe. As such, cancer cells are hypothesized to be particularly vulnerable to inhibition of proteins that safeguard the entry into mitosis. Matheson, C. J. et al Trends Pharmacol Sci 37, 872-881 (2016).

[0003] Inhibition of Weel A kinase has been explored as a therapeutic option for the treatment of cancers. Weel A kinase is a tyrosine kinase belonging to the Weel kinase family, including Weel A kinase, WeelB kinase, and Mytl kinase. Rora, A. G. L. et al J Hematol Oncol 13, 126 (2020). The primary role for this kinase family is to regulate cell cycle progression and entry into mitosis (Weel A kinase and Mytl kinase) or meiosis (WeelB kinase). The key complex regulating mitotic entry is Cdkl/cyclin Bl complex, also known as the mitosis-promoting factor. WeelA kinase constrains Cdkl/cyclin Bl complex activity by phosphorylating Cdkl on the inhibitory tyrosine 15 site (Y¹⁵). Hence, inhibition of WeelA kinase effectively promotes Cdkl/cyclin Bl complex activity by preventing inhibitory Y¹⁵ phosphorylation. Untimely activation of Cdkl/cyclin B complex promotes premature entry into mitosis with unresolved DNA damages, ultimately leading to mitotic catastrophe and cell death.

[0004] Several WeelA kinase inhibitors are currently being tested in clinical trials (Bukhari, A. B. et al Frontiers Oncol 12, 828684 (2022)) and have shown activity in many indications. A phase II study of the WeelA kinase inhibitor AZD1775 (adavosertib) has shown promising results in women with uterine serous carcinoma. Liu, J. F. et al J Clin Oncol 39, 1531-1539 (2021). Adavosertib has also shown effect compared to active monitoring in RAS/TP53 mutated metastatic colorectal cancer. Seligmann, J. F. et alJC Oncol 39, 3705-3715 (2021). In a phase lb trial of 18 patients with platinum-resistant ovarian cancer, the combination of ZN-c3 (azenosertib) plus paclitaxel led to an ORR of 50% (J Clin Oncol 41, 2023 (suppl 16; abstr 5513)). Despite these results, Weel A kinase inhibitors have been plagued by adverse effects particularly when used in combination with chemotherapy (Kong A., Mehanna H. Curr. Oncol. Rep. 2021; 23: 107 and Martorana F., et al. Cancers (Basel). 2022; 14:953).

[0005] More recently it has been shown that a combination of low doses of a Weel A kinase inhibitor and a PMytl inhibitor exhibited synergistic effects in promoting CDK activation, exacerbating replication stress and replication catastrophe and in eradicating ovarian cancer cells and organoid models. (Benada J, et al. NAR Cancer. 2023 Jun 13;5(3):zcad029. doi: 10.1093/narcan/zcad029. PMID: 37325550; PMCID: PMC10262308). While WeelA kinase phosphorylates both Cdkl and Cdk2 on ty rosine 15 (Tyrl5), PMytl only phosphorylates cyclin-dependent kinase 1 (Cdkl), primarily at threonine 14 (Thrl4) and to some extent at Tyrl5. Phosphorylation of these sites inhibits Cdkl ’s kinase activity and its ability, in complex with cyclin Bl, to trigger mitosis. Thus, the main biological function of Mytl kinase is to prevent replication of cells with high levels of damaged DNA or perturbed checkpoints. Cancers with amplification of CCNE1, a cyclin that drives entry and progression of S phase, have been shown to be highly sensitive to Mytl kinase inhibition (Gallo, D. et al. CCNE1 amplification is synthetic lethal with PKMYT1 kinase inhibition. Nature 604, 749-756 (2022)). Similarly, mutations in FBXW7 that deactivate the ubiquitin ligase responsible for cyclin El degradation, may also result in abnormally high cyclin El levels.

[0006] Given the encouraging signs of the synergies resulting from simultaneous Weel A kinase and Mytl kinase inhibition, we sought to develop compounds that exhibit both inhibitory' activities.

SUMMARY

[0007] In some embodiments, the present disclosure provides a compound having structural formula I: solvate, enantiomer, tautomer, or diastereomer thereof, or a pharmaceutically acceptable salt of any of the foregoing wherein, wherein each of X, Y, Z, R¹, and R² are as defined below and described herein.

[0008] In some embodiments, the present disclosure provides a pharmaceutical composition comprising a compound of formula I, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0009] In some embodiments, the present disclosure provides a method of simultaneously inhibiting both WeelA kinase and Mytl kinase in a patient or in a biological sample, the method comprising administering to the patient or contacting the biological sample with a compound of formula I, or a pharmaceutically acceptable salt thereof.

[0010] In some embodiments, the present disclosure provides a method of treating a disease or disorder associated with both WeelA kinase and Mytl kinase, the method comprising administering to a patient in need thereof a compound of formula I, or a pharmaceutically acceptable salt thereof.

[0011] In some such embodiments, the disease or disorder associated with WeelA kinase and Mytl kinase is a cancer. In some embodiments, the cancer is selected from a brain cancer, a cervicocerebral cancer, a cardiac cancer, a gastrointestinal cancer, an esophageal cancer, a thyroid cancer, a small cell cancer, a non-small cell cancer, a breast cancer, a lung cancer, a stomach cancer, a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms' cancer, a skin cancer, malignant melanoma, a neuroblastoma, an osteosarcoma, an Ewing's tumor, a soft part sarcoma, an acute leukemia, a chronic lymphatic leukemia, a chronic myelocytic leukemia, polycythemia vera, a malignant lymphoma, multiple myeloma, a Hodgkin's lymphoma, and a non- Hodgkin's lymphoma.

DETAILED DESCRIPTION

1. Definitions

[0012] As used herein, the following definitions shall apply unless otherwise indicated. For purposes of this disclosure, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75^th Ed. Additionally, general principles of organic chemistry are described in "‘Organic Chemistry⁷’. Thomas Sorrell, University Science Books, Sausalito: 1999, and “March’s Advanced Organic Chemistry'’, 5^th Ed., Ed.: Smith, M.B. and March, J., John Wiley & Sons, New York: 2001, the entire contents of which are hereby incorporated by reference.

[0013] The term “aliphatic” or “aliphatic group”, as used herein, means a straight-chain (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is completely saturated or that contains one or more units of unsaturation, or a monocyclic hydrocarbon or bicyclic hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic (also referred to herein as “carbocyclyl” “cycloaliphatic” or “cycloalkyl”), that has a single point of attachment to the rest of the molecule. Unless otherwise specified, aliphatic groups contain 1-6 aliphatic carbon atoms. In some embodiments, aliphatic groups contain 1-5 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-4 aliphatic carbon atoms. In still other embodiments, aliphatic groups contain 1-3 aliphatic carbon atoms, and in yet other embodiments, aliphatic groups contain 1-2 aliphatic carbon atoms. In some embodiments, “cycloaliphatic” (or “carbocyclyl” or “cycloalkyl”) refers to a monocyclic C3-C6 hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic, that has a single point of attachment to the rest of the molecule. Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups and hybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl.

[0014] The term “alkyl”, as used herein, means a straight or branched hydrocarbon chain that is completely saturated. Exemplary alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, etc.

[0015] The term “heteroatom” means one or more of oxygen, sulfur, nitrogen, phosphorus, or silicon (including, any oxidized form of nitrogen, sulfur, phosphorus, or silicon; the quatemized form of any basic nitrogen or; a substitutable nitrogen of a heterocyclic ring, for example N (as in 3.4-dihydro-2H-pyrrolyl). NH (as in pyrrohdinyl) or NR⁺ (as inN-substituted pyrrolidinyl)).

[0016] The term “unsaturated”, as used herein, means that a moiety has one or more units of unsaturation.

[0017] As used herein, the term “partially unsaturated” refers to a moiety that includes at least one double or triple bond. The term “partially unsaturated” is intended to encompass moieties having multiple sites of unsaturation, but when used to describe a ring, is not intended to include aryl or heteroaryl moieties, as herein defined.

[0018] The term “saturated” refers to a moiety that has no double or triple bonds. [0019] The term “alkylene’' refers to a bivalent alky l group. An “alkylene chain"’ is a polymethylene group, i.e.. -(CH2)_n-, wherein n is a positive integer, preferably from 1 to 6, from 1 to 4, from 1 to 3, from 1 to 2, or from 2 to 3. A substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are replaced with a substituent. Suitable substituents include those described below for a carbon atom.

[0020] The term “alkenylene’' refers to a bivalent alkeny l group. A substituted alkenylene chain is a polymethylene group containing at least one double bond in which one or more hydrogen atoms are replaced with a substituent. Suitable substituents include those described below for a carbon atom.

[0021] The terms “halogen” and “halo” are used interchangeably and mean F, Cl, Br, or I. [0022] The term “ary l” used alone or as part of a larger moiety as in “aralky l”, “aralkoxy”, or “aryloxyalkyl”, refers to monocyclic and bicyclic ring systems having a total of five to fourteen ring members, wherein each ring atom is carbon, at least one ring in the system is aromatic and wherein each ring in the sy stem contains three to seven ring members. The term “aryl” may be used interchangeably with the term “aryl ring”. In certain embodiments of the present disclosure, “aryl” refers to an aromatic ring system which includes, but not limited to. phenyl, biphenyl, naphthyl, anthracyl and the like, which may bear one or more substituents. Also included within the scope of the term “ary l”, as it is used herein, is a group in which an aromatic ring is fused to one or more non-aromatic carbocyclic rings.

[0023] The term “cycloalkyl ring” refers to a fully saturated carbocyclic ring.

[0024] A first ring is “spiro-fused” to a second ring when the two rings share a single ring carbon atom. An example of a spiro-fused ring sy stem is: ^H

[0025] A first ring is “fused” to a second ring when the two rings share two ring carbon atoms that are directly connected to one another. Example of fused ring sy stems are:

[0026] A first ring is “bridged” to a second ring when the two rings share two ring carbon atoms that are separated by at least one ring carbon atom. Example of bridged ring systems [0027] The terms “heteroaryl” and "heteroar-". used alone or as part of a larger moiety, e.g., "heteroaralkyl". or "heteroaralkoxy". refer to groups having 5 to 10 ring atoms, preferably 5, 6, or 9 ring atoms; having 6, 10, or 14 n electrons shared in a cyclic array; and having, in addition to carbon atoms, from one to five heteroatoms. The term “heteroatom” refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quatemized form of a basic nitrogen. Heteroaryl groups include, without limitation, thienyl, furanyl, pyrrolyl. imidazolyl, pyrazolyl, triazolyl, tetrazolyl. oxazolyl, isoxazolyl. oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl. The terms “heteroaryl” and “heteroar-”, as used herein, also include groups in which a heteroaromatic ring is fused to one or more aryl or heteroaryl rings such that the resulting bi- or multicyclic ring system as a whole is fully aromatic. Nonlimiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzothiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, and phenoxazinyl. A heteroaryl group may be mono- or bicyclic. The term “heteroaryl” may be used interchangeably with the terms “heteroaryl ring”, “heteroaryl group”, or “heteroaromatic”, any of which terms include rings that are optionally substituted. The term "heteroaralk l" refers to an alkyl group substituted by a heteroaryl, wherein the alkyl and heteroaryl portions independently are optionally substituted.

[0028] As used herein, the terms “heterocycle”, “heterocyclyl”, “heterocyclic radical”, and “heterocyclic ring” are used interchangeably and refer to a stable 5- to 7-membered monocyclic or 7- to 10-membered bicyclic heterocyclic moiety that is either saturated or partially unsaturated, and having, in addition to carbon atoms, one or more, preferably one to four, heteroatoms, as defined above. When used in reference to a ring atom of a heterocycle, the term “nitrogen” includes a substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur or nitrogen, the nitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or +NR (as in N-substituted pyrrolidinyl).

[0029] A heterocyclic ring can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure and any of the ring atoms can be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, without limitation, tetrahydrofuranyl, tetrahydrothiophenyl pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl. decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. The terms "heterocycle", “heterocyclyl”. "heterocycly l ring”, “heterocyclic group”, “heterocyclic moiety”, and “heterocyclic radical”, are used interchangeably herein, and also include groups in which a heterocyclyl ring is fused to one or more aryl, heteroaryl, or cycloaliphatic rings, such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, or tetrahydroquinolinyl. A heterocyclyl group may be mono- or bicyclic. The term "heterocyclylalkyl" refers to an alkyl group substituted by a heterocyclyl, wherein the alkyl and heterocyclyl portions independently are optionally substituted.

[0030] As described herein, compounds of the disclosure may contain “optionally substituted” moieties. In general, the term “substituted”, whether preceded by the term “optionally” or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent. Unless otherwise indicated, an “optionally substituted” group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position. Combinations of substituents envisioned by this disclosure are preferably those that result in the formation of stable or chemically feasible compounds. The term “stable”, as used herein, refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and, in certain embodiments, their recovery , purification, and use for one or more of the purposes disclosed herein.

[0031] The terms “cyano-substituted C_x-C_y alkyl” and “fluoro-substituted C_x-C_y alkyl” where each of x and y is an integer, refer to the corresponding alkyl having one or more of the indicated substituents in place of hydrogen.

[0032] Suitable monovalent substituents on a substitutable carbon atom of an “optionally -C(O)CH₂C(O)R°; -C(NOR°)R°; (CH₂)O-4SSR°; -(CH₂)O-4S(0)₂R°; -(CH₂)₀-4S(O)₂OR^O; -(CH₂)O-40S(0)₂R°; -S(0)₂NR^O ₂; (CH₂)O-4S(0)R°; N(R°)S(O)₂NR^O ₂; -N(R°)S(O)₂R^O; -N(OR°)R°; -C(NH)NR°₂; -P(O)₂R^O; P(O)R^O ₂; OP(O)R^O ₂; -OP(O)(OR^O)₂; SiR°₃; -(Ci-4 straight or branched alkylene)O-N(R°)₂; or -(C1-4 straight or branched alkylene)C(O)O- N(R°)₂, wherein each R° may be substituted as defined below and is independently hydrogen, C1-6 aliphatic, -CH₂Ph, -0(CH₂)o-iPh, -CH₂-(5-6 membered heteroaryl ring), or a 5-6- membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of R°, taken together with their intervening atom(s), form a 3-12- membered saturated, partially unsaturated, or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, which may be substituted as defined below.

[0033] Suitable monovalent substituents on R° (or the ring formed by taking two independent occurrences of R° together with their intervening atoms) are independently ( g y ) ( ) , where preceded by "halo’' is substituted only with one or more halogens, and is independently selected from C1-4 aliphatic, -CH₂Ph, -0(CH₂)o-iPh, or a 5-7-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents on a saturated carbon atom of R° include =0 and =S. [0034] Suitable divalent substituents on a saturated carbon atom of an “optionally substituted" group include the following: =0, =S, =NNR*₂, =NNHC(O)R*, =NNHC(O)OR*, =NNHS(O)₂R*, =NR*, =NOR*, -O(C(R*₂))_2.3O-, or -S(C(R*₂))_2.3S-, wherein each independent occurrence of R* is selected from hydrogen, C1-6 aliphatic which may be substituted as defined below, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents that are bound to vicinal substitutable carbons of an “optionally substituted” group include: -O(CR*₂)_2.3O-, wherein each independent occurrence of R* is selected from hydrogen, C1-6 aliphatic which may be substituted as defined below, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. [0035] Suitable substituents on the aliphatic group of R* include halogen, membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0036] Suitable substituents on a substitutable nitrogen of an "‘optionally substituted⁷’ group hydrogen, C1-6 aliphatic which may be substituted as defined below, unsubstituted -OPh, or a substituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of R taken together with their intervening atom(s) form an unsubstituted 3-12-membered saturated, partially unsaturated, or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0037] Suitable substituents on the aliphatic group and the substituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur of R^: are independently halogen, -R*, (haloR*), -OH, -OR*, - O(haloR*), -CN. -C(O)OH, -C(O)OR*. -NH₂, -NHR*, -NR*₂, or NO₂, wherein each R* is unsubstituted or where preceded by “halo” is substituted only with one or more halogens, and is independently C1-4 aliphatic, -CH₂Ph, -0(CH₂)o-iPh, or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0038] As used herein, the term “pharmaceutically acceptable derivative” means any nontoxic salt, ester, salt of an ester or other derivative of a compound of this disclosure that, upon administration to a recipient, is capable of providing, either directly or indirectly, a compound of this disclosure or an active metabolite or residue thereof.

[0039] As used herein, the term “pharmaceutically acceptable salt” refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein by reference. Pharmaceutically acceptable salts of the compounds of this disclosure include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3- phenylpropionate, phosphate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like.

[0040] Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N⁺(Ci-4alkyl)4 salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary' ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate and aryl sulfonate.

[0041] Unless otherwise stated, structures depicted herein are also meant to include all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational)) forms of the structure: for example, the R and S configurations for each asymmetric center, Z and E double bond isomers, rotational isomers (atropisomers) and Z and E conformational isomers. Therefore, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the disclosure. Unless otherwise stated, all tautomeric forms of the compounds of the disclosure are within the scope of the disclosure. Additionally, unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures including the replacement of hydrogen by deuterium or tritium, or the replacement of a carbon by a ¹³C- or ¹⁴C-enriched carbon are within the scope of this disclosure. Such compounds are useful, for example, as analytical tools, as probes in biological assays, or as therapeutic agents in accordance with the present disclosure. [0042] Combinations of substituents and variables envisioned by this disclosure are only those that result in the formation of stable compounds. The term “stable”, as used herein, refers to compounds which possess stability sufficient to allow manufacture and which maintains the integrity of the compound for a sufficient period of time to be useful for the purposes detailed herein (e.g., therapeutic or prophylactic administration to a subject).

