Pyrazolopyrimidinone compounds for use in methods of inhibiting WEE1 A kinase

Abstract

A pyrazolopyrimidinone compound of formula I: (I), a method for inhibiting a Wee1A kinase or both a Wee1A kinase and a Myt1 kinase.

Description

Background Technology

[0001] Cells are constantly challenged by endogenous and exogenous factors that can affect DNA integrity. To maintain genome stability and prevent the unwanted proliferation of damaged DNA, cells have established an organized signaling network that recognizes DNA damage and halts the cell cycle to properly repair DNA before resuming DNA replication or cell division. The DNA damage response and the cell cycle are closely linked through several cell cycle checkpoints, which are crucial control steps for maintaining genome integrity.

[0002] Cancer cells often possess defective G1 / S checkpoints, typically due to impaired p53 activity caused by mutations or deletions, or by inactivation of viral oncoproteins. Therefore, cancer cells heavily rely on other cell cycle checkpoints, including the G2 / M checkpoint, to avoid the accumulation of harmful DNA damage and mitotic catastrophe. Consequently, it is hypothesized that cancer cells are particularly vulnerable to inhibition by proteins that protect cells from mitosis. (Matheson, CJ et al.) Trends Pharmacol Sci 37, 872-881 (2016).

[0003] Wee1A kinase is a tyrosine kinase belonging to the Wee1 kinase family, which includes Wee1A kinase, Wee1B kinase, and Myt1 kinase. (Rorà, AGL et al.) J Hematol Oncol 13, 126 (2020). The main role of this kinase family is to regulate cell cycle progression and entry into mitosis (Wee1A kinase and Myt1 kinase) or meiosis (Wee1B kinase). The key complex regulating mitotic entry is the Cdk1 / cyclin B1 complex, also known as the mitosis-promoting factor. Wee1A kinase regulates mitotic entry by activating the inhibitory tyrosine 15 site (Y... 15 Phosphorylation of Cdk1 on the β-cell kinase limits the activity of the Cdk1 / cyclin B1 complex. Therefore, inhibiting Wee1A kinase can prevent the inhibitory Y-cell kinase from reaching the β-cell kinase. 15 Phosphorylation effectively promotes the activity of the Cdk1 / cyclin B1 complex. Premature activation of the Cdk1 / cyclin B1 complex can lead to premature mitosis in the presence of unrepaired DNA damage, ultimately resulting in mitotic catastrophe and cell death.

[0004] In addition to its established role in regulating mitotic entry at the G2 / M checkpoint, Wee1A kinase has also been proposed to play an important role at the S intracellular checkpoint by limiting Cdk2 activity. (Elbæk, CR et al.) Cell Reports38, 110261 (2022); Elbæk, CR et al. Mutat Res Fundam Mol Mech Mutagen 819-820, 111694 (2020). Cdk2 activity is regulated by Wee1A kinase in the same manner as Cdk1, via phosphorylation at tyrosine 15. Cdk2 is the major Cdk driving DNA replication, and inhibition of Wee1A kinase leads to excessive DNA replication, resulting in depletion of the nucleotide pool and degradation of the ribonucleotide reductase subunit RRM2 (Pfister, SX et al.). Cancer Cell 28, 557-568 (2015). Pfister et al. demonstrated that Wee1A kinase inhibition selectively kills H3K through dNTP starvation induced by RRM2 depletion. 36 ME3-deficient cancer cells. Histone methyltransferase SETD2 catalyzes H3K. 36 me3, thereby promoting RRM2 expression and dNTP synthesis. Inactivation of the SETD2 gene is common in clear cell renal cell carcinoma (ccRCC), and therefore may be sensitive to Wee1A kinase inhibition. A phase II trial is currently testing AZD1775 (NCT03284385) in SETD2-deficient solid tumors.

[0005] It has also been proposed that Wee1A kinase plays a role in controlling histone stoichiometry in late S phase by phosphorylating the core histone H2B at tyrosine 37. (Koh, S.-B) Cell Signal 94, 110310 (2022).

[0006] Cancers associated with high-risk human papillomavirus (HPV), such as head and neck squamous cell carcinoma (HNSCC), show increased sensitivity to Wee1A kinase inhibitors. (Diab, A. et al.) Proc National Acad Sci 117, 28287-28296 (2020).

[0007] Several Wee1A kinase inhibitors are currently being tested in clinical trials (Bukhari, AB et al.). Frontiers Oncol 12, 828684 (2022)) and has shown activity in many indications. The phase II study of the Wee1A kinase inhibitor AZD1775 (adavosertib) showed promising results in women with serous uterine carcinoma. Liu, JF et al. J Clin Oncol 39, 1531-1539 (2021). With RAS / TP 53Adatimb also showed efficacy compared to active surveillance of metastatic colorectal cancer with mutations. (Seligmann, JF et al.) J Clin Oncol 39, 3705-3715 (2021). In a phase 1b trial of 18 patients with platinum-resistant ovarian cancer, the combination of ZN-c3 (azenosertib) and paclitaxel resulted in an objective response rate (ORR) of 50% (…). J Clin Oncol 41, 2023 (Supplement 16; Abstract) 5513))。 Given the encouraging signs of Wee1A kinase inhibition in terms of clinical activity, there is an urgent need for novel Wee1A kinase inhibitors with improved potency and selectivity, as well as compounds that simultaneously inhibit Wee1A kinase and Myt1 kinase, to maximize the efficacy potential of this class of targets. Summary of the Invention

[0008] In some embodiments, this disclosure provides compounds of formula I:

[0009] (I),

[0010] Or a pharmaceutically acceptable salt thereof, wherein X, Y, Z, R 1 R 2 R 3 R 4 R 8 , n and m are each defined as follows and as described herein.

[0011] In some embodiments, this disclosure provides a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0012] In some embodiments, this disclosure provides a method for inhibiting Wee1A kinase in a patient or biological sample, the method comprising administering a compound of formula I or a pharmaceutically acceptable salt thereof to a patient, or contacting a biological sample with a compound of formula I or a pharmaceutically acceptable salt thereof.

[0013] In some embodiments, this disclosure provides a method for inhibiting both Wee1A kinase and Myt1 kinase in a patient or biological sample, the method comprising administering a compound of formula I or a pharmaceutically acceptable salt thereof to a patient, or contacting a biological sample with a compound of formula I or a pharmaceutically acceptable salt thereof.

[0014] In some embodiments, this disclosure provides a method for treating a disease or condition associated with Wee1A kinase, the method comprising administering a compound of formula I or a pharmaceutically acceptable salt thereof to a patient in need.

[0015] In some embodiments, this disclosure provides a method for treating a disease or condition associated with both Wee1A kinase and Myt1 kinase, the method comprising administering to a patient in need a compound of formula I having dual activity or a pharmaceutically acceptable salt thereof.

[0016] In some implementations of this kind, the disease or condition associated with Wee1A kinase is cancer. In some implementations of this kind, the disease or condition associated with both Wee1A kinase and Myt1 kinase is cancer. In some implementations, the cancer is selected from brain cancer, cervical brain cancer, heart cancer, gastrointestinal cancer, esophageal cancer, thyroid cancer, small cell carcinoma, non-small cell carcinoma, breast cancer, lung cancer, stomach cancer, gallbladder / choleduct cancer, liver cancer, pancreatic cancer, colon cancer, rectal cancer, ovarian cancer, choriocarcinoma, endometrial cancer, cervical cancer, renal pelvis / ureter cancer, bladder cancer, prostate cancer, penile cancer, testicular cancer, fetal cancer, Wilms' cancer, skin cancer, malignant melanoma, neuroblastoma, osteosarcoma, Ewing's tumor, soft tissue sarcoma, acute leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, polycythemia vera, malignant lymphoma, multiple myeloma, Hodgkin's lymphoma, and non-Hodgkin's lymphoma. Detailed Implementation

[0017] 1. Overview of compounds disclosed herein

[0018] In some embodiments, this disclosure provides inhibitors of Wee1A kinase. In some embodiments, such compounds include those of the formulas described herein or pharmaceutically acceptable salts thereof, wherein each variable is as defined and described herein.

[0019] In some embodiments, this disclosure provides compounds having the structural formula AA:

[0020] (AA),

[0021] Or its solvates, enantiomers, tautomers or diastereomers, or pharmaceutically acceptable salts of any of the foregoing substances, wherein:

[0022] X, Y, and Z are each independently CH or N;

[0023] R 1 It is -O-, -NH-, or -N(C1-C3 alkyl)-;

[0024] Each R 2 Independently, it is a fluorinated group, -CN, unsubstituted C1-C5 alkyl, fluorinated C1-C5 alkyl, or cyano-substituted C1-C5 alkyl, wherein the two R groups bonded to the same carbon atom are...2 Optionally, they can be combined to form a spirocyclic fused C3-C7 cycloalkyl ring, or two R atoms bonded to different carbon atoms. 2 Optionally, they can be combined to form a bridged or fused C3-C5 cycloalkyl ring, or an R 2 and R 8 They can be optionally combined to form bridging or fused 3- to 5-membered rings;

[0025] R 3 It is a -C1-C3 alkyl group or -CH2-CH=CH2;

[0026] R 4 yes , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or ,in" " indicates R 4Connection points with compounds;

[0027] Each R 5 and each R 6 Independently, it is hydrogen, a halogroup, -CN, a -C1-C4 alkyl group optionally substituted with a halogroup, an -O-(C1-C4 alkyl group optionally substituted with a halogroup, an -O-(C1-C4 alkyl group substituted with a C3-C6 cycloalkyl group, an N-linked saturated 3-7 membered heterocyclic group, a 5-6 membered heteroaryl group, or a phenyl group, wherein there are no more than two R groups. 6 It is not hydrogen; the R on the adjacent ring atoms 5 and R 6 Optionally together form with R 4 Fused 4-7 membered saturated heterocycles or cycloalkyl rings;

[0028] R 7 It is hydrogen, -C1-C4 alkyl, -C1-C4 alkylene, -O-C1-C4 alkyl, -C(O)-C1-C4 alkyl, or C3-C6 cycloalkyl, wherein R 7 Any C1-C4 alkyl or C1-C4 alkylene moiety may optionally be substituted by one or more substituents independently selected from halogen and -CN;

[0029] R 8 It is hydrogen, -C1-C4 alkyl, or 4-6 membered saturated heterocycle;

[0030] R 9 It is hydrogen, a halogroup, -CN, a -C1-C4 alkyl group optionally substituted with a halogroup, an -O-C1-C4 alkyl group optionally substituted with a halogroup, or a phenyl group optionally substituted;

[0031] m is 0, 1, 2, 3 or 4;

[0032] n is 0, 1, 2, or 3; and

[0033] m+n is 1, 2, 3, or 4.

[0034] Any hydrogen atom may be optionally replaced by deuterium.

[0035] In some embodiments, this disclosure provides compounds having structural formula A:

[0036] (A),

[0037] Or its solvates, enantiomers, tautomers or diastereomers, or pharmaceutically acceptable salts of any of the foregoing substances, wherein:

[0038] X, Y, and Z are each independently CH or N;

[0039] R 1It is -O-, -NH-, or -N(C1-C3 alkyl)-;

[0040] Each R 2 Independently, it is a fluorinated group, -CN, unsubstituted C1-C5 alkyl, fluorinated C1-C5 alkyl, or cyano-substituted C1-C5 alkyl, wherein the two R groups bonded to the same carbon atom are... 2 Optionally, they can be combined to form a spirocyclic fused C3-C7 cycloalkyl ring, or two R atoms bonded to different carbon atoms. 2 Optionally, they can be combined to form a bridged C3-C5 cycloalkyl ring, or an R 2 and R 8 They can be arbitrarily combined to form bridging 3-5 element rings;

[0041] R 3 It is a -C1-C3 alkyl group or -CH2-CH=CH2;

[0042] R 4 yes , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or ,in" " indicates R 4 Connection points with compounds;

[0043] Each R 5 and each R 6 Independently, it is hydrogen, a halogroup, -CN, a -C1-C4 alkyl group optionally substituted with a halogroup, an -O-(C1-C4 alkyl group optionally substituted with a halogroup, an -O-(C1-C4 alkyl group substituted with a C3-C6 cycloalkyl group, an N-linked saturated 3-7 membered heterocyclic group, a 5-6 membered heteroaryl group, or a phenyl group, wherein there are no more than two R groups. 6 It is not hydrogen; the R on the adjacent ring atoms 5 and R 6 Optionally together form with R 4 Fused 4-7 membered saturated heterocycles or cycloalkyl rings;

[0044] R 7 It is hydrogen, -C1-C4 alkyl, -C1-C4 alkylene, -O-C1-C4 alkyl, -C(O)-C1-C4 alkyl, or C3-C6 cycloalkyl, wherein R 7 Any C1-C4 alkyl or C1-C4 alkylene moiety may optionally be substituted by one or more substituents independently selected from halogen and -CN;

[0045] R 8 It is hydrogen, -C1-C4 alkyl, or 4-6 membered saturated heterocycle;

[0046] R 9 It is hydrogen, a halogroup, -CN, a -C1-C4 alkyl group optionally substituted with a halogroup, an -O-C1-C4 alkyl group optionally substituted with a halogroup, or a phenyl group optionally substituted;

[0047] m is 0, 1, 2, 3 or 4;

[0048] n is 0, 1, 2, or 3; and

[0049] m+n is 1, 2, 3, or 4.

[0050] Any hydrogen atom may be optionally replaced by deuterium.

[0051] In some embodiments, this disclosure provides compounds having structural formula I:

[0052] (I),

[0053] Or its solvates, enantiomers, tautomers or diastereomers, or pharmaceutically acceptable salts of any of the foregoing substances, wherein:

[0054] X, Y, and Z are each independently CH or N;

[0055] R 1 It is -O-, -NH-, or -N(C1-C3 alkyl)-;

[0056] Each R 2 Independently, it is a fluorinated group, -CN, unsubstituted C1-C5 alkyl, fluorinated C1-C5 alkyl, or cyano-substituted C1-C5 alkyl, wherein the two R groups bonded to the same carbon atom are... 2 Optionally, they can be combined to form a spirocyclic fused C3-C7 cycloalkyl ring, or two R atoms bonded to different carbon atoms. 2 Optionally, they can be combined to form a bridged C3-C5 cycloalkyl ring, or an R 2 and R 8 They can be arbitrarily combined to form bridging 3-5 element rings;

[0057] R 3 It is a -C1-C3 alkyl group or -CH2-CH=CH2;

[0058] R 4 yes , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or ,in" " indicates R 4 Connection points with compounds;

[0059] Each R 5 and each R 6 Independently, it is hydrogen, a halogroup, -CN, a -C1-C4 alkyl group optionally substituted with a halogroup, or an -O-(C1-C4 alkyl group optionally substituted with a halogroup), wherein there are no more than two R groups. 6 It is not hydrogen; the R on the adjacent ring atoms 5 and R 6 Optionally together form with R 4 Fused 4-7 membered saturated heterocycles or cycloalkyl rings;

[0060] R 7 It is hydrogen, -C1-C4 alkyl or -C1-C4 alkylene-O-C1-C4 alkyl, wherein R 7 Any C1-C4 alkyl or C1-C4 alkylene moiety may optionally be substituted by one or more substituents independently selected from halogen and -CN;

[0061] R 8 It is hydrogen or -C1-C4 alkyl;

[0062] R 9 It is hydrogen, a halogroup, -CN, a -C1-C4 alkyl group optionally substituted with a halogroup, an -O-C1-C4 alkyl group optionally substituted with a halogroup, or a phenyl group optionally substituted;

[0063] m is 0 or 1;

[0064] n is 0, 1, 2, or 3; and

[0065] m and n are not both 0.

[0066] 2. Compounds and definitions

[0067] The compounds disclosed herein include those generally described above, and are further illustrated by the categories, subclasses, and species disclosed herein. Unless otherwise stated, the following definitions shall apply as used herein. For the purposes of this disclosure, chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th edition. Furthermore, the general principles of organic chemistry are described in “Organic Chemistry”, Thomas Sorrell, University Science Books, Sausalito: 1999 and “March's Advanced Organic Chemistry”, 5th edition, eds. Smith, MB and March, J., John Wiley & Sons, New York: 2001, the entire contents of which are hereby incorporated by reference.

[0068] As used herein, the term "aliphatic" or "aliphatic group" means a fully saturated or branched, substituted or unsubstituted hydrocarbon chain containing one or more unsaturated units (i.e., unbranched) or one or more unsaturated units, or a fully saturated or bicyclic hydrocarbon (also referred to herein as "carbocyclic", "aliphatic", or "cycloalkyl") containing one or more unsaturated units but not aromatic, having a single connection point with the rest of the molecule. Unless otherwise stated, an aliphatic group contains 1 to 6 aliphatic carbon atoms. In some embodiments, the aliphatic group contains 1 to 5 aliphatic carbon atoms. In other embodiments, the aliphatic group contains 1 to 4 aliphatic carbon atoms. In still other embodiments, the aliphatic group contains 1 to 3 aliphatic carbon atoms, and in yet another embodiment, the aliphatic group contains 1 to 2 aliphatic carbon atoms. In some embodiments, "aliphatic" (or "carbocyclic" or "cycloalkyl") means a fully saturated or non-aromatic monocyclic C3-C6 hydrocarbon containing one or more unsaturated units but not aromatic, having a single connection point with the rest of the molecule. Suitable aliphatic groups include, but are not limited to, straight-chain or branched substituted or unsubstituted alkyl, alkenyl and alkynyl groups and mixtures thereof, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl.

[0069] As used herein, the term "alkyl" means a fully saturated branched or branched hydrocarbon. Exemplary alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, etc.

[0070] The term "heteroatom" refers to one or more of oxygen, sulfur, nitrogen, phosphorus, or silicon (including any oxidized form of nitrogen, sulfur, phosphorus, or silicon; any quaternized form of basic nitrogen; or a substituted nitrogen of a heterocycle, such as N (e.g., in 3,4-dihydro-2-) H -pyrrole group), NH (as in pyrroleyl group) or NR + (e.g., in N-substituted pyrroleyl groups).

[0071] As used in this article, the term "unsaturated" means that a part has one or more unsaturated units.

[0072] As used herein, the term "partially unsaturated" refers to a moiety comprising at least one double or triple bond. The term "partially unsaturated" is intended to cover moieties having multiple unsaturated sites, but when used to describe rings, it is not intended to include aryl or heteroaryl moieties as defined herein.

[0073] The term "saturated" refers to a portion that does not have double or triple bonds.

[0074] The term "alkylene" refers to a divalent alkyl group. An "alkylene chain" is a polymethylene group, i.e., (CH2). n - where n is a positive integer, preferably 1 to 6, 1 to 4, 1 to 3, 1 to 2, or 2 to 3. The substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are replaced by substituents. Suitable substituents include those that substituted carbon atoms as described below.

[0075] The term "alkenyl" refers to a divalent alkenyl group. A substituted alkenyl chain is a polymethylene group containing at least one double bond, wherein one or more hydrogen atoms are replaced by substituents. Suitable substituents include those that substituted carbon atoms as described below.

[0076] The terms “halogen” and “halogenated group” are used interchangeably and refer to F, Cl, Br or I.

[0077] The term "aryl," used alone or as part of a larger portion of "aralkyl," "ararylalkoxy," or "aryloxyalkyl," refers to monocyclic and bicyclic ring systems having a total of five to fourteen ring members, wherein at least one ring in the system is aromatic and each ring in the system contains three to seven ring members. The term "aryl" may be used interchangeably with the term "aryl ring." In certain embodiments of this disclosure, "aryl" refers to an aromatic ring system, including but not limited to phenyl, biphenyl, naphthyl, anthracene, etc., which may have one or more substituents. As used herein, the term "aryl" also includes, within the scope of the term, groups in which an aromatic ring is fused to one or more non-aromatic carbocyclic rings.

[0078] The term "cycloalkyl ring" refers to a fully saturated carbon ring.

[0079] When two rings share a single ring carbon atom, the first ring and the second ring are "spiro-fused". An example of a spiro-fused ring system is: .

[0080] When two rings share two directly connected carbon atoms, the first ring and the second ring are "fused". Examples of fused ring systems are: and .

[0081] When two rings share two carbon atoms separated by at least one ring carbon atom, the first ring is "bridged" to the second ring. Examples of bridged ring systems are: , , and .

[0082] Referring to the number of atoms in a bridging ring, for example, one R 2 and R 8 The formation of bridging 3-5 membered rings refers to the combination of two bridgehead ring atoms and any additional ring atoms between these two bridgehead atoms that form a ring that bridges an existing ring. For example, if the structure... The limited ring is And R 2 and R 8 Together As enumerated in the bridging structure, the number of atoms in the bridging ring is 4.

[0083] The terms "heteroaryl" and "heteroaryl-", such as "heteroarylalkyl" or "heteroarylalkoxy", used alone or as part of a larger part, refer to a group having 5 to 10 ring atoms, preferably 5, 6, or 9 ring atoms; having 6, 10, or 14 π electrons shared in the ring array; and having one to five heteroatoms in addition to carbon atoms. The term "heteroatom" refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, as well as any quaternized form of basic nitrogen. Heteroaryl groups include, but are not limited to, thienyl, furanyl, pyrroleyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiazolyl, pyridyl, pyridinyl, pyridazinyl, indoleazinyl, purine, naphthidyl, and pteridinyl. As used herein, the terms "heteroaryl" and "heteroary-" also include groups in which a heteroaryl ring is fused with one or more aryl rings or heteroaryl rings such that the resulting bicyclic or polycyclic system is entirely aromatic. Non-limiting examples include indolyl, isoindolyl, benzothiopheneyl, benzofuranyl, dibenzofuranyl, indazoleyl, benzimidazolyl, benzothiazolyl, quinolinyl, isoquinolinyl, cenolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinazinyl, carbazoleyl, acridineyl, phenazinyl, phenothiazinyl, and phenotoxazinyl. Heteroaryl groups can be monocyclic or bicyclic. The term "heteroaryl" is used interchangeably with the terms "heteroaryl ring," "heteroaryl group," or "heteroaryl family," any of which includes optionally substituted rings. The term "heteroaryl" refers to an alkyl group substituted with a heteroaryl group, wherein the alkyl and heteroaryl portions are optionally substituted independently.

[0084] As used herein, the terms “heterocycle,” “heterocyclic group,” “heterocyclic group,” and “heterocycle” are used interchangeably and refer to a stable 5- to 7-membered monocyclic or 7- to 10-membered bicyclic heterocyclic portion that is saturated or partially unsaturated and has one or more (preferably one to four) heteroatoms as defined above, in addition to a carbon atom. When referring to the ring atom of a heterocycle, the term “nitrogen” includes substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0 to 3 heteroatoms selected from oxygen, sulfur, or nitrogen, said nitrogen may be N (e.g., in 3,4-dihydro-2H-pyrrole), NH (e.g., in pyrrolealkyl), or +NR (e.g., in N-substituted pyrrolealkyl).

[0085] Heterocycles can be attached to their side groups at any heteroatom or carbon atom that produces a stable structure, and any one of the ring atoms can optionally be substituted. Examples of such saturated or partially unsaturated heterocyclic groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenylpyrrolidinyl, piperidinyl, pyrrolidinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazine, dioxalyl, dioxopentyl, diazazyl, oxonitrile, thioazinotrile, morpholinyl, and quininecycloyl. The terms “heterocycle,” “heterocyclic group,” “heterocyclic ring,” “heterocyclic group,” “heterocyclic moiety,” and “heterocyclic radica” are used interchangeably herein and also include groups in which the heterocyclic ring is fused with one or more aryl, heteroaryl, or cycloaliphatic rings, such as dihydroindolyl, 3H-indolyl, benzodihydropyranyl, phenanthridine, or tetrahydroquinolinyl. The heterocyclic group can be monocyclic or bicyclic. The term "heterocyclic alkyl" refers to an alkyl group substituted with a heterocyclic group, wherein the alkyl and heterocyclic moieties are optionally substituted independently.

[0086] As described herein, the compounds of this disclosure may contain an "optionally substituted" portion. Generally, regardless of whether the term "optionally" is preceding it, the term "substituted" means that one or more hydrogens of the specified portion are replaced by suitable substituents. Unless otherwise specified, the "optionally substituted" group may have suitable substituents at each substituted position of the group, and the substituents may be the same or different at each position when more than one position in any given structure may be substituted by more than one substituent selected from the specified group. The combinations of substituents contemplated in this disclosure are preferably those that result in the formation of stable or chemically viable compounds. As used herein, the term "stable" means a compound that remains substantially unchanged when subjected to conditions permissible for compound preparation, testing, and, in some embodiments, recovery, purification, and use for one or more purposes disclosed herein.

[0087] The term "cyano-substituted C" x -C y Alkyl and fluorinated C x -C y Alkyl (where x and y are each integers) refers to the corresponding alkyl group in which hydrogen is replaced by one or more specified substituents.

[0088] The suitable monovalent substituent on the substituted carbon atom of the "optionally substituted" group is independently a halogen; -(CH2) 0-4 R°;-(CH2) 0-4 OR°;-O(CH2) 0-4 R°、-O-(CH2) 0-4 C(O)OR°;-(CH2) 0-4CH(OR°)2;-(CH2) 0- 4SR°;-(CH2) 0-4 Ph, which can be replaced by R°; -(CH2) 0-4 O(CH2) 0-1 Ph, which can be substituted by R°; -CH=CHPh, which can be substituted by R°; -(CH2) 0-4 O(CH2) 0-1 -Pyridyl group, which can be substituted with R°; -NO2; -CN; -N3; ​​(CH2) 0-4 N(R°)2;-(CH2) 0-4 N(R°)C(O)R°;-N(R°)C(S)R°;-(CH2) 0-4 N(R°)C(O)NR°2; N(R°)C(S)NR°2; -(CH2) 0-4 N(R°)C(O)OR°; -N(R°)N(R°)C(O)R°; N(R°)N(R°)C(O)NR°2; N(R°)N(R°)C(O)OR°; -(CH2) 0-4 C(O)R°;-C(S)R°;-(CH2) 0-4 C(O)OR°;-(CH2) 0-4 C(O)SR°;(CH2) 0-4 C(O)OSiR°3;-(CH2) 0-4 OC(O)R°;-OC(O)(CH2) 0-4 SR°, SC(S)SR°; -(CH2) 0-4 SC(O)R°;-(CH2) 0-4 C(O)NR°2; -C(S)NR°2; -C(S)SR°; -SC(S)SR°, (CH2) 0-4 OC(O)NR°2; C(O)N(OR°)R°; -C(O)C(O)R°; -C(O)CH2C(O)R°; -C(NOR°)R°; (CH2) 0-4 SSR°;-(CH2) 0-4 S(O)2R°;-(CH2) 0-4 S(O)₂OR°;-(CH₂) 0-4 OS(O)2R°;-S(O)2NR°2; (CH2) 0- 4S(O)R°; N(R°)S(O)2NR°2; -N(R°)S(O)2R°; -N(OR°)R°; -C(NH)NR°2; -P(O)2R°; P(O)R°2; OP(O)R°2; -OP(O)(OR°)2; SiR°3; -(C 1-4(linear or branched alkylene)ON(R°)2; or -(C 1-4 (straight-chain or branched alkylene)C(O)ON(R°)2, wherein each R° can be substituted as defined below and is independently hydrogen, C 1-6 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph, -CH2- (5-6 membered heteroaryl ring) or 5-6 membered saturated, partially unsaturated or aryl ring having 0-4 independent heteroatoms selected from nitrogen, oxygen or sulfur, or although defined above, two independently occurring R° together with their intervening atoms form a 3-12 membered saturated, partially unsaturated or aryl monocyclic or bicyclic ring having 0-4 independent heteroatoms selected from nitrogen, oxygen or sulfur, which may be substituted as defined below.

[0089] A suitable monovalent substituent on R° (or a ring formed by two independently occurring R° with their intervening atoms) is independently a halogen, -(CH2). 0-2 R ● -(halogenated R) ● -(CH2) 0-2 OH, -(CH2) 0-2 OR ● -(CH2) 0-2 CH(OR ● )2; O(halogenated R) ● -CN, -N3, -(CH2) 0-2 C(O)R ● -(CH2) 0-2 C(O)OH, -(CH2) 0-2 C(O)OR ● -(CH2) 0-2 SR ● -(CH2) 0-2 SH, -(CH2) 0-2 NH2、-(CH2) 0-2 NHR ● -(CH2) 0-2 NR ● 2, -NO2, -SiR ● 3. -OSiR ● 3. C(O)SR ● 、 -(C 1-4 (linear or branched alkylene)C(O)OR ● or -SSR ● , where each R ● It is either unsubstituted or, when previously "halogenated", substituted by only one or more halogens, and independently selected from C. 1-4 Aliphatic, -CH2Ph, -O(CH2)0-1 Ph or a 5-7 member saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents on the saturated carbon atom of R° include =O and =S.

[0090] Suitable divalent substituents on the saturated carbon atom of the "optionally substituted" group include the following: =O, =S, =NNR. * 2、=NNHC(O)R * =NNHC(O)OR * =NNHS(O)2R * =NR * =NOR * -O(C(R) * 2)) 2-3 O- or -S(C(R) * 2)) 2- 3S-, where each independently occurring R * Selected from hydrogen, C can be substituted as defined below. 1-6 Aliphatic or unsubstituted 5-6 member saturated, partially unsaturated, or aryl rings having 0-4 independent heteroatoms selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents bonded to the ortho-substituted carbon of the "optionally substituted" group include: -O(CR * 2) 2-3 O-, where each independently occurring R * Selected from hydrogen, C can be substituted as defined below. 1-6 Aliphatic or unsubstituted 5-6 saturated, partially unsaturated or aryl rings having 0-4 independent heteroatoms selected from nitrogen, oxygen or sulfur.

[0091] In R * Suitable substituents on aliphatic groups include halogens, -R ● 、(halogenated R) ● ), OH, -OR ● -O (halogenated R) ● -CN, -C(O)OH, -C(O)OR ● -NH2, -NHR ● -NR ● 2 or -NO2, where each R ● It is either unsubstituted or, when previously "halogenated", substituted by only one or more halogens, and is independently C. 1-4 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph or a 5-6 member saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0092] Suitable substituents on the substituted nitrogen of the "optionally substituted" group include -R † -NR † 2. -C(O)R † -C(O)OR † -C(O)C(O)R † -C(O)CH2C(O)R † -S(O)2R † S(O)2NR † 2. -C(S)NR † 2. -C(NH)NR † 2 or -N(R) † )S(O)2R † ; where each R † Independently, it is hydrogen, and the C that can be substituted can be defined as follows. 1-6 Aliphatic, unsubstituted -OPh or substituted 5-6 member saturated, partially unsaturated or aryl rings having 0-4 independently selected heteroatoms chosen from nitrogen, oxygen or sulfur, or, although defined above, two independently occurring R... † Together with its intercalary atoms, it forms an unsubstituted 3-12 saturated, partially unsaturated, or aryl monocyclic or bicyclic ring with 0-4 independent heteroatoms selected from nitrogen, oxygen, or sulfur.

[0093] In R † The aliphatic group and suitable substituents on the 5-6 member saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur are independently halogenated, -R ● 、(halogenated R) ● -OH, -OR ● -O (halogenated R) ● -CN, -C(O)OH, -C(O)OR ● -NH2, -NHR ● -NR ● 2 or NO2, where each R ● It is either unsubstituted or, when previously "halogenated", substituted by only one or more halogens, and is independently C. 1-4 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph or a 5-6 member saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0094] As used herein, the term "pharmaceutically acceptable salt" means that, to the extent of reasonable medical judgment, it is suitable for contact with the tissues of humans and lower animals without undue toxicity, irritation, anaphylactic response, etc., and in proportion to a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in detail by SM Berge et al. in J. Pharmaceutical Sciences, 1977, 66, 1-19 (incorporated herein by reference). Pharmaceutically acceptable salts of the compounds disclosed herein include those derived from suitable inorganic acids and organic acids with bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts formed by an amino group with an inorganic acid (such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid) or with an organic acid (such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid), or salts formed by using other methods used in the art (such as ion exchange). Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, hydrogen sulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, disglucuronate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucono-heptahydrate, glyceryl phosphate, gluconate, hemisulfate, heptahydrate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lacturonate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, dihydroxynaphthalate, pectate, persulfate, 3-phenylpropionate, phosphate, neopentanoate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc.

[0095] Salts derived from suitable bases include alkali metal salts, alkaline earth metal salts, ammonium salts, and N salts. + (C 1-4 Alkyl)4 salts. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Further pharmaceutically acceptable salts include, where appropriate, non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates, and aryl sulfonates.

[0096] Unless otherwise stated, the structures described herein are also intended to include all isomers (e.g., enantiomers, diastereomers, and geometric (or conformations)) of the structure; for example, R and S configurations for each asymmetric center, Z and E double bond isomers, rotational isomers (restricted isomers), and Z and E conformational isomers. Therefore, single stereochemical isomers of the compounds of this disclosure, as well as enantiomers, diastereomers, and mixtures of geometric (or conformations), are within the scope of this disclosure. Unless otherwise stated, all tautomers of the compounds of this disclosure are within the scope of this disclosure. Additionally, unless otherwise stated, the structures described herein are also intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the structures of the present invention (including hydrogen replaced by deuterium or tritium, or carbon replaced by...) 13 C or 14 Compounds enriched with carbon substitution (C-rich carbon-substituted) are within the scope of this disclosure. Such compounds can be used, for example, as probes or therapeutic agents in analytical tools, bioassays, or other applications according to this disclosure.

[0097] The combinations of substituents and variables contemplated in this disclosure are only those that result in the formation of stable compounds. As used herein, the term "stable" means a compound that has sufficient stability to allow manufacture and maintains the integrity of the compound for a sufficient period of time for use for the purposes detailed herein (e.g., therapeutic or prophylactic administration to a subject).

[0098] The listing of chemical groups in the definition of any variable herein includes defining the variable as any single group or a combination of the listed groups. The description of embodiments of variables herein includes the embodiment as any single embodiment or in combination with any other embodiment or part thereof.

[0099] As used herein, the term "biological sample" includes, but is not limited to, cell cultures or extracts thereof; biopsy material obtained from mammals or extracts thereof; and hair, skin, blood, saliva, urine, feces, semen, tears, or other bodily fluids or extracts thereof. Inhibiting the activity of protein kinases (e.g., Wee1A kinase or mutants thereof) in biological samples can be used for a variety of purposes known to those skilled in the art. Examples of such purposes include, but are not limited to, blood transfusions, organ transplantation, biological sample storage, and bioassays.

