Compounds, pharmaceutical compositions, methods for preparing compounds, and methods of using them

Abstract

This document discloses compounds that can be used to treat subjects in need, as well as pharmaceutically acceptable salts thereof. The compounds disclosed herein may be inhibitors of ataxia-telangiectasia and RAD-3-related protein kinase (ATR). Pharmaceutical compositions comprising said compounds or pharmaceutically acceptable salts thereof, and methods of their preparation and use, are also disclosed.

Description

Technical Field

[0001] This invention relates to compounds and pharmaceutical compositions, their preparation, and their use in the treatment of diseases or conditions (e.g., cancer) and, in particular, those diseases or conditions (e.g., cancer) that depend on the activity of ataxia-telangiectasia and RAD-3-associated protein (ATR) kinase. Background Technology

[0002] DNA damage persists in cells due to environmental damage, including ultraviolet radiation, X-rays, and endogenous stress factors such as reactive oxygen species and alkaline hydrolysis. Cancer cells experience a higher rate of DNA damage, which is essentially induced by the higher rate of DNA replication in these cells. Several DNA damage response (DDR) pathways evolve in a highly coordinated manner to help repair DNA damage and act as cellular checkpoints to stop DNA-damaged cell replication, allowing repair functions to occur before damaged DNA is passed on to daughter cells. Each identified DNA repair pathway can sense and repair different but overlapping types of DNA damage.

[0003] One major DDR protein acting as a key cell cycle checkpoint is the ataxia-telangiectasia mutant and rad3-associated (ATR) kinase, which is associated with the phosphatidylinositol 3-kinase-associated protein kinase (PIKK) family. ATR is activated by single-stranded (ss) DNA damage caused by stalled replication forks or during nucleotide excision repair, but also by double-strand breaks following DNA end excision during homologous recombination. ATR is recruited to DNA damage sites by binding to RPA proteins that coat ss DNA with a cofactor called an ATR interacting protein (ATRIP). The ATR / ATRIP complex is then activated by recruiting additional factors from the 9-1-1 complex (RAD9, RAD1, and HUS1), which subsequently recruits the TOPBP1 protein and represents a key step in the activation of the downstream phosphorylation cascade leading to cell cycle arrest. The main target of ATR kinase is CHK1, which, when phosphorylated, targets cdc25 and Wee1, leading to inhibition of cyclin-dependent kinase activity and cell cycle arrest at S phase or G2 / M phase.

[0004] ATR has been identified as an important cancer target because it is crucial for cell division. ATR-deficient mice are embryonic lethal; however, conditionally knocked-out ATR adult mice are viable and have an impact on rapidly proliferating tissues and stem cell populations. Embryonic stem cells in ATR-deficient mice only double 1-2 times before dying, indicating that ATR is necessary to maintain cell division. Interestingly, mice carrying hypomorphic ATR mutations (which reduce ATR expression to 10% of normal levels) show reduced H-rasG12D-induced tumor growth with minimal impact on proliferating normal cells (e.g., bone marrow or intestinal epithelial cells). Cancer cells with high levels of replication stress due to oncogenic mutations, dysfunctional G1 / S checkpoint regulation (e.g., p53 loss of function), defects in other DNA repair pathways (e.g., ATM), or those affected by DNA-damaging agents (e.g., radiotherapy or chemotherapy) are therefore more dependent on ATR for DNA repair and survival. In summary, these results highlight the fundamental principle of the selective sensitivity of proliferating tumor cells to ATR inhibition and the potential of a therapeutic window on healthy proliferating cells.

[0005] New anti-cancer therapies are needed, especially those based on ATR inhibitors. Summary of the Invention

[0006] In one aspect, the present invention provides a compound of formula (I):

[0007]

[0008] or a pharmaceutically acceptable salt thereof,

[0009] in

[0010] It is a double bond, and each Y is independently either N or CR. 4 ;or It is a single bond, and each Y is independently NR. Y carbonyl or C(R) Y )2; where each R Y Independently H or optionally substituted C 1-6 alkyl;

[0011] R 1 C is an optional replacement 1-6 Alkyl or H;

[0012] R 2 C is an optional replacement 2-9 Heterocyclic groups, optionally substituted C 1-6 Alkyl, optionally substituted C 3-8 cycloalkyl, optionally substituted C 2-9 Heterocyclic C1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 heteroaryl, optionally substituted C 1-9 heteroaryl C 1-6 Alkyl, halogen, –N(R) 5 2. –OR 5 –CON(R) 6 )2、–SO2N(R 6 )2、–SO2R 5A Or –Q–R 5B ;

[0013] R 3 C is an optional replacement 1-9 heteroaryl or optionally substituted C 1-9 heteroaryl C 1-6 alkyl;

[0014] Each R 4 Independently hydrogen, halogen, or optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 alkenyl or optionally substituted C 2-6 alkynyl group;

[0015] Each R 5 Independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 heteroaryl or –SO2R 5A ; or two Rs 5 Together with the atoms it is attached to, they combine to form optionally substituted C atoms. 2-9 Heterocyclic groups;

[0016] Each R 5A C can be substituted independently. 1-6 Alkyl, optionally substituted C 3-8 cycloalkyl or optionally substituted C 6-10 aryl;

[0017] R 5B Hydroxyl group, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 heteroaryl, –N(R) 5 )2、–CON(R 6 )2、–SO2N(R 6 )2、–SO2R 5A Or optionally substituted alkoxy groups;

[0018] Each R 6 Independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkoxyalkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 3-8 cycloalkyl or optionally substituted C 1-9 heteroaryl; or two R 6 Together with the atoms it is attached to, they combine to form optionally substituted C atoms. 2-9 Heterocyclic groups;

[0019] Q is an optional substitution for C. 2-9 Heterocyclic groups, optionally substituted C 3-8 Cycloalkylene, optionally substituted C 1-9 heteroaryl or optionally substituted C 6-10 Alpha-aryl; and

[0020] X is hydrogen or halogen.

[0021] In some embodiments, It is a double bond. In some implementations, It is a single key.

[0022] In some embodiments, the compound is a compound of formula (II):

[0023]

[0024] or a pharmaceutically acceptable salt thereof,

[0025] in

[0026] Each Y is independently either N or CR 4 ; and the remaining variables are as described for equation (I).

[0027] In some embodiments, in compounds of formula (I) or (II):

[0028] Each Y is independently either N or CR 4 ;

[0029] R 1 H or C with optional substitution 1-6 alkyl;

[0030] R 2 C is an optional replacement 1-6 Alkyl, optionally substituted C 3-8 cycloalkyl, optionally substituted C 2-9 Heterocyclic groups, optionally substituted C 6-10 aryl, optionally substituted C1-9 Heteroaryl, optionally substituted C 1-9 heteroaryl C 1-6 Alkyl, –N(R) 5 )2、–CON(R 6 )2、–SO2N(R 6 )2 or –SO2R 5A ;

[0031] R 3 C is an optional replacement 1-9 Mixed aromatics;

[0032] Each R 4 Independently hydrogen or optionally substituted C 1-6 alkyl;

[0033] Each R 5 Independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 heteroaryl or –SO2R 5A ; or two Rs 5 Together with the atoms it is attached to, they combine to form optionally substituted C atoms. 2-9 Heterocyclic groups;

[0034] Each R 5A C can be substituted independently. 1-6 Alkyl or optionally substituted C 3-8 cycloalkyl; and

[0035] Each R 6 Independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 aryl or optionally substituted C 1-9 heteroaryl; or two R 6 Together with the atoms it is attached to, they combine to form optionally substituted C atoms. 2-9 Heterocyclic group.

[0036] In some embodiments, the compound is a compound of formula (Ia):

[0037]

[0038] Or its pharmaceutically acceptable salt, wherein all variables are as described herein.

[0039] In some embodiments, the compound is a compound of formula (Ib):

[0040]

[0041]

[0042] Or its pharmaceutically acceptable salt, wherein all variables are as described herein.

[0043] In some embodiments, the compound is a compound of formula (IA):

[0044]

[0045] Or its pharmaceutically acceptable salt, wherein all variables are as described herein.

[0046] In some embodiments, the compound is a compound of formula (IA-a):

[0047]

[0048] Or its pharmaceutically acceptable salt, wherein all variables are as described herein.

[0049] In some implementations, the compound is a compound of formula (IB):

[0050]

[0051] Or its pharmaceutically acceptable salt, wherein all variables are as described herein.

[0052] In some embodiments, the compound is a compound of formula (IB-a):

[0053]

[0054] Or its pharmaceutically acceptable salt, wherein all variables are as described herein.

[0055] In some embodiments, the compound is a compound of formula (IC):

[0056]

[0057] Or its pharmaceutically acceptable salt, wherein all variables are as described herein.

[0058] In some embodiments, the compound is a compound of formula (IC-a):

[0059]

[0060] Or its pharmaceutically acceptable salt, wherein all variables are as described herein.

[0061] In some implementations, the compound is a compound of formula (ID):

[0062]

[0063] Or its pharmaceutically acceptable salt, wherein all variables are as described herein.

[0064] In some embodiments, the compound is a compound of formula (ID-a):

[0065]

[0066]

[0067] Or its pharmaceutically acceptable salt, wherein all variables are as described herein.

[0068] In some implementations of any of formulas (I), (II), (IA), (IA-a), (IB), (IB-a), (IC), (IC-a), (ID), and (ID-a), R 1 It is a methyl group.

[0069] In some implementations of any of formulas (I), (II), (IA), (IA-a), (IB), (IB-a), (IC), (IC-a), (ID), and (ID-a), R 2 C is an optional replacement 1-6 Alkyl, optionally substituted C 3-8 cycloalkyl, optionally substituted C 2-9 Heterocyclic groups, optionally substituted C 6-10 aryl, optionally substituted C 1-9 Heteroaryl, optionally substituted C 1-9 heteroaryl C 1-6 Alkyl, –N(R) 5 )2、–CON(R 6 )2、–SO2N(R 6 )2 or –SO2R 5A .

[0070] In some implementations of any of formulas (I), (II), (IA), (IA-a), (IB), (IB-a), (IC), (IC-a), (ID), and (ID-a), R 2 C is an optional replacement 3-8 Cycloalkyl. In some embodiments, R 2 Group (A) is:

[0071]

[0072] in

[0073] n is 0, 1, 2, or 3; and

[0074] R 7 Hydrogen, alkylsulfonyl, cyano, –CON(R) A )2、-SON(R A 2. Optional substitution of C 1-9 Heteroaryl, hydroxyl, or alkoxy, wherein each R A Independently H or alkyl; or two R A Together with the atoms it is attached to, they combine to form C. 2-9 Heterocyclic group.

[0075] In some implementations of any of formulas (I), (II), (IA), (IA-a), (IB), (IB-a), (IC), (IC-a), (ID), and (ID-a), R 2 For formula (B) group:

[0076]

[0077] where R 7 Hydrogen, alkylsulfonyl, cyano, –CON(R) A )2、-SON(R A 2. Optional substitution of C 1-9 Heteroaryl, hydroxyl, or alkoxy, wherein each R A Independently H or alkyl; or two R A Together with the atoms it is attached to, they combine to form C. 2-9 Heterocyclic group.

[0078] In some implementations of any of formulas (I), (II), (IA), (IA-a), (IB), (IB-a), (IC), (IC-a), (ID), and (ID-a), R 2 For optional substitution of non-aromatic C 2-9 Heterocyclic group. In some embodiments of any one of formulas (I), (II), (IA), (IA-a), (IB), (IB-a), (IC), (IC-a), (ID), and (ID-a), R 2 For optional replacement of non-aromatic bridges C 2-9 Heterocyclic group. In some embodiments of any one of formulas (I), (II), (IA), (IA-a), (IB), (IB-a), (IC), (IC-a), (ID), and (ID-a), R 2 For optional substitution of non-aromatic spirochetes C 2-9 Heterocyclic group.

[0079] In some implementations of any of formulas (I), (II), (IA), (IA-a), (IB), (IB-a), (IC), (IC-a), (ID), and (ID-a), R 2 For –Q–R 5B In some embodiments of any of formulas (I), (II), (IA), (IA-a), (IB), (IB-a), (IC), (IC-a), (ID), and (ID-a), Q is an optionally substituted C. 2-9 Subheterocyclic group. In some embodiments of any one of formulas (I), (II), (IA), (IA-a), (IB), (IB-a), (IC), (IC-a), (ID), and (ID-a), R 5B It is a hydroxyl group.

[0080] In some implementations of any of formulas (I), (II), (IA), (IA-a), (IB), (IB-a), (IC), (IC-a), (ID), and (ID-a), R 2 for:

[0081]

[0082] –I、–SO2Me、 –SO2Ph、 –OMe、 –OCH2CF3、

[0083]

[0084] In some implementations of any of formulas (I), (II), (IA), (IA-a), (IB), (IB-a), (IC), (IC-a), (ID), and (ID-a), R 3 A monocyclic C with optional substitution containing at least one nitrogen atom 1-9 Heteroaryl. In some embodiments of any one of formulas (I), (II), (IA), (IA-a), (IB), (IB-a), (IC), (IC-a), (ID), and (ID-a), R 3 A monocyclic C with optional substitution of two nitrogen atoms 1-9 Heteroaryl. In some embodiments of any one of formulas (I), (II), (IA), (IA-a), (IB), (IB-a), (IC), (IC-a), (ID), and (ID-a), R 3 For formula (C) group:

[0085]

[0086] Where A is an optionally substituted monocyclic C 1-9 Mixed aromatic rings.

[0087] In some implementations of any of formulas (I), (II), (IA), (IA-a), (IB), (IB-a), (IC), (IC-a), (ID), and (ID-a), R 3 For formula (C1) group:

[0088]

[0089] where R 8 C is hydrogen, halogen, or optionally substituted. 1-6 alkyl.

[0090] In some embodiments of any one of formulas (I), (II), (IA), (IA-a), (IB), (IB-a), (IC), (IC-a), (ID), and (ID-a), A is an optionally substituted monocyclic C containing two nitrogen atoms. 1-9 Mixed aromatic rings.

[0091] In some implementations of any of formulas (I), (II), (IA), (IA-a), (IB), (IB-a), (IC), (IC-a), (ID), and (ID-a), R 3 for:

[0092]

[0093]

[0094] In some implementations of any of formulas (I), (II), (IA), (IA-a), (IB), (IB-a), (IC), (IC-a), (ID), and (ID-a), R 3 for:

[0095]

[0096] In some implementations of any of formulas (I), (II), (IA), (IA-a), (IB), (IB-a), (IC), (IC-a), (ID), and (ID-a), R 4 For hydrogen.

[0097] In some embodiments of any one of formulas (I), (II), (IA), (IA-a), (IB), (IB-a), (IC), (IC-a), (ID), and (ID-a), X is hydrogen.

[0098] In some embodiments, the compounds are selected from the group consisting of compounds 1-152 (e.g., compounds 1-140) and their pharmaceutically acceptable salts (e.g., compounds selected from the group consisting of: 1, 2, 3, 4, 5, 6, 7, 8, 43, 45, 47, 48, 49, 52, 53, 55, 57, 58, 59, 61, 62, 63, 73, 74, 77, 80, 81, 82, 84, 86, 87, 92, 93, 94, 95, 97, 98, 99, 100, 101, 102, 1 03, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 118, 119, 120, 121, 122, 123, 125, 126, 127, 128, 129, 130, 131, 132, 133, 135, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 150, 151 and their pharmaceutically acceptable salts).

[0099] In another aspect, the present invention provides a pharmaceutical composition comprising the compound of the present invention and a pharmaceutically acceptable excipient. In some embodiments, the compound of the present invention is enriched with the isotope deuterium.

[0100] In another aspect, the present invention provides a method for inhibiting ATR kinase in cells by contacting cells expressing ATR kinase with the compounds of the present invention. In some embodiments, the cells are in vitro. In some embodiments, the cells are in a subject.

[0101] In another aspect, the present invention provides a method for treating a subject in need, the method comprising administering to the subject an effective amount of a compound of the present invention or a pharmaceutical composition of the present invention.

[0102] In some implementations, the subject is suffering from a disease or condition characterized by excessive cell proliferation (e.g., cancer or precancerous or malignant lesions) and requires treatment. In some implementations, the cancer is epithelial carcinoma, sarcoma, adenocarcinoma, leukemia, or melanoma.

[0103] In some implementations, the cancer is selected from the group consisting of: thyroid brain-like epithelial carcinoma, familial thyroid brain-like epithelial carcinoma, acinar epithelial carcinoma, acinar epithelial carcinoma, adenocystic epithelial carcinoma, adenoid cystic epithelial carcinoma, adenomatous epithelial carcinoma, adrenocortical epithelial carcinoma, alveolar epithelial carcinoma, alveolar cell epithelial carcinoma, basal cell epithelial carcinoma, basal cell carcinoma, basaloid epithelial carcinoma, basal squamous cell carcinoma, bronchioloalveolar epithelial carcinoma, bronchiolar epithelial carcinoma, bronchial epithelial carcinoma, cerebriform carcinoma, cholangiocarcinoma, colloid epithelial carcinoma, comedo carcinoma, corpus carcinoma, cribriform epithelial carcinoma, armored epithelial carcinoma, cutaneous epithelial carcinoma, columnar epithelial carcinoma, columnar cell epithelial carcinoma, ductal epithelial carcinoma, and sclerothelial carcinoma. Durum, embryonic epithelial carcinoma, encephaloid carcinoma, epidermoid carcinoma, adenoid carcinoma, exophytic epithelial carcinoma, ulcerative epithelial carcinoma, fibrous epithelial carcinoma, gelatiniforni carcinoma, gelatinous carcinoma, giant cell carcinoma, gigantocellular carcinoma, adenocarcinoma, granular cell carcinoma, pilostromal carcinoma, hematoid carcinoma, hepatocellular carcinoma, Hurthle cell carcinoma, hyaline carcinoma, hypernephroid Carcinoma, embryonic carcinoma, epithelial carcinoma in situ, epithelial intraepithelial carcinoma, intraepithelial carcinoma, Krompecher's carcinoma, Kulchitzky-cell carcinoma, large cell carcinoma, lenticular carcinoma, lenticular carcinoma, lipoma carcinoma, lymphoepithelial carcinoma, medullary carcinoma, medullary carcinoma, melanotic carcinoma.Carcinoma, soft carcinoma, mucinous carcinoma, muciparum carcinoma, mucocellular carcinoma, mucoepidermoid carcinoma, mucocellular carcinoma, mucous carcinoma, myxomatodes carcinoma, nasopharyngeal carcinoma, oat cell carcinoma, ossifying carcinoma, osteoid carcinoma, papillary carcinoma, periportal carcinoma, invasive preepithelial carcinoma, prickle cell carcinoma, and pultaceous carcinoma. Renal cell carcinoma, reserve cell carcinoma, sarcomatodes carcinoma, Schneiderian carcinoma, scirrhous carcinoma, scroti carcinoma, signet-ring cell carcinoma, simplex carcinoma, small cell carcinoma, solanoid carcinoma, spheroidal cell carcinoma, spindle cell carcinoma, spongy carcinoma, squamous carcinoma, string carcinoma, telangiectaticum carcinoma, and angiotensinus carcinoma. telangiectodes, transitional cell carcinoma, nodular carcinomaTuberculous carcinoma (TBC), tuberous carcinoma, verrucous carcinoma, and villosum carcinoma.

[0104] In some implementations, the cancer is a sarcoma selected from the group consisting of: chondrosarcoma, fibrosarcoma, lymphosarcoma, melanosarcoma, myxosarcoma, osteosarcoma, Abernethy's sarcoma, liposarcoma, liposarcoma, alveolar soft tissue sarcoma, ameloblastic sarcoma, botryoid sarcoma, chloromasarcoma, choriocarcinoma, embryonal sarcoma, Wilms' tumor sarcoma, endometrial sarcoma, stromal sarcoma, Ewing's sarcoma, fascial sarcoma, fibroblastic sarcoma, giant cell sarcoma, granulocytic sarcoma, Hodgkin's sarcoma, and idiopathic multiple pigmented hemorrhagic sarcoma. Sarcoma, B-cell immunoblastic sarcoma, T-cell immunoblastic sarcoma, Jensen's sarcoma, Kaposi's sarcoma, Kupffer cell sarcoma, angiosarcoma, leukemic sarcoma, malignant mesenchymoma sarcoma, periosteal sarcoma, reticulocytic sarcoma, Rous sarcoma, serocystic sarcoma, synovial sarcoma, and telangiectaltic sarcoma.

[0105] In some implementation schemes, cancer is selected from the group consisting of: non-lymphocytic leukemia, chronic lymphocytic leukemia, acute granulocytic leukemia, chronic myeloid leukemia, acute promyelocytic leukemia, adult T-cell leukemia, non-leukemic leukemia, leukemic leukemia, basophageal leukemia, blastic leukemia, bovine leukemia, chronic myeloid leukemia, cutaneous leukemia, embryonal leukemia, eosinophilic leukemia, Gross' leukemia, hairy cell leukemia, hemoblastic leukemia, and hemocytoblastic leukemia. Leukemia, histiocytic leukemia, stem cell leukemia, acute monocytic leukemia, leukopenic leukemia, lymphatic leukemia, lymphoblastic leukemia, lymphocyte leukemia, lymphogenous leukemia, lymphoid leukemia, lymphosarcoma cell leukemia, mast cell leukemia, megakaryocytic leukemia, micromyeloblastic leukemia, monocytic leukemia, myeloblastic leukemia, myelocyticle leukemia, myeloid granulocytic leukemia, myelomonocytic leukemia, Naegeli Leukemia, plasma cell leukemia, multiple myeloma, plasma cell leukemia, promyelocytic leukemia, Rieder cell leukemia, Schilling's leukemia, stem cell leukemia, subleukemic leukemia, and undifferentiated cell leukemia.

[0106] In some implementations, the cancer is melanoma selected from the group consisting of: acral-lentiginous melanoma, amelanoma, benign juvenile melanoma, Cloudman's melanoma, S91 melanoma, Harding-Passey melanoma, juvenile melanoma, lentigo maligna melanoma, malignant melanoma, nodular melanoma, subungual melanoma, and superficial diffuse melanoma.

[0107] In some implementation schemes, the cancer is prostate cancer, thyroid cancer, endocrine system cancer, brain cancer, breast cancer, cervical cancer, colon cancer, head and neck cancer, liver cancer, kidney cancer, lung cancer, non-small cell lung cancer, melanoma, mesothelioma, ovarian cancer, sarcoma, stomach cancer, uterine cancer, medulloblastoma, ampullary cancer, colorectal cancer, or pancreatic cancer.

[0108] In some implementation schemes, cancer includes Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, neuroblastoma, glioma, glioblastoma multiforme, ovarian cancer, rhabdomyosarcoma, essential thrombocytosis, essential macroglobulinemia, primary brain tumor, cancer, malignant pancreatic insulinoma, malignant carcinoid tumor, bladder cancer, precancerous skin lesions, testicular cancer, lymphoma, thyroid cancer, esophageal cancer, urogenital tract cancer, malignant hypercalcemia, endometrial cancer, adrenocortical cancer, endocrine and exocrine pancreatic growths, medullary thyroid carcinoma, thyroid brain-like epithelial carcinoma, melanoma, colorectal cancer, papillary thyroid carcinoma, hepatocellular carcinoma, or prostate cancer.

[0109] In some implementations, the subject is suffering from pre-malignant conditions and requires treatment.

[0110] The invention is also described through the following list of items.

[0111] 1. A compound of formula (I):

[0112]

[0113] or a pharmaceutically acceptable salt thereof,

[0114] in

[0115] It is a double bond, and each Y is independently either N or CR. 4 ;or It is a single bond, and each Y is independently NR. Ycarbonyl or C(R) Y )2; where each R Y Independently H or optionally substituted C 1-6 alkyl;

[0116] R 1 C is an optional replacement 1-6 Alkyl or H;

[0117] R 2 C is an optional replacement 2-9 Heterocyclic groups, optionally substituted C 1-6 Alkyl, optionally substituted C 3-8 cycloalkyl, optionally substituted C 2-9 Heterocyclic C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 Heteroaryl, optionally substituted C 1-9 heteroaryl C 1-6 Alkyl, halogen, –N(R) 5 2. –OR 5 –CON(R) 6 )2、–SO2N(R 6 )2、–SO2R 5A Or –Q–R 5B ;

[0118] R 3 C is an optional replacement 1-9 heteroaryl or optionally substituted C 1-9 heteroaryl C 1-6 alkyl;

[0119] Each R 4 Independently hydrogen, halogen, or optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 alkenyl or optionally substituted C 2-6 alkynyl group;

[0120] Each R 5 Independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 heteroaryl or –SO2R 5A ; or two Rs 5 Together with the atoms it is attached to, they combine to form optionally substituted C atoms. 2-9 Heterocyclic groups;

[0121] Each R 5A C can be substituted independently. 1-6Alkyl, optionally substituted C 3-8 cycloalkyl or optionally substituted C 6-10 aryl;

[0122] R 5B Hydroxyl group, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 heteroaryl, –N(R) 5 )2、–CON(R 6 )2、–SO2N(R 6 )2、–SO2R 5A Or optionally substituted alkoxy groups;

[0123] Each R 6 Independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkoxyalkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 3-8 cycloalkyl or optionally substituted C 1-9 heteroaryl; or two R 6 Together with the atoms it is attached to, they combine to form optionally substituted C atoms. 2-9 Heterocyclic groups;

[0124] Q is an optional substitution for C. 2-9 Heterocyclic groups, optionally substituted C 3-8 Cycloalkylene, optionally substituted C 1-9 heteroaryl or optionally substituted C 6-10 Alpha-aryl; and

[0125] X is hydrogen or halogen.

[0126] 2. The compound as described in Project 1, wherein It is a double bond.

[0127] 3. The compound as described in Project 1, wherein It is a single key.

[0128] 4. The compound as described in Project 1, wherein the compound is a compound of formula (II):

[0129]

[0130] Or its pharmaceutically acceptable salt.

[0131] in

[0132] Each Y is independently either N or CR 4 ;

[0133] R 1 C is an optional replacement 1-6 Alkyl or H;

[0134] R 2 C is an optional replacement 2-9 Heterocyclic groups, optionally substituted C 1-6 Alkyl, optionally substituted C 3-8 cycloalkyl, optionally substituted C 2-9 Heterocyclic C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 Heteroaryl, optionally substituted C 1-9 heteroaryl C 1-6 Alkyl, halogen, –N(R) 5 2. –OR 5 –CON(R) 6 )2、–SO2N(R 6 )2、–SO2R 5A Or –Q–R 5B ;

[0135] R 3 C is an optional replacement 1-9 heteroaryl or optionally substituted C 1-9 heteroaryl C 1-6 alkyl;

[0136] Each R 4 Independently hydrogen, halogen, or optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 alkenyl or optionally substituted C 2-6 alkynyl group;

[0137] Each R 5 Independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 heteroaryl or –SO2R 5A ; or two Rs 5 Together with the atoms it is attached to, they combine to form optionally substituted C atoms. 2-9 Heterocyclic groups;

[0138] Each R 5A C can be substituted independently. 1-6 Alkyl, optionally substituted C 3-8 cycloalkyl or optionally substituted C 6-10 aryl;

[0139] R 5B Hydroxyl group, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 heteroaryl, –N(R) 5 )2、–CON(R 6 )2、–SO2N(R 6 )2、–SO2R 5A Or optionally substituted alkoxy groups;

[0140] Each R 6 Independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkoxyalkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 3-8 cycloalkyl or optionally substituted C 1-9 heteroaryl; or two R 6 Together with the atoms it is attached to, they combine to form optionally substituted C atoms. 2-9 Heterocyclic groups;

[0141] Q is an optional substitution for C. 2-9 Heterocyclic groups, optionally substituted C 3-8 Cycloalkylene, optionally substituted C 1-9 heteroaryl or optionally substituted C 6-10 Alpha-aryl; and

[0142] X is hydrogen or halogen.