[0043] The recitation of a listing of chemical groups in any definition of a variable herein includes definitions of that variable as any single group or combination of listed groups. The recitation of an embodiment for a variable herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof.

[0044] The term “biological sample”, as used herein, includes, without limitation, cell cultures or extracts thereof; biopsied material obtained from a mammal or extracts thereof; and hair, skin, blood, saliva, urine, feces, semen, tears, or other body fluids or extracts thereof. Inhibition of activity of a protein kinase, for example, WeelA kinase or a mutant thereof, or Mytlkinase or a mutant thereof., in a biological sample is useful for a variety of purposes that are known to one of skill in the art. Examples of such purposes include, but are not limited to, blood transfusion, organ transplantation, biological specimen storage, and biological assays.

[0045] As used herein, a “disease or disorder associated with WeelA kinase and Mytl kinase” or, alternatively, “a WeelA kinase-mediated and Mytl kinase-mediated disease or disorder” means any disease or other deleterious condition in which both WeelA kinase, or a mutant thereof, and Mytl kinase or a mutant thereof, are known or suspected to play a role.

[0046] The term “subject”, as used herein, means a mammal and includes human and animal subjects, such as domestic animals (e.g., horses, dogs, cats, etc.). The terms “subject” and “patient” are used interchangeably. In some embodiments, the “patient” or “subject” means an animal, preferably a mammal, and most preferably a human.

[0047] The term “pharmaceutically acceptable carrier, adj uvant, or vehicle” refers to a nontoxic carrier, adjuvant, or vehicle that does not destroy the pharmacological activity' of the compound with which it is formulated. Pharmaceutically acceptable carriers, adjuvants or vehicles that may be used in the compositions of this disclosure include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose- based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat. The amount of compounds of the present disclosure that may be combined with the carrier materials to produce a composition in a single dosage form will vary depending upon the host treated, the particular mode of administration, etc. Preferably, provided compositions are formulated so that a dosage of between 0.01 to about 100 mg/kg, or about 0.1 mg/kg to about 50 mg/kg, and preferably from about 1 mg/kg to about 25 mg/kg. of subject body weight/day of the inhibitor can be administered to a patient receiving these compositions to obtain the desired therapeutic effect. The amount of a compound of the present disclosure in the composition will also depend upon the particular compound in the composition.

[0048] As used herein, the terms “treatment,” “treat,” and “treating” refer to partially or completely alleviating, inhibiting, delaying onset of, preventing, ameliorating and/or relieving a disorder or condition, or one or more symptoms of the disorder or condition, as described herein. In some embodiments, treatment may be administered after one or more symptoms have developed. In some embodiments, the term “treating” includes preventing or halting the progression of a disease or disorder. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered to a susceptible individual prior to the onset of symptoms (e.g., in light of ahistory of symptoms and/or in light of genetic or other susceptibility factors). Treatment may also be continued after symptoms have resolved, for example to prevent or delay their recurrence. Thus, in some embodiments, the term “treating” includes preventing relapse or recurrence of a disease or disorder.

[0049] As used herein, the term “inhibitor” is defined as a compound that binds to and /or inhibits the target protein kinase with measurable affinity. In certain embodiments, an inhibitor has an IC50 and/or binding constant of less than about 50 pM, less than about 1 pM, less than about 500 nM, less than about 100 nM, less than about 50 nM, less than about 20 nM, or less than about 10 nM.

[0050] The terms “measurable affinity” and “measurably inhibit,” as used herein, means a measurable change in both WeelA kinase or Mytl kinase activity between a sample comprising a compound of the present disclosure, or composition thereof, and an equivalent sample comprising WeelA kinase or Mytl kinase, in the absence of said compound, or composition thereof. [0051] As used herein, the term “parenteral administration’" means subcutaneous, intravenous, intramuscular, intra-articular. intra-synovial, intrastemal, intrathecal, intrahepatic, intralesional and/or intracranial injection or infusion techniques. Weel Kinase Family

[0052] The Weel kinase family consists of three serine/threonine kinases sharing conserved molecular structures and encoded by the following genes: WEE1 (alternatively, WEE1 G2 checkpoint kinase or WeelA kinase), PKMYT1 (alternatively MYT1 kinase or membrane-associated tyrosine- and threonine-specific cdc2-inhibitory kinase), and WEE2 (alternatively WEE2 oocyte meiosis inhibiting kinase or WeelB kinase). In eukaryotic somatic cells, WeelA kinase and Mytl kinase play a key role in cell cycle regulation, in particular, in the entry into mitosis (Schmidt M, Rohs A, Platzer C, st al. Regulation of G2/M transition by inhibition of Weel and PMytI Kinases. Molecules. 2017;22:2045). Their role as regulators is crucial during normal cell cycle progression and in response to DNA damage as part of the DNA damage response (DDR) pathways.

[0053] Mytl kinase is a multi-functional protein kinase localized to the ER-Golgi complex that is known to play a regulatory role in the cell cycle by inhibiting Cdkl/cyclin Bl mediated mitosis (JY Zhu et al., J Med Chem. 2017; 60 (18), 7863-7875). As mentioned above and throughout, Mytl kinase inhibits the Cdkl/cyclin Bl activity through the phosphorylation of Tyrl5 and Thrl4 of Cdkl and sequestration of Cdkl from the nucleus. Additionally, Mytl kinase has been tied to orchestrating the ER-Golgi complex reassembly during mitotic exit.

[0054] WeelA kinase regulates entry into mitosis at the G2/M transition of the S phase by phosphorylating Tyrl of Cdkl to inactive the Cdkl/cyclin B complex. Cells with perturbed G1 checkpoint activity (e.g., cancer cells) rely on WeelA kinase to inhibit Cdkl to permit a G2/M arrest for DNA repair. If WeelA kinase activity is altered, a perturbed cell may enter mitosis prematurely without having the opportunity to fully replicate the entire DNA content or repair potential DNA lesions that might have occurred during S phase. This characterization of WeelA kinase’s role in the cell cycle has made it an attractive target for anticancer therapeutics, especially in combination with DNA-damaging agents (JY Zhu et al., J Med Chem. 2017; 60 (18), 7863-7875).

3. Compounds. [0055] The compounds of the present disclosure inhibit both WeelA kinase and PMytl kinase activities.

[0056] In some embodiments, the present disclosure provides a compound having structural formula I: solvate, enantiomer, tautomer, or diastereomer thereof, or a pharmaceutically acceptable salt of any of the foregoing wherein:

X is N or C(R):

Y is N or C(R);

Z is N or CH;

X and Y are not simultaneously N; each R is independently hydrogen, halo, or C₁-C₄ alkyl optionally substituted with halo;

R¹ is a 4-8 membered saturated heterocyclyl comprising one ring nitrogen atom and optionally comprising a second ring heteroatom selected from N, O, or S, wherein R¹ is optionally substituted on the one ring nitrogen atom with C₁-C₄ alkyl, wherein the optional Ci- C4 alkyl substituent is optionally further substituted with one or more substituents independently selected from halo, -OH, -SO₂-C₁-C₄ alkyl, and -O-C₁-C₄ alkyl; and

R² is C₁-C₄ alkyl, C2-C4 alkenyl, or C3-C4 cycloalkyl, wherein any one or more hydrogen atoms in the compound is optionally replaced with deuterium.

[0057] In some embodiments, the compound is a compound of formula II: having structural formula II: solvate, enantiomer, tautomer, or diastereomer thereof, or a pharmaceutically acceptable salt of any of the foregoing wherein: X is N or CH;

Y is N or CH;

X and Y are not simultaneously N; and

R³ is hydrogen or methyl.

[0058] As defined generally above and discussed throughout. Z is CH or N. In some embodiments, Z is CH. In some embodiments, Z is N.

[0059] As defined generally above and discussed throughout, in a compound of Formula I, each of X and Y is independently C(R) or N, wherein X and Y are not simultaneously N.

[0060] In some embodiments of a compound of Formula I, each of X and Y is C(R). In some embodiments of a compound of Formula I, X is N and Y is C(R). In some embodiments of a compound of Formula I. X is C(R) and Y is N.

[0061] As defined generally above and discussed throughout, in a compound of Formula I, each R is independently hydrogen, halo, or C₁-C₄ alky l optionally substituted with halo. In some embodiments of a compound of Formula I, each R is hydrogen. In some embodiments of a compound of Formula I where each of X and Y is C(R). one R is hydrogen and one R is methyl. In some embodiments of a compound of Formula I where each of X and Y is C(R), each R is methyl.

[0062] As defined generally above and discussed throughout, in a compound of Formula I, R¹ is a 4-8 membered saturated heterocyclyl comprising one ring nitrogen atom and optionally comprising a second ring heteroatom selected from N, O, or S, wherein R¹ is optionally substituted on the one ring nitrogen atom with C₁-C₄ alkyl optionally further substituted with one or more substituents independently selected from halo, -OH, -SO₂-C₁-C₄ alkyl, and -O-Ci- C4 alkyl.

[0063] In some embodiments of a compound of Formula I, R 1¹ is wherein R³ is hydrogen or C₁-C₄ alkyl optionally substituted with one or more substituents independently selected from halo, -OH, -SO₂-C₁-C₄ alkyl, and -O-C₁-C₄ alkyl. In some more specific embodiments of a compound of Formula I, R³ is hydrogen or methyl.

[0064] In some embodiments of a compound of Formula I, R¹ is, and in some embodiments of a compound of Formula II. the portion of the compound represented by is:

[0065] In some embodiments of a compound of Formula I, R¹ is azetidin-2-yl. azetidin-3- yl, morpholin-2-yl, morpholin-3-yl, thiomorpholin-2-yl, thiomorpholin-3-yl, piperazin-2-yl, piperazin-3-yl, azepan-3-yl, or azepan-5-yl, each optionally substituted with one or more methyl.

[0066] As defined generally above and discussed throughout in a compound of Formula I,

[0067] In some embodiments of a compound of Formula I, R² is C2-C4 alkenyl. In some embodiments of a compound of Formula I, R² is -CH2-CH=CH2. In some embodiments of a compound of Formula I, R² is methyl. In some embodiments of a compound of Formula I, R² is ethyl. In some embodiments of a compound of Formula I, R² is n-propyl. In some embodiments of a compound of Formula I, R² is cyclopropyl. In some embodiments of a compound of Formula I, R² is cyclobutyl.

[0068] In some embodiments, the compound of formula I may be a compound, or a pharmaceutically acceptable salt thereof, selected from Table 1.

Table 1. Exemplary Compounds

4. Uses, Formulation and Administration

Pharmaceutically Acceptable Compositions

[0069] According to another embodiment, the disclosure provides a composition comprising a compound of this disclosure or a pharmaceutically acceptable derivative thereof and a pharmaceutically acceptable carrier, adjuvant, or vehicle. The amount of compound in compositions of this disclosure is such that is effective to measurably inhibit both WeelA kinase, or a mutant thereof, and Mytl kinase or a mutant thereof in a biological sample or in a patient. In certain embodiments, a composition of this disclosure is formulated for administration to a patient in need of such composition. In some embodiments, a composition of this disclosure is formulated for oral administration to a patient.

[0070] Compositions of the present disclosure may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. Preferably, the compositions are administered orally, intraperitoneally or intravenously. Sterile injectable forms of the compositions of this disclosure may be aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example as a solution in 1,3 -butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer’s solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium.

[0071] For this purpose, any bland fixed oil may be employed including synthetic mono- or di-glycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, such as carboxymethyl cellulose or similar dispersing agents that are commonly used in the formulation of pharmaceutically acceptable dosage forms including emulsions and suspensions. Other commonly used surfactants, such as Tweens, Spans and other emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for the purposes of formulation.

[0072] In some embodiments, the compounds or compositions disclosed herein are administered orally. Pharmaceutically acceptable compositions of this disclosure may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use. carriers commonly used include lactose and com starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried cornstarch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added.

[0073] Alternatively, pharmaceutically acceptable compositions of this disclosure may be administered in the form of suppositories for rectal administration. These can be prepared by mixing the agent with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.

[0074] Pharmaceutically acceptable compositions of this disclosure may also be administered topically, especially when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs.

[0075] Topical application for the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topically -transdermal patches may also be used. [0076] For topical applications, provided pharmaceutically acceptable compositions may be formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers. Carriers for topical administration of compounds of this disclosure include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water. Alternatively, provided pharmaceutically acceptable compositions can be formulated in a suitable lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2ocfyldodecanol. benzy l alcohol and water.

[0077] For ophthalmic use, provided pharmaceutically acceptable compositions may be formulated as micronized suspensions in isotonic, pH adjusted sterile saline, or, preferably, as solutions in isotonic, pH adjusted sterile saline, either with or without a preservative such as benzalkonium chloride. Alternatively, for ophthalmic uses, the pharmaceutically acceptable compositions may be formulated in an ointment such as petrolatum.

[0078] Pharmaceutically acceptable compositions of this disclosure may also be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other conventional solubilizing or dispersing agents.

[0079] Most preferably, pharmaceutically acceptable compositions of this disclosure are formulated for oral administration.

[0080] The amount of compounds of the present disclosure that may be combined with the carrier materials to produce a composition in a single dosage form will vary depending upon the host treated, the particular mode of administration. Preferably, provided compositions should be formulated so that a dosage of betw een 0.001 - 100 mg/kg body weight/day of the inhibitor can be administered to a patient receiving these compositions.

[0081] It should also be understood that a specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the particular disease being treated. The amount of a compound of the present disclosure in the composition will also depend upon the particular compound in the composition. Uses of Compounds and Pharmaceutically Acceptable Compositions

[0082] Compounds and compositions described herein are generally useful for the inhibition of both WeelA kinase and Mytl kinase.

[0083] The activity of a compound utilized in this disclosure as an inhibitor of WeelA kinase and Mytl kinase, or a mutant of either of the foregoing, may be assayed in vitro, in vivo or in a cell line. In vitro assays include assays that determine inhibition of either the phosphorylation activity and/or the subsequent functional consequences, or ATPase activity of activated WeelA kinase, activated Mytl kinase, or a mutant of either of the foregoing. Alternate in vitro assays quantitate the ability of the inhibitor to bind to WeelA kinase and/or to Mytl kinase.

[0084] The provided compounds are useful for treating one or more disorders associated with activity of both Weel A kinase and Mytl kinase. Thus, in certain embodiments, the present disclosure provides a method for treating a WeelA kinase/Mytl kinase-mediated disorder comprising the step of administering to a patient in need thereof a compound of the present disclosure that is a dual inhibitor, or a pharmaceutically acceptable composition thereof.

[0085] As used herein, the term “WeelA kinase/Mytl kinase-mediated’’ disorder or condition as used herein means any disease or other deleterious condition in which both WeelA kinase and Mytl kinase, or a mutant of either or both of the foregoing, are known to play a role. Accordingly, another embodiment of the present disclosure relates to treating or lessening the severity of one or more diseases in which both WeelA kinase and Mytl kinase, or a mutant of either or both of the foregoing, are known to play a role. Specifically, the present disclosure relates to a method of treating or lessening the severity of a disease or condition selected from a proliferative disorder, wherein said method comprises administering to a patient in need thereof a compound or composition according to the present disclosure.

[0086] In some embodiments, the present disclosure provides a method of inhibiting both WeelA kinase and Mytl kinase activity in a subject comprising the step of administering to the subject an effective amount of a compound of the present disclosure that is a dual inhibitor, or a pharmaceutically acceptable composition.

[0087] In some embodiments, the present disclosure provides a method for treating or lessening the severity of one or more disorders selected from a cancer comprising the step of administering to the subject an effective amount of a compound, or a pharmaceutically acceptable composition thereof, of the present disclosure. In some embodiments, the cancer is associated with a solid tumor.

[0088] In some embodiments, the present disclosure provides a method of treating a subject suffering from a cancer or other disordered cell growth characterized by both aberrant Weel A kinase and aberrant Mytl kinase activity comprising the step of administering to the subject an effective amount of a compound of the present disclosure that is a dual inhibitor, or a pharmaceutically acceptable composition thereof. In some embodiments, aberrant WeelA kinase and aberrant Mytl kinase activity includes elevated activity, or overexpression, or undesirable activity as compared to a non-diseased state. In some such embodiments, aberrant WeelA kinase activity and aberrant Mytl kinase activity may include perturbed Cdkl activity', replication stress, altered mitosis, and DNA damage. In some embodiments, the subject to be treated has previously been treated with either a mono-specific Weel A kinase inhibitor or Mytl kinase inhibitor (neither of which is a dual inhibitor) and has developed resistance to or is refractory to such treatment. For example, such a subject could be resistant or refractory to the Mytl kinase inhibitor RP-6306, or the WeelA kinase inhibitors AZD1775, Debio0123 or ZnC3

[0089] In some embodiments, the cancer to be treated by a compound disclosed herein is selected from a brain cancer, a cervicocerebral cancer, a cardiac cancer, a gastrointestinal cancer, an esophageal cancer, a thyroid cancer, a small cell cancer, a non-small cell cancer, a breast cancer, a lung cancer, a stomach cancer, a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms' cancer, a skin cancer, malignant melanoma, a neuroblastoma, an osteosarcoma, an Ewing's tumor, a soft part sarcoma, an acute leukemia, a chronic lymphatic leukemia, a chronic myelocytic leukemia, polycythemia vera, a malignant lymphoma, multiple myeloma, a Hodgkin's lymphoma, and a non-Hodgkin’s lymphoma. In some embodiments, the subject is suffering from a cancer selected from a uterine serous carcinoma and a renal cancer.