[0100] As used herein, “disease or condition associated with Wee1A kinase” or alternatively “Wee1A kinase-mediated disease or condition” means any disease or other harmful disorder in which Wee1A kinase or its mutants are known or suspected to play a role.

[0101] As used herein, the term "subject" means mammal and includes both human and animal subjects, such as domesticated animals (e.g., horses, dogs, cats, etc.). The terms "subject" and "patient" are used interchangeably. In some embodiments, "patient" or "subject" means animal, preferably a mammal, and most preferably a human.

[0102] The term "pharmaceutically acceptable carrier, adjuvant, or mediator" refers to a non-toxic carrier, adjuvant, or mediator that does not impair the pharmacological activity of the compound formulated with it. Pharmaceutically acceptable carriers, adjuvants, or mediators 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, buffering substances such as phosphates, glycine, sorbic acid, potassium sorbate, mixtures of partial glycerides of saturated vegetable fatty acids, water, salts, or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, silica gel, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polypropylene block polymers, polyethylene glycol, and lanolin. The amount of the compounds of this disclosure that may be combined with carrier materials to produce compositions of a single dosage form will vary depending on the host being treated, the specific route of administration, etc. Preferably, the provided compositions are formulated such that an inhibitor dose of 0.01 to about 100 mg / kg, or about 0.1 mg / kg to about 50 mg / kg, and more preferably about 1 mg / kg to about 25 mg / kg of subject body weight / day, can be administered to a patient receiving these compositions to obtain the desired therapeutic effect. The amount of the compounds disclosed herein in the composition will also depend on the specific compounds in the composition.

[0103] As used herein, the term "treatment" means the partial or complete reduction, suppression, delay of onset, prevention, improvement, and / or relief of a condition or disorder or one or more symptoms thereof as described herein. In some embodiments, treatment may be administered after one or more symptoms have developed. In some embodiments, the term "treatment" includes preventing or halting the progression of a disease or condition. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered to a susceptible individual before symptoms occur (e.g., based on a history of symptoms and / or based on genetic or other susceptibility factors). Treatment may also continue after symptoms have subsided, for example, to prevent or delay their recurrence. Therefore, in some embodiments, the term "treatment" includes preventing the recurrence or relapse of a disease or condition.

[0104] As used herein, the term "inhibitor" is defined as a compound that binds to and / or inhibits a target protein kinase with measurable affinity. In some embodiments, the inhibitor's IC50 value is... 50And / or the binding constant is less than about 50 µM, less than about 1 µM, 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.

[0105] As used herein, the terms “measurable affinity” and “measurable inhibition” refer to a measurable change in the activity of a sample containing a compound or composition thereof of the present disclosure and an equivalent sample containing a Wee1A kinase or Myt1 kinase (excluding the compound or composition thereof).

[0106] As used herein, the terms “dual inhibitor” and “dual Wee1A kinase / Myt1 kinase inhibitor” are used interchangeably and refer to compounds disclosed herein that meet one or more of the following criteria: 1) Myt1 kinase binding activity IC50. 50 ≤ 100 nM (i.e., rated "A" or "B" in Table 3); or 2) Myt1 kinase target binding EC 50 ≤ 500 nM (i.e., rated "A" or "B" in Table 4).

[0107] 3. Description of exemplary compounds

[0108] In some embodiments, this disclosure provides compounds having the structural formula AA:

[0109] (AA),

[0110] Or its solvates, enantiomers, tautomers or diastereomers, or pharmaceutically acceptable salts of any of the foregoing substances, wherein:

[0111] X, Y, and Z are each independently CH or N;

[0112] R 1 It is -O-, -NH-, or -N(C1-C3 alkyl)-;

[0113] Each R 2 Independently, it is a fluorinated group, -CN, unsubstituted C1-C5 alkyl, fluorinated C1-C5 alkyl, or cyano-substituted C1-C5 alkyl, wherein the two R groups bonded to the same carbon atom are... 2 Optionally, they can be combined to form a spirocyclic fused C3-C7 cycloalkyl ring, or two R atoms bonded to different carbon atoms. 2 Optionally, they can be combined to form a bridged or fused C3-C5 cycloalkyl ring, or an R 2 and R 8 They can be optionally combined to form bridging or fused 3- to 5-membered rings;

[0114] R3 It is a -C1-C3 alkyl group or -CH2-CH=CH2;

[0115] R 4 yes , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or ,in" " indicates R 4 Connection points with compounds;

[0116] Each R 5 and each R 6Independently, it is hydrogen, a halogroup, -CN, a -C1-C4 alkyl group optionally substituted with a halogroup, an -O-(C1-C4 alkyl group optionally substituted with a halogroup, an -O-(C1-C4 alkyl group substituted with a C3-C6 cycloalkyl group, an N-linked saturated 3-7 membered heterocyclic group, a 5-6 membered heteroaryl group, or a phenyl group, wherein there are no more than two R groups. 6 It is not hydrogen; the R on the adjacent ring atoms 5 and R 6 Optionally together form with R 4 Fused 4-7 membered saturated heterocycles or cycloalkyl rings;

[0117] R 7 It is hydrogen, -C1-C4 alkyl, -C1-C4 alkylene, -O-C1-C4 alkyl, -C(O)-C1-C4 alkyl, or C3-C6 cycloalkyl, wherein R 7 Any C1-C4 alkyl or C1-C4 alkylene moiety may optionally be substituted by one or more substituents independently selected from halogen and -CN;

[0118] R 8 It is hydrogen, -C1-C4 alkyl, or 4-6 membered saturated heterocycle;

[0119] R 9 It is hydrogen, a halogroup, -CN, a -C1-C4 alkyl group optionally substituted with a halogroup, an -O-C1-C4 alkyl group optionally substituted with a halogroup, or a phenyl group optionally substituted;

[0120] m is 0, 1, 2, 3 or 4;

[0121] n is 0, 1, 2, or 3; and

[0122] m+n is 1, 2, 3, or 4.

[0123] Any hydrogen atom may be optionally replaced by deuterium.

[0124] In some embodiments, this disclosure provides compounds having structural formula A:

[0125] (A),

[0126] Or its solvates, enantiomers, tautomers or diastereomers, or pharmaceutically acceptable salts of any of the foregoing substances, wherein:

[0127] X, Y, and Z are each independently CH or N;

[0128] R 1 It is -O-, -NH-, or -N(C1-C3 alkyl)-;

[0129] Each R 2Independently, it is a fluorinated group, -CN, unsubstituted C1-C5 alkyl, fluorinated C1-C5 alkyl, or cyano-substituted C1-C5 alkyl, wherein the two R groups bonded to the same carbon atom are... 2 Optionally, they can be combined to form a spirocyclic fused C3-C7 cycloalkyl ring, or two R atoms bonded to different carbon atoms. 2 Optionally, they can be combined to form a bridged C3-C5 cycloalkyl ring, or an R 2 and R 8 They can be arbitrarily combined to form bridging 3-5 element rings;

[0130] R 3 It is a -C1-C3 alkyl group or -CH2-CH=CH2;

[0131] R 4 yes , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or ,in" " indicates R 4 Connection points with compounds;

[0132] Each R 5 and each R 6 Independently, it is hydrogen, a halogroup, -CN, a -C1-C4 alkyl group optionally substituted with a halogroup, an -O-(C1-C4 alkyl group optionally substituted with a halogroup, an -O-(C1-C4 alkyl group substituted with a C3-C6 cycloalkyl group, an N-linked saturated 3-7 membered heterocyclic group, a 5-6 membered heteroaryl group, or a phenyl group, wherein there are no more than two R groups. 6 It is not hydrogen; the R on the adjacent ring atoms 5 and R 6 Optionally together form with R 4 Fused 4-7 membered saturated heterocycles or cycloalkyl rings;

[0133] R 7 It is hydrogen, -C1-C4 alkyl, -C1-C4 alkylene, -O-C1-C4 alkyl, -C(O)-C1-C4 alkyl, or C3-C6 cycloalkyl, wherein R 7 Any C1-C4 alkyl or C1-C4 alkylene moiety may optionally be substituted by one or more substituents independently selected from halogen and -CN;

[0134] R 8 It is hydrogen, -C1-C4 alkyl, or 4-6 membered saturated heterocycle;

[0135] R 9 It is hydrogen, a halogroup, -CN, a -C1-C4 alkyl group optionally substituted with a halogroup, an -O-C1-C4 alkyl group optionally substituted with a halogroup, or a phenyl group optionally substituted;

[0136] m is 0, 1, 2, or 3;

[0137] n is 0, 1, 2, or 3; and

[0138] m+n = 1, 2, 3 or 4

[0139] In formula A, any hydrogen atom may be optionally replaced by deuterium.

[0140] In some embodiments, this disclosure provides compounds having structural formula I:

[0141] (I),

[0142] Or its solvates, enantiomers, tautomers or diastereomers, or pharmaceutically acceptable salts of any of the foregoing substances, wherein:

[0143] X, Y, and Z are each independently CH or N;

[0144] R 1 It is -O-, -NH-, or -N(C1-C3 alkyl)-;

[0145] Each R 2 Independently, it is a fluorinated group, -CN, unsubstituted C1-C5 alkyl, fluorinated C1-C5 alkyl, or cyano-substituted C1-C5 alkyl, wherein the two R groups bonded to the same carbon atom are... 2 Optionally, they can be combined to form a spirocyclic fused C3-C7 cycloalkyl ring, or two R atoms bonded to different carbon atoms. 2 Optionally, they can be combined to form a bridged C3-C5 cycloalkyl ring, or an R 2 and R 8 They can be arbitrarily combined to form bridging 3-5 element rings;

[0146] R 3 It is a -C1-C3 alkyl group or -CH2-CH=CH2;

[0147] R 4 yes , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or ,in" " indicates R 4 Connection points with compounds;

[0148] Each R 5 and each R 6 Independently, it is hydrogen, a halogroup, -CN, a -C1-C4 alkyl group optionally substituted with a halogroup, or an -O-(C1-C4 alkyl group optionally substituted with a halogroup), wherein there are no more than two R groups. 6 It is not hydrogen; the R on the adjacent ring atoms 5 and R 6 Optionally together form with R 4 Fused 4-7 membered saturated heterocycles or cycloalkyl rings;

[0149] R 7 It is hydrogen, -C1-C4 alkyl or -C1-C4 alkylene-O-C1-C4 alkyl, wherein R 7 Any C1-C4 alkyl or C1-C4 alkylene moiety may optionally be substituted by one or more substituents independently selected from halogen and -CN;

[0150] R 8 It is hydrogen or -C1-C4 alkyl;

[0151] R 9 It is hydrogen, a halogroup, -CN, a -C1-C4 alkyl group optionally substituted with a halogroup, an -O-C1-C4 alkyl group optionally substituted with a halogroup, or a phenyl group optionally substituted;

[0152] m is 0 or 1;

[0153] n is 0, 1, 2, or 3; and

[0154] m and n are not both 0.

[0155] As defined and discussed above, X, Y, and Z are each independently CH or N.

[0156] In some embodiments, X, Y, and Z are each CH (i.e., the ring comprising X, Y, and Z is phenyl-2,6-diyl). In some embodiments, X is N and Y and Z are each CH (i.e., the ring is pyridin-2,6-diyl). In some embodiments, X and Y are N and Z is CH (i.e., the ring is pyrimidine-2,6-diyl). In some embodiments, X and Z are each N and Y is CH (i.e., the ring is pyrazin-2,6-diyl).

[0157] As defined and discussed throughout the text, R 1 It is -O-, -NH- or -N(C1-C3 alkyl)-.

[0158] In some implementation schemes, R 1 Yes -O-. In some implementations, R 1 It is -NH-. In some implementations, R 1 It is -N(CH3)-. In some implementations, R 1 It is -N(CH2CH3)-. In some implementations, R 1 It is -N(CH2CH2CH3)-.

[0159] As defined above and discussed throughout, each R 2 (If present) independently a fluorinated group, -CN, an unsubstituted C1-C5 alkyl group, a fluorinated C1-C5 alkyl group, or a cyano-substituted C1-C5 alkyl group, wherein the two R groups bonded to the same carbon atom are... 2 Optionally, they can be combined to form a spirocyclic fused C3-C7 cycloalkyl ring, or two R atoms bonded to different carbon atoms. 2 Optionally, they can be combined to form a bridged or fused C3-C5 cycloalkyl ring, or an R 2 and R 8 They can be arbitrarily combined to form bridging or fused 3- to 5-membered rings.

[0160] In R 2 In the definition, a fused, fused, or bridged helical ring refers to a ring formed by two R... 2 or R 2 and R 8 A ring formed together with substituents, and the ring is as follows Equation I describes the connection method of the rings. Through two R... 2 Or an R 2 And an R 8 Examples of such helical fused, fused, and bridged ring systems formed together include, but are not limited to: , , , , , and .

[0161] In some implementation schemes, R 2 It does not exist. In some implementations, an R... 2 and R 8 Together with the rings they bond to form In some implementations, an R 2 and R 8 Together with the rings they bond to form In some implementations, an R 2 and R8 Together with the rings they bond to form .

[0162] As defined and discussed throughout the text, R 3 It is a -C1-C3 alkyl group or -CH2-CH=CH2. In some embodiments, R 3 It is -CH2-CH=CH2. In some implementations, R 3 It is methyl. In some embodiments, R 3 It is ethyl. In some embodiments, R 3 It's n-propyl.

[0163] As defined and discussed throughout the text, R 4 yes , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or .

[0164] In some implementation schemes, R 4 yes In some aspects of this implementation plan, R 5 It is hydrogen. In some aspects of any of these implementation schemes, R 5 It is a C1-C4 alkyl group. In some aspects of this embodiment, R... 5 It is methyl. In some aspects of this embodiment, R 5 It is CN. In some aspects of any of these implementation schemes, R 5 It is a halogenated group. In some aspects of this implementation scheme, R... 5 It is a halogenated group. In some aspects of this implementation scheme, R... 5 It is a fluorinated group. In some aspects of this embodiment, R... 5 It is a brominated group. In some aspects of any of these embodiments, R 5 It is optionally -O-(C1-C4 alkyl) substituted with a halogroup. In some aspects of any of these embodiments, R 5 It is optionally a -O-(C1-C4 alkyl) substituted with a fluorinated group. In some aspects of this embodiment, R 5 Yes -OCF3. In some aspects of this implementation scheme, each R 6 It is hydrogen. In some aspects of this implementation scheme, an R 6 It is hydrogen and another R 6 It is a halogenated group. In some aspects of this implementation, an R 6 It is hydrogen and another R 6 It is a chlorinated group. In some aspects of this implementation, an R 6 It is hydrogen and another R 6 It is a fluorinated group. In some aspects of this embodiment, an R 6 It is hydrogen and another R 6 It is a brominated group. In some aspects of this embodiment, an R 6 It is hydrogen and another R 6 It is an -C1-C4 alkyl group optionally substituted with a halogroup. In some aspects of this embodiment, an R 6 It is hydrogen and another R 6 It is an -C1-C4 alkyl group optionally substituted with a fluorinated group. In some aspects of this embodiment, an R 6 It is hydrogen and another R 6 Yes -CH3. In some aspects of this implementation scheme, an R 6 It is hydrogen and another R 6 It is -CF3. In some aspects of this implementation scheme, an R 6It is hydrogen and another R 6 It is optionally substituted with a -O (C1-C4 alkyl group). In some aspects of this embodiment, an R 6 It is hydrogen and another R 6 It is optionally substituted with a fluorinated group (-O, C1-C4 alkyl). In some aspects of this embodiment, an R 6 It is hydrogen and another R 6 It is -OCH3. In some aspects of this implementation scheme, an R 6 It is hydrogen and another R 6 It is -OCH2CF3. In some aspects of this implementation, an R 6 It is hydrogen and another R 6 It is -CN. In some aspects of this implementation, R is bonded to adjacent ring atoms. 5 and R 6 Together they form a methylenedioxy group. In some aspects of this embodiment, an R 6 It is hydrogen and another R 6 It is cyclopropylmethoxy. In some aspects of this embodiment, an R 6 It is hydrogen and another R 6 It is a morpholino group. In some aspects of this embodiment, an R 6 It is methyl and another R 6 It is a morpholino group. In some aspects of this embodiment, an R 6 It is hydrogen and another R 6 It is a thiomorpholino group. In some aspects of this embodiment, an R 6 It is hydrogen and another R 6 It is 2-methylpropyl-1-yloxy. In some aspects of this embodiment, an R 6 It is hydrogen and another R 6 It is 1-methyl-1H-pyrazolyl. In some aspects of this embodiment, an R 6 It is hydrogen and another R 6 It is a phenyl group optionally substituted with a halogroup. In some aspects of this embodiment, an R 6 It is hydrogen and another R 6 It is an ethoxy group that is optionally substituted with a halogenated group.

[0165] In some implementation schemes, R 4It is any one of phenyl, 3-methylphenyl, 3-chloro-5-bromophenyl, 3,4-dichlorophenyl, 4-bromophenyl, 4-chlorophenyl, 4-fluorophenyl, 4-(1-methylpiperidin-4-yl)-5-methylphenyl, 4-trifluoromethoxyphenyl, 3-methyl-4-trifluoromethoxyphenyl, 4-(2,2,2-trifluoroeth-1-yl)oxyphenyl, 3-methyl-4-(2,2,2-trifluoroeth-1-yl)oxyphenyl, 4-trifluoromethylphenyl, 3-methyl-4-chlorophenyl, 3-methyl-4-fluorophenyl, 3-methyl-5-fluorophenyl, 3-methyl-4-cyanophenyl, 3-methyl-4-methoxyphenyl, 3-methyl-4-(2,2,2-trifluoroeth-1-yloxy)phenyl, or 3,4-methylenedioxyphenyl.

[0166] In some implementation schemes, R 4 It is 4-(cyclopropylmethyloxy)phenyl, 4-(morpholin-4-yl)phenyl, 4-(thiomorpholin-4-yl)phenyl, 4-(2-methylpropyl-1-yloxy)phenyl, 3-(1-methyl-1H-pyrazol-4-yl)phenyl, 4-(1-methyl-1H-pyrazol-4-yl)phenyl, 3-methyl-4-(morpholin-4-yl)phenyl, 4-(phenyl)phenyl, 3-bromophenyl, 3-cyanophenyl, 4 Any one of -(2-fluoroethoxy-1-yl)phenyl, 4-cyanophenyl, 4-(1,1-dioxothiazin-4-yl)phenyl, 3-(pyridin-3-yl)phenyl, 3-(pyrimidin-5-yl)phenyl, 3-(1H-imidazol-1-yl)phenyl, 3-(1H-pyrazol-1-yl)phenyl, 3-(d3-methyl)phenyl, 4-(3-fluoroprop-1-yl)phenyl or 4-ethoxyphenyl.

[0167] In some implementation schemes, R 4 yes In some implementations, R 4 yes In some aspects of either of these implementation schemes, R 5 It is hydrogen. In some aspects of any of these implementation schemes, R 5 It is a halogenated group. In some aspects of any of these implementations, R 5 It is a chlorinated group. In some aspects of any of these embodiments, R 5 It is a fluorinated group. In some aspects of any of these embodiments, R 5 It is cyano. In some aspects of any of these embodiments, R 5 It is a C1-C4 alkyl group optionally substituted with a halogroup. In some aspects of any of these embodiments, R 5 It is a C1-C4 alkyl group optionally substituted with a fluorinated group. In some aspects of any of these embodiments, R5 It is an -O-C1-C4 alkyl group. In some aspects of any of these embodiments, R 5 Methoxylated. In some aspects of any of these embodiments, R 5 It is methyl. In some aspects of any of these embodiments, R 5 Yes -CF3. In some aspects of any of these implementations, R 6 It is hydrogen. In some aspects of any of these implementation schemes, R 6 It is methyl. In some aspects of any of these embodiments, R 6 It is an -O-C1-C4 alkyl group. In some aspects of any of these embodiments, R 6 It is methoxylated. In some aspects of any of these embodiments, R 6 It is an -O-C1-C4 haloalkyl group. In some aspects of any of these embodiments, R 6 It is 2-fluoroethoxy.

[0168] In some implementation schemes, R 4 It is any one of 2-methylpyridin-4-yl, 6-methylpyridin-3-yl, 2,6-dimethylpyridin-4-yl, 2-trifluoromethyl-6-methylpyridin-4-yl or 2-trifluoromethylpyridin-4-yl.

[0169] In some implementation schemes, R 4 It is any one of pyridin-3-yl, 2-methylpyridin-5-yl, 2-methoxypyridin-4-yl, 2-methoxypyridin-5-yl, 3-chloropyridin-5-yl, 3-fluoropyridin-5-yl, 3-cyanopyridin-5-yl, 3-methylpyridin-5-yl or 3-methoxypyridin-5-yl.

[0170] In some implementation schemes, R 4 It is 2-(2-fluoroethoxy)pyridin-5-yl.

[0171] In some implementation schemes, R 4 yes In some aspects of this implementation plan, R 7 It is a C1-C4 alkyl group optionally substituted with a halogroup. In some aspects of this embodiment, R 7 It is a C1-C4 alkyl group optionally substituted with a fluorinated group. In some aspects of this embodiment, R 7 It is methyl. In some aspects of this embodiment, R 7 It is ethyl. In some aspects of this embodiment, R 7 It is propyl. In some aspects of this implementation scheme, R 7 It is isopropyl. In some aspects of this implementation scheme, R...7 It is -CH2CH(CH3)2. In some aspects of this implementation scheme, R 7 It is -CH2C(CH3)2F. In some aspects of this implementation scheme, R 7 It is -CH2CH2CF3. In some aspects of this implementation scheme, R 4 It is any one of 1-methyl-1H-pyrazole-4-yl, 1-propyl-1H-pyrazole-4-yl, 1-isopropyl-1H-pyrazole-4-yl, 1-(2,2-dimethylethyl-1-yl)pyrazole-4-yl, 1-(2-fluoro-2,2-dimethylethyl-1-yl)-1H-pyrazole-4-yl or 1-(3,3,3-trifluoropropyl-1-yl)-1H-pyrazole-4-yl.

[0172] In some implementation schemes, R 4 yes In some aspects of this implementation scheme, each R 6 It is hydrogen. In some aspects of this implementation scheme, R 7 It is hydrogen. In some aspects of this implementation scheme, R 7 It is a C1-C4 alkyl group optionally substituted with a halogroup. In some aspects of this embodiment, R 7 It is a C1-C4 alkyl group optionally substituted with a fluorinated group. In some aspects of this embodiment, R 7 It is methyl. In some aspects of this embodiment, R 7 It is ethyl. In some aspects of this embodiment, R 7 It is propyl. In some aspects of this implementation scheme, R 7 It is isopropyl. In some aspects of this implementation scheme, R... 7 It is -CH2CH(CH3)2. In some aspects of this implementation scheme, R 7 It is -CH2C(CH3)2F. In some aspects of this implementation scheme, R 7 It is -CH2CH2CF3. In some aspects of this implementation scheme, R 7 It is cyclopropyl. In some aspects of this implementation scheme, R 7 It is isobutyl. In some aspects of this implementation, R... 7 It is 3-fluoropropyl. In some aspects of this embodiment, R 4 It is 1-methyl-1H-indazole-5-yl. In some embodiments, R 4It is selected from 1H-indazole-5-yl, 1-methyl-1H-indazole-5-yl, 1-ethyl-1H-indazole-5-yl, 1-isopropyl-1H-indazole-5-yl, 1-propyl-1H-indazole-5-yl, 1-(3-fluoroprop-1-yl)-1H-indazole-5-yl, 1-isobutyl-1H-indazole-5-yl and 1-cyclopropyl-1H-indazole-5-yl.

[0173] In some implementations, when R 4 yes At that time, an R 6 It is hydrogen and another R 6 It is a methyl or halogroup. In some aspects of this embodiment, R 4 It is 3-chloro-1H-indazole-5-yl, 3-methyl-1H-indazole-5-yl, or 1,3-dimethyl-1H-indazole-5-yl.

[0174] In some implementation schemes, R 4 yes In some aspects of this implementation plan, R 7 It is hydrogen. In some aspects of this implementation scheme, R 7 It is methyl. In some aspects of this embodiment, R 7 It is ethyl. In some aspects of this embodiment, R 7 It is tert-butyl. In some aspects of this implementation scheme, each R 6 It is hydrogen. In some aspects of this implementation scheme, R 4 It is 2-methyl-2H-indazole-5-yl or 2-ethyl-2H-indazole-5-yl. In some aspects of this embodiment, R 4 It is 2-(tert-butyl)-2H-indazole-5-yl.

[0175] In some implementation schemes, R 4 yes In some aspects of this implementation plan, R 9 It is an optionally substituted phenyl group. In some aspects of this embodiment, R 9 It is an unsubstituted phenyl group. In some aspects of this embodiment, R... 4 It is 3-phenylisothiazolyl-5-yl.

[0176] In some implementation schemes, R 4 yes In some implementations, R 4 yes In some aspects of each of these implementation schemes, each R 6 It is hydrogen. In some aspects of each of these implementation schemes, the two Rs 6 It is hydrogen and another R 6It is not hydrogen. In some aspects of each of these implementation schemes, an R 6 It is hydrogen and the other two R 6 It is not hydrogen. In some aspects of this implementation scheme, R... 4 It is benzofuran-6-yl. In some aspects of this embodiment, R 4 It is benzofuran-5-yl.

[0177] In some implementation schemes, R 4 yes In some aspects of this implementation plan, R 7 It is a C1-C4 alkyl group. In some aspects of this embodiment, R... 7 It is methyl. In some aspects of this embodiment, R 7 It is -C(O)CH3. In some aspects of this implementation scheme, each R 6 It is hydrogen. In some aspects of this implementation scheme, the two Rs 6 It is hydrogen and another R 6 It is not hydrogen. In some aspects of this implementation scheme, an R 6 It is hydrogen and the other two R 6 It is not hydrogen. In some aspects of this implementation scheme, R... 4 It is 1-methyl-1H-indole-5-yl. In some aspects of this embodiment, R 4 It is 1-acetyl-1H-indole-5-yl.

[0178] In some implementation schemes, R 4 yes In some aspects of this implementation scheme, each R 6 It is hydrogen. In some aspects of this implementation scheme, R 4 It is 1,2,4-triazolo[1,5-a]pyridin-6-yl.

[0179] In some implementation schemes, R 4 yes In some aspects of this implementation scheme, each R 6 It is hydrogen. In some aspects of this implementation scheme, R 4 It is quinoxaline-6-yl.

[0180] In some implementation schemes, R 4 Selected from , , and Any one of them. In some aspects of this implementation scheme, each R 6 It is hydrogen. In some aspects of this implementation scheme, R 4 It is quinoline-6-yl, quinoline-7-yl, isoquinoline-6-yl, or isoquinoline-7-yl.

[0181] In some implementation schemes, R 4 yes or In some aspects of these implementation schemes, each R 6 It is hydrogen. In some aspects of these implementation schemes, an R 6 It is hydrogen and another R 6 It is methyl. In some aspects of these embodiments, R 4 It is 2-methylbenzo[d]oxazol-5-yl or 2-methylbenzo[d]oxazol-6-yl.

[0182] In some implementation schemes, R 4 yes In some aspects of these implementation schemes, each R 6 It is hydrogen. In some aspects of these implementation schemes, R 4 It is benzo[d]isoxazol-6-yl.

[0183] In some implementation schemes, R 4 yes In some aspects of this implementation scheme, each R 6 It is hydrogen. In some aspects of this implementation scheme, an R 6 It is hydrogen and another R 6 It is methyl. In some aspects of this embodiment, R 4 It is benzo[d]thiazol-6-yl or 2-methylbenzo[d]thiazol-6-yl.

[0184] In some implementation schemes, R 4 yes In some aspects of this implementation scheme, each R 6 It is hydrogen. In some aspects of this implementation scheme, R 7 It is methyl. In some aspects of this embodiment, R 4 It is 1-methyl-1H-benzo[d]imidazol-5-yl. In some aspects of this embodiment, R 7 It is isopropyl. In some aspects of this implementation scheme, R... 4 It is 1-isopropyl-1H-benzo[d]imidazol-5-yl.

[0185] In some implementation schemes, R 4 yes In some aspects of this implementation scheme, each R 6 It is hydrogen. In some aspects of this implementation scheme, R 4 It is benzo[d]thiazolyl-5-yl.

[0186] In some implementation schemes, R 4 yes In some aspects of this implementation scheme, each R 6 It is hydrogen. In some aspects of this implementation scheme, R 7 It is methyl. In some aspects of this embodiment, R 4 It is 1-methyl-1H-indazole-6-yl.

[0187] In some implementation schemes, R 4 yes In some aspects of this implementation scheme, each R 6 It is hydrogen. In some aspects of this implementation scheme, R 7 It is methyl. In some aspects of this embodiment, R 4 It is 2-methyl-2H-indazole-6-yl.

[0188] In some implementation schemes, R 4 yes In some aspects of this implementation scheme, each R 6 It is hydrogen. In some aspects of this implementation scheme, R 4 It is imidazo[1,2-a]pyridin-6-yl.

[0189] In some implementation schemes, R 4 yes In some aspects of this implementation plan, R 6 It is hydrogen. In some aspects of this implementation scheme, R 7 It is a C1-C4 alkyl group. In some aspects of this embodiment, R... 7 It is isopropyl. In some aspects of this implementation scheme, R... 4 It is 1-isopropyl-1H-pyrazolo[3,4-b]pyridin-5-yl.

[0190] In some implementation schemes, R 4 yes In some aspects of this implementation plan, R 6 It is hydrogen. In some aspects of this implementation scheme, R 7 It is a C1-C4 alkyl group. In some aspects of this embodiment, R... 7 It is isopropyl. In some aspects of this implementation scheme, R... 4 It is 1-isopropyl-1H-benzo[d][1,2,3]triazol-5-yl.

[0191] As defined and discussed throughout the text, R 5 It is hydrogen, a halogroup, -CN, a -C1-C4 alkyl group optionally substituted with a halogroup, or an -O-(C1-C4 alkyl group optionally substituted with a halogroup, or an R on an adjacent ring atom. 5 and R 6 Together with R4 Fused 4-7 membered saturated heterocyclic or cycloalkyl rings. In some embodiments, R 5 It is hydrogen. In some implementations, R 5 It is a halogenated group. In some implementations, R 5 It is a chlorinated group. In some implementations, R 5 It is a fluorinated group. In some implementations, R 5 It is a C1-C4 alkyl group optionally substituted with a halogroup. In some embodiments, R 5 It is a C1-C4 alkyl group optionally substituted with a fluorinated group. In some embodiments, R 5 It is methyl. In some embodiments, R 5 It is -CF3. In some implementations, R on adjacent ring atoms 5 and R 6 Together with R 4 Fused 4-7 membered saturated heterocycles or cycloalkyl rings. In some embodiments, R on adjacent ring atoms 5 and R 6 Together with R 4 Fused methylenedioxy groups.

[0192] As defined above and discussed throughout, each R 6 Independently, it is hydrogen, a halogroup, -CN, a -C1-C4 alkyl group optionally substituted with a halogroup, or an -O-(C1-C4 alkyl group optionally substituted with a halogroup), wherein there are no more than two R groups. 6 It is not hydrogen. In some implementations, no more than one R 6 It is not hydrogen. In some implementations, each R 6 It is hydrogen. In some implementations, an R 6 It is a halogenated group. In some implementations, an R 6 It is a chlorinated group. In some implementations, an R 6 It is a fluorinated group. In some embodiments, an R 6 It is a brominated group. In some embodiments, an R 6 It is an -C1-C4 alkyl group optionally substituted with a halogroup. In some embodiments, an R 6 It is an -C1-C4 alkyl group optionally substituted with a fluorinated group. In some embodiments, an R 6 It is -CH3. In some implementations, an R 6 It is -CF3. In some aspects of this implementation scheme, an R 6 It is hydrogen and another R 6 It is a halogenated group. In some implementations, an R 6It is optionally substituted with a -O (C1-C4 alkyl group). In some embodiments, an R 6 It is optionally substituted with a fluorinated group (-O, C1-C4 alkyl). In some embodiments, an R 6 It is -OCH3. In some implementations, an R 6 It is -OCH2CF3. In some implementations, an R 6 Yes - CN.

[0193] As defined and discussed throughout the text, R 7 It is hydrogen, -C1-C4 alkyl, -C1-C4 alkylene, -O-C1-C4 alkyl, -C(O)-C1-C4 alkyl, or C3-C6 cycloalkyl, wherein R 7 Any C1-C4 alkyl or C1-C4 alkylene moiety is optionally substituted by one or more substituents independently selected from halogenated and -CN groups. In some embodiments, R 7 It is a C1-C4 alkyl group optionally substituted with a halogroup. In some embodiments, R 7 It is a C1-C4 alkyl group optionally substituted with a fluorinated group. In some embodiments, R 7 It is methyl. In some embodiments, R 7 It is ethyl. In some embodiments, R 7 It is propyl. In some implementations, R 7 It is isopropyl. In some implementations, R 7 It is -CH2CH(CH3)2. In some implementations, R 7 It is -CH2C(CH3)2F. In some implementations, R 7 It is -CH2CH2CF3. In some implementations, R 7 It is -CH2CH2CH2F3. In some implementations, R 7 It is cyclopropyl. In some implementations, R 7 It is -C(O)CH3.

[0194] As defined and discussed throughout the text, R 8 It is hydrogen, -C1-C4 alkyl, or a 4-6 membered saturated heterocycle. In some embodiments, R 8 It is hydrogen. In some implementations, R 8 It is methyl. In some embodiments, R 8 It is ethyl. In some embodiments, R 8 It is propyl. In some implementations, R 8 It is oxocyclic butyl-3-yl.

[0195] As defined and discussed throughout the text, R9 It is hydrogen, a halogroup, -CN, a -C1-C4 alkyl group optionally substituted with a halogroup, an -O-C1-C4 alkyl group optionally substituted with a halogroup, or an optionally substituted phenyl group. In some embodiments, R 9 It is an unsubstituted phenyl group.

[0196] As defined and discussed throughout the text, the structure is as follows: The monocyclic heterocycle represented can be saturated with between 4 (when the sum of m+n is 1) and 7 (when the sum of m+n is 4) ring atoms. In some embodiments, when two R atoms bonded to the same carbon atom... 2 When they together form a C3-C7 cycloalkyl ring, the ring can be a spirocyclic fused bicyclic ring. In some embodiments, when two R atoms are bonded to different carbon atoms... 2 When they together form a C3-C5 cycloalkyl ring, the ring can be a bridged bicyclic ring. In some embodiments, when an R 2 and R 8 When they form a 3-5 membered ring together, the ring can be a bridged bicyclic ring. In some embodiments, when two R atoms bonded to different carbon atoms... 2 When they together form a C3-C5 cycloalkyl ring, the ring can be a fused bicyclic ring. In some embodiments, when an R 2 and R 8 When they form a 3-5 member ring, the ring can be a fused bicyclic ring.