[0143] 5. The compound as described in Project 1, wherein the compound is a compound of formula (Ia):

[0144]

[0145] Or its pharmaceutically acceptable salt.

[0146] 6. The compound as described in Project 1, wherein the compound is a compound of formula (IA):

[0147]

[0148] Or its pharmaceutically acceptable salt.

[0149] 7. The compound as described in item 6, wherein the compound is a compound of formula (IA-a):

[0150]

[0151] Or its pharmaceutically acceptable salt.

[0152] 8. The compound as described in Item 1, wherein the compound is a compound of formula (IB):

[0153]

[0154] Or its pharmaceutically acceptable salt.

[0155] 9. The compound as described in item 8, wherein the compound is a compound of formula (IB-a):

[0156]

[0157] Or its pharmaceutically acceptable salt.

[0158] 10. The compound as described in Project 1, wherein the compound is a compound of formula (IC):

[0159]

[0160] Or its pharmaceutically acceptable salt.

[0161] 11. The compound as described in item 10, wherein the compound is a compound of formula (IC-a):

[0162]

[0163] Or its pharmaceutically acceptable salt.

[0164] 12. The compound of any one of items 1 to 11, wherein R 1 It is a methyl group.

[0165] 13. The compound of any one of items 1 to 12, wherein R 2 C is an optional replacement 1-6 Alkyl, optionally substituted C 3-8 cycloalkyl, optionally substituted C 2-9 Heterocyclic groups, optionally substituted C 6-10 aryl, optionally substituted C 1-9 Heteroaryl, optionally substituted C 1-9 heteroaryl C 1-6 Alkyl, –N(R) 5 )2、–CON(R 6 )2、–SO2N(R 6 )2 or –SO2R 5A .

[0166] 14. The compound as described in any one of items 1 to 13, wherein each R 5A C can be substituted independently. 1-6Alkyl or optionally substituted C 3-8 Cycloalkyl.

[0167] 15. The compound as described in any one of items 1 to 13, wherein each R 6 Independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 aryl or optionally substituted C 1-9 heteroaryl; or two R 6 Together with the atoms it is attached to, they combine to form optionally substituted C atoms. 2-9 Heterocyclic group.

[0168] 16. The compound of any one of items 1 to 15, wherein R 2 C is an optional replacement 3-8 Cycloalkyl.

[0169] 17. The compound as described in item 16, wherein R 2 C 3-8 Cycloalkyl groups, optionally prefixed with alkylsulfonyl, cyano, or –CON(R) groups. A 2. Hydroxyl or alkoxy substitution, wherein each R A Independently H or alkyl; or R A Together with the atoms it is attached to, they combine to form C. 2-9 Heterocyclic group.

[0170] 18. The compound as described in item 16, wherein R 2 Group (A) is:

[0171]

[0172] in

[0173] n is 0, 1, 2, or 3; and

[0174] R 7 Hydrogen, alkylsulfonyl, cyano, –CON(R) A )2、-SON(R A 2. Optional substitution of C 1-9 Heteroaryl, hydroxyl, or alkoxy, wherein each R A Independently H or alkyl; or two R A Together with the atoms it is attached to, they combine to form C. 2-9 Heterocyclic group.

[0175] 19. The compound of any one of items 1 to 15, wherein R 2 For formula (B) group:

[0176]

[0177] Where R 7 Hydrogen, alkylsulfonyl, cyano, –CON(R) A )2、-SON(R A 2. Optional substitution of C 1-9 Heteroaryl, hydroxyl, or alkoxy, wherein each R A Independently H or alkyl; or two R A Together with the atoms it is attached to, they combine to form C. 2-9 Heterocyclic group.

[0178] 20. The compound as described in item 18 or 19, wherein R 7 It is an alkylsulfonyl, cyano, or –CON(R) group. A )2.

[0179] 21. The compound of any one of items 1 to 12, wherein R 2 C is an optional replacement 1-6 alkyl.

[0180] 22. The compound as described in item 21, wherein R 2 For optional replacement of uncle C 3-6 alkyl.

[0181] 23. The compound of any one of items 1 to 15, wherein R 2 For optional substitution of non-aromatic C 2-9 Heterocyclic group.

[0182] 24. The compound as described in item 23, wherein R 2 For optional replacement of non-aromatic bridges C 2-9 Heterocyclic group.

[0183] 25. The compound as described in item 23, wherein R 2 For optional substitution of non-aromatic spirochetes C 2-9 Heterocyclic group.

[0184] 26. The compound of any one of items 1 to 15, wherein R 2 C is an optional replacement 3-8 Cycloalkyl.

[0185] 27. The compound as described in item 26, wherein R 2 For optional replacement of screw C 3-8 Cycloalkyl.

[0186] 28. The compound of any one of items 1 to 12, wherein R 2 For –Q–R 5B .

[0187] 29. The compound as described in item 28, wherein Q is an optionally substituted C. 1-9 Hybrid aryl.

[0188] 30. The compound as described in item 28, wherein Q is an optionally substituted C. 3-8 Cycloalkylene.

[0189] 31. The compound as described in Item 28, wherein Q is an optionally substituted C. 2-9 Sub-heterocyclic group.

[0190] 32. The compound as described in item 28, wherein Q is an optionally substituted C. 6-10 Alpha-aryl.

[0191] 33. The compound of any one of items 28 to 32, wherein R 5B C is an optional replacement 1-6 alkyl.

[0192] 34. The compound of any one of items 28 to 32, wherein R 5B It is a hydroxyl group.

[0193] 35. The compound of any one of items 28 to 32, wherein R 5B C is an optional replacement 6-10 Aryl.

[0194] 36. The compound of any one of items 28 to 32, wherein R 5B C is an optional replacement 1-9 Mixed aromatic compounds.

[0195] 37. The compound of any one of items 28 to 32, wherein R 5B For –N(R) 5 )2.

[0196] 38. The compound as described in item 37, wherein each R 5 For hydrogen.

[0197] 39. The compound of any one of items 28 to 32, wherein R 5B The alkoxy group is optionally substituted.

[0198] 38. The compound of any one of items 28 to 32, wherein R 5B -SO2N(R) 6 )2.

[0199] 39. The compound as described in item 38, wherein each R 6 For hydrogen.

[0200] 40. The compound of any one of items 28 to 32, wherein R 5B For –SO2R 5A .

[0201] 41. The compound as described in item 40, wherein R 5A C is an optional replacement 1-6 alkyl.

[0202] 42. The compound of any one of items 1 to 15, wherein R 2 for:

[0203]

[0204] –I、–SO2Me、 –SO2Ph、 –OMe、 –OCH2CF3、

[0205]

[0206] 43. The compound as described in item 42, wherein R 2 for:

[0207]

[0208] –I、–SO2Me、 –SO2Ph、 –OMe、 –OCH2CF3、

[0209]

[0210] 44. The compound as described in item 42, wherein R 2 for:

[0211]

[0212]

[0213] 45. The compound as described in item 42, wherein R 2 for:

[0214] –OCH2CF3、

[0215]

[0216] 46. ​​The compound as described in item 42, wherein R 2 for:

[0217]

[0218] 47. The compound as described in item 42, wherein R 2 for:

[0219]

[0220] 48. The compound as described in item 42, wherein R 2 for:

[0221]

[0222] 49. The compound as described in item 42, wherein R 2 for:

[0223]

[0224] 50. The compound as described in item 42, wherein R 2 for:

[0225]

[0226] 51. The compound as described in item 42, wherein R 2 for:

[0227]

[0228] 52. The compound as described in item 42, wherein R 2 for:

[0229]

[0230] 53. The compound as described in item 42, wherein R 2 for:

[0231]

[0232] 54. The compound as described in item 42, wherein R 2 for:

[0233]

[0234] 55. The compound as described in item 42, wherein R 2 for:

[0235]

[0236] 56. The compound as described in item 42, wherein R 2 for:

[0237]

[0238] 57. The compound as described in item 42, wherein R 2 for:

[0239]

[0240] 58. The compound as described in item 42, wherein R 2 for:

[0241]

[0242] 59. The compound as described in item 42, wherein R 2 for:

[0243]

[0244] 60. The compound as described in item 42, wherein R 2 for:

[0245]

[0246] 61. The compound as described in item 42, wherein R 2 for:

[0247]

[0248] 62. The compound as described in item 61, wherein R 2 for:

[0249]

[0250] 63. The compound as described in item 42, wherein R 2 for:

[0251]

[0252] 64. The compound as described in item 42, wherein R 2 for:

[0253]

[0254] 65. The compound as described in item 42, wherein R 2 for:

[0255]

[0256] 66. The compound as described in item 42, wherein R 2 for:

[0257]

[0258] 67. The compound as described in item 42, wherein R 2 for:

[0259]

[0260] 68. The compound as described in item 42, wherein R 2 for:

[0261]

[0262] 69. The compound as described in item 42, wherein R 2 for:

[0263]

[0264] 70. The compound of any one of items 1 to 69, wherein R 3 A monocyclic C with optional substitution containing at least one nitrogen atom 1-9 Mixed aromatic compounds.

[0265] 71. The compound as described in item 70, wherein R 3 A monocyclic C with optional substitution of two nitrogen atoms 1-9 Mixed aromatic compounds.

[0266] 72. The compound as described in item 70, wherein R 3 For formula (C) group:

[0267]

[0268] Where A is an optionally substituted monocyclic C 1-9 Mixed aromatic rings.

[0269] 73. The compound as described in item 70, wherein R 3 For formula (C1) group:

[0270]

[0271] Where R 8 C is hydrogen, halogen, or optionally substituted. 1-6 alkyl.

[0272] 74. The compound as described in item 73, wherein R 8 It is hydrogen or halogen.

[0273] 75. The compound of any one of items 72 to 74, wherein A is an optionally substituted monocyclic C containing two nitrogen atoms. 1-9 Mixed aromatic rings.

[0274] 76. The compound of any one of items 1 to 75, wherein R 3 for:

[0275]

[0276] 77. The compound as described in item 76, wherein R 3 for:

[0277]

[0278] 78. The compound as described in item 76, wherein R 3 for:

[0279]

[0280] 79. The compound as described in item 76, wherein R 3 for:

[0281]

[0282] 80. The compound as described in item 76, wherein R 3 for:

[0283]

[0284] 81. The compound of any one of items 1 to 80, wherein R 4 For hydrogen.

[0285] 82. The compound of any one of items 1 to 80, wherein R 4 It is a halogen.

[0286] 83. The compound of any one of items 1 to 80, wherein R 4 C is an optional replacement 2-6 Alkenyl.

[0287] 84. The compound of any one of items 1 to 80, wherein X is hydrogen.

[0288] 85. A compound selected from the group consisting of compounds 1-152 and their pharmaceutically acceptable salts.

[0289] 87. The compound as described in item 85, wherein the compound is selected from the group consisting of:

[0290] Compounds 1, 2, 3, 4, 5, 6, 7, 8, 24, 43, 45, 47, 48, 49, 52, 53, 55, 57, 58, 59, 61, 62, 63, 73, 74, 77, 80, 81, 82, 84, 86, 87, 92, 93, 94, 95, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110 111, 112, 113, 114, 115, 116, 118, 119, 120, 121, 122, 123, 125, 126, 127, 128, 129, 130, 131, 132, 133, 135, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 150, 151 and their pharmaceutically acceptable salts.

[0291] 88. The compound of claim 85, wherein the compound is selected from the group consisting of compounds 6, 8, 43, 48, 92, 126, 128, 130, 131, 141, 142, 143, 145, 150 and their pharmaceutically acceptable salts.

[0292] 89. The compound as described in item 85, wherein the compound is selected from the group consisting of compounds 2, 4, 7, 47, 49, 63, 86 and their pharmaceutically acceptable salts.

[0293] 90. The compound of claim 85, wherein the compound is selected from the group consisting of: compounds 57, 62, 73, 74, 80, 81, 82, 84, 87, 93, 94, 95, 99, 100, 106, 107, 108, 109, 111, 112, 113, 114, 115, 116, 118, 119, 120, 121, 122, 123, 135, 137, 138, 144, 146, 147, 148, 151 and pharmaceutically acceptable salts thereof.

[0294] 91. The compound of claim 85, wherein the compound is selected from the group consisting of: compounds 57, 62, 87, 93, 94, 95, 99, 100, 106, 107, 108, 109, 111, 112, 113, 114, 115, 116, 118, 119, 120, 121, 122, 123, 135, 147, 148 and their pharmaceutically acceptable salts.

[0295] 92. The compound as described in item 85, wherein the compound is selected from the group consisting of compounds 61, 105, 107, 110, 112, 113 and their pharmaceutically acceptable salts.

[0296] 93. The compound as described in item 56, wherein the compound is selected from the group consisting of compounds 121, 122 and their pharmaceutically acceptable salts.

[0297] 94. The compound as described in item 85, wherein the compound is selected from the group consisting of compounds 125, 127, 129, 138, 139, 140, 144, 146, 151 and their pharmaceutically acceptable salts.

[0298] 95. The compound as described in item 85, wherein the compound is selected from the group consisting of compounds 58, 123 and their pharmaceutically acceptable salts.

[0299] 96. The compound of claim 85, wherein the compound is selected from the group consisting of compounds 1, 3, 5, 59, 77, 97, 98, 101, 102, 103, 104, 106, 114, 115, 132, 133 and their pharmaceutically acceptable salts.

[0300] 97. The compound as described in item 85, wherein the compound is selected from the group consisting of compounds 45, 52, 55 and their pharmaceutically acceptable salts.

[0301] 98. The compound as described in item 85, wherein the compound is compound 1 or a pharmaceutically acceptable salt thereof.

[0302] 99. The compound as described in item 85, wherein the compound is compound 2 or a pharmaceutically acceptable salt thereof.

[0303] 100. The compound as described in item 85, wherein the compound is compound 3 or a pharmaceutically acceptable salt thereof.

[0304] 101. The compound as described in item 85, wherein the compound is compound 4 or a pharmaceutically acceptable salt thereof.

[0305] 102. The compound as described in item 85, wherein the compound is compound 5 or a pharmaceutically acceptable salt thereof.

[0306] 103. The compound as described in item 85, wherein the compound is compound 6 or a pharmaceutically acceptable salt thereof.

[0307] 104. The compound as described in item 85, wherein the compound is compound 7 or a pharmaceutically acceptable salt thereof.

[0308] 105. The compound as described in item 85, wherein the compound is compound 8 or a pharmaceutically acceptable salt thereof.

[0309] 106. The compound as described in item 85, wherein the compound is compound 9 or a pharmaceutically acceptable salt thereof.

[0310] 107. The compound as described in item 85, wherein the compound is compound 86 or a pharmaceutically acceptable salt thereof.

[0311] 108. The compound as described in item 85, wherein the compound is compound 99 or a pharmaceutically acceptable salt thereof.

[0312] 109. The compound as described in item 85, wherein the compound is compound 100 or a pharmaceutically acceptable salt thereof.

[0313] 110. The compound as described in item 85, wherein the compound is compound 115 or a pharmaceutically acceptable salt thereof.

[0314] 111. The compound as described in item 85, wherein the compound is compound 120 or a pharmaceutically acceptable salt thereof.

[0315] 112. The compound as described in item 85, wherein the compound is compound 121 or a pharmaceutically acceptable salt thereof.

[0316] 113. The compound as described in item 85, wherein the compound is compound 125 or a pharmaceutically acceptable salt thereof.

[0317] 114. The compound as described in item 85, wherein the compound is compound 126 or a pharmaceutically acceptable salt thereof.

[0318] 115. The compound as described in item 85, wherein the compound is compound 138 or a pharmaceutically acceptable salt thereof.

[0319] 116. The compound as described in item 85, wherein the compound is compound 139 or a pharmaceutically acceptable salt thereof.

[0320] 117. The compound as described in item 85, wherein the compound is compound 140 or a pharmaceutically acceptable salt thereof.

[0321] 118. The compound as described in item 85, wherein the compound is compound 142 or a pharmaceutically acceptable salt thereof.

[0322] 119. The compound as described in item 85, wherein the compound is compound 144 or a pharmaceutically acceptable salt thereof.

[0323] 120. The compound as described in item 85, wherein the compound is compound 147 or a pharmaceutically acceptable salt thereof.

[0324] 121. The compound as described in item 85, wherein the compound is compound 148 or a pharmaceutically acceptable salt thereof.

[0325] 122. The compound as described in item 85, wherein the compound is compound 150 or a pharmaceutically acceptable salt thereof.

[0326] 123. The compound as described in item 85, wherein the compound is compound 151 or a pharmaceutically acceptable salt thereof.

[0327] 124. A pharmaceutical composition comprising a compound as described in any one of items 1 to 123 and a pharmaceutically acceptable excipient.

[0328] 125. The pharmaceutical composition of item 124, wherein the compound is enriched with the isotope deuterium.

[0329] 126. A method for inhibiting ATR kinase in cells expressing ATR kinase, the method comprising contacting the cells with a compound as described in any one of items 1 to 123.

[0330] 127. The method of claim 126, wherein the cells are in vitro.

[0331] 128. The method as described in item 126, wherein the cells are in the subject.

[0332] 129. A method of treating a subject in need, comprising administering to the subject a compound as described in any one of items 1 to 123 or a pharmaceutical composition as described in item 124 or 125.

[0333] 130. The method as described in item 128 or 129, wherein the subject is suffering from a disease or condition characterized by excessive cell proliferation and requires treatment.

[0334] 131. The method as described in item 130, wherein the disease or symptom is cancer.

[0335] 132. The method as described in item 131, wherein the cancer is a solid tumor.

[0336] 133. The method as described in item 131, wherein the cancer is epithelial carcinoma, sarcoma, adenocarcinoma, leukemia, or melanoma.

[0337] 134. The method as described in Item 131, wherein the cancer is an epithelial carcinoma selected from the group consisting of: thyroid brain-like epithelial carcinoma, familial thyroid brain-like epithelial carcinoma, acinar epithelial carcinoma, acinar epithelial carcinoma, adenoid cystic epithelial carcinoma, adenoid cystic epithelial carcinoma, adenomatous epithelial carcinoma, adrenocortical epithelial carcinoma, alveolar epithelial carcinoma, alveolar cell epithelial carcinoma, basal cell epithelial carcinoma, basal cell carcinoma, basaloid epithelial carcinoma, basal squamous cell carcinoma, bronchioloalveolar epithelial carcinoma, bronchiolar epithelial carcinoma, bronchiolar epithelial carcinoma, medullary epithelial carcinoma, cholangiocarcinoma, choriocarcinoma. Skin carcinoma, colloid carcinoma, comedo-like carcinoma, main carcinoma, cribriform carcinoma, armored carcinoma, cutaneous carcinoma, columnar carcinoma, columnar cell carcinoma, ductal carcinoma, sclerothelial carcinoma, embryonal carcinoma, cerebral carcinoma, epidermoid carcinoma, adenoid carcinoma, exophytic carcinoma, ulcerative carcinoma, fibrocarcinoma, colloid carcinoma, gelatinous carcinoma, giant cell carcinoma, giant cell carcinoma, adenocarcinoma, granular cell carcinoma, pilostromal carcinoma, polycythian carcinoma, hepatocellular carcinoma, Schulte's cell carcinoma, hyaline carcinoma, adrenal carcinoma Epithelial carcinoma, infantile embryonic epithelial carcinoma, epithelial carcinoma in situ, intraepithelial epithelial carcinoma, intraepithelial intraepithelial carcinoma, Klinefelter's cell carcinoma, Kurtschitzky cell carcinoma, large cell epithelial carcinoma, lenticular epithelial carcinoma, lenticular epithelial carcinoma, lipomatous epithelial carcinoma, lymphoepithelial carcinoma, medullary epithelial carcinoma, medullary epithelial carcinoma, melanocyte epithelial carcinoma, soft epithelial carcinoma, mucinous epithelial carcinoma, mucinous cell epithelial carcinoma, mucinous epithelial carcinoma, mucinous epithelial carcinoma, myxomatous epithelial carcinoma, nasopharyngeal epithelial carcinoma, oat cell epithelial carcinoma, ossifying epithelial carcinoma, osteoid epithelial carcinoma, papillary epithelial carcinoma, periportal epithelial carcinoma Skin carcinoma, invasive preepithelial carcinoma, acanthosis nigra, soft pasty carcinoma, renal cell carcinoma, reservoir cell carcinoma, sarcomatoid carcinoma, Schneider's carcinoma, sclerosing carcinoma, scrotal carcinoma, signet ring cell carcinoma, simple carcinoma, small cell carcinoma, potato-shaped carcinoma, spherical cell carcinoma, spindle cell carcinoma, cavernous carcinoma, squamous cell carcinoma, string carcinoma, capillary carcinoma, angiotensinus carcinoma, transitional cell carcinoma, nodular carcinoma, nodular epithelial carcinoma, verrucous carcinoma, and villous carcinoma.

[0338] 135. The method as described in Item 131, wherein the cancer is a sarcoma selected from the group consisting of: chondrosarcoma, fibrosarcoma, lymphosarcoma, melanosarcoma, myxosarcoma, osteosarcoma, Ebernathy's sarcoma, liposarcoma, liposarcoma, alveolar soft tissue sarcoma, ameloblastic sarcoma, botryoid sarcoma, chloroma sarcoma, choriocarcinoma, embryonal sarcoma, Williams' tumor sarcoma, endometrial sarcoma, stromal sarcoma, Ewing's sarcoma, and fascia. Membranous sarcoma, fibroblastic sarcoma, giant cell sarcoma, granulocyte sarcoma, Hodgkin's sarcoma, idiopathic multiple pigmented hemorrhagic sarcoma, B-cell immunoblastic sarcoma, T-cell immunoblastic sarcoma, Jensen's sarcoma, Kaposi's sarcoma, Kupffer cell sarcoma, angiosarcoma, leukemic sarcoma, malignant mesenchymal sarcoma, paraosteal sarcoma, reticulum cell sarcoma, Rouss's sarcoma, serous cystic sarcoma, synovial sarcoma, and capillary hemangiosarcoma.

[0339] 136. The method as described in Item 131, wherein the cancer is a leukemia selected from the group consisting of: non-lymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, acute promyelocytic leukemia, adult T-cell leukemia, non-leukemic leukemia, leukemic leukemia, basophilic leukemia, blastic leukemia, bovine leukemia, chronic myeloid leukemia, cutaneous leukemia, embryonic leukemia, eosinophilic leukemia, Gross's leukemia, hairy cell leukemia, hematopoietic leukemia, hematopoietic leukemia, histiocytic leukemia, stem cell leukemia, acute monocytic leukemia. Leukemia, leukopenic leukemia, lymphocytic leukemia, lymphoblastic leukemia, lymphocytic leukemia, lymphoid leukemia, lymphosarcoma cell leukemia, mast cell leukemia, megakaryocytic leukemia, micromyeloid leukemia, monocytic leukemia, myeloblastic leukemia, myeloid leukemia, myeloid granulocytic leukemia, granulocytic-monocytic leukemia, Neghly leukemia, plasma cell leukemia, multiple myeloma, plasma cell leukemia, promyelocytic leukemia, Riddle cell leukemia, Schilling leukemia, stem cell leukemia, subleukemic leukemia, and undifferentiated cell leukemia.

[0340] 137. The method as described in item 136, wherein the cancer is chronic lymphocytic leukemia.

[0341] 138. The method as described in item 131, wherein the cancer is lymphoma.

[0342] 139. The method as described in Item 138, wherein the lymphoma is non-Hodgkin's lymphoma, Hodgkin's disease, diffuse large B-cell lymphoma, follicular lymphoma, mucosa-associated lymphoid tissue (MALT) lymphoma, small cell lymphocytic lymphoma-chronic lymphocytic leukemia, mantle cell lymphoma, mediastinal (thymic) large B-cell lymphoma, lymphoplasmacytic lymphoma-Wald's macroglobulinemia, peripheral T-cell lymphoma (PTCL), angioimmunoblastic T-cell lymphoma (AITL) / follicular T-cell lymphoma (FTCL), anaplastic large cell lymphoma (ALCL), enteropathy-associated T-cell lymphoma (EATL), adult T-cell leukemia / lymphoma (ATLL), or extranodal NK / T-cell lymphoma (nasal type).

[0343] 140. The method of claim 139, wherein the lymphoma is mantle cell lymphoma.

[0344] 141. The method of claim 131, wherein the cancer is a melanoma selected from the group consisting of: acral lentigines melanoma, amelanoma, benign juvenile melanoma, Claudemann's melanoma, S91 melanoma, Harpa's II melanoma, juvenile melanoma, malignant lentigines melanoma, malignant melanoma, nodular melanoma, subungual melanoma, and superficial diffuse melanoma.

[0345] 142. The method as described in Item 131, wherein the cancer is prostate cancer, thyroid cancer, endocrine system cancer, brain cancer, breast cancer, cervical cancer, colon cancer, head and neck cancer, liver cancer, kidney cancer, lung cancer, non-small cell lung cancer, melanoma, mesothelioma, ovarian cancer, sarcoma, gastric cancer, uterine cancer, medulloblastoma, ampullary cancer, colorectal cancer, or pancreatic cancer.

[0346] 143. The method as described in item 131, wherein the cancer is prostate cancer.

[0347] 144. The method as described in item 131, wherein the cancer is ampullary carcinoma.

[0348] 145. The method as described in item 131, wherein the cancer is colorectal cancer.

[0349] 146. The method as described in item 131, wherein the cancer is lung cancer.

[0350] 147. The method as described in item 131, wherein the cancer is non-small cell lung cancer.

[0351] 148. The method as described in item 131, wherein the cancer is ovarian cancer.

[0352] 149. The method as described in item 131, wherein the cancer is pancreatic cancer.

[0353] 150. The method as described in Item 131, wherein the cancer is Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, neuroblastoma, glioma, glioblastoma multiforme, ovarian cancer, rhabdomyosarcoma, essential thrombocytosis, essential macroglobulinemia, primary brain tumor, cancer, malignant pancreatic insulinoma, malignant carcinoid tumor, bladder cancer, precancerous skin lesions, testicular cancer, lymphoma, thyroid cancer, neuroblastoma, esophageal cancer, urogenital tract cancer, malignant hypercalcemia, endometrial cancer, adrenocortical cancer, endocrine and exocrine pancreatic growths, medullary thyroid carcinoma, thyroid brain-like epithelial carcinoma, melanoma, colorectal cancer, papillary thyroid carcinoma, hepatocellular carcinoma, or prostate cancer.

[0354] 151. The method as described in item 129, wherein the subject is suffering from pre-malignant conditions and requires treatment.