[0090] In some embodiments, the breast cancer is selected from ductal carcinoma in situ (DCIS), invasive ductal carcinoma (IDC), lobular carcinoma in situ (LCIS), invasive lobular cancer (ILC), triple negative breast cancer (TNBC), inflammatory breast cancer (IBC), metastatic breast cancer (MBC), medullary carcinoma, tubular carcinoma, mucinous carcinoma (colloid), and Paget disease of the breast or nipple (commonly known as Paget disease). [0091] In some embodiments, the uterine cancer is selected from endometrial cancer and uterine sarcoma. In some embodiments, the uterine cancer is endometrial cancer. In some embodiments, the uterine cancer is uterine sarcoma.

[0092] In some embodiments, the ovarian cancer is selected from epithelial ovarian carcinomas, germ cell tumors, and stromal cell tumors.

[0093] In some embodiments, the stomach cancer is selected from adenocarcinoma, lymphoma, gastrointestinal stromal tumors (GISTs), carcinoid tumors, and hereditary (familial) diffuse gastric cancer.

[0094] In some embodiments the esophageal cancer is selected from squamous cell carcinoma, small cell carcinoma, and adenocarcinoma. In some embodiments the esophageal cancer is selected from squamous cell carcinoma and adenocarcinoma. In some embodiments, the esophageal cancer is squamous cell carcinoma. In some embodiments, the esophageal cancer is adenocarcinoma.

[0095] In some embodiments, the lung cancer is selected from non-small cell lung cancer, lung nodules, small cell lung cancer, and mesothelioma. In some embodiments, the lung cancer is non-small cell lung cancer.

[0096] In some embodiments the colorectal cancer is selected from adenocarcinoma, gastrointestinal stromal tumors (GIST), lymphoma, carcinoids, Turcot syndrome, Peutz- Jeghers syndrome (PJS), familial colorectal cancer (FCC), and juvenile polyposis coli.

[0097] In some embodiments, the cancer is associated with deregulation of cyclin El. In some embodiments, the cancer associated with deregulation of cyclin El is ovarian cancer.

[0098] In some embodiments, the cancer is associated with deregulation of p53. In some embodiments, the cancer associated with deregulation of p53 is selected from a brain cancer, a cervicocerebral cancer, a cardiac cancer, a gastrointestinal cancer, an esophageal cancer, a thyroid cancer, a small cell cancer, a non-small cell cancer, a breast cancer, a lung cancer, a stomach cancer, a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms' cancer, a skin cancer, malignant melanoma, a neuroblastoma, an osteosarcoma, an Ewing's tumor, a soft part sarcoma, an acute leukemia, a chronic lymphatic leukemia, a chronic myelocytic leukemia, polycythemia vera, a malignant lymphoma, multiple myeloma, a Hodgkin's lymphoma, and a non-Hodgkin's lymphoma. In some such embodiments, the cancer associated with deregulation of p53 is selected from uterine serous carcinoma and a renal cancer. [0099] In some embodiments, the cancer is associated with deregulation of Cdkl. In some embodiments, the cancer associated with deregulation of Cdkl is selected from a brain cancer, a cervi cocerebral cancer, a cardiac cancer, a gastrointestinal cancer, an esophageal cancer, a thyroid cancer, a small cell cancer, a non-small cell cancer, a breast cancer, a lung cancer, a stomach cancer, a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms' cancer, a skin cancer, malignant melanoma, a neuroblastoma, an osteosarcoma, an Ewing's tumor, a soft part sarcoma, an acute leukemia, a chronic lymphatic leukemia, a chronic myelocytic leukemia, polycythemia vera, a malignant lymphoma, multiple myeloma, a Hodgkin's lymphoma, and a non-Hodgkin’s lymphoma. In some such embodiments, the cancer associated with deregulation of Cdkl is selected from uterine serous carcinoma and a renal cancer.

[0100] In some embodiments, the cancer is associated with deregulation of Cdk2. In some embodiments, the cancer associated with deregulation of Cdk2 is selected from a brain cancer, a cervicocerebral cancer, a cardiac cancer, a gastrointestinal cancer, an esophageal cancer, a thyroid cancer, a small cell cancer, a non-small cell cancer, a breast cancer, a lung cancer, a stomach cancer, a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms' cancer, a skin cancer, malignant melanoma, a neuroblastoma, an osteosarcoma, an Ewing's tumor, a soft part sarcoma, an acute leukemia, a chronic lymphatic leukemia, a chronic myelocytic leukemia, polycythemia vera, a malignant lymphoma, multiple myeloma, a Hodgkin's lymphoma, and a non-Hodgkin’s lymphoma. In some such embodiments, the cancer associated with deregulation of Cdkl is selected from uterine serous carcinoma and a renal cancer.

[0101] Depending upon the particular condition, or disease, to be treated, additional therapeutic agents, which are normally administered to treat that condition, may also be present in the compositions of this disclosure. As used herein, additional therapeutic agents that are normally administered to treat a particular disease, or condition, are known as “appropriate for the disease, or condition, being treated.”

[0102] For example, compounds of the present disclosure, or a pharmaceutically acceptable composition thereof, are administered in combination with chemotherapeutic agents to treat proliferative diseases and cancer. Examples of known chemotherapeutic agents include, but are not limited to, Adriamycin, dexamethasone, vincristine, cyclophosphamide, fluorouracil, topotecan, taxol, interferons, platinum derivatives, taxane (e.g., paclitaxel), vinca alkaloids (e.g., vinblastine), anthracy clines (e.g., doxorubicin), epipodophyllotoxins (e.g., etoposide), cisplatin, an mTOR inhibitor (e.g., a rapamycin), methotrexate, actinomycin D, dolastatin 10, colchicine, emetine, trimetrexate, metoprine, cyclosporine, daunorubicin, teniposide, amphotericin, alkylating agents (e.g., chlorambucil), 5 -fluorouracil, camptothecin, cisplatin, metronidazole, and Gleevec™, among others. In other embodiments, a compound of the present disclosure is administered in combination with a biologic agent, such as Avastin or VECTIBIX.

[0103] In some embodiments, compounds of the present disclosure or a pharmaceutically acceptable composition thereof, are administered in combination with an agent selected from fasudil, sirolimus, imatinib, gefitinib, erlotinib, sorafenib, sunitinib, dasatinib, lapatinib, nilotinib, temsirolimus, everolimus, pazopanib, ruxolitinib, vandetanib, vemurafenib, crizotinib, icotinib, axitinib, tofacitinib, bosutinib, cabozantinib, ponatinib, regorafenib, afatinib, dabrafenib, trametinib, ibrutinib, nintedanib, idelalisib. ceritinib, apatinib rivoceranib, ripasudil. alectinib, cobimetinib, lenvatimb, palbociclib, radotinib. osimertinib. olmutinib. neratinib, ribociclib, copanlisib, abemaciclib, acalabrutinib, midostaurin, brigatinib, baricitinib, netarsudil, tivozanib, simotinib, fostamatinib, encorafenib, binimetinib, catequentinib, duvelisib, dacomitinib, lorlatinib, larotrectinib, gilteritinib, pyrotinib, fruquintinib, erdafitinib, alpelisib. umbralisib. leniolisib, pexidartinib, entrectinib, upadacitinib, fedratinib, zanubrutinib, flumatinib, peficitinib, delgocitinib, avapritinib, selumetinib, tucatinib, pemigatinib, capmatinib tabrecta, selpercatinib, ripretinib, tirabrutinib, almonertinib, pralsetinib, filgotinib, tirbanibulin, orelabrutinib, tepotinib, and trilaciclib. See List of clinically approved kinase inhibitors \ MRC Protein Phosphorylation Ubiquitylation Unit available at www.ppu.mrc.ac.uk/list-clinically-approved-kinase-inhibitors, incorporated herein by reference in its entirety.

[0104] In certain embodiments, compounds of the present disclosure, or a pharmaceutically acceptable composition thereof, are administered in combination with an antiproliferative or chemotherapeutic agent selected from any one or more of abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, amifostine, anastrozole, arsenic trioxide, asparaginase, azacitidine, BCG Live, bevacizumab, fluorouracil, bexarotene, bleomycin, bortezomib, busulfan, calusterone, capecitabine, camptothecin, carboplatin, carmustine, celecoxib, cetuximab, chlorambucil, cladribine. clofarabine, cyclophosphamide, cytarabine, dactinomycin, darbepoetin alfa, daunorubicin, decitabine, denileukin, dexrazoxane, docetaxel, doxorubicin (neutral), doxorubicin hydrochloride, dromostanolone propionate, epirubicin, epoetin alfa, erlotinib, estramustine, etoposide phosphate, etoposide, exemestane. filgrastim, floxuridine fludarabine, fulvestrant, gefitinib, gemcitabine, gemtuzumab, goserelin acetate, histrelin acetate, hydroxyurea, ibritumomab, idarubicin, ifosfamide, imatinib mesylate, interferon alfa-2a, interferon alfa-2b, irinotecan, lenalidomide, letrozole, leucovorin, leuprolide acetate, levamisole, lomustine, megestrol acetate, melphalan, mercaptopurine. 6-MP, mesna, methotrexate, methoxsalen, mitomycin C, mitotane. mitoxantrone, nandrolone, nelarabine. nofetumomab, oprelvekin, oxaliplatin, paclitaxel, palifermin, pamidronate, pegademase, pegaspargase, pegfilgrastim, pemetrexed disodium, pentostatin, pipobroman, plicamycin, porfimer sodium, procarbazine, quinacrine, rasburicase, rituximab, sargramostim, sorafenib, streptozocin, sunitinib maleate, talc, tamoxifen, temozolomide, teniposide, VM-26, testolactone, thioguanine, 6-TG, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin, ATRA, uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine, zoledronate, or zoledronic acid.

[0105] In certain embodiments, compounds of the present disclosure, or a pharmaceutically acceptable composition thereof, are co-administered with a pharmaceutically acceptable DNA damaging agent.

[0106] In certain embodiments, compounds of the present disclosure, or a pharmaceutically acceptable composition thereof, are co-administered with radiation.

[0107] In certain embodiments, compounds of the present disclosure, or a pharmaceutically acceptable composition thereof, are administered in combination with a monoclonal antibody or an siRNA therapeutic.

[0108] In certain embodiments, compounds of the present disclosure, or a pharmaceutically acceptable composition thereof, are administered in combination with a targeted therapy selected from (i) an inhibitor of a kinase selected from MET, MEK, mTOR, FLT3, BRAF, KIT, PDGFR, FDFR, PEK, EGFR, AKT, and KRAS, (ii) an inhibitor of a fusion kinase like BCR-ABL, ALK, RET and ROS, JAK, CDK4/6, and KRAS, (iii) epigenetic modulators such as an HD AC inhibitor, (iv) immuno-oncology agents such as those targeting PD1, PDL1, and CTLA4, (v) antibody drug conjugates such as those targeting Her2, CD38, BCMA. CD19. nectin i. trop2, CD79, and CD22, (vi) bispecific T cell engagers (BiTEs), (vii) transcription factor modulators such as those targeting IKZF (i.e., IMiDs, and EZH2), (viii) steroid receptor modulators such as those targeting AR and ER, and (ix) proteasome inhibitors such as bortezomib, ixazomib. carfilzomib and (x) agents targeting apoptosis such as inhibitors of BCL-2, BCL-XL, MCL1, IAP, or TRAIL/Death Receptor agonists. [0109] In some embodiments, compounds of the present disclosure, or a pharmaceutically acceptable composition thereof, are administered in combination with an inhibitor of a DNA repair protein other than WeelA kinase or Mytl kinase. Such inhibitors include those that inhibit one or more of the following CHK1, CHK2, ATM, ATR, Pol Theta, CDC7, DNAPK, PLK1, WRN, PARP and Aurora A/B.

[0110] Those additional agents may be administered separately from an inventive compound-containing composition, as part of a multiple dosage regimen. Alternatively, those agents may be part of a single dosage form, mixed together with a compound of this disclosure in a single composition. If administered as part of a multiple dosage regime, the two active agents may be submitted simultaneously, sequentially or within a period of time from one another, for example, within one, two, three, four, five, six, seven, eight, nine, ten, eleven, or twelve hours from one another.

[0111] As used herein, the term “combination,” “combined,” “co-administered” and related terms refer to the simultaneous or sequential administration of therapeutic agents in accordance with this disclosure. For example, a compound of the present disclosure may be administered with another therapeutic agent simultaneously or sequentially in separate unit dosage forms or together in a single unit dosage form. Accordingly, the present disclosure provides a single unit dosage form comprising a provided compound, an additional therapeutic agent, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

[0112] The amount of both, an inventive compound and additional therapeutic agent (in those compositions which comprise an additional therapeutic agent as described above) that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. Preferably, compositions of this disclosure should be formulated so that a dosage of between 0.001 - 100 mg/kg body weight/day of an inventive can be administered.

[0113] In those compositions which comprise an additional therapeutic agent, that additional therapeutic agent and the compound of this disclosure may act synergistically. Therefore, the amount of additional therapeutic agent in such compositions will be less than that required in a monotherapy utilizing only that therapeutic agent. In such compositions a dosage of between 0.001 - 1,000 pg/kg body weight/day of the additional therapeutic agent can be administered.

[0114] The amount of additional therapeutic agent present in the compositions of this disclosure will be no more than the amount that would normally be administered in a composition comprising that therapeutic agent as the only active agent. Preferably the amount of additional therapeutic agent in the presently disclosed compositions will range from about 50% to 100% of the amount normally present in a composition comprising that agent as the only therapeutically active agent.

[0115] In some embodiments, the compounds of the present invention may form a proteolysis targeting chimera (PROTAC) by being covalently linked directly or through a chemical linker to a small molecule capable of engaging an E3 ubiquitin ligase.

[0116] In some embodiments, the compounds of the present invention may form an antibody-drug conjugate (ADC) by being covalently linked directly or through a chemical linker to an antibody or an antigen-binding fragment of an antibody.

[0117] In some embodiments, the present disclosure provides a method for inhibiting both Weel A kinase and Mytl kinase in a biological sample.

[0118] In some embodiments, the present disclosure provides a method for inhibiting both Weel A kinase and Mytl kinase in vitro. In some such embodiments, the amount of Weel A kinase and Mytl kinase inhibition are both assessed based on a competitive ATP -binding assay. [0119] In some embodiments, the present disclosure provides a method for assessing Cdkl phosphorylation in a cell, comprising contacting said cell with a compound described herein. In one embodiment, the contacting step comprises incubating a cell with a compound presented herein. In some such embodiments, the cell is incubated for at least 4 hours. In some embodiments, the cell may comprise a DAOY medulloblastoma cell. In some embodiments, the cell may comprise a ACHN renal carcinoma cell. In some embodiments, the cell may comprise an A427 lung cancer cell. In some embodiments, the cell may comprise an OVCAR3 ovarian cancer cell.

EXAMPLES

[0120] General Methods

[0121] Reagents and solvents were purchased from commercial suppliers and used as received unless otherwise noted. Solvents were dried over molecular sieves 4A. Reactions were stirred using magnetic stir bars in glass vials or round bottomed flasks and heated using stirring plates. Solvents were removed on rotary evaporators, vacuum centrifuge, or by freeze-drying. Reaction progress was monitored by LC-MS or thin layer chromatography (TLC). Aluminium-backed TLC plates (6OF254) were visualized by UV-light (254 nm).

[0122] ¹H NMR and ¹³C NMR spectra were recorded on a 500 MHz ( ¹H NMR at 500 Spectra were processed using commercial software.

[0123] LC-MS were acquired on instruments using reversed phase C18 columns eluting with acetonitrile and water (containing 0.1% TFA or 0.03% ammonia) with mass spectrometers operating in ES (+ or -) ionization mode.

[0124] Flash chromatography was performed on silica gel or Cl 8 functionalized silica were performed on an automated flash purification system equipped with a diode array detector (200-400 nm), eluting with gradients of ethyl acetate and petroleum ether or methanol and dichloromethane (containing 0.03 % of ammonia.

[0125] Purity analyses were performed by HPLC reversed phase C18 columns eluting with acetonitrile and water (containing 0.1% TFA or 0.03% ammonia). UV -traces were recorded at 220 nm. Purifications by preparative HPLC were performed using reversed phase Cl 8 columns eluting with acetonitrile and water (containing 0.1% TFA or 0.03% ammonia).