[0197] In some implementation schemes, the structure is as follows: The ring represented is a piperidinyl ring. In some aspects of these embodiments, the piperidinyl ring is a piperidin-3-yl ring (i.e., n is 1 and m is 2; or n is 3 and m is 0). In some aspects of these embodiments, the piperidinyl ring is a piperidin-4-yl ring (yl, n is 2 and m is 1). In some aspects of these embodiments, the R on such a piperidinyl ring... 8 The substituent is hydrogen. In some aspects of these embodiments, the R on such a piperidinyl ring... 8 The substituent is methyl. In some aspects of these embodiments, the R on such a piperidinyl ring... 8 The substituent is a trideuterated methyl group (methyl-d3). In some aspects of these embodiments, the R on such a piperidinyl ring... 8 The substituent is ethyl. In some aspects of these embodiments, the R on such a piperidinyl ring... 8 The substituent is isopropyl. In some aspects of these embodiments, the R on such a piperidinyl ring... 8 The substituent is propyl. In some aspects of these embodiments, R 2 It does not exist. In some aspects of these implementation schemes, one or more R 2It is methyl. In some aspects of these embodiments, the piperidinyl ring is piperidinyl-4-yl, piperidinyl-3-yl, 1-methylpiperidin-4-yl, 1-d3-methylpiperidin-4-yl, 1-methylpiperidin-3-yl, 1-ethylpiperidin-4-yl, 1-propylpiperidin-4-yl, 1,2-dimethylpiperidin-4-yl, 2,2-dimethylpiperidin-4-yl, 2-methylpiperidin-4-yl, 1,2,2-trimethylpiperidin-4-yl, or 1-(oxecyclobut-3-yl)piperidin-4-yl.

[0198] In some implementation schemes, the structure is as follows: The ring represented is a pyrrolidinyl ring. In some aspects of these embodiments, the pyrrolidinyl ring is a pyrrolidin-3-yl ring (i.e., n is 1 and m is 1; or n is 2 and m is 0). In some aspects of these embodiments, the R on such a pyrrolidinyl ring... 8 The substituent is hydrogen. In some aspects of these embodiments, the R on such a pyrroleyl ring... 8 The substituent is methyl. In some aspects of these embodiments, the R on such a pyrrolidinyl ring... 8 The substituent is ethyl. In some aspects of these embodiments, the R on such a pyrrolidinyl ring... 8 The substituent is isopropyl. In some aspects of these embodiments, R 2 It does not exist.

[0199] In some implementation schemes, the structure is as follows: The ring represented is a quinine-based ring. In some aspects of these embodiments, the quinine-based ring is a quinine-3-based ring (i.e., n is 1 and m is 2; or n is 3 and m is 0; and an R 2 and R 8 Together they form a bridging 4-membered ring. In some aspects of these implementations, R on such a quinine-based ring... 8 The substituent is hydrogen. In some aspects of these embodiments, the R on such a quinine ring is... 8 The substituent is methyl. In some aspects of these embodiments, the R on such a quinine ring is... 8 The substituent is ethyl. In some aspects of these embodiments, the R on such a quinine ring is... 8 The substituent is isopropyl. In some aspects of these embodiments, R 2 It does not exist.

[0200] In some implementation schemes, the structure is as follows: The ring represented is an aziridine [3.2.1]octyl ring. In some aspects of these embodiments, the aziridine [3.2.1]octyl ring is an 8-aziridine [3.2.1]octane-3-yl ring (i.e., n is 1 and m is 2; or n is 3 and m is 0; and two R atoms are bonded to different carbon atoms). 2 Together they form a bridging C4 cycloalkyl ring. In some aspects of these embodiments, the R on such a aziridine bicyclic [3.2.1] octyl ring 8 The substituent is hydrogen. In some aspects of these embodiments, the R on the octyl ring of this type of azabicyclic [3.2.1] 8 The substituent is methyl. In some aspects of these embodiments, the R on the octyl ring of this type of azabicyclic [3.2.1] 8 The substituent is ethyl. In some aspects of these embodiments, the R on the octyl ring of this type of azabicyclic [3.2.1] 8 The substituent is isopropyl. In some aspects of these embodiments, R 2 It does not exist.

[0201] In some implementation schemes, the structure is as follows: The ring represented is an aziridine bicyclic [3.3.1]nonyl ring. In some aspects of these embodiments, the aziridine bicyclic [3.3.1]nonyl ring is a 9-aziridine bicyclic [3.3.1]nonane-3-yl ring (i.e., n is 1 and m is 2; or n is 3 and m is 0; and two R atoms are bonded to different carbon atoms). 2 Together they form a bridging C5 cycloalkyl ring. In some aspects of these embodiments, the R on such a aziridine bicyclic [3.3.1]nonyl ring 8 The substituent is hydrogen. In some aspects of these embodiments, the R on the nonyl ring of this type of azabicyclic [3.3.1] 8 The substituent is methyl. In some aspects of these embodiments, the R on the nonyl ring of this type of azabicyclic [3.3.1] 8 The substituent is ethyl. In some aspects of these embodiments, the R on the nonyl ring of this type of azabicyclic [3.3.1] 8 The substituent is isopropyl. In some aspects of these embodiments, R 2 It does not exist.

[0202] In some implementation schemes, the structure is as follows: The ring represented is an aziridine [2.2.2]octyl ring. In some aspects of these embodiments, the aziridine [2.2.2]octyl ring is a 2-aziridine [2.2.2]octane-5-yl ring (i.e., n is 1 and m is 2; or n is 3 and m is 0; and two R atoms are bonded to different carbon atoms). 2Together they form a bridging C4 cycloalkyl ring. In some aspects of these embodiments, the R on such a aziridine bicyclic [2.2.2] octyl ring 8 The substituent is methyl.

[0203] In some implementation schemes, the structure is as follows: The ring represented is an azircyclic heptyl ring. In some aspects of these embodiments, the azircyclic heptyl ring is an azircyclic heptane-4-yl (i.e., n is 2 and m is 2). In some aspects of these embodiments, the R on such an azircyclic heptyl ring... 8 The substituent is hydrogen. In some aspects of these embodiments, the R on such a nitrogen-containing heptyl ring... 8 The substituent is methyl.

[0204] In some embodiments, the compound of Formula I may be a compound selected from those in Table 1 or a pharmaceutically acceptable salt thereof, which lists the chemical structure of each compound and the LC-MS and analytical results of the compounds analyzed therein. 1 ¹H NMR. Chemical shifts are reported as ppm (δ) using residual solvent as an internal standard. Peak multiplicity given in Hz is as follows: s, singlet; d, doublet; dd, double doublet; ddd, double doublet; t, triplet; dt, double triplet; q, quartet; dq, double quartet; p, quintet; h, septet; m, multiplet; br s, broad singlet.

[0205] Table 1. Exemplary Compounds

[0206]

[0207]

[0208]

[0209]

[0210]

[0211]

[0212]

[0213]

[0214]

[0215]

[0216]

[0217]

[0218]

[0219]

[0220]

[0221]

[0222]

[0223]

[0224]

[0225]

[0226]

[0227]

[0228]

[0229]

[0230]

[0231]

[0232]

[0233]

[0234]

[0235]

[0236]

[0237]

[0238]

[0239]

[0240]

[0241]

[0242]

[0243]

[0244]

[0245]

[0246]

[0247]

[0248]

[0249]

[0250]

[0251]

[0252]

[0253]

[0254]

[0255]

[0256]

[0257]

[0258]

[0259]

[0260]

[0261]

[0262]

[0263]

[0264]

[0265]

[0266]

[0267]

[0268]

[0269]

[0270]

[0271]

[0272]

[0273]

[0274]

[0275]

[0276]

[0277]

[0278]

[0279]

[0280]

[0281]

[0282]

[0283]

[0284]

[0285]

[0286]

[0287] Or its pharmaceutically acceptable salt.

[0288] 4. Uses, formulations and application

[0289] Pharmaceutically acceptable compositions

[0290] According to another embodiment, this disclosure provides compositions comprising a compound of the present disclosure or a pharmaceutically acceptable derivative thereof, and a pharmaceutically acceptable carrier, adjuvant, or mediator. The amount of the compound in the compositions of the present disclosure is such that it effectively and measurably inhibits Wee1A kinase or a mutant thereof in a biological sample or patient. In some embodiments, the compositions of the present disclosure are formulated for administration to a patient requiring such a composition. In some embodiments, the compositions of the present disclosure are formulated for oral administration to a patient.

[0291] As used in this article, the term "patient" refers to an animal, preferably a mammal, and most preferably a human.

[0292] The term "pharmaceutically acceptable carrier, adjuvant, or mediator" refers to a non-toxic carrier, adjuvant, or mediator that does not impair the pharmacological activity of the compound formulated therewith. Pharmaceutically acceptable carriers, adjuvants, or mediators that may be used in the compositions disclosed herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffering substances such as phosphates, glycine, sorbic acid, potassium sorbate, mixtures of partial glycerides of saturated vegetable fatty acids, water, salts, or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, silica gel, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polypropylene block polymers, polyethylene glycol, and lanolin.

[0293] "Pharmaceutically acceptable derivative" means any non-toxic salt, ester, salt of ester or other derivative of the compound disclosed herein, which, when administered to a recipient, can directly or indirectly provide the compound disclosed herein or its active metabolites or residues.

[0294] The compositions disclosed herein can be administered orally, parenterally, via inhaled aerosol, topically, rectally, nasally, buccally, vaginally, or via an implanted reservoir. As used herein, the term "parenteral" includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrasheathic, intrahepatic, intralesional, and intracranial injection or infusion techniques. Preferably, the compositions are administered orally, intraperitoneally, or intravenously. The sterile injectable form of the compositions disclosed herein may be an aqueous or oily suspension. These suspensions may be formulated using suitable dispersants or wetting agents and suspending agents according to techniques known in the art. The sterile injectable formulation may also be a sterile injectable solution or suspension in a non-toxic parenteral diluent or solvent, such as a solution in 1,3-butanediol. Among acceptable media and solvents, water, Ringer's solution, and isotonic sodium chloride solution may be used. In addition, sterile non-volatile oils are routinely used as solvents or suspension media.

[0295] To achieve this purpose, any mild, non-volatile oil can be used, including synthetic monoglycerides or diglycerides. Fatty acids (such as oleic acid and its glyceride derivatives) can be used in the preparation of injections, as can natural, pharmaceutically acceptable oils such as olive oil or castor oil (especially in their polyoxyethylene forms). These oil solutions or suspensions may also contain long-chain alcohol diluents or dispersants, such as carboxymethyl cellulose or similar dispersants commonly used in the formulation of pharmaceutically acceptable dosage forms (including emulsions and suspensions). Other commonly used surfactants, such as Tween and Span, and other emulsifiers or bioavailability enhancers commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms, may also be used to achieve the desired formulation.

[0296] In some embodiments, the compounds or compositions disclosed herein are administered orally. The pharmaceutically acceptable compositions of this disclosure can be administered orally 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, common carriers include lactose and corn starch. Lubricants, such as magnesium stearate, are often also added. For oral administration in capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions are intended for oral use, the active ingredient is combined with an emulsifier and a suspending agent. Sweeteners, flavoring agents, or coloring agents may also be added if desired.

[0297] Alternatively, the pharmaceutically acceptable compositions of this disclosure can be administered in suppository form for rectal use. These can be prepared by mixing a suitable, non-irritating excipient that is solid at room temperature but liquid at rectal temperature and thus melts in the rectum to release the drug. Such materials include cocoa butter, beeswax, and polyethylene glycol.

[0298] The pharmaceutically acceptable compositions disclosed herein can also be applied topically, particularly when the therapeutic target includes areas or organs easily accessible by topical application, including diseases of the eye, skin, or lower intestine. Suitable topical formulations are readily prepared for each of these areas or organs.

[0299] Local application to the lower intestine can be achieved with rectal suppositories (see above) or with suitable enema formulations. Topical transdermal patches may also be used.

[0300] For topical application, the provided pharmaceutically acceptable compositions can be formulated into suitable ointments containing an active ingredient suspended or dissolved in one or more carriers. Carriers for the topical application of the compounds of this disclosure include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compounds, emulsified waxes, and water. Alternatively, the provided pharmaceutically acceptable compositions can be formulated into suitable lotions or creams containing an active ingredient suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, dehydrated sorbitan monostearate, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecyl alcohol, benzyl alcohol, and water.

[0301] For ophthalmic applications, the provided pharmaceutically acceptable composition can be formulated into a micronized suspension in isotonic, pH-adjusted sterile saline, or preferably, into a solution in isotonic, pH-adjusted sterile saline, with or without a preservative such as benzalkonium chloride. Alternatively, for ophthalmic applications, the pharmaceutically acceptable composition can be formulated into an ointment such as petrolatum.

[0302] The pharmaceutically acceptable compositions disclosed herein can also be administered via nasal aerosol or inhalation. Such compositions are prepared according to techniques well known in the field of pharmaceutical formulation, and can be prepared into solutions in physiological saline using benzyl alcohol or other suitable preservatives, bioavailability enhancers, fluorocarbons, and / or other conventional solubilizers or dispersants.

[0303] Most preferably, the pharmaceutically acceptable compositions of this disclosure are formulated for oral administration.

[0304] The amount of the disclosed compounds, which can be combined with a carrier substance to produce a single dosage form, will vary depending on the host being treated and the specific administration method. Preferably, the provided compositions are formulated such that an inhibitory dose of 0.001-100 mg / kg body weight / day can be administered to patients receiving these compositions.

[0305] It should also be understood that the specific dosage and treatment regimen for any particular patient will depend on a variety of factors, including the activity of the specific compound used, age, weight, health status, sex, diet, timing of administration, excretion rate, drug combination, and the judgment of the treating physician and the severity of the specific disease being treated. The amount of the disclosed compounds in the composition will also depend on the specific compounds in the composition.

[0306] Use of compounds and pharmaceutically acceptable compositions

[0307] The compounds and compositions described herein are generally used to inhibit the protein kinase activity of one or more enzymes.

[0308] Examples of kinases inhibited by the compounds and compositions described herein and to which the methods described herein are effective include Wee1A kinase, or both Wee1A kinase and Myt1 kinase.

[0309] The activity of compounds used as inhibitors of Wee1A kinase or Myt1 kinase or mutants of any of the foregoing in this disclosure can be determined in vitro, in vivo, or in cell lines. In vitro assays include determinations of the phosphorylation activity and / or subsequent functional consequences or ATPase activity of inhibitors of activated Wee1A kinase, activated Myt1 kinase, or mutants of any of the foregoing. Alternative in vitro assays quantify the ability of the inhibitor to bind to Wee1A kinase.

[0310] In recent years, inhibition of the DNA damage response (DDR) pathway has attracted considerable interest in cancer treatment, and various DDR inhibitors have been developed. Among them, the most promising inhibitors target the Wee1 kinase family, which plays a crucial role in cell cycle regulation and DNA damage recognition and repair in both non-malignant and cancer cells.

[0311] Wee1 kinase family

[0312] The Wee1 kinase family consists of three serine / threonine kinases that share a conserved molecular structure and are encoded by the following genes: WEE1 (Alternatively, WEE1 G2 checkpoint kinase or Wee1A kinase) PKMYT1 (Alternatively, MYT1 kinase or membrane-associated tyrosine and threonine-specific cdc2 inhibitory kinase) and WEE2 (Alternatively, WEE2 oocyte meiosis inhibitory kinase or Wee1B kinase). In eukaryotic somatic cells, Wee1A and Myt1 kinases play key roles in cell cycle regulation, particularly in the transition to mitosis (Schmidt M, Rohe A, Platzer C, et al., Regulation of G2 / M transition by inhibition of Wee1 and PMyt1 kinases). Molecules(2017;22:2045). Their roles as regulators are crucial in normal cell cycle progression and in responding to DNA damage as part of the DNA damage response (DDR) pathway. Similarly, Wee1B kinase regulates cell cycle progression, and particularly meiosis (Solc P, Schultz RM, Motlik J. Prophase I arrest and progression to metaphase I in mouse oocytes: Comparison of resumption of meiosis and recovery from G2-arrest in somatic cells). Mol Hum Reprod . 2010;16:654-64).

[0313] Wee1B kinase

[0314] Wee1B kinase expression is germ cell specific and inhibits meiosis by phosphorylating Tyr15 of the CDK1-cyclin B complex (JY Zhu et al., J Med Chem. 2017; 60 (18), 7863-7875). Because of its well-characterized role in meiosis, previous and current drug development efforts have not focused on Wee1B kinase. Wee1B kinase plays a dual regulatory role in oocyte meiosis by preventing premature restart before ovulation and allowing exit from metaphase II at fertilization (Nakanishi M, Ando H, Watanabe N, et al., Identification and characterization of human Wee1B, a new member of the Wee1 family of Cdk-inhibitory kinases). Genes Cells . 2000;5(10):839-47). Although identified in several cancer types WEE2 Somatic mutations (1.9% of cases) and copy number (CN) alterations (CN loss in 22.5% of patients and CN increase in 22.5% of patients) (https: / / portal.gdc.cancer.gov) have been observed, but they have not yet been functionally linked to tumor development.

[0315] Myt1 kinase

[0316] Myt1 kinase is a multifunctional protein kinase located in the ER-Golgi complex, known to play a regulatory role in the cell cycle by inhibiting Cdk1 / cyclin B1-mediated mitosis (JY Zhu et al., J Med Chem. 2017; 60 (18), 7863-7875). As described above and throughout the paper, Myt1 kinase inhibits Cdk1 / cyclin B1 activity by phosphorylating Tyr15 and Thr14 of Cdk1 and isolating Cdk1 from the nucleus. Furthermore, Myt1 kinase is also involved in the reassembly of the ER-Golgi complex at the end of mitosis.

[0317] Wee1A kinase

[0318] Wee1A kinase regulates cell entry into mitosis during the G2 / M transition in S phase by phosphorylating Tyr15 of Cdk1, thereby inactivating the Cdk1 / cyclin B complex. Cells with disrupted G1 checkpoint activity (e.g., cancer cells) depend on Wee1A kinase to inhibit Cdk1, allowing G2 / M arrest for DNA repair. If Wee1A kinase activity is altered, disrupted cells may enter mitosis prematurely without the opportunity to fully replicate their DNA content or repair potential DNA damage that may occur during S phase. This characterization of the role of Wee1A kinase in the cell cycle makes it an attractive target for anticancer therapies, especially when used in combination with DNA damage agents (JY Zhu et al., J Med Chem. 2017; 60(18), 7863-7875).

[0319] As used herein, the terms "treatment," "treat," and "treating" refer to reversing or alleviating a disease or condition as described herein, or one or more symptoms thereof, delaying its onset, or inhibiting its progression. In some embodiments, treatment may be administered after one or more symptoms have developed. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered to susceptible individuals before symptoms occur (e.g., based on a history of symptoms and / or based on genetic or other susceptibility factors). Treatment may also continue after symptoms have subsided, for example, to prevent or delay their recurrence.

[0320] The provided compounds are inhibitors of Wee1A kinase and are therefore used to treat one or more conditions associated with Wee1A kinase activity. Accordingly, in some embodiments, this disclosure provides a method for treating Wee1A kinase-mediated conditions, comprising the step of administering the disclosed compounds or pharmaceutically acceptable compositions thereof to a patient in need.

[0321] Some of the provided compounds also exhibit potent inhibitory activity against Myt1 kinase, and are therefore dual inhibitors that can be used to treat one or more conditions associated with the activity of both Wee1A kinase and Myt1 kinase. Therefore, in some embodiments, this disclosure provides a method for treating Wee1A kinase / Myt1 kinase-mediated conditions, comprising the step of administering a compound of this disclosure as a dual inhibitor or a pharmaceutically acceptable composition thereof to a patient in need.

[0322] As used herein, the term "Wee1A kinase-mediated" condition or disorder means any disease or other harmful disorder in which Wee1A kinase or its mutants are known to function. Therefore, another embodiment of this disclosure relates to treating or reducing the severity of one or more diseases in which Wee1A kinase or its mutants are known to function. Specifically, this disclosure relates to methods of treating or reducing the severity of diseases or disorders selected from proliferative disorders, said methods comprising administering a compound or composition according to this disclosure to a patient in need.

[0323] As used herein, the term "Wee1A kinase / Myt1 kinase-mediated" condition or disorder means any disease or other harmful disorder in which mutants of Wee1A kinase and Myt1 kinase, or any one or both of the foregoing, are known to function. Therefore, another embodiment of this disclosure relates to treating or reducing the severity of one or more diseases in which mutants of Wee1A kinase and Myt1 kinase, or any one or both of the foregoing, are known to function. Specifically, this disclosure relates to methods of treating or reducing the severity of diseases or disorders selected from proliferative disorders, said methods comprising administering a compound or composition according to this disclosure to a patient in need.

[0324] In some embodiments, this disclosure provides a method for inhibiting Wee1A kinase activity in a subject, comprising administering to the subject an effective amount of a compound of this disclosure or a pharmaceutically acceptable composition.

[0325] In some embodiments, this disclosure provides a method for inhibiting the activity of Wee1A kinase and Myt1 kinase in a subject, comprising administering to the subject an effective amount of a compound of this disclosure or a pharmaceutically acceptable composition as a dual inhibitor.

[0326] In some embodiments, this disclosure provides a method for treating or reducing the severity of one or more conditions selected from cancer, comprising the step of administering to a subject an effective amount of a compound of this disclosure or a pharmaceutically acceptable composition thereof. In some embodiments, the cancer is associated with a solid tumor.

[0327] In some embodiments, this disclosure provides a method of treating a subject suffering from cancer or other cell growth disorders characterized by aberrant Wee1A kinase activity, comprising administering to the subject an effective amount of a compound of this disclosure or a pharmaceutically acceptable composition thereof. In some embodiments, aberrant Wee1A kinase activity includes elevated or overexpressed activity compared to a disease-free state, or unwanted activity. In some such embodiments, aberrant Wee1A kinase activity may include disrupted p53 activity, Cdk1 activity, Cdk2 activity, replication stress, mitotic alterations, and DNA damage. In some embodiments, the subject suffers from cancer associated with p53 inactivation.

[0328] In some embodiments, this disclosure provides a method of treating a subject suffering from cancer or other cell growth disorders characterized by both abnormal Wee1A kinase activity and abnormal Myt1 kinase activity, comprising administering to the subject an effective amount of a compound of this disclosure or a pharmaceutically acceptable composition thereof as a dual inhibitor. In some embodiments, abnormal Wee1A kinase activity and abnormal Myt1 kinase activity include elevated or overexpressed activity compared to a disease-free state, or undesirable activity. In some such embodiments, abnormal Wee1A kinase activity and abnormal Myt1 kinase activity may include disturbed Cdk1 activity, replication stress, mitotic alterations, and DNA damage. In some embodiments, the subject to be treated has previously been treated with a monospecific Wee1A kinase inhibitor or a Myt1 kinase inhibitor (neither of which is a dual inhibitor) and has developed resistance to such treatment or is refractory to such treatment. For example, such subjects may be resistant to or refractory to Myt1 kinase inhibitor RP-6306 or Wee1A kinase inhibitors AZD1775, Debio0123 or ZnC3.

[0329] In some embodiments, the cancer to be treated by the compounds disclosed herein is selected from brain cancer, cervical brain cancer, heart cancer, gastrointestinal cancer, esophageal cancer, thyroid cancer, small cell carcinoma, non-small cell carcinoma, breast cancer, lung cancer, gastric cancer, gallbladder / choleduct cancer, liver cancer, pancreatic cancer, colon cancer, rectal cancer, ovarian cancer, choriocarcinoma, endometrial cancer, cervical cancer, renal pelvis / ureter cancer, bladder cancer, prostate cancer, penile cancer, testicular cancer, fetal cancer, Wilms' disease, skin cancer, malignant melanoma, neuroblastoma, osteosarcoma, Ewing's tumor, soft tissue sarcoma, acute leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, polycythemia vera, malignant lymphoma, multiple myeloma, Hodgkin's lymphoma, and non-Hodgkin's lymphoma. In some embodiments, the subject has a cancer selected from serous uterine carcinoma and renal cancer.

[0330] In some implementations, breast cancer is selected from ductal carcinoma in situ (DCIS), invasive ductal carcinoma (IDC), lobular carcinoma in situ (LCIS), invasive lobular carcinoma (ILC), triple-negative breast cancer (TNBC), inflammatory breast cancer (IBC), metastatic breast cancer (MBC), medullary carcinoma, tubular carcinoma, mucinous carcinoma (colloid carcinoma), and nipple or breast Paget's disease (commonly referred to as Paget's disease).

[0331] 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.

[0332] In some implementations, ovarian cancer is selected from epithelial ovarian cancer, germ cell tumors, and stromal cell tumors.

[0333] In some implementations, the gastric cancer is selected from adenocarcinoma, lymphoma, gastrointestinal stromal tumor (GIST), carcinoid tumor, and hereditary (familial) diffuse gastric cancer.

[0334] In some embodiments, the esophageal cancer is selected from squamous cell carcinoma, small cell carcinoma, and adenocarcinoma. In some embodiments, the esophageal cancer is squamous cell carcinoma. In some embodiments, the esophageal cancer is adenocarcinoma.

[0335] In some implementations, the lung cancer is selected from non-small cell lung cancer, pulmonary nodules, small cell lung cancer, and mesothelioma. In some implementations, the lung cancer is non-small cell lung cancer.

[0336] In some implementations, colorectal cancer is selected from adenocarcinoma, gastrointestinal stromal tumor (GIST), lymphoma, carcinoid, Turcot syndrome, Peutz-Jeghers syndrome (PJS), familial colorectal cancer (FCC), and juvenile colonic adenomatous polyposis.

[0337] In some implementations, cancer is associated with dysregulation of cyclin E1. In some implementations, ovarian cancer is associated with dysregulation of cyclin E1.

[0338] In some implementations, cancer is associated with p53 dysregulation. In some implementations, cancers associated with p53 dysregulation are selected from brain cancer, cervical brain cancer, heart cancer, gastrointestinal cancer, esophageal cancer, thyroid cancer, small cell carcinoma, non-small cell carcinoma, breast cancer, lung cancer, stomach cancer, gallbladder / choleduct cancer, liver cancer, pancreatic cancer, colon cancer, rectal cancer, ovarian cancer, choriocarcinoma, endometrial cancer, cervical cancer, renal pelvis / ureter cancer, bladder cancer, prostate cancer, penile cancer, testicular cancer, fetal cancer, Wilms' disease, skin cancer, malignant melanoma, neuroblastoma, osteosarcoma, Ewing's tumor, soft tissue sarcoma, acute leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, polycythemia vera, malignant lymphoma, multiple myeloma, Hodgkin's lymphoma, and non-Hodgkin's lymphoma. In some such implementations, cancers associated with p53 dysregulation are selected from serous uterine carcinoma and kidney cancer.

[0339] In some implementations, cancer is associated with Cdk1 dysregulation. In some implementations, cancers associated with Cdk1 dysregulation are selected from brain cancer, cervical brain cancer, heart cancer, gastrointestinal cancer, esophageal cancer, thyroid cancer, small cell carcinoma, non-small cell carcinoma, breast cancer, lung cancer, stomach cancer, gallbladder / choleduct cancer, liver cancer, pancreatic cancer, colon cancer, rectal cancer, ovarian cancer, choriocarcinoma, endometrial cancer, cervical cancer, renal pelvis / ureter cancer, bladder cancer, prostate cancer, penile cancer, testicular cancer, fetal cancer, Wilms' disease, skin cancer, malignant melanoma, neuroblastoma, osteosarcoma, Ewing's tumor, soft tissue sarcoma, acute leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, polycythemia vera, malignant lymphoma, multiple myeloma, Hodgkin's lymphoma, and non-Hodgkin's lymphoma. In some such implementations, cancers associated with Cdk1 dysregulation are selected from serous uterine carcinoma and kidney cancer.

[0340] In some implementations, cancer is associated with Cdk2 dysregulation. In some implementations, cancers associated with Cdk2 dysregulation are selected from brain cancer, cervical brain cancer, heart cancer, gastrointestinal cancer, esophageal cancer, thyroid cancer, small cell carcinoma, non-small cell carcinoma, breast cancer, lung cancer, stomach cancer, gallbladder / choleduct cancer, liver cancer, pancreatic cancer, colon cancer, rectal cancer, ovarian cancer, choriocarcinoma, endometrial cancer, cervical cancer, renal pelvis / ureter cancer, bladder cancer, prostate cancer, penile cancer, testicular cancer, fetal cancer, Wilms' disease, skin cancer, malignant melanoma, neuroblastoma, osteosarcoma, Ewing's tumor, soft tissue sarcoma, acute leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, polycythemia vera, malignant lymphoma, multiple myeloma, Hodgkin's lymphoma, and non-Hodgkin's lymphoma. In some such implementations, cancers associated with Cdk1 dysregulation are selected from serous uterine carcinoma and kidney cancer.

[0341] Depending on the specific ailment or disease to be treated, additional therapeutic agents that are typically administered to treat that ailment may also be present in the compositions disclosed herein. As used herein, additional therapeutic agents that are typically administered to treat a specific ailment or disease are referred to as “suitable for the ailment or disease to be treated”.

[0342] For example, the compounds of this disclosure or pharmaceutically acceptable combinations thereof may be administered in combination with chemotherapeutic agents to treat proliferative diseases and cancers. Examples of known chemotherapeutic agents include, but are not limited to, doxorubicin, dexamethasone, vincristine, cyclophosphamide, fluorouracil, topotecan, taxol, interferon, platinum derivatives, taxanes (e.g., paclitaxel), vinca alkaloids (e.g., vincristine), anthracyclines (e.g., doxorubicin), epipodophyllotoxins (e.g., etoposide), cisplatin, mTOR inhibitors (e.g., rapamycin), methotrexate, actinomycin D, dolalastatin 10, colchicine, emetine, trimetrexate, metoprine, cyclosporine, daunorubicin, teniposide, amphotericin B, alkylating agents (e.g., chlorambucil), 5-fluorouracil, camptothecin, cisplatin, metronidazole, and Gleevec™, etc. In other embodiments, the compounds of this disclosure are administered in combination with biological agents such as Avastin or VECTIBIX.

[0343] In some embodiments, the compounds of this disclosure or pharmaceutically acceptable combinations thereof are administered in combination with an agent selected from the group consisting of: fasudil, sirolimus, imatinib, gefitinib, erlotinib, sorafenib, sunitinib, dasatinib, lapatinib, nilotinib, temsirolimus, everolimus, pazopanib, ruxolitinib, and vandetani. b) Vemurafenib, Crizotinib, Icotinib, Axitinib, Tofacitinib, Bosutinib, Cabozantinib, Ponatinib, Regorafenib, Afatinib, Dabrafenib, Trametinib, Ibrutinib, Nintedanib, Idelalisib, Ceritinib, Apatinibrivoceranib, ripasudil, alectinib, cobimetinib, lenvatinib, palbociclib, radotinib, osimertinib, olmutinib, neratinib, ribociclib, copanlisib, abemacic lib), acalabrutinib, midostaurin, brigatinib, baricitinib, netarsudil, tivozanib, simotinib, fostamatinib, encorafenib, binimetinib, carticinib, duvelixib Elisib, Dacomitinib, Lolatinib, Larotrectinib, Gilteritinib, Pyrotinib, Fruquintinib, Erdafitinib, Alpelisib, Umbralisib, Leniolisib, Pexidartinib, Entricinib (entrectinib), upadacitinib, fedratinib, zanubrutinib, flumatinib, peficitinib, delgocitinib, avapritinib, selumetinib, tucatinib, pemigatinib, capmatinibThe following are listed: tabrecta, serpercatinib, ripretinib, tirabrutinib, almonertinib, pralsetinib, filgotinib, tirbanibulin, orelabrutinib, tepotinib, and trilaciclib. See also www.ppu.mrc.ac.uk / list-clinically-approved-kinase-inhibitors List of clinically approved kinase inhibitors | MRC Protein Phosphorylation Ubiquitylation Unit The entire text is incorporated into this paper by reference.

[0344] In some embodiments, the compounds of this disclosure or pharmaceutically acceptable combinations thereof are administered in combination with an antiproliferative agent or chemotherapeutic agent selected from any one or more of the following: abarelix, interleukin, alemtuzumab, alitretinoin, allopurinol, hexamethylmelamine, aifostine, anastrozole, arsenic trioxide, asparaginase, azacitidine, and BCG. Liver, bevacizumab, fluorouracil, bexarotene, bleomycin, bortezomib, busulfan, calusterone, capecitabine, camptothecin, carboplatin, carmustine, celecoxib, cetuximab, chlorambucil, cladribine, clofarabine, cyclophosphamide, cytarabine, actinomycin D, darbepoetin alfa, daunorubicin, decitabine, denileukin, dexrazoxane, docetaxel, doxorubicin (neutral), doxorubicin hydrochloride, dromostanolone propionate propionate, epirubicin, epoetin alpha, erlotinib, estramustine, etoposide phosphate, etoposide, exemestane, filgrastim, fludarabine, fulvestrant, gefitinib, gemcitabine, gemtuzumab, goserelin acetate, histrelin acetateAcetate), hydroxyurea, ibritumomab, idarubicin, ifosfamide, imatinib mesylate, interferon alpha-2a, interferon alpha-2b, irinotecan, lenalidomide, letrozole, leucovorin, leuprolide acetate, levamisole, lomustine, megestrol acetate 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 maleatemaleate), 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.

[0345] In some embodiments, the compounds of this disclosure or pharmaceutically acceptable compositions thereof are co-administered with a pharmaceutically acceptable Myt1 kinase inhibitor. In some such embodiments, the Myt1 kinase inhibitor is RP-6306.

[0346] In some embodiments, the compounds of this disclosure or pharmaceutically acceptable combinations thereof are co-administered with a pharmaceutically acceptable DNA-damaging agent.

[0347] In some embodiments, the compounds of this disclosure or pharmaceutically acceptable combinations thereof are co-administered with radiation.

[0348] In some embodiments, the compounds of this disclosure or pharmaceutically acceptable combinations thereof are administered in combination with monoclonal antibodies or siRNA therapeutic agents.