[0355] abbreviation

[0356] This article uses abbreviations and terms commonly used in organic chemistry, medicinal chemistry, pharmacology, and medicine, and familiar to practitioners in these fields. Representative abbreviations and definitions are provided below:

[0357] Ac represents acetyl [CH3C(O)-], Ac2O represents acetic anhydride; AcOH represents acetic acid; APC represents antigen-presenting cells; aq. represents aqueous solution; 9-BBN represents 9-boronbicyclo[3.3.1]nonane; BINAP represents (2,2′-bis(diphenylphosphine)-1,1′-binaphthyl); Bn represents benzyl; BOC represents tert-butyloxycarbonyl; CDI represents carbonyl diimidazole; DCM represents dichloromethane; DIAD represents diisopropyl azodicarbonate; DIBAL represents diisobutylaluminum hydride; DIPEA represents diisopropylethylamine; DMA represents dimethylacetyl Amine; DMAP is 4-dimethylaminopyridine; DMF is N,N-dimethylformamide; DMSO is dimethyl sulfoxide; dppf is 1,1'-bis(diphenylphosphine)ferrocene; EDAC (or EDC) is 1-ethyl-3-[3-(dimethylamino)propyl]-carbodiimide hydrochloride; ESI is electrospray ionization mass spectrometry; Et2O is diethyl ether; Et3N is triethylamine; Et is ethyl; EtOAc is ethyl acetate; EtOH is ethanol; 3-F-Ph is 3-fluorophenyl; HATU is (1-[bis(dimethylamino)methylene]... ]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate; HCl is hydrochloric acid; HOBt is 1-hydroxybenzotriazole; HPLC is high performance liquid chromatography; LCMS is HPLC with mass spectrometry detection; LiHMDS is bis(trimethylsilyl)lithium; LG is leaving group; M is molar; mCPBA is m-chloroperbenzoic acid; mmol is millimolecular weight; Me is methyl; MeCN is acetonitrile; MeOH is methanol; Ms is methanesulfonyl; MS is mass spectrometry; N is normal; NaHMD S represents sodium hexamethyldisilamide; NaOAc represents sodium acetate; NaOtBu represents sodium tert-butoxide; NMO represents N-methylmorpholine N-oxide; NMP represents N-methylpyrrolidone; NMR represents nuclear magnetic resonance spectroscopy; Pd2(dba)3 represents tris(dibenzylacetone)dipalladium; PdCl2(PPh3)2 represents dichlorobis(triphenylphosphine)palladium; PG represents an unspecified protecting group; Ph represents phenyl; PhMe represents toluene; PPh3 represents triphenylphosphine; PMB represents p-methoxybenzyl; rt represents room temperature; RBF represents a round-bottom flask; RuPhos Pd G1 represents chloro-(2-dicyclohexylphosphine-2′,6′-diisopropoxy-1,1′-biphenyl)[2-(2-aminoethyl)phenyl]palladium(II); SEM represents [2-(trimethylsilyl)ethoxy]methyl; SFC represents supercritical fluid chromatography; S NAr represents nucleophilic aromatic substitution; TBAB represents tetrabutylammonium bromide; TBAF represents tetrabutylammonium fluoride; TBS represents tert-butyldimethylsilyl; tBu represents tert-butyl; Tf represents trifluoromethanesulfonic acid; TFA represents trifluoroacetic acid; THF represents tetrahydrofuran; THP represents tetrahydropyran; TLC represents thin-layer chromatography; TMAD represents tetramethylazodicarbonamide; TMS represents trimethylsilyl; TPAP represents tetrapropylammonium perruthenate; Ts represents p-toluenesulfonyl; UPLC represents ultra-high performance liquid chromatography.

[0358] definition

[0359] As used herein, the term "abnormal" means different from normal. When used to describe enzyme activity, abnormality refers to activity that is greater than or less than the average of normal or disease-free control samples. Abnormal activity can refer to a disease-causing level of activity, where restoring abnormal activity to a normal or disease-independent level (e.g., by administering a compound or method of use as described herein) results in the disease or a reduction in symptoms of one or more diseases. Abnormal activity can be measured by measuring changes in the substrate of the enzyme in question; a difference of 2-fold or greater in activity change can be considered abnormal. Abnormal activity can also refer to an increased dependence on a specific signaling pathway due to a deficiency in a single complementary pathway.

[0360] As used herein, the term "acyl" signifies a group –C(=O)–R, where R is an alkyl, alkenyl, ynyl, cycloalkyl, cycloalkenyl, cycloynyl, aryl, heteroaryl, or heterocyclic group. The acyl group may be optionally substituted, as described herein with respect to each corresponding R group.

[0361] As used herein, the term "adenocarcinoma" refers to a malignant tumor caused by glandular cells that line up along organs within an organism. Non-limiting examples of adenocarcinoma include non-small cell lung cancer, prostate cancer, pancreatic cancer, esophageal cancer, and colorectal cancer.

[0362] As used herein, the term "alkanoyl" refers to a hydrogen or alkyl group attached to a parent molecule via a carbonyl group, and is exemplified by formyl (i.e., formaldehyde group), acetyl, propionyl, butyryl, and isobutyryl. An unsubstituted alkanoyl group contains 1 to 7 carbons. As described herein with respect to alkyl groups, an alkanoyl group may be unsubstituted or substituted (e.g., optionally substituted C1-7 alkanoyl groups). A terminal "-acyl" may be added to another group defined herein, such as aryl, cycloalkyl, and heterocyclic groups, to define "aromatic acyl," "cycloalkanoyl," and "(heterocyclic)acyl." These groups represent carbonyl groups substituted with aryl, cycloalkyl, or heterocyclic groups, respectively. Each "aromatic acyl," "cycloalkanoyl," and "(heterocyclic)acyl" group may be optionally substituted, as defined herein with respect to "aryl," "cycloalkyl," or "heterocyclic," respectively.

[0363] As used herein, the term "alkenyl" refers to a non-cyclic monovalent straight-chain or branched hydrocarbon group containing one, two, or three carbon-carbon double bonds. Non-limiting examples of alkenyl groups include vinyl, propenyl, propenyl, 1-methylvinyl, butenyl, butenyl, butenyl, 3-methylpropenyl, 1-methylpropenyl, 2-methylpropenyl, and 1-methylpropenyl. Alkenyl groups may be optionally substituted as defined herein with respect to alkyl groups.

[0364] Unless otherwise specified, as used herein, the term "alkoxy" is expressed as – OR chemical substituent, where R is C 1-6 Alkyl group. In some embodiments, as defined herein, the alkyl group may be further substituted. The term “alkoxy” may be combined with other terms defined herein (e.g., aryl, cycloalkyl, or heterocyclic) to define “arylalkoxy,” “cycloalkylalkoxy,” and “(heterocyclic)alkoxy.” These groups respectively represent alkoxy groups substituted with aryl, cycloalkyl, or heterocyclic groups. As defined herein for each individual part, each “arylalkoxy,” “cycloalkylalkoxy,” and “(heterocyclic)alkoxy” may be optionally substituted.

[0365] As used herein, the term "alkoxyalkyl" represents a –L–O–R chemical substituent, where L is C. 1-6 Alkylene and R is C 1-6 Alkyl. Optionally substituted alkoxyalkyl is an alkoxyalkyl that has been optionally substituted as described herein with respect to alkyl.

[0366] Unless otherwise specified, as used herein, the term "alkyl" refers to a non-cyclic straight-chain or branched saturated hydrocarbon group having 1 to 12 carbons when unsubstituted. In some preferred embodiments, the unsubstituted alkyl group has 1 to 6 carbons. Alkyl groups are exemplified by the following: methyl; ethyl; n-propyl and isopropyl; n-butyl, sec-butyl, isobutyl, and tert-butyl; neopentyl, etc., and may optionally (where the valence allows) be substituted by one, two, three, or, in the case of an alkyl group having two or more carbons, four or more independently selected substituents from the group consisting of: amino, aryl, aryloxy, azide, cycloalkyl, cycloalkoxy, cycloalkenyl, cycloalkynyl, haloyl, heterocyclic, (heterocyclic)oxy, heteroaryl, hydroxyl, nitro, thiol, silyl, cyano, alkylsulfonyl, alkylsulfinyl, alkylsulfoxide, =O, =S, -SO2R (where R is amino or cycloalkyl), =NR' (where R' is H, alkyl, aryl, or heterocyclic). Each substituent may be unsubstituted itself or (where the valence allows) substituted with an unsubstituted substituent as defined herein for each corresponding group.

[0367] As used herein, the term "alkylene" refers to a divalent alkyl group. Optionally substituted alkylene groups are alkylene groups that have been optionally substituted as described herein with respect to alkyl groups.

[0368] As used herein, the term "alkylamino" refers to an amino group having the formula –N(R N1 )2 or –NHR N1 The group, wherein R is defined herein. N1 It is an alkyl group. As defined with respect to alkyl, the alkyl moiety of an alkylamino group may be optionally substituted. Each optional substituent on a substituted alkylamino group may be unsubstituted itself or (where the valence allows) substituted with an unsubstituted substituent as defined herein with respect to each corresponding group.

[0369] As used herein, the term "alkyl sulfide" represents a –S–(alkyl) group. As defined with respect to alkyl, the alkyl sulfide group may be optionally substituted.

[0370] As used herein, the term "alkylsulfinyl" represents the formula –S(O)–(alkyl) group. As defined with respect to alkyl, the alkylsulfinyl group may be optionally substituted.

[0371] As used herein, the term "alkylsulfonyl" represents the formula –S(O)2–(alkyl) group. As defined with respect to alkyl, the alkylsulfonyl group may be optionally substituted.

[0372] As used herein, the term "alkynyl" refers to a monovalent straight-chain or branched hydrocarbon group containing 2 to 6 carbon atoms and at least one carbon-carbon triple bond, and is illustrated by examples such as ethynyl, 1-propynyl, etc. As defined with respect to alkyl, alkynyl can be unsubstituted or substituted (e.g., optionally substituted alkynyl).

[0373] As used herein, the term "amino" represents –N(R N1 )2, where if the amino group is unsubstituted, then the two R groups are... N1 All are H; or, if the amino group is substituted, each R N1 Independently, it can be H, -OH, -NO2, or -N(R) N2 )2、-SO2OR N2 、-SO2R N2 -SOR N2 -COOR N2 N protecting group, alkyl, alkenyl, alkynyl, alkoxy, aryl, arylalkyl, aryloxy, cycloalkyl, cycloalkenyl, heteroalkyl or heterocyclic group, provided that at least one R N1 Not H, and each of R N2Independently, it can be H, alkyl, or aryl. Each substituent may be unsubstituted or substituted with an unsubstituted substituent as defined herein for each respective group. In some embodiments, the amino group is an unsubstituted amino group (i.e., -NH2) or a substituted amino group (e.g., NHR). N1 ), where R N1 Independently -OH, SO2OR N2 -SO2R N2 、-SOR N2 -COOR N2 Optionally substituted alkyl or Optionally substituted aryl, and each R N2 The amino group can be an optionally substituted alkyl group or an optionally substituted aryl group. In some embodiments, the substituted amino group can be an alkylamino group, wherein the alkyl group is optionally substituted as described herein with respect to alkyl. In some embodiments, the amino group is –NHR. N1 , where R N1 The alkyl group is optionally substituted.

[0374] As used herein, the term "aryl" refers to a monocyclic, bicyclic, or polycyclic carbocyclic system having one or two aromatic rings. An aryl group may contain 6 to 10 carbon atoms. All atoms within an unsubstituted carbocyclic aryl group are carbon atoms. Non-limiting examples of carbocyclic aryl groups include phenyl, naphthyl, 1,2-dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, fluorenyl, indene, indene, etc. An aryl group may be unsubstituted or substituted with 1, 2, 3, 4, or 5 substituents independently selected from the group consisting of: alkyl, alkenyl, alkynyl, alkoxy, alkylsulfinyl, alkylsulfonyl, alkylsulfonyl, amino, aryl, aryloxy, azide, cycloalkyl, cycloalkoxy, cycloalkenyl, cycloalkynyl, cycloalkenyl, cycloalkynyl, haloyl, heteroalkyl, heterocyclic, (heterocyclic)oxy, hydroxyl, nitro, thiol, silyl, and cyano. Each substituent may be unsubstituted or substituted by an unsubstituted substituent as defined herein for each respective group.

[0375] As used herein, the term "arylalkyl" refers to an alkyl group substituted with an aryl group. As described herein, the aryl and alkyl moieties may be optionally substituted as separate groups.

[0376] As used herein, the term "arylene" refers to a divalent aryl group. Optionally substituted arylene groups are those that have been optionally substituted as described herein with respect to aryl groups.

[0377] Unless otherwise specified, as used herein, the term "alkoxy" is expressed as – OR chemical substituent, where R is aryl. In optionally substituted aryloxy groups, as described herein with respect to aryl, the aryl group is optionally substituted.

[0378] As used herein, the term "ATR inhibitor" refers to the reduction of ATR kinase activity upon contact with the enzyme ATR kinase, whether in vitro, in cell culture, or in vivo in animals, to the extent that the measured ATR kinase IC50 value is reduced. 50 Compounds with a concentration of 10 μM or less (e.g., 5 μM or less, or 1 μM or less). For some ATR inhibitors, ATR kinase IC50... 50 It can be 100 nM or less (e.g., 10 nM or less or 1 nM or less) and can be as low as 100 pM or 10 pM. Preferably, ATR kinase IC 50 It ranges from 1 nM to 1 μM (e.g., 1 nM to 750 nM, 1 nM to 500 nM, or 1 nM to 250 nM).

[0379] As used in this article, the term "ATR kinase" refers to ataxia-telangiectasia and RAD-3-related protein kinase.

[0380] As used in this article, the term "azido" refers to the -N3 group.

[0381] As used herein, the term "cancer" refers to all types of cancer, growths, or malignant tumors found in mammals (such as humans), including leukemia, epithelial carcinoma, and sarcoma. Non-limiting examples of cancers that can be treated with the compounds or methods provided herein include prostate cancer, thyroid cancer, endocrine system cancers, brain cancer, breast cancer, cervical cancer, colon cancer, head and neck cancer, liver cancer, kidney cancer, lung cancer, non-small cell lung cancer, melanoma, mesothelioma, ovarian cancer, sarcoma, gastric cancer, uterine cancer, medulloblastoma, ampullary cancer, colorectal cancer, and pancreatic cancer. Other non-limiting examples may include Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, neuroblastoma, glioma, glioblastoma multiforme, ovarian cancer, rhabdomyosarcoma, essential thrombocytosis, essential macroglobulinemia, primary brain tumors, cancer, malignant pancreatic insulinoma, malignant carcinoid tumor, bladder cancer, precancerous skin lesions, testicular cancer, lymphoma, thyroid cancer, neuroblastoma, esophageal cancer, urogenital tract cancer, malignant hypercalcemia, endometrial cancer, adrenocortical carcinoma, endocrine and exocrine pancreatic vegetations, medullary thyroid carcinoma, thyroid brain-like epithelial carcinoma, melanoma, colorectal cancer, papillary thyroid carcinoma, hepatocellular carcinoma, and prostate cancer.

[0382] As used herein, the term "carbocyclic" refers to an optionally substituted C3-16 monocyclic, bicyclic, or tricyclic structure in which the ring, which may be aromatic or non-aromatic, is formed from carbon atoms. Carbocyclic structures include cycloalkyl, cycloalkenyl, cycloynyl, and certain aryl groups.

[0383] As used in this article, the term "carbonyl" refers to a –C(O)– group.

[0384] As used herein, the term "epithelial carcinoma" refers to a malignant new growth composed of epithelial cells that tend to infiltrate surrounding tissues and metastasize. Non-limiting examples of epithelial carcinomas that can be treated with the compounds or methods provided herein include, for example, thyroid brain-like epithelial carcinoma, familial thyroid brain-like epithelial carcinoma, acinar epithelial carcinoma, acinar epithelial carcinoma, adenocystic epithelial carcinoma, adenoid cystic epithelial carcinoma, adenomatous epithelial carcinoma, adrenocortical epithelial carcinoma, alveolar epithelial carcinoma, alveolar cell epithelial carcinoma, basal cell epithelial carcinoma, basal cell carcinoma, basaloid epithelial carcinoma, basal squamous cell carcinoma, bronchioloalveolar epithelial carcinoma, bronchiolar epithelial carcinoma, bronchial epithelial carcinoma, medullary epithelial carcinoma, cholangiocarcinoma, and choriocarcinoma. Colloidal carcinoma, comedo-like carcinoma, main carcinoma, cribriform carcinoma, armored carcinoma, cutaneous carcinoma, columnar carcinoma, columnar cell carcinoma, ductal carcinoma, sclerothelial carcinoma, embryonal carcinoma, cerebral carcinoma, epidermoid carcinoma, adenoid carcinoma, exophytic carcinoma, ulcerative carcinoma, fibrocarcinoma, colloidal carcinoma, gelatinous carcinoma, giant cell carcinoma, giant cell carcinoma, adenocarcinoma, granular cell carcinoma, pilostromal carcinoma, polyangioid carcinoma, hepatocellular carcinoma, Schulte's cell carcinoma, hyaline carcinoma, adrenoid carcinoma Cancer, infantile embryonic epithelial carcinoma, epithelial carcinoma in situ, epithelial intraepithelial carcinoma, intraepithelial carcinoma, Klinefelter's cell carcinoma, Kurtschitzky cell carcinoma, large cell carcinoma, lenticular carcinoma, lenticular epithelial carcinoma, lipomatous carcinoma, lymphoepithelial carcinoma, medullary carcinoma, medullary carcinoma, melanoma, soft epithelial carcinoma, mucinous carcinoma, mucinous cell carcinoma, mucinous epidermoid carcinoma, mucinous carcinoma, mucinous epithelial carcinoma, myxomatous carcinoma, nasopharyngeal carcinoma, oat cell carcinoma, ossifying epithelial carcinoma, osteoid carcinoma, papillary carcinoma, periportal epithelium Carcinoma, invasive preepithelial carcinoma, acanthosis nigra carcinoma, soft pasty carcinoma, renal cell carcinoma, reservoir cell carcinoma, sarcomatoid carcinoma, Schneider's carcinoma, sclerosing carcinoma, scrotal carcinoma, signet ring cell carcinoma, simple carcinoma, small cell carcinoma, potato-shaped carcinoma, spherical cell carcinoma, spindle cell carcinoma, cavernous carcinoma, squamous carcinoma, squamous cell carcinoma, string carcinoma, capillary carcinoma, angiotensinus carcinoma, transitional cell carcinoma, nodular carcinoma, nodular epithelial carcinoma, verrucous carcinoma, and villous carcinoma.

[0385] As used in this article, the term "cyano" refers to the –CN group.

[0386] Unless otherwise specified, as used herein, the term "cycloalkenyl" refers to a non-aromatic carbocyclic group having at least one intracyclic double bond and 3 to 10 carbons (e.g., C10, C20, C30, C40, C50, C60, C7 ...60, C 3-10(Cycloalkenyl). Non-limiting examples of cycloalkenyl include cyclopropenyl-1-enyl, cyclopropenyl-2-enyl, cyclobutenyl-1-enyl, cyclobutenyl-1-enyl, cyclobutenyl-2-enyl, cyclopentenyl-1-enyl, cyclopentenyl-2-enyl, cyclopentenyl-3-enyl, norbornenyl-1-enyl, norbornenyl-2-enyl, norbornenyl-5-enyl, and norbornenyl-7-enyl. As described with respect to cycloalkyl, the cycloalkenyl may be unsubstituted or substituted (e.g., optionally substituted cycloalkenyl).

[0387] As used herein, the term "cycloalkenylalkyl" refers to an alkyl group substituted with a cycloalkenyl group, each as defined herein. The cycloalkenyl and alkyl moieties may be substituted with individual groups as defined herein.

[0388] Unless otherwise specified, as used herein, the term "alkoxy" signifies – OR chemical substituent, where R is a cycloalkyl group. In some embodiments, as defined herein, the cycloalkyl group may be further substituted.

[0389] Unless otherwise specified, as used herein, the term "cycloalkyl" refers to a cycloalkyl group having 3 to 10 carbon atoms (e.g., C10, C20, C30, C40, C50, C60, C7 ... 3-10 Cycloalkyl groups can be monocyclic or bicyclic. Bicyclic cycloalkyl groups can be of the bicyclic [pq0]alkyl type, wherein each p and q is independently 1, 2, 3, 4, 5, 6, or 7, provided that the sum of p and q is 2, 3, 4, 5, 6, 7, or 8. Alternatively, bicyclic cycloalkyl groups can comprise bridged cycloalkyl structures, for example, bicyclic [pqr]alkyl, wherein r is 1, 2, or 3, and each p and q is independently 1, 2, 3, 4, 5, or 6, provided that the sum of p, q, and r is 3, 4, 5, 6, 7, or 8. Cycloalkyl groups can be spirocyclic, for example, spiro[pq]alkyl, wherein each p and q is independently 2, 3, 4, 5, 6, or 7, provided that the sum of p and q is 4, 5, 6, 7, 8, or 9. Non-limiting examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, 1-bicyclo[2.2.1.]heptyl, 2-bicyclo[2.2.1.]heptyl, 5-bicyclo[2.2.1.]heptyl, 7-bicyclo[2.2.1.]heptyl and decalinyl. The cycloalkyl group may be unsubstituted or substituted by 1, 2, 3, 4 or 5 substituents independently selected from the group consisting of: alkyl, alkenyl, alkynyl, alkoxy, alkylsulfinyl, alkylsulfonyl, alkylsulfonyl, amino, aryl, aryloxy, azide, cycloalkyl, cycloalkoxy, cycloalkenyl, cycloalkynyl, haloyl, heteroalkyl, heterocyclic, (heterocyclic)oxy, heteroaryl, hydroxyl, nitro, thiol, silyl, cyano, =O, =S, -SO2R (where R is amino or cycloalkyl), =NR' (where R' is H, alkyl, aryl or heterocyclic) or –CON(R A)2 (where each R A Independently H or alkyl or two R A Together with the atoms to which they are attached, they combine to form a heterocyclic group. Each substituent may be unsubstituted or substituted by an unsubstituted substituent as defined herein for each corresponding group.

[0390] As used herein, the term "cycloalkylalkyl" refers to an alkyl group substituted with a cycloalkyl group, each as defined herein. The cycloalkyl and alkyl moiety may be optionally substituted as separate groups as described herein.

[0391] As used herein, the term "cycloalkylene" refers to a divalent cycloalkyl group. Optionally substituted cycloalkylene groups are those that have been optionally substituted as described herein with respect to cycloalkyl groups.

[0392] Unless otherwise stated, as used herein, the term "cycloynyl" refers to a monovalent carbocyclic group having one or two carbon-carbon triple bonds and having 8 to 12 carbon atoms. A cycloynyl group may contain a transcyclic bond or bridge. Non-limiting examples of cycloynyl groups include cyclooctyynyl, cyclononyynyl, cyclodecynyl, and cyclodecadiynyl. As defined with respect to cycloalkyl groups, a cycloynyl group may be unsubstituted or substituted (e.g., optionally substituted cycloynyl groups).

[0393] "Disease" or "symptom" refers to the state or health condition of a patient or subject who can be treated with the compounds or methods provided herein.

[0394] As used herein, the term "halogen" refers to a halogen selected from bromine, chlorine, iodine, and fluorine.

[0395] As used herein, the term "heteroalkyl" refers to an alkyl, alkenyl, or ynyl group consisting of one or two heteroatoms spaced once, twice, three times, or four times independently of each other. Each heteroatom is independently O, N, or S. In some embodiments, the heteroatom is O or N. No heteroalkyl group contains two consecutive oxygen or sulfur atoms. Heteroalkyl groups can be unsubstituted or substituted (e.g., optionally substituted heteroalkyl groups). When the heteroalkyl group is substituted and the substituent is bonded to the heteroatom, the substituent is selected based on the nature and valence of the heteroatom. Therefore, the substituents bonded to the heteroatom (where the valence allows) are selected from the group consisting of: =O, -N(R N2 )2、-SO2OR N3 -SO2R N2 、-SOR N3 -COOR N3 ,N protecting group, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, cycloalkenyl, cycloalkynyl, heterocyclic or cyano, wherein each R N2Independently H, alkyl, cycloalkyl, cycloalkenyl, cycloynyl, aryl, or heterocyclic, and each R N3 Independently, they are alkyl, cycloalkyl, cycloalkenyl, cycloynyl, aryl, or heterocyclic groups. Each of these substituents may be unsubstituted itself or substituted by an unsubstituted substituent as defined herein for each respective group. When the heteroalkyl group is substituted and the substituent is bonded to a carbon atom, the substituent is selected from those described for the alkyl group, provided that the substituent bonded to the carbon atom to the heteroatom is not Cl, Br, or I. It should be understood that the carbon atom is at the end of the heteroalkyl group.

[0396] As used herein, the term "heteroarylalkyl" refers to an alkyl group substituted with a heteroaryl group, each as defined herein. The heteroaryl and alkyl moiety may be optionally substituted as separate groups as described herein.

[0397] As used herein, the term "hybrid aryl" refers to a divalent heteroaryl. Optionally substituted heteroaryl is a heteroaryl that has been optionally substituted as described herein with respect to heteroaryl.

[0398] As used herein, the term "heteroaryloxy group" refers to the structure – OR, in which R is a heteroaryl group. As defined with respect to heterocyclic groups, the heteroaryloxy group may be optionally substituted.

[0399] As used herein, the term "heterocyclic group" refers to a monocyclic, bicyclic, tricyclic, or tetracyclic system having a fused, bridged, or spirocyclic 3, 4, 5, 6, 7, or 8-membered ring, wherein, unless otherwise specified, the ring comprises 1, 2, 3, or 4 heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. In some embodiments, "heterocyclic group" is a monocyclic, bicyclic, tricyclic, or tetracyclic system having a fused or bridged 5, 6, 7, or 8-membered ring, wherein, unless otherwise specified, the ring comprises 1, 2, 3, or 4 heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. Heterocyclic groups can be aromatic or non-aromatic. Non-aromatic 5-membered heterocyclic groups have 0 or 1 double bonds, non-aromatic 6- and 7-membered heterocyclic groups have 0 to 2 double bonds, and non-aromatic 8-membered heterocyclic groups have 0 to 2 double bonds and / or 0 or 1 carbon-carbon triple bonds. Unless otherwise specified, heterocyclic groups comprise 1 to 16 carbon atoms. Some heterocyclic groups can contain up to nine carbon atoms. Non-aromatic heterocyclic groups include pyrrolinyl, pyrrolylalkyl, pyrazolinyl, pyrazolylalkyl, imidazolinyl, imidazolinyl, piperidinyl, homopiperidinyl, piperazinyl, pyridazinyl, oxazolinyl, isoxazolinyl, morpholinyl, thiomorpholinyl, thiazolinyl, isothiazolinyl, thiazolinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, dihydroindolyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, pyranyl, dihydropyranyl, dithiazolinyl, etc. If the heterocyclic system has at least one aromatic resonance structure or at least one aromatic tautomer, then such a structure is an aromatic heterocyclic group (i.e., a heteroaryl). Non-limiting examples of heteroaryl groups include benzimidazolyl, benzofuranyl, benzothiazolyl, benzothiophene, benzooxazolyl, furanyl, imidazolyl, indolyl, isoyindolyl, isoquinolinyl, isothiazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, purine, pyrroleyl, pyridyl, pyrazinyl, pyrimidinyl, quinazolinyl, quinolinyl, thiadiazolyl (e.g., 1,3,4-thiadiazolyl), thiazolyl, thiophene, triazolyl, tetrazolyl, etc. The term "heterocyclic group" also refers to a heterocyclic compound having a bridging polycyclic structure in which one or more carbon atoms and / or heteroatoms bridge two non-adjacent members of a monocyclic ring (e.g., quinine, tropane, or diazabicyclo[2.2.2]octane). The term "heterocyclic group" includes bicyclic, tricyclic, and tetracyclic groups in which any of the heterocycles mentioned above is fused with one, two, or three carbocyclic rings, such as an aryl ring, a cyclohexane ring, a cyclohexene ring, a cyclopentane ring, a cyclopentene ring, or another monocyclic heterocycle. Examples of fused heterocycles include 1,2,3,5,8,8a-hexahydroindoleazine, 2,3-dihydrobenzofuran, 2,3-dihydroindole, and 2,3-dihydrobenzothiophene.The heterocyclic group may be unsubstituted or substituted by 1, 2, 3, 4 or 5 substituents independently selected from the group consisting of: alkyl, alkenyl, alkynyl, alkoxy, alkylsulfinyl, alkylsulfonyl, alkylsulfonyl, amino, aryl, aryloxy, azide, cycloalkyl, cycloalkoxy, cycloalkenyl, cycloalkynyl, haloyl, heteroalkyl, heterocyclic, (heterocyclic)oxy, hydroxyl, nitro, thiol, silyl, cyano, =O, =S, =NR' (where R' is H, alkyl, aryl or heterocyclic). Each substituent itself may be unsubstituted or substituted by an unsubstituted substituent as defined herein for each corresponding group.