[0126] Abbreviations: DMSO dimethylsulfoxide

Et ethyl

EtOAc ethyl acetate

EtOH ethanol

GC-MS gas chromatography- mass spectrometry

LDA lithium diisopropylamide mCPBA meta-chloroperbenzoic acid

MeOH methanol

MTBE methyl tert-butyl ether

NCS A-chlorosuccinimide

NBS A-bromosuccinimide

NMP A-methylpyrrolidine

LC-MS or LCMS liquid chromatography- mass spectroscopy

PdCh x dppf 1,1 -bis(diphenylphosphino)ferrocene palladium(II)dichloride pTSA p-lohienesul Ionic acid RT room temperature, normally 20 to 22 °C

TBAF tetrabutyl ammonium fluoride

TBS tert-Butyldimethylsilyl

TBSC1 tert-Butyldimethylsilyl chloride

TEA triethylamine

TFA trifluoroacetic acid

TFAA trifluoroacetic acid anhydride

THF tetrahydrofuran

TBME tert-butyl methyl ether

TLC thin layer chromatography t or Tret retention time

Triflic anhydride trifluoromethanesulfonic anhydride (TfzO)

UPLC Ultra high performance liquid chromatography

[0127] Example 1. Synthesis of 6-[(l-methylindazol-5-yl)amino]-2-(prop-2-en-l-yl)- l-{l-[(3S)-pyrrolidin-3-yl]pyrrolo[2,3-b]pyridin-6-yl}pyrazolo[3,4-d]pyrimidin-3-one (Compound 100)

[0128] tert-butyl (3S)-3-{6-bromopyrrolo[2.3-bJpyridin-l-yl}pyrrolidine-l-carboxylate [0129] (Tributylphosphoranylidene)acetonitrile (0.8 M in toluene, 1.14 ml, 0.91 mmol, 1.8 equiv.) was added to a solution of 6-bromo-lH-pyrrolo[2.3-b]pyridine (100 mg, 0.51 mmol, 1.0 equiv.) and tert-Butyl (R)-3-hydroxy-l-pyrrolidinecarboxylate (95 mg, 0.51 mmol, 1.0 equiv.) in anhydrous toluene (10 ml) under argon and the resulting brown solution was heated at 90 °C in a closed vial. The reactions were monitored by LCMS and additional reagents were added as needed to achieve full conversion of the aza-indole. After 21 h, a second portion of the alcohol tert-butyl (R)-3-hydroxy-l-pyrrolidinecarboxylate (48 mg. 0.26 mmol, 0.50 equiv.) and phosphorane (tributylphosphoranylidene)acetonitrile solution (570 pl, 0.46 mmol, 0.90 equiv.) were added and the reaction was continued at 100 °C for 6 h at which point LCMS indicated >95% conversion of the azaindole. The reaction mixture was cooled to RT and concentrated to give a brown oil, which was purified by flash chromatography on silica, eluting with a gradient of ethyl acetate in petroleum ether. The pure fractions were concentrated to give the title compound (166 mg, 89%) as a colorless oil. LCMS m/z found 310.0.

[0130] tert-butyl-(3S)-3-{6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)pyrazolo[3,4- djpyrimidin-l- yljpyrrolo[2.3-b Jpyridin-l-yl}pyrrolidine-l -carboxylate

[0131] A suspension of tert-butyl (3S)-3-{6-bromopyrrolo[2,3-b]pyridin-l-yl}pyrrolidine-l- carboxylate (166 mg, 0.45 mmol, 1.0 equiv.), 6-(methylsulfanyl)-2-(prop-2-en-l-yl)-lH- pyrazolo[3,4-d]pyrimidin-3-one (101 mg, 0.45 mmol, 1.0 equiv.), and cesium carbonate (443 mg, 1.36 mmol, 3.0 equiv.) in anhydrous 1,4-dioxane (7 ml) was degassed by argon bubbling for a few minutes. Cuprous iodide (104 mg, 0.54 mmol, 1 .2 equiv.) was added, followed by trans-l,2-bis(methylamino)cyclohexane (72 pl, 0.45 mmol, 1.0 equiv.) and the stirred mixture was heated at 90 °C in a closed vial under argon The reaction was monitored by LCMS. After 6 h, an additional portion of cuprous iodide (52 mg, 0.27 mmol, 0.60 equiv.) and trans- l,2-bis(methylamino)cy cl ohexane (36 pl, 0.23 mmol, 0.50 equiv.) were added and the reaction was continued for another 3 h, at which point LCMS indicated full conversion of the starting materials. The mixture was diluted with EtOAc (30 ml) and washed with dilute aqueous ammonia (30 ml) followed by saturated brine (30 ml). The organic phase was filtered through a phase-separator and concentrated under reduced pressure to give the crude compound (266 mg) as a brown solid which was used in the next step without further purification. LCMS m/z found 408.0, MH⁺ -Boc.

[0132] 6-[(l-methylindazol-5-yl)amino]-2-(prop-2-en-l-yl)-l-{l-[(3S)-pyrrolidin-3- yl]pyrrolo[2, 3-b ]pyridin-6-yl}pyrazolo[3, 4-d]pyrimidin-3-one [0133] mCPBA (75%, 145 mg, 0.63 mmol, 1.4 equiv.) was added to a stirred solution of tertbutyl (3S)-3-{6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)pyrazolo[3.4-d]pyrimidin-l- yl]pyrrolo[2,3-b]pyridin-l-yl}pyrrolidine-l-carboxylate (266 mg, ca. 85% purity, 0.45 mmol) in DCM (5 ml) at RT. Upon full conversion of the methylthio-pyrimidine starting material as judged by LCMS (15 min), l-methylindazol-5 -amine (77 mg, 0.52 mmol, 1.2 equiv.) was added and the resulting solution was heated in a closed vial at 30-40 °C until full conversion of the intermediates was observed (20 h. LCMS). The reaction mixture was cooled to RT, trifluoroacetic acid (1 ml, excess) was added, and the solution was stirred until the Boc-amine intermediate was consumed (15 min, LCMS). The mixture was partitioned between EtOAc and sat. brine (adjusted to pH ca. 12 with aq. NaOH). The organic phase was fdtered through a phase-separator and concentrated under reduced pressure to give the crude intermediate (210 mg) as a red solid. Part of this material (27 mg) was purified by reversed phase chromatography (Gemini NX-C18, 21x150 mm, water (0.1% TFA)/acetonitrile, gradient over 12 minutes, 25 ml/min) and the pure fractions were lyophilized to give the title compound (TFA salt, 15.5 mg, 43%) as a pale yellow solid. LCMS m/z found 507.0.

[0134] 'H NMR (400 MHz, dmso) 6 10.38 (s. 1H), 9.01 (d. J= 58.1 Hz, 2H), 8.90 (s, 1H). 8.45 - 8.21 (m, 2H), 7.99 (s, 1H), 7.79 (d, J = 3.2 Hz, 1H), 7.71 - 7.54 (m, 3H), 6.71 (d, J = 3.5 Hz, 1H), 5.80 - 5.64 (m, 1H), 5.57 - 5.42 (m, 1H), 5.03 (d, J= 10.3 Hz, 1H), 4.88 (d, J = 17.0 Hz, 1H), 4.64 (s, 2H). 4.02 (s, 3H), 3.78 - 3.69 (m, 2H), 3.58 - 3.51 (m, 1H), 3.48 - 3.35 (m. 2H), 2.38 - 2.28 (m. 1H).

[0135] Example 2. Synthesis of 6-[(l-methylindazol-5-yI)amino]-l-[l-(piperidin-4- yl)py rrolo [2,3-b] pyridin-6-yl] -2-(prop-2-en-l-yl)pyrazolo [3,4-d] pyrimidin-3-one (Compound 101)

[0136] tert-butyl 4- {6-bromopyrrolo[2,3-b]pyridin-l-yl}piperidine-l -carboxylate [0137] tert-butyl 4-[(4-methylbenzenesulfonyl)oxy]piperidine-l -carboxylate (700 mg, 1.97 mmol, 1.0 equiv.) was added to a stirred suspension of 6-bromo-lH-pyrrolo[2,3-b]pyridine (388 mg, 1.97 mmol, 1.0 equiv.) and cesium carbonate (1.28 g, 3.94 mmol, 2.0 equiv.) in anhydrous DMF (10 ml) at RT. After 2 days, the mixture was heated at 80 °C and additional tert-butyl 4-[(4-methylbenzenesulfonyl)oxy]piperidine-l-carboxylate (370 mg. 1.04 mmol, 0.53 equiv.) was added in portions over 8 h. After cooling to RT, the mixture was diluted with EtOAc (70 ml) and washed with 15% aq NaCl. (4x30 ml) followed by sat. brine (30 ml). The organic phase was filtered through a phase-separator and concentrated to a yellow oil, which was purified flash chromatography on silica, eluting with a gradient of 0-2.5% methanol in dichloromethane. The pure fractions were concentrated to give the title compound (154 mg, 21%) as a colorless oil. Additional product was isolated as a mixture with unreacted starting material. LCMS m/z found 324.0.

[0138] tert-butyl-4-{6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)pyrazolo[3,4- d]pyrimidin-l-yl]pyrrolo[2,3-b]pyridm-l-yl}piperidine-l-carboxylate

[0139] Prepared using a method similar to that employed in the second step of Example 1 using terl-butyl 4-{6-bromopyrrolo[2,3-b]pyridin-l-yl}piperidine-l-carboxylate (153 mg) and N,N'-dimethylethylenediamine as the ligand. Yield: 144 mg (69%) as a colorless oil. LCMS m/z found 422.0, MH⁺ -Boc.

[0140] 6-[(l-methylindazol-5-yl)amino]-l-[l-(piperidin-4-yl)pyrrolo[2,3-b]pyridin-6-yl]-2- (prop-2-en-l-yl)pyrazolo[3, 4-d]pyrimidin-3-one; trifluoroacetic acid

[0141] Prepared using a method similar to that employed in the third step of Example 1 using tert-butyl 4-{6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)pyrazolo[3,4-d]pyrimidin-l- yl]pyrrolo[2,3-b]pyridin-l-yl}piperidine-l-carboxylate (144 mg) to give the crude free base intermediate (183 mg) as a brown solid of which 20% was purified by prep-HPLC. Yield: 21 mg TFA salt (60%) as a white powder. LCMS m/z Found: 521.0 MH⁺.

[0142] ¹H NMR (400 MHz, dmso) 6 10.36 (s. 1H), 8.90 (s. 1H), 8.71 (d. J = 10.1 Hz, 1H). 8.44 - 8.17 (m, 3H), 7.99 (s, 1H), 7.61 (d, J = 18.6 Hz, 4H), 6.68 (d, J = 3.5 Hz, 1H), 5.72 (ddt, J = 16.2, 10.8, 5.7 Hz, 1H), 5.00 (d, J = 10.3 Hz, 1H), 4.93 (dq, J = 14.7, 5.0 Hz, 1H), 4.85 (d, J = 17.2 Hz, 1H), 4.66 (s, 2H), 4.02 (s, 3H). 3.44 (d, J = 12.7 Hz, 2H), 3.22 (q, J = 10.1 Hz, 2H), 2.27 - 2.06 (m, 4H).

[0143] Example 3. Synthesis of 6-[(l-methylindazol-5-yI)amino]-l-[l-(piperidin-4- yl)pyrazolo[3,4-b]pyridin-6-yl]-2-(prop-2-en-l-yl)pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid (Compound 102)

[0144] tert-butyl 4-{ 6-iodo-lH-pyrazolo[3, 4-b ]pyridin-l-yl}piperidine-l -carboxylate

[0145] Prepared using a method similar to that employed in the first step of Example 1 using 6-iodo-lH-pyrazolo[3,4-b]pyridine (100 mg, 0.41 mmol) and tert-butyl (3R)-3- hydroxypiperidine-1 -carboxylate (1.5 equiv ). Yield: 152 mg (87%) as a white crystalline solid. LCMS: found 451.0, MH⁺.

[0146] tert-butyl-4-{6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH,2H,3H-pyrazolo[3,4- d]pyrimidin-l-yl ]-lH-pyrazolo[ 3, 4-b ]pyridin-l-yl}piperidine-l -carboxylate

[0147] Prepared using a method similar to that employed in the second step of Example 1 using tert-butyl 4-{6-iodo-lH-pyrazolo[3,4-b]pyridin-l-yl}piperidine-l-carboxylate (150 mg) and N,N'-Dimethylethylenediamine as the ligand. Yield: 149 mg (81%) as a colorless film. LCMS m/z found 423.0, MH⁺-Boc. [0148] 6-[( 1 -methylindazol-5-yl)amino ]-l-[l-(piperidin-4-yl)pyrazolo[3, 4-b ]pyridin-6-yl ]-2- (prop-2-en-l-yl)pyrazolo[3, 4-d]pyrimidin-3-one; trifluoroacetic acid

[0149] Prepared using a method similar to that employed in the third step of Example 1 using tert-butyl 4-{6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH,2H,3H-pyrazolo[3,4- d]pyrimidin-l-yl]-lH-pyrazolo[3,4-b]pyridin-l-yl}piperidine-l-carboxylate (149 mg) to give the crude free base intermediate (122 mg) as a pink solid of which 20% was purified by prep- HPLC. Yield: 19.4 mg TFA salt (54%) as a pale-yellow powder. LCMS m/z Found: 522.2, MH .

[0150] ¹H NMR (400 MHz, dmso) 5 10.49 (s, 1H), 8.93 (s, 1H), 8.72 (s, 1H), 8.55 (d, J = 8.3 Hz, 1H), 8.44 (s, 1H), 8.28 (s, 2H), 8.08 (s, 1H), 7.90 (d, J = 8.6 Hz, 1H), 7.62 (s, 2H), 5.72 (ddt, J = 16.5, 11.1, 5.9 Hz, 1H). 5.15 (d, J = 11.9 Hz. 1H), 5.00 (d. J = 10.2 Hz, 1H), 4.86 (d, J = 17.1 Hz, 1H), 4.76 (d, J = 6.1 Hz, 2H), 4.04 (s, 3H), 3.26 (d, J = 11.8 Hz, 3H), 2.32 (d, J = 11.0 Hz, 2H), 2.14 (d, J = 13.4 Hz, 2H).

[0151] Example 4. Synthesis of 6-[(l-methyl-lH-indazol-5-yl)amino]-l-{l-[(3S)- piperidin-3-yl] - IH-pyrazolo [3, 4-b ] py ridin-6-yl }-2-(p ro p-2-en- l-yl)-lH,2H,3H- pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid (Compound 103).

[0152] tert-butyl (3S)-3-{6-iodo- 1 H-pyrazolo 3, 4-b ] 'pyridin- 1 -yl} piperidine- 1 -carboxylate [0153] Prepared using a method similar to that employed in the first step of Example 1 using 6-iodo-lH-pyrazolo[3,4-b]pyridine (100 mg, 0.41 mmol) and tert-butyl (3R)-3- hydroxypiperidine-1 -carboxylate (2.5 equiv.). Yield: 46 mg (26%) as a colorless oil. LCMS: found 372.8, MH⁺ -C₄H₈.

[0154] tert-butyl (3S)-3-{6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH,2H,3H- pyrazolo[3, 4-d ]pyrimidin- 1 -yl ]- lH-pyrazolo[3, 4-b ] pyridin- 1 -yl}piperidine- 1 -carboxylate [0155] Prepared using a method similar to that employed in the second step of Example 1 using tert-butyl (3S)-3- {6-iodo-lH-pyrazolo[3,4-b]pyridin- 1-yl [piperidine- 1 -carboxylate (150 mg) and N,N'-Dimethylethylenediamine as the ligand. The crude material was used in the next step without purification.

[0156] 6-[(l-methyl-lH-indazol-5-yl)amino]-l-{l-[(3S)-piperidin-3-yl]-lH-pyrazolo[3,4- b]pyridin-6-yl}-2-(prop-2-en-l-yl)-lH,2H.3H-pyrazolo[3.4-d]pyrimidin-3-one; trifluoroacetic acid

[0157] Prepared using a method similar to that employed in the third step of Example 1 using tert-butyl (3S)-3-{6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH,2H,3H-pyrazolo[3,4- d]pyrimidin-l-yl]-lH-pyrazolo[3,4-b]pyridin-l-yl}piperidine-l-carboxylate (crude) to give the crude free base intermediate (56 mg) as a yellow film of which 20% was purified by prep- HPLC. Yield: 5.8 mg TFA salt as a yellow powder. LCMS m/z Found: 522.0, MH⁺.

[0158] 1H NMR (400 MHz, dmso) 8 10.48 (s, 1H), 8.93 (s, 1H), 8.50 (s, 1H), 8.28 (s, 1H), 8.21 (s, 1H), 8.07 (s, 1H), 7.87 (d, J = 8.6 Hz, 1H), 7.62 (s, 2H), 5.73 (ddt, J = 16.7, 11.2, 5.9 Hz, 1H), 5.01 (d, J = 10.3 Hz, 1H), 4.86 (d, J = 17.1 Hz, 1H), 4.75 (d, J = 6.2 Hz, 3H), 4.04 (s, 3H), 3.18 - 2.85 (m, 4H), 2.22 - 1.95 (m, 2H), 1.77 (d, J = 13.0 Hz, 1H), 1.61 (d, J = 12.6 Hz. 1H). NH not observed.

[0159] Example 5. Synthesis of 6-[(l-methylindazol-5-yI)amino]-2-(prop-2-en-l-yl)-l-{l- [(3R)-pyrrolidin-3-yl]pyrrolo[2,3-b]pyridin-6-yl}pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid

[0160] tert-butyl (3R)-3-{6-bromo-lH-pyrrolo[2,3-b]pyridin-l-yl}pyrrolidine-l-carboxylate [0161] Prepared using a method similar to that employed in the first step of Example 1 using 6-bromo-lH-pyrrolo[2,3-b]pyridine (100 mg, 0.51 mmol) and tert-butyl (3S)-3- hydroxypyrrolidine-1 -carboxylate (2.25 equiv.). Yield: 166 mg, (87%) as a colorless oil. LCMS m/z found 310.0, MH⁺ -C₄H₈.

[0162] tert-butyl ( 3R)-3-{ 6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH.2H.3H- pyrctzolo[3, 4-d]pyrimidin-l-yl]-lH-pyrrolo[2,3-b]pyridin-l-yl}pyrrolidine-l-carboxylate [0163] Prepared using a method similar to that employed in the second step of Example 1 using tert-butyl (3R)-3-{6-bromo-lH-pyrrolo[2.3-b]pyridin-l-yl}pyrrolidine-l -carboxylate (166 mg) and trans- l,2-bis(methylamino)cy cl ohexane as the ligand. Yield: 231 mg (crude) as a pale-yellow solid. LCMS m/z found 408.0, MH⁺ -Boc.