[0349] In some embodiments, the compounds of this disclosure or pharmaceutically acceptable compositions thereof are administered in combination with a targeted therapy selected from (i) inhibitors of kinases selected from MET, MEK, mTOR, FLT3, BRAF, KIT, PDGFR, FDFR, PI3K, EGFR, AKT, and KRAS; (ii) inhibitors of fusion kinases such as BCR-ABL, ALK, RET and ROS, JAK, CDK4 / 6, and KRAS; (iii) epigenetic regulators such as HDAC inhibitors; (iv) immuno-oncology agents such as those targeting PD1, PDL1, and CTLA4; and (v) antibody-drug conjugates such as those targeting Her2, CD4, and PD1. 38. Those of BCMA, CD19, connexin 4, trop2, CD79, and CD22; (vi) bispecific T cell binders (BiTE); (vii) transcription factor regulators, such as those targeting IKZF (i.e., IMiD and EZH2); (viii) steroid receptor regulators, such as those targeting AR and ER; and (ix) proteasome inhibitors, such as bortezomib, ixazomib, and carfilzomib; and (x) apoptosis-targeting agents, such as inhibitors of BCL-2, BCL-XL, MCL1, IAP, or TRAIL / death receptor agonists.

[0350] In some embodiments, the compounds of this disclosure or pharmaceutically acceptable combinations thereof are administered in combination with an inhibitor of a DNA repair protein, rather than Wee1A kinase or Myt1 kinase. Such inhibitors include those that inhibit one or more of the following: CHK1, CHK2, ATM, ATR, Pol θ, CDC7, DNAPK, PLK1, WRN, PARP, and Aurora A / B.

[0351] These additional agents may be administered separately from the composition containing the compounds of the present invention as part of a multi-dosing regimen. Alternatively, those agents may be part of a single dosage form, mixed with the compounds of the present disclosure in a single composition. If administered as part of a multi-dosing regimen, the two active agents may be delivered simultaneously, sequentially, or at intervals between each other, for example, at intervals of one, two, three, four, five, six, seven, eight, nine, ten, eleven, or twelve hours.

[0352] In some such embodiments, the compounds of this disclosure or pharmaceutically acceptable salts thereof, particularly those compounds that do not have significant Myt1 kinase activity (e.g., compounds not rated A or B in the Myt1 kinase binding assay disclosed in Example 6 herein), are administered as part of a multiple-dose regimen together with a pharmaceutically acceptable Myt1 kinase inhibitor. In some embodiments, the compounds of this disclosure or pharmaceutically acceptable compositions thereof are administered as part of a multiple-dose regimen together with a Myt1 kinase inhibitor selected from RP-6306.

[0353] In some embodiments, the compounds of this disclosure or pharmaceutically acceptable salts thereof, particularly those compounds that do not have significant Myt1 kinase activity, are administered to a subject, wherein the Myt1 kinase inhibitor is used as first-line or second-line therapy. In some embodiments, the compounds of this disclosure or pharmaceutically acceptable salts thereof are administered to a subject, wherein the Myt1 kinase inhibitor is used as first-line therapy. In some embodiments, the compounds of this disclosure or pharmaceutically acceptable salts thereof are administered to a subject, wherein a Myt1 kinase inhibitor selected from RP-6306 is used as first-line therapy.

[0354] In some embodiments, the compounds of this disclosure or pharmaceutically acceptable salts thereof, particularly those compounds that do not have significant Myt1 kinase activity, are administered to a subject, wherein the Myt1 kinase inhibitor is used as a second-line therapy. In some embodiments, the compounds of this disclosure or pharmaceutically acceptable salts thereof are administered to a subject, wherein a Myt1 kinase inhibitor selected from RP-6306 is used as a second-line therapy.

[0355] As used herein, the terms “combination,” “combined,” “co-administered,” and related terms refer to the simultaneous or sequential administration of a therapeutic agent according to this disclosure. For example, the compound of this disclosure may be administered simultaneously or sequentially with another therapeutic agent in separate unit dosage forms or together in a single unit dosage form. Thus, this disclosure provides a single unit dosage form comprising the provided compound, an additional therapeutic agent, and a pharmaceutically acceptable carrier, adjuvant, or mediator.

[0356] The amounts of both the compounds of the present invention, which can be combined with a carrier substance to produce a single dosage form, and additional therapeutic agents (in those compositions containing additional therapeutic agents as described above) will vary depending on the host being treated and the specific administration method. Preferably, the compositions of the present disclosure are formulated such that the dosage of the present invention, ranging from 0.001 to 100 mg / kg body weight / day, can be administered.

[0357] In those compositions that include an additional therapeutic agent, the additional therapeutic agent and the compounds disclosed herein can work synergistically. Therefore, the amount of the additional therapeutic agent in such compositions will be less than that required for a single therapy using only the therapeutic agent. In such compositions, a dose of the additional therapeutic agent of 0.001-1,000 µg / kg body weight / day can be administered.

[0358] The amount of additional therapeutic agent present in the compositions disclosed herein will not exceed the amount normally applied in compositions containing the therapeutic agent as the sole active agent. Preferably, the amount of additional therapeutic agent in the compositions disclosed herein ranges from about 50% to 100% of the amount normally present in compositions containing the agent as the sole active agent.

[0359] In some embodiments, this disclosure provides a method for inhibiting Wee1A kinase in vitro. In some such embodiments, the amount of Wee1A kinase inhibition is assessed based on a competitive ATP binding assay.

[0360] In some embodiments, this disclosure provides a method for inhibiting Wee1A kinase in biological samples.

[0361] In some embodiments, this disclosure provides methods for inhibiting both Wee1A kinase and Myt1 kinase in biological samples.

[0362] In some embodiments, this disclosure provides methods for inhibiting both Wee1A kinase and Myt1 kinase in vitro. In some such embodiments, the amount of Wee1A kinase and Myt1 kinase inhibition is assessed based on a competitive ATP binding assay.

[0363] In some embodiments, this disclosure provides a method for evaluating Cdk1 phosphorylation in cells, which includes contacting the cells with the compounds described herein. In one embodiment, the contacting step includes incubating the cells with the compounds provided herein. In some such embodiments, the cells are incubated for at least 4 hours. In some embodiments, the cells may include DAOY medulloblastoma cells.

[0364] Example

[0365] General methods

[0366] Unless otherwise specified, reagents and solvents were purchased from commercial suppliers and used as is. Solvents were dried over a molecular sieve at 4 Å. The reaction was carried out in a glass vial or round-bottom flask with a magnetic stir bar and heated with a stirring plate. Solvents were removed by rotary evaporation, vacuum centrifugation, or lyophilization. The reaction progress was monitored by LC-MS or thin-layer chromatography (TLC). Aluminum-lined TLC plates (60°F) were illuminated by UV light (254 nm). 254(Visible)

[0367] At 25°C, at 500 MHz (500 MHz) with a 5 mm iProbe BBF / H / D probe. 1 H NMR and at 126 MHz 13 (C NMR) Bruker Avance Neo spectrometer or equipped with a 5 mm 1H / 13C auto-switching gradient probe at 400 MHz (400 MHz) 1 H NMR and at 101 MHz 13 Recorded on a Varian Inova spectrometer (C NMR) 1 H NMR and 13 C10 NMR spectroscopy. Chloroform- d (δ H 7.27 ppm), dimethyl sulfoxide - d 6 (δ H 2.50 ppm), acetonitrile- d 3 (δ H 1.95 ppm) or methanol- d 4 (δ H The central peak (3.31 ppm) was used as an internal reference. The spectrum was processed using commercial software.

[0368] Using a reversed-phase C18 column, elution was performed with acetonitrile and water (containing 0.1% TFA or 0.03% ammonia). The mass spectrometer was operated in ES (+ or -) ionization mode, and LC-MS was obtained on the instrument.

[0369] Rapid chromatography was performed on silica gel or C18 functionalized silica using an automated rapid purification system equipped with a diode array detector (200-400 nm), eluting with a gradient of ethyl acetate and petroleum ether or methanol and dichloromethane (containing 0.03% ammonia).

[0370] Purity analysis was performed using a reverse-phase C18 HPLC column, eluted with acetonitrile and water (containing 0.1% TFA or 0.03% ammonia). UV traces were recorded at 220 nm. Preparative HPLC purification was also performed using a reverse-phase C18 column, eluted with acetonitrile and water (containing 0.1% TFA or 0.03% ammonia).

[0371] Specify the stereochemical configuration of any enantiomer or diastereomer separated by chiral chromatography. Absolute configuration unknown.

[0372] abbreviation:

[0373]

[0374]

[0375] Example 1. Synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(piperidin-4-ylamino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid (TFA salt of compound 124) and 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid (TFA salt of compound 138)

[0376]

[0377] 4-((6-bromopyridin-2-yl)amino)piperidine-1-carboxylic acid tert-butyl ester

[0378] At room temperature, N,N Diisopropylethylamine (3 equivalents) and tert-butyl 4-aminopiperidine-1-carboxylate (1 equivalent) were added to a solution of 2,6-dibromopyridine (1 equivalent) in DMSO (0.8 mol / L). The reaction mixture was heated to 90 °C and stirred for several days until LCMS indicated complete conversion. The reaction mixture was diluted with brine and a saturated aqueous solution of NaHCO3 and extracted with ethyl acetate (×3). The combined organic layers were dried using a phase separator and concentrated under reduced pressure to give an oil. This material was used without further purification or purified by rapid chromatography.

[0379] Alternatively, cesium carbonate (3 equivalents) and tert-butyl 4-aminopiperidine-1-carboxylate (1 equivalent) were added to a solution of 2,6-dibromopyridine (1 equivalent) in anhydrous dimethylformamide (0.4 mol / L) and stirred at 100 °C until LCMS indicated complete conversion, usually overnight. The reaction mixture was diluted with brine and a saturated aqueous solution of NaHCO3 and extracted with ethyl acetate (×3). The combined organic layers were dried using a phase separator and concentrated under reduced pressure to give an oil. This material was used without further purification or purified by rapid chromatography.

[0380] 4-((6-(2-allyl-6-(methylthio)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl) tert-butyl pyridin-2-yl)amino)piperidine-1-carboxylate

[0381] At room temperature, CuI (1.2 equivalents) and subsequently N,N'Dimethylethylenediamine (1 equivalent) was added to a stirred, degassed suspension of 4-((6-bromopyridin-2-yl)amino)piperidin-1-carboxylic acid tert-butyl ester (1 equivalent), 6-(methylthioalkyl)-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (1 equivalent) and cesium carbonate (3 equivalents) in dioxane (0.3 mol / L). The reaction was heated to 90°C overnight in a sealed vial. The reaction mixture was diluted with water and a few drops of ammonia solution (28%) and then extracted with ethyl acetate (×3). The combined organic layers were dried using a phase separator and concentrated under reduced pressure to give an oil. This material was used without further purification or purified by rapid chromatography.

[0382] 2-Allyl-6-((4-chlorophenyl)amino)-1-(6-(piperidin-4-ylamino)pyridin-2-yl)-1,2-dihydro- 3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 124)

[0383] At room temperature, mCPBA (approximately 75%, 1.2 equivalents) was added to a stirred solution of 4-((6-(2-allyl-6-(methylthio)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyridin-2-yl)amino)piperidine-1-carboxylic acid tert-butyl ester (1 equivalent) in dichloromethane (0.4 mol / L). The mixture was stirred until LCMS indicated complete conversion to the corresponding sulfoxides (large amount) and sulfones (small amount), typically within 1 hour.

[0384] 1-Amino-4-chlorobenzene (1 equivalent) is added to the reaction mixture and the resulting mixture is heated to 40 °C until LCMS indicates complete conversion, usually overnight. Alternatively, dichloromethane is removed under reduced pressure and the residue is redissolved in anhydrous acetonitrile (0.6 mol / L). 1-Amino-4-chlorobenzene (1 equivalent) is then added, and the resulting mixture is heated to 60 °C until LCMS indicates complete conversion, usually overnight. The reaction mixture is concentrated, the residue is diluted with ethyl acetate, an aqueous solution of NaOH (1 M) is added, and the product is extracted with ethyl acetate (×3). The combined organic layers are dried using a phase separator and concentrated to the residue under reduced pressure.

[0385] At room temperature, trifluoroacetic acid (10-20% by volume) was added to a stirred solution of the above material (1 equivalent) in anhydrous dichloromethane (0.4 mol / L). The resulting solution was stirred until LCMS indicated complete conversion, typically within 1 hour. The reaction mixture was concentrated and purified by reversed-phase chromatography. The purified fractions were collected and lyophilized to give compound 124. Yield: 29 mg, 49%, as a yellow solid. HPLC purity (220 nm): 100%.

[0386] 2-Allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1, 2-Dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 138)

[0387] Formaldehyde (38% aqueous solution, 2 equivalents) was added to a stirred solution of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(piperidin-4-ylamino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (1 equivalent) in THF (0.6 mol / L) at room temperature. If the starting material was in the form of trifluoroacetate, diisopropylethylamine (1 equivalent) was added. The reaction mixture was stirred at room temperature for 30 minutes, and then sodium triacetoxyborohydride (3 equivalents) was added in portions. The mixture was stirred until LCMS indicated complete conversion, which in some cases required the addition of formaldehyde and sodium triacetoxyborohydride. The reaction mixture was concentrated and purified by reversed-phase chromatography. The purified fractions were collected and lyophilized to give compound 128. Yield: 7.4 mg, 63%, as a yellow solid. HPLC purity (220 nm): 98%.

[0388] Example 2. Synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(piperidin-4-yloxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid (TFA salt of compound 141) and 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)oxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid (TFA salt of compound 142)

[0389]

[0390] 4-((6-bromopyridin-2-yl)oxy)piperidine-1-carboxylic acid tert-butyl ester

[0391] Sodium hydride was added to a stirred solution of tert-butyl 4-hydroxypiperidine-1-carboxylate (1 equivalent) in tetrahydrofuran (0.5 mol / mL) at 0 °C. A solution of 2,6-dibromopyridine (1 equivalent) in THF (0.5 mol / mL) was added to the mixture. After complete addition, the mixture was brought to room temperature and stirred until LCMS indicated complete conversion, usually overnight. The reaction mixture was concentrated, diluted with brine and a saturated aqueous solution of NaHCO3, and extracted with ethyl acetate (×3). The organic layer was washed with brine and then filtered through a phase separator. The organic layer was concentrated to the residue under reduced pressure. The residue was purified by rapid chromatography.

[0392] 4-((6-(2-allyl-6-(methylthio)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl) tert-butyl pyridin-2-yl)oxy)piperidine-1-carboxylate

[0393] At room temperature, CuI (1.2 equivalents) and subsequently N,N'Dimethylethylenediamine (1 equivalent) was added to a stirred, degassed suspension of 4-((6-bromopyridin-2-yl)oxy)piperidin-1-carboxylic acid tert-butyl ester (1 equivalent), 6-(methylthioalkyl)-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (1 equivalent) and cesium carbonate (3 equivalents) in dioxane (0.3 mol / L). The reaction was heated to 90°C overnight in a sealed vial. The reaction mixture was diluted with water and a few drops of ammonia solution (28%) and then extracted with ethyl acetate (×3). The combined organic layers were dried using a phase separator and concentrated under reduced pressure to give an oil. This material was used without further purification or purified by rapid chromatography.

[0394] 2-Allyl-6-((4-chlorophenyl)amino)-1-(6-(piperidin-4-yloxy)pyridin-2-yl)-1,2-dihydro- 3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 141)

[0395] At room temperature, mCPBA (approximately 75%, 1.2 equivalents) was added to a stirred solution of tert-butyl 4-((6-(2-allyl-6-(methylthio)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyridin-2-yl)oxy)piperidine-1-carboxylate (1 equivalent) in dichloromethane (0.4 mol / L). The mixture was stirred until LCMS indicated complete conversion to the corresponding sulfoxides (large amount) and sulfones (small amount), typically within 1 hour.

[0396] 1-Amino-4-chlorobenzene (1 equivalent) is added to the reaction mixture and the resulting mixture is heated to 40 °C until LCMS indicates complete conversion, usually overnight. Alternatively, dichloromethane is removed under reduced pressure and the residue is redissolved in anhydrous acetonitrile (0.6 mol / L). 1-Amino-4-chlorobenzene (1 equivalent) is then added, and the resulting mixture is heated to 60 °C until LCMS indicates complete conversion, usually overnight. The reaction mixture is concentrated, the residue is diluted with ethyl acetate, an aqueous solution of NaOH (1 M) is added, and the product is extracted with ethyl acetate (×3). The combined organic layers are dried using a phase separator and concentrated to the residue under reduced pressure.

[0397] At room temperature, trifluoroacetic acid (10-20% by volume) was added to a stirred solution of the above material (1 equivalent) in anhydrous dichloromethane (0.4 mol / L). The resulting solution was stirred until LCMS indicated complete conversion, typically within 1 hour. The reaction mixture was concentrated and purified by reversed-phase chromatography. The purified fractions were collected and lyophilized to give compound 141. Yield: 15 mg, 31%, as a solid. HPLC purity (220 nm): 98%.

[0398] 2-Allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)oxy)pyridin-2-yl)-1, 2-Dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 142)

[0399] Formaldehyde (38% aqueous solution, 2 equivalents) was added to a stirred solution of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(piperidin-4-yloxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (1 equivalent) in THF (0.6 mol / L) at room temperature. If the starting material was in the form of trifluoroacetate, diisopropylethylamine (1 equivalent) was added. The reaction mixture was stirred at room temperature for 30 minutes, and then sodium triacetoxyborohydride (3 equivalents) was added in portions. The mixture was stirred until LCMS indicated complete conversion, which in some cases required the addition of formaldehyde and sodium triacetoxyborohydride. The reaction mixture was concentrated and purified by reversed-phase chromatography. The purified fractions were collected and lyophilized to give compound 142. Yield: 42 mg, 63%, as solid. HPLC purity (220 nm): 97%.

[0400] Example 3. Synthesis of 1-{6-[(3R)-1-azabicyclo[2.2.2]octyl-3-yloxy]pyridin-2-yl}-6-[(1-methyl-1H-pyrazol-4-yl)amino]-2-(propen-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid (TFA salt of compound 172)

[0401]

[0402] (R)-3-((6-bromopyridin-2-yl)oxy)quinine ring

[0403] At room temperature, N,N Diisopropylethylamine (3 equivalents) and (R)-quininecyclo-3-ol (1 equivalent) were added to a solution of 2,6-dibromopyridine (1 equivalent) in DMSO (0.8 mol / L). The reaction mixture was heated to 90 °C and stirred for several days until LCMS indicated complete conversion. The reaction mixture was diluted with brine and a saturated aqueous solution of NaHCO3 and extracted with ethyl acetate (×3). The combined organic layers were dried using a phase separator and concentrated under reduced pressure to give an oil. This material was used without further purification or purified by rapid chromatography.

[0404] Alternatively, cesium carbonate (3 equivalents) and (R)-quininecyclo-3-ol (1 equivalent) were added to a solution of 2,6-dibromopyridine (1 equivalent) in anhydrous dimethylformamide (0.4 mol / L) and stirred at 100 °C until LCMS indicated complete conversion, usually overnight. The reaction mixture was diluted with brine and a saturated aqueous solution of NaHCO3 and extracted with ethyl acetate (×3). The combined organic layers were dried using a phase separator and concentrated under reduced pressure to give an oil. This material was used without further purification or purified by rapid chromatography.

[0405] (R)-2-Allyl-6-(methylthio)-1-(6-(quininyl-3-yloxy)pyridin-2-yl)-1,2-dihydro-3H- Pyrazolo[3,4-d]pyrimidin-3-one

[0406] At room temperature, CuI (1.2 equivalents) and subsequently N,N' Dimethylethylenediamine (1 equivalent) was added to a stirred, degassed suspension of (R)-3-((6-bromopyridin-2-yl)oxy)quinine ring (1 equivalent), 6-(methylthioalkyl)-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (1 equivalent) and cesium carbonate (3 equivalents) in dioxane (0.3 mol / L). The reaction was heated to 90°C overnight in a sealed vial. The reaction mixture was diluted with water and a few drops of ammonia solution (28%) and then extracted with ethyl acetate (×3). The combined organic layers were dried using a phase separator and concentrated under reduced pressure to give an oil. This material was used without further purification or purified by rapid chromatography.

[0407] (R)-2-Allyl-6-((1-Methyl-1H-pyrazol-3-yl)amino)-1-(6-(quininecyclo-3-yloxy)pyridine) (Pinidin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 172)

[0408] At room temperature, mCPBA (approximately 75%, 1.2 equivalents) was added to a stirred solution of methanesulfonic acid (2 equivalents) and (R)-2-allyl-6-(methylthio)-1-(6-(quininecyclo-3-yloxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (1 equivalent) in dichloromethane (0.6 mol / L). The mixture was stirred until LCMS indicated complete conversion to the corresponding sulfoxides (large amount) and sulfones (small amount), typically within 1 hour.

[0409] 1-Methyl-1H-pyrazole-4-amine (1 equivalent) was added to the reaction mixture, and the resulting mixture was heated to 40 °C until LCMS indicated complete conversion, usually overnight. Alternatively, dichloromethane was removed under reduced pressure, and the residue was redissolved in anhydrous acetonitrile (0.6 mol / L). Then, 1-methyl-1H-pyrazole-4-amine (1 equivalent) was added, and the resulting mixture was heated to 60 °C until LCMS indicated complete conversion, usually overnight. The mixture was concentrated to dryness under reduced pressure and purified by reversed-phase chromatography. The pure fractions were collected and lyophilized to give the product. Yield: 53 mg, 38%, as a yellow solid. HPLC purity (220 nm): 98%.

[0410] Other compounds disclosed herein can be synthesized or synthesized using one or more of the following synthetic routes described in Examples 1-3: different reactive aromatic rings for initiating 2,6-dibromopyridine, different amines in step 1A, different alcohols in step 1B, or different aromatic cyclic amines in steps 3A or 3B. Those skilled in the art of medicinal chemistry will be able to synthesize the disclosed compounds without extensive experimentation by adapting the disclosed examples.

[0411] Example 4. Synthesis of 2-allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 370)

[0412] 4-{6-[6-(4-bromophenylamino)-3-oxo-2-(prop-2-enyl)-1,2-dihydro-3H-1,2,5,7-tetraazine} [Indene-1-yl]pyridin-2-yloxy}piperidine-1-carboxylic acid tert-butyl ester

[0413] Under nitrogen atmosphere, at room temperature, 83.1 mg (assumed 0.481 mmol) of mCPBA (<77% pure) in DCM (0.5 mL) was added to a stirred solution of 4-{6-[2-allyl-6-(methylthio)-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazainden-1-yl]-2-pyridyloxy}-1-piperidinecarboxylate tert-butyl ester (200 mg, 0.401 mmol) in DCM (5 mL). The reaction was controlled by LC-MS. After 15 minutes, DCM was removed under vacuum, and the crude product was dissolved in MeCN (5 mL), followed by the addition of p-bromoaniline (69 mg, 0.401 mmol). The reaction mixture was stirred in a sealed vial at 60 °C. After 96 hours, the reaction mixture was cooled to room temperature, and mCPBA quenched with 1 M NaOH (5 mL) was added dropwise. The aqueous phase was extracted with EtOAc (3 x 20 mL) and washed with brine (20 mL). The combined organic layers were dried (MgSO4) and concentrated under reduced pressure to give the product as a yellow powder. The crude material was dissolved in DCM (5 mL), loaded onto a 10 g silica gel column, and purified by rapid chromatography (0-100%, EtOAc: PE) to give 4-{6-[6-(4-bromophenylamino)-3-oxo-2-(prop-2-enyl)-1,2-dihydro-3- H -1,2,5,7-Tetraazaindene-1-yl]pyridin-2-yloxy}piperidine-1-carboxylic acid Uncle Butyl ester [180 mg, 59%] is a light yellow solid.

[0414] 6-(4-Bromophenylamino)-1-[6-(piperidin-4-yloxy)pyridin-2-yl]-2-(prop-2-enyl)-1,2-di Hydrogen-3H-1,2,5,7-tetraazainden-3-one

[0415] 4-{6-[6-(4-bromophenylamino)-3-oxo-2-(prop-2-enyl)-1,2-dihydro-3- H -1,2,5,7-Tetraazaindene-1-yl]pyridin-2-yloxy}piperidine-1-carboxylic acid Uncle A solution of butyl ester (150 mg, 0.236 mmol) in DCM (5 mL) was treated with TFA (2 mL) for 2 h. The solvent was removed under vacuum, and the crude product was dissolved in EtOAc, washed with saturated aqueous NaHCO3 solution and brine, dried (MgSO4), and concentrated. The resulting material was purified by reversed-phase chromatography (Gemini NX-C18, 21*150 mm, water (0.1% TFA) / acetonitrile, gradient over 12 min, 25 mL / min). The collected fractions were concentrated to give 6-(4-bromophenylamino)-1-[6-(piperidin-4-yloxy)pyridin-2-yl]-2-(prop-2-enyl)-1,2-dihydro-3-yloxy ...2-(prop-2-enyl)-2-(prop-2-enyl)-2-(prop-2-enyl)-2-(prop-2-enyl)-2-(prop-2-enyl)-2-(prop-2-enyl)-2-(prop-2-enyl)-2-(prop-2-enyl)-2-(prop-2-enyl)-2-(prop-2-enyl)-2-(prop-2-enyl)-2-(prop-2-enyl)-2-(prop-2- H -1,2,5,7-Tetraazaindene-3-one, a white solid. [150 mg].

[0416] 6-(4-Bromophenylamino)-1-[6-(1-methylpiperidin-4-yloxy)pyridin-2-yl]-2-(prop-2-enyl)- 1,2-Dihydro-3H-1,2,5,7-Tetraazaindene-3-one (Compound 370)

[0417] 6-(4-bromophenylamino)-1-[6-(piperidin-4-yloxy)pyridin-2-yl]-2-(prop-2-enyl)-1,2-dihydro-3 H -1,2,5,7-tetraazaindene-3-one (150 mg, 0.236 mmol) was dissolved in anhydrous THF (5 mL). Formaldehyde (approximately 36% pure, 38.3 µL, 0.471 mmol) and STAB (150 mg, 0.707 mmol) were added sequentially. The reaction mixture was stirred at room temperature and monitored by LCMS. After 2 hours, the reaction was quenched dropwise with saturated sodium bicarbonate aqueous solution (5 mL). The aqueous phase was extracted with EtOAc (3 x 20 mL). The combined organic layers were dried (MgSO4) and concentrated under reduced pressure to give a crude material as a yellow powder. The obtained material was purified by reversed-phase chromatography (Gemini NX-C18, 21 x 150 mm, water (0.1% TFA) / acetonitrile, gradient over 12 minutes, 25 mL / min). The purified fractions were collected and concentrated to give the title compound [50.9 mg, yield 33.2%].

[0418] Example 5. Synthesis of 2-ethyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-[m-(5-pyrimidinyl)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 367)

[0419] 4-{6-[6-(3-bromophenylamino)-2-ethyl-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazaindene-1- tert-butyl 1-piperidinecarboxylate (2-pyridyloxy)

[0420] This intermediate is obtained through interaction with 4-{6-[6-(4-bromophenylamino)-3-oxo-2-(prop-2-enyl)-1,2-dihydro-3- H -1,2,5,7-Tetraazaindene-1-yl]pyridin-2-yloxy}piperidine-1-carboxylic acid Uncle Butyl ester is prepared using the same method.

[0421] 2-Ethyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-[m-(5-pyrimidinyl)phenylamino]

[367] -1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one

[0422] To a stirred solution of tert-butyl 4-{6-[6-(3-bromophenylamino)-2-ethyl-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazaindene-1-yl]-2-pyridyloxy}-1-piperidinecarboxylate (230 mg, 376 µmol) in a mixture of 1,4-dioxane (7 mL) and water (2 mL), 2 M K₂CO₃ (600 µl) and hydroxy(5-pyrimidinyl)boronic acid ester (46.6 mg, 376 µmol) were added. The reaction mixture was degassed for 15 minutes. Add bis[2-(diphenylphosphino)-2,4-cyclopentadien-1-ide]-dichloro-palladamethane (33 mg, 0.12 equivalents, 45.1 µmol) and tighten the screw cap onto the sealed tube. Heat the contents to 100 °C and stir overnight. Cool the reaction mixture to room temperature, dilute with EtOAc, wash with water and then with a brine solution. Dry the organic layer with anhydrous Na₂SO₄ and remove the solvent under reduced pressure to obtain the crude material. Dissolve the collected material (150 mg) in DCM:TFA 4:1 v / v. Stir the reaction mixture at room temperature for 1 hour. Concentrate the mixture under vacuum and dissolve the residue in anhydrous THF (5 mL). Formaldehyde (23.4 µL, 2 equivalents, 314 µmol) and sodium triacetoxyborohydride (99.8 mg, 3 equivalents, 471 µmol) were added sequentially. The reaction mixture was stirred at room temperature and monitored by LCMS. After 2 hours, the reaction was quenched dropwise with saturated sodium bicarbonate aqueous solution (5 mL). The aqueous phase was extracted with ethyl acetate (3 x 20 mL). The combined organic layers were decanted through a phase separator and concentrated under reduced pressure to give the product as a brownish-red powder. The obtained material was purified by reversed-phase chromatography (Gemini NX-C18, 21 x 150 mm, water (0.1% TFA) / acetonitrile, gradient over 12 minutes, 25 mL / min). The purified fractions were collected and concentrated to give the title compound.

[0423] Example 6. Synthesis of 2-ethyl-1-[6-(4-piperidinoxy)-2-pyridyl]-6-[m-(5-pyrimidinyl)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 366)

[0424] This compound was prepared using a method similar to that described in the synthesis of 2-ethyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-[m-(5-pyrimidinyl)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0425] Example 7. Synthesis of 2-allyl-6-[m-(1-imidazolyl)phenylamino]-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 365)

[0426] 4-(6-{2-allyl-6-[m-(1-imidazolyl)phenylamino]-3-oxo-1,2-dihydro-3H-1,2,5,7- Tetraazainin-1-yl-2-pyridinyloxy)-1-piperidincarboxylic acid tert-butyl ester

[0427] Under nitrogen atmosphere, at room temperature, 112 mg (assumed 0.493 mmol) of mCPBA (<77% pure) in DCM (0.5 mL) was added to a stirred solution of 4-{6-[2-allyl-6-(methylthio)-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazaindene-1-yl]-2-pyridyloxy}-1-piperidinecarboxylate tert-butyl ester (205 mg, 0.411 mmol) in DCM (5 mL). After 15 minutes, the DCM was evaporated under vacuum, and the crude product was dissolved in MeCN (5 mL). Then, m-(1-imidazolyl)aniline (65.5 mg, 0.411 mmol) and methanesulfonic acid (79 mg, 0.822 mmol) were added. The reaction mixture was stirred in a sealed vial at 60 °C. After 48 hours, the reaction mixture was cooled to room temperature, and mCPBA quenched with 1M NaOH (5 mL) was added dropwise. The aqueous phase was extracted with EtOAc (3 x 20 mL) and washed with brine (20 mL). The combined organic layers were dried (MgSO4) and concentrated under reduced pressure to give the product as a yellow powder. The crude material was dissolved in DCM (5 mL), loaded onto a 10 g silica gel column, and purified by rapid chromatography (0-100%, EtOAc: PE) to give tert-butyl 4-(6-{2-allyl-6-[m-(1-imidazolyl)phenylamino]-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazaindene-1-yl}-2-pyridyloxy)-1-piperidinecarboxylate [160 mg, 62.7%] as a yellow solid.

[0428] 2-Allyl-6-[m-(1-imidazolyl)phenylamino]-1-[6-(4-piperidinyloxy)-2-pyridyl]-1,2- Dihydro-3H-1,2,5,7-tetraazainden-3-one, trifluoroacetate

[0429] A solution of tert-butyl 4-(6-{2-allyl-6-[m-(1-imidazolyl)phenylamino]-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazaindene-1-yl}-2-pyridyloxy)-1-piperidinecarboxylate (160 mg, 0.307 mmol) in DCM (5 mL) was treated with TFA (2 mL) for 2 hours. The solvent was removed under vacuum, and the crude product was dissolved in EtOAc, washed with saturated aqueous NaHCO3 solution and brine, dried (MgSO4), and concentrated. The resulting material was purified by purification phase chromatography (Gemini NX-C18, 21*150 mm, water (0.1% TFA) / acetonitrile, gradient over 12 minutes, 25 mL / min). The pure fractions were collected and concentrated to give 2-allyl-6-[m-(1-imidazolyl)phenylamino]-1-[6-(4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, a trifluoroacetate, as a white solid [150 mg].

[0430] 2-Allyl-6-[m-(1-imidazolyl)phenylamino]-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridine]

[365] -1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one, trifluoroacetate (compound 365)

[0431] 2-Allyl-6-[m-(1-imidazolyl)phenylamino]-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (150 mg, 0.296 mmol) was dissolved in anhydrous THF (5 mL). Formaldehyde (approximately 36% pure, 44 µL, 0.591 mmol) and STAB (188 mg, 0.887 mmol) were added sequentially. The reaction mixture was stirred at room temperature and monitored by LCMS. After 2 hours, the reaction was quenched dropwise with saturated sodium bicarbonate aqueous solution (5 mL). The aqueous phase was extracted with EtOAc (3 x 20 mL). The combined organic layers were dried (MgSO4) and concentrated under reduced pressure to give a crude material as a yellow powder. The obtained material was purified by reversed-phase chromatography (Gemini NX-C18, 21*150 mm, water (0.1% TFA) / acetonitrile, gradient over 12 minutes, 25 ml / min). The purified fractions were collected and concentrated to give 2-allyl-6-[m-(1-imidazolyl)phenylamino]-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one as trifluoroacetate [26.9 mg, yield 28%].