[0400] As used herein, the term "heterocyclic alkyl" refers to an alkyl group substituted with a heterocyclic group, each as defined herein. The heterocyclic and alkyl moieties may be optionally substituted as separate groups as described herein.

[0401] As used herein, the term "subheterocyclic group" refers to a divalent heterocyclic group. Optionally substituted subheterocyclic groups are optionally substituted subheterocyclic groups as described herein with respect to heterocyclic groups.

[0402] Unless otherwise specified, as used herein, the term "(heterocyclic)oxy group" is expressed as – OR chemical substituent, where R is a heterocyclic group. The (heterocyclic)oxy group may be optionally substituted in the manner described for the heterocyclic group.

[0403] As used interchangeably in this document, the term "hydroxyl (hydroxyl and hydroxy)" refers to the -OH group.

[0404] As used herein, the term "isotope-enriched" refers to a pharmaceutically active agent in which the isotope at a predetermined position within the molecule is at least 100 times more abundant than the natural abundance of that isotope. For example, a composition enriched with deuterium contains an active agent in which the abundance of deuterium at at least one hydrogen atom position is at least 100 times greater than the natural abundance of deuterium. Preferably, the deuterium enrichment is at least 1000 times greater than the natural abundance of deuterium. More preferably, the deuterium enrichment is at least 4000 times greater than the natural abundance of deuterium (e.g., at least 4750 times, e.g., up to 5000 times).

[0405] As used herein, the term “leukemia” broadly refers to a progressive malignant disease of the hematopoietic organs and is typically characterized by the abnormal proliferation and development of white blood cells and their precursors in the blood and bone marrow. Leukemia is usually clinically classified based on the following: (1) duration and characteristics of the disease: acute or chronic; (2) cell types involved: myeloid, lymphoid, or monocytic; and (3) whether the number of abnormal cells in the blood is increased or not increased in leukemia or non-leukemia (subleukemia). Exemplary leukemias that can be treated with the compounds or methods provided herein include, for example, acute non-lymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, acute promyelocytic leukemia, adult T-cell leukemia, non-leukemic leukemia, leukemic leukemia, basophilic leukemia, blastic leukemia, bovine leukemia, chronic myeloid leukemia, cutaneous leukemia, embryonic leukemia, eosinophilic leukemia, Gross's leukemia, hairy cell leukemia, hematopoietic leukemia, hematopoietic leukemia, histiocytic leukemia, stem cell leukemia, and acute monocytic leukemia. Leukopenic leukemia, lymphocytic leukemia, lymphoblastic leukemia, lymphoblastic leukemia, lymphoid leukemia, lymphosarcoma cell leukemia, mast cell leukemia, megakaryocytic leukemia, micromyeloid leukemia, monocytic leukemia, myeloblastic leukemia, myeloid leukemia, myeloid granulocytic leukemia, granulocytic-monocytic leukemia, Neghly leukemia, plasma cell leukemia, multiple myeloma, plasma cell leukemia, promyelocytic leukemia, Riddle cell leukemia, Schilling leukemia, stem cell leukemia, subleukemic leukemia, and undifferentiated cell leukemia.

[0406] As used herein, the term “lymphoma” refers to cancer caused by immune-originating cells. Non-limiting examples of T-cell and B-cell lymphomas include non-Hodgkin's lymphoma and Hodgkin's disease, diffuse large B-cell lymphoma, follicular lymphoma, mucosa-associated lymphoid tissue (MALT) lymphoma, small cell lymphocytic lymphoma-chronic lymphocytic leukemia, mantle cell lymphoma, mediastinal (thymic) large B-cell lymphoma, lymphoplasmacytic lymphoma-Wald's macroglobulinemia, peripheral T-cell lymphoma (PTCL), angioimmunoblastic T-cell lymphoma (AITL) / follicular T-cell lymphoma (FTCL), anaplastic large cell lymphoma (ALCL), enteropathy-associated T-cell lymphoma (EATL), adult T-cell leukemia / lymphoma (ATLL), or extranodal NK / T-cell lymphoma (nasal type).

[0407] As used herein, the term "melanoma" refers to a tumor arising from the melanocyte system of the skin and other organs. Melanomas that can be treated with the compounds or methods provided herein include, for example, acral-lentiginous melanoma, amelanoma, benign juvenile melanoma, Claudemann melanoma, S91 melanoma, Hartmann-Parkinson's disease melanoma, juvenile melanoma, malignant lentiginous melanoma, malignant melanoma, nodular melanoma, subungual melanoma, and superficial diffuse melanoma.

[0408] As used in this article, the term "nitro" refers to a –NO2 group.

[0409] As used herein, the term "oxo" refers to a divalent oxygen atom (e.g., an oxo structure can be represented as =O).

[0410] As used in this article, the term "phenyl" refers to phenyl.

[0411] As used herein, the term "pharmaceutical composition" means a composition containing the compounds described herein formulated together with pharmaceutically acceptable excipients and manufactured or marketed with the approval of a government regulatory agency as part of a treatment regimen for treating a disease in mammals. Pharmaceutical compositions may be formulated, for example, for oral administration in unit dosage forms (e.g., tablets, capsules, pouches, soft capsules, or syrups); for topical administration (e.g., in the form of creams, gels, lotions, or ointments); for intravenous administration (e.g., in the form of a sterile solution without microparticle plugs and in a solvent system suitable for intravenous use); or as any other formulation described herein.

[0412] As used interchangeably herein, the terms "pharmaceuticalally acceptable excipient" or "pharmaceuticalally acceptable carrier" refer to any component other than the compounds described herein that has properties of being non-toxic and non-inflammatory in a patient (e.g., a medium capable of suspending or dissolving an active compound). Excipients may include, for example: anti-adhesives, antioxidants, adhesives, coatings, compression aids, disintegrants, dyes (colorants), softeners, emulsifiers, fillers (diluents), film-forming agents or coatings, flavorings, fragrances, flow enhancers (flow improvers), lubricants, preservatives, printing inks, adsorbents, suspending or dispersing agents, sweeteners, or hydrates. Exemplary excipients include, but are not limited to: butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, croscarmellose, citric acid, crospovidone, cysteine, ethyl cellulose, gelatin, hydroxypropyl cellulose, hydroxypropyl methylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methylparaben, microcrystalline cellulose, polyethylene glycol, polyvinylpyrrolidone, povidone, pregelatinized starch, propylparaben, retinyl palmitate, shellac, silica, sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol.

[0413] As used herein, the term "pharmaceutically acceptable salt" refers to those salts that, within reasonable medical judgment, are suitable for contact with tissues in humans and animals without excessive toxicity, irritation, anaphylactic reactions, 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 Berge et al., J. Pharmaceutical Sciences 66:1-19, 1977 and Pharmaceutical Salts: Properties, Selection, and Use, (edited by P.H. Stahl and C. G. Germuth), Wiley-VCH, 2008. Salts may be prepared in situ during the final isolation and purification of the compounds described herein or by reacting the free base group with a suitable organic acid. Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, hydrogen sulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, gluconate, dodecyl sulfate, ethanesulfonate, fumarate, glucono-p-ethylhexanoate, glyceryl phosphate, hemisulfate, heptanate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, dihydroxynaphthalate, pectate, persulfate, 3-phenylpropionate, phosphate, picrate, neopentanoate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate, etc. Representative alkali metal salts or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc., as well as non-toxic ammonium, quaternary ammonium and amine cations, including but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, etc.

[0414] As used herein, the terms “pre-malignant” or “precancerous” refer to conditions that are not malignant but could become malignant at any time. Non-limiting examples of pre-malignant conditions include myelodysplastic syndromes, colonic polyps, actinic keratosis, cervical dysplasia, pulmonary metaplasia, and leukoplakia.

[0415] As used herein, the term "protecting group" refers to a group intended to protect a hydroxyl, amino, or carbonyl group from participating in one or more undesirable reactions during chemical synthesis. As used herein, the term "O-protecting group" refers to a group intended to protect a hydroxyl or carbonyl group from participating in one or more undesirable reactions during chemical synthesis. As used herein, the term "N-protecting group" refers to a group intended to protect a nitrogen-containing (e.g., amino, amide, heterocyclic NH, or hydrazine) group from participating in one or more undesirable reactions during chemical synthesis. Commonly used O-protecting and N-protecting groups are disclosed in Greene, "Protective Groups in Organic Synthesis," 3rd edition (John Wiley & Sons, New York, 1999), which is incorporated herein by reference. Exemplary O-protecting groups and N-protecting groups include alkyl, aromatic, or carbamoyl groups, such as formyl, acetyl, propionyl, neopentanoyl, tert-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthaloyl, o-nitrophenoxyacetyl, α-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, tert-butyldimethylsilyl, tri-isopropylsilyloxymethyl, 4,4'-dimethoxytriphenylmethyl, isobutyryl, phenoxyacetyl, 4-isopropylphenoxyacetyl, dimethylformamidinyl, and 4-nitrobenzoyl.

[0416] Exemplary O-protecting groups for protecting carbonyl groups include, but are not limited to: acetal, acyl, 1,3-dithiaane, 1,3-dioxane, 1,3-dioxolane, and 1,3-dithiopentane.

[0417] Other O-protecting groups include, but are not limited to: substituted alkyl, aryl, and arylalkyl ethers (e.g., triphenylmethyl, methylthiomethyl, methoxymethyl, benzyloxymethyl, silyloxymethyl, 2,2,2-trichloroethoxymethyl, tetrahydropyranyl, tetrahydrofuranyl, ethoxyethyl, 1-[2-(trimethylsilyl)ethoxy]ethyl, 2-trimethylsilylethyl, tert-butyl ether, p-chlorophenyl, p-methoxyphenyl, p-nitrophenyl, benzyl, p-methoxybenzyl, and nitrobenzyl); silyl ethers (e.g., Trimethylsilyl, triethylsilyl, triisopropylsilyl, dimethylisopropylsilyl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, tribenzylsilyl, triphenylsilyl and diphenylmethylsilyl); carbonates (e.g. methyl, methoxymethyl, 9-fluorenylmethyl, ethyl, 2,2,2-trichloroethyl, 2-(trimethylsilyl)ethyl, vinyl, allyl, nitrophenyl, benzyl, methoxybenzyl, 3,4-dimethoxybenzyl and nitrobenzyl).

[0418] Other N-protecting groups include, but are not limited to: chiral auxiliaries, such as protected or unprotected D-amino acids, L-amino acids, or D,L-amino acids (such as alanine, leucine, phenylalanine, etc.); sulfonyl groups, such as benzylsulfonyl, p-toluenesulfonyl, etc.; carbamate forming groups, such as benzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl, 2,4-dimethoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl, 3,4,5-trimethoxybenzyloxycarbonyl, 1-(p-biphenyl) (2,4-Dimethyl-3,5-dimethoxybenzyloxycarbonyl, benzoyloxycarbonyl, tert-butoxycarbonyl, diisopropylmethoxycarbonyl, isopropoxycarbonyl, ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl, cyclopentoxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl, phenylthiocarbonyl, etc.); arylalkyl groups, such as benzyl, p-methoxybenzyl, 2,4-dimethoxybenzyl, triphenylmethyl, benzyloxymethyl, etc.; silyl acetal groups, such as [2-(trimethylsilyl)ethoxy]methyl); and silyl groups, such as trimethylsilyl, etc. Available N-protecting groups include formyl, acetyl, benzoyl, neopentyl, tert-butylacetyl, alanyl, benzyl, dimethoxybenzyl, [2-(trimethylsilyl)ethoxy]methyl (SEM), tetrahydropyranyl (THP), tert-butyloxycarbonyl (Boc), and benzyloxycarbonyl (Cbz).

[0419] The term "tautomer" refers to structural isomers that are readily interchangeable through proton migration. Tautomers are different chemical species that can be identified by different spectroscopic features, but are usually not separable individually. Non-limiting examples of tautomers include keto-enols, enamine-imines, amide-imines, nitroso-oximes, enone-alkynols, and amino acid-ammonium formates.

[0420] The term "sarcoma" generally refers to a tumor composed of embryonic connective tissue-like material and typically consisting of densely packed cells embedded in fibrous or homogeneous material. Non-limiting examples of sarcomas treatable with the compounds or methods described herein include, for example, chondrosarcoma, fibrosarcoma, lymphosarcoma, melanosarcoma, myxosarcoma, osteosarcoma, Ebernay's sarcoma, liposarcoma, liposarcoma, alveolar soft tissue sarcoma, ameloblastic sarcoma, botryoid sarcoma, chloroma sarcoma, choriocarcinoma, embryonal sarcoma, Williams' tumor sarcoma, endometrial sarcoma, stromal sarcoma, Ewing's sarcoma, and fascial sarcoma. Fibroblastic sarcoma, giant cell sarcoma, granulocyte sarcoma, Hodgkin's sarcoma, idiopathic multiple pigmented hemorrhagic sarcoma, B-cell immunoblastic sarcoma, T-cell immunoblastic sarcoma, Jensen's sarcoma, Kaposi's sarcoma, Kupffer cell sarcoma, angiosarcoma, leukemic sarcoma, malignant mesenchymal sarcoma, paraosteal sarcoma, reticulum cell sarcoma, Rouss's sarcoma, serous cystic sarcoma, synovial sarcoma, and capillary hemangiosarcoma.

[0421] As used herein, the term "subject" means a human or non-human animal (e.g., a mammal) that is determined by a qualified professional (e.g., a physician or nursing practitioner) to have or be at risk of a disease or symptom, with or without laboratory testing known in the art on samples from the subject. Preferably, the subject is a human. Non-limiting examples of diseases and symptom include diseases characterized by excessive cell proliferation, such as cancer.

[0422] As used herein, the term "treatment" refers to the medical management of a subject with an intention to improve, enhance, stabilize, prevent, or cure a disease or symptom. This term includes active treatment (treatment involving the improvement of a disease or symptom), etiological treatment (treatment involving the cause of a related disease or symptom), palliative treatment (treatment designed to alleviate the symptoms of a disease or symptom), preventive treatment (treatment involving minimizing or partially or completely suppressing the development of a related disease or symptom), and supportive treatment (treatment used to complement another therapy). Detailed Implementation

[0423] Generally, this invention provides compounds, pharmaceutical compositions comprising the compounds, methods for preparing the compounds, and methods of use. The compounds of this invention may be ATR kinase inhibitors. These compounds can be used to inhibit ATR kinases in cells (e.g., cells of a subject). The subject may need treatment for a disease or condition, such as a disease or condition characterized by excessive cell proliferation, such as cancer. The ATR kinase inhibitory activity of the compounds disclosed herein can be used to treat subjects who require cancer treatment. Non-limiting examples of cancers that can be treated with the compounds disclosed herein are provided in Foote et al., J. Med. Chem., 61:9889-9907, 2018; Wengner et al., Mol. Cancer Ther., doi:10.1158 / 1535-7163.MCT-19-0019; and Dillon and Harrington, “Targeting ATR for Cancer Therapy: ATR-Targeted Drug Candidates”, Targeting the DNA Damage Response for Anti-Cancer Therapy, Pollard and Curtin, eds.; Humana Press, Cham (2018), pp. 99-127.

[0424] This invention provides a compound of formula (I):

[0425]

[0426] Or its pharmaceutically acceptable salt.

[0427] in

[0428] It is a double bond, and each Y is independently either N or CR. 4 ;or It is a single bond, and each Y is independently NR. Y carbonyl or C(R) Y )2; where each R Y Independently H or optionally substituted C 1-6 alkyl;

[0429] R 1 C is an optional replacement 1-6 Alkyl or H;

[0430] R 2 C is an optional replacement 2-9 Heterocyclic groups, optionally substituted C 1-6 Alkyl, optionally substituted C 3-8 cycloalkyl, optionally substituted C 2-9 Heterocyclic C1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 heteroaryl, optionally substituted C 1-9 heteroaryl C 1-6 Alkyl, halogen, –N(R) 5 2. –OR 5 –CON(R) 6 )2、–SO2N(R 6 )2、–SO2R 5A Or –Q–R 5B ;

[0431] R 3 C is an optional replacement 1-9 heteroaryl or optionally substituted C 1-9 heteroaryl C 1-6 alkyl;

[0432] Each R 4 Independently hydrogen, halogen, or optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 alkenyl or optionally substituted C 2-6 alkynyl group;

[0433] Each R 5 Independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 heteroaryl or –SO2R 5A ; or two Rs 5 Together with the atoms it is attached to, they combine to form optionally substituted C atoms. 2-9 Heterocyclic groups;

[0434] Each R 5A C can be substituted independently. 1-6 Alkyl, optionally substituted C 3-8 cycloalkyl or optionally substituted C 6-10 aryl;

[0435] R 5B Hydroxyl group, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 heteroaryl, –N(R) 5 )2、–CON(R 6 )2、–SO2N(R 6 )2、–SO2R 5A Or optionally substituted alkoxy groups;

[0436] Each R 6 Independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkoxyalkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 3-8 cycloalkyl or optionally substituted C 1-9 heteroaryl; or two R 6 Together with the atoms it is attached to, they combine to form optionally substituted C atoms. 2-9 Heterocyclic groups;

[0437] Q is an optional substitution for C. 2-9 Heterocyclic groups, optionally substituted C 3-8 Cycloalkylene, optionally substituted C 1-9 heteroaryl or optionally substituted C 6-10 Alpha-aryl; and

[0438] X is hydrogen or halogen.

[0439] The compounds of this invention may be, for example, compounds of formula (II):

[0440]

[0441] or a pharmaceutically acceptable salt thereof,

[0442] in

[0443] Each Y is independently either N or CR 4 ;

[0444] R 1 C is an optional replacement 1-6 Alkyl or H;

[0445] R 2 C is an optional replacement 2-9 Heterocyclic groups, optionally substituted C 1-6 Alkyl, optionally substituted C 3-8 cycloalkyl, optionally substituted C 2-9 Heterocyclic C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 Heteroaryl, optionally substituted C 1-9 heteroaryl C 1-6 Alkyl, halogen, –N(R) 5 2. –OR 5 –CON(R) 6 )2、–SO2N(R 6 )2、–SO2R 5AOr –Q–R 5B ;

[0446] R 3 C is an optional replacement 1-9 heteroaryl or optionally substituted C 1-9 heteroaryl C 1-6 alkyl;

[0447] Each R 4 Independently hydrogen, halogen, or optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 alkenyl or optionally substituted C 2-6 alkynyl group;

[0448] Each R 5 Independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 heteroaryl or –SO2R 5A ; or two Rs 5 Together with the atoms it is attached to, they combine to form optionally substituted C atoms. 2-9 Heterocyclic groups;

[0449] Each R 5A C can be substituted independently. 1-6 Alkyl, optionally substituted C 3-8 cycloalkyl or optionally substituted C 6-10 aryl;

[0450] R 5B Hydroxyl group, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 heteroaryl, –N(R) 5 )2、–CON(R 6 )2、–SO2N(R 6 )2、–SO2R 5A Or optionally substituted alkoxy groups;

[0451] Each R 6 Independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkoxyalkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 3-8 cycloalkyl or optionally substituted C 1-9 heteroaryl; or two R 6Together with the atoms it is attached to, they combine to form optionally substituted C atoms. 2-9 Heterocyclic groups;

[0452] Q is an optional substitution for C. 2-9 Heterocyclic groups, optionally substituted C 3-8 Cycloalkylene, optionally substituted C 1-9 heteroaryl or optionally substituted C 6-10 Alpha-aryl; and

[0453] X is hydrogen or halogen.

[0454] In some embodiments, in compounds of formula (II), (I), or (Ib):

[0455] Each Y is independently either N or CR 4 ;

[0456] R 1 H or C with optional substitution 1-6 alkyl;

[0457] R 2 C is an optional replacement 1-6 Alkyl, optionally substituted C 3-8 cycloalkyl, optionally substituted C 2-9 Heterocyclic groups, optionally substituted C 6-10 aryl, optionally substituted C 1-9 Heteroaryl, optionally substituted C 1-9 heteroaryl C 1-6 Alkyl, –N(R) 5 )2、–CON(R 6 )2、–SO2N(R 6 )2 or –SO2R 5A ;

[0458] R 3 C is an optional replacement 1-9 Mixed aromatics;

[0459] Each R 4 Independently H or optionally substituted C 1-6 alkyl;

[0460] Each R 5 Independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 heteroaryl or –SO2R 5A , where each R 5A C can be substituted independently. 1-6Alkyl or optionally substituted C 3-8 cycloalkyl; or two R 5 Together with the atoms it is attached to, they combine to form optionally substituted C atoms. 2-9 Heterocyclic groups;

[0461] Each R 5A C can be substituted independently. 1-6 Alkyl or optionally substituted C 3-8 cycloalkyl; and

[0462] Each R 6 Independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 aryl or optionally substituted C 1-9 heteroaryl; or two R 6 Together with the atoms it is attached to, they combine to form optionally substituted C atoms. 2-9 Heterocyclic group.

[0463] The compounds of this invention may be, for example, compounds of formula (Ia):

[0464]

[0465] Or a pharmaceutically acceptable salt thereof, wherein Y, R 1 R 2 R 3 and R 4 As described in equation (I).

[0466] The compounds of this invention may be, for example, compounds of formula (Ib):

[0467]

[0468] Or a pharmaceutically acceptable salt thereof, wherein Y, R 1 R 2 R 3 and R 4 As described in equation (I).

[0469] The compounds of this invention may be, for example, compounds of formula (IA):

[0470]

[0471] or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 R 3 and R 4 As described in equation (I).

[0472] Compounds of formula (IA) can be, for example, compounds of formula (IA-a):

[0473]

[0474] or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 R 3 and R 4 As described in equation (I).

[0475] The compounds of this invention may be, for example, compounds of formula (IB):

[0476]

[0477] or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 R 3 and R 4 As described in equation (I).

[0478] Compounds of formula (IB) can be, for example, compounds of formula (IB-a):

[0479]

[0480] or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 R 3 and R 4 As described in equation (I).

[0481] The compounds of this invention may be, for example, compounds of formula (IC):

[0482]

[0483]

[0484] or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 R 3 and R 4 As described in equation (I).

[0485] Compounds of formula (IC) can be, for example, compounds of formula (IC-a):

[0486]

[0487] or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 R 3 and R 4 As described in equation (I).

[0488] The compounds of this invention may be, for example, compounds of formula (ID):

[0489]

[0490] or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 R 3 and R 4 As described in equation (I).

[0491] Compounds of formula (ID) can be, for example, compounds of formula (ID-a):

[0492]

[0493] or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 R 3 and R 4 As described in equation (I).

[0494] Preferably, R 1 It is a methyl group.

[0495] In the compounds of this invention, R 2 C can be, for example, arbitrarily substituted. 3-8 Cycloalkyl groups. For example, R 2 It can be a group of formula (A):

[0496]

[0497] in

[0498] n is 0, 1, 2, or 3; and

[0499] R 7 Hydrogen, alkylsulfonyl, cyano, –CON(R) A )2、-SON(R A 2. Optional substitution of C 1-9 Heteroaryl, hydroxyl, or alkoxy, wherein each R A Independently H or alkyl; or two R A Together with the atoms it is attached to, they combine to form C. 2-9 Heterocyclic group.

[0500] In the compounds of this invention, R 2 C can be, for example, arbitrarily substituted. 1-6 Alkyl groups, for example, optionally substituted tertiary C groups 3-6 Alkyl group. For example, R 2 It can be a group of formula (B):

[0501]

[0502]

[0503] where R 7 Hydrogen, alkylsulfonyl, cyano, –CON(R) A )2、-SON(R A 2. Optional substitution of C 1-9 Heteroaryl, hydroxyl, or alkoxy, wherein each R A Independently H or alkyl; or two R A Together with the atoms it is attached to, they combine to form C. 2-9 Heterocyclic group.

[0504] In the compounds of this invention, R 2 For example, non-aromatic C can be substituted by any choice. 2-9 Heterocyclic group.

[0505] In the compounds of this invention, R 2 For example:

[0506] –I、–SO2Me、 –SO2Ph、 –OMe、 –OCH2CF3、

[0507]

[0508]

[0509] In the compounds of this invention, R 3 It can be, for example, a monocyclic C that has been optionally substituted with at least one nitrogen atom (e.g., two nitrogen atoms). 1-9 heteroaryl. For example, R 3 It can be a group of formula (C):

[0510]

[0511] Where A is an optionally substituted monocyclic C 1-9 Mixed aromatic rings.

[0512] In some compounds of the present invention, A may be, for example, a group of formula (C1):

[0513]

[0514] where R 8 C is hydrogen, halogen, or optionally substituted. 1-6 alkyl.

[0515] In the compounds of this invention, R 3 For example:

[0516]

[0517] In the compounds of this invention, R 3 For example:

[0518]

[0519] In the compounds of this invention, R 4 It can be, for example, hydrogen.

[0520] The compounds of the present invention may be, for example, the compounds listed in Table 1 below or pharmaceutically acceptable salts thereof.

[0521] Table 1

[0522]

[0523]

[0524]

[0525]

[0526]

[0527]

[0528]

[0529]

[0530]

[0531]

[0532] This invention includes (where possible) individual diastereomers, enantiomers, epiomers, and transisomers of the compounds disclosed herein, as well as mixtures of their diastereomers and / or enantiomers, including racemic mixtures. While the specific stereochemistry disclosed herein is preferred, other stereoisomers, including diastereomers, enantiomers, epiomers, transisomers, and mixtures thereof, may also be used to treat ATR-mediated diseases. Inactive or less active diastereomers and enantiomers may be used, for example, in scientific research related to receptors and activation mechanisms.

[0533] It should be understood that some molecules can exist in multiple tautomer forms. Although the examples may indicate only one tautomer, the present invention includes all tautomers.

[0534] The present invention also includes pharmaceutically acceptable salts of the compounds and pharmaceutical compositions comprising the compounds and pharmaceutically acceptable carriers. The compounds are particularly useful for, for example, certain types of cancer and for slowing the progression of cancer when it develops in a patient.