[0164] 6-[( l-methylindazol-5-yl)amino ]-2-(prop-2-en- 1-yl)- !-{!-[ (3R)-pyrrolidin-3- yl ]pyrrolo[2, 3-b ]pyridin-6-yl}pyrazolo[3, 4-d]pyrimidin-3-one; trifluoroacetic acid

[0165] Prepared using a method similar to that employed in the third step of Example 1 using tert-butyl (3R)-3-{6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH,2H,3H-pyrazolo[3,4- d]pyrimidin-l-yl]-lH-pyrrolo[2,3-b]pyridin-l-yl}pyrrolidine-l-carboxylate (231 mg, crude) to give the free base intermediate (160 mg) as a brown solid of which 13% was purified by prep-HPLC. Yield: 11 mg TFA salt (30%) as a pale-yellow powder. LCMS m/z found 507.0, MH .

[0166] ‘ H NMR (400 MHz, dmso) 3 10.37 (s, 1H), 8.99 (d, J= 57.0 Hz, 2H), 8.90 (s, 1H), 8.42 - 8.24 (m, 2H), 7.99 (s, 1H), 7.79 (d, J= 3.2 Hz, 1H), 7.73 - 7.51 (m, 3H), 6.71 (d, J = 3.5 Hz, 1H), 5.80 - 5.64 (m, 1H), 5.57 - 5.41 (m, 1H), 5.03 (d, J= 10.2 Hz, 1H), 4.88 (d, J = 17.0 Hz, 1H), 4.64 (s, 2H). 4.02 (s, 3H). 3.77 - 3.69 (m, 1H), 3.58 - 3.50 (m. 1H), 3.48 - 3.35 (m, 2H), 2.57 - 2.53 (m, 1H), 2.37 - 2.27 (m, 1H). [0167] Example 6. Synthesis of 6-[(l-methylindazol-5-yl)amino]-l-{l-[(3R)-piperidin-3- yl] py rrolo [2,3-b] py ridin-6-yl }-2-(prop-2-en-l-yl)pyrazolo [3,4-d ] pyrimidin-3-one; trifluoroacetic acid (Compound 105).

[0168] tert-butyl (3R)-3-{6-bromopyrrolo[2,3-b]pyridm-l-yl}piperidine-l-carboxylate [0169] Prepared using a method similar to that employed in the first step of Example 1 using 6-bromo-lH-pyrrolo[2,3-b]pyridine (100 mg, 0.51 mmol) and tert-butyl (S)-3-hydroxy-l- piperidinecarboxylate (2.8 equiv.). Yield: 152 mg, (72%) as a colorless film. LCMS m/z found 324.0, MH⁺ -C₄H₈.

[0170] tert-butyl (3R)-3-{6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH,2H,3H- pyrazolo[3.4-d]pyrimidin-l-yl]-lH-pyrrolo[2,3-b]pyridin-l-yl}piperidine-l-carboxylate [0171] Prepared using a method similar to that employed in the second step of Example 1 using tert-butyl (3R)-3-{6-bromopyrrolo[2,3-b]pyridin-l-yl}piperidine-l-carboxylate (0.36 mmol) and N,N'-Dimethylethylenediamine as the ligand. Yield: 148 mg (78%) as a paleyellow oil. LCMS m/z found 522.0, MH⁺.

[0172] 6-[(l-methylindazol-5-yl)amino]-l-{l-[(3R)-piperidin-3-yl]pyrrolo[2,3-b]pyridin-6- y!}-2-(prop-2-en-l-yl)pyrazolo[3, 4-dJpyrimidin-3-one; trifluoroacetic acid

[0173] Prepared using a method similar to that employed in the third step of Example 1 using tert-butyl (3R)-3-{6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH,2H,3H-pyrazolo[3,4- d]pyrimidin-l-yl]-lH-pyrrolo[2,3-b]pyridin-l-yl}piperidine-l-carboxylate (148 mg) to give the free base intermediate (149 mg) as a brown solid of which 20% was purified by prep- HPLC. Yield: 17.4 mg TFA salt (48%) as a white powder. LCMS m/z Found: 521.0 MH⁺ [0174] 1H NMR (400 MHz, DMSO) 5 10.38 (s, 1H), 9.01 - 8.88 (m, 2H), 8.87 - 8.72 (m, 1H), 8.34 (s, 2H), 8.00 (s, 1H), 7.78 (d, J = 3.6 Hz, 1H). 7.68 (d, J = 5.7 Hz, 1H), 7.59 (s, 2H), 6.70 (d, J = 3.6 Hz, 1H), 5.73 (ddt, J = 16.5, 10.9, 5.9 Hz, 1H), 5.11 - 4.96 (m, 2H). 4.87 (d, J = 17.1 Hz, 1H), 4.67 (s, 2H), 4.02 (s, 3H), 3.46 - 3.27 (m, 3H), 2.96 - 2.82 (m, 1H), 2.23 - 2.09 (m, 1H), 2.08 - 1.96 (m, 2H), 1.86 (q, J = 11.9 Hz, 1H).

[0175] Example 7. Synthesis of 6-[(l-methylindazol-5-yl)amino]-l-{l-[(3S)-piperidin-3- yl] pyrrolo [2,3-b] py ridin-6-yl }-2-(prop-2-en-l-yl)pyrazolo [3,4-d] pyrimidin-3-one; trifluoroacetic acid (Compound 106)

[0176] tert-butyl (3S)-3-{6-bromo-lH-pyrrolo[2,3-b]pyridm-l-yl}piperidme-l-carboxylate [0177] Prepared using a method similar to that employed in the first step of Example 1 using 6-bromo-lH-pyrrolo[2,3-b]pyridine (100 mg, 0.51 mmol) and tert-butyl (3R)-3- hydroxypiperidine-1 -carboxylate (3 equiv.). Yield: 123 mg. (58%) as a colorless oil. LCMS m/z 324.0, MH⁺ -C₄H₈. [0178] tert-butyl (3S)-3-{ 6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH, 2H, 3H- pyrazolo[3.4-d ]pyrimidin- 1 -yl ]- lH-pyrrolo[2.3-b ]pyridin-l-yl}piperidine- 1 -carboxylate [0179] Prepared using a method similar to that employed in the second step of Example 1 using tert-butyl (3S)-3-{6-bromo-lH-pyrrolo[2,3-b]pyridin-l-yl}piperidine-l-carboxylate (123 mg) and trans- l,2-bis(methylamino)cy cl ohexane as the ligand. Yield: 204 mg (crude) as a pale-yellow solid. LCMS m/z found 522.0, MH⁺ .

[0180] 6-[(l-methylindazol-5-yl)amino]-l-{l-[(3S)-piperidin-3-yl]pyrrolo[2,3-b]pyridin-6- yl}-2-(prop-2-en-l-yl)pyrazolo[ 3, 4-d]pyrimidin-3-one; trifluoroacetic acid

[0181] Prepared using a method similar to that employed in the third step of Example 1 using tert-butyl (3S)-3-{6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH,2H,3H-pyrazolo[3,4- d]pyrimidin-l-yl]-lH-pyrrolo[2.3-b]pyridin-l-yl}piperidine-l-carboxylate (204 mg, crude) to give the free base intermediate (141 mg) as a brown solid of which 17% was purified by prep-HPLC. Yield: 11 mg TFA salt (26%) as a pale-yellow powder. LCMS m/z Found: 521.0 MH .

[0182] ¹H NMR (400 MHz, dmso) 5 10.39 (s. 1H), 9.01 - 8.74 (m, 3H), 8.34 (s, 2H), 8.00 (s, 1H), 7.78 (d. J= 3.3 Hz. 1H), 7.74 - 7.52 (m. 3H), 6.70 (d, J= 3.5 Hz. 1H), 5.81 - 5.64 (m, 1H), 5.11 - 4.99 (m, 2H), 4.87 (d, J= 16.1 Hz, 1H), 4.67 (s, 2H), 4.02 (s, 3H), 3.49 - 3.43 (m, 1H), 3.39 - 3.28 (m, 2H), 2.97 - 2.81 (m, 1H), 2.21 - 1.96 (m, 3H), 1.95 - 1.79 (m, 1H).

[0183] Example 8. Synthesis of l-{l-[(4S)-azepan-4-yl]-lH-pyrrolo[2,3-b]pyridin-6-yl}- 6-[(l-methyl-lH-indazol-5-yl)amino]-2-(prop-2-en-l-yl)-lH,2H,3H-pyrazolo[3,4- d]pyrimidin-3-one; trifluoroacetic acid (Compound 107).

[0184] tert-butyl ( 4S)-4-{ 6-bromo-lH-pyrrolo[2, 3-b ]pyridin-l-yl}azepane-l-carboxylate [0185] Prepared using a method similar to that employed in the first step of Example 1 using 6-bromo-lH-pyrrolo[2,3-b]pyridine (100 mg, 0.51 mmol) and tert-butyl (4R)-4- hydroxyazepane-1 -carboxylate (2 equiv.). Yield: 151 mg (75%) as a colorless oil. LCMS m/z 338.0, MH⁺ -C₄H₈.

[0186] tert-butyl (4S)-4-{6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH,2H,3H- pyrazolo[3.4-d]pyrimidin-l-yl]-lH-pyrrolo[2.3-b]pyridin-l-yl}azepane-l-carboxylate [0187] Prepared using a method similar to that employed in the second step of Example 1 using tert-butyl (4S)-4-{6-bromo-lH-pyrrolo[2,3-b]pyridin-l-yl}azepane-l-carboxylate (136 mg) and N,N'-Dimethylethylenediamine as the ligand. The crude material was used in the next step without purification. Yield: 160 mg (crude) as a yellow foam. [0188] l-{l-[(4S)-azepan-4-yl ]-lH-pyrrolo[2, 3-b ]pyridin-6-yl}-6-[ ( l-methyl-lH-indazol-5- yl)amino]-2-(prop-2-en-l-yl)-lH,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid

[0189] Prepared using a method similar to that employed in the third step of Example 1 using tert-butyl (4S)-4-{6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH,2H,3H-pyrazolo[3,4- d]pyrimidin-l-yl]-lH-pyrrolo[2,3-b]pyridin-l-yl}azepane-l-carboxylate (160 mg) to give the free base intermediate (158 mg) as a brown solid of which 20% was purified by prep-HPLC. Yield: 15.3 mg TFA salt (39%) as a yellow powder. LCMS m/z Found: 535.0, MH⁺.

[0190] 1H NMR (500 MHz, DMSO) 5 10.34 (s, 1H), 8.90 (s, 1H), 8.66 (s, 2H), 8.31 (s, 2H), 7.99 (s, 1H), 7.70 (d, J = 3.6 Hz, 1H), 7.65 (s, 1H), 7.63 - 7.59 (m, 1H), 7.58 (d, J = 9.0 Hz, 1H), 6.66 (d, J = 3.6 Hz, 1H), 5.73 (ddt, J = 16.3, 10.8, 5.7 Hz, 1H), 5.02 (dd, J = 10.3. 1.5 Hz, 1H), 4.96 (tt, J = 10.0, 5.1 Hz, 1H), 4.87 (d, J = 17.0 Hz, 1H), 4.66 (s, 2H), 4.03 (s, 3H), 3.41 - 3.33 (m, 1H), 3.32 - 3.23 (m, 2H), 3.22 - 3.11 (m, 1H), 2.41 - 2.30 (m, 1H), 2.26 - 2.06 (m, 3H), 2.03 - 1.82 (m, 2H).

[0191] Example 9. Synthesis of l-{l-[(4R)-azepan-4-yl]-lH-pyrrolo[2,3-b]pyridin-6-yl}- 6-[(l-methyl-lH-indazol-5-yl)amino]-2-(prop-2-en-l-yl)-lH,2H,3H-pyrazolo[3,4- d]pyrimidin-3-one; trifluoroacetic acid (Compound 108)

[0192] tert-butyl ( 4R)-4-{ 6-bromo-lH-pyrrolo[2, 3-b ]pyridin-l-yl}azepane-l-carboxylate [0193] Prepared using a method similar to that employed in the first step of Example 1 using 6-bromo-lH-pyrrolo[2,3-b]pyridine (100 mg, 0.51 mmol) and tert-butyl (4S)-4- hydroxyazepane-1 -carboxylate (2 equiv.). Yield: 153 mg (76%) as a colorless oil. LCMS m/z 338.0, MH⁺ -C₄H₈.

[0194] tert-butyl (4R)-4-{6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH,2H,3H- pyrazolo[3.4-d fpyrimidin- 1 -yl ]- lH-pyrrolo[2.3-b ]pyridin-l-yl fazepane- 1 -carboxylate [0195] Prepared using a method similar to that employed in the second step of Example 1 using tert-butyl (4R)-4-{6-bromo-lH-pyrrolo[2,3-b]pyridin-l-yl}azepane-l-carboxylate (157 mg) and N,N'-Dimethylethylenediamine as the ligand. The crude material was used in the next step without purification. Yield: 198 mg (crude) as a yellow foam.

[0196] l-{l-[(4R)-azepan-4-yl]-lH-pyrrolo[2,3-b]pyridin-6-yl}-6-[(l-methyl-lH-indazol-5- ylfamino ]-2-(prop-2-en-l-yl)-lH, 2H, 3H-pyr azolof 3, 4-d]pyrimidin-3-one; trifluoroacetic acid

[0197] Prepared using a method similar to that employed in the third step of Example 1 using tert-butyl (4R)-4-{6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH,2H,3H-pyrazolo[3,4- d]pyrimidin-l-yl]-lH-pyrrolo[2,3-b]pyridin-l-yl}azepane-l-carboxylate (198 mg) to give the free base intermediate (173 mg) as a brown solid of which 20% was purified by prep-HPLC. Yield: 14.6 mg TFA salt (30%) as a yellow powder. LCMS m/z Found: 535.0, MH⁺.

[0198] 1H NMR (400 MHz, dmso) 5 10.37 (s, 1H), 8.90 (s, 1H), 8.64 (s, 2H), 8.32 (s, 2H), 7.99 (s, 1H), 7.70 (d, J = 3.7 Hz, 1H), 7.68 - 7.53 (m, 3H), 6.65 (d, J = 3.6 Hz, 1H), 5.72 (ddt, J = 16.3, 10.9, 5.8 Hz, 1H), 5.01 (d, J = 10.3 Hz, 1H), 4.95 (dt, J = 10.5, 5.3 Hz, 1H), 4.85 (d, J = 17.2 Hz, 1H), 4.65 (s, 2H), 4.02 (s, 3H), 3.40 - 3.32 (m, 1H), 3.32 - 3.21 (m, 2H), 3.21 - 3.07 (m, 1H), 2.41 - 2.28 (m, 1H). 2.26 - 2.02 (m, 3H). 2.03 - 1.77 (m, 2H).

[0199] Example 10. Synthesis of 6-[(l-methyl-lH-indazol-5-yl)amino]-l-[3-(piperidin-4- yl)-3H-imidazo[4,5-b]pyridin-5-yl]-2-(prop-2-en-l-yl)-lH,2H,3H-pyrazolo[3,4- d]pyrimidin-3-one; trifluoroacetic acid (Compound 109)

[0200] tert-butyl 4-{5-bromo-3H-imidazo[4.5-b]pyridin-3-yl}piperidine-l -carboxylate

[0201] Prepared using a method similar to that employed in the first step of Example 1 using 5-bromo-3H-imidazo[4,5-b]pyridine (100 mg, 0.51 mmol) and tert-butyl 4- hydroxypiperidine-1 -carboxylate (1.5 equiv ). Yield: 157 mg (82%) as a brown oil, containing trace amount of PBu₃O. LCMS m/z found 381.0, MH⁺.

[0202] tert-butyl 4-{5-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH,2H,3H- pyrazolo[3, 4-d]pyrimidin-l-yl]-3H-imidazo[4.5-b]pyridin-3-yl}piperidine-l-carboxylate [0203] Prepared using a method similar to that employed in the second step of Example 1 using tert-butyl 4-{5-bromo-3H-imidazo[4,5-b]pyridin-3-yl}piperidine-l-carboxylate (157 mg) and trans- l,2-bis(methylamino)cyclohexane as the ligand. Yield: 226 mg (crude) as a brown solid. LCMS m/z found 423.0, MH⁺ -Boc.

[0204] 6-[(l-methyl-lH-indazol-5-yl)amino]-l-[3-(piperidin-4-yl)-3H-imidazo[4,5- b]pyridin-5-yl]-2-(prop-2-en-l-yl)-lH,2H3H-pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid

[0205] Prepared using a method similar to that employed in the third step of Example 1 using tert-butyl 4-{5-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH,2H,3H-pyrazolo[3,4- d]pyrimidin-l-yl]-3H-imidazo[4,5-b]pyridin-3-yl}piperidine-l-carboxylate (215 mg, crude) to give the free base intermediate (185 mg) as a brown solid of which 12% was purified by prep-HPLC. Yield: 15 mg TFA salt (48%) as a pale-yellow powder. LCMS m/z Found: 522.0, MH⁺.

[0206] ^JH NMR (400 MHz, dmso) 5 10.40 (s, 1H), 8.91 (s, 1H), 8.75 (d, J= 8.9 Hz, 1H), 8.60 (s, 1H), 8.56 - 8.39 (m, 2H), 8.25 (s, 1H), 8.03 (s, 1H), 7.85 (d, J= 8.0 Hz, 1H), 7.69 - 7.54 (m, 2H), 5.84 - 5.58 (m, 1H). 5.00 (d, J= 10.2 Hz. 1H), 4.92 - 4.78 (m. 2H), 4.67 (s, 2H), 4.03 (s, 3H), 3.47 (d, J= 11.5 Hz, 2H), 3.19 (q, J= 10.5 Hz, 2H), 2.36 - 2.20 (m, 4H). [0207] Example 11. Synthesis of 6-[(l-methyl-lH-indazol-5-yl)amino]-2-(prop-2-en-l- yl)-l-{3- [(3S)-pyrrolidin-3-yl] -3H-imidazo [4,5-b] pyridin-5-yl}-lH,2H,3H-pyrazolo [3,4- d]pyrimidin-3-one; trifluoroacetic acid (Compound 110)

[0208] tert-butyl (3S)-3-{5-bromo-3H-imidazo[4,5-b]pyridin-3-yl}pyrrolidme-l-carboxylate [0209] Prepared using a method similar to that employed in the first step of Example 1 using 5-bromo-3H-imidazo[4,5-b]pyridine (100 mg. 0.51 mmol) and tert-butyl (3R)-3- hydroxypyrrolidine-1 -carboxylate (1.5 equiv.). Traces of PBmO remaining after chromatography were removed by washing with diethyl ether followed by drying in vacuo. The connectivity was confirmed by HMBC observation of the coupling between C3 -H-

C3a. Yield: 67 mg (36%), as a white powder LCMS m/z found 367.0, MH⁺.