[0432] Example 8. Synthesis of 2-allyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-[m-(1-pyrazolyl)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 364)

[0433] 4-(6-{3-oxo-2-(prop-2-enyl)-6-[3-(pyrazol-1-yl)phenylamino]-1,2-dihydro-3H-1,2, 5,7-Tetraazaindene-1-ylpyridin-2-yloxy)piperidine-1-carboxylic acid tert-butyl ester

[0434] Under nitrogen atmosphere, at room temperature, 114 mg (assumed 0.493 mmol) of mCPBA (<77% pure) in DCM (0.5 mL) was added to a stirred solution of 4-{6-[2-allyl-6-(methylthio)-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazaindene-1-yl]-2-pyridyloxy}-1-piperidinecarboxylate tert-butyl ester (205 mg, 0.411 mmol) in DCM (5 mL). After 15 minutes, the DCM was evaporated under vacuum, and the crude product was dissolved in MeCN (5 mL). Then, m-(1-pyrazolyl)aniline (65.4 mg, 0.411 mmol) and methanesulfonic acid (79 mg, 0.822 mmol) were added. The reaction mixture was stirred in a sealed vial at 60 °C. After 48 hours, the reaction mixture was cooled to room temperature, and mCPBA quenched with 1M NaOH (5 mL) was added dropwise. The aqueous phase was extracted with EtOAc (3 x 20 mL) and then with 20 mL of brine. The combined organic layers were dried (MgSO4) and concentrated under reduced pressure to give the product as a yellow powder. The crude material was dissolved in DCM (5 mL), loaded onto a 10 g silica gel column, and purified by rapid chromatography (0-100%, EtOAc: PE) to give tert-butyl 4-(6-{3-oxo-2-(prop-2-enyl)-6-[3-(pyrazol-1-yl)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-1-yl}pyridin-2-yloxy)piperidine-1-carboxylic acid [140 mg, 58.5%] as a white solid.

[0435] 2-Allyl-1-[6-(4-piperidinyloxy)-2-pyridyl]-6-[m-(1-pyrazolyl)phenylamino]-1,2- Dihydro-3H-1,2,5,7-tetraazainden-3-one, trifluoroacetate

[0436] 4-(6-{3-oxo-2-(prop-2-enyl)-6-[3-(pyrazol-1-yl)phenylamino]-1,2-dihydro-3- H -1,2,5,7-Tetraazaindene-1-ylpyridin-2-yloxy)piperidine-1-carboxylic acid UncleA solution of butyl ester (140 mg, 0.275 mmol) in DCM (5 mL) was treated with TFA (2 mL) for 2 hours. The solvent was removed under vacuum, and the crude product was dissolved in EtOAc, washed with saturated aqueous solution of NaHCO3 and brine, dried (MgSO4), and concentrated. The resulting material was purified by reversed-phase chromatography (GeminiNX-C18, 21*150 mm, water (0.1% TFA) / acetonitrile, gradient over 12 minutes, 25 mL / min). The purified fractions were collected and concentrated to give 2-allyl-1-[6-(4-piperidinyloxy)-2-pyridyl]-6-[m-(1-pyrazolyl)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, as trifluoroacetate, as a white solid [140 mg].

[0437] 2-Allyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-[m-(1-pyrazolyl)phenylamino]

[364] -1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one

[0438] 2-Allyl-1-[6-(4-piperidinoxy)-2-pyridyl]-6-[m-(1-pyrazolyl)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (140 mg, 0.275 mmol) was dissolved in anhydrous THF (5 mL). Formaldehyde (approximately 36% pure, 40.9 µL, 0.549 mmol) and STAB (175 mg, 0.824 mmol) were added sequentially. The reaction mixture was stirred at room temperature and monitored by LCMS. After 2 hours, the reaction was quenched dropwise with saturated sodium bicarbonate aqueous solution (5 mL). The aqueous phase was extracted with EtOAc (3 x 20 mL). The combined organic layers were dried (MgSO4) and concentrated under reduced pressure to give a crude material as a yellow powder. The obtained material was purified by reversed-phase chromatography (Gemini NX-C18, 21*150 mm, water (0.1% TFA) / acetonitrile, gradient over 12 minutes, 25 ml / min). The purified fractions were collected and concentrated to give 2-allyl-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-6-[m-(1-pyrazolyl)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one [25 mg, yield 27%].

[0439] Example 9. Synthesis of 2-allyl-1-[6-(4-piperidinoxy)-2-pyridyl]-6-[m-(1-pyrazolyl)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 363)

[0440] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-[m-(1-pyrazolyl)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0441] Example 10. 2-Allyl-6-(1-Isopropyl-1H-indazole-5-ylamino)-1-(6-{1-[( 2 Synthesis of [H3]-4-piperidinoxy[-2-pyridinyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 354)

[0442] At room temperature, methanol-d4 (31 µl, 0.76 mmol, 4.0 equivalents) and water (2 µl, 0.11 mmol, 0.58 equivalents) were added to a suspension of Dess-Martin periodinane (129 mg, 0.30 mmol, 1.6 equivalents) in DCM (2 ml), and the mixture was stirred for 20 minutes before being filtered through a 0.45 µm syringe filter. The resulting clear solution of formaldehyde-d2 (1.6 equivalents) was added to a solution of 1-[6-(piperidin-4-yloxy)pyridin-2-yl]-2-(prop-2-en-1-yl)-6-{[1-(propan-2-yl)-1H-indazol-5-yl]amino}-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (100 mg, 0.19 mmol, 1.0 equivalents) in THF (3 ml), followed by the addition of sodium triacetoxyborodeuteride (81 mg, 0.38 mmol, 2.0 equivalents), and the mixture was stirred at room temperature. After 1 hour, a second portion of formaldehyde-d2 (0.8 equivalents) prepared in the same manner as described above was added, followed by the addition of sodium triacetoxyborodeuteride (50 mg, 0.23 mmol, 1.2 equivalents). One hour later, LCMS indicated complete conversion of the secondary amine. The mixture was diluted with EtOAc (30 ml), washed with NaCl (2 x 30 ml of 15% aqueous solution), adjusted to pH 11 with NaOH aqueous solution and then with saturated brine (30 ml), filtered through a phase separator, and concentrated under reduced pressure. The crude material was purified by reversed-phase chromatography (Gemini NX-C18, 21 x 150 mm, water (0.1% TFA) / acetonitrile, gradient over 12 minutes, 25 ml / min) and the purified fraction was lyophilized to give the title compound. Yield: 91 mg TFA salt (73%), as a pale yellow powder.

[0443] Example 11. Synthesis of 6-(4-biphenylamino)-2-ethyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 284)

[0444] This compound was prepared using a method similar to that described in the synthesis of 2-ethyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-[m-(5-pyrimidinyl)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0445] Example 12. Synthesis of 2-ethyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-[m-(3-pyridyl)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 352)

[0446] This compound was prepared using a method similar to that described in the synthesis of 2-ethyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-[m-(5-pyrimidinyl)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0447] Example 13. Synthesis of 2-ethyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-[p-(1-methyl-4-pyrazolyl)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 277)

[0448] This compound was prepared using a method similar to that described in the synthesis of 2-ethyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-[m-(5-pyrimidinyl)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0449] Example 14. Synthesis of 6-(4-biphenylamino)-2-ethyl-1-[6-(4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 272)

[0450] This compound was prepared using a method similar to that described in the synthesis of 2-ethyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-[m-(5-pyrimidinyl)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0451] Example 15. Synthesis of 2-ethyl-6-[p-(1-methyl-4-pyrazolyl)phenylamino]-1-[6-(4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 271)

[0452] This compound was prepared using a method similar to that described in the synthesis of 2-ethyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-[m-(5-pyrimidinyl)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0453] Example 16. Synthesis of 2-allyl-6-(m-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 269)

[0454] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0455] Example 17. Synthesis of 6-(p-bromophenylamino)-2-ethyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 268)

[0456] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0457] Example 18. Synthesis of 6-(p-bromophenylamino)-2-ethyl-1-[6-(4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 267)

[0458] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0459] Example 19. Synthesis of 2-allyl-6-(1-isopropyl-1H-indazol-5-ylamino)-1-[6-(4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 265)

[0460] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, yielding a crude free base intermediate (282 mg) as a brown solid, of which 9% was purified by preparative HPLC. Yield: 20 mg TFA salt (64%), as a pale yellow powder.

[0461] Example 20. Synthesis of 2-allyl-6-(2-methoxy-4-pyridinylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 263)

[0462] 4-(3-(2-allyl-6-amino-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy tert-butyl piperidine-1-carboxylate

[0463] Under an inert atmosphere and at room temperature, 400 mg (0.755 mmol) of 4-(3-(2-allyl-6-(methanesulfonyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylic acid tert-butyl ester was placed in a flask, and 5 ml (5.00 mmol) of ammonia in THF solution was added. The mixture was stirred for 16 hours. The reaction progress was monitored by TLC and LCMS. After the reaction was completed, the solvent was evaporated under reduced pressure to obtain the crude compound, which was purified by column chromatography (100-200 mesh silica, eluent of 80-100% ethyl acetate and hexane) to obtain pure compound 4-(3-(2-allyl-6-amino-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylic acid tert-butyl ester (260 mg, 0.346 mmol, 45.7% yield), as a brown solid.

[0464] 2-Allyl-6-(2-methoxy-4-pyridinylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridine]

[263] -1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one

[0465] At room temperature, CuI (1.2 equivalents) followed by N,N'-dimethylethylenediamine (1 equivalent) was added sequentially to a stirred, degassed suspension of 4-({6-[6-amino-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]pyridin-2-yl}oxy)piperidin-1-carboxylic acid tert-butyl ester (200 mg), 4-bromo-2-methoxypyridine (1 equivalent), and cesium carbonate (3 equivalents) in dioxane. The reaction was heated to 90°C overnight in a sealed vial. The reaction was monitored by LCMS, and additional CuI and ligands were added as needed to achieve complete conversion. The reaction mixture was diluted with water and a few drops of ammonia solution (28%) and then extracted with ethyl acetate (×3). The combined organic layers were dried using a phase separator and concentrated under reduced pressure to give an oil. This substance was used without further purification.

[0466] At room temperature, trifluoroacetic acid (10-20% by volume) was added to a stirred solution of the above material (1 equivalent) in anhydrous dichloromethane (0.4 mol / L). The resulting solution was stirred until the Boc-amine intermediate was consumed (15 min, LCMS), and then partitioned between EtOAc and saturated brine, adjusting the pH to approximately 12 with aqueous NaOH solution. The organic phase was filtered through a phase separator and concentrated under reduced pressure to give a crude intermediate as a red solid. A portion of this material was purified by reversed-phase chromatography (Gemini NX-C18, 21*150 mm, water (0.1% TFA) / acetonitrile, gradient within 12 min, 25 ml / min), and the purified fraction was lyophilized to give the amine compound as a TFA salt.

[0467] The material was methylated using a method similar to that described for 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one. Yield: 178 mg TFA salt (71%), as a pale yellow powder.

[0468] Example 21. Synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[2-(4-piperidinyloxy)-4-pyrimidinyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 350)

[0469] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridinylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 157 mg TFA salt (62%), as a yellow powder.

[0470] Example 22. Synthesis of 2-allyl-6-(2-methyl-1,3-benzothiazol-6-ylamino)-1-[6-(4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 264)

[0471] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridinylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 15 mg TFA salt (61%), as a yellow powder.

[0472] Example 23. Synthesis of 2-ethyl-6-[m-(1-methyl-4-pyrazolyl)phenylamino]-1-[6-(4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 340)

[0473] This compound was prepared using a method similar to that described in the synthesis of 2-ethyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-[m-(5-pyrimidinyl)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0474] Example 24. Synthesis of 6-(3-Biphenylamino)-2-ethyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 339)

[0475] This compound was prepared using a method similar to that described in the synthesis of 2-ethyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-[m-(5-pyrimidinyl)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0476] Example 25. Synthesis of 6-(3-biphenylamino)-2-ethyl-1-[6-(4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 338)

[0477] This compound was prepared using a method similar to that described in the synthesis of 2-ethyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-[m-(5-pyrimidinyl)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0478] Example 26. Synthesis of 2-allyl-6-(2-methyl-1,3-benzothiazol-6-ylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 260)

[0479] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridinylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 65 mg TFA salt (62%), as a pale yellow powder.

[0480] Example 27. Synthesis of 2-allyl-6-(6-methoxy-3-pyridylamino)-1-[6-(4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 251)

[0481] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0482] Example 28. Synthesis of 2-allyl-6-(6-methoxy-3-pyridylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 250)

[0483] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0484] Example 29. Synthesis of 2-allyl-6-(1,3,3a-triaza-5-indenylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (compound 249)

[0485] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 26 mg TFA salt (56%), as a pale yellow powder.

[0486] Example 30. Synthesis of 2-allyl-6-(1-isopropyl-1H-indazol-5-ylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 248)

[0487] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 260 mg TFA salt (81%), as a pale yellow powder.

[0488] Example 31. Synthesis of 2-allyl-6-(1,3,3a-triaza-5-indenylamino)-1-[6-(4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (compound 245)

[0489] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 53 mg TFA salt (22%), as a yellow powder.

[0490] Example 32. Synthesis of 2-allyl-6-(2,1,3-benzothiadiazol-5-ylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 208)

[0491] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 58 mg TFA salt (64%), as a pale yellow powder.

[0492] Example 33. Synthesis of 2-allyl-6-(6-isoquinolinylamino)-1-[6-(4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 244)

[0493] 4-{6-[2-allyl-6-(6-isoquinolinylamino)-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraaza-} Indene-1-yl]-2-pyridyloxy}-1-piperidinecarboxylic acid tert-butyl ester

[0494] Under nitrogen atmosphere, tert-butyl 4-[6-(2-allyl-6-amino-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazaindene-1-yl)-2-pyridyloxy]-1-piperidinecarboxylate (100 mg, 0.214 mmol), 6-bromoisoquinoline (46.4 mg, 0.214 mmol), and Cs₂CO₃ (209 mg, 3 equivalents) were mixed in dioxane (3 mL) and stirred for 15 min. Catalytic amounts of CuI (48.9 mg, 1.2 equivalents) and 1,2-bis(methylamino)ethane as ligands (18.9 mg, 0.214 mmol) were added. The reaction mixture was heated to 90 °C and maintained for 18 h. The reaction progress was observed by LCMS. The reaction mixture was cooled to room temperature, and the crude residue was quenched with saturated ammonia solution to remove CuI. The mixture was then extracted with EtOAc (3 × 10 mL) and brine. The combined organic layers were dried (anhydrous Na₂SO₄) and the solvent was evaporated under reduced pressure. The crude product was dissolved in MeCN (2 mL) and crystallized over 1 hour by slow addition of water (2 mL). This yielded tert-butyl 4-{6-[2-allyl-6-(6-isoquinolinylamino)-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazaindene-1-yl]-2-pyridyloxy}-1-piperidinecarboxylate [33 mg, 30% yield], as a grayish-white solid. LCMS (ESI+), m / z: 596.

[0495] 2-Allyl-6-(6-isoquinolinylamino)-1-[6-(4-piperidinyloxy)-2-pyridyl]-1,2-dihydro- 3H-1,2,5,7-Tetraazainden-3-one (Compound 244)

[0496] A solution of tert-butyl 4-{6-[2-allyl-6-(6-isoquinolinylamino)-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazaindene-1-yl]-2-pyridyloxy}-1-piperidinecarboxylate (33 mg, 0.055 mmol) in DCM (2 mL) was treated with TFA (0.5 mL) for 2 hours. The solvent was removed under vacuum, and the crude product was dissolved in EtOAc, washed with saturated aqueous solution of NaHCO3 and brine, dried (MgSO4), and concentrated. The resulting material was purified by crystallization over 1 hour with the slow addition of water (2 mL) in MeCN (2 mL). This yields 2-allyl-6-(6-isoquinolinylamino)-1-[6-(4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, a yellow solid [30 mg].

[0497] Example 34. Synthesis of 2-allyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-(7-quinolinylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 335)

[0498] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0499] Example 35. Synthesis of 2-allyl-1-[6-(4-piperidinyloxy)-2-pyridyl]-6-(6-quinolinylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 242)

[0500] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0501] Example 36. Synthesis of 2-allyl-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-6-(6-quinoxalinylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 241)

[0502] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0503] Example 37. Synthesis of 2-allyl-6-(7-isoquinolinylamino)-1-[6-(4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 240)

[0504] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0505] Example 38. Synthesis of 2-allyl-1-[6-(4-piperidinoxy)-2-pyridyl]-6-(6-quinoxalinylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 239)

[0506] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0507] Example 39. Synthesis of 1-{6-[(S)-1-methyl-3-piperidinyloxy]-2-pyridyl}-2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 332)

[0508] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 61 mg TFA salt (82%), as a pale yellow powder.

[0509] Example 40. Synthesis of 2-allyl-6-(5-fluoro-3-pyridinylamino)-1-[6-(4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 329)

[0510] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridinylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 157 mg TFA salt (62%), as a yellow powder. Yield: 10 mg TFA salt (46%).

[0511] Example 41. Synthesis of 6-(1,3a-diaza-5-indenylamino)-2-allyl-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (compound 328)

[0512] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 70 mg TFA salt (80%), as a yellow solid.

[0513] Example 42. Synthesis of 2-allyl-6-(5-fluoro-3-pyridinylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 327)

[0514] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridinylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 157 mg TFA salt (62%), as a yellow powder. Yield: 125 mg TFA salt (83%), as a yellow solid.

[0515] Example 43. Synthesis of 2-allyl-6-(7-isoquinolinylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 326)

[0516] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0517] Example 44. Synthesis of 6-(1,3a-diaza-5-indenylamino)-2-allyl-1-[6-(4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (compound 324)

[0518] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 100 mg TFA salt (42%).

[0519] Example 45. Synthesis of 2-allyl-6-(5-chloro-3-pyridinylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 323)

[0520] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0521] Example 46. Synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[6-(1-propyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 322)

[0522] 2-Allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[6-(4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(piperidin-4-yloxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one.

[0523] Propanol (37.7 µL, 0.502 mmol) was added to a suspension of Desmond-Martin periodoyl ether (63.9 mg, 0.151 mmol) in DCM (4 mL), followed by the addition of 9 mL of H₂O. The mixture was stirred for 30 minutes until it became a white suspension, and then filtered. At room temperature, 2 mL of the filtered solution was added to a stirred solution of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[6-(4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (50 mg, 0.1 mmol) in THF. STAB (63.9 mg, 0.301 mmol) was added. The mixture was stirred overnight at room temperature. The mixture was diluted with EtOAc (20 ml) and washed with a brine mixture, which was then slightly alkaline with NaOH (2 x 20 ml). The organic phase was dried through a phase separator and concentrated under vacuum. The crude material was purified by reversed-phase chromatography (Gemini NX-C18, 21 x 150 mm, water (0.1% TFA) / acetonitrile, gradient over 12 minutes, 25 ml / min). The purified fractions were collected and concentrated to give 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[6-(1-propyl-4-piperidinyloxy)-2-pyridinyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, as trifluoroacetate (26 mg).

[0524] Example 47. Synthesis of 2-allyl-1-[6-(1-ethyl-4-piperidinyloxy)-2-pyridyl]-6-(1-methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 321)

[0525] EtOH (29.3 µL, 0.502 mmol) was added to a suspension of Desmond-Martin periodoylene (63.9 mg, 0.151 mmol) in DCM (4 mL), followed by 9 mL of H₂O. The mixture was stirred for 30 minutes until a suspension was formed, and then filtered. At room temperature, 2 mL of the filtered solution was added to a stirred solution of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[6-(4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (50 mg, 0.1 mmol) in THF. STAB (42.6 mg, 0.201 mmol) was added. The mixture was stirred overnight at room temperature. The mixture was diluted with EtOAc (20 ml) and washed with a brine mixture, which was then slightly alkaline with NaOH (2 x 20 ml). The organic phase was dried through a phase separator and concentrated under vacuum. The crude material was purified by reversed-phase chromatography (Gemini NX-C18, 21 x 150 mm, water (0.1% TFA) / acetonitrile, gradient over 12 minutes, 25 ml / min). The purified fractions were collected and concentrated to give 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[6-(1-propyl-4-piperidinyloxy)-2-pyridinyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, as trifluoroacetate (25 mg).

[0526] Example 48. Synthesis of 2-allyl-6-(1,2-benzisothiazo-5-ylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 320)

[0527] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 58 mg TFA salt (64%), as a pale yellow powder.

[0528] Example 49. Synthesis of 2-allyl-1-[m-(1-methyl-4-piperidinoxy)phenyl]-6-(2-methyl-4-pyridylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 219)

[0529] Formaldehyde (310 mg, 3.82 mmol) was added to a stirred solution of 2-allyl-6-((2-methylpyridin-4-yl)amino)-1-(3-(piperidin-4-yloxy)phenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (350 mg, 0.765 mmol) in THF (5 mL), and the reaction mixture was stirred at 25 °C for 5 min. STAB (486 mg, 2.295 mmol) was then added in portions. After complete addition of STAB, the reaction mixture was stirred at 25 °C for 20 min. The progress of the reaction was monitored by TLC and LCMS. The reaction mixture was then quenched with TFA, followed by quenching with MeOH solution of NH3, diluted with water, and extracted with 10% MeOH in DCM solution (2 x 100 mL). The combined organic extracts were washed with NaHCO3, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain a crude compound. This crude compound was purified by preparative HPLC to obtain pure compound 2-allyl-1-(3-((1-methylpiperidin-4-yl)oxy)phenyl)-6-((2-methylpyridin-4-yl)amino)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (66 mg, 0.136 mmol, 17.75% yield), which was a grayish-white solid.

[0530] Example 50. Synthesis of 2-allyl-6-(p-fluorophenylamino)-1-[6-(1-methyl-4-azacycloheptyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 317)

[0531] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0532] Example 51. Synthesis of 2-allyl-6-(2-methyl-1,3-benzoxazol-5-ylamino)-1-[6-(4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 232)

[0533] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 13.5 mg TFA salt (50%), as a yellow powder.

[0534] Example 52. Synthesis of 2-allyl-6-(2-methyl-1,3-benzoxazol-6-ylamino)-1-[6-(4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 316)

[0535] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 17.6 mg TFA salt (65%), as a yellow powder.

[0536] Example 53. Synthesis of 2-allyl-6-(2-methyl-1,3-benzoxazol-5-ylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 231)

[0537] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 19 mg TFA salt (80%), as a pale yellow powder.

[0538] Example 54. Synthesis of 2-allyl-6-(2-methyl-4-pyridylamino)-1-[m-(4-piperidinoxy)phenyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 222)

[0539] 4-(3-bromophenoxy)piperidine-1-carboxylic acid tert-butyl ester

[0540] Under an inert atmosphere, NaH (2.285 g, 57.1 mmol) was added to a stirred solution of tert-butyl 4-hydroxypiperidine-1-carboxylate (7.48 g, 37.1 mmol) in DMF (50 ml) at 0 °C, and the mixture was stirred at 50 °C for 1 hour. Subsequently, the flask was cooled to room temperature, and 1-bromo-3-fluorobenzenefluorobenzene (5 g, 28.6 mmol) dissolved in DMF (10 ml) was added to the reactants and stirred at 70 °C for 3 hours. The reaction progress was monitored by LCMS and TLC. After the reaction was complete, the reactants were quenched with ice-cold water and extracted with EtOAc (500 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude compound. This crude compound was purified by column chromatography (100-200 mesh silica, eluent: 10-20% ethyl acetate and hexane) to give tert-butyl 4-(3-bromophenoxy)piperidine-1-carboxylic acid (6.8 g, 14.89 mmol, 52.1% yield), a yellow gelatinous liquid. LCMS m / z measured value: 300.0 (M-56).

[0541] N-(6-(2-allyl-6-(methylthio)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl) pyridin-2-yl)-2-cyano-N-methylacetamide

[0542] Under an inert atmosphere, a stirred solution of 2-allyl-6-(methylthio)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (420 mg, 1.890 mmol), tert-butyl 4-(3-bromophenoxy)piperidin-1-carboxylate (808 mg, 2.268 mmol), K₂CO₃ (783 mg, 5.67 mmol), and N,N′-dimethylethylenediamine (0.203 mL, 1.890 mmol) in dioxane (5 mL) was degassed for 20 min at room temperature. CuI (359 mg, 1.890 mmol) was added, and the mixture was degassed again for 5 min, followed by stirring at 110 °C for 16 h. The reaction progress was monitored by TLC and LCMS. After the reaction was complete, the reaction mixture was filtered through diatomaceous earth (celite) and washed with 10% MeOH in DCM solution (150 mL). The collected fractions were concentrated under reduced pressure to obtain a crude compound, which was then purified by rapid column chromatography (SiO2 / 230-400 mesh; 20-50% ethyl acetate-petroleum ether) to give tert-butyl 4-(3-(2-allyl-6-(methylthio)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylic acid (65 mg, 0.108 mmol, 5.74% yield), as a grayish-white solid.

[0543] tert-butyl-4-(3-(2-allyl-6-(methanesulfonyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]) Pyrimidin-1-yl)phenoxy)piperidine-1-carboxylate

[0544] m-CPBA (41.6 mg, 0.241 mmol) was added to a stirred solution of 4-(3-(2-allyl-6-(methylthio)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylic acid tert-butyl ester (60 mg, 0.121 mmol) in DCM (2 ml). The mixture was stirred at room temperature for 2 hours. The reaction progress was monitored by TLC and LCMS. After the reaction was complete, the reaction mixture was quenched with sodium bicarbonate solution and then extracted with DCM solution of 10% MeOH. The organic layer was dried over sodium sulfate and concentrated under reduced pressure to give crude tert-butyl-4-(3-(2-allyl-6-(methanesulfonyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylate (60 mg, 0.113 mmol, 94% yield). This crude compound was used in the next step without any further purification.

[0545] tert-butyl-4-(3-(2-allyl-6-amino-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidine-1- 1-Phenoxy-piperidine-1-carboxylate

[0546] Under an inert atmosphere, at room temperature, 400 mg (0.755 mmol) of 4-(3-(2-allyl-6-(methanesulfonyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylic acid tert-butyl ester was added to a flask, followed by the addition of 5 mL (5.00 mmol) of ammonia in THF, and the mixture was stirred for 16 hours. The reaction progress was monitored by TLC and LCMS. After the reaction was completed, the solvent was evaporated under reduced pressure to obtain the crude compound, which was purified by column chromatography (100-200 mesh silica, eluent of 80-100% ethyl acetate and hexane) to obtain tert-butyl 4-(3-(2-allyl-6-amino-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylic acid (260 mg, 0.346 mmol, 45.7% yield), as a brown solid.

[0547] tert-butyl-4-(3-(2-allyl-6-((2-methylpyridin-4-yl)amino)-3-oxo-2,3-dihydro-1H-pyridine) Azo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylate

[0548] A stirred solution of 4-(3-(2-allyl-6-amino-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylic acid tert-butyl ester (300 mg, 0.643 mmol), 4-bromo-2-methylpyridine (133 mg, 0.772 mmol), K₂CO₃ (267 mg, 1.929 mmol), and N,N'-dimethylethylenediamine (56.6 mg, 0.643 mmol) in dioxane (4 mL) was degassed with N₂ for 30 min. Copper(I) iodide (122 mg, 0.643 mmol) was added to the mixture, and the mixture was degassed again for 5 min. The mixture was stirred at 110 °C for 16 h. The reaction progress was monitored by TLC and LCMS. After the reaction was complete, the reactants were filtered through diatomaceous earth, and the crude compound was washed with ethyl acetate (100 mL). The combined organic layers were dried over anhydrous Na₂SO₄ and concentrated under vacuum to obtain a crude compound. This crude compound was purified by column chromatography (100-200 mesh silica, eluent 80-100% ethyl acetate and hexane) to give tert-butyl 4-(3-(2-allyl-6-((2-methylpyridin-4-yl)amino)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylic acid (140 mg, 0.208 mmol, 32.4% yield), as a grayish-white solid. LCMS m / z: 558.5 (M+H).

[0549] 2-Allyl-6-((2-methylpyridin-4-yl)amino)-1-(3-(piperidin-4-yloxy)phenyl)-1,2-di Hydrogen-3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 222)

[0550] Under an inert atmosphere and at 0 °C, a solution of HCl in dioxane (0.2 ml, 0.800 mmol) was added to a stirred solution of 4-(3-(2-allyl-6-((2-methylpyridin-4-yl)amino)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylic acid tert-butyl ester (40 mg, 0.071 mmol) in dioxane (1 ml). The mixture was then stirred at room temperature for 16 hours. The reaction progress was monitored by TLC and LCMS. After the reaction was complete, the solvent was evaporated under reduced pressure to obtain the crude compound, which was washed with n-hexane. The crude compound was then purified by preparative HPLC (column: X-select CSH C18, mobile phase A: water (containing 0.1% AA), mobile phase B: acetonitrile, flow rate: 15.0 mL / min, Rt: 12.8) to give 2-allyl-6-((2-methylpyridin-4-yl)amino)-1-(3-(piperidin-4-yloxy)phenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (14 mg, 0.030 mmol, 42.65% yield).

[0551] Example 55. Synthesis of 2-allyl-1-[6-(4-piperidinoxy)-2-pyridyl]-6-(3-pyridylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 230)

[0552] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridinylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 93 mg, as a white powder.

[0553] Example 56. Synthesis of 2-allyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-(3-pyridylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 229)

[0554] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridinylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 96 mg TFA salt (87%), as a pale yellow powder.

[0555] Example 57. Synthesis of 2-allyl-1-[6-(4-azacycloheptyloxy)-2-pyridyl]-6-(p-fluorophenylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 315)

[0556] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0557] Example 58. Synthesis of 2-allyl-6-(1,2-benzisothiazol-5-ylamino)-1-[6-(4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 228)

[0558] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0559] Example 59. Synthesis of 2-allyl-1-[2-(1-methyl-4-piperidinylamino)-4-pyrimidinyl]-6-[p-(2,2,2-trifluoroethoxy)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 314)

[0560] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one. Yield: 49 mg TFA salt (57%), as a white powder.

[0561] Example 60. Synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[2-(1-methyl-4-piperidinylamino)-4-pyrimidinyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 313)

[0562] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one. Yield: 24.5 mg TFA salt (40%), as a yellow powder.

[0563] Example 61. Synthesis of 2-allyl-6-(p-fluorophenylamino)-1-[2-(1-methyl-4-piperidinylamino)-4-pyrimidinyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 312)

[0564] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one. Yield: 40 mg TFA salt (54%), as a white powder.

[0565] Example 62. Synthesis of 2-allyl-1-[m-(1-methyl-4-piperidinoxy)phenyl]-6-(1-methyl-4-pyrazolylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 221)

[0566] 4-(3-(2-allyl-6-((1-methyl-1H-pyrazol-4-yl)amino)-3-oxo-2,3-dihydro-1H-pyrazol) [3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylic acid tert-butyl ester

[0567] At 25 °C, 1-methyl-1H-pyrazol-4-amine (14.67 mg, 0.151 mmol) was added to a stirred solution of 4-(3-(2-allyl-6-(methanesulfonyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylic acid tert-butyl ester (80 mg, 0.151 mmol) in acetic acid (3 mL), and the reaction mixture was stirred at 25 °C for 16 h. The reaction progress was monitored by TLC and LCMS. After the reaction was complete, the reaction mixture was concentrated under reduced pressure, diluted with 10% aqueous sodium bicarbonate solution, and extracted with 10% MeOH in DCM solution. The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude compound. This crude compound was purified by column chromatography (100-200 mesh silica gel, eluent 5-10% MeOH in DCM solution) to give tert-butyl 4-(3-(2-allyl-6-((1-methyl-1H-pyrazol-4-yl)amino)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylic acid (60 mg, 0.082 mmol, 54.5% yield), as a grayish-white solid. LCMS m / z: 547.6 (M+1).

[0568] 2-Allyl-6-((1-methyl-1H-pyrazol-4-yl)amino)-1-(3-(piperidin-4-yloxy)phenyl)-1,2- Dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one

[0569] Under an inert atmosphere and at 0 °C, a solution of 4 M HCl (0.329 mL, 1.317 mmol) in dioxane was added to a stirred solution of 60 mg (0.110 mmol) of 4-(3-(2-allyl-6-((1-methyl-1H-pyrazol-4-yl)amino)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylic acid in 1,4-dioxane (5 mL). The reaction mixture was then stirred at room temperature for 8 hours. The reaction progress was monitored by TLC and LCMS. After the reaction was completed, the reactants were concentrated under reduced pressure to obtain a crude compound, which was then purified by preparative HPLC (column: X-select CSH C18, mobile phase A: water (containing 0.1% FA), mobile phase B: acetonitrile (containing 0.1% FA), flow rate: 15.0 mL / min, Rt: 12.8) to obtain 2-allyl-6-((1-methyl-1H-pyrazol-4-yl)amino)-1-(3-(piperidin-4-yloxy)phenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (8 mg, 0.018 mmol, 16% yield), as a brown solid.

[0570] 2-Allyl-1-[m-(1-methyl-4-piperidinoxy)phenyl]-6-(1-methyl-4-pyrazolylamino)-1, 2-Dihydro-3H-1,2,5,7-Tetraazainden-3-one (Compound 221)

[0571] At 25 °C, 37% formaldehyde aqueous solution (0.083 mL, 1.120 mmol) was added to a stirred solution of 2-allyl-6-((1-methyl-1H-pyrazol-4-yl)amino)-1-(3-(piperidin-4-yloxy)phenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (100 mg, 0.224 mmol) in THF (5 mL), and the mixture was stirred for 10 min. STAB (142 mg, 0.672 mmol) was then added in portions. After complete addition of STAB, the reaction mixture was stirred at 25 °C for 3 h. The reaction progress was monitored by UPLC. After the reaction was complete, the reaction mixture was quenched with TFA, followed by ammonia in MeOH solution. The reaction mixture was diluted with water and extracted with 10% MeOH in DCM solution. The combined organic extracts were washed with an aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude compound. This crude compound was purified by preparative HPLC (column: X-select CSH C18, mobile phase A: aqueous solution of ammonium acetate, mobile phase B: acetonitrile, flow rate: 15.0 mL / min, Rt-10.8) to give 2-allyl-6-((1-methyl-1H-pyrazol-4-yl)amino)-1-(3-((1-methylpiperidin-4-yl)oxy)phenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one.

[0572] Example 63. Synthesis of 2-allyl-6-(2-methyl-1,3-benzoxazol-6-ylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 311)

[0573] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 184 mg TFA salt (84%), as a pale yellow powder.

[0574] Example 64. Synthesis of 1-{m-[N-methyl(1-methyl-4-piperidinyl)amino]phenyl}-2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 225)

[0575] 2-Allyl-6-(methylthio)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one

[0576] At room temperature, 3-nitrophenylboronic acid (1.352 g, 8.10 mmol), sodium carbonate (1.707 g, 16.20 mmol), and copper(II) acetate (0.49 g, 2.70 mmol) were added to a stirred solution of 2-allyl-6-(methylthio)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (1.2 g, 5.40 mmol) in dichloromethane (15 mL), followed by pyridine (0.169 g, 1.080 mmol). The mixture was heated to 70 °C and stirred for 16 hours. The reaction progress was monitored by TLC and LCMS. After the reaction was complete, the mixture was filtered through diatomaceous earth and washed with 10% MeOH in DCM solution (2 x 100 mL). The combined organic layers were dried over anhydrous Na₂SO₄ and concentrated under reduced pressure to obtain a crude compound. This crude compound was purified by column chromatography (100-200 mesh silica, eluent 70-100% ethyl acetate and hexane) to give 2-allyl-6-(methylthio)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (700 mg, 1.794 mmol, 33.2% yield) as a white solid. LCMS m / z: 344.2 (M+H).