[0535] The compounds disclosed herein can be used in pharmaceutical compositions comprising (a) a compound or a pharmaceutically acceptable salt thereof and (b) a pharmaceutically acceptable carrier. The compounds can be used in pharmaceutical compositions comprising one or more other active pharmaceutical ingredients. The compounds can also be used in pharmaceutical compositions in which the compound disclosed herein or a pharmaceutically acceptable salt thereof is the sole active ingredient.

[0536] Optical isomers - diastereomers - geometric isomers - tautomers

[0537] The compounds disclosed herein may contain, for example, one or more stereoisomeric source centers and may exist as racemates, racemic mixtures, single enantiomers, individual diastereomers, and mixtures of diastereomers and / or enantiomers. This invention includes all such isomeric forms of the compounds disclosed herein. It is intended that all possible stereoisomers (e.g., enantiomers and / or diastereomers) of the compounds, in the form of mixtures and as pure or partially purified compounds, be included within the scope of this invention (i.e., possible combinations of stereoisomeric source centers as pure compounds or in the form of mixtures).

[0538] Some of the compounds described herein may contain rotationally hindered bonds, allowing two separate rotational isomers or trans-isomers to be isolated and found to have potentially advantageous different biological activities. It is intended that all possible trans-isomers be included within the scope of this invention.

[0539] Some of the compounds described herein may contain olefinic double bonds, and unless otherwise specified, this means that E and Z geometric isomers are included.

[0540] Some of the compounds described herein can exist at different hydrogen bonding sites, referred to as tautomers. Examples include ketones and their enol forms, referred to as keto-enol tautomers. Individual tautomers and mixtures thereof are covered by this invention.

[0541] The compounds disclosed herein that have one or more asymmetric centers can be isolated into diastereomers, enantiomers, etc., by methods well known in the art.

[0542] Alternatively, enantiomers and other compounds with chiral centers can be synthesized by stereotactic synthesis using optically pure starting materials and / or reagents with known configurations.

[0543] Metabolites – Prodrugs

[0544] This invention includes therapeutically active metabolites, wherein the metabolites themselves fall within the scope of the claims. This invention also includes prodrugs, which are compounds that are converted into the claimed compound upon or after administration to a patient. In some cases, the claimed chemical structure of this application may itself be a prodrug.

[0545] Isotope-enriched derivatives

[0546] This invention includes molecules enriched with isotopes at one or more sites within the molecule. Therefore, compounds enriched with deuterium fall within the scope of the claims.

[0547] Methods for preparing the compounds of the present invention

[0548] The compounds of the present invention can be prepared using reactions and techniques known in the art, as well as those described herein.

[0549] Method A

[0550] The compounds of the present invention can be prepared as shown in Scheme A and described herein. A commercially available 4-cyano-7-azaindole can be hydrolyzed to an acid and esterified under standard conditions. Regiospecific chlorination at the 6-position can be achieved by 7-aza oxidation with an oxidant (e.g., mCPBA) followed by chlorination with methanesulfonyl chloride. The indole nitrogen can be protected with a suitable protecting group (PG) (e.g., SEM or THP). The compounds can be prepared by S-catalyzed optionally by palladium (O) or copper (I). N Under Ar conditions, the chlorine at the 6-position can be replaced with a suitably substituted morpholine. The ester can then be derivatized by reduction to an alcohol using a suitable reducing agent (e.g., LiBH4 or DIBAL-H), activated by the formation of a methanesulfonate or iodo group, and substituted with sodium methanesulfonate to form a methyl sulfone. Cyclopropanization at the benzyl position can be accomplished using dibromoethane in the presence of a base and a phase-transfer catalyst. Deprotection of the azidoindole then yields a key intermediate, which can be obtained by replacing the chlorine at the 6-position with a suitable aryl iodine or heteroaryl iodine (R... 3 -I) is derivatized by palladium or copper catalytic coupling to generate the compound of the present invention. In R 3 In cases where a protecting group is present to facilitate a substitution reaction, the deprotection step may require the use of acid, base, and / or fluoride conditions to obtain the compounds of the present invention.

[0551] Option A

[0552]

[0553] Method B

[0554] The compounds of this invention can also be prepared as shown in Scheme B and described herein. A commercially available 4-chloro-7-azaindole can be activated to an affinity-substituted form by oxidation of the 7-aza group followed by methylation with dimethyl sulfate. Addition of a suitably substituted morpholine, followed by in-situ elimination of methanol, yields 6-morpholinoazaindole. An aryl or heteroaryl group (R) can then be added via a copper-mediated arylation reaction. 3 Depending on the nature of the heteroaryl group, the protecting group may need to be held in the appropriate position prior to this coupling reaction. The 4-chloro group can be derivatized in various different ways to provide the compounds of the present invention; for example, palladium or copper-mediated coupling can be used to achieve the desired effect in R... 2 Positionally mounted aryl or heteroaryl. Alternatively, if R 2 For substituted amines, then in S N Chloride substitution may occur under Ar conditions or under Buchwald-type coupling conditions. Sulfides can also be used to replace the 4-chloro group, which can optionally be oxidized to generate sulfones. In R... 3 In the case of a protecting group, the deprotection step may require the use of acid, base, and / or fluoride conditions to obtain the compounds of the present invention.

[0555] Option B

[0556]

[0557] Method C

[0558] The compounds of this invention can be prepared from key intermediate A, which can be prepared as shown in Scheme C and described herein. The protected 5-aminopyrazole can be prepared by condensing a suitable aldehyde with hydrazine hydrate and acrylonitrile. Then, condensation with a dialkyl oxaloacetate in refluxed acetic acid yields a substituted azidazole. The hydroxyl group is activated with trifluoromethanesulfonic anhydride, followed by nucleophilic substitution with a morpholine derivative to generate key intermediate A.

[0559] Option C

[0560]

[0561] Method D

[0562] Intermediate A can be converted into the compounds of the present invention by converting an alkyl ester to a group as shown in Scheme D and described herein. For example, treating intermediate A with a reducing group (e.g., DIBAL-H, LiBH4, or NaBH4) yields a primary alcohol, which can be activated with a reagent such as MsCl or TsCl. Replacing the leaving group with an alkyl sulfonate provides benzyl sulfone, which can be alkylated with an alkyl halide under basic conditions. Deprotection and arylation are then performed as described in Method A to obtain the compounds of the present invention.

[0563] Option D

[0564]

[0565] Method E

[0566] The compounds of the present invention can be prepared from intermediate A as shown in Scheme E and described herein. Intermediate A can be treated with an alkylating agent such as magnesium methyl bromide to convert the alkyl ester group into a tertiary alcohol. This substance can be deprotected and arylated as described in Method A to obtain the compounds of the present invention.

[0567] Option E

[0568]

[0569] Method F

[0570] The compounds of the present invention can be prepared from intermediate A as shown in Scheme F and described herein. Intermediate A can be deprotected under acidic conditions and then arylated under copper-catalyzed conditions. The ester group can then be reduced and optionally activated with an agent (e.g., methanesulfonyl chloride or toluenesulfonyl chloride) in the presence of lithium iodide. Substitution with sodium cyanide is then performed to provide arylacetonitrile, an exemplary compound of the present invention. Compounds of this type can be alkylated with an alkyl halide in the presence of a base to provide dialkylated arylacetonitrile, which is a compound of the present invention. If RX is a dihaloalkane, the corresponding cyclic derivative will be formed, wherein the two R groups form a 3-7 membered ring. Alternatively, the primary alcohol can be coupled with a cyanohydrin under Mitsunobu conditions to directly give the nitrile derivative. The nitrile can also be hydrolyzed to a primary amide under basic conditions or in the presence of a metal catalyst to provide the compounds of the present invention.

[0571] Option F

[0572]

[0573] Method G

[0574] The compounds of this invention can be prepared as shown in Scheme G and described herein. N-protected 5,7-dichloro-3H-imidazo[4,5-b]pyridine can be treated with arylboronic acid under palladium catalysis to install a suitable R... 2 Group. In S, which can be optionally catalyzed by palladium (0) or copper (I), N Under Ar conditions, the second chlorine substituent can be replaced with a suitably substituted morpholine. The protecting group is removed as described in Method A, followed by arylation to obtain the compound of this invention.

[0575] Option G

[0576]

[0577] Method H

[0578] The compounds of the present invention can be prepared as shown in Scheme H and as described herein. An alkyl Grignard reagent is added to the protected azirindole as described in Method A to generate a tertiary alcohol. The protecting group on the azirindole is removed as described in Method A, followed by arylation to obtain the compounds of the present invention.

[0579] Option H

[0580]

[0581] Method I

[0582] The compounds of the present invention can be prepared from intermediate A as shown in Scheme I and described herein. The ester of intermediate A can be hydrolyzed to the corresponding acid, followed by amide treatment under amide-forming conditions using a suitable coupling agent (e.g., EDC or HATU). Deprotection of the azaindole then yields the key intermediate, which can be prepared by adding a suitable aryl iodine or heteroaryl iodine (R... 3 -I) is derivatized by palladium or copper catalytic coupling to generate the compound of the present invention. In R 3 In cases where a protecting group is present to facilitate a substitution reaction, the deprotection step may require the use of acid, base, and / or fluoride conditions to obtain the compounds of the present invention.

[0583] Option I

[0584]

[0585] Method J

[0586] The compounds of this invention can be prepared as shown in Scheme J and described herein. Under standard conditions (e.g., aqueous solutions of LiOH or NaOH), the ester intermediate of method F can be hydrolyzed to the corresponding acid. This acid can be coupled with a hydrazine using an activator (e.g., CDI or EDC) to generate an acylhydrazine. This can be formylated (R=H) or acylated (R=alkyl, aryl) to give a diacylhydrazine, which can be cyclized to obtain the compounds of this invention. Cyclization with POCl3 yields an oxadiazole. Cyclization with Lawrsson reagent yields a thiadiazole. 3 In cases where a protecting group is present to facilitate these cyclizations, the deprotection step may require the use of acid, base, and / or fluoride conditions to obtain the compounds of the present invention.

[0587] Scheme J

[0588]

[0589] Method K

[0590] The compounds of the present invention can be prepared as shown in scheme K and described herein. In S, optionally catalyzed by palladium (0) or copper (I), N Under Ar conditions, 5-chloro-3H-[1,2,3]triazolo[4,5-b]pyridine can be replaced with a suitably substituted morpholine. This can be achieved by using a suitable aryl iodine or heteroaryl iodine (R... 3 Palladium or copper catalytic coupling of -I) to derivatize triazole nitrogen to install appropriate R 3 The 7-chloro group can be regiospecifically chlorinated at the 7-position by 3-azaoxidation with an oxidizing agent (e.g., mCPBA) followed by chlorination with methanesulfonyl chloride. The 7-chloro group can be derivatized in various ways to provide the compounds of the present invention; for example, palladium or copper-mediated coupling can be used to achieve regiospecific chlorination at the 7-position. 2 Positionally mounted aryl or heteroaryl. Alternatively, if R 2 For substituted amines, then in S N Chloride substitution may occur under Ar conditions or under Buchwald-type coupling conditions. Sulfides can also be used to replace the 4-chloro group, which can optionally be oxidized to generate sulfones. In R... 3 In cases where a protecting group is present to facilitate these cyclizations, the deprotection step may require the use of acid, base, and / or fluoride conditions to obtain the compounds of the present invention.

[0591] Option K

[0592]

[0593] Method L

[0594] 2,6-Difluoro-4-iodopyridine can be formylated by metallization with a strong base and then captured by a suitable formylating agent (such as ethyl formate). The resulting aldehyde can be condensed with a suitably substituted pyrazolhydrazine to form the corresponding hydrazine, which can then be cyclized to an azainazole by heating to a high temperature. This can be achieved in S... N Under Ar conditions, key intermediate B is provided by replacing the fluorine substituent on azidoinazole with a suitably substituted morpholine. Protecting pyrazole NH with a suitable protecting group yields key intermediate C, typically a mixture of N-protected regioisomers.

[0595] Option L

[0596]

[0597] Method M

[0598] The compounds of the present invention can be prepared as shown in Scheme M and described herein, and the key intermediate C can be treated with arylboronic acid under palladium catalysis to install appropriate R. 2 Protecting groups. The compounds of this invention are obtained by removing the protecting groups.

[0599] Option M

[0600]

[0601] Method N

[0602] The compounds of the present invention can be prepared as shown in Scheme N and described herein. The key intermediate C can be metallized with alkyllithium or alkylmagnesium halide to generate aryllithium or arylmagnesium bromide, which can be added to a suitable ketone to generate a tertiary alcohol derivative. In the case that the ketone contains one or more deuterium-enriched sites, the resulting product will also be deuterium-enriched. Removing the protecting group yields the compounds of the present invention. Alternatively, this chemistry can be carried out using the key intermediate B in the absence of a protecting group to directly provide the compounds of the present invention.

[0603] Option N

[0604]

[0605] Method O

[0606] The compounds of this invention can be prepared as shown in Scheme O and described herein. The key intermediate C can be treated with a carbon, nitrogen, or sulfur-based nucleophile to replace the iodide group and mount a suitable R. 2 Protecting groups. The compounds of this invention are obtained by removing the protecting groups.

[0607] Option O

[0608]

[0609] Method P

[0610] The compounds of the present invention can be prepared as shown in Scheme P and described herein. Treatment with a brominating agent can introduce a bromine atom into the 3-position of the azidazole ring, thereby providing the compounds of the present invention. Treatment with alkyl, vinyl, or aryltinane under Pd catalysis provides the compounds of the present invention.

[0611] Plan P

[0612]

[0613] Method Q

[0614] The compounds of the present invention can be prepared as shown in Scheme Q and as described herein. 2,6-Difluoro-4-iodonicotinaldehyde can be cyclized with hydrazine to form 6-fluoro-4-iodo-1H-pyrazolo[3,4-b]pyridine, which can then be subjected to S-reaction with a substituted morpholine. N Ar reaction. The resulting intermediate can undergo a second S reaction with 2-cyanopropane under alkaline conditions. N Ar reaction was performed to yield a disubstituted azidazole ring system. Ullmann coupling was then carried out on the NH group of the azidazole, followed by deprotection, to provide the compounds of this invention.

[0615] Solution Q

[0616]

[0617] Method R

[0618] The compounds of this invention can be prepared as shown in Scheme R and described herein. Intermediate C can be converted to a borate reagent by treatment with bis(pinacol)diboron, a palladium catalyst, and a base. This borate can then be treated with an aryl halide or trifluoromethanesulfonic acid under palladium catalysis to install a suitable R. 2 Protecting groups. The compounds of this invention are obtained by removing the protecting groups.

[0619] Solution R

[0620]

[0621] Method S

[0622] The compounds of this invention can be prepared as shown in Scheme R and described herein. Intermediate C can be chlorinated at position 5. The resulting intermediate can be treated with arylboronic acid or arylboronic ester under palladium catalysis to mount a suitable R. 2 Protecting groups. The compounds of this invention are obtained by removing the protecting groups.

[0623] Option S

[0624]

[0625] Method T

[0626] The compounds of this invention can be prepared as shown in Scheme T and described herein. 2,6-Difluoro-4-iodopyridin-3-carboxaldehyde can be treated with a substituted morpholine to selectively replace the 6-fluoro substituent. Oxidation of the aldehyde is then carried out, followed by formation of an acylhydrazine with a suitably protected heterocyclic hydrazine. The acylhydrazine can be cyclized under basic conditions to form an iodopyrazolidine pyridinone ring system. This intermediate can undergo S-reaction with carbon, oxygen, or sulfur nucleophiles. N Ar substitution reaction, or preferably treatment with arylboronic acid under palladium catalysis, to install a suitable R 2 Protecting groups are then removed to provide the compounds of the present invention.

[0627] Solution T

[0628]

[0629] Treatment

[0630] The compounds of the present invention can be used to treat ATR kinase-mediated diseases or conditions in subjects by administering an effective amount of the compounds of the present invention to the subject.

[0631] A symptom of a disease or condition can be excessive cell proliferation. For example, a disease or condition can be cancer. Cancer can be, for example, epithelial carcinoma, sarcoma, adenocarcinoma, lymphoma, leukemia, or melanoma. Cancer can also be, for example, a solid tumor.

[0632] Non-limiting examples of cancer include prostate cancer, breast cancer, ovarian cancer, multiple myeloma, brain cancer, glioma, lung cancer, salivary gland cancer, stomach cancer, thymic carcinoma, thyroid cancer, leukemia, melanoma, lymphoma, gastric cancer, pancreatic cancer, kidney cancer, bladder cancer, colon cancer, and liver cancer.

[0633] Non-limiting examples of epithelial carcinoma include thyroid brain-like epithelial carcinoma, familial thyroid brain-like epithelial carcinoma, acinar epithelial carcinoma, acinar epithelial carcinoma, adenocystic epithelial carcinoma, adenoid cystic epithelial carcinoma, adenomatous epithelial carcinoma, adrenocortical epithelial carcinoma, alveolar epithelial carcinoma, alveolar cell epithelial carcinoma, basal cell epithelial carcinoma, basal cell carcinoma, basaloid epithelial carcinoma, basal squamous cell carcinoma, bronchioloalveolar epithelial carcinoma, bronchiolar epithelial carcinoma, bronchial epithelial carcinoma, medullary epithelial carcinoma, bile duct cell epithelial carcinoma, choriocarcinoma, colloid epithelial carcinoma, comedo-like epithelial carcinoma, and the main body. Epithelial carcinoma, cribriform carcinoma, armored carcinoma, cutaneous epithelial carcinoma, columnar carcinoma, columnar cell carcinoma, ductal carcinoma, sclerothelial carcinoma, embryonic carcinoma, cerebral carcinoma, epidermoid carcinoma, adenoid carcinoma, exophytic epithelial carcinoma, ulcerative epithelial carcinoma, fibrous epithelial carcinoma, colloidal carcinoma, gelatinous carcinoma, giant cell carcinoma, giant cell carcinoma, adenocarcinoma, granular cell carcinoma, pilostromal carcinoma, polyangiogenic carcinoma, hepatocellular carcinoma, Schulte's cell carcinoma, hyaline carcinoma, adrenoid carcinoma, infantile embryonic carcinoma. In situ carcinoma, intraepithelial carcinoma, intraepithelial carcinoma, Klinefelter's carcinoma, Kurtschitzky cell carcinoma, large cell carcinoma, lenticular carcinoma, lenticular epithelial carcinoma, lipomatous carcinoma, lymphoepithelial carcinoma, medullary carcinoma, medullary carcinoma, melanoma, soft epithelial carcinoma, mucinous carcinoma, mucinous epithelial carcinoma, mucinous cell carcinoma, mucinous epidermoid carcinoma, mucinous epithelial carcinoma, mucinous epithelial carcinoma, myxomatous carcinoma, nasopharyngeal carcinoma, oat cell carcinoma, ossifying carcinoma, osteoid carcinoma, papillary carcinoma, periportal carcinoma, pre-invasive carcinoma Epithelial carcinoma, acanthosis nigra, nephrotic cell carcinoma, renal cell carcinoma, reservoir cell carcinoma, sarcomatoid carcinoma, Schneider's carcinoma, sclerosing carcinoma, scrotal carcinoma, signet ring cell carcinoma, simple epithelial carcinoma, small cell carcinoma, potato-shaped carcinoma, spherical cell carcinoma, spindle cell carcinoma, cavernous carcinoma, squamous cell carcinoma, string carcinoma, capillary carcinoma, angiotensinus carcinoma, transitional cell carcinoma, nodular carcinoma, nodular epithelial carcinoma, verrucous carcinoma, and villous carcinoma.

[0634] Non-limiting examples of sarcomas include chondrosarcoma, fibrosarcoma, lymphosarcoma, melanosarcoma, myxosarcoma, osteosarcoma, Ebernathy's sarcoma, liposarcoma, liposarcoma, alveolar soft tissue sarcoma, ameloblastic sarcoma, botryoid sarcoma, chloroma sarcoma, choriocarcinoma, embryonal sarcoma, Williams' tumor sarcoma, endometrial sarcoma, stromal sarcoma, Ewing's sarcoma, fascial sarcoma, fibroblastic sarcoma, giant cell sarcoma, granulocytic sarcoma, Hodgkin's sarcoma, idiopathic multiple pigmented hemorrhagic sarcoma, B-cell immunoblastic sarcoma, T-cell immunoblastic sarcoma, Jensen's sarcoma, Kaposi's sarcoma, Kupffer cell sarcoma, angiosarcoma, leukemic sarcoma, malignant mesenchymal sarcoma, paraosteal sarcoma, reticulum cell sarcoma, Rouss's sarcoma, serous cystic sarcoma, synovial sarcoma, and capillary hemangiosarcoma.

[0635] Non-limiting examples of leukemia include acute non-lymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, acute promyelocytic leukemia, adult T-cell leukemia, non-leukemic leukemia, leukemic leukemia, basophilic leukemia, blastic leukemia, bovine leukemia, chronic myeloid leukemia, cutaneous leukemia, embryonic leukemia, eosinophilic leukemia, Gross's leukemia, hairy cell leukemia, hematopoietic leukemia, hematopoietic leukemia, histiocytic leukemia, stem cell leukemia, acute monocytic leukemia, and leukopenic leukemia. Blood diseases, lymphocytic leukemia, lymphoblastic leukemia, lymphocytic leukemia, lymphoid leukemia, lymphosarcoma cell leukemia, mast cell leukemia, megakaryocytic leukemia, micromyeloblastic leukemia, monocytic leukemia, myeloblastic leukemia, myeloid leukemia, myeloid granulocytic leukemia, granulocytic-monocytic leukemia, Neghly leukemia, plasma cell leukemia, multiple myeloma, plasma cell leukemia, promyelocytic leukemia, Riddle cell leukemia, Schilling leukemia, stem cell leukemia, subleukemic leukemia, and undifferentiated cell leukemia.

[0636] Non-limiting examples of melanoma include acral-lentiginous melanoma, amelanoma, benign juvenile melanoma, Claudemann melanoma, S91 melanoma, Harper-Pascal II melanoma, juvenile melanoma, malignant lentiginous melanoma, malignant melanoma, nodular melanoma, subungual melanoma, and superficial diffuse melanoma.

[0637] The compounds of the present invention can be administered via routes selected from the group consisting of: oral, sublingual, buccal, transdermal, intradermal, intramuscular, parenteral, intravenous, intra-arterial, intracranial, subcutaneous, intraorbital, intravenous, intraspinal, intraperitoneal, intranasal, inhalation, intratumoral, and local administration.

[0638] The method of the present invention may include the step of identifying a subject as a candidate for ATR inhibitor therapy. For example, a subject may be identified as a candidate for ATR inhibitor therapy by determining whether: (i) the subject has cancer with a defective ATM signaling cascade; (ii) the subject has cancer, cancer cells, or cells with a genetic abnormality expressing a cancer driver gene or oncogene; (iii) the subject has cancer, cancer cells, or cells with one or more defects in proteins and genes involved in base excision repair; (iv) the subject has cancer with defects in proteins or genes involved in homologous recombination; (v) the subject has cancer with defects in proteins or genes associated with sensitivity to ATR inhibitors or genetic perturbations of ATR; or (vi) the subject has cancer with genetic or protein characteristics associated with sensitivity to ATR inhibitors.

[0639] The described compounds, compositions, and methods may be used to treat subjects with cancers exhibiting abnormalities in the ATM signaling cascade. For example, abnormalities in the ATM signaling cascade may be due to altered expression or activity of one or more of the following proteins / genes, including but not limited to: ATM, p53, CHK2, MRE11, RAD50, NBS1, 53BP1, MDC1, H2AX, MCPH1 / BRIT1, CTIP, and SMC1. Abnormalities in ATM signaling can be identified as follows: changes in CHK2 phosphorylation of 20% or greater may indicate an abnormality in the ATM signaling cascade, or the inability of cells to arrest in the G1 and S phases of the cell cycle in response to double-strand DNA breaks may also indicate an abnormality in the ATM signaling cascade.

[0640] The described compounds, compositions, and methods can be used to treat subjects with cancers, cancer cells, or cells exhibiting abnormal expression of cancer driver protein oncogenes. For example, cancer cells may have genetic abnormalities that result in altered expression or activity of one or more of the following proteins / genes, including but not limited to KRAS, NRAS, HRAS, BRAF, MYC, MOS, E2F, CDC25A, CDC4, CDK2, CCNE1, CCNA1, DNAPK, APOBEC3, CDC6, and RB1.

[0641] The described compounds, compositions, and methods may be used to treat subjects with cancer, cancer cells, or cells that have one or more abnormalities in proteins or genes involved in base excision repair. For example, abnormalities in base excision repair proteins may be alterations in the expression or genes of one or more of the following proteins / genes, including but not limited to UNG, SMUG1, MBD4, TDG, OGG1, MYH, NTH1, MPG, NEIL1, NEIL2, NEIL3 (DNA glycosylases); APE1, APEX2 (AP endonucleases); LIG1, LIG3 (DNA ligases I and III); XRCC1 (LIG3 accessory); PNK, PNKP (polynucleotide kinases and phosphatases); PARP1, PARP2 (poly(ADP-ribose) polymerases); PolB, PolG (polymerases); FEN1 (endonucleases); or Aprataxin.

[0642] The described compounds, compositions, and methods may be used to treat subjects with cancer, cancer cells, or cells that have one or more abnormalities in proteins or genes involved in homologous recombination. For example, the abnormality in homologous recombination may be an alteration in the expression or activity of one or more of the following proteins / genes, including but not limited to: BRCA1, BRCA2, MRE11, RAD50, RAD51, RAD52, RAD54L, NBN, ATM, H2AX, PALB2, RPA, BRIP1, BARD1, ATR, ATRX, CHK1, CHK2, MDM2, MDM4, FANCA, FANCC, FANCD2, FANCE, FANCF, FANCG, and FANCL.

[0643] The described compounds, compositions, and methods may be used to treat subjects with cancer, cancer cells, or cells exhibiting one or more abnormalities in proteins or genes associated with sensitivity to ATR inhibitors or genetic perturbations of the ATR signaling pathway. For example, gene abnormalities associated with sensitivity to ATR inhibitors or genetic perturbations of ATR may be alterations in the expression or activity of one or more of the following proteins / genes, including but not limited to: ATR, CHK1, ERCC1, ERCC2, RAD17, RAD1, RAD9A, ERCC4, ATM, FANCE, GCP3, IDH1, PALB2, PMS2, ARID1A, SLX4, MSH4, RRM2, POLA, POLD1, RRM1, WEE1, CLSPN, PGBD5, XRCC1, XRCC3, XRCC5, KDM5D, CDC6, SLFN11, TLK1, and TLK2.

[0644] Numerous methods are known in the art for determining whether a tumor has protein or gene abnormalities. For example, sequencing of the genomic DNA or mRNA product of each specified gene (e.g., UNG, PARP1, or LIG1) can be performed on a tumor sample to determine the presence of mutations that are expected to regulate the function or expression of the gene product. In addition to mutational inactivation, tumor cells can also regulate genes by methylating promoter regions, leading to decreased gene expression. This is most commonly assessed using methylation-specific polymerase chain reaction (PCR) to quantify the methylation levels of promoters of base excision repair genes of interest. Analysis of DNA repair gene promoter methylation is commercially available.

[0645] Gene expression levels can be assessed by directly quantifying the mRNA and protein product levels of each gene using standard techniques such as quantitative reverse transcriptase-coupled polymerase chain reaction (RT-PCR), RNA-Seq for gene expression, and immunohistochemistry (IHC) for protein expression. Gene amplification or deletion leading to abnormal overexpression or underexpression of proteins can also be measured using FISH (fluorescence in situ hybridization) analysis with probes specific to the gene of interest.