[0210] tert-butyl (3S)-3-{5-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH.2H,3H- pyrazolo[3, 4-d]pyrimidin-l-yl]-3H-imidazo[4.5-b]pyridin-3-yl}pyrrolidine-l-carboxylate [0211] Prepared using a method similar to that employed in the second step of Example 1 using tert-butyl (3S)-3-{5-bromo-3H-imidazo[4,5-b]pyridin-3-yl}pyrrolidine-l-carboxylate (64 mg) and trans- l,2-bis(methylamino)cy cl ohexane as the ligand. Yield: 102 mg (crude) as a brown solid. LCMS m/z found 409.0, MH⁺ -Boc.

[0212] 6-[(l -methyl-lH-indazol-5-yl)amino]-2-(prop-2-en-l-yl)-l-{3-[(3S)-pyrrolidin-3-yl]- 3H-imidazo[4, 5-b ]pyridin-5-yl}-lH, 2H, 3H-pyrazolo[3, 4-d]pyrimidin- 3-one; trifluoroacetic acid

[0213] Prepared using a method similar to that employed in the third step of Example 1 using tert-butyl (3S)-3-{5-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l -yl)-lH,2H,3H-pyrazolo[3,4- d]pyrimidin-l-yl]-3H-imidazo[4,5-b]pyridin-3-yl}pyrrolidine-l-carboxylate (89 mg, crude) to give the free base intermediate (109 mg) as a brown solid of which 19% was purified by prep-HPLC. Yield: 8 mg TFA salt (39%) as a pale-yellow powder. LCMS m/z Found: 508.0, MH .

[0214] ¹H NMR (400 MHz, dmso) 8 10.43 (s, 1H), 9.02 (d, J= 26.3 Hz, 2H), 8.91 (s, 1H), 8.66 (s, 1H), 8.50 (s, 1H), 8.26 (s, 1H), 8.03 (s, 1H), 7.86 (d, J= 8.1 Hz, 1H), 7.69 - 7.55 (m, 2H), 5.78 - 5.63 (m, 1H), 5.37 (p, J= 6.3 Hz, 1H), 5.03 (d, J= 9.9 Hz, 1H), 4.89 (d, J= 17.2 Hz. 1H), 4.63 (s. 2H), 4.03 (s, 3H), 3.80 - 3.72 (m, 1H), 3.68 - 3.55 (m, 2H). 3.47 - 3.39 (m, 1H), 2.61 - 2.53 (m, 2H).

[0215] Example 12. Synthesis of 6-[(l-methyl-lH-indazol-5-yl)amino]-l-{3-[(3S)- piperidin-3-yl]-3H-imidazo[4,5-b]pyridin-5-yl}-2-(prop-2-en-l-yl)-lH,2H,3H- pyrazolo[3,4-d]pyrimidin-3-one (Compound 111)

[0216] tert-butyl (3S)-3-{5-bromo-3H-imidazo[4,5-b]pyridin-3-yl}piperidine-l-carboxylate [0217] Prepared using a method similar to that employed in the first step of Example 1 using

5-bromo-3H-imidazo[4,5-b]pyridine (100 mg. 0.51 mmol) and tert-butyl (3R)-3- hydroxypiperidine-1 -carboxylate (2.5 equiv.). Yield: 47 mg (24%) as a brown oil, containing trace amount of PBmO LCMS m/z found 381.0, MH⁺.

[0218] tert-butyl (3S)-3-{5-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH, 2H, 3H- pyrazolo[3.4-d]pyrimidin-l-yl]-3H-imidazo[4,5-b]pyridin-3-yl}piperidine-l-carboxylale [0219] Prepared using a method similar to that employed in the second step of Example 1 using tert-butyl (3S)-3-{5-bromo-3H-imidazo[4,5-b]pyridin-3-yl}piperidine-l -carboxylate (47 mg) and trans-l,2-bis(methylamino)cyclohexane as the ligand. Yield: 74 mg (crude) as a brown solid. LCMS m/z found 523.0, MH⁺.

[0220] 6-[( I -methyl- lH-indazol-5-yl)amino ]-!-{ 3-[ ( 3S)-piperidin-3-yl ]-3H-imidcizo[4, 5- b]pyridin-5-yl}-2-(prop-2-en-l-yl)-lH.2H.3H-pyrazolo[3, 4-d]pyrimidin-3-one; trifluoroacetic acid

[0221] Prepared using a method similar to that employed in the third step of Example 1 using tert-butyl (3S)-3-{5-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH,2H,3H-pyrazolo[3,4- d]pyrimidin-l-yl]-3H-imidazo[4,5-b]pyridin-3-yl}piperidine-l-carboxylate (79 mg, crude) to give the free base intermediate (83 mg) as a brown solid of which 21% was purified by prep- HPLC. Yield: 5 mg TFA salt (30%) as a pale-yellow powder. LCMS m/z Found: 522.0 Calc: 522.3 for C27H27N11O I H .

[0222] ¹H NMR (400 MHz, dmso) 6 10.43 (s. 1H), 8.96 (d. J= 9.1 Hz. 1H), 8.91 (s, 1H), 8.83 (q, .7= 9.1 Hz, 1H), 8.68 (s, 1H), 8.50 (s, 1H), 8.26 (s, 1H), 8.03 (s, 1H), 7.86 (d, J= 8.4 Hz, 1H), 7.67 - 7.56 (m, 2H), 5.78 - 5.62 (m, 1H), 5.02 (d, J= 10.2 Hz, 1H), 4.95 - 4.84 (m, 2H), 4.66 (s, 2H), 4.03 (s, 3H), 3.61 (d, J= 11.9 Hz, 2H), 3.43 - 3.34 (m, 1H), 2.89 (q, J = 11.7 Hz, 1H), 2.29 - 2.13 (m, 2H), 2.03 (d, J= 13.1 Hz, 1H), 1.85 (q. J= 12.8 Hz, 1H). Overlap with water peak.

[0223] Example 13. Synthesis of 6-[(l-methyl-lH-indazol-5-yl)amino]-2-(prop-2-en-l- yl)-l-{l-[(3S)-pyrrolidin-3-yl]-lH-pyrazolo[3,4-b]pyridin-6-yl}-lH,2H,3H-pyrazolo[3,4- d]pyrimidin-3-one; trifluoroacetic acid (Compound 112)

[0224] tert-butyl ( 3S)-3-{ 6-bromo-lH-pyrazolo[3.4-b ]pyridin- l-yl)pyrrolidine- 1 -carboxylate [0225] Prepared using a method similar to that employed in the first step of Example 1 using

6-bromo-lH-pyrazolo[3,4-b]pyridine (100 mg, 0.51 mmol) and tert-butyl (3R)-3- hydroxypyrrolidine-1 -carboxylate (2.5 equiv.). Yield: 81 mg (44%) as a yellow oil. LCMS m/z found 310.8. MH⁺ -C4H8. [0226] tert-butyl (3S)-3-{ 6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH, 2H, 3H- pyrazolo[3.4-d]pyrimidin- 1 -yl]- lH-pyrazolo[3,4-b]pyridin- 1 -yl}pyrrolidine- 1-carboxylate [0227] Prepared using a method similar to that employed in the second step of Example 1 using tert-butyl (3S)-3- {6-bromo-lH-pyrazolo[3,4-b]pyridin-l -yl} pyrrolidine-1 -carboxylate (81 mg) and trans- l,2-bis(methylamino)cy cl ohexane as the ligand. Yield: 117 mg (crude) as a brown solid. LCMS m/z found 409.0, MH⁺ -Boc.

[0228] 6-[(l-methyl-lH-indazol-5-yl)amino]-2-(prop-2-en-l-yl)-l-{l-[(3S)-pyrrolidin-3-yl]- lH-pyrazolo[3, 4-b ]pyridin-6-yl}-lH, 2H, 3H-pyrazolo[ 3, 4-d]pyrimidin-3-one; trifluoroacetic acid

[0229] Prepared using a method similar to that employed in the third step of Example 1 using tert-butyl (3S)-3-{6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH,2H,3H-pyrazolo[3,4- d]pyrimidin-l-yl]-lH-pyrazolo[3,4-b]pyridin-l-yl}pyrrolidine-l-carboxylate (117 mg, crude) to give the free base intermediate (107 mg) as a brow n solid of which 19% w as purified by prep-HPLC. Yield: 9.5 mg TFA salt (35%) as a pink powder. LCMS m/z Found: 508.0, MH⁺. [0230] ¹H NMR (400 MHz, dmso) 5 10.52 (s. 1H), 9.18 (s, 1H), 9.05 (s, 1H), 8.94 (s, 1H). 8.57 (d, J= 7.1 Hz, 1H). 8.37 - 8.25 (m. 2H). 8.08 (s. 1H), 7.94 (d. J= 8.4 Hz. 1H), 7.62 (s, 2H), 5.80 - 5.64 (m, 2H), 5.02 (d, J= 10.2 Hz, 1H), 4.90 (d, J= 17.4 Hz, 1H), 4.77 (d, J = 5.7 Hz, 2H), 4.05 (s, 3H), 3.74 (dt, J= 10.7, 5.4 Hz, 1H), 3.66 - 3.58 (m, 2H), 3.49 (dd, J = 14.2, 6.2 Hz, 2H), 2.41 - 2.28 (m, 1H).

[0231] Example 14. Synthesis of 6-[(l-methyl-lH-indazol-5-yl)amino]-2-(prop-2-en-l- yl)-l -{1 - [(3R)-pyrrolidin-3-yl] -1 H-pyrazolo [3,4-b] pyridin-6-yl}-l H,2H,3H-pyrazolo [3,4- d]pyrimidin-3-one; trifluoroacetic acid (Compound 113)

[0232] tert-butyl (3R)-3-{6-bromo-lH-pyrazolo[3,4-b]pyridm-l-yl}pyrrolidine- 1-carboxylate [0233] Prepared using a method similar to that employed in the first step of Example 1 using 6-bromo-lH-pyrazolo[3,4-b]pyridine (100 mg, 0.51 mmol) and tert-butyl (3S)-3- hydroxypyrrolidine- 1-carboxylate (2.5 equiv.). Yield: 83 mg (45%) as a yellow oil.

[0234] tert-butyl (3R)-3-{6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH,2H,3H- pyrazolo[3.4-d]pyrimidin-l-yl]-lH-pyrazolo[3,4-b]pyridin-l-yl}pyrrolidine-l-carboxylate [0235] Prepared using a method similar to that employed in the second step of Example 1 using tert-butyl (3R)-3-{6-bromo-lH-pyrazolo[3,4-b]pyridin-l-yl}pyrrolidine-l -carboxylate (83 mg) and trans-l,2-bis(methylamino)cyclohexane as the ligand. Yield: 130 mg (crude) as a brown solid. LCMS m/z found 409.0, MH⁺ -Boc. [0236] 6-[( 1 -methyl- lH-indazol-5-yl)amino ]-2-(prop-2-en-l-yl)-l-{ l-[(3R)-pyrrolidin-3-yl ]- lH-pyrazolo[3.4-b ]pyridin-6-yl}-lH, 2H, 3H-pyrazolo[ 3, 4-d]pyrimidin-3-one; trifluoroacetic acid

[0237] Prepared using a method similar to that employed in the third step of Example 1 using tert-butyl (3R)-3-{6-[6-(methylsulfanyl)-3-oxo-2-(prop-2-en-l-yl)-lH,2H,3H-pyrazolo[3,4- d]pyrimidin-l-yl]-lH-pyrazolo[3,4-b]pyridin-l-yl}pyrrolidine-l-carboxylate (130 mg, crude) to give the free base intermediate (104 mg) as a brown solid of which 18% was purified by prep-HPLC. Yield: 7.1 mg TFA salt (25%) as a pink powder. LCMS m/z Found: 508.0, MH⁺. [0238] ¹H NMR (400 MHz, dmso) 5 10.51 (s, 1H), 9.16 (s, 1H), 9.03 (s, 1H), 8.94 (s, 1H), 8.57 (d, J= 7.4 Hz, 1H), 8.36 - 8.24 (m. 2H), 8.08 (s, 1H), 7.94 (d, J= 8.8 Hz, 1H), 7.62 (s, 2H), 5.79 - 5.64 (m, 2H), 5.02 (d, J= 10.1 Hz, 1H). 4.90 (d, J= 17.2 Hz. 1H), 4.77 (d. J= 6.0 Hz, 2H), 4.05 (s, 3H), 3.73 (dt, J= 12.4, 6.4 Hz, 1H), 3.67 - 3.58 (m, 2H), 3.47 (dd, J = 14.1, 6.6 Hz, 2H), 2.35 (dd, J= 12.6, 7.5 Hz, 1H).

[0239] Example 15. Synthesis of 6-[(l-Methylindazol-5-yl)amino]-l-{l-[(3S)-l- methylpyrrolidin-3-yl] pyrrolo [2,3-b] pyridin-6-yl }-2-(prop-2-en-l-yl)pyrazolo [3,4- d]pyrimidin-3-one; trifluoroacetic acid (Compound 114)

[0240] Aqueous formaldehyde (37%, 0.72 mmol, 2.0 equiv.) followed by sodium triacetoxyborohydride (153 mg, 0.72 mmol, 2.0 equiv.) were added to a solution of 6-[(l - methylindazol-5-yl)amino]-2-(prop-2-en-l-yl)-l-{l-[(3S)-pyrrolidin-3-yl]pyrrolo[2,3- b]pyridin-6-yl}pyrazolo[3.4-d]pyrimidin-3-one (183 mg, 0.36 mmol. 1.0 equiv.) in THF (12 ml) at RT and the resulting mixture was stirred until the secondary amine was consumed (18 h, LCMS). The mixture was partitioned between EtOAc and sat. brine and pH adjusted to ca. 11 with aq. NaOH. The organic phase was filtered through a phase-separator and concentrated under reduced pressure. The residue was dissolved in MeOH/H2O and purified by reversed phase chromatography (Gemini NX-C18, 21x150 mm, water (0.1% TFA)/acetonitrile, gradient over 12 minutes, 25 ml/min) and the pure fractions were lyophilized to give the title compound (TFA salt, 130 mg, 57%). LCMS m/z Found: 521.0, MH .

[0241] ¹H NMR (400 MHz, dmso) 6 10.55 - 9.82 (m. 2H), 8.90 (s. 1H), 8.45 - 8.22 (m, 2H), 8.00 (d, J= 5.2 Hz, 1H), 7.80 (d, J= 33.8 Hz, 1H), 7.74 - 7.54 (m, 3H), 6.72 (d, J= 3.3 Hz, 1H), 5.78 - 5.44 (m, 2H), 5.04 (d, J= 10.1 Hz, 1H), 4.89 (d, J= 18.0 Hz, 1H), 4.64 (s, 2H), 4.02 (s, 3H), 3.90 - 3.62 (m, 2H), 3.57 - 3.21 (m, 2H), 2.94 (dd, J= 12.3, 4.3 Hz, 3H), 2.79 - 2.69 (m, 1H), 2.34 - 2.23 (m, 1H). [0242] Example 16. Synthesis of 6-[(l-methylindazol-5-yl)amino]-l-[l-(l- methylpiperidin-4-yl)pyrrolo[2,3-b]pyridin-6-yl]-2-(prop-2-en-l-yl)pyrazolo[3,4- d]pyrimidin-3-one (Compound 115).

[0243] Prepared using a method similar to that employed in Example 15 using 6-[(l - methylindazol-5-yl)amino]-l-[l-(piperidin-4-yl)pyrrolo[2,3-b]pyridin-6-yl]-2-(prop-2-en-l- yl)pyrazolo[3,4-d]pyrimidin-3-one (115 mg, crude). After purification by reversed-phase chromatography the pure fractions were loaded on an SCX-2 cartridge (1 g, 0.67 mEq/g), washed with MeOH, eluted with NEE/MeOH (1.4 M) and concentrated under reduced pressure. Yield: 57.5 mg free base (49%) as a white powder. LCMS m/z Found: 535.0 MH⁺. [0244] ¹H NMR (400 MHz, DMSO) 6 10.33 (s, 1H), 8.89 (s, 1H), 8.29 (s, 2H), 7.98 (s, 1H), 7.75 (d, J = 3.6 Hz, 1H), 7.59 (q. J = 9.3 Hz. 3H), 6.61 (d. J = 3.6 Hz. 1H), 5.74 (ddt. J = 16.4, 10.8, 5.8 Hz, 1H), 5.01 (d, J = 10.3 Hz, 1H), 4.87 (d, J = 17.2 Hz, 1H), 4.60 (d, J = 23.3 Hz, 3H), 4.02 (s, 3H), 2.89 (d, J = 6.8 Hz, 2H), 2.22 (s, 3H), 2.10 (s, 4H), 1.87 (s, 2H).