[0577] 2-Allyl-6-(Methylsulfonyl)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidine- 3-keto

[0578] Under an inert atmosphere, at room temperature, 2-allyl-6-(methylthio)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (50 mg, 0.146 mmol) was added to a stirred solution of 2-allyl-6-(methylthio)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (10 ml) in DCM. m CPBA (53.7 mg, 0.218 mmol). The mixture was stirred for 2 hours. The reaction progress was monitored by TLC and UPLC. After the reaction was complete, the reaction mixture was quenched with 10% sodium bicarbonate aqueous solution (10 mL) and extracted with 10% MeOH in DCM solution (2 x 50 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude compound 2-allyl-6-(methanesulfonyl)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (30 mg, 0.024 mmol, 16.47% yield) as a grayish-white solid. This crude compound was used for the next step without further purification.

[0579] 2-Allyl-6-((1-Methyl-1H-indazol-5-yl)amino)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyridine Azo[3,4-d]pyrimidin-3-one

[0580] At room temperature, 1-methyl-1H-indazole-5-amine (0.3 g, 2.039 mmol) was added to a stirred solution of 2-allyl-6-(methanesulfonyl)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (0.7 g, 2.039 mmol) in AcOH (5 mL) and stirred for 16 hours. The reaction progress was monitored by TLC and UPLC. After the reaction was complete, the solvent was evaporated under reduced pressure, and the resulting residue was quenched with 10% sodium bicarbonate aqueous solution (100 mL). The resulting mixture was extracted with ethyl acetate (2 x 300 mL). The combined organic extracts were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude compound. This crude compound was purified by column chromatography (100-200 mesh silica, eluent 10-20% ethyl acetate and hexane) to give 2-allyl-6-((1-methyl-1H-indazol-5-yl)amino)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (600 mg, 1.221 mmol, 66.51% yield) as a yellow solid.

[0581] (2-Allyl-1-(3-nitrophenyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-6-yl) (1-Methyl-1H-indazol-5-yl)tert-butyl carbamate

[0582] TEA (0.452 mmol) and Boc anhydride (0.148 g, 0.678 mmol) were added to a stirred solution of 2-allyl-6-((1-methyl-1H-indazol-5-yl)amino)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (0.2 g, 0.452 mmol) in DCM (5 mL). The mixture was stirred at room temperature under an inert atmosphere for 16 hours. The reaction progress was monitored by TLC and UPLC. After the reaction was complete, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (2 x 200 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound (2-allyl-1-(3-nitrophenyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-6-yl)(1-methyl-1H-indazol-5-yl)carbamate tert-butyl ester (250 mg, 0.424 mmol, 94% yield), as a white solid. LCMS m / z: 443.4 (M-100).

[0583] (2-Allyl-1-(3-aminophenyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-6-yl) (1-Methyl-1H-indazol-5-yl)tert-butyl carbamate

[0584] Iron (257 mg, 4.61 mmol) and NH₄Cl (246 mg, 4.61 mmol) were added to a stirred solution of (2-allyl-1-(3-nitrophenyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-6-yl)(1-methyl-1H-indazol-5-yl)carbamate (250 mg, 0.461 mmol) in EtOH (3.0 mL) and water (10 mL), and the mixture was stirred for 16 hours. The reaction progress was monitored by TLC and UPLC. After the reaction was complete, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (2 x 30 mL). The combined organic layers were washed with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to obtain a crude compound. This crude compound was purified by column chromatography (100-200 mesh silica gel, eluent: 50-80% ethyl acetate in hexane) to give (2-allyl-1-(3-aminophenyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-6-yl)(1-methyl-1H-indazol-5-yl) tert-butyl carbamate (0.150 g, 0.234 mmol, 50.8% yield), a brown solid. LCMS m / z: 513.4 (M+H).

[0585] tert-Butyl-(2-allyl-1-(3-((1-methylpiperidin-4-yl)amino)phenyl)-3-oxo-2,3-dihydro- 1H-pyrazolo[3,4-d]pyrimidin-6-yl)(1-methyl-1H-indazol-5-yl)carbamate

[0586] Under an inert atmosphere, tert-butyl carbamate (0.15 g, 0.293 mmol) and 1-methylpiperidin-4-one (0.033 g, 0.293 mmol) in dichloroethane (5 mL) were mixed with AcOH (1 mL). The mixture was stirred at room temperature for 4 hours and then cooled to 0 °C. STAB (0.311 g, 0.293 mmol) was added and the mixture was stirred for 16 hours. The reaction progress was monitored by TLC and UPLC. After the reaction was complete, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (2 x 100 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude compound. This crude compound was purified by rapid column chromatography (100-200 mesh silica, eluent 0-20% MeOH and DCM) to give (2-allyl-1-(3-((1-methylpiperidin-4-yl)amino)phenyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-6-yl)(1-methyl-1H-indazole-5-yl)carbamate tert-butyl ester (140 mg, 0.204 mmol, 69.8% yield), a yellow solid.

[0587] tert-Butyl-(2-allyl-1-(3-(methyl(1-methylpiperidin-4-yl)amino)phenyl)-3-oxo-2,3-di Hydrogen-1H-pyrazolo[3,4-d]pyrimidin-6-yl)(1-methyl-1H-indazol-5-yl)carbamate

[0588] Under an inert atmosphere and at 0 °C, NaH (0.012 g, 0.533 mmol) was added to a stirred solution of (2-allyl-1-(3-((1-methylpiperidin-4-yl)amino)phenyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-6-yl)(1-methyl-1H-indazole-5-yl)carbamate (0.130 g, 0.213 mmol) in THF (2 mL), and the mixture was stirred for 30 min. Then, MeI (0.030 g, 0.213 mmol) was added at room temperature, and the mixture was stirred for 16 h. The reaction progress was monitored by TLC and UPLC. After the reaction was complete, the reaction mixture was diluted with water (100 mL) and extracted with DCM (2 x 100 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give crude compound (2-allyl-1-(3-(methyl(1-methylpiperidin-4-yl)amino)phenyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-6-yl)(1-methyl-1H-indazol-5-yl)carbamate tert-butyl ester (120 mg), a yellow solid. This crude compound was used in the next step without purification.

[0589] 2-Allyl-1-(3-(methyl(1-methylpiperidin-4-yl)amino)phenyl)-6-((1-methyl-1H-indazole-5-) (225)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one

[0590] A solution of 4 M HCl in 1,4-dioxane (0.3 mL, 1.200 mmol) was added to a stirred solution of (2-allyl-1-(3-(methyl(1-methylpiperidin-4-yl)amino)phenyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-6-yl)(1-methyl-1H-indazol-5-yl)carbamate (90 mg, 0.144 mmol) in DCM (5 mL), and the mixture was stirred at room temperature for 16 hours. The reaction progress was monitored by TLC and UPLC. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to obtain a crude compound, which was then purified by preparative HPLC (column: X-select CSH C18, mobile phase A: 0.1% aqueous formic acid, mobile phase B: acetonitrile, flow rate: 15.0 mL / min, Rt-10.8) to obtain 2-allyl-1-(3-(methyl(1-methylpiperidin-4-yl)amino)phenyl)-6-((1-methyl-1H-indazol-5-yl)amino)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (7.7 mg, 0.015 mmol, 10.09% yield), a grayish-white solid.

[0591] Example 65. Synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[m-(1-methyl-4-piperidinoxy)phenyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 220)

[0592] At 25 °C, 37% aqueous formaldehyde solution (172 mg, 2.114 mmol) was added to a stirred solution of 2-allyl-6-((1-methyl-1H-indazol-5-yl)amino)-1-(3-(piperidin-4-yloxy)phenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (210 mg, 0.423 mmol) in THF (5 mL), and the mixture was stirred for 10 min. STAB (269 mg, 1.269 mmol) was then added in portions. After complete addition of STAB, the reaction mixture was stirred at 25 °C for 16 h. The reaction progress was monitored by UPLC. After the reaction was complete, the reaction mixture was quenched with TFA, followed by ammonia in MeOH solution. The reaction mixture was diluted with water and extracted with 10% MeOH in DCM solution. The combined organic extracts were washed with an aqueous sodium bicarbonate solution, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain a crude compound. This crude compound was purified by preparative HPLC to give 2-allyl-6-((1-methyl-1H-indazol-5-yl)amino)-1-(3-((1-methylpiperidin-4-yl)oxy)phenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (21.22 mg, 0.040 mmol, 9.53% yield), a white solid.

[0593] Example 66. Synthesis of p-{2-ethyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazaindene-6-ylamino}benzyl nitrile (compound 310)

[0594] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 37.1 mg (44%).

[0595] Example 67. Synthesis of 2-allyl-6-(1-benzofuran-5-ylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 309)

[0596] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 154 mg TFA salt (84%), as a pale yellow powder.

[0597] Example 68. Synthesis of 2-allyl-6-(1-benzofuran-6-ylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 308)

[0598] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 114 mg TFA salt (62%), as a pale yellow powder.

[0599] Example 69. Synthesis of 2-allyl-6-(2H-1,3-benzodioxacyclopenten-5-ylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 307)

[0600] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 130 mg TFA salt (70%), as a pale yellow powder.

[0601] Example 70. Synthesis of m-{2-allyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazaindene-6-ylamino}benzyl nitrile (compound 305)

[0602] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 25 mg (27%).

[0603] Example 71. Synthesis of 2-allyl-6-(1,3-benzothiazol-6-ylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 303)

[0604] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 19 mg TFA salt (80%), as a pale yellow powder.

[0605] Example 72. Synthesis of p-{2-ethyl-3-oxo-1-[6-(4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-6-ylamino}benzyl nitrile (compound 302)

[0606] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridinylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, using p-bromobenzyl nitrile and tert-butyl 4-[(6-{6-amino-2-ethyl-3-oxo-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl}pyridin-2-yl)oxy]piperidin-1-carboxylic acid. Yield: 7.8 mg (9%).

[0607] Example 73. Synthesis of p-{2-allyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazaindene-6-ylamino}benzyl nitrile (compound 301)

[0608] This compound was prepared using p-bromobenzyl nitrile, in a manner similar to that described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridinylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0609] Example 74. p-{2-allyl-3-oxo-1-[6-(4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-6-ylamino}benzyl nitrile (compound 300)

[0610] 4-{6-[2-allyl-6-(p-cyanophenylamino)-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraaza-} Indene-1-yl]-2-pyridyloxy}-1-piperidinecarboxylate

[0611] tert-butyl-4-[6-(2-allyl-6-amino-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazaindene-1-yl)-2-pyridyloxy]-1-piperidinecarboxylate (200 mg, 0.428 mmol), p-bromobenzyl nitrile (155.8 mg, 0.856 mmol), and Cs₂CO₃ (418 mg, 3 equivalents) were mixed and stirred in dioxane (5 mL) under nitrogen for 15 min. Catalytic amounts of CuI (252.6 mg, 3.1 equivalents) and 1,2-bis(methylamino)ethane were added as ligands (41.5 mg, 0.471 mmol). The reaction mixture was heated to 90 °C and maintained for 48 h. The reaction progress was observed by LCMS. The reaction mixture was cooled to room temperature, and the crude residue was quenched with saturated ammonia solution, then extracted with EtOAc (3 × 10 mL) and washed with brine. The combined organic layers were dried (anhydrous Na₂SO₄) and the solvent was evaporated under reduced pressure. The crude product was dissolved in MeCN (5 ml) and crystallized over 1 hour by slow addition of water (5 ml). The light brown solid crude material was dissolved in DCM (5 ml), loaded onto a 10 g silica gel column, and purified by rapid chromatography (0-100%, EtOAc: PE) to give 4-{6-[2-allyl-6-(p-cyanophenylamino)-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazaindene-1-yl]-2-pyridyloxy}-1-piperidinecarboxylic acid tert-butyl ester [175 mg, 72% yield], as a yellow solid.

[0612] p-{2-allyl-3-oxo-1-[6-(4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5, 7-Tetraazainden-6-ylaminobenzyl nitrile, a trifluoroacetate (compound 300).

[0613] A solution of tert-butyl 4-{6-[2-allyl-6-(p-cyanophenylamino)-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazaindene-1-yl]-2-pyridyloxy}-1-piperidinecarboxylate (175 mg, 0.308 mmol) in DCM (3 mL) was treated with TFA (1.5 mL) for 2 hours. The solvent was removed under vacuum, and the crude product was dissolved in EtOAc, washed with saturated aqueous NaHCO3 solution and brine, dried (MgSO4), and concentrated. The resulting material was purified by reversed-phase chromatography (Gemini NX-C18, 21*150 mm, water (0.1% TFA) / acetonitrile, gradient over 12 minutes, 25 mL / min). The pure fractions were collected and concentrated to obtain p-{2-allyl-3-oxo-1-[6-(4-piperidinoxy)-2-pyridinyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-6-ylamino}benzyl nitrile, a trifluoroacetate, as a white solid [165 mg].

[0614] Example 75. 2-Allyl-6-(1,2-Phenylacetoxazol-6-ylamino)-1-[6-(1-Methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 299)

[0615] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 55 mg (42%), as a white powder.

[0616] Example 76. 2-Allyl-6-(1-Methyl-1H-indazol-5-ylamino)-1-[m-(4-piperidinyloxy)phenyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 223)

[0617] tert-butyl-4-(3-bromophenoxy)piperidine-1-carboxylate

[0618] Under an inert atmosphere, NaH (2.285 g, 57.1 mmol) was added to a stirred solution of tert-butyl 4-hydroxypiperidine-1-carboxylate (7.48 g, 37.1 mmol) in DMF (50 mL) at 0 °C, and the mixture was stirred at 50 °C for 1 h. The reaction mixture was cooled to room temperature, and a solution of 1-bromo-3-fluorobenzene (5 g, 28.6 mmol) dissolved in DMF (5 mL) was added. After the addition, the reaction mixture was stirred at 70 °C for 3 h. The reaction progress was monitored by LCMS and TLC. After the reaction was complete, the reaction mixture was quenched with ice-cold water and extracted with ethyl acetate (2 x 400 mL). The organic phases were combined and washed with brine (500 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to obtain a crude compound. This crude compound was purified by column chromatography (100-200 mesh silica, eluent: 10-20% ethyl acetate and hexane) to give tert-butyl 4-(3-bromophenoxy)piperidine-1-carboxylate (6.8 g, 14.89 mmol, 52.1% yield) as a yellow gel-like liquid. This separated product was used in the next step.

[0619] tert-butyl-4-(3-(2-allyl-6-(methylthio)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidine (Pyridine-1-yl)phenoxy)piperidine-1-carboxylate

[0620] Under an inert atmosphere, a stirred solution of 4-(3-bromophenoxy)piperidin-1-carboxylic acid tert-butyl ester (481 mg, 1.350 mmol), 2-allyl-6-(methylthio)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (250 mg, 1.125 mmol), K₂CO₃ (466 mg, 3.37 mmol), and N,N′-dimethylethylenediamine (99 mg, 1.125 mmol) in dioxane (10 mL) was degassed for 20 min at room temperature. CuI (214 mg, 1.125 mmol) was then added, and the mixture was degassed again for 5 min. The reaction mixture was stirred at 100 °C for 16 h. The reaction progress was monitored by TLC and LCMS. After the reaction was complete, the reaction mixture was diluted with DCM and filtered through a diatomaceous earth mat. The collected organic fractions were concentrated under reduced pressure to obtain a crude compound, which was then purified by rapid column chromatography (SiO2 / 230-400 mesh; 20-50% ethyl acetate-petroleum ether) to give tert-butyl-4-(3-(2-allyl-6-(methylthio)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylate (80 mg, 0.154 mmol, 13.72% yield), as a grayish-white solid.

[0621] tert-butyl-4-(3-(2-allyl-6-(methanesulfonyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]) Pyrimidin-1-yl)phenoxy)piperidine-1-carboxylate

[0622] At room temperature, m-CPBA (55.5 mg, 0.322 mmol) was added to a stirred solution of 4-(3-(2-allyl-6-(methylthio)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylic acid tert-butyl ester (80 mg, 0.161 mmol) in DCM (5 mL), and the mixture was stirred for 2 hours. The reaction progress was monitored by TLC and LCMS. After the reaction was complete, it was quenched with sodium bicarbonate solution and then extracted with 10% MeOH in DCM solution (2 x 50 mL). The combined organic layers were dried over sodium sulfate and evaporated under reduced pressure to give tert-butyl 4-(3-(2-allyl-6-(methanesulfonyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylate (85 mg, 0.053 mmol, 32.9% yield). This compound was used in the next step without further purification.

[0623] tert-butyl-4-(3-(2-allyl-6-((1-methyl-1H-indazol-5-yl)amino)-3-oxo-2,3-dihydro- 1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylate

[0624] At room temperature, 1-methyl-1H-indazole-5-amine (18.06 mg, 0.123 mmol) was added to a stirred solution of 4-(3-(2-allyl-6-(methanesulfonyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylic acid tert-butyl ester (65 mg, 0.123 mmol) in AcOH (3 mL). The reaction mixture was stirred for 16 hours. The reaction progress was monitored by TLC and LCMS. After the reaction was complete, the reactants were quenched with aqueous sodium bicarbonate solution and extracted with 10% MeOH in DCM solution (2 x 100 mL). The combined organic layers were dried over Na2SO4 and then concentrated under reduced pressure to obtain a crude compound, which was purified by column chromatography (SiO2 / 230-400 mesh; 0-20% MeOH-DCM) to give tert-butyl 4-(3-(2-allyl-6-((1-methyl-1H-indazol-5-yl)amino)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylic acid (30 mg). This separated product was used in the next step.

[0625] 2-Allyl-6-((1-Methyl-1H-indazol-5-yl)amino)-1-(3-(piperidin-4-yloxy)phenyl)-1,2- Dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 223)

[0626] Under an inert atmosphere, at 0 °C, a solution of 4 M HCl in dioxane (0.2 mL) was added to a stirred solution of 30 mg (0.050 mmol) of 4-(3-(2-allyl-6-((1-methyl-1H-indazol-5-yl)amino)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylic acid in dioxane (1 mL), and the mixture was stirred for 16 hours. The reaction progress was monitored by TLC and LCMS. After the reaction was complete, the solvent was evaporated under reduced pressure and washed with n-hexane (2 x 5 mL). The solution was then purified by preparative HPLC (column: X-select CSH C18, mobile phase A: water (containing 0.1% AA), mobile phase B: acetonitrile, flow rate -15.0 mL / min, Rt -12.8) to give 2-allyl-6-((1-methyl-1H-indazol-5-yl)amino)-1-(3-(piperidin-4-yloxy)phenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (4 mg, 7.97 µmol, 15.86% yield) as a brown solid.

[0627] Example 77. 2-Allyl-6-(1-Methyl-1H-indazol-5-ylamino)-1-[m-(1-methyl-4-piperidinylamino)phenyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 226)

[0628] 2-Allyl-6-(methylthio)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one

[0629] At room temperature, 3-nitrophenylboronic acid (1.352 g, 8.10 mmol), sodium carbonate (1.707 g, 16.20 mmol), and copper(II) acetate (0.490 g, 2.70 mmol) were added to a stirred solution of 2-allyl-6-(methylthio)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (1,1.2 g, 5.40 mmol) in dichloroethane (15 mL), followed by the addition of pyridine (0.169 g, 1.080 mmol). The temperature was raised to 70 °C and the mixture was stirred for 16 hours. The reaction progress was monitored by TLC and LCMS. After the reaction was complete, the reaction mixture was filtered through diatomaceous earth and extracted with 10% MeOH in DCM solution (2 x 100 mL). The combined organic layers were dried over anhydrous Na2SO4 and then evaporated under reduced pressure to obtain a crude compound. This crude compound was purified by column chromatography (100-200 mesh silica, eluent 70-100% ethyl acetate and hexane) to give 2-allyl-6-(methylthio)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (700 mg, 1.794 mmol, 33.2% yield) as a white solid.

[0630] 2-Allyl-6-(Methylsulfonyl)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidine- 3-keto

[0631] Under an inert atmosphere, at room temperature, 2-allyl-6-(methylthio)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (50 mg, 0.146 mmol) was added to a stirred solution of 2-allyl-6-(methylthio)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (10 ml) in DCM. mCPBA (53.7 mg, 0.218 mmol) was added, and stirring continued for 2 hours. The reaction progress was monitored by TLC and UPLC. After the reaction was complete, the reaction mixture was quenched with 10% sodium bicarbonate aqueous solution (10 mL) and extracted with 10% MeOH in DCM solution (2 x 50 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude compound 2-allyl-6-(methanesulfonyl)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (30 mg, 0.024 mmol, 16.47% yield) as a grayish-white solid. This crude compound was used for the next step without any further purification.

[0632] 2-Allyl-6-((1-Methyl-1H-indazol-5-yl)amino)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyridine Azo[3,4-d]pyrimidin-3-one

[0633] At room temperature, 1-methyl-1H-indazole-5-amine (0.3 g, 2.039 mmol) was added to a stirred solution of 2-allyl-6-(methylthio)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (0.7 g, 2.039 mmol) in AcOH (5 mL) and stirred for 16 hours. The reaction progress was monitored by TLC and UPLC. After the reaction was complete, the solvent was evaporated under reduced pressure, and the residue was quenched with 10% sodium bicarbonate aqueous solution (100 mL). The mixture was extracted with ethyl acetate (2 x 300 mL). The combined organic extracts were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude compound. This crude compound was purified by column chromatography (100-200 mesh silica gel, eluent 10-20% ethyl acetate and hexane) to give 2-allyl-6-((1-methyl-1H-indazol-5-yl)amino)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (600 mg, 1.221 mmol, 66.51% yield) as a yellow solid.

[0634] 2-Allyl-1-(3-aminophenyl)-6-((1-methyl-1H-indazol-5-yl)amino)-1,2-dihydro-3H-pyridine Azo[3,4-d]pyrimidin-3-one

[0635] Iron (63.1 mg, 1.130 mmol) and NH4Cl (60.4 mg, 1.130 mmol) were added to a stirred solution of 2-allyl-6-((1-methyl-1H-indazol-5-yl)amino)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (50 mg, 0.113 mmol) in EtOH (3.0 mL) and water (10 mL). The temperature was raised to 60 °C and the mixture was stirred for 2 hours. The reaction progress was monitored by TLC and UPLC. After the reaction was complete, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (2 x 300 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude compound. This crude compound was purified by rapid column chromatography (100-200 mesh silica, eluent of 50-80% ethyl acetate and hexane) to give 2-allyl-1-(3-aminophenyl)-6-((1-methyl-1H-indazol-5-yl)amino)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (200 mg, 0.023 mmol, 20.38% yield), a white solid.

[0636] 2-Allyl-6-((1-Methyl-1H-indazol-5-yl)amino)-1-(3-((1-methylpiperidin-4-yl)amino)benzene (226)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one:

[0637] AcOH (1 mL) was added to a stirred solution of 2-allyl-1-(3-aminophenyl)-6-((1-methyl-1H-indazol-5-yl)amino)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (0.2 g, 0.485 mmol) and 1-methylpiperidin-4-one (55 mg, 0.485 mmol) at room temperature, and the mixture was stirred for 2 hours. The mixture was then cooled to 0 °C, STAB (0.617 g, 2.91 mmol) was added, and the mixture was stirred at room temperature for 16 hours. The reaction progress was monitored by TLC and LCMS. After the reaction was complete, the reaction mixture was quenched with ice-cold water and extracted with DCM (2 x 100 mL). The combined organic layers were dried over anhydrous Na2SO4 and evaporated under reduced pressure to obtain a crude compound. This crude compound was purified by preparative HPLC (column: X-select CSH C18, mobile phase A: 0.1% aqueous formic acid, mobile phase B: acetonitrile, flow rate: 15.0 mL / min, Rt-10.8) to obtain 2-allyl-6-((1-methyl-1H-indazol-5-yl)amino)-1-(3-((1-methylpiperidin-4-yl)amino)phenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (14 mg, 0.026 mmol, 5.44% yield), a grayish-white solid.

[0638] Example 78. 2-Ethyl-1-[6-(4-piperidinoxy)-2-pyridyl]-6-m-toluidine-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 298)

[0639] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 55 mg (42%), as a white powder. Yield: 92.5 mg, 80%, as a grayish-white solid.

[0640] Example 79. 2-Ethyl-6-(p-fluorophenylamino)-1-[6-(4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 297)

[0641] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 55 mg (42%), as a white powder. Yield: 50.0 mg, 43%, as a grayish-white solid.

[0642] Example 80. 2-Allyl-6-(1-methyl-4-pyrazolylamino)-1-[m-(4-piperidinyloxy)phenyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 224)

[0643] tert-butyl-4-(3-(2-allyl-6-((1-methyl-1H-pyrazol-4-yl)amino)-3-oxo-2,3-dihydro- 1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylate

[0644] At 25 °C, 1-methyl-1H-pyrazol-4-amine (14.67 mg, 0.151 mmol) was added to a stirred solution of 4-(3-(2-allyl-6-(methanesulfonyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylic acid tert-butyl ester (80 mg, 0.151 mmol) in acetic acid (3 mL). The reaction mixture was stirred at 25 °C for 16 h. The reaction progress was monitored by TLC and LCMS. After the reaction was complete, the reaction mixture was concentrated under reduced pressure, diluted with 10% aqueous sodium bicarbonate solution, and extracted with 10% MeOH in DCM solution. The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude compound. This crude compound was purified by column chromatography (silica gel 100-200 mesh, eluent 5-10% MeOH in DCM solution) to give tert-butyl 4-(3-(2-allyl-6-((1-methyl-1H-pyrazol-4-yl)amino)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylic acid (60 mg, 0.082 mmol, 54.5% yield), as a grayish-white solid.

[0645] 2-Allyl-6-((1-methyl-1H-pyrazol-4-yl)amino)-1-(3-(piperidin-4-yloxy)phenyl)-1,2- Dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 224)

[0646] Under an inert atmosphere and at 0 °C, a solution of 4 M HCl (0.329 mL, 1.317 mmol) in dioxane was added to a stirred solution of 60 mg (0.110 mmol) of 4-(3-(2-allyl-6-((1-methyl-1H-pyrazol-4-yl)amino)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylic acid in 1,4-dioxane (5 mL). The reaction was stirred at room temperature for 8 hours. The progress of the reaction was monitored by TLC and LCMS. After the reaction was completed, the crude product was concentrated under reduced pressure and then passed through preparative HPLC (column: X-select CSH C18, mobile phase A: water (containing 0.1% FA), mobile phase B: acetonitrile (containing 0.1% FA), flow rate: 15.0 mL / min, Rt: 12.8) to obtain 2-allyl-6-((1-methyl-1H-pyrazol-4-yl)amino)-1-(3-(piperidin-4-yloxy)phenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (8 mg, 0.018 mmol, 16% yield), a brown solid.

[0647] Example 81. 2-Allyl-1-[6-(4-piperidinoxy)-2-pyridyl]-6-[p-(2,2,2-trifluoroethoxy)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 227)

[0648] tert-butyl-4-(6-{2-allyl-3-oxo-6-[p-(2,2,2-trifluoroethoxy)phenylamino]-1,2-di Hydrogen-3H-1,2,5,7-tetraazainden-1-yl}-2-pyridyloxy)-1-piperidinecarboxylate

[0649] Under nitrogen atmosphere, at room temperature, 166 mg (assumed 0.722 mmol) of mCPBA (<77% pure) in DCM (0.5 mL) was added to a stirred solution of 4-{6-[2-allyl-6-(methylthio)-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazaindene-1-yl]-2-pyridyloxy}-1-piperidinecarboxylate tert-butyl ester (300 mg, 0.602 mmol) in DCM (2.5 mL). After 15 minutes, the DCM was concentrated under vacuum, and the crude product was dissolved in MeCN (3 mL). Then, p-(2,2,2-trifluoroethoxy)aniline (115 mg, 0.602 mmol) was added, and the reaction mixture was stirred in a sealed vial at 60 °C. After 18 hours, the reaction mixture was cooled to room temperature, and mCPBA quenched with 1 M NaOH (5 mL) was added dropwise. The aqueous phase was extracted with EtOAc (3 x 20 mL) and washed with brine (20 mL). The combined organic layers were dried (MgSO4) and concentrated under reduced pressure to give a yellow powder. The crude material was dissolved in DCM (5 mL), loaded onto a 10 g silica gel column, and purified by rapid chromatography (0-100%, EtOAc: PE) to give tert-butyl 4-(6-{2-allyl-3-oxo-6-[p-(2,2,2-trifluoroethoxy)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazainden-1-yl}-2-pyridyloxy)-1-piperidinecarboxylate [210 mg, 94%], as a pale yellow solid.

[0650] 2-Allyl-6-[p-(2-fluoroethoxy)phenylamino]-1-[6-(4-piperidinyloxy)-2-pyridyl]-1, 2-Dihydro-3H-1,2,5,7-Tetraazainden-3-one trifluoroacetate (compound 227)

[0651] A solution of tert-butyl 4-(6-{2-allyl-3-oxo-6-[p-(2,2,2-trifluoroethoxy)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-1-yl}-2-pyridyloxy)-1-piperidinecarboxylate (210 mg, 0.360 mmol) in DCM (5 mL) was treated with TFA (2.5 mL) for 2 hours. The solvent was removed under vacuum, and the crude product was dissolved in EtOAc, washed with saturated aqueous NaHCO3 solution and brine, dried (MgSO4), and concentrated. The resulting material was purified by reversed-phase chromatography (Gemini NX-C18, 21*150 mm, water (0.1% TFA) / acetonitrile, gradient over 12 minutes, 25 mL / min). The purified fractions were collected and concentrated to give 2-allyl-6-[p-(2-fluoroethoxy)phenylamino]-1-[6-(4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, a trifluoroacetate, as a white solid. [1.4 mg].

[0652] Example 82. 5-{2-allyl-3-oxo-1-[6-(4-piperidinyloxy)-2-pyridinyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-6-ylamino}-2-toluenenitrile (Compound 293)

[0653] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridinylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 27 mg, 47%.

[0654] Example 83. 2-Ethyl-6-(p-fluorophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 290)

[0655] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 55 mg (42%), as a white powder. Yield: 156 mg, 43%, as a yellow solid.

[0656] Example 84. 2-Ethyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-m-toluidine-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 289)

[0657] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 55 mg (42%), as a white powder. Yield: 82 mg, 53%, as a grayish-white solid.

[0658] Example 85. 2-Allyl-1-(6-(methyl(1-methylpiperidin-4-yl)amino)pyridin-2-yl)-6-((1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl)amino)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 201)

[0659] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[m-(1-methyl-4-piperidinylamino)phenyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 28 mg (25.2%).

[0660] Example 86. 2-Allyl-6-(1-Isobutyl-4-pyrazolylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 200)

[0661] 4-{6-[6-(1-isobutylpyrazole-4-ylamino)-3-oxo-2-(prop-2-enyl)-1,2-dihydro-3H-1,2, 5,7-Tetraazaindene-1-yl]pyridin-2-yloxy}piperidine-1-carboxylic acid tert-butyl ester

[0662] Under nitrogen atmosphere, at room temperature, 108 mg (assumed 0.469 mmol) of mCPBA (<77% pure) in DCM (0.5 mL) was added to a stirred solution of 4-{6-[2-allyl-6-(methylthio)-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazaindene-1-yl]-2-pyridyloxy}-1-piperidinecarboxylic acid tert-butyl ester (123 mg, 0.247 mmol) in DCM (5 mL). After 15 minutes, the DCM was concentrated under vacuum, and the crude product was dissolved in MeCN (5 mL). Then, 1-isobutyl-4-pyrazolylamine (54.4 mg, 0.391 mmol) was added. The reaction mixture was stirred in a sealed vial at 60 °C. 22 After hours, the reaction mixture was cooled to room temperature, and mCPBA quenched with 1M NaOH (5 mL) was added dropwise. The aqueous phase was extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine, dried (MgSO4), and concentrated under reduced pressure to give a crude product as a yellow powder. The crude material was dissolved in DCM (5 mL), loaded onto a 10 g silica gel column, and purified by rapid chromatography (0-100%, EtOAc:PE) to give 4-{6-[6-(1-isobutylpyrazole-4-ylamino)-3-oxo-2-(prop-2-enyl)-1,2-dihydro-3-ylamino] H -1,2,5,7-Tetraazaindene-1-yl]pyridin-2-yloxy}piperidine-1-carboxylic acid Uncle Butyl ester, a pale yellow solid (100 mg, 84%).

[0663] 2-Allyl-6-(1-Isobutyl-4-pyrazolylamino)-1-[6-(4-piperidinyloxy)-2-pyridyl]-1,2- Dihydro-3H-1,2,5,7-tetraazainden-3-one

[0664] 4-{6-[6-(1-isobutylpyrazole-4-ylamino)-3-oxo-2-(prop-2-enyl)-1,2-dihydro-3- H A solution of tert-butyl [-1,2,5,7-tetraazaindene-1-yl]pyridin-2-yloxy}piperidine-1-carboxylate (100 mg, 0.205 mmol) in DCM (5 mL) was treated with TFA (2 mL) for 2 h. The solvent was removed under vacuum, and the crude product was dissolved in EtOAc, washed with saturated aqueous solution of NaHCO3 and brine, dried (MgSO4), and concentrated. The resulting material was purified by HPLC (Phenomenex Gemini 5 µm NX-C18 110 Å 150x21, 2 mm, buffer 0.2% NH4OH, water (0.2% NH4OH) / acetonitrile, gradient within 9 min, 30 mL / min). The pure fractions were collected and concentrated to give 2-allyl-6-(1-isobutyl-4-pyrazolylamino)-1-[6-(4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, a white solid (100 mg).