[0646] The methods described above (gene sequence, promoter methylation, and mRNA expression) can also be used to characterize the state of other genes or proteins of interest (e.g., expression or mutation), such as tumor-expressed oncogenes that damage DNA or defects in cellular DNA repair pathways.

[0647] The described compounds, compositions, and methods may be used to treat subjects with cancers whose genetic characteristics are associated with sensitivity to ATR inhibitors. In some embodiments, the genetic characteristics are one or more of the following: cells possessing a telomere elongation replacement mechanism (ALT) characterized by cell transformation or protein expression in the absence of HTERT and / or ATRX mRNA; presence of C-ring or partially double-stranded, circular extrachromosomal telomere repeat sequences (ECTRs); presence of telomeres of varying lengths and positive staining for ALT-associated promyelocytic leukemia (PML) nucleosomes (APBs).

[0648] Several methods exist for identifying ALT signatures in cells. Non-limiting examples of these methods include: measuring HTERT and ATRX expression via Western blotting, immunohistochemistry (IHC), or mRNA expression (qRT-PCR) analysis; measuring the presence of C-loops in PCR analysis; measuring the presence of telomeres of varying lengths via telomere restriction fragment analysis (TRF), which uses the length distribution of terminal restriction fragments to measure the heterogeneous range of telomere lengths in a cell population; and performing staining for the presence of APB using IHC by co-staining with probes targeting telomere DNA and PML proteins.

[0649] Pharmaceutical Composition

[0650] The compounds used in the methods described herein are preferably formulated as pharmaceutical compositions for administration to human subjects in a biocompatible form suitable for in vivo administration. Pharmaceutical compositions typically comprise the compounds described herein and pharmaceutically acceptable excipients. Some pharmaceutical compositions may comprise one or more additional pharmaceutically active agents described herein.

[0651] The compounds described herein may also be used in the form of free bases; as salts, zwitterions, solvates; or as prodrugs or pharmaceutical compositions thereof. All forms are within the scope of this invention. As will be understood by those skilled in the art, the compounds, their salts, zwitterions, solvates, prodrugs, or pharmaceutical compositions may be administered to the patient in various forms, depending on the chosen route of administration. The compounds used in the methods described herein may be administered, for example, orally, parenterally, buccally, sublingually, nasally, rectally, via patch, pump, or transdermally, and pharmaceutical compositions may be formulated accordingly. Parenterally administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary, intrathecal, rectal, and local administration. Parenterally administration may be performed by continuous infusion over a selected time period.

[0652] For human use, the compounds of the present invention can be administered alone or in combination with a pharmaceutical carrier chosen with regard to the intended route of administration and standard pharmaceutical practice. Therefore, the pharmaceutical compositions used according to the present invention can be formulated in a conventional manner using one or more physiologically acceptable carriers comprising excipients and adjuvants, which facilitate the processing of the compounds of the present invention into pharmaceutically acceptable preparations.

[0653] This invention also includes pharmaceutical compositions that may contain one or more pharmaceutically acceptable carriers. In preparing the pharmaceutical compositions of this invention, the active ingredient is typically mixed with an excipient, diluted by the excipient, or sealed within such a carrier, for example, in the form of capsules, sachets, paper, or other containers. When the excipient acts as a diluent, it can be a solid, semi-solid, or liquid material (physiological saline) that serves as a medium, carrier, or mediator of the active ingredient. Therefore, the composition can be in the form of tablets, powders, lozenges, capsules, elixirs, suspensions, emulsions, solutions, syrups, and soft and hard gelatin capsules. As is known in the art, the type of diluent can vary depending on the intended route of administration. The resulting composition may contain additional agents, such as preservatives.

[0654] Excipients or carriers are selected based on the method and route of administration. Suitable drug carriers and essential pharmaceutical ingredients for drug formulations are described in Remington: The Science and Practice of Pharmacy, 21st Edition, Gennaro, ed., Lippincott Williams & Wilkins (2005) and USP / NF (United States Pharmacopeia / National Formulary), which are well-known references in the art. Examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, tragacanth gum, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose. Formulations may additionally include: lubricants, such as talc, magnesium stearate, and mineral oil; humectants; emulsifiers and suspending agents; preservatives, such as methylparaben and propylparaben; sweeteners; and flavoring agents. Other exemplary excipients are described in Handbook of Pharmaceutical Excipients, 6th Edition, Rowe et al., Pharmaceutical Press (2009).

[0655] These pharmaceutical compositions can be manufactured by conventional methods, such as conventional mixing, dissolving, granulation, pilling, grinding, emulsification, encapsulation, embedding, or lyophilization processes. Methods well-known in the art for the manufacture of formulations can be found, for example, in Remington: The Science and Practice of Pharmacy, 21st edition, edited by Gennaro, Lippincott Williams & Wilkins (2005) and Encyclopedia of Pharmaceutical Technology, edited by J. Swarbrick and J.C. Boylan, 1988–1999, Marcel Dekker, New York. The correct formulation depends on the chosen route of administration. Formulations and preparations of such compositions are well-known to those skilled in the art of pharmaceutical formulation. In preparing formulations, the active compound may be ground to provide an appropriate particle size before being combined with other ingredients. If the active compound is substantially insoluble, it may be ground to a particle size of less than 200 mesh. If the active compound is substantially soluble in water, the particle size may be adjusted by grinding to provide a substantially uniform distribution in the formulation, for example, about 40 mesh.

[0656] dose

[0657] The dosage of the compound used in the methods described herein, or a pharmaceutically acceptable salt or prodrug thereof, or a pharmaceutical composition thereof, may vary depending on a number of factors, such as: the pharmacodynamic properties of the compound; the route of administration; the recipient's age, health, and weight; the nature and severity of symptoms; the frequency of treatment and the type of concurrent treatment (if any); and the clearance rate of the compound in the treated animal. Those skilled in the art can determine an appropriate dosage based on the above factors. The compound used in the methods described herein may initially be administered at a suitable dosage, which may be adjusted as needed based on clinical response. Generally, a suitable daily dose of the compound of the present invention will be the amount of the minimum dose at which the compound effectively produces a therapeutic effect. This effective dose will typically depend on the factors described above.

[0658] The compounds of this invention can be administered to a patient in a single dose or in multiple doses. When multiple doses are administered, the doses can be spaced out, for example, every 1-24 hours, 1-7 days, 1-4 weeks, or 1-12 months. The compounds can be administered according to a schedule or without a predetermined schedule. For example, the active compound can be administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 times daily; every 2 days, every 3 days, every 4 days, every 5 days, or every 6 days; every week, 1, 2, 3, 4, 5, 6, or 7 times; every month, 1, 2, 3, 4, 5, or 6 times; or every year, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 times. It should be understood that for any given subject, the specific dosage regimen should be adjusted over time according to individual needs and the professional judgment of the person administering or supervising the administration of the composition.

[0659] Although the attending physician ultimately determines the appropriate dosage and dosing regimen, the effective amount of the compounds of the present invention can be, for example, any compound described herein with a total daily dose, for example, between 0.05 mg and 3000 mg. Alternatively, the dosage can be calculated using the patient's weight. Such dosage ranges can include, for example, between 10 and 1000 mg (e.g., 50-800 mg). In some embodiments, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or 1000 mg of the compound is administered.

[0660] In the method of this invention, the time period for administering multiple doses of the compound of the invention to a patient can vary. For example, in some embodiments, the dose of the compound of the invention is administered to the patient over a time period of 1-7 days, 1-12 weeks, or 1-3 months. In some embodiments, the compound is administered to the patient over a time period of, for example, 4-11 months or 1-30 years. In some embodiments, the compound is administered to the patient at the onset of symptoms. In any of these embodiments, the amount of the compound administered can vary over the time period of administration. When the compound is administered daily, it can be administered, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 times daily.

[0661] preparation

[0662] Compounds identified as capable of treating any of the conditions described herein, using any of the methods described herein, may be administered to a patient or animal in a unit dosage form, together with a pharmaceutically acceptable diluent, carrier, or excipient. Compounds for this therapy may be produced and isolated using any standard techniques known in the field of medicinal chemistry. Suitable formulations or compositions may be provided using routine pharmaceutical practices to administer the identified compound to a patient suffering from the disease or condition. Administration may be initiated before the patient experiences symptoms.

[0663] Exemplary routes of administration for the compounds used in this invention (e.g., the compounds of this invention) or pharmaceutical compositions thereof include oral, sublingual, buccal, transdermal, intradermal, intramuscular, parenteral, intravenous, intraarterial, intracranial, subcutaneous, intraorbital, intravenous, intraspinal, intraperitoneal, intranasal, inhalation, and topical administration. The compounds are ideally administered with a pharmaceutically acceptable carrier. Formulations of pharmaceutical preparations of the compounds described herein for the treatment of the conditions described herein are also part of this invention.

[0664] Formulations for oral administration

[0665] The pharmaceutical compositions contemplated by this invention include those formulated for oral administration (“oral dosage forms”). Oral dosage forms may be, for example, tablets, capsules, liquid solutions or suspensions, powders, liquids or solid crystals, containing an active ingredient mixed with a non-toxic, pharmaceutically acceptable excipient. These excipients may be, for example, inert diluents or fillers (e.g., sucrose, sorbitol, sugar, mannitol, microcrystalline cellulose, starch (including potato starch), calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate, or sodium phosphate); granulating agents and disintegrants (e.g., cellulose derivatives (including microcrystalline cellulose), starch (including potato starch), croscarmellose sodium, alginate, or alginic acid); binders (e.g., sucrose, glucose, sorbitol, gum arabic, alginate, sodium alginate, gelatin, starch, pregelatinized starch, microcrystalline cellulose, magnesium aluminum silicate, sodium carboxymethyl cellulose, methylcellulose, hydroxypropyl methylcellulose, ethylcellulose, polyvinylpyrrolidone, or polyethylene glycol); and lubricants, flow aids, and anti-sticking agents (e.g., magnesium stearate, zinc stearate, stearic acid, silica, hydrogenated vegetable oil, or talc). Other pharmaceutically acceptable excipients may include colorants, flavoring agents, plasticizers, humectants, buffers, etc.

[0666] Formulations for oral administration may also be presented as chewable tablets, hard gelatin capsules (in which the active ingredient is mixed with an inert solid diluent (e.g., potato starch, lactose, microcrystalline cellulose, calcium carbonate, calcium phosphate, or kaolin)) or soft gelatin capsules (in which the active ingredient is mixed with an aqueous or oil medium (e.g., peanut oil, liquid paraffin, or olive oil)). Powders, granules, and pills may be prepared using the ingredients mentioned above under tablets and capsules, in a conventional manner using, for example, mixers, fluidized bed equipment, or spray drying equipment.

[0667] Controlled-release formulations for oral use can be constructed to release the active drug substance by controlling the dissolution and / or diffusion of the active pharmaceutical ingredient. Any of several strategies can be employed to obtain controlled-release and target plasma concentration curves versus time. In one example, controlled release is achieved by appropriately selecting various formulation parameters and components, including, for example, various types of controlled-release compositions and coatings. Examples include single or multiple unit tablet or capsule compositions, oil solutions, suspensions, emulsions, microcapsules, microspheres, nanoparticles, patches, and liposomes. In some embodiments, the composition comprises a biodegradable, pH- and / or temperature-sensitive polymer coating.

[0668] Controlled release through dissolution or diffusion can be achieved by tablet, capsule, pill, or granule formulations of a suitable coated compound or by incorporating the compound into a suitable matrix. Controlled release coatings may contain coating substances mentioned above and / or, for example, shellac, beeswax, sugar wax, castor wax, carnauba wax, stearyl alcohol, glyceryl monostearate, glyceryl distearate, glyceryl palmitate, ethyl cellulose, acrylic resins, dl-polylactic acid, cellulose acetate butyrate, polyvinyl chloride, polyvinyl acetate, vinylpyrrolidone, polyethylene, polymethacrylate, methyl methacrylate, 2-hydroxymethacrylate, methacrylate hydrogel, 1,3-butanediol, ethylene glycol methacrylate, and / or polyethylene glycol. In controlled release matrix formulations, matrix materials may also include, for example, hydrated methyl cellulose, carnauba wax and stearyl alcohol, carbomer 934, silicone, glyceryl tristearate, methyl acrylate-methyl methacrylate, polyvinyl chloride, polyethylene, and / or halogenated fluorocarbons.

[0669] Compounds and compositions of the present invention may be incorporated into liquid forms for oral administration, including aqueous solutions, suitably flavored syrups, aqueous or oil suspensions, emulsions flavored with edible oils (e.g., cottonseed oil, sesame oil, coconut oil, or peanut oil), elixirs, and similar pharmaceutical carriers.

[0670] Preparations for parenteral administration

[0671] The compounds described herein for use in the methods of this invention can be administered as pharmaceutically acceptable parenteral (e.g., intravenous or intramuscular) formulations as described herein. Pharmaceutical formulations may also be administered parenterally (intravenous, intramuscular, subcutaneously, etc.) in dosage forms or formulations containing conventionally non-toxic, pharmaceutically acceptable carriers and adjuvants. Specifically, formulations suitable for parenteral administration include: aqueous and non-aqueous sterile injectable solutions that may contain antioxidants, buffers, antibacterial agents, and solutes that make the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions that may contain suspending agents and thickeners. For example, to prepare such compositions, the compounds of this invention can be dissolved or suspended in parenteral-acceptable liquid media. Acceptable media and solvents that may be used are water (adjusted to a suitable pH by adding appropriate amounts of hydrochloric acid, sodium hydroxide, or a suitable buffer), 1,3-butanediol, Ringer's solution, and isotonic sodium chloride solution. Aqueous formulations may also contain one or more preservatives, such as methylparaben, ethylparaben, or n-propylparaben. Further information on parenteral formulations can be found, for example, in the United States Pharmacopeia-National Formulary (USP-NF), which is incorporated herein by reference.

[0672] Parenteral preparations may be any one of the five general types of preparations identified by USP-NF as suitable for parenteral administration:

[0673] (1) "Drug injection": a liquid preparation of an active pharmaceutical ingredient (e.g., the compound of the present invention) or a solution thereof;

[0674] (2) "Injection drug": a raw material drug (e.g., the compound of the present invention) in the form of a dry solid, which will be combined with a suitable sterile medium for parenteral administration in the form of a drug injection solution;

[0675] (3) "Drug injection emulsion": a liquid preparation of an active pharmaceutical ingredient (e.g., the compound of the present invention) dissolved or dispersed in a suitable emulsion medium;

[0676] (4) "Drug injection suspension": a liquid preparation of an active pharmaceutical ingredient (e.g., the compound of the present invention) suspended in a suitable liquid medium; and

[0677] (5) "Injectable suspension": a raw material (e.g., the compound of the present invention) in the form of a dry solid, which will be combined with a suitable sterile medium for parenteral administration in the form of an injectable suspension.

[0678] Exemplary parenteral formulations comprise solutions of compounds prepared in water appropriately mixed with a surfactant (e.g., hydroxypropyl cellulose). Dispersions can also be prepared in glycerol, liquid polyethylene glycol, DMSO, and mixtures thereof with or without alcohol, as well as in oils. Under normal storage and use conditions, these preparations may contain preservatives to prevent microbial growth. Routine procedures and ingredients for selecting and preparing suitable formulations are described, for example, in Remington: The Science and Practice of Pharmacy, 21st edition, edited by Gennaro, Lippincott Williams & Wilkins (2005) and United States Pharmacopeia: The National Formulary (USP36NF31), published in 2013.

[0679] Parenteral formulations may contain, for example, excipients, sterile water or saline, polyalkylene glycols (e.g., polyethylene glycol), plant-derived oils, or hydrogenated naphthalene. The release of the compound can be controlled using biocompatible, biodegradable lactide polymers, lactide / glycolic acid copolymers, or polyoxyethylene-polyoxypropylene copolymers. Other potentially available parenteral delivery systems for the compound include ethylene-vinyl acetate copolymer particles, osmotic pumps, implantable infusion systems, and liposomes. Inhalation formulations may contain excipients (e.g., lactose) or may be aqueous solutions containing, for example, polyoxyethylene-9-lauryl ether, heparin salts, and deoxycholates, or may be oily solutions administered as nasal drops or gels.

[0680] Parenteral formulations can be formulated for the rapid release or sustained / prolonged release of compounds. Exemplary formulations for parenteral release of compounds include: aqueous solutions, reconstitution powders, cosolvent solutions, oil / water emulsions, suspensions, oil-based solutions, liposomes, microspheres, and polymer gels.

[0681] combination

[0682] The compounds of the present invention can be administered to a subject in combination with one or more other agents, such as:

[0683] (a) Cytotoxic agents;

[0684] (b) Antimetabolites;

[0685] (c) Alkylating agents;

[0686] (d) Anthracyclines;

[0687] (e) Antibiotics;

[0688] (f) Antimitotic agents;

[0689] (g) Hormone therapy;

[0690] (h) Signal transduction inhibitors;

[0691] (i) Gene expression regulators;

[0692] (j) Apoptosis inducers;

[0693] (k) Angiogenesis inhibitors;

[0694] (l) Immunotherapy agents;

[0695] (m) DNA damage repair inhibitors;

[0696] or

[0697] Its combination.

[0698] Cytotoxic agents can include, for example, actinomycin D, alemtuzumab, alitretinoin, allopurinol, hexamethylmelamine, amifostine, amphotericin B, acridine lactate, arsenic trioxide, asparaginase, azacitidine, azathioprine, and BCG. Calmette-Guérin (BCG), bendamustine, bexarotene, bevacuzimab, bleomycin, bortezomib, busulfan, capecitabine, carboplatin, carfilzomib, carmustine, cetuximab, cisplatin, chlorambucil, cladribine, clofarabine, colchicine, cristaspase, cyclophosphamide, cyclosporine, cytarabine, pinocembrin B, dacarbazine, dextrin, darbepoetinalfa, dasatinib, daunorubicin, 1-dehydrotestosterone, denileukin, dexamethasone, dexrazoxin, dihydroxy anthracin dione, disulfiram, docetaxel, doxorubicin, emirtin, epirubicin, erlotinib, epigallocatechin gallate, epioetinAlfa), Estrogenus estradiol, Ethidium bromide, Etoposide, Everolimus, Filagristin, Finasunate, Fluoxuridine, Fludarabine, 5-FU, Fulvestrant, Ganciclovir, Gerdemycin, Gemcitabine, Glucocorticoids, Bacitracin D, Histamine Relin, Hydroxyurea, Ibrimomab, Idarubicin, Ifosfamide, Imatinib, Irinotecan, Interferon, Interferon α-2a, Interferon α-2b, Ixabepilone, Lactate dehydrogenase A (LDH-A), Lenalidomide, Letrozole, Leucovorin, Levamisole, Lidocaine, Lomustine, Nitrogen mustard, Melphalan, 6-Mercaptopurine, Mestizoline Sodium, methotrexate, methoxsalen, metoprine, metronidazole, mithramycin, mitomycin C, mitoxantrone, nandrolone, nelarabine, nilotinib, nofetumomab, oprelvekin, oxaliplatin, paclitaxel, pemetrexed, pentostatin, palifermin, pamidronate, pegastrobin, pemetrexed disodium, purcamycin, porphyrin sodium, procaine, procarbazine, propranolol, puromycin, quinacrine, rhizobacterium, radioisotope, raltitrexed, rapamycin, rasburicase, salinosporamide A A) Saxaglastine, sunitinib, temozolomide, teniposide, tetracaine, 6-thioguanine, thiotepa, topotecan, toremifene, trastuzumab, tresuzumab, retinoic acid, valrubicin, vincristine, vinblastine, vindesin, vinorelbine, zoledronate, or combinations thereof.

[0699] Antimetabolites may be, for example, methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil dacarbazine, cladribine, pemetrexed, gemcitabine, capecitabine, hydroxyurea, mercaptopurine, fludarabine, pralatrexate, clofarabine, cytarabine, decitabine, fluorouridine, nerabine, trimethotraxate, thioguanine, pentostatin, or combinations thereof.

[0700] Alkylating agents may be, for example, nitrogen mustard, thiotepa, chlorambucil, melphalan, carmustine (BSNU), lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, cis-dichlorodiamine platinum(II) (DDP), cisplatin, hexamethylmelamine, cyclophosphamide, ifosfamide, hexamethylmelamine, hexamethylmelamine, procarbazine, dacarbazine, temozolomide, streptozotocin, carboplatin, cisplatin, oxaliplatin, uramustine, bendamustine, trabectedin, semustine, or combinations thereof.

[0701] Anthracyclines may be, for example, daunorubicin, doxorubicin, azorubicin, adoxorubicin, amrubicin, annamycin, carrubicin, epirubicin, idarubicin, mitoxantrone, valrubicin, or combinations thereof.

[0702] Antibiotics may include, for example, bleomycin, bleomycin, ampicillin (AMC), ampicillin, bamicillin, carbenicillin, cloxacillin, dicloxacillin, flucloxacillin, mezlocillin, nafcillin, oxacillin, piperacillin, pimecrolimus, pimecrolimus, ticarcillin, aztreonam, imipenem, doripenem, ertapenem, meropenem, cephalosporins, clarithromycin, erythromycin, roxithromycin, telithromycin, lincomycin, punamycin, quinupristin, amikacin, gentamicin, kanamycin, neomycin, netilmicin, paromomycin, tobramycin, streptomycin, sulfamethoxazole, and sulfamethoxazole. Oxazole, sulfamethoxazole, demeclocycline, minocycline, oxytetracycline, tetracycline, penicillin, amoxicillin, cephalexin, erythromycin, clarithromycin, azithromycin, ciprofloxacin, levofloxacin, ofloxacin, doxycycline, clindamycin, metronidazole, tigecycline, chloramphenicol, metronidazole, tinidazole, nitrofurantoin, vancomycin, teicoplanin, telavancin, linezolid, cycloserine, rifamycin, polymyxin B, bacitracin, purpuricin, capreomycin, quinolones, daunorubicin, doxorubicin, 4'-deoxydoxorubicin, epirubicin, idarubicin, procamycin, mitomycin C, mitoxantrone, or combinations thereof.

[0703] Antimitotic agents may be, for example, vincristine, vinorelbine, docetaxel, estradiol, ixapril, paclitaxel, maytansinoid, dolastatin, cryptophycin, or combinations thereof.

[0704] Signal transduction inhibitors can include, for example, imatinib, trastuzumab, erlotinib, sorafenib, sunitinib, temsirolimus, vemurafenib, lapatinib, bortezomib, cetuximab, panitumumab, matuzumab, gefitinib, and STI. 571. Rapamycin, Flavopiridol, Imatinib Mesylate, Vatalanib, Semaxinib, Motesanib, Axitinib, Afatinib, Bosutinib, Crizotinib, Cabozantinib, Dasatinib, Entrectinib, Pazopanib, Lapatinib, Vandetanib, or combinations thereof.

[0705] Gene expression regulators may be, for example, siRNA, shRNA, antisense oligonucleotides, HDAC inhibitors, or combinations thereof. HDAC inhibitors may be, for example, trapoxin A, trapoxin B, valproic acid, vorinostat, belinostat, LAQ824, panobinostat, entinostat, tacedinaline, mocetionstat, givinostat, resminostat, abexinostat, quisinostat, rocilinostat, practinostat, CHR-3996, butyric acid, phenylbutyric acid, 4SC202, romidepsin, sirtinol, cambinol, EX-527, nicotinamide, or combinations thereof. Antisense oligonucleotides may be, for example, custirsen, apatorsen, AZD9150, trabadersen, EZN-2968, LErafAON-ETU, or combinations thereof. siRNAs may be, for example, ALN-VSP, CALAA-01, Atu-027, SPC2996, or combinations thereof.

[0706] Hormone therapy may include, for example, luteinizing hormone-releasing hormone (LHRH) antagonists. Examples of hormone therapies include firmagon, leuprorelin, goserelin, buserelin, flutamide, bicalutamide, ketoconazole, aminoglutethimide, prednisone, hydroxyprogesterone caproate, medroxyprogesterone acetate, megestrol acetate, diethylstilbestrol, ethinylestradiol, tamoxifen, testosterone propionate, flumethasone, flutamide, raloxifene, droloxifene, indoxifene (iodoxyfene), 4-hydroxytamoxifen, trivoxifene, keoxifene, LY117018, onanasone, and toremifene citrate. citrate), medroxyprogesterone acetate, exemestane, fazodazole, voroxycycline, letrozole, anastrozole, nilumet, tripterelin, histamine, abiraterone, medroxyprogesterone acetate, diethylstilbestrol, premarin, fluorometholone, retinoic acid, fenvitamin A, troxacitabine, or combinations thereof.

[0707] Apoptosis inducers may be, for example, recombinant human TNF-associated apoptosis-inducing ligand (TRAIL), camptothecin, bortezomib, etoposide, tamoxifen, or combinations thereof.

[0708] Angiogenesis inhibitors may be, for example, sorafenib, sunitinib, pazopanib, everolimus, or combinations thereof.

[0709] Immunotherapy agents may include, for example, monoclonal antibodies, cancer vaccines (e.g., dendritic cell (DC) vaccines), oncolytic viruses, cytokines, adoptive T-cell therapy, BCG, GM-CSF, thalidomide, lenalidomide, pomalidomide, imiquimod, or combinations thereof. Monoclonal antibodies may include, for example, anti-CTLA4, anti-PD1, anti-PD-L1, anti-LAG3, anti-KIR, or combinations thereof. Monoclonal antibodies may be, for example, alenumab, trastuzumab, ibritumomab tiuxetan, brentuximab vedotin, trastuzumab, adorotractuzumab-metanesin, blinatumomab, bevacizumab, cetuximab, pertuzumab, panitumab, ramucirumab, obinutuzumab, ofatumumab, rituximab, pertuzumab, tositumomab, gemtuzumab ozogamicin, tositumomab, or combinations thereof. Cancer vaccines may include, for example, Sipuleucel-T, BioVaxID, NeuVax, DCVax, and SuVaxM. HSP110 chaperone protein combination vaccine, CDX-1401, MIS416, CDX-110, GVAX Pancreas, HyperAcute TM Pancreas, GTOP-99 or Imprime Oncolytic viruses may be, for example, talimogene laherparepvec. Cytokines may be, for example, IL-2, IFNα, or combinations thereof. Adoptive T-cell therapy may be, for example, tisagenlecleucel, axicabtagene ciloleucel, or combinations thereof.

[0710] DNA damage repair inhibitors may be, for example, PARP inhibitors, cell checkpoint kinase inhibitors, or combinations thereof. PARP inhibitors may be, for example, olaparib, rucaparib, veliparib (ABT-888), niraparib (ZL-2306), iniparib (BSI-201), talazoparib (BMN 673), 2X-121, CEP-9722, KU-0059436 (AZD2281), PF-01367338, or combinations thereof. Cell checkpoint kinase inhibitors may be, for example, MK-1775 or AZD1775, AZD7762, LY2606368, PF-0477736, AZD0156, GDC-0575, ARRY-575, CCT245737, PNT-737 or combinations thereof.

[0711] Example

[0712] The following examples are intended to illustrate the present invention. These examples are not intended to limit the invention in any way.