[0245] Example 17. Synthesis of 6-[(l-methylindazol-5-yl)amino]-l-[l-(l- methylpiperidin-4-yl)pyrazolo[3,4-b]pyridin-6-yl]-2-(prop-2-en-l-yl)pyrazolo[3,4- d]pyrimidin-3-one (Compound 116)

[0246] Prepared using a method similar to that employed in Example 15 using 6-[(l- methylindazol-5-yl)amino]-l-[l-(piperidin-4-yl)pyrazolo[3,4-b]pyridin-6-yl]-2-(prop-2-en-l- yl)pyrazolo[3,4-d]pyrimidin-3-one (98 mg, crude). The title compound was converted into the free base by solid-phase extraction (1 g SCX-2, MeOH. NHs/MeOH 1.4 M). Yield: 58.6 mg free base (58%) as a white powder. LCMS m/z Found: 536.0, MH⁺.

[0247] ¹H NMR (400 MHz, dmso) 5 10.47 (s, 1H), 8.92 (s, 1H), 8.51 (d, J = 8.6 Hz, 1H), 8.27 (s, 1H), 8.22 (s, 1H), 8.07 (s, 1H), 7.88 (d, J = 8.6 Hz, 1H), 7.62 (s, 2H), 5.75 (ddt, J = 16.4, 10.8, 5.8 Hz, 1H), 5.01 (d, J = 10.3 Hz, 1H). 4.87 (d, J = 17.2 Hz, 1H), 4.73 (d, J = 6.2 Hz, 3H), 4.04 (s, 3H), 2.91 (d, J = 10.2 Hz, 2H), 2.23 (s, 4H), 2.20 - 2.09 (m, 3H), 1.90 (d, J = 11.7 Hz, 2H).

[0248] Example 18. Synthesis of 6-[(l-methylindazol-5-yl)amino]-l-{l-[(3R)-l- methylpyrrolidin-3-yl]pyrrolo[2,3-b]pyridin-6-yl}-2-(prop-2-en-l-yl)pyrazolo[3,4- d]pyrimidin-3-one; trifluoroacetic acid (Compound 117)

[0249] Prepared using a method similar to that employed in Example 15 using 6-[( 1 - methylindazol-5-yl)amino]-2-(prop-2-en-l-yl)-l-{l-[(3R)-pyrrolidin-3-yl]pyrrolo[2,3- b]pyridin-6-yl}pyrazolo[3.4-d]pyrimidin-3-one (139 mg, crude). Yield: 84 mg TFA salt (48%) as a pale-yellow powder. LCMS m/z m/z Found: 521.0, MH⁺. [0250] NMR (400 MHz, dmso) 5 10.53 - 9.79 (m, 2H), 8.90 (s, 1H), 8.46 - 8.21 (m, 2H), 8.00 (d, J= 5.3 Hz, 1H), 7.80 (d, J= 34.5 Hz, 1H), 7.68 (s, 1H), 7.59 (s, 2H), 6.72 (s, 1H). 5.79 - 5.46 (m, 2H), 5.04 (d, J= 10.2 Hz, 1H), 4.89 (d, J = 17.1 Hz, 1H), 4.63 (s, 2H), 4.02 (s, 3H), 3.91 - 3.79 (m, 1H), 3.78 - 3.61 (m, 1H), 3.56 - 3.37 (m, 1H), 3.28 (p, J= 9.4 Hz, 1H), 2.94 (dd, J= 12.2, 4.0 Hz, 3H), 2.80 - 2.69 (m, 1H), 2.35 - 2.23 (m, 1H).

[0251] Example 19. Synthesis of 6-[(l-methylindazol-5-yl)amino]-l-{l-[(3R)-l- methylpiperidin-3-yl]pyrrolo[2,3-b]pyridin-6-yl}-2-(prop-2-en-l-yl)pyrazolo[3,4- d]pyrimidin-3-one; trifluoroacetic acid (Compound 118)

[0252] Prepared using a method similar to that employed in Example 15 using 6-[(l - methylindazol-5-yl)amino]-l-{l-[(3R)-piperidin-3-yl]pyrrolo[2,3-b]pyridin-6-yl}-2-(prop-2- en-l-yl)pyrazolo[3,4-d]pyrimidin-3-one (118 mg. crude). Yield: 73.8 mg TFA salt (50%) as a white powder. LCMS m/z Found: 535.0, MH⁺.

[0253] ¹H NMR (400 MHz, DMSO) 8 10.39 (s, 1H), 9.92 (s, 1H), 8.90 (s, 1H), 8.35 (s, 2H), 8.00 (s, 1H), 7.72 (d, J = 3.7 Hz, 1H), 7.71 - 7.65 (m, 1H), 7.59 (s, 2H), 6.73 (d, J = 3.6 Hz, 1H), 5.73 (ddt, J = 16.2, 10.6, 5.6 Hz, 1H), 5.07 (s, 1H). 5.02 (d, J = 10.3 Hz. 1H), 4.87 (d, J = 17.1 Hz. 1H), 4.67 (s, 2H). 4.02 (s, 3H). 3.55 (dd. J = 23.0, 11.8 Hz, 2H). 3.36 (q, J = 10.9 Hz, 1H), 2.95 (q, J = 1 1.6 Hz, 1H), 2.81 (d, J = 4.2 Hz, 3H), 2.13 - 2.00 (m, 3H), 1.98 - 1.78 (m, 1H).

[0254] Example 20. Synthesis of 6-[(l-methylindazol-5-yl)amino]-l-{l-[(3S)-l- methylpiperidin-3-yl]pyrrolo[2,3-b]pyridin-6-yl}-2-(prop-2-en-l-yl)pyrazolo[3,4- d]pyrimidin-3-one; trifluoroacetic acid (Compound 119)

[0255] Prepared using a method similar to that employed in Example 15 using 6-[(l - methylindazol-5-yl)amino]-l-{l-[(3S)-piperidin-3-yl]pyrrolo[2,3-b]pyridin-6-yl}-2-(prop-2- en-l-yl)pyrazolo[3,4-d]pyrimidin-3-one (117 mg, crude). Yield: 61 mg TFA salt (42%) as a pale-yellow powder. LCMS m/z Found: 535.0, MH⁺.

[0256] ‘H NMR (400 MHz, dmso) 3 10.39 (s, 1H), 9.94 (s, 1H), 8.90 (s, 1H), 8.45 - 8.22 (m, 2H), 8.01 (s, 1H), 7.79 - 7.53 (m, 4H), 6.73 (d, J= 3.5 Hz, 1H), 5.81 - 5.66 (m, 1H), 5.14 - 4.98 (m, 2H), 4.87 (d, J= 17.0 Hz, 1H), 4.68 (s. 2H), 4.02 (s, 3H), 3.55 (dd, J= 22.1, 11.7 Hz. 2H), 3.36 (q. J= 9.9 Hz. 1H), 3.03 - 2.89 (m. 1H), 2.81 (s, 3H), 2.12 - 2.00 (m, 3H), 1.97 — 1.81 (m, 1H).

[0257] Example 21. Synthesis of 6-[(l-methyl-lH-indazol-5-yl)amino]-l-[3-(l- methylpiperidin-4-yl)-3H-imidazo[4,5-b]pyridin-5-yl]-2-(prop-2-en-l-yl)-lH,2H,3H- pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid (Compound 120) [0258] Prepared using a method similar to that employed in Example 15 using 6-[(l-methyl- lH-indazol-5-yl)amino]-l-[3-(piperidin-4-yl)-3H-imidazo[4,5-b]pyridin-5-yl]-2-(prop-2-en-

1-yl)-lH,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (162 mg, crude). Yield: 1 10 mg TFA salt (55%) as a pale-yellow powder. LCMS m/z Found: 536.0, MH⁺.

[0259] ¹H NMR (400 MHz, dmso) 5 10.44 (s, 1H), 10.06 (d, J = 36.9 Hz, 1H), 8.91 (s, 1H), 8.68 (d, J= 13.4 Hz, 1H), 8.51 (s, 1H), 8.26 (s, 1H), 8.04 (s, 1H), 7.93 - 7.75 (m, 1H), 7.70 - 7.54 (m, 2H), 5.79 - 5.64 (m, 1H). 5.61 - 5.35 (m, 1H). 5.04 (d, J= 9.3 Hz, 1H). 4.90 (d, J = 17.5 Hz, 1H), 4.63 (s, 2H), 4.20 - 3.87 (m, 5H), 3.73 (d, J= 30.3 Hz, 1H), 3.61 - 3.25 (m, 1H), 2.95 (d, J= 16.9 Hz, 3H), 2.85 - 2.54 (m, 2H).

[0260] Example 22. Synthesis of 6-[(l-methyl-lH-indazol-5-yl)amino]-l-{3-[(3S)-l- methylpyrrolidin-3-yl]-3H-imidazo[4,5-b]pyridin-5-yl}-2-(prop-2-en-l-yl)-lH,2H,3H- pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid (Compound 121)

[0261] Prepared using a method similar to that employed in Example 15 using 6-[(l-methyl- lH-indazol-5-yl)amino]-2-(prop-2-en-l-yl)-l-{3-[(3S)-pyrrolidin-3-yl]-3H-imidazo[4,5- b]pyridin-5-yl}-lH,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (88 mg, crude). Yield: 50 mg TFA salt (45%) as a pale-yellow powder. LCMS m/z Found: 522.0, MH⁺.

[0262] ¹H NMR (400 MHz, dmso) 8 10.44 (s, 1H), 10.06 (d, J = 36.9 Hz, 1H), 8.91 (s, 1H), 8.68 (d, J= 13.4 Hz, 1H), 8.51 (s, 1H), 8.26 (s, 1H), 8.04 (s, 1H), 7.93 - 7.75 (m, 1H), 7.70 - 7.54 (m, 2H), 5.79 - 5.64 (m, 1H). 5.61 - 5.35 (m, 1H), 5.04 (d, J= 9.3 Hz, 1H), 4.90 (d, J = 17.5 Hz, 1H), 4.63 (s, 2H). 4.20 - 3.87 (m, 5H). 3.73 (d, J= 30.3 Hz. 1H), 3.61 - 3.25 (m. 1H), 2.95 (d, J = 16.9 Hz, 3H), 2.85 - 2.54 (m, 2H).

[0263] Example 23. Synthesis of 6-[(l-methyl-lH-indazol-5-yl)amino]-l-{3-[(3S)-l- methylpiperidin-3-yl]-3H-imidazo[4,5-b]pyridin-5-yl}-2-(prop-2-en-l-yl)-lH,2H,3H- pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid (Compound 122)

[0264] Prepared using a method similar to that employed in Example 15 using 6-[(l-methyl- 1 H-indazol-5-yl)amino] - 1 - { 3- [(3 S)-piperi din-3 -y 1] -3H-imidazo [4,5 -b] pyridin-5-yl } -2-(prop-

2-en-l-yl)-lH,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (66 mg). Yield: 24.5 mg TFA salt (39%) as a pale-yellow powder. LCMS m/z Found: 536.0, MH⁺.

[0265] ¹H NMR (400 MHz, dmso) 6 10.43 (s. 1H), 9.95 (s. 1H), 8.91 (s, 1H), 8.66 (s, 1H). 8.52 (s, 1H), 8.26 (s, 1H), 8.04 (s, 1H), 7.87 (d, J= 8.1 Hz, 1H), 7.70 - 7.52 (m, 2H), 5.80 - 5.64 (m, 1H), 5.02 (d, J= 10.2 Hz, 1H), 4.96 - 4.84 (m, 2H), 4.67 (s, 2H), 4.03 (s, 3H), 3.71 (d, J= 9.4 Hz, 1H), 3.55 (d, J= 11.4 Hz, 1H), 3.45 - 3.34 (m, 1H), 3.03 - 2.87 (m, 1H), 2.83 (s, 3H), 2.24 - 2.17 (m, 2H). 2.11 (d, J= 14.1 Hz. 1H), 1.94 - 1.78 (m, 1H). Overlap with water peak. [0266] Example 24. Synthesis of 6-[(l-methyl-lH-indazol-5-yl)amino]-l-{l-[(3S)-l- methylpiperidin-3-yl]-lH-pyrazolo[3,4-b]pyridin-6-yl}-2-(prop-2-en-l-yl)-lH,2H,3H- pyrazolo[3,4-d]pyrimidin-3-one (Compound 123)

[0267] Prepared using a method similar to that employed in Example 15 using 6-[(l-methyl- lH-indazol-5-yl)amino]-l-{l-[(3S)-piperidin-3-yl]-lH-pyrazolo[3,4-b]pyridin-6-yl}-2-(prop- 2-en-l-yl)-lH,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (45 mg). After purification by reversed-phase chromatography the pure fractions were loaded on an SCX-2 cartridge (1 g, 0.67 rnEq/g), washed with MeOH, eluted with and concentrated under reduced pressure. Yield: 11.5 mg free base (25%) as a powder. LCMS m/z Found: 536.0, MH .

[0268] ¹H NMR (400 MHz, dmso) 5 10.48 (s. 1H), 8.93 (s, 1H), 8.52 (d, J = 8.6 Hz, 1H), 8.28 (s, 1H), 8.22 (s, 1H), 8.07 (s, 1H), 7.89 (d, J = 8.6 Hz, 1H), 7.62 (s, 2H), 5.74 (ddt, J = 16.5, 10.9, 5.8 Hz, 1H), 5.01 (d, J = 10.2 Hz, 1H), 4.96 - 4.81 (m, 2H), 4.74 (d, J = 6.0 Hz, 2H), 4.04 (s, 3H), 3.01 - 2.90 (m, 1H), 2.81 (d, J = 11.3 Hz, 1H), 2.33 (t, J = 10.7 Hz, 1H),

2.21 (s, 3H), 2.06 - 1.87 (m, 3H), 1.87 - 1.61 (m, 2H).

[0269] Example 25. Synthesis of6-[(l-methyl-lH-indazol-5-yl)amino]-l-{l-[(4S)-l- methylazepan-4-yl]-lH-pyrrolo[2,3-b]pyridin-6-yI}-2-(prop-2-en-l-yl)-lH,2H,3H- pyrazolo[3,4-d]pyrimidin-3-one (Compound 124)

[0270] Prepared using a method similar to that employed in Example 15 using 1-{1-[(4S)- azepan-4-yl]-lH-pyrrolo[2,3-b]pyridin-6-yl}-6-[(l-methyl-lH-indazol-5-yl)amino]-2-(prop- 2-en-l-yl)-lH,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (126 mg). After purification by reversed-phase chromatography the pure fractions were loaded on an SCX-2 cartridge (1 g, 0.67 mEq/g), washed with MeOH, eluted with ) and concentrated under reduced pressure. Yield: 45 mg free base (35%) as a brown solid. LCMS m/z Found: 549.0, MH .

[0271] 1H NMR (400 MHz, dmso) 5 10.34 (s, 1H), 8.89 (s, 1H), 8.28 (s, 2H), 7.98 (s, 1H), 7.74 (d, J = 3.6 Hz, 1H), 7.61 (d, J = 8.8 Hz, 2H), 7.57 (d, J = 8.9 Hz, 1H), 6.59 (d, J = 3.5 Hz. 1H), 5.73 (ddt. J = 16.2, 10.7, 5.7 Hz, 1H), 5.00 (d, J = 10.2 Hz, 1H). 4.96 - 4.89 (m, 1H), 4.85 (d. J = 17.3 Hz, 1H). 4.64 (s, 2H). 4.02 (s. 3H), 2.71 - 2.54 (m. 4H), 2.29 (s, 3H),

2.22 - 2.03 (m, 2H), 2.03 - 1.89 (m, 2H), 1.87 - 1.75 (m, 1H), 1.75 - 1.62 (m, 1H).

[0272] Example 26. Synthesis of 6-[(l-methyl-lH-indazol-5-yl)amino]-l-{l-[(4R)-l- methylazepan-4-yl]-lH-pyrrolo[2,3-b]pyridin-6-yl}-2-(prop-2-en-l-yl)-lH,2H,3H- pyrazolo[3,4-d]pyrimidin-3-one (Compound 125) [0273] Prepared using a method similar to that employed in Example 15 using 1- { 1 -[(4R)- azepan-4-yl]-lH-pyrrolo[2,3-b]pyridin-6-yl}-6-[(l-methyl-lH-indazol-5-yl)amino]-2-(prop- 2-en-l-yl)-lH,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (138 mg). After purification by reversed-phase chromatography the pure fractions were loaded on an SCX-2 cartridge (1 g, 0.67 mEq/g), washed with MeOH, eluted with NFE/MeOH (1.4 M) and concentrated under reduced pressure. Yield: 46 mg free base (32%) as a brown solid. LCMS m/z Found: 549.0, MH .

[0274] ¹HNMR (400 MHz, dmso) 5 10.34 (s, 1H), 8.89 (s, 1H), 8.28 (s, 2H), 7.98 (s, 1H), 7.74 (d, J = 3.6 Hz, 1H), 7.61 (d, J = 8.7 Hz, 2H), 7.57 (d, J = 8.9 Hz, 1H), 6.59 (d, J = 3.5 Hz, 1H), 5.73 (ddt, J = 16.2, 10.6, 5.7 Hz, 1H), 5.00 (d, J = 10.3 Hz, 1H), 4.96 - 4.89 (m, 1H), 4.86 (d, J = 17.3 Hz, 1H), 4.64 (s, 2H). 4.02 (s. 3H), 2.73 - 2.54 (m. 4H), 2.30 (s, 3H), 2.23 - 2.04 (m, 2H), 2.03 - 1.89 (m, 2H), 1.89 - 1.77 (m, 1H), 1.75 - 1.56 (m, 1H).