[0665] 2-Allyl-6-(1-Isobutyl-4-pyrazolylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridine] [1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 200)]

[0666] 2-Allyl-6-(1-isobutyl-4-pyrazolylamino)-1-[6-(4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (100 mg, 0.205 mmol) was dissolved in anhydrous THF (5 mL). Formaldehyde (approximately 36% pure, 30.4 µL, 0.408 mmol) and STAB (86.5 mg, 0.408 mmol) were added sequentially. The reaction mixture was stirred at room temperature while being monitored by LCMS. After 2 hours, the reaction was quenched dropwise with saturated sodium bicarbonate aqueous solution (5 mL). The aqueous phase was extracted with EtOAc (3 x 20 mL). The combined organic layers were dried (MgSO4) and concentrated under reduced pressure to give a crude material as a yellow powder. The crude product was dissolved in MeOH (5 ml) and purified by HPLC (Phenomenex Gemini 5 µmNX-C18 110Å 150x21, 2 mm, buffer 0.2% NH4OH, water (0.2% NH4OH) / acetonitrile, gradient over 6 min, 30 ml / min). The purified fractions were collected and concentrated to give 2-allyl-6-(1-isobutyl-4-pyrazolylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (60 mg, 60%).

[0667] Example 87. 2-Allyl-6-(1-Isopropyl-4-pyrazolylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 198)

[0668] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 60 mg (24%).

[0669] Example 88. 2-Allyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-(6-methyl-3-pyridylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 197)

[0670] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 29 mg TFA salt (83%), as a pale yellow powder.

[0671] Example 89. 2-Allyl-6-(2H-1,3-benzodioxacyclopenten-5-ylamino)-1-[6-(1-methyl-4-piperidinylamino)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 196)

[0672] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[m-(1-methyl-4-piperidinylamino)phenyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. The purified material was converted to a free base by solid-phase extraction (1 g SCX-2, MeOH, NH3 / MeOH 1.4 M). Yield: 41 mg of free base (33%), as a brown solid.

[0673] Example 90. 2-Allyl-6-(1-benzofuran-6-ylamino)-1-[6-(1-methyl-4-piperidinylamino)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 195)

[0674] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[m-(1-methyl-4-piperidinylamino)phenyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 110 mg TFA salt (74%), as a yellow solid.

[0675] Example 91. 2-Allyl-1-{6-(9-methyl-9-azabicyclo[3.3.1]non-3-yloxy)-2-pyridyl}-6-(1-methyl-4-pyrazolylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 194)

[0676] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 40 mg (31.7%).

[0677] Example 92. 1-{6-[(R)-3-piperidinoxy]-2-pyridyl}-2-allyl-6-(p-chlorophenylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 192)

[0678] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one.

[0679] Example 93. 2-Allyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-m-toluidine-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 191)

[0680] 4-{6-[6-(3-methylphenylamino)-3-oxo-2-(prop-2-enyl)-1,2-dihydro-3H-1,2,5,7-tetra- [Za-indene-1-yl]pyridin-2-yloxy}piperidine-1-carboxylic acid tert-butyl ester

[0681] Under nitrogen atmosphere, at room temperature, mCPBA (<77% pure) (167.2 mg, assumed 0.727 mmol) in DCM (0.5 mL) was added to a stirred solution of 4-{6-[2-allyl-6-(methylthio)-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazaindene-1-yl]-2-pyridyloxy}-1-piperidinecarboxylic acid tert-butyl ester (300 mg, 0.602 mmol) in DCM (5 mL). After 15 minutes, the DCM was concentrated under vacuum, and the crude product was dissolved in MeCN (5 mL), followed by the addition of m-toluidine (96.7 mg, 0.903 mmol). The reaction mixture was stirred in a sealed vial at 60 °C. 22 After hours, the reaction mixture was cooled to room temperature, and mCPBA quenched with 1M NaOH (5 mL) was added dropwise. The aqueous phase was extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (20 mL), dried (MgSO4), and concentrated under reduced pressure to give a crude product as a yellow powder. The crude material was dissolved in DCM (5 mL), loaded onto a 10 g silica gel column, and purified by rapid chromatography (0-100%, EtOAc:PE) to give 4-{6-[6-(3-methylphenylamino)-3-oxo-2-(prop-2-enyl)-1,2-dihydro-3- H -1,2,5,7-Tetraazaindene-1-yl]pyridin-2-yloxy}piperidine-1-carboxylic acid Uncle Butyl ester [300 mg, 87.2%] is a light yellow solid.

[0682] 2-Allyl-1-[6-(4-piperidinyloxy)-2-pyridyl]-6-m-tolyl-1,2-dihydro-3H-1,2, 5,7-Tetraazaindene-3-one trifluoroacetate

[0683] 4-{6-[6-(3-methylphenylamino)-3-oxo-2-(prop-2-enyl)-1,2-dihydro-3- HA solution of tert-butyl [-1,2,5,7-tetraazaindene-1-yl]pyridin-2-yloxy}piperidine-1-carboxylate (300 mg, 0.525 mmol) in DCM (5 mL) was treated with TFA (2 mL) for 2 h. The solvent was removed under vacuum, and the crude product was dissolved in EtOAc, washed with saturated aqueous solution of NaHCO3 and brine, dried (MgSO4), and concentrated. The resulting material was purified by HPLC (Phenomenex Gemini 5µm NX-C18 110Å 150x21, 2 mm, buffer 0.2% NH4OH, water (0.2% NH4OH) / acetonitrile, gradient within 9 min, 30 mL / min). The pure fractions were collected and concentrated to give 2-allyl-1-[6-(4-piperidinyloxy)-2-pyridyl]-6-m-toluidine-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, a trifluoroacetate, as a white solid [120 mg].

[0684] 2-Allyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-m-toluidine-1,2-dihydro- 3H-1,2,5,7-Tetraazainden-3-one (Compound 191)

[0685] 2-Allyl-1-[6-(4-piperidinoxy)-2-pyridyl]-6-m-toluidine-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (120 mg, 0.21 mmol) was dissolved in anhydrous THF (5 mL). Formaldehyde (approximately 36% pure, 31 µL, 0.21 mmol) and STAB (89 mg, 0.21 mmol) were added sequentially. The reaction mixture was stirred at room temperature and monitored by LCMS. After 2 hours, the reaction was quenched dropwise with saturated sodium bicarbonate aqueous solution (5 mL). The aqueous phase was extracted with EtOAc (3 x 20 mL). The combined organic layers were dried (MgSO4) and concentrated under reduced pressure to give a crude material as a yellow powder.

[0686] The crude product was dissolved in MeOH (5 ml) and purified by HPLC (Phenomenex Gemini 5 µm NX-C18110Å 150x21, 2 mm, buffer 0.2% NH4OH, water (0.2% NH4OH) / acetonitrile, gradient over 6 min, 30 ml / min). The purified fractions were collected and concentrated to give 2-allyl-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridinyl]-6-m-toluidine-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (40 mg, 33%) as a white solid [100 mg, 33% yield].

[0687] Example 94. 2-Allyl-1-[6-(1-methyl-4-piperidinylamino)-2-pyridyl]-6-m-toluidine-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 188)

[0688] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[m-(1-methyl-4-piperidinylamino)phenyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 144 mg, 56%.

[0689] Example 95. 2-Allyl-1-[6-(1-methyl-4-piperidinylamino)-2-pyridyl]-6-[4-(2,2,2-trifluoroethoxy)-3-toluidine]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 181)

[0690] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[m-(1-methyl-4-piperidinylamino)phenyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 105 mg, 68%.

[0691] Example 96. 2-Allyl-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridinyl]-6-(1-propyl-4-pyrazolylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 180)

[0692] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 75 mg, 70%.

[0693] Example 97. 2-Allyl-1-[6-(4-piperidinylamino)-2-pyridyl]-6-[4-(2,2,2-trifluoroethoxy)-3-toluidine]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 178)

[0694] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[m-(1-methyl-4-piperidinylamino)phenyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 220 mg, 81%.

[0695] Example 98. 1-{6-[N-methyl(1-methyl-4-piperidinyl)amino]-2-pyridinyl}-2-allyl-6-(1-propyl-4-pyrazolylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 177)

[0696] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[m-(1-methyl-4-piperidinylamino)phenyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 8 mg, 10%.

[0697] Example 99. 1-{6-[N-(S)-3-piperidinyl-N-methylamino]-2-pyridinyl}-2-allyl-6-(p-chlorophenylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 173)

[0698] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[m-(1-methyl-4-piperidinylamino)phenyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, starting with tert-butyl (S)-3-amino-1-piperidincarboxylate. Yield: 7.0 mg TFA salt (31%).

[0699] Example 100. 1-{6-[N-(R)-3-piperidinyl-N-methylamino]-2-pyridinyl}-2-allyl-6-(p-chlorophenylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 171)

[0700] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[m-(1-methyl-4-piperidinylamino)phenyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, starting with (R)-3-amino-1-piperidinic acid tert-butyl ester. Yield: 6.6 mg TFA salt (25%).

[0701] Example 101. 1-{6-[(R)-1-methyl-3-piperidinylamino]-2-pyridyl}-2-allyl-6-(p-chlorophenylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 168)

[0702] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, starting with 1-{6-[(R)-3-piperidinylamino]-2-pyridyl}-2-allyl-6-(p-chlorophenylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 93 mg, 76%.

[0703] Example 102. 1-{6-[(S)-1-methyl-3-piperidinylamino]-2-pyridyl}-2-allyl-6-(p-chlorophenylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 167)

[0704] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, starting with 1-{6-[(S)-3-piperidinylamino]-2-pyridyl}-2-allyl-6-(p-chlorophenylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 91 mg, 34%.

[0705] Example 103. 1-{6-[(R)-3-piperidinylamino]-2-pyridyl}-2-allyl-6-(p-chlorophenylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 166)

[0706] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, starting with tert-butyl (R)-3-amino-1-piperidincarboxylate. Yield: 150 mg, 63%.

[0707] Example 104. 1-{6-[(S)-3-piperidinylamino]-2-pyridyl}-2-allyl-6-(p-chlorophenylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 165)

[0708] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, starting with tert-butyl (S)-3-amino-1-piperidincarboxylate. Yield: 310 mg, 39%.

[0709] Example 105. 1-(6-{[(S)-1-methyl-3-piperidinyl]-N-methylamino}-2-pyridinyl)-2-allyl-6-(p-chlorophenylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 164)

[0710] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, starting with tert-butyl (S)-3-amino-1-piperidincarboxylate. Yield: 40 mg TFA salt (43%), as a pale yellow solid.

[0711] Example 106. (R)-2-allyl-6-((4-chlorophenyl)amino)-1-(6-(methyl(1-methylpiperidin-3-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 163)

[0712] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, starting with tert-butyl (R)-3-amino-1-piperidincarboxylate. Yield: 38 mg TFA salt (35%), as a pale yellow solid.

[0713] Example 107. 2-Allyl-6-(3-fluoro-5-toluidine)-1-[6-(1-methyl-4-piperidinylamino)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 161)

[0714] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one. Yield: 54 mg, 62%.

[0715] Example 108. 2-Allyl-6-(3-fluoro-5-toluidine)-1-[6-(4-piperidinylamino)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 160)

[0716] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one. Yield: 78 mg, 63%.

[0717] Example 109. 2-Allyl-1-[6-(1-methyl-4-piperidinylamino)-2-pyridyl]-6-(3-phenyl-5-isothiazolylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 158)

[0718] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one. Yield: 1.8 mg, 37%.

[0719] Example 110. 6-(4-fluoro-3-toluidine)-2-methyl-1-[6-(4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 154)

[0720] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using 4-({6-[2-methyl-6-(methylthioalkyl)-3-oxo-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]pyridin-2-yl}oxy)piperidin-1-carboxylic acid tert-butyl ester (81 mg) and 4-fluoro-3-methylaniline (1 equivalent) to give a crude free base intermediate (77 mg), of which 20% was purified by preparative HPLC. Yield: 9.3 mg TFA salt (60%).

[0721] Example 111. 6-(p-chlorophenylamino)-2-methyl-1-[6-(4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 153)

[0722] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)oxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using 4-({6-[2-methyl-6-(methylthioalkyl)-3-oxo-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]pyridin-2-yl}oxy)piperidin-1-carboxylic acid tert-butyl ester (81 mg) and 4-chloroaniline (1 equivalent) to give a crude free base intermediate (78 mg), of which 20% was purified by preparative HPLC. Yield: 8.7 mg TFA salt (56%).

[0723] Example 112. 6-(4-fluoro-3-toluidine)-2-methyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 152)

[0724] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)oxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using 6-[(4-fluoro-3-methylphenyl)amino]-2-methyl-1-[6-(piperidin-4-yloxy)pyridin-2-yl]-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (62 mg). Yield: 57 mg TFA salt (71%), as a powder.

[0725] Example 113. 2-Allyl-6-(3,4-dichlorophenylamino)-1-[6-(1-methyl-4-piperidinylamino)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 151)

[0726] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one. Yield: 33 mg, 84%.

[0727] Example 114. 6-(p-chlorophenylamino)-2-methyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 150)

[0728] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 45 mg TFA salt (57%), as a powder.

[0729] Example 115. 1-{6-[(S)-1-methyl-3-pyrrolidinoxy]-2-pyridyl}-2-allyl-6-(p-chlorophenylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 148)

[0730] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, with tert-butyl (3S)-3-hydroxypyrrolidine-1-carboxylate. Yield: 94 mg TFA salt (82%), as a pale yellow powder.

[0731] Example 116. 2-Allyl-6-(3,4-dichlorophenylamino)-1-[6-(4-piperidinylamino)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 146)

[0732] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using 3,4-dichloroaniline and 4-({6-[6-(methylthioalkyl)-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]pyridin-2-yl}amino)piperidin-1-carboxylic acid tert-butyl ester. Yield: 46 mg, 24%.

[0733] Example 117. 1-{6-[(S)-3-pyrrolidinoxy]-2-pyridyl}-2-allyl-6-(p-chlorophenylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 145)

[0734] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, with tert-butyl (3S)-3-hydroxypyrrolidine-1-carboxylate. Yield: 177 mg TFA salt (62%), as a yellow powder.

[0735] Example 118. 2-Allyl-6-(p-chlorophenylamino)-1-[6-(4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 141)

[0736] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, using tert-butyl 4-({6-[6-(methylthioalkyl)-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyridin-1-yl]pyridin-2-yl}oxy)piperidin-1-carboxylic acid (99 mg) and 4-chloroaniline (1 equivalent). After deprotection, the reaction mixture was concentrated to a yellow oil (175 mg, crude), half of which was purified by preparative HPLC and then converted to a free base by solid-phase extraction (1 g SCX-2, MeOH, 1.4 M MeOH / NH3). Yield: 15 mg free base (31%), in powder form.

[0737] Example 119. 2-Allyl-6-(p-chlorophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 142)

[0738] 2-Allyl-1-(6-Fluoropyridin-2-yl)-6-(methylthio)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidine- 3-keto

[0739] At 25 °C, a stirred solution of 2-allyl-6-(methylthio)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (1 g, 4.50 mmol), 2-bromo-6-fluoropyridine (0.871 g, 4.95 mmol), trans-N,N-dimethylethylenediamine (0.397 g, 4.50 mmol), and K₂CO₃ (1.865 g, 13.50 mmol) in DMSO (4 mL) was degassed for 5 min, followed by the addition of copper(I) iodide (0.857 g, 4.50 mmol), and the resulting mixture was stirred at 100 °C for 2 h. The reaction progress was monitored by TLC. After the reaction was complete, the reaction mixture was filtered through a diatomaceous earth mat and washed with ethyl acetate (250 mL). The filtrate was concentrated under reduced pressure. The crude material was purified by rapid chromatography (SiO2 / 100-200 mesh; 20-30% ethyl acetate-hexane) to obtain 2-allyl-1-(6-fluoropyridin-2-yl)-6-(methylthio)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (700 mg, 2.089 mmol, 46.4% yield), a light brown solid.

[0740] 2-Allyl-1-(6-Fluoropyridin-2-yl)-6-(Methylsulfonyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidine 3-Pyridoxine

[0741] At 0 °C, m-CPBA (1.554 g, 6.30 mmol) was added to a stirred solution of 2-allyl-1-(6-fluoropyridin-2-yl)-6-(methylthio)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (1 g, 3.15 mmol) in dichloromethane (30 mL), and the mixture was stirred at 25 °C for 2 h. The reaction progress was monitored by TLC. After the reaction was complete, the reaction mixture was diluted with DCM (300 mL) and washed with sodium bicarbonate solution (2 x 150 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give crude 2-allyl-1-(6-fluoropyridin-2-yl)-6-(methanesulfonyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (1 g, 2.63 mmol, 84% yield), a pale yellow solid.

[0742] 2-Allyl-6-((4-chlorophenyl)amino)-1-(6-fluoropyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4- d]Pyrimidine-3-one

[0743] Under a nitrogen atmosphere, 4-chloroaniline (0.438 g, 3.44 mmol) was added to a stirred solution of 2-allyl-1-(6-fluoropyridin-2-yl)-6-(methanesulfonyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (1 g, 2.86 mmol) in acetic acid (10 mL) at room temperature. The reaction mixture was stirred at room temperature for 16 hours. The reaction progress was monitored by TLC. After the reaction was complete, the reaction mixture was concentrated under reduced pressure to give a residue. The residue was alkalized with sodium bicarbonate and extracted with ethyl acetate (10 x 2 mL). The combined organic layers were dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give a crude product. The crude material was purified by rapid chromatography (SiO2 / 100-200 mesh; 7-8% methanol-DCM) to obtain 2-allyl-6-((4-chlorophenyl)amino)-1-(6-fluoropyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (800 mg, 1.774 mmol, 62.0% yield), a light brown solid.

[0744] 2-Allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)oxy)pyridin-2-yl)-1, 2-Dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 142)

[0745] NaH (60% oil solution) (121 mg, 2.77 mmol) was added to a stirred solution of 1-methylpiperidin-4-ol (218 mg, 1.890 mmol) in tetrahydrofuran (10 mL). The resulting mixture was stirred at 60 °C for 1 hour. Then, 2-allyl-6-((4-chlorophenyl)amino)-1-(6-fluoropyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (6,500 mg, 1.260 mmol) was added and the mixture was stirred at 60 °C for 16 hours. The reaction progress was monitored by TLC. After the reaction was complete, the reaction mixture was diluted with ice water (150 mL) and extracted with 10% methanol-DCM (2 x 100 mL). The combined organic layers were dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The crude material was purified by rapid chromatography (SiO2 / 230-400 mesh, 20-25% methanol-DCM) to obtain 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)oxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (175 mg, 0.352 mmol, 27.9% yield), which was a grayish-white solid.

[0746] Example 120. 2-Allyl-6-(4-fluoro-3-toluidine)-1-[6-(1-methyl-4-piperidinylamino)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 140)

[0747] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one. Yield: 129 mg, 75%.

[0748] Example 121. 2-Allyl-1-[6-(1-methyl-4-piperidinylamino)-2-pyridyl]-6-[p-(2,2,2-trifluoroethoxy)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 139)

[0749] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one. Yield: 80 mg, 66.6%.

[0750] Example 122. 2-Allyl-6-(p-chlorophenylamino)-1-[6-(1-methyl-4-piperidinylamino)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 138)

[0751] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one. Yield: 7.4 mg, 63%.

[0752] Example 123. 2-Allyl-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridinyl]-6-(1-methyl-4-pyrazolylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 137)

[0753] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 20 mg, 17%.

[0754] Example 124. 2-Allyl-6-(4-chloro-3-toluidine)-1-[6-(4-piperidinylamino)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 133)

[0755] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using 4-chloro-3-methylaniline and 4-({6-[6-(methylthioalkyl)-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]pyridin-2-yl}amino)piperidin-1-carboxylic acid tert-butyl ester. Yield: 24 mg, 38%.

[0756] Example 125. 2-Ethyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-6-(1-methyl-4-pyrazolylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 132)

[0757] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using 1-methyl-1H-pyrazol-4-amine and 2-ethyl-1-{6-[(1-methylpiperidin-4-yl)oxy]pyridin-2-yl}-6-(methylthioalkyl)-1H,2H,3H-pyrazol[3,4-d]pyrimidin-3-one. Yield: 35 mg, 56%.

[0758] Example 126. 2-Allyl-1-[6-(4-piperidinylamino)-2-pyridyl]-6-[p-(trifluoromethyl)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 131)

[0759] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one. Yield: 18 mg, 29%.

[0760] Example 127. 2-Allyl-1-[6-(4-piperidinylamino)-2-pyridyl]-6-[p-(2,2,2-trifluoroethoxy)phenylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 130)

[0761] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one. Yield: 6.9 mg.

[0762] Example 128. 2-Allyl-6-(4-fluoro-3-toluidine)-1-[6-(4-piperidinylamino)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 128)

[0763] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using 4-fluoro-3-methylaniline and 4-({6-[6-(methylthioalkyl)-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]pyridin-2-yl}amino)piperidin-1-carboxylic acid tert-butyl ester. Yield: 153 mg, 61%.

[0764] Example 129. 1-{6-[N-methyl(1-methyl-4-piperidinyl)amino]-2-pyridinyl}-2-ethyl-6-(1-methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 287)

[0765] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one. Yield: 35 mg, 60%.

[0766] Example 130. 2-Allyl-6-(4-methoxy-3-toluidine)-1-[6-(4-piperidinylamino)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 127)

[0767] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using 4-methoxy-3-methylaniline and 4-({6-[6-(methylthioalkyl)-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]pyridin-2-yl}amino)piperidin-1-carboxylic acid tert-butyl ester. Yield: 43 mg, 71%.

[0768] Example 131. 2-Allyl-1-[6-(4-piperidinylamino)-2-pyridyl]-6-m-toluidine-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 126)

[0769] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using 3-methylaniline and 4-({6-[6-(methylthioalkyl)-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]pyridin-2-yl}amino)piperidin-1-carboxylic acid tert-butyl ester. Yield: 39 mg, 67%.

[0770] Example 132. 2-Allyl-6-(p-chlorophenylamino)-1-[6-(4-piperidinylamino)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 124)

[0771] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using tert-butyl 4-({6-[6-(methylthioalkyl)-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]pyridin-2-yl}amino)piperidin-1-carboxylic acid (50 mg) and 4-chloroaniline (1 equivalent). Yield: 29 mg TFA salt (49%), as a yellow powder.

[0772] Example 133. 1-[6-(N-methyl-N-4-piperidinylamino)-2-pyridyl]-2-ethyl-6-(1-methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 286)

[0773] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using 1-methyl-1H-indazole-5-amine and 4-({6-[2-ethyl-6-(methylthioalkyl)-3-oxo-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]pyridin-2-yl}(methyl)amino)piperidin-1-carboxylic acid tert-butyl ester. Yield: 70 mg, 54%.

[0774] Example 134. 2-Methyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 121)

[0775] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using 2-methyl-1-{6-[(1-methylpiperidin-4-yl)oxy]pyridin-2-yl}-6-(methylthioalkyl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (69 mg) and 1-methyl-1H-indazole-5-amine (1 equivalent). Yield: 49 mg TFA salt (46%), as a yellow powder.

[0776] Example 135. 1-{6-[N-methyl(1-methyl-4-piperidinyl)amino]-2-pyridinyl}-2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 118)

[0777] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one. Yield: 9.8 mg, 68%.

[0778] Example 136. 6-Aniline-2-ethyl-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 117)

[0779] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using 2-ethyl-1-{6-[(1-methylpiperidin-4-yl)oxy]pyridin-2-yl}-6-(methylthioalkyl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (102 mg) and aniline (1 equivalent). Yield: 69 mg TFA salt (48%), as a yellow powder.

[0780] Example 137. 1-[6-(N-methyl-N-4-piperidinylamino)-2-pyridyl]-2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 116)

[0781] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using 1-methyl-1H-indazole-5-amine and 4-[methyl({6-[6-(methylthioalkyl)-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]pyridin-2-yl})amino]piperidin-1-carboxylic acid tert-butyl ester. Yield: 22 mg, 40%.

[0782] Example 138. 2-Ethyl-6-(1-Methyl-1H-indazol-5-ylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 114)

[0783] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using 2-ethyl-1-{6-[(1-methylpiperidin-4-yl)oxy]pyridin-2-yl}-6-(methylthioalkyl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (103 mg) and 1-methyl-1H-indazole-5-amine (2.5 equivalents). Yield: 64 mg TFA salt (41%), as a yellow powder.

[0784] Example 139. 2-Allyl-6-(2-methyl-4-pyridinylamino)-1-[6-(4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 113)

[0785] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridinylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one. Yield: 2 mg.

[0786] Example 140. 1-{6-[(S)-1-methyl-3-pyrrolylamino]-2-pyridyl}-2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (Compound 108)

[0787] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one. Yield: 6.5 mg TFA salt (24%), as a yellow powder.

[0788] Example 141. Synthesis of 1-{6-[(S)-3-pyrrolidinylamino]-2-pyridyl}-2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 107)

[0789] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one. Yield: 73 mg TFA salt (66%), as a yellow powder.

[0790] Example 142. Synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[6-(4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 106)

[0791] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, using tert-butyl piperidinium-1-carboxylate (184 mg) and 1-methyl-1H-indazole-5-amine (1 equivalent). Yield: 140 mg TFA salt (62%), as a yellow powder.

[0792] Example 143. Synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 109)

[0793] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, using 6-[(1-methyl-1H-indazol-5-yl)amino]-1-[6-(piperidin-4-yloxy)pyridin-2-yl]-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetate (15 mg). Yield: 9 mg TFA salt (60%), as a pale yellow powder.

[0794] Example 144. Synthesis of 1-{6-[(S)-1-methyl-3-pyrrolidinoxy]-2-pyridyl}-2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 105)

[0795] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, with the aid of (3S)-3-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester. Yield: 10.4 mg, 62%.

[0796] Example 145. Synthesis of 1-{6-[(S)-3-piperidinyloxy]-2-pyridyl}-2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 104)

[0797] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, using 1-methyl-1H-indazole-5-amine (1 equivalent) and (3S)-3-({6-[6-(methylthioalkyl)-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyridin-1-yl]oxy)piperidin-1-carboxylic acid tert-butyl ester (135 mg) prepared from (3S)-3-hydroxypiperidin-1-carboxylic acid tert-butyl ester. Yield: 92 mg TFA salt (56%), as a yellow powder.

[0798] Example 146. Synthesis of 1-{6-[(R)-3-pyrrolidinoxy]-2-pyridyl}-2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 101)

[0799] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, using 1-methyl-1H-indazole-5-amine and (3R)-3-({6-[6-(methylthioalkyl)-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]oxy)pyrrolidine-1-carboxylic acid tert-butyl ester prepared from (3R)-3-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester. Yield: 8.1 mg, 21%.

[0800] Example 147. Synthesis of 1-{6-[(S)-3-pyrrolidinoxy]-2-pyridyl}-2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 100)

[0801] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, using 1-methyl-1H-indazole-5-amine and (3S)-3-({6-[6-(methylthioalkyl)-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]oxy)pyrrolidine-1-carboxylic acid tert-butyl ester prepared from (3S)-3-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester. Yield: 55.1 mg, 60%.

[0802] Example 148. Synthesis of 2-allyl-6-(1-methyl-1H-1,3-benzimidazol-5-ylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 211)

[0803] Under nitrogen atmosphere, at room temperature, 41.5 mg (assumed 0.241 mmol) of mCPBA (<77% pure) in DCM (0.5 mL) was added to a stirred solution of 4-{6-[2-allyl-6-(methylthio)-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazaindene-1-yl]-2-pyridyloxy}-1-piperidinecarboxylic acid tert-butyl ester (100 mg, 0.201 mmol) in DCM (2.5 mL). The reaction was controlled by LC-MS. After 15 minutes, DCM was removed under vacuum, and the crude product was dissolved in MeCN (3 mL). Then, 1-methyl-1H-1,3-benzimidazol-5-ylamine (29.5 mg, 0.201 mmol) and methanesulfonic acid (26 µL, 0.401 mmol) were added. The reaction mixture was stirred in a sealed vial at 60 °C. After 18 hours, the reaction mixture was cooled to room temperature, and mCPBA quenched with 1M NaOH (5 mL) was added dropwise. The aqueous phase was extracted with EtOAc (3 x 20 mL) and then with brine (20 mL). The combined organic layers were dried (MgSO4) and concentrated under reduced pressure to give the product as a yellow powder. The crude material was dissolved in DCM (5 mL), loaded onto a 10 g silica gel column, and purified by rapid chromatography (0-100%, EtOAc: PE) to give the Boc-protected compound [85 mg, 94%] as a pale yellow solid. The DCM (5 mL) solution of this material (85 mg, 0.131 mmol) was treated with TFA (2.5 mL) for 2 hours. The solvent was removed under vacuum, and the crude product was dissolved in EtOAc, washed with saturated aqueous NaHCO3 solution and brine, dried (MgSO4), and concentrated. The obtained material was purified by reversed-phase chromatography (Gemini NX-C18, 21*150 mm, water (0.1% TFA) / acetonitrile, gradient over 12 minutes, 25 ml / min). The purified fractions were collected and concentrated to give 2-allyl-6-(1-methyl-1H-1,3-benzimidazol-5-ylamino)-1-[6-(4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, as trifluoroacetate, as a white solid [85 mg].

[0804] Example 149. Synthesis of 2-allyl-6-((1-methyl-1H-indol-5-yl)amino)-1-(6-((1-methylpiperidin-4-yl)oxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 186)

[0805] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, using 1-{6-[(1-methylpiperidin-4-yl)oxy]pyridin-2-yl}-6-(methylthioalkyl)-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (100 mg) and 1-methyl-1H-indole-5-amine (1.5 equivalents). Yield: 77 mg TFA salt (51%), as a powder.

[0806] Example 150. Synthesis of 1-{6-[(R)-3-pyrrolidinylamino]-2-pyridyl}-2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 102)

[0807] (3R)-3-[(6-bromopyridin-2-yl)amino]pyrrolidine-1-carboxylic acid tert-butyl ester

[0808] 2-Bromo-6-fluoropyridine (0.37 g, 2.1 mmol), DIPEA (6.3 mmol, 1.1 ml), and (3R)-3-aminopyrrolidine-1-carboxylic acid tert-butyl hydrochloride (0.52 g, 2.1 mmol) were mixed in 10 ml of DMSO. The reaction mixture was stirred at 100 °C for two days and partitioned between ethyl acetate and water. The organic phase was washed with water, saturated NaHCO3, and brine, dried over MgSO4, filtered, and concentrated. The residue was slurried with a mixture of heptane and ethyl acetate and collected by filtration to give 0.42 g (58%) (3R)-3-[(6-bromopyridin-2-yl)amino]pyrrolidine-1-carboxylic acid tert-butyl ester.

[0809] Step 2

[0810] Under nitrogen atmosphere, 6-(methylthioalkyl)-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (56 mg, 0.25 mmol), copper iodide (I) (52 mg, 0.27 mmol), potassium carbonate (104 mg, 0.75 mmol), 1,2-dimethylethylenediamine (44 mg, 0.50 mmol), and (3R)-3-[(6-bromopyridin-2-yl)amino]pyrrolidine-1-carboxylic acid tert-butyl ester (86 mg, 0.25 mmol) were mixed in 20 mL of dioxane. The reaction mixture was stirred overnight at 90 °C, diluted with ethyl acetate, filtered through diatomaceous earth, and concentrated. The residue was purified by rapid chromatography (silica, petroleum ether solution of 10-40% ethyl acetate).

[0811] 1-{6-[(R)-3-pyrrolidinylamino]-2-pyridyl}-2-allyl-6-(1-methyl-1H-indazole-5-ylamino) 1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one

[0812] The compound from step 2 was mixed with MCPBA (77%, 31 mg, 0.14 mmol) in 5 mL of DCM. After two hours, 1-methyl-1H-indazole-5-amine (37 mg, 0.25 mmol) was added. The reaction mixture was stirred overnight at room temperature. TFA (2 mL) was added, and the solvent was removed under reduced pressure after 3 hours. The residue was dissolved in methanol / water and purified by reversed-phase chromatography (Gemini NX-C18, 21*150 mm, water (0.1% TFA) / acetonitrile, gradient over 12 min, 25 mL / min). The purified fractions were collected and concentrated to give 13 mg (7% from two steps) of the title compound.

[0813] Example 151. Synthesis of 6-[(1-methyl-1H-indazol-5-yl)amino]-1-{6-[(3R)-piperidin-3-yloxy]pyridin-2-yl}-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one; bis(trifluoroacetic acid) (compound 103)

[0814] This compound was prepared using a method similar to that described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, using 1-methyl-1H-indazole-5-amine and (3R)-3-({6-[6-(methylthioalkyl)-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]pyridin-2-yl}oxy)piperidin-1-carboxylic acid tert-butyl ester. Yield: 130 mg, 56%.

[0815] Example 152. Synthesis of 6-[(1-methyl-1H-indazol-5-yl)amino]-1-[6-(piperidin-4-yloxy)pyridin-2-yl]-2-propyl-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 110)

[0816] To a solution of 6-[(1-methyl-1H-indazol-5-yl)amino]-1-[6-(piperidin-4-yloxy)pyridin-2-yl]-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid (60 mg, 0.098 mmol) in ethanol (5 mL), 10% palladium / carbon (4.4 mg) was added, and the mixture was stirred overnight at room temperature under a hydrogen atmosphere (1 atm). The solution was filtered through diatomaceous earth to remove the catalyst, and the diatomaceous earth pad was washed with methanol. The filtrate was concentrated under reduced pressure to give a crude product as a colorless oil. The crude product was purified by reversed-phase chromatography, and the purified fractions were collected and lyophilized to give the title compound. Yield: 20 mg TFA salt (33%).

[0817] Example 153. Synthesis of 6-[(1-methyl-1H-indazol-5-yl)amino]-1-{6-[(1-methylpiperidin-4-yl)oxy]pyridin-2-yl}-2-propyl-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 111)

[0818] To a solution of 6-[(1-methyl-1H-indazol-5-yl)amino]-1-{6-[(1-methylpiperidin-4-yl)oxy]pyridin-2-yl}-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid (41 mg, 0.066 mmol) in ethanol (4 mL), 10% palladium / carbon (2.95 mg) was added, and the mixture was stirred overnight at room temperature under a hydrogen atmosphere (1 atm). The solution was filtered through diatomaceous earth to remove the catalyst, and the diatomaceous earth pad was washed with methanol. The filtrate was concentrated under reduced pressure to give a crude product, a colorless oil, which was dissolved in acetonitrile and water and lyophilized to give the title compound. Yield: 33 mg TFA salt (80%).

[0819] Example 154. Synthesis of 1-{6-[(1-methylpiperidin-4-yl)oxy]pyridin-2-yl}-6-[(2-methylpyridin-4-yl)amino]-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 112).