[0713] Example 1. Preparation of the compound

[0714] Compound 1

[0715] Step 1. Purge the suspension of 4-chloro-7-azaindole (25 g) in DMA (140 mL) with vacuum / N2 gas (3 cycles). Then add zinc powder (1.07 g), zinc cyanide (11.26 g), dppf (2.72 g), and Pd2(dba)3 (2.39 g). Purge the mixture again with vacuum / N2 gas (3 cycles) and heat to 120 °C for 4 h. Cool the reaction mixture to 100 °C and add water (428 mL) over 30 min. Then cool the mixture to rt over 2 h. Filter the crude product and wash with water (2 × 95 mL), then add it to 3N HCl (150 mL) and stir the mixture at rt for 2 h. Remove insoluble matter by filtration. Add 50% NaOH aqueous solution to the filtrate until pH 12 is reached. The mixture was filtered and dried to obtain 1H-pyrrolo[2,3-b]pyridine-4-carboxylonitrile (11.6 g), which was a brown solid.

[0716] Step 2. A mixture of 1H-pyrrolo[2,3-b]pyridine-4-carboxynitrile (10.4 g) and NaOH (29 g) in water (100 mL) and EtOH (100 mL) was heated to reflux for 18 h. After cooling to rt, the mixture was treated with concentrated hydrochloric acid until the pH was approximately 2. The solid was collected by filtration and dried under high vacuum to give 1H-pyrrolo[2,3-b]pyridine-4-carboxylic acid (11.8 g) as a brown solid.

[0717] Step 3. At 0 °C, thionyl chloride (12.4 mL) was added dropwise to EtOH (120 mL), and the mixture was stirred at rt for 30 min. Then, 1H-pyrrolo[2,3-b]pyridine-4-carboxylic acid (12.0 g) was added, and the reaction mixture was heated to reflux for 8 h. After cooling to rt, the solvent was removed under reduced pressure. The resulting residue was suspended in water (150 mL), and the pH was adjusted to pH 9 with a saturated aqueous solution of K2CO3. The mixture was extracted with EtOAc (2 × 150 mL). The combined extracts were washed with brine, dried over MgSO4, filtered, and concentrated to dryness to give 10.5 g of 1H-pyrrolo[2,3-b]pyridine-4-carboxylic acid as a brown solid.

[0718] Step 4. At 0 °C, mCPBA (15.5 g) was added in portions to a mixture of 1H-pyrrolo[2,3-b]pyridine-4-carboxylate (9.5 g) and EtOAc (95 mL). The reaction mixture was heated to rt and stirred for 3 h. The precipitate was filtered, washed with EtOAc (3 × 30 mL), and the residue was dried under high vacuum to give 8.8 g of 4-(ethoxycarbonyl)-1H-pyrrolo[2,3-b]pyridine 7-oxide as a pale yellow solid.

[0719] Step 5. To a solution of 25.5 g of 4-(ethoxycarbonyl)-1H-pyrrolo[2,3-b]pyridine 7-oxide in DMF (250 mL), methanesulfonyl chloride (11.5 mL) was added dropwise. The mixture was then heated to 80 °C for 1 h, then cooled to rt, and another 11.5 mL of methanesulfonyl chloride was added. The mixture was heated again at 80 °C for 1 h. After cooling to 0 °C, the reaction mixture was poured into ice water (480 mL) with vigorous stirring. The mixture was then stirred at 0 °C for 2 h. The precipitate was filtered and washed with water (3 × 200 mL). The residue was dried under high vacuum to give 25.0 g of ethyl 6-chloro-1H-pyrrolo[2,3-b]pyridine-4-carboxylate as a beige solid, which was used in subsequent steps without further purification.

[0720] Step 6. At 0°C, NaH (6.68 g) was added to a solution of 25 g of 6-chloro-1H-pyrrolo[2,3-b]pyridine-4-carboxylate in DMF (250 mL) over 45 min, and the mixture was stirred at 0°C for 1 h. SEM-Cl (23.6 mL) was added over 20 min, and the mixture was stirred at 0°C for 1 h. Water (300 mL) was slowly added and the mixture was extracted with EtOAc (2 × 200 mL), washed with brine, dried over MgSO4, filtered, and concentrated to dryness. The residue was purified by silica gel rapid chromatography (15-30% EtOAc / hexane) to give 32.4 g of 6-chloro-1-(((2-trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-4-carboxylate as an orange oil.

[0721] Step 7. To a solution of ethyl 6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-4-carboxylate (32.3 g) in toluene (150 mL), add (R)-3-methylmorpholine (12.4 mL), BINAP (3.4 g), and cesium carbonate (89 g). Degas the mixture (vacuum / argon, 3 cycles) and add palladium acetate (1.0 g), and degas the reaction mixture again, then heat to 120 °C for 4 hours. After cooling to rt, dilute the mixture with EtOAc (500 mL), filter through a diatomaceous earth pad, and wash with EtOAc (2 × 250 mL). The filtrate was concentrated to dryness under reduced pressure and purified by silica gel chromatography (0-40% EtOAc / hexane) to obtain (R)-6-(3-methylmorpholino)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-4-carboxylic acid ethyl ester (26 g), which was a yellow oil.

[0722] Step 8. Add MeOH (0.048 mL) to a solution of (R)-6-(3-methylmorpholino)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-4-carboxylic acid ethyl ester (4.9 g) in THF (80 mL). Heat the reaction mixture to 65 °C, then add 2 M LiBH4 in THF (9 mL) dropwise over 1 h. Stir the reaction mixture at 65 °C for 18 h. After cooling to rt, add acetone (2 mL) and stir at rt for 30 min. Dilute the mixture with a 1:1 saturated NH4Cl aqueous solution / water (100 mL) and extract with EtOAc (2 × 100 mL). Wash the combined organic extracts with brine, dry over MgSO4, filter, and concentrate to dryness. The residue was purified by silica gel chromatography (5-50% EtOAc / hexane) to give (R)-(6-(3-methylmorpholino)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)methanol (3.9 g), which was a yellow gel.

[0723] Step 9. At 0°C, triethylamine (2.8 mL) was added to a solution of (R)-(6-(3-methylmorpholino)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)methanol (7.5 g) in dichloromethane (70 mL), followed by methanesulfonyl chloride (1.55 mL). The reaction mixture was stirred at rt for 90 min and then diluted with dichloromethane (100 mL) and water (100 mL). The layers were partitioned, and the aqueous layer was extracted with dichloromethane (100 mL). The combined organic extracts were washed with brine, dried over MgSO4, filtered, and concentrated to dryness to obtain (R)-(6-(3-methylmorpholino)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)methylmethanesulfonate (9 g), which was a yellow gel and used in subsequent steps without further purification.

[0724] Step 10. LiI (5.3 g) was added to a solution of (R)-(6-(3-methylmorpholino)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)methylmethanesulfonate (9 g) in dioxane (80 mL). The mixture was heated to 100 °C under argon for 2.5 h. After cooling to rt, the mixture was diluted with EtOAc (100 mL) and water (100 mL). The layers were partitioned, and the aqueous layer was extracted with EtOAc (80 mL). The combined organic extracts were washed with 2M sodium bisulfite (80 mL), water (80 mL), and brine (80 mL), dried over MgSO4, filtered, and concentrated to give (R)-(4-(4-iodomethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-methylmorpholine (9.6 g), which was used in subsequent steps without further purification.

[0725] Step 11. Add sodium methanesulfonate (2.4 g) to a solution of (R)-4-(4-iodomethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-methylmorpholine (9.6 g) in DMF (80 mL). Stir the reaction mixture at rt for 18 h. The reaction mixture consisted of EtOAc (100 mL) and water (100 mL). Distribute the layers and extract the aqueous layer with EtOAc (80 mL). Wash the combined organic extracts with an aqueous solution of sodium thiosulfate (80 mL), water (80 mL), and brine, dry to MgSO4, filter, and concentrate. The residue was purified by silica gel chromatography (10-90% EtOAc / hexane) to obtain (R)-3-methyl-4-(4-((methylsulfonyl)methyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)morpholine (7.5 g), which was a gray-green gel.

[0726] Step 12. To a solution of (R)-3-methyl-4-(4-((methanesulfonyl)methyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)morpholine (7.5 g) in toluene (80 mL), add TBAB (1 g) and 50% NaOH (36 mL), followed by 1,2-dibromoethane (2 mL). Heat the mixture to 65 °C for 18 h. Then, while stirring the mixture at 65 °C, add an additional 1,2-dibromoethane (16 mL) via a syringe pump over 18 h. Age the reaction mixture at 65 °C for another 18 h, then cool to rt. Dilute the reaction mixture with EtOAc (200 mL) and water (150 mL), partition the layers, and extract the aqueous layer with EtOAc (100 mL). The combined organic extracts were washed with brine, dried over MgSO4, and concentrated to dryness under reduced pressure. The residue was purified by silica gel chromatography (10-80% EtOAc / hexane) to give (R)-3-methyl-4-(4-(1-(methanesulfonyl)cyclopropyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)morpholine (5.4 g), which was a yellow foam.

[0727] Step 13. At 0 °C, add TFA (18 mL) to a solution of (R)-3-methyl-4-(4-(1-(methanesulfonyl)cyclopropyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)morpholine (5.4 g) in dichloromethane (50 mL). Warm the reaction mixture to rt and stir for 18 h. Add toluene (40 mL) and concentrate the mixture. Dilute the residue with dioxane (40 mL) and adjust the pH of the mixture to pH 10 by adding 3N NaOH. Heat the mixture to 80 °C for 3 h, then cool to rt. Dilute the mixture with EtOAc (150 mL) and water (150 mL). Distribute the layers and extract the aqueous layer with EtOAc (100 mL). Wash the combined organic extracts with brine, dry over MgSO4, filter and concentrate to dryness. The residue was purified by silica gel chromatography (30-100% EtOAc / hexane) to obtain (R)-3-methyl-4-(4-(1-(methanesulfonyl)cyclopropyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)morpholine (1.65 g), which was a pale yellow foam.

[0728] Step 14. At 0°C, cesium carbonate (9.43 g) was added to a solution of 2.5 g of 3-iodo-1H-pyrazole in DMF (25 mL). Then, SEM-Cl (2.8 mL) was added over 15 min. The mixture was stirred at rt for 18 h. Water (60 mL) was slowly added and the mixture was partitioned with Et2O (60 mL). The aqueous layer was extracted with Et2O (30 mL), and the combined organic extracts were washed with water (3 × 50 mL) and brine, dried over MgSO4, filtered, and concentrated to dryness. The residue was purified by silica gel chromatography (0–30% EtOAc / hexane) to give 3-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole (3.3 g) as a colorless liquid. ¹H-NMR showed a ratio of 1:1 between the two regioisomers.

[0729] Step 15. Add NMP (1 mL) to (R)-3-methyl-4-(4-(1-(methanesulfonyl)cyclopropyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)morpholine (100 mg), 3-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole (145 mg), cesium carbonate (244 mg), and L-proline (21 mg) in a microwave-safe container, followed by CuBr (20 mg). Cap the container and degas (vacuum / argon, 3 cycles), then heat to 150 °C for 4 h. After cooling to rt, quench the reaction mixture with 20 mL of NH4Cl:H2O:NH4OH (4:3:1) and EtOAc (15 mL), filter through diatomaceous earth, and extract with ethyl acetate (2 × 150 mL). The combined organic extracts were washed with brine, dried over MgSO4, filtered, and concentrated to dryness. The residues were purified by silica gel chromatography (20-100% EtOAc / hexane) to give a mixture of regioisomers of (R)-3-methyl-4-(4-(1-(methanesulfonyl)cyclopropyl)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)morpholine (25 mg).

[0730] Step 16. Add TFA (0.2 mL) to a solution of (R)-3-methyl-4-(4-(1-(methanesulfonyl)cyclopropyl)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)morpholine (25 mg) in dichloromethane (1 mL). Stir the reaction mixture at rt for 18 h. Add toluene (10 mL) and remove volatiles under reduced pressure. Dissolve the residue in dioxane (3 mL) and saturated NaHCO3 aqueous solution (3 mL), heat the mixture to 65 °C for 18 h, then heat to 80 °C for 18 h. After cooling to rt, extract the mixture with dichloromethane (2 × 15 mL). Wash the combined organic extracts with brine, dry over MgSO4, filter, and concentrate to dryness. The residue was purified by silica gel chromatography (20-100% EtOAc / hexane) to obtain the desired product. The residue was suspended in CH3CN (1 mL) and water (1 mL) and lyophilized to give (R)-3-methyl-4-(4-(1-(methanesulfonyl)cyclopropyl)-1-(1H-pyrazol-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)morpholine (15 mg), which was a pale yellow foam. 1 H NMR (400MHz, CDCl3): δ7.66 (d; J=3.74Hz; 1H); 7.62 (d; J=2.36Hz; 1H); 6.98 (s ; 1H); 6.78 (s; 1H); 6.59 (d; J=3.76Hz; 1H); 4.32-4.37 (m; 1H); 4.05-4.09 (m; 1 H);3.87-3.91(m;1H);3.83-3.84(m;2H);3.64-3.71(m;1H);3.28-3.35(m;1H) ); 2.83 (s; 3H); 1.93-1.96 (m; 2H); 1.39-1.42 (m; 2H); 1.29 (d; J=6.71Hz; 3H). MS:[M+1]:402.2.

[0731] Intermediate A

[0732]

[0733] Step 1. Over 30 min, add hydrazine hydrate (8.7 mL) dropwise to a cold (0 °C) solution of acrylonitrile (12.4 mL) in THF (75 mL) to maintain an internal temperature below 10 °C. Stir the resulting mixture in an ice bath for 30 min, then warm to rt for 3 h. Cool the mixture again in an ice bath and add 2,4-dimethoxybenzaldehyde (31 g) over 10 min. Stir the resulting mixture in an ice bath for 25 min, warm to rt for 1 h, then concentrate under vacuum and place overnight under high vacuum with stirring to remove water.

[0734] The resulting residue was dissolved in n-BuOH (70 mL) and treated with NaOMe (20.4 g) to obtain a dark color and exothermic reaction. The mixture was heated to reflux for 1 h, cooled to room temperature, and poured into brine. EtOAc was added, and the organic layer was separated, washed with brine, dried over MgSO4, filtered through a diatomaceous earth mat, and concentrated under vacuum. The material was placed under high vacuum to remove residual n-BuOH. The process was repeated on the same scale, and the combined material was purified on silica gel (eluted with 1:1 EtOAc / hexane) to give 35 g of 1-(2,4-dimethoxybenzyl)-1H-pyrazole-5-amine.

[0735] Step 2. Add sodium diethyl oxaloacetate (16.1 g) to a solution of 1-(2,4-dimethoxybenzyl)-1H-pyrazol-5-amine (14 g) in AcOH (140 mL). Place the resulting suspension in an oil bath and heat to reflux for 2 h. Cool the reaction mixture in an ice bath and then slowly add it to 440 mL of cold water with rapid stirring. Stir the resulting suspension for 2 h, filter, wash with water and air dry overnight to give 19.2 g of ethyl 1-(2,4-dimethoxybenzyl)-6-hydroxy-1H-pyrazolo[3,4-b]pyridine-4-carboxylate as a yellow solid.

[0736] Step 3. At 0°C, pyridine (1.8 mL) was added to a suspension of 11.0 g of 1-(2,4-dimethoxybenzyl)-6-hydroxy-1H-pyrazolo[3,4-b]pyridine-4-carboxylate in acetonitrile (100 mL), followed by the addition of trifluorocarboxylic anhydride (3.8 mL) at a rate that maintained the internal temperature below 5°C. The reaction mixture was warmed to room temperature over 1 h, quenched with water (100 mL), and extracted with dichloromethane (2 × 100 mL). The combined organic extracts were washed with brine, dried over MgSO4, and concentrated under reduced pressure to dryness to give 10 g of 1-(2,4-dimethoxybenzyl)-6-(((trifluoromethyl)sulfonyl)oxy)-1H-pyrazolo[3,4-b]pyridine-4-carboxylate as a yellow solid, which was used in subsequent steps without further purification.

[0737] Step 4. At 0°C, (R)-3-methylmorpholine (6.8 g) and pyridine (2.0 mL) were added to a solution of crude 1-(2,4-dimethoxybenzyl)-6-(((trifluoromethyl)sulfonyl)oxy)-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (10 g) in DMF (100 mL). The reaction mixture was stirred at room temperature for 5 days, then diluted with water (100 mL) and EtOAc (120 mL). The layers were partitioned, and the aqueous layer was extracted with EtOAc (100 mL). The combined organic extracts were washed with brine, dried over MgSO4, filtered, and concentrated to dryness under vacuum. The residue was purified by ISCO CombiFlash (120 g column) (eluting with 10–100% EtOAc / hexane) to give 5.8 g of (R)-6-(3-methylmorpholino)-1H-pyrazolo[3,4-b]pyridine-4-carboxylic acid ethyl ester (intermediate A) as a yellow gel. LCMS (+ESI): m / z = 441.1 [M+H]+.

[0738] Compound 2

[0739] Step 1. At -78°C, MeMgBr (3M / Et2O, 1mL) was added to a solution of intermediate A (400mg) in THF (4mL), and the reaction mixture was warmed to room temperature. The reaction mixture was quenched with cooled saturated NH4Cl aqueous solution and extracted with EtOAc (2×30mL). The combined organic layers were washed with brine, dried over MgSO4, and concentrated to dryness under reduced pressure. The residue was purified by ISCO CombiFlash (40g column) (eluting with 10-100% EtOAc / hexane) to give 380mg of (R)-2-(1-(2,4-dimethoxybenzyl)-6-(3-methylmorpholino)-1H-pyrazolo[3,4-b]pyridin-4-yl)prop-2-ol as a yellow oil.

[0740] Step 2. At room temperature, TFA (1.36 mL) was added to a solution of (R)-2-(1-(2,4-dimethoxybenzyl)-6-(3-methylmorpholino)-1H-pyrazolo[3,4-b]pyridin-4-yl)prop-2-ol (380 mg) in dichloromethane (4 mL), and the solution was stirred for 18 h. The volatiles were removed under reduced pressure, and the residue was suspended in EtOAc (30 mL) and washed with saturated NaHCO3 aqueous solution. The aqueous layer was extracted with EtOAc (20 mL), and the combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated to dryness under reduced pressure to give 160 mg (R)-2-(6-(3-methylmorpholino)-1H-pyrazolo[3,4-b]pyridin-4-yl)prop-2-ol.

[0741] Step 3. Pack the microwave tube with (R)-2-(6-(3-methylmorpholino)-1H-pyrazolo[3,4-b]pyridin-4-yl)prop-2-ol (160 mg), SEM-protected 3-iodopyrazole (376 mg), Cs2CO3 (475 mg), L-proline (13 mg), CuBr (13 mg), and NMP (3 mL). Then cap the container and degas it (vacuum / argon, 3 cycles), and heat it to 150°C for 18 h. After cooling to room temperature, the reaction mixture was diluted with EtOAc (50 mL) and then purified by ISCO CombiFlash (24 g column) (eluting with 10–100% EtOAc / hexane) to give 100 mg of a yellow gel-like substance (R)-2-(6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)prop-2-ol. 1H-NMR and LCMS revealed two regiomeric isomers of the N-protected pyrazole from SEM.

[0742] Step 4. At room temperature, add TFA (0.211 mL) to a solution of (R)-2-(6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)prop-2-ol (100 mg) in dichloromethane (1 mL) and stir the reaction mixture for 18 h. Add toluene (10 mL) and remove volatiles under reduced pressure. Place the flask under high vacuum to remove residual TFA. Dissolve the residue in dioxane (3 mL) and add 1N NaOH (1 mL). Heat the reaction mixture to reflux for 3 h, cool to room temperature, and then dilute with EtOAc (20 mL) and water (20 mL). Distribute the layers and extract the aqueous layer with EtOAc (10 mL). Wash the combined organic layers with brine, dry with MgSO4, and concentrate to dryness under reduced pressure. The residue was adsorbed onto silica gel for purification by ISCO CombiFlash (12 g column) (eluting with 40-100% EtOAc / hexane). The desired product fraction was fractionated and concentrated to dryness. The residue was diluted in CH3CN (1 mL) and water (1 mL) for lyophilization to give 22 mg of the desired product (R)-2-(6-(3-methylmorpholino)-1-(1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)prop-2-ol as a colorless foam. Purity by HPLC at 254 nm: 93.0%. 1H NMR (400MHz, CDCl3): δ8.12 (s; 1H); 7.63 (d; J=2.10Hz; 1H); 6.83 (s; 1H); 6.72 (s; 1H); 4.45-4.49 (m; 1H); 4.02-4.09 (m; 2H) ); 3.77-3.86 (m; 3H); 3.66 (td; J = 11.90; 3.16Hz; 1H); 3.39 (td; J = 12.73; 3.87Hz; 1H); 1.73 (s; 6H); 1.35 (d; J = 6.74Hz; 3H).

[0743] Compound 3

[0744] Step 1. Add MeOH (0.062 mL) to a solution of intermediate A (3.4 g) in THF (35 mL). Heat the reaction mixture to 65 °C, then add 2 M LiBH4 in THF (5.8 mL) dropwise over 1 h. Stir the reaction mixture at 65 °C for 4 h, then cool to room temperature. Add acetone (1 mL) and stir at room temperature for 30 min. Dilute the mixture with a 1:1 saturated NH4Cl aqueous solution / water (80 mL) and EtOAc (80 × 100 mL). Distribute the layers and extract the aqueous layer with EtOAc (40 mL). Wash the combined organic extracts with brine, dry with MgSO4, and concentrate to dryness under reduced pressure. The residue was purified by ISCO CombiFlash (80 g column) (eluting with 30-100% EtOAc / hexane) to give (R)-(1-(2,4-dimethoxybenzyl)-6-(3-methylmorpholino)-1H-pyrazolo[3,4-b]pyridin-4-yl)methanol and (R)-(1-(2,4-dimethoxybenzyl)-6-(3-methylmorpholino)-2H-pyrazolo[3,4-b]pyridin-4-yl)methanol (a separable mixture of positional isomers).

[0745] Step 2. At 0 °C, triethylamine (0.141 mL) was added to a solution of (R)-(1-(2,4-dimethoxybenzyl)-6-(3-methylmorpholino)-2H-pyrazolo[3,4-b]pyridin-4-yl)methanol (600 mg) in dichloromethane (7 mL), followed by methanesulfonyl chloride (0.254 mL). The reaction mixture was stirred for 90 min at room temperature and then diluted with dichloromethane (40 mL) and water (40 mL). The layers were partitioned, the aqueous layer was extracted with dichloromethane (30 mL), and the combined organic extracts were washed with brine, dried over MgSO4, filtered, and concentrated to dryness to give 700 mg of (R)-(1-(2,4-dimethoxybenzyl)-6-(3-methylmorpholino)-2H-pyrazolo[3,4-b]pyridin-4-yl)methylmethanesulfonate, which was used in subsequent steps without further purification.

[0746] Step 3. LiI (393 mg) was added to a solution of (R)-(1-(2,4-dimethoxybenzyl)-6-(3-methylmorpholino)-2H-pyrazolo[3,4-b]pyridin-4-yl)methylmethanesulfonate (700 mg) in dioxane (7 mL). The mixture was heated to 50 °C under argon for 2.5 h. After cooling to room temperature, the mixture was diluted with EtOAc (50 mL) and water (50 mL). The layers were partitioned, and the aqueous layer was extracted with EtOAc (30 mL). The combined organic extracts were washed with 2M sodium bisulfite (50 mL), water (50 mL), and brine (50 mL), then dried over MgSO4, filtered, and concentrated under reduced pressure to dryness to give 760 mg (R)-4-(2-(2,4-dimethoxybenzyl)-4-(iodomethyl)-2H-pyrazolo[3,4-b]pyridin-6-yl)-3-methylmorpholine, which was used in subsequent steps without further purification.

[0747] Step 4. Add sodium methanesulfonate (190 mg) to a solution of (R)-4-(2-(2,4-dimethoxybenzyl)-4-(iodomethyl)-2H-pyrazolo[3,4-b]pyridin-6-yl)-3-methylmorpholine (790 mg) in DMF (8 mL). Stir the reaction mixture at room temperature for 2 h, then dilute with EtOAc (40 mL) and water (40 mL). Distribute the layers and extract the aqueous layer with EtOAc (30 mL). Wash the combined organic extracts with an aqueous solution of sodium thiosulfate (50 mL), water (50 mL), and brine, then dry to dryness with MgSO4, filter, and concentrate under reduced pressure to dryness. The residue was adsorbed onto silica gel for purification by Isco CombiFlash (40g column) (eluting with 30-100% EtOAc / hexane) to give 640 mg of (R)-4-(2-(2,4-dimethoxybenzyl)-4-((methanesulfonyl)methyl)-2H-pyrazolo[3,4-b]pyridin-6-yl)-3-methylmorpholine as a colorless foam.

[0748] Step 5. To a solution of (R)-4-(1-(2,4-dimethoxybenzyl)-4-((methanesulfonyl)methyl)-2H-pyrazolo[3,4-b]pyridin-6-yl)-3-methylmorpholine (640 mg) in toluene (3 mL), add TBAB (45 mg) and 1,2-dibromoethane (0.156 mL), followed by 50% NaOH (2.9 mL). Heat the reaction mixture to 60 °C for 2 h. Add another 1,2-dibromoethane (0.5 mL) and heat the mixture again at 60 °C for 18 h. After cooling to rt, dilute the mixture with EtOAc (30 mL) and water (25 mL), partition the layers, and extract the aqueous layer with EtOAc (20 mL). Wash the combined organic extracts with brine, dry to MgSO4, and concentrate to dryness under reduced pressure. The residue was adsorbed onto silica gel for purification by ISCO CombiFlash (24 g column) (eluting with 30-100% EtOAc / hexane) to give 510 mg of (R)-4-(1-(2,4-dimethoxybenzyl)-4-(1-methylsulfonyl)cyclopropyl)-2H-pyrazolo[3,4-b]pyridin-6-yl)-3-methylmorpholine as a pale yellow foam.

[0749] Step 6. (R)-3-methyl-4-(4-(1-(methanesulfonyl)cyclopropyl)-1H-pyrazolo[3,4-b]pyridin-6-yl)morpholine: At 0 °C, TFA (1.6 mL) was added to a solution of (R)-4-(1-(2,4-dimethoxybenzyl)-4-(1-methanesulfonyl)cyclopropyl)-2H-pyrazolo[3,4-b]pyridin-6-yl)-3-methylmorpholine (510 mg) in dichloromethane (5 mL). The reaction mixture was warmed to room temperature and stirred for 5 h. Toluene (10 mL) was added to the reaction mixture, and the volatiles were removed under vacuum, followed by co-evaporation with toluene (10 mL). The residue was dissolved in EtOAc (50 mL) and a saturated aqueous solution of NaHCO3 (40 mL) under vigorous stirring. The layers were partitioned, and the aqueous layer was extracted with EtOAc (30 mL). The combined extracts were washed with brine, dried over MgSO4, filtered, and concentrated under vacuum to dryness to obtain 350 mg of (R)-3-methyl-4-(4-(1-(methanesulfonyl)cyclopropyl)-1H-pyrazolo[3,4-b]pyridin-6-yl)morpholine, which was used in the next step without further purification.