[0275] Example 27. Synthesis of6-[(l-methyl-lH-indazol-5-yl)amino]-l-{l-[(3S)-l- methylpyrrolidin-3-yl]-lH-pyrazolo[3,4-b]pyridin-6-yl}-2-(prop-2-en-l-yl)-lH,2H,3H- pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid (Compound 126)

[0276] Prepared using a method similar to that employed in Example 15 using 6-[(l-methyl- lH-indazol-5-yl)amino]-2-(prop-2-en-l-yl)-l-{l-[(3S)-pyrrolidin-3-yl]-lH-pyrazolo[3,4- b]pyridin-6-yl}-lH,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (87 mg, crude). Yield: 51 mg TFA salt (47%) as a pink powder. LCMS m/z Found: 522.0, MH⁺.

[0277] ¹H NMR (400 MHz, dmso) 6 10.52 (s. 1H), 10.13 (s, 1H). 8.94 (s, 1H). 8.58 (d, J = 7.7 Hz, 1H), 8.36 (s, 1H), 8.30 (s, 1H), 8.08 (s, 1H), 7.95 (d, J= 8.3 Hz, 1H), 7.62 (s, 2H), 5.92 - 5.65 (m, 2H), 5.02 (d, J= 10.4 Hz, 1H), 4.90 (d, J= 16.9 Hz, 1H), 4.77 (s, 2H), 4.22 - 4.11 (m, 1H), 4.05 (s, 3H), 4.01 - 3.74 (m, 2H), 3.71 - 3.26 (m. 2H), 2.98 (d, J = 38.5 Hz, 3H), 2.81 - 2.70 (m, 1H), 2.45 - 2.29 (m, 1H). Mixture of two conformers 1 : 1.

[0278] Example 28. Synthesis of 6-[(l-methyl-lH-indazol-5-yl)amino]-l-{l-[(3R)-l- methylpyrrolidin-3-yl]-lH-pyrazolo[3,4-b]pyridin-6-yl]-2-(prop-2-en-l-yl)-lH,2H,3H- pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid (Compound 127)

[0279] Prepared using a method similar to that employed in Example 15 using 6-[(l-methyl- lH-indazol-5-yl)amino]-2-(prop-2-en-l-yl)-l-{l-[(3R)-pyrrolidin-3-yl]-lH-pyrazolo[3,4- b]pyridin-6-yl}-lH,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (85 mg, crude). Yield: 44 mg TFA salt (41%) as a pink powder. LCMS m/z Found: 522.0, MH⁺.

[0280] ¹H NMR (400 MHz, dmso) 5 10.51 (s, 1H), 10.13 (s, 1H). 8.94 (s, 1H), 8.58 (d, J = 7.3 Hz. 1H), 8.35 (s, 1H), 8.31 (s, 1H), 8.08 (s, 1H). 7.95 (d, J= 7.9 Hz, 1H). 7.62 (s, 2H). 5.92 - 5.63 (m, 2H), 5.02 (d, J= 9.9 Hz, 1H), 4.90 (d, J= 17.3 Hz, 1H), 4.76 (s, 2H), 4.22 - 4.10 (m, 1H), 4.05 (s, 3H), 4.00 - 3.72 (m, 2H), 3.71 - 3.26 (m. 2H), 2.99 (d, J = 38.6 Hz, 3H), 2.81 - 2.68 (m, 1H), 2.46 - 2.21 (m, 1H). Mixture of two conformers 1 : 1.

[0281] Example 29. WeelA Kinase Binding Assay

[0282] We used a LanthaScreen Europium (Eu) Kinase Binding Assay to determine inhibitor affinities (IC5O) to WeelA kinase. The assay utilizes an Alexa Fluor 647-labeled ATP competitive kinase tracer that binds to the ATP binding site of a GST-tagged WeelA kinase, while a europium (Eu) labeled antibody binds to the GST tag. The proximity of fluorescently labeled kinase tracer and europium (Eu) donor fluorophore antibody leads to fluorescence resonance energy transfer (FRET) to the fluorescence label (acceptor) upon excitation of the Eu (donor). Displacement of tracer from the ATP-binding site by a test compound disturbs the proximity between both labels thus lowering the FRET.

[0283] This time resolved-FRET binding assay was performed in white 384-well low volume plates (Greiner, cat # 784075), at room temperature in kinase buffer A (KBA; Invitrogen cat# PV3189), consisting of 50 rnM HEPES-NaOH (pH 7.5), 0.01 % Brij-35, 10 rnM MgCl₂, and 1 mM EGTA. 5 pL of compound (diluted in reaction buffer, to 1 % DMSO) were added to vanous wells in the plate, followed by 5 pL each of recombinant human WeelA kinase (full length Weel kinase was expressed by baculovirus in insect cells using aN-terminal GST tag (MW: 99.1 kDa) (Invitrogen cat# PV3817) and LanthaScreen Eu-anti-GST antibody (Invitrogen, cat# PV5594). After this, 5 pL of kinase tracer 178 (Invitrogen. cat# PV5593) was added to the plate and the plate was incubated for 60 minutes at room temperature. The final assay conditions in each well were: 30 nM tracer 178, 5 nM Weel A kinase and 2 nM Eu- labeled antibody in total assay volume of 15 pL. An Envision 2104 (Perkin-Elmer) Plate Reader with the following time-resolve fluorescence setting was used for performing LanthaScreen kinase binding assay.

• Excitation 320 nm (30 nm bandpass)

• Kinase Tracer Emission 665 nm (10 nm bandpass)

• LanthaScreen Eu-anti-Tag Antibody Emission 615 nm (10 nm bandpass)

• Dichroic Mirror Instrument dependent

• Delay Time 100 ps

• Integration Time 200 ps

[0284] To calculate the emission ratio, the acceptor/tracer emission (665 nM) was divided by the antibody/ donor emission (615 nM) and the average of duplicate measurement was used for calculations. Data plotting, analysis of binding, and curve fitting was done using Excel add- in XLfit version 5.5.0.5 (IDBS, Guildford, United Kingdom). Results are presented in Table 2, below, where compounds having an IC50 less than or equal to 10 nM are represented as “A”; compounds having an TC₅₀ greater than 10 nM but less than or equal to 100 nM are represented as "B”; compounds having an IC50 greater than 100 nM but less than or equal to 500 11M are represented as “C”; and compounds having an IC50 greater than 500 nM are represented as “D”. Table 2. Weel Binding IC50 Values for Exemplary Compounds

[0285] Example 30. Mytl kinase Binding Assay

[0286] We used a LanthaScreen Europium (Eu) Kinase Binding Assay to determine the binding affinities (IC50) of compounds to Mytl kinase. The assay was identical to the Weel A kinase binding assay disclosed in Example 4, except for the use of recombinant PMYT-1 (full length PMYT-1 was expressed by baculovirus in insect cells using a N- terminal GST tag (MW: 80.8 kDa) (Invitrogen cat# A30984) and LanthaScreen Eu-anti-GST antibody (Invitrogen, cat# PV5594) in place of recombinant human Weel A kinase and the final concentration of reagents in the assay. The final assay conditions were: 3 nM tracer 178, 2.5 nM PMYT-1 and 1 nM Eu-labeled antibody in total assay volume of 15 pL. Results are presented in Table 2, below, where compounds having an IC50 less than or equal to 20 nM are represented as ”A"; compounds having an IC50 greater than 20 nM but less than or equal to 100 nM are represented as “B”; compounds having an IC50 greater than 100 nM but less than or equal to 500 nM are represented as “C”; and compounds having an IC50 greater than 500 nM but less than 5 pM are represented as “D”.

Table 3. Mytl Kinase Binding IC50 Values for Exemplary Compounds

[0287] Example 31. WeelA kinase and Mytl kinase Cellular Assays

[0288] Cell culture

[0289] ACHN renal cell carcinoma cell line and A427 lung cancer cell line (ATCC) were cultured in Minimum Essential Medium Eagle supplemented with 10% fetal calf serum (Sigma), 1% Penicillin-Streptomycin and lOmM EIEPES buffer (Ely Clone). NIELOVCAR-3 (OVCAR3) ovarian cancer cell line (ATCC) was cultured in RPMI-1640 medium supplemented with 0.01 mg/ml insulin, 20% fetal calf serum 1% Penicillin-Streptomycin and lOmM HEPES buffer. Cell cultures were kept in a humidified incubator at 37° C and 5% CO2. Cells were routinely tested for Mycoplasma contamination.

[0290] AlphaLISA assay

[0291] For target engagement assessment of Mytl kinase inhibition, quantification of Cdkl phosphorylated on threonine 14 was detected using the

AlphaLISA® Sur eFire® Ultra™ Human Phospho-CDKl (Thrl4) assay (Perkin Elmer). ACHN cells were seeded into tissue culture treated 96-well plates (VWR) to a density of 20,000 cells per well. 24h post seeding cells were treated for 4h with compounds at concentrations ranging from 7 to 5000 nM. For Compound 126, target engagement of Mytl kinase was also quantified in OVCAR3 ovarian carcinoma cells and in A427 lung carcinoma cells seeded to a density of 14,000 and 15,000 cells per well respectively. OVCAR3 and A427 cells were treated for 4h with compounds at concentrations ranging from 0.017 to 1000 nM. Cells were washed with PBS and lysed in 50 pl AlphaLISA lysis buffer before freezing at -80 C. Ten pl of the lysate was transferred to 384 well plates and incubated with AlphaLISA donor and acceptor beads according to the manufacturer’s instructions. Dose response curves and EC₅₀ values were calculated and visualized using GraphPad Prism version 9.

[0292] Results are presented in Table 4 below where compounds having tin EC₅₀ less than or equal to 200 nM are represented as “A”; compounds having an EC₅₀ greater than 200 nM but less than or equal to 500 nM are represented as “B”; compounds having an EC₅₀ greater than 500 nM but less than or equal to 1,000 nM are represented as “C”; and compounds hav ing an EC₅₀ greater than 1 ,000 nM are represented as “D”. Results are presented for ACHN cells unless otherwise indicated.

Table 4. CDK1 Thrl4 Phosphorylation EC₅₀ Values for Exemplary Compounds

¹ Result in OVCAR3 cells.

² Results in A427 cells.

[0293] High-content Imaging of pCdkl/2 Y¹⁵

[0294] For target engagement assessment of Weel A kinase inhibition, high-content imaging of ACHN renal cell carcinoma cells was used for quantification of CDK1/2 phosphorylated on tyrosine 15 by immunofluorescence (“IF”). 20,000 cells per well were seeded in tissue culture treated 96 well plates and treated with compounds at concentrations ranging from 7 to 5,000 nM or from 0.3 to 200 nM for 4h. For Compound 126, target engagement of Weel A kinase inhibition was also quantified in OVCAR3 ovarian carcinoma cells and in A427 lung carcinoma cells seeded to a density of 14,000 and 15,000 cells per well respectively.

OVCAR3 and A427 cells were treated for 4h with compounds at concentrations ranging from 0.017 to 1000 nM. Cells were fixated for 15 minutes in 4% paraformaldehyde solution, washed in PBS and permeabilized using 0.2% Triton X-100. Blocking was performed for Ih at RT using Blocker FL Fluorescent Blocking Buffer (Thermo Scientific), thereafter cells were incubated with primary antibody (rabbit anti-pCDKl Y¹⁵, 44539, Cell Signaling Technology) at 4°C ON. Alexa Fluor Plus 647 labelled goat anti-rabbit (A32733, Thermo Scientific) was used as secondary antibody. After washing in PBS, nuclei were stained with DAPI solution for 10 min at RT. protected from light. Cells were imaged using an ImageXpress Pico automated imaging system and CellReporter Xpress software (Molecular Devices). The percentage of cells with nuclear positivity for pCdkl Y¹⁵ compared to DMSO control was determined for each well and drug dose response curves and EC50 values were calculated and visualized using GraphPad Prism.

[0295] Results are presented in Table 5, below, where compounds having an EC50 less than or equal to 100 nM are represented as “A”; compounds having an EC 50 greater than 100 nM but less than or equal to 500 nM are represented as “B”; compounds having an EC₅₀ greater than 500 nM but less than or equal to 1000 nM are represented as “C’; and compounds having an EC50 greater than 1000 nM are represented as 'D' denotes that the compound was only tested at a range of 0.3 to 200 nM and had a calculated EC50 of greater than 200 nM. Results are presented for ACEIN cells unless otherwise indicated.

Table 5. CDK1 Tyrl5 Phosphorylation EC₅₀ Values for Exemplary Compounds

¹ Result in OVCAR3 cells.

² Results in A427 cells.

Cell Titer Glo viability assay

[0296] ACHN cells were seeded at a density of 400 cells per well respectively, in tissue culture treated 384 well plates (Coming Costar). After overnight incubation, cells were treated with compounds at concentrations ranging from 17 to 10 000 nM. Viability compared to untreated control was assessed at 144 h using Cell Titer Gio assay 2.0 (Promega). Results are presented in Table 6, below, where compounds having an absolute EC50 less than or equal to 100 nM are represented as “A”; compounds having an EC₅₀ greater than 100 nM but less than or equal io 500 nM are represented as “B”, compounds having an EC₅₀ greater than 500 nM but less than or equal to 1000 nM are represented as “C”; and compounds having an EC₅₀ greater than 1000 nM are represented as “D”.

Table 6. Viability Data in ACHN Cells for Exemplary Compounds

[0297] The relevant teachings of all patents, published applications and references cited herein are incorporated by reference in their entirety.

[0298] Furthermore, the invention encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms from one or more of the listed claims are introduced into another claim. For example, any claim that is dependent on another claim can be modified to include one or more limitations found in any other claim that is dependent on the same base claim. Where elements are presented as lists, e.g, in Markush group format, each subgroup of the elements is also disclosed, and any element(s) can be removed from the group. It should it be understood that, in general, where the invention, or aspects of the invention, is/are referred to as comprising particular elements and/or features, certain embodiments of the invention or aspects of the invention consist, or consist essentially of, such elements and/or features. For purposes of simplicity , those embodiments have not been specifically set forth in haec verba herein. It is also noted that the terms “comprising” and “containing” are intended to be open and permit the inclusion of additional elements or steps. Where ranges are given, endpoints are included. Furthermore, unless otherw ise indicated or otherwise evident from the context and understanding of one of ordinary skill in the art, values that are expressed as ranges can assume any specific value or sub-range within the stated ranges in different embodiments of the invention, to the tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise. [0299] Without further description, it is believed that one of ordinary skill in the art can, using the preceding description and the illustrative examples, make and utilize the compounds of the present invention and practice the claimed methods. It should be understood that the foregoing discussion and examples merely present a detailed description of certain preferred embodiments. It will be apparent to those of ordinary skill in the art that various modifications and equivalents can be made without departing from the spirit and scope of the invention.

Claims

1. A compound having structural formula I: solvate, enantiomer, tautomer, or diastereomer thereof, or a pharmaceutically acceptable salt of any of the foregoing wherein:

X is N or C(R);

Y is N or C(R);

Z is N or CH;

R¹ is a 4-8 membered saturated heterocyclyl comprising one ring nitrogen atom and optionally comprising a second ring heteroatom selected from N, O, or S. wherein R¹ is optionally substituted on the one ring nitrogen atom with C₁-C₄ alkyl optionally further substituted with one or more substituents independently selected from halo, -OH, -SO₂-C₁-C₄ alkyl, and -O-C₁-C₄ alkyd; and

R² is C₁-C₄ alkyl, C2-C4 alkenyl, or C3-C4 cycloalky 1.

2. The compound of claim 1, wherein Z is CH.

3. The compound of claim 1 or 2, wherein R² is -CH2-CH=CH2.

4. The compound of any one of claims 1-3, wherein R¹ is wherein R³ is hydrogen or C₁-C₄ alkyd optionally substituted with one or more substituents independently selected from halo. -OH, -SO₂-C₁-C₄ alkyl, and -O-C₁-C₄ alkyd.

5. The compound of claim 4 having structural formula II: solvate, enantiomer, tautomer, or diastereomer thereof, or a pharmaceutically acceptable salt of any of the foregoing wherein

X is N or CH;

Y is N or CH;

X and Y are not simultaneously N; and R³ is hydrogen or methyl.

6. The compound of any one of claims 1-5. wherein X is N and Y is CH.

7. The compound of any one of claims 1-5, wherein X is CH and Y is N.

8. The compound of any one of claims 1-5. wherein X is CH and Y is CH.

9. The compound of any one of claims 1-8, wherein R¹ is:

10. The compound of claim 1, wherein the compound is any one of the following compounds:

11. A pharmaceutical composition comprising an effective amount of a compound of any one of claims 1-10; and a pharmaceutically acceptable carrier.

12. A method of inhibiting both Weel A kinase and Mytl kinase activity in a subject comprising the step of administering to the subject an effective amount of a compound of any one of claims 1-10, or a composition of claim 11.

13. A method of treating a subject suffering from a cancer or other disordered cell growth characterized by both aberrant Weel A kinase activity and aberrant Mytl kinase activity comprising the step of administering to the subject an effective amount of a compound of any one of claims 1-6, or a composition of claim 7.

14. The method of claim 13, wherein the cancer is a brain cancer, a cervicocerebral cancer, a cardiac cancer, a gastrointestinal cancer, an esophageal cancer, a thyroid cancer, a small cell cancer, a non-small cell cancer, a breast cancer, a lung cancer, a stomach cancer, a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms' cancer, a skin cancer, malignant melanoma, a neuroblastoma, an osteosarcoma, an Ewing's tumor, a soft part sarcoma, an acute leukemia, a chronic lymphatic leukemia, a chronic myelocytic leukemia, polycythemia vera, a malignant lymphoma, multiple myeloma, a Hodgkin's lymphoma, or a non-Hodgkin’s lymphoma.

15. The method of claim 14, wherein the subject is suffering from a uterine serous carcinoma or a renal cancer.