[0820] This compound was prepared using a method similar to that described in the synthesis of 6-[(1-methyl-1H-indazol-5-yl)amino]-1-{6-[(1-methylpiperidin-4-yl)oxy]pyridin-2-yl}-2-propyl-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one, employing 6-[(2-methylpyridin-4-yl)amino]-1-[6-(piperidin-4-yloxy)pyridin-2-yl]-2-(propyl-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one. Yield: 31 mg TFA salt (28%), as a powder.

[0821] Example 155. Synthesis of 1-{6-[(R)-1-methyl-3-pyrrolidinoxy]-2-pyridyl}-2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 115)

[0822] 2-Allyl-1-(6-fluoropyridin-2-yl)-6-(methylsulfinyl)-1,2-dihydro-3H-pyrazolo[3,4-d] Pyrimidin-3-one

[0823] To a stirred solution of 2-allyl-1-(6-fluoropyridin-2-yl)-6-(methylthio)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (1 g, 3.15 mmol) in DCM (10 ml), m-CPBA (1.088 g, 6.30 mmol) was added, and the resulting reaction mixture was stirred at room temperature for 3 hours. The reaction progress was monitored by TLC. After the reaction was complete, the reaction mixture was quenched with sodium bicarbonate and extracted with DCM to give a crude product (1 g, 1.920 mmol, 60.9% yield), which was used for the next step.

[0824] 2-Allyl-1-(6-Fluoropyridin-2-yl)-6-((1-Methyl-1H-indazol-5-yl)amino)-1,2-dihydro-3H- Pyrazolo[3,4-d]pyrimidin-3-one

[0825] To a stirred solution of 2-allyl-1-(6-fluoropyridin-2-yl)-6-(methanesulfonyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (1 g, 2.86 mmol) in acetic acid (10 mL), 1-methyl-1H-indazole-5-amine (3,0.421 g, 2.86 mmol) was added. The resulting reaction mixture was stirred at room temperature for 16 hours. The reaction progress was monitored by TLC. After the reaction was complete, the reaction mixture was concentrated and quenched with ice-cold water. The resulting solid was filtered and washed with MTBE (20 mL) to give the pure compound 2-allyl-1-(6-fluoropyridin-2-yl)-6-((1-methyl-1H-indazol-5-yl)amino)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (800 mg, 1.767 mmol, 61.7% yield, as a brown solid).

[0826] (R)-2-Allyl-6-((1-Methyl-1H-indazol-5-yl)amino)-1-(6-((1-Methylpyrrolidone-3-yl)) (Oxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one

[0827] At 60 °C, NaH (60% oil solution) (214 mg, 8.90 mmol) was added to a stirred solution of (R)-1-methylpyrrolidine-3-ol (5,300 mg, 2.97 mmol) in THF (5 ml) and stirred for 1 hour. Then, 2-allyl-1-(6-fluoropyridin-2-yl)-6-((1-methyl-1H-indazol-5-yl)amino)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (4,300 mg, 0.720 mmol) was added to the above reaction mixture and stirred at room temperature for 16 hours. The reaction progress was monitored by TLC after completion. The reaction was quenched with ice-cold water and extracted with DCM to obtain a crude product, which was concentrated under reduced pressure and then subjected to preparative HPLC (X-SELECT C18 150M, 0.1% FA IN H2O). After lyophilization, the fractions were collected to give pure (R)-2-allyl-6-((1-methyl-1H-indazol-5-yl)amino)-1-(6-((1-methylpyrrolidone-3-yl)oxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (90 mg, 0.179 mmol, 6.04% yield).

[0828] Example 156. Synthesis of 1-(6-{[(3R)-1-ethylpyrrolidine-3-yl]amino}pyridin-2-yl)-6-[(1-methyl-1H-indazol-5-yl)amino]-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 119)

[0829] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, using 6-[(1-methyl-1H-indazol-5-yl)amino]-2-(prop-2-en-1-yl)-1-(6-{[(3R)-pyrrolidine-3-yl]amino}pyridin-2-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid (45 mg) and acetaldehyde (excess). Yield: 15 mg TFA salt (32%), as a powder.

[0830] Example 157. Synthesis of 6-[(1-methyl-1H-indazol-5-yl)amino]-2-(prop-2-en-1-yl)-1-(6-{[(3R)-1-(propane-2-yl)pyrrolidine-3-yl]amino}pyridin-2-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 120)

[0831] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidinoxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, using 6-[(1-methyl-1H-indazol-5-yl)amino]-2-(prop-2-en-1-yl)-1-(6-{[(3R)-pyrrolidine-3-yl]amino}pyridin-2-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid (45 mg) and acetone (excess). Yield: 30 mg TFA salt (63%), as a powder.

[0832] Example 158. Synthesis of 6-[(1-methyl-1H-indazol-5-yl)amino]-1-{6-[(piperidin-4-yl)amino]pyridin-2-yl}-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 122)

[0833] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(piperidin-4-ylamino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using tert-butyl piperidin-1-carboxylate (182 mg) and 1-methyl-1H-indazole-5-amine (1 equivalent). Yield: 107 mg TFA salt (48%), as a yellow powder.

[0834] Example 159. Synthesis of 6-(phenylamino)-1-{6-[(piperidin-4-yl)amino]pyridin-2-yl}-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 123)

[0835] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(piperidin-4-ylamino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using tert-butyl 4-({6-[6-(methylthioalkyl)-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]pyridin-2-yl}amino)piperidin-1-carboxylic acid (50 mg) and aniline (3 equivalents). Yield: 31 mg TFA salt (55%), as a yellow powder.

[0836] Example 160. Synthesis of 6-[(4-fluorophenyl)amino]-1-{6-[(piperidin-4-yl)amino]pyridin-2-yl}-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 125)

[0837] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(piperidin-4-ylamino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using tert-butyl 4-({6-[6-(methylthioalkyl)-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]pyridin-2-yl}amino)piperidin-1-carboxylic acid (50 mg) and 4-fluoroaniline (2 equivalents). Yield: 37 mg TFA salt (64%), as a powder.

[0838] Example 161. Synthesis of 6-[(1-methyl-1H-indazol-5-yl)amino]-1-{6-[(1-methylpiperidin-4-yl)amino]pyridin-2-yl}-2-(propen-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 129)

[0839] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using 6-[(1-methyl-1H-indazol-5-yl)amino]-1-{6-[(piperidin-4-yl)amino]pyridin-2-yl}-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid (38 mg). Yield: 20 mg (52%), as a powder.

[0840] Example 162. Synthesis of 2-methyl-4-[(3-oxo-1-{6-[(piperidin-4-yl)amino]pyridin-2-yl}-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-6-yl)amino]benzyl nitrile (compound 134)

[0841] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridinylamino)-1-[6-(1-methyl-4-piperidinyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one, using tert-butyl 4-[(6-{6-[(4-cyano-3-methylphenyl)amino]-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl}pyridin-2-yl)oxy]piperidin-1-carboxylic acid, followed by deprotection of the boc group. Yield: 15.3 mg TFA salt (39%), as a white powder.

[0842] Example 163. Synthesis of 1-{6-[(piperidin-4-yl)amino]pyridin-2-yl}-2-(prop-2-en-1-yl)-6-{[4-(trifluoromethoxy)phenyl]amino}-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 135)

[0843] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using tert-butyl 4-({6-[6-(methylthioalkyl)-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]pyridin-2-yl}amino)piperidin-1-carboxylic acid (50 mg) and 4-(trifluoromethoxy)aniline (2 equivalents). Yield: 27 mg (51%), as a powder.

[0844] Example 164. Synthesis of 6-{[3-methyl-4-(trifluoromethoxy)phenyl]amino}-1-{6-[(piperidin-4-yl)amino]pyridin-2-yl}-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 136)

[0845] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using tert-butyl 4-({6-[6-(methylthioalkyl)-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]pyridin-2-yl}amino)piperidin-1-carboxylic acid (50 mg) and 3-methyl-4-(trifluoromethoxy)aniline (2 equivalents). Yield: 29 mg TFA salt (44%), as a powder.

[0846] Example 165. Synthesis of 6-[(4-bromophenyl)amino]-1-{6-[(piperidin-4-yl)amino]pyridin-2-yl}-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 143)

[0847] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using tert-butyl 4-({6-[6-(methylthioalkyl)-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]pyridin-2-yl}amino)piperidin-1-carboxylic acid (60 mg) and 4-bromoaniline (2 equivalents). Yield: 27.5 mg (44%), as a white powder.

[0848] Example 166. Synthesis of 6-[(3-bromo-5-chlorophenyl)amino]-1-{6-[(piperidin-4-yl)amino]pyridin-2-yl}-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 144)

[0849] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using tert-butyl 4-({6-[6-(methylthioalkyl)-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]pyridin-2-yl}amino)piperidin-1-carboxylic acid (50 mg), 3-bromo-5-chloroaniline (2 equivalents), and methanesulfonic acid (1 equivalent). Yield: 25 mg TFA salt (37%), as a powder.

[0850] Example 167. Synthesis of 6-[(4-chlorophenyl)amino]-1-{6-[(1-methylpiperidin-4-yl)(propyl)amino]pyridin-2-yl}-2-(propyl-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 149)

[0851] 4-[(6-bromopyridin-2-yl)(propyl)amino]piperidine-1-carboxylic acid tert-butyl ester

[0852] At 0 °C, a solution of 60% sodium hydride in mineral oil (113 mg, 2.82 mmol, 1.3 equivalents) was added to a stirred solution of 4-[(6-bromopyridin-2-yl)amino]piperidine-1-carboxylic acid tert-butyl ester (773 mg, 2.17 mmol, 1 equivalent) and propyl methanesulfonate (360 mg, 2.6 mmol, 1.2 equivalents) in DMF (8 mL). After complete addition, the mixture was brought to room temperature and stirred for 40 min. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (3 × 120 mL). The organic layer was washed with water (4 × 100 mL) and brine (100 mL), and then dried over magnesium sulfate. The organic layer was concentrated to the residue under reduced pressure. The residue was purified by rapid silica chromatography, eluting with a gradient of 5-70% ethyl acetate in petroleum ether, to give the title compound (470 mg, 54%) as a colorless oil.

[0853] 4‐({6‐[6‐(methylthioalkyl)‐3‐oxo‐2‐(propyl‐2‐en‐1‐yl)‐1H,2H,3H‐pyrazolo[3,4‐d] ] ] [Pyrimidin-1-yl]pyridin-2-yl}(propyl)amino)piperidin-1-carboxylic acid tert-butyl ester

[0854] This intermediate was prepared using a method similar to that described in the synthesis of tert-butyl piperidine-1-carboxylate, with 4-[(6-bromopyridin-2-yl)(propyl)amino]piperidine-1-carboxylate (475 mg). Yield: 519 mg (81%).

[0855] 6-[(4-chlorophenyl)amino]-1-{6-[(1-methylpiperidin-4-yl)(propyl)amino]pyridin-2-yl}-2- (propyl-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one

[0856] The title compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using 6-[(4-chlorophenyl)amino]-1-{6-[(piperidin-4-yl)(propyl)amino]pyridin-2-yl}-2-(propyl-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid (85 mg). Yield: 65 mg TFA salt (75%), as a powder.

[0857] Example 168. Synthesis of 6-{[3-methyl-5-(trifluoromethoxy)phenyl]amino}-1-{6-[(piperidin-4-yl)amino]pyridin-2-yl}-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 155)

[0858] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using tert-butyl 4-({6-[6-(methylthioalkyl)-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]pyridin-2-yl}amino)piperidin-1-carboxylic acid (100 mg) and 3-methyl-5-(trifluoromethoxy)aniline (1.5 equivalents). Yield: 60 mg TFA salt (46%), as a powder.

[0859] Example 169. Synthesis of 6-[(4-chlorophenyl)amino]-1-{6-[methyl(piperidin-4-yl)amino]pyridin-2-yl}-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 156)

[0860] This compound was prepared using a method similar to that described in the synthesis of 6-[(4-chlorophenyl)amino]-1-{6-[(1-methylpiperidin-4-yl)(propyl)amino]pyridin-2-yl}-2-(propen-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one, using tert-butyl 4-[methyl({6-[6-(methylthioalkyl)-3-oxo-2-(propen-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl})amino]piperidin-1-carboxylic acid (200 mg) and 4-chloroaniline (1 equivalent). Yield: 112 mg (58%), as a white powder.

[0861] Example 170. Synthesis of 6-[(4-chlorophenyl)amino]-1-{6-[methyl(1-methylpiperidin-4-yl)amino]pyridin-2-yl}-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 157)

[0862] This compound was prepared using a method similar to that described in the synthesis of 6-[(4-chlorophenyl)amino]-1-{6-[(1-methylpiperidin-4-yl)(propyl)amino]pyridin-2-yl}-2-(propyl-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one, using 6-[(4-chlorophenyl)amino]-1-{6-[methyl(piperidin-4-yl)amino]pyridin-2-yl}-2-(propyl-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (65 mg). Yield: 68 mg TFA salt (83%), as a pale yellow powder.

[0863] Example 171. Synthesis of 6-[(4-chlorophenyl)amino]-1-{6-[(3R)-piperidin-3-yloxy]pyridin-2-yl}-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 159)

[0864] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using tert-butyl 4-({6-[6-(methylthioalkyl)-3-oxo-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl]pyridin-2-yl}amino)piperidin-1-carboxylic acid (166 mg) and 4-chloroaniline (1 equivalent). Yield: 150 mg TFA salt (76%), as a yellow powder.

[0865] Example 172. Synthesis of 6-[(4-chlorophenyl)amino]-1-(6-{[(3S)-1-methylpiperidin-3-yl]oxy}pyridin-2-yl)-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 162)

[0866] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using 6-[(4-chlorophenyl)amino]-1-{6-[(3R)-piperidin-3-yloxy]pyridin-2-yl}-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid (kdh-0088) (150 mg). Yield: 82 mg free base (53%), in powder form.

[0867] Example 173. Synthesis of 1-{6-[methyl(piperidin-4-yl)amino]pyridin-2-yl}-2-(propen-2-yl)-6-{[1-(propane-2-yl)-1H-pyrazol-4-yl]amino}-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 169)

[0868] This compound was prepared using a method similar to that described in the synthesis of 6-[(4-chlorophenyl)amino]-1-{6-[(1-methylpiperidin-4-yl)(propyl)amino]pyridin-2-yl}-2-(propen-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one, using 4-[methyl({6-[6-(methylthioalkyl)-3-oxo-2-(propen-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl})amino]piperidin-1-carboxylic acid tert-butyl ester (200 mg) and 1-isopropyl-4-pyrazolamine (1.5 equivalents). Yield: 139 mg TFA salt (59%), as a powder.

[0869] Example 174. Synthesis of 1-{6-[methyl(piperidin-4-yl)amino]pyridin-2-yl}-6-{[1-(2-methylpropyl)-1H-pyrazol-4-yl]amino}-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 170)

[0870] This compound was prepared using a method similar to that described in the synthesis of 6-[(4-chlorophenyl)amino]-1-{6-[(1-methylpiperidin-4-yl)(propyl)amino]pyridin-2-yl}-2-(propen-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one, using 4-[methyl({6-[6-(methylthioalkyl)-3-oxo-2-(propen-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl})amino]piperidin-1-carboxylic acid tert-butyl ester (200 mg) and 1-isobutyl-4-pyrazolamine (1.5 equivalents). Yield: 153 mg TFA salt (63%), as a powder.

[0871] Example 175. Synthesis of 1-{6-[methyl(1-methylpiperidin-4-yl)amino]pyridin-2-yl}-2-(propen-2-yl)-6-{[1-(propane-2-yl)-1H-pyrazol-4-yl]amino}-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 174)

[0872] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using 1-{6-[methyl(piperidin-4-yl)amino]pyridin-2-yl}-2-(prop-2-en-1-yl)-6-{[1-(propane-2-yl)-1H-pyrazol-4-yl]amino}-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid (110 mg). Yield: 37 mg TFA salt (33%), as a powder.

[0873] Example 176. Synthesis of 1-{6-[methyl(1-methylpiperidin-4-yl)amino]pyridin-2-yl}-6-{[1-(2-methylpropyl)-1H-pyrazol-4-yl]amino}-2-(propyl-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 175)

[0874] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using 1-{6-[methyl(piperidin-4-yl)amino]pyridin-2-yl}-6-{[1-(2-methylpropyl)-1H-pyrazol-4-yl]amino}-2-(propyl-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one; trifluoroacetic acid (126 mg). Yield: 67 mg TFA salt (52%), as a powder.

[0875] Example 177. Synthesis of 1-{6-[(3R)-1-azabicyclo[2.2.2]octyl-3-yloxy]pyridin-2-yl}-6-[(4-chlorophenyl)amino]-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 176)

[0876] This compound was prepared using a method similar to that described in the synthesis of (R)-2-allyl-6-((1-methyl-1H-pyrazol-3-yl)amino)-1-(6-(quininecyclo-3-yloxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using 1-{6-[(3R)-1-azabicyclo[2.2.2]octyl-3-yloxy]pyridin-2-yl}-6-(methylthioalkyl)-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazololo[3,4-d]pyrimidin-3-one (100 mg) and 4-chloroaniline (2 equivalents). Yield: 67 mg TFA salt (46%), as a yellow powder.

[0877] Example 178. Synthesis of 2-allyl-1-[6-(4-piperidinyloxy)-2-pyridinyl]-6-(1-propyl-4-pyrazolylamino)-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 179)

[0878] This compound was prepared using a method similar to that described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-(((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, using 4-{6-[2-allyl-6-(methylthio)-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazaindene-1-yl]-2-pyridyloxy}-1-piperidinecarboxylic acid tert-butyl ester (123 mg) and 1-propyl-4-pyrazolylamine (48.9 mg). Yield: 12 mg TFA salt.

[0879] Example 179. Synthesis of 1-{6-[methyl(piperidin-4-yl)amino]pyridin-2-yl}-6-[(1-methyl-1H-pyrazol-4-yl)amino]-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (compound 182).

[0880] This compound was prepared using a method similar to that described in the synthesis of 6-[(4-chlorophenyl)amino]-1-{6-[(1-methylpiperidin-4-yl)(propyl)amino]pyridin-2-yl}-2-(propen-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one, using 4-[methyl({6-[6-(methylthioalkyl)-3-oxo-2-(propen-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-1-yl})amino]piperidin-1-carboxylic acid tert-butyl ester (200 mg) and 1-methyl-4-pyrazolamine (1 equivalent) to give a crude free base intermediate (227 mg) as a brown oil, of which 13% was purified by preparative HPLC. Yield: 13.4 mg TFA salt (45%), in powder form.

[0881] Example 180. Synthesis of 1-[6-(N-methyl-N-4-piperidinylamino)-2-pyridyl]-2-allyl-6-[1-(2-fluoro-2-methylpropyl)-4-pyrazolylamino]-1,2-dihydro-3H-1,2,5,7-tetraazaindene-3-one (compound 183)

[0882] This compound was prepared using a method si...

Claims

1. A compound having the structural formula AA: (AA), Or its solvates, enantiomers, tautomers or diastereomers, or pharmaceutically acceptable salts of any of the foregoing substances, wherein: X, Y, and Z are each independently CH or N; R 1 It is -O-, -NH-, or -N(C1-C3 alkyl)-; Each R 2 Independently, it is a fluorinated group, -CN, unsubstituted C1-C5 alkyl, fluorinated C1-C5 alkyl, or cyano-substituted C1-C5 alkyl, wherein the two R groups bonded to the same carbon atom are... 2 Optionally, they can be combined to form a spirocyclic fused C3-C7 cycloalkyl ring, or two R atoms bonded to different carbon atoms. 2 Optionally, they can be combined to form a bridged or fused C3-C5 cycloalkyl ring, or an R 2 and R 8 They can be optionally combined to form bridging or fused 3- to 5-membered rings; R 3 It is a -C1-C3 alkyl group or -CH2-CH=CH2; R 4 yes , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or ,in" " indicates R 4 Connection points with compounds; Each R 5 and each R 6 Independently, it is hydrogen, a halogroup, -CN, a -C1-C4 alkyl group optionally substituted with a halogroup, an -O-(C1-C4 alkyl group optionally substituted with a halogroup, an -O-(C1-C4 alkyl group substituted with a C3-C6 cycloalkyl group, an N-linked saturated 3-7 membered heterocyclic group, a 5-6 membered heteroaryl group, or a phenyl group, wherein there are no more than two R groups. 6 It is not hydrogen; the R on the adjacent ring atoms 5 and R 6 Optionally together form with R 4 Fused 4-7 membered saturated heterocycles or cycloalkyl rings; R 7 It is hydrogen, -C1-C4 alkyl, -C1-C4 alkylene, -O-C1-C4 alkyl, -C(O)-C1-C4 alkyl, or C3-C6 cycloalkyl, wherein R 7 Any C1-C4 alkyl or C1-C4 alkylene moiety may optionally be substituted by one or more substituents independently selected from halogen and -CN; R 8 It is hydrogen, -C1-C4 alkyl, or 4-6 membered saturated heterocycle; R 9 It is hydrogen, a halogroup, -CN, a -C1-C4 alkyl group optionally substituted with a halogroup, an -O-C1-C4 alkyl group optionally substituted with a halogroup, or a phenyl group optionally substituted; m is 0, 1, 2, 3 or 4; n is 0, 1, 2, or 3; and m+n is 1, 2, 3, or 4. Any hydrogen atom may be optionally replaced by deuterium.

2. The compound of claim 1, wherein: X, Y, and Z are each independently CH or N; R 1 It is -O-, -NH-, or -N(C1-C3 alkyl)-; Each R 2 Independently, it is a fluorinated group, -CN, unsubstituted C1-C5 alkyl, fluorinated C1-C5 alkyl, or cyano-substituted C1-C5 alkyl, wherein the two R groups bonded to the same carbon atom are... 2 Optionally, they can be combined to form a spirocyclic fused C3-C7 cycloalkyl ring, or two R atoms bonded to different carbon atoms. 2 Optionally, they can be combined to form a bridged C3-C5 cycloalkyl ring, or an R 2 and R 8 They can be optionally combined to form bridging or fused 3- to 5-membered rings; R 3 It is a -C1-C3 alkyl group or -CH2-CH=CH2; R 4 yes , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or ,in" " indicates R 4 Connection points with compounds; Each R 5 and each R 6 Independently, it is hydrogen, a halogroup, -CN, a -C1-C4 alkyl group optionally substituted with a halogroup, or an -O-(C1-C4 alkyl group optionally substituted with a halogroup), wherein there are no more than two R groups. 6 It is not hydrogen; the R on the adjacent ring atoms 5 and R 6 Together with R 4 Fused 4-7 membered saturated heterocycles or cycloalkyl rings; R 7 It is hydrogen, -C1-C4 alkyl or -C1-C4 alkylene-O-C1-C4 alkyl, wherein R 7 Any C1-C4 alkyl or C1-C4 alkylene moiety may optionally be substituted by one or more substituents independently selected from halogen and -CN; R 8 It is hydrogen or -C1-C4 alkyl; R 9 It is hydrogen, a halogroup, -CN, a -C1-C4 alkyl group optionally substituted with a halogroup, an -O-C1-C4 alkyl group optionally substituted with a halogroup, or a phenyl group optionally substituted; m is 0 or 1; n is 0, 1, 2, or 3; and m and n are not both 0.

3. The compound according to claim 1 or 2, wherein: X, Y, and Z are each CH.

4. The compound according to claim 1 or 2, wherein: X is N and Y and Z are each CH.

5. The compound of claim 2, wherein: X and Y are both N and Z is CH.

6. The compound according to any one of claims 1-5, wherein R 4 yes: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or .

7. The compound of claim 6, wherein: R 4 yes: , or ; R 5 It is hydrogen, a halogroup, -CN, a C1-C4 alkyl group optionally substituted with a halogroup, or a -O-(C1-C4 alkyl group optionally substituted with a halogroup; and Each R 6 Independently, it may be hydrogen, a halogroup, -CN, a -C1-C4 alkyl group optionally substituted with a halogroup, an -O-(C1-C4 alkyl group optionally substituted with a halogroup, an -O-(C1-C4 alkyl group substituted with a C3-C6 cycloalkyl group, an N-linked saturated 3-7 membered heterocyclic group, a 5-6 membered heteroaryl group, or a phenyl group; or R bonded to adjacent ring atoms 5 and R 6 Together they form methylenedioxy groups.

8. The compound according to any one of claims 1-7, wherein: Each R 6 Independently, it is a hydrogen, fluorinated, brominated, chlorolated, -CN, -CF3, -CH3, -OCH3, -OCF3, -OCH2CF3, -OCH2CH3, -OCH2CH2F, -OCH2CH(CH3)CH3, cyclopropylmethoxy, morpholin-4-yl, thiomorpholin-4-yl, 1-methyl-1H-pyrazolyl, or a phenyl group optionally substituted with a halogroup; and Each R 5 It can be hydrogen, fluorinated, chloro, -CN, -CF3, -CH3, or -OCH3.

9. The compound according to any one of claims 1-8, wherein no more than one R 6 It's not hydrogen.

10. The compound of claim 6, wherein: R 4 yes: , , , , , or ; R 6 It is a hydrogen or chloride group; and R 7 It is hydrogen, -CH3, -CH2CH3, -CH2C(CH3)2F, -CH2CH2CF3, -CH2CH2CH3, -CH(CH3)2, -CH2CH(CH3)2, -CH2CH(CH3)2, -C(CH3)3, -CH2CH2CH2F, -OCH2CF3 or cyclopropyl.

11. The compound of claim 6, wherein: R 4 yes: , , , , , or ;and Each R 6 It is hydrogen or -CH3; and R 9 It is an optional substituted phenyl group.

12. The compound of claim 6, wherein: R 4 yes: or , where each R 6 It is hydrogen.

13. The compound of claim 6, wherein R 4 yes , or , where each R 6 It is either hydrogen or methyl.

14. The compound of claim 6, wherein R 4 yes , or , where each R 6 It is hydrogen; and R 7 When present, it is isopropyl.

15. The compound of claim 6, wherein R 4 yes , where R 7 It is methyl or -C(O)CH3 and each R 6 It is hydrogen.

16. The compound of claim 6, wherein: R 4 yes: , , , or , where each R 6 It is hydrogen.

17. The compound of claim 6, wherein R 4 It is phenyl, 3-methylphenyl, 3-chloro-5-bromophenyl, 3,4-dichlorophenyl, 4-bromophenyl, 4-chlorophenyl, 4-fluorophenyl, 4-(1-methylpiperidin-4-yl)-5-methylphenyl, 4-trifluoromethoxyphenyl, 3-methyl-4-trifluoromethoxyphenyl, 4-(2,2,2-trifluoroeth-1-yl)oxyphenyl, 3-methyl-4-(2,2,2-trifluoroeth-1-yl)oxyphenyl, 4-trifluoromethylphenyl, 3-methyl-4-chlorophenyl, 3-methyl-4-fluorophenyl, 3-methyl-5-fluorophenyl, 3-methyl-4-cyanophenyl, 3-methyl-4-methoxyphenyl, 3-methyl-4-(2,2,2-trifluoroeth-1-yloxy)phenyl, 3,4-methylene 4-(cyclopropylmethyloxy)phenyl, 4-(morpholin-4-yl)phenyl, 4-(thiomorpholin-4-yl)phenyl, 4-(2-methylpropyl-1-yloxy)phenyl, 3-(1-methyl-1H-pyrazol-4-yl)phenyl, 4-(1-methyl-1H-pyrazol-4-yl)phenyl, 3-methyl-4-(morpholin-4-yl)phenyl, 4-(phenyl)phenyl, 3-bromophenyl, 3-cyanophenyl, 4-(2-fluoroethoxy-1-yl)phenyl, 4-cyanophenyl, 4-(1,1-dioxothiazin-4-yl)phenyl, 3-(pyridin-3-yl)phenyl, 3-(pyrimidin-5-yl)phenyl, 3-(1H-imidazol-1-yl)phenyl, 3-(1H-pyrazol-1-yl)phenyl 3-(d3-methyl)phenyl, 4-(3-fluoroprop-1-yloxy)phenyl, 4-ethoxyphenyl, 2-methylpyridin-4-yl, 6-methylpyridin-3-yl, 2,6-dimethylpyridin-4-yl, 2-trifluoromethyl-6-methylpyridin-4-yl, 2-trifluoromethylpyridin-4-yl, pyridin-3-yl, 2-methylpyridin-5-yl, 2-methoxypyridin-4-yl, 2-methoxypyridin-5-yl, 3-chloropyridin-5-yl, 3-fluoropyridin-5-yl, 3-cyanopyridin-5-yl, 3-methylpyridin-5-yl, 3-methoxypyridin-5-yl, 1-methyl-1H-pyrazol-4-yl, 1-propyl-1H-pyrazol-4-yl, 1-isopropyl-1H-pyrazol-4-yl 1-(2,2-dimethylethyl-1-yl)pyrazol-4-yl, 1-(2-fluoro-2,2-dimethylethyl-1-yl)-1H-pyrazol-4-yl, 1-(3,3,3-trifluoroprop-1-yl)-1H-pyrazol-4-yl, 1H-indazole-5-yl, 1-methyl-1H-indazole-5-yl, 1-ethyl-1H-indazole-5-yl, 1-propyl-1H-indazole-5-yl, 1-(3-fluoroprop-1-yl)-1H-indazole-5-yl, 1-isopropyl-1H-indazole-5-yl, 1-isobutyl-1H-indazole-5-yl, 1-cyclopropyl-1H-indazole-5-yl, 3-chloro-1H-indazole-5-yl, 3-methyl-1H-indazole-5-yl, 1,3-Dimethyl-1H-indazole-5-yl, 1H-indazole-4-yl, 2-methyl-2H-indazole-5-yl, 2-ethyl-2H-indazole-5-yl, 3-phenylisothiazol-5-yl, benzofuran-6-yl, benzofuran-5-yl, 1-methyl-1H-indole-5-yl, 1-acetyl-1H-indole-5-yl, 1,2,4-triazolo[1,5-a]pyridin-6-yl, quinoxaline-6-yl, quinoline-6-yl, quinoline-7-yl, isoquinoline-6-yl, isoquinoline-7-yl, 2-methylbenzo[d]oxazol-5-yl, 2-methylbenzo[d]oxazol-6-yl, benzo[d]isooxazol-6 -yl, benzo[d]thiazol-6-yl, 2-methylbenzo[d]thiazol-6-yl, 1-methyl-1H-benzo[d]imidazol-5-yl, benzo[d]thiazol-5-yl, 1-methyl-1H-indazole-6-yl, 2-methyl-2H-indazole-6-yl, imidazo[1,2-a]pyridin-6-yl, 2-(2-fluoroethoxy)pyridin-5-yl, 2-(tert-butyl)-2H-indazole-5-yl, 1-isopropyl-1H-benzo[d]imidazol-5-yl, 1-isopropyl-1H-pyrazolo[3,4-b]pyridin-5-yl or 1-isopropyl-1H-benzo[d][1,2,3]triazol-5-yl.

18. The compound according to any one of claims 1-17, wherein the structure is: The ring represented is selected from pyrrolidine-3-yl, 1-methylpyrrolidine-3-yl, 1-ethylpyrrolidine-3-yl, 1-isopropylpyrrolidine-3-yl, piperidin-4-yl, piperidin-3-yl, 1-methylpiperidin-4-yl, 1-methylpiperidin-3-yl, quinine-3-yl, 8-methyl-8-azabicyclo[3.2.1]octane-3-yl and 9-methyl-9-azabicyclo[3.3.1]nonane-3-yl.

19. The compound according to any one of claims 1-17, wherein the structure is: The ring represented is selected from pyrrolidine-3-yl, 1-methylpyrrolidine-3-yl, 1-ethylpyrrolidine-3-yl, 1-isopropylpyrrolidine-3-yl, piperidin-4-yl, piperidin-3-yl, 1-methylpiperidin-4-yl, 1-methylpiperidin-3-yl, 1-(methyl-d3)piperidin-4-yl, azircycloheptane-4-yl, quinine-3-yl, 8-methyl-8-azibicyclo[3.2.1]octane-3-yl and 9-methyl-9-azibicyclo[3.3.1]nonane-3-yl.

20. The compound according to any one of claims 1-19, wherein R 3 It is methyl, ethyl, n-propyl or -CH2-CH=CH2.

21. The compound of claim 20, wherein R 3 It is -CH2-CH=CH2.

22. The compound according to any one of claims 1-21, wherein R 1 It is -O-, -N(CH3)-, -N(CH2CH2CH3)- or -NH-.

23. The compound of claim 22, wherein R 1 Yes -O-.

24. The compound of claim 1, wherein the compound is any one of the following compounds: 。 25. The compound of claim 1, wherein the compound is any one of the following compounds: 。 26. A pharmaceutical composition comprising an effective amount of the compound according to any one of claims 1-25; and a pharmaceutically acceptable carrier.

27. A method for inhibiting Wee1A kinase activity in a subject, comprising administering to the subject an effective amount of a compound of any one of claims 1-25 or a composition of claim 26.

28. A method of treating a subject suffering from cancer or other cell growth disorders characterized by abnormal Wee1A kinase activity, comprising the step of administering to the subject an effective amount of a compound of any one of claims 1-25 or a composition of claim 26.

29. The method of claim 28, wherein the subject suffers from cancer associated with p53 inactivation.

30. A method for inhibiting both Wee1A kinase activity and Myt1 kinase activity in a subject, comprising administering to the subject an effective amount of a compound of any one of claims 1-25 as a dual inhibitor or a composition of claim 26 containing a dual inhibitor.

31. A method of treating a subject suffering from cancer or other cell growth disorders characterized by both abnormal Wee1A kinase activity and abnormal Myt1 kinase activity, comprising the step of administering to the subject an effective amount of a compound of any one of claims 1-25 or a composition of claim 26.

32. The method of any one of claims 28, 29, or 31, wherein the cancer is selected from brain cancer, cervical brain cancer, heart cancer, gastrointestinal cancer, esophageal cancer, thyroid cancer, small cell carcinoma, non-small cell carcinoma, breast cancer, lung cancer, gastric cancer, gallbladder / choleduct cancer, liver cancer, pancreatic cancer, colon cancer, rectal cancer, ovarian cancer, choriocarcinoma, endometrial cancer, cervical cancer, renal pelvis / ureter cancer, bladder cancer, prostate cancer, penile cancer, testicular cancer, fetal cancer, Wilms' disease, skin cancer, malignant melanoma, neuroblastoma, osteosarcoma, Ewing's tumor, soft tissue sarcoma, acute leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, polycythemia vera, malignant lymphoma, multiple myeloma, Hodgkin's lymphoma, and non-Hodgkin's lymphoma.

33. The method of claim 32, wherein the subject suffers from a cancer selected from serous uterine carcinoma or renal carcinoma.

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