[0750] Step 7. Pack a microwave tube with (R)-3-methyl-4-(4-(1-(methanesulfonyl)cyclopropyl)-1H-pyrazolo[3,4-b]pyridin-6-yl)morpholine (160 mg), pyrazole (310 mg), Cs₂CO₃ (390 mg), L-proline (11 mg), CuBr (11 mg), and NMP (2 mL). Cap the container and degas (vacuum / argon, 3 cycles), then heat to 150 °C for 18 h. After cooling to room temperature, dilute the reaction mixture with EtOAc (20 mL) and NH₄Cl:H₂O:NH₄OH (4:3:1, 20 mL), then filter through diatomaceous earth. Separate the layers and extract the aqueous layer with EtOAc (20 mL). Wash the combined organic layers with brine, dry with MgSO₄, filter, and concentrate to dryness under vacuum. The residue was adsorbed onto silica gel for purification by ISCO CombiFlash (24 g column) (eluting with 20-100% EtOAc / hexane) to give (R)-3-methyl-4-(4-(1-(methanesulfonyl)cyclopropyl)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-5-yl)-1H-pyrazolo[3,4-b]pyridin-6-yl)morpholine and the corresponding SEM-pyrazol regioisomer.

[0751] Step 8. To a solution of (R)-3-methyl-4-(4-(1-(methanesulfonyl)cyclopropyl)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-5-yl)-1H-pyrazolo[3,4-b]pyridin-6-yl)morpholine (37 mg) in dichloromethane (1 mL), add TFA (0.319 mL) and stir the reaction mixture for 18 h. Add toluene (10 mL) and remove volatiles under reduced pressure. Dissolve the residue in dioxane (3 mL) and saturated NaHCO3 aqueous solution (3 mL), and heat the mixture to 65 °C for 4 h, then to 80 °C for 18 h. After cooling to room temperature, extract the mixture with dichloromethane (2 × 15 mL), wash the combined organic extracts with brine, dry to MgSO4, filter, and concentrate to dryness. The residue was purified by silica gel rapid chromatography (eluting with EtOAc and 5% MeOH / EtOAc). The resulting residue was suspended in CH3CN (2 mL) and water (2 mL) and lyophilized to give 23 mg of (R)-3-methyl-4-(4-(1-(methanesulfonyl)cyclopropyl)-1-(1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-6-yl)morpholine as a pale yellow foam. 1 H NMR (400MHz, CDCl3): δ8.11 (s; 1H); 7.70 (d; J = 2.26Hz; 1H); 6.93 (d; J = 2.26Hz; 1H); 6.84 (s; 1H); 4.41-4.43 (m; 1H); 4.06-4.09 (m; 2H) ; 3.77-3.87 (m; 2H); 3.61-3.68 (m; 1H); 3.33-3.40 (m; 1H); 2.85 (s; 3H); 1.97-2.00 (m; 2H); 1.41-1.44 (m; 2H); 1.35 (d; J=6.78Hz; 3H). [M+1]:m / z 403.1.

[0752] Compound 4

[0753] Step 1. At 0°C, TFA (20 mL) was added to a solution of intermediate A (5.8 g, 13.167 mmol) in dichloromethane (60 mL). The reaction mixture was warmed to room temperature and stirred for 18 h. Toluene (60 mL) was added, and the volatiles were removed under vacuum and co-evaporated with toluene (20 mL). The residue was dissolved in dichloromethane (300 mL) and then treated with a saturated aqueous solution of NaHCO3 (200 mL) under vigorous stirring. The layers were separated, and the aqueous layer was extracted with dichloromethane (150 mL). The combined extracts were washed with brine, dried over MgSO4, filtered, and concentrated to dryness under vacuum to give 3.8 g of (R)-6-(3-methylmorpholino)-1H-pyrazolo[3,4-b]pyridine-4-carboxylic acid ethyl ester as a yellow solid, which was used in the next step without further purification.

[0754] Step 2. A mixture of (R)-6-(3-methylmorpholino)-1H-pyrazolo[3,4-b]pyridine-4-carboxylic acid ethyl ester (3.8 g), pyrazole (6.37 g), Cs₂CO₃ (10.7 g), L-proline (300 mg), CuBr (292 mg), and NMP (40 mL) was degassed (vacuum / argon, 3 cycles) and heated to 150 °C for 18 h. After cooling to room temperature, the reaction mixture was diluted with 10% citric acid to adjust the pH to approximately 6-7, and EtOAc (350 mL) was added. The mixture was filtered through diatomaceous earth and washed with EtOAc. The layers were partitioned, and the aqueous layer was extracted with EtOAc (150 mL). The combined organic layers were washed with brine, dried over MgSO₄, filtered, and concentrated to dryness under vacuum. The residue was purified by silica gel rapid chromatography (eluting with 0-10% MeOH / dichloromethane) to obtain 3.6 g of (R)-6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxylic acid and its SEM regioisomers as a yellow oil.

[0755] Step 3. Potassium carbonate (2.7 g) was added to a solution of (R)-6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxylic acid (3.6 g) in DMF (36 mL), followed by iodomethane (0.6 mL). The reaction mixture was stirred at room temperature for 18 h. EtOAc (50 mL) and water (50 mL) were added to separate the layers, and the aqueous layer was extracted with EtOAc (40 mL). The combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated to dryness under vacuum. The residue was purified by ISCO CombiFlash (80 g column) (eluting with 0-70% EtOAc / hexane) to give 2.2 g of (R)-6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxylic acid methyl ester as a yellow solid.

[0756] Step 4. Add lithium borohydride (3.4 mL) to a solution of (R)-6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (2.2 g) in THF (20 mL) and MeOH (0.038 mL). Heat the mixture to 65 °C for 4 h, then cool to room temperature. Add acetone (1 mL) and stir for 30 min. Dilute the mixture with (1:1) NH4Cl / water (50 mL) and then extract with EtOAc (2 × 40 mL). The combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated under vacuum to dryness to obtain 2 g of (R)-(6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)methanol, which was used without further purification.

[0757] Step 5. At 0°C, Et3N (0.69 mL) was added to a solution of (R)-(6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)methanol (2 g) in dichloromethane (20 mL), followed by MsCl (0.38 mL). The reaction mixture was then stirred at room temperature for 2 h. The mixture was diluted with dichloromethane (60 mL) and water (60 mL). The layers were partitioned, and the aqueous layer was extracted with dichloromethane (30 mL). The combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated under vacuum to obtain 2.3 g of (R)-(6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)methylmethanesulfonate, which was used in the next step without further purification.

[0758] Step 6. At room temperature, add NaCN (325 mg) to a solution of (R)-(6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)methylmethanesulfonate (2.3 g) in DMF (18 mL). Stir the reaction mixture for 18 h, then dilute with EtOAc (40 mL) and water (40 mL). Distribute the layers and extract the aqueous layer with EtOAc (35 mL). Wash the combined organic layers with brine, dry over MgSO4, filter, and concentrate under vacuum. The residue was purified by ISCO CombiFlash (24 g column) (eluting with 20-100% EtOAc / hexane) to give 440 mg (R)-2-(6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)acetonitrile.

[0759] Step 7. At 0°C, 0.148 mL of iodomethane was added to a solution of (R)-2-(6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)acetonitrile (430 mg) in THF (5 mL), followed by dropwise addition of potassium tert-butoxide (2.37 mL) over 10 min. The reaction mixture was stirred at 0°C for 1 h, then poured into a saturated aqueous solution of NH4Cl and extracted with EtOAc (2 × 35 mL). The combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated to dryness under vacuum. The residue was adsorbed onto silica gel for purification by ISCO CombiFlash (24g Gold SiO2 column) (eluting with 10-90% EtOAc / hexane) to give 140 mg (R)-2-methyl-2-(6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)propionitrile.

[0760] Step 8. Add 4 mg of hydrogenated (dimethylphosphonic acid-kP)[hydrobis(dimethylphosphonic-kP)]platinum(II) to a solution of (R)-2-methyl-2-(6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)propionitrile (90 mg) in EtOH / H2O (2 mL / 0.4 mL). Heat the mixture to 80 °C, then cool and concentrate to dryness. The residue was adsorbed onto silica for purification by ISCO CombiFlash (12g Gold SiO2 column) (eluted with 30-100% EtOAc / hexane) to give 82 mg of (R)-2-methyl-2-(6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)propionamide as a colorless solid.

[0761] Step 9. To a solution of (R)-2-methyl-2-(6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)propionamide (81 mg) in dichloromethane (2 mL), add TFA (0.30 mL) and stir the reaction mixture at room temperature for 18 h. Add additional TFA (0.5 mL) and stir the mixture for 6 h. Add toluene (10 mL) and remove volatiles under reduced pressure. Dilute the residue in 5 mL of MeOH / water (85:15) and stir at room temperature for 18 h. Remove volatiles under reduced pressure and dissolve the residue in EtOAc (25 mL) and treat with a saturated aqueous solution of NaHCO3 (20 mL). Distribute the layers and extract the aqueous layer with EtOAc (20 mL). The combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated to dryness under vacuum. The residue was adsorbed onto silica gel for purification by ISCO CombiFlash (12g Gold SiO2 column) (eluting with 80-100% EtOAc / hexane) to give 23 mg of (R)-2-methyl-2-(6-(3-(methylmorpholino)-1-(1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)propionamide as a colorless foam. 1 H NMR (400MHz, DMSO): δ12.80 (s; 1H); 7.90 (s; 1H); 7.83 (s; 1H); 7.06 (s; 1H); 7 .02 (s; 1H); 6.76-6.77 (m; 1H); 6.65 (s; 1H); 4.47-4.50 (m; 1H); 4.06 (d; J=13. 56Hz; 1H); 3.99 (d; J = 11.46Hz; 1H); 3.78 (d; J = 11.34Hz; 1H); 3.63-3.66 (m; 1H ); 3.47-3.53 (m; 1H); 3.17-3.23 (m; 1H); 1.54 (s; 6H); 1.22 (d; J=6.68Hz; 3H). MS(+ESI):m / z370.2.

[0762] Compound 5

[0763] Step 1. Tributylphosphine (0.2 mL) and TMAD (133.6 mg) were added to a solution of (R)-(6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)methanol (115 mg) and 2-hydroxyisobutyronitrile (0.07 mL) in anhydrous toluene (10 mL), and the resulting mixture was stirred at room temperature for 1 h, then diluted with water and extracted with EtOAc. The organic extract was dried and concentrated to dryness, and then purified by Combi-Flash (12g column) (eluting with 10-80% EtOAc / hexane) to give 110 mg of (R)-2-(6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)acetonitrile as a pale yellow oil.

[0764] Step 2. To a solution of (R)-2-(6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)acetonitrile (68 mg) in toluene (2 mL), add tetrabutylammonium bromide (9.66 mg) and 50% NaOH (0.5 mL), followed by 1,5-dibromopentane (0.027 mL). Heat the mixture to 65 °C for 2 h, then dilute with water and extract with EtOAc. The combined organic extracts were dried over NaSO4, concentrated to dryness, and purified by Combi-Flash (4g column) (eluting with 20-80% EtOAc / hexane) to give 54 mg of a pale yellow oil (R)-1-(6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)cyclohexane-1-carboxylonite.

[0765] Step 3. Add TFA (0.27 mL) to a solution of (R)-1-(6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)cyclohexane-1-carboxynitrile (54 mg) in dichloromethane (2 mL). Stir the reaction mixture at room temperature for 18 h, then concentrate under reduced pressure. Dissolve the residue in 5 mL MeOH / H2O (85:15) and stir at room temperature for 18 h, then concentrate. Dissolve the residue in EtOAc (25 mL) and a saturated aqueous solution of NaHCO3 (25 mL). Distribute the layers and extract the aqueous layer with EtOAc (10 mL). Wash the combined organic layers with brine, dry over MgSO4, filter, and concentrate. The residue was adsorbed onto silica gel for purification by ISCO CombiFlash (4g column) (eluting with 30-100% EtOAc / n-hexane) to give 11 mg of (R)-1-(6-(3-methylmorpholino)-1-(1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)cyclohexane-1-carboxylonite as a grayish-white foam. 1 HNMR (400MHz, CDCl3): δ1.37 (d; 3H); 1.96 (d; 5H); 2.09 (t; 2H); 2.29 (d; 2H); 3.39 (td; 1H); 3.65 (td; 1 H); 3.87-3.77 (m; 2H); 4.08 (d; 3H); 4.49 (d; 1H); 6.82 (s; 1H); 6.99 (d; 1H); 7.80 (d; 1H); 8.22 (s; 1H).

[0766] Compound 6

[0767] Step 1. At 0 °C, mCPBA (51.41 g) was added in portions over 30 min to a mechanically stirred suspension of 4-chloro-1H-pyrrolo[2,3-b]pyridine (35 g) in EtOAc (600 mL). The reaction mixture was then stirred at rt for 18 h, and the solid was collected by filtration and washed with n-heptane (350 mL). The residue was dried under high vacuum to give 62 g of 4-chloro-1H-pyrrolo[2,3-b]pyridine 7-oxide 3-chlorobenzoate as a gray solid.

[0768] Step 2. Dimethyl sulfate (9.6 mL) was added to a mixture of 30 g of 4-chloro-1H-pyrrolo[2,3-b]pyridine 7-oxide 3-chlorobenzoate in acetonitrile (300 mL), and the reaction mixture was heated to 60 °C for 18 h. After cooling to rt, (R)-3-methylphosphine (14 g) was added, followed by diisopropylethylamine (48.2 mL), and the reaction mixture was heated to 60 °C for 18 h. After cooling to rt, the volatiles were removed under vacuum, and the residue was purified by silica gel column chromatography (eluting with 10-40% EtOAc / hexane) to give 12 g of (R)-4-(4-chloro-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-methylmorpholine as a pale gray solid.

[0769] Step 3. A mixture of (R)-4-(4-chloro-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-methylmorpholine (11.64 g), iodopyrazole (15.11 g), CuI (81 mg), trans-N,N-dimethylcyclohexane-1,2-diamine (0.66 mL), and K3PO4 (17.23 g) in dioxane (110 mL) was purged three times with argon and heated to 110 °C for 18 h. The mixture was cooled and filtered through a silica gel pad (eluted with EtOAc (700 mL)). The filtrate was concentrated to dryness under vacuum and then purified by rapid silica gel chromatography (eluted with 10–25% EtOAc / hexane). The pure fraction was fractionated and concentrated to give 19.3 g.

[0770] A mixture of SEM regioisomers of (R)-4-(4-chloro-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-methylmorpholine.

[0771] Step 4. Add potassium acetate (3.32 g) to a solution of (R)-4-(4-chloro-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-methylmorpholine (5.0 g), bis(pinacol)diboron (4.25 g), Pd2(dba)3 (510 mg), and tricyclohexylphosphine (780 mg) in dioxane (70 mL). Purge the mixture with argon and heat to 100 °C overnight, then cool, dilute with ethyl acetate, and filter through a diatomaceous earth mat. Concentrate the filtrate to dryness and return it to the reaction conditions. After overnight, dilute the reaction mixture with ethyl acetate, filter through a diatomaceous earth mat, and concentrate to dryness. The product was purified by column chromatography (eluting with 0-50% ethyl acetate / hexane) to obtain 4.38 g of (R)-3-methyl-4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)morpholine as a yellow powder.

[0772] Step 5. To a blue flask containing (R)-3-methyl-4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)morpholine (106 mg), 2-bromophenylmethyl sulfone (93 mg), and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (10 mg), dioxane (1 mL) and 2N Na₂CO₃ (250 μL) were added. The mixture was evacuated, purged with argon (3x), and heated at 120 °C for 24 h. The mixture was then cooled and partitioned between water and ethyl acetate. The organic phase was separated and the aqueous phase was extracted three times with ethyl acetate. The combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated. The crude material was purified on a Redisep Gold Column (12 g) using 0-100% ethyl acetate / hexane to give 76 mg of (R)-3-methyl-4-(4-(2-(methylsulfonyl)phenyl)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)morpholine.

[0773] Step 6. Add TFA (0.45 mL) to a solution of (R)-3-methyl-4-(4-(2-(methanesulfonyl)phenyl)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)morpholine (76 mg) in dichloromethane (2 mL). Stir the reaction mixture overnight at room temperature, then concentrate and redissolve in 85 / 15 MeOH / H2O and stir again for 4 h. Concentrate the reaction mixture and partition between ethyl acetate and water. Separate the organic layers and extract the aqueous layer with ethyl acetate (3x). Wash the combined organic layers with brine, dry over Na2SO4, filter and concentrate. The crude material was purified on a Redisep column (24 g) (eluted with 40-60% ethyl acetate / hexane) to give 58 mg (R)-3-methyl-4-(4-(2-(methylsulfonyl)phenyl)-1-(1H-pyrazol-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)morpholine. 1 HNMR(d6-DMSO)δ12.7(s,1H),8.2(d 1H),7.8(m,1H),7.7(m,2H),7.6(m1H),7.5(m 1H),7.0(s,1H),6.7(s,1H),4.3(m,1H),4.0(m,1H),3.7(m,2H),3.5(m,1H),3.2(m,1H),2.9(s,3H),1.2(d 3H).

[0774] Compound 7

[0775] Step 1. To a solution of 508 mg of 4-chloro-3-methyl-1H-pyrrolo[2,3-b]pyridine 7-oxide 3-chlorobenzoate in acetonitrile (10 mL), 275 mg of 3-chlorobenzoic acid and 0.29 mL of dimethyl sulfate were added, and the reaction mixture was heated at 60 °C for 36 h. After cooling, (R)-3-methylmorpholine (423 mg) and DIPEA (1.45 mL) were added, and the reaction mixture was heated at 60 °C for 26 h. The reaction mixture was concentrated and purified by silica gel chromatography using 40 to 100% ethyl acetate / hexane to give 243 mg of (R)-4-(4-chloro-3-methyl-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-methylmorpholine.

[0776] Step 2. Add a solution of 3-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole (2.79 g) in dioxane (20 mL) to a 100 mL flask containing (R)-4-(4-chloro-3-methyl-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-methylmorpholine (2.08 g). Puff argon gas into the mixture and add ground K3PO4 (2.9 g), followed by trans-N,N'-dimethylcyclohexane-1,2-diamine (111 mg) and CuI (15 mg). Heat the reaction mixture at 100 °C for 44 h, then filter through diatomaceous earth and wash with ethyl acetate. Wash the filtrate with water, and dry the organic layer with Na2SO4, filter, and concentrate. Purification on a Redisep Gold column (80 g) (eluted with 0-100% ethyl acetate / hexane) yielded 2.53 g of a mixture of regioisomers of (R)-3-methyl-4-(6-(1-(methanesulfonyl)cyclopropyl)-2-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)thio)pyrimidin-4-yl)morpholine.

[0777] Step 3. To a solution of (R)-4-(4-chloro-3-methyl-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-methylmorpholine (138 mg) in THF (1 mL), add isobutyronitrile (350 μL), followed by LiHMDS (1 M in THF, 2.7 mL). Microwave the mixture at 100 °C for 15 min, then cool and partition between a saturated NH4Cl aqueous solution and ethyl acetate. Extract the aqueous layer 3x with ethyl acetate and wash the combined organic layers with brine, dry over Na2SO4, filter, and concentrate. Purification on a Redisep Gold column (24 g) using 0-100% ethyl acetate / hexane yielded 136 mg of (R)-2-methyl-2-(3-methyl-6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrrolo[2,3-b]pyridin-4-yl)propionitrile in an oily form.

[0778] Step 4. Add TFA (250 μL) to a solution of (R)-2-methyl-2-(3-methyl-6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrrolo[2,3-b]pyridin-4-yl)propionitrile (135 mg) in dichloromethane (1 mL). Stir the reaction mixture at room temperature for 3 days, then concentrate and partition between ethyl acetate and a saturated aqueous solution of NaHCO3. Separate the organic layers and extract the aqueous layer with ethyl acetate (3x). Dry the combined organic layers with Na2SO4, filter, and concentrate. Purification on a Redisep Gold column (12 g) using 30-100% ethyl acetate / hexane yielded 15 mg of (R)-2-methyl-2-(3-methyl-6-(3-methylmorpholino)-1-(1H-pyrazol-3-yl)-1H-pyrrolo[2,3-b]pyridin-4-yl)propionitrile. 1 H NMR(d6-DMSO)δ12.7(s,1H),7.8(s,1H),7.6(s,1H),6.9(s,1H),6.6(s,1H),5.7(s,1H),4.4(m ,1H),4.0(m,1H),3.9(m,1H),3.8(m,1H),3.7(m,1H),3.5(m,1H),3.2(m,1H),2.6(s,3H),1.2(d 3H).

[0779] Compound 8

[0780] Step 1. Diisopropylethylamine (509 μL) was added to a solution of 5,7-dichloro-3H-imidazo[4,5-b]pyridine (457 mg) and 2-(chloromethoxy)ethyltrimethylsilane (516 μL) in DMF (8 mL), and the mixture was stirred at room temperature for 1 h. Water and Et2O were added, and the phases were separated. The aqueous phase was extracted with Et2O (2x), and the combined organic extracts were washed with brine, dried over Na2SO4, filtered, and evaporated under reduced pressure. The crude mixture was purified by silica gel chromatography (eluting with 0 to 70% EtOAc / hexane) to give 473 mg of 2-[((5,7-dichloroimidazolo[4,5-b]pyridin-3-yl)methoxy]ethyltrimethylsilane (provisional partition) and 120 mg of 2-[(5,7-dichloroimidazolo[4,5-b]pyridin-1-yl)methoxy]ethyltrimethylsilane (provisional partition). Major isomers: 1¹H NMR (400MHz, CDCl₃) δ 8.22 (s, ¹H), 7.35 (s, ¹H), 5.64 (s, 2H), 3.69–3.48 (m, 2H), 0.99–0.85 (m, 2H), -0.04 (s, 9H). LCMS: 318.12 (M+H). Minor isomers: 1 HNMR (400MHz, CDCl3) δ8.21(s,1H),7.30(s,1H),5.74(s,2H),3.68–3.42(m,2H),1.05–0.84(m,2H),-0.07(s,9H). LCMS:319.97(M+H). LCMS:318.25(M+H).

[0781] Step 2. Under nitrogen atmosphere, add Pd(dppf)Cl2·CH2Cl2 (31 mg) to a solution of 2-[(5,7-dichloroimidazolo[4,5-b]pyridin-3-yl)methoxy]ethyl-trimethylsilane (90 mg), K3PO4 (2 M, 424 μL), and (2-methylsulfonylphenyl)boronic acid (68 mg) in dioxane (1 mL), and stir overnight at 80 °C. Add water and EtOAc, and separate the phases. Extract the aqueous phase with EtOAc (2x), and wash the combined organic extracts with brine, dry with Na2SO4, filter, and evaporate under reduced pressure. The crude mixture was purified by silica gel chromatography (eluting with 0 to 100% EtOAc / hexane) to give a 1:1 mixture of 2-[[7-chloro-5-(2-methylsulfonylphenyl)imidazo[4,5-b]pyridyl-3-yl]methoxy]ethyl-trimethylsilane as regioisomers. 1 H NMR (400MHz, CDCl3) δ8.22(dd,J=7.7,1.6Hz,1H),8.14(s,1H),7.79–7.64(m,2H),7.41(dd,J=7.3,1. 6Hz,1H),7.33(s,1H),5.67(s,2H),3.77–3.63(m,2H),3.03(s,3H),1.04–0.91(m,2H),-0.03(s,9H). LCMS:437.94(M+H).

[0782] Step 3. To a solution of 2-[[7-chloro-5-(2-methylsulfonylphenyl)imidazo[4,5-b]pyridyl-3-yl]methoxy]ethyl-trimethylsilane (640 mg) in anhydrous dioxane (1 mL), cesium carbonate (952 mg), RuPhos PdG1 methyl tert-butyl ether adduct (119 mg), and (3R)-3-methylmorpholine (332 μL) were added. The mixture was purged with nitrogen and then heated to 100 °C in a sealed vial for 16 h. Water and EtOAc were added, and the phases were separated. The aqueous phase was extracted with EtOAc (2x), and the combined organic extracts were washed with brine, dried over Na2SO4, filtered, and evaporated under reduced pressure. The residue was absorbed in DMSO and purified by reverse chromatography to obtain 490 mg of trimethyl-[2-[[5-[(3R)-3-methylmorpholin-4-yl]-7-(2-methylsulfonylphenyl)imidazo[4,5-b]pyridin-3-yl]methoxy]ethyl]silane. 1 H NMR (400MHz, CDCl3) δ8.21 (dd, J=7.9, 1.4Hz, 1H), 7.95 (s, 1H), 7.68 (td, J=7.5, 1.4Hz, 1H), 7.59 (td, J=7 .7,1.5Hz,1H),7.52(dd,J=7.5,1.4Hz,1H),6.62(s,1H),5.56(d,J=2.3Hz,2H),5.06(s,1H),4.33(d,J=13 .2Hz,1H),4.03(dd,J=11.4,3.6Hz,1H),3.93(dd,J=11.4,3.1Hz,1H),3.84–3.71(m,2H),3.63–3.53(m,2 H),3.49(td,J=6.5,5.5,3.8Hz,1H),3.32(s,3H),1.34(d,J=6.7Hz,3H),0.97–0.83(m,2H),-0.06(s,9H). LCMS:505.19(M+H).

[0783] Step 4. Slowly add 250 μL of TFA to a solution of trimethyl-[2-[[5-[(3R)-3-methylmorpholin-4-yl]-7-(2-methylsulfonylphenyl)imidazo[4,5-b]pyridin-3-yl]methoxy]ethyl]silane (55 mg) in dichloromethane (1 mL), and stir the mixture overnight at room temperature. Add another 250 μL of TFA and stir the mixture over the weekend. Remove volatiles under reduced pressure, dissolve the crude residue in EtOAc and treat with saturated NaHCO3 solution, then separate the layers. Extract the aqueous layer with EtOAc (2x), and dry the combined organic extracts with sodium sulfate, filter and concentrate. The residue was purified by reversed-phase chromatography to obtain 31 mg of (3R)-3-methyl-4-[7-(2-methylsulfonylphenyl)-3H-imidazo[4,5-b]pyridin-5-yl]morpholine. 1 H NMR(400MHz, CDCl3)δ8.23(dd,J=7.9,1.4Hz,1H),7.74(s,1H),7.70(td,J=7.5,1.4Hz, 1H),7.63(td,J=7.7,1.5Hz,1H),7.48(dd,J=7.5,1.4Hz,1H),6.73(s,1H),4.25(q,J=7. 0Hz,1H),4.02(dd,J=11.4,3.6Hz,1H),3.92–3.84(m,1H),3.80(d,J=2.1Hz,2H),3.64(t d,J=11.7,3.0Hz,1H),3.27(td,J=12.5,3.8Hz,1H),2.97(s,3H),1.27(d,J=6.7Hz,3H). LCMS:374.08(M+H).

[0784] Step 5. Under nitrogen atmosphere, copper bromide (45 mg) was added to a solution of (3R)-3-methyl-4-[5-(2-methylsulfonylphenyl)-3H-imidazo[4,5-b]pyridin-7-yl]morpholine (290 mg), 2-[(3-iodopyrazol-1-yl)methoxy]ethyl-trimethyl-silane (510 mg), 3-(1,1-difluoroethyl)benzenesulfinic acid (54 mg), and cesium carbonate (634 mg) in NMP (3.5 mL), and the mixture was heated overnight at 120 °C. The mixture was cooled, treated with a saturated aqueous solution of NH4Cl, water, and ammonium hydroxide (4:1:3), and extracted with EtOAc. The aqueous phase was extracted with EtOAc (2x), and the combined organic phases were washed with brine, dried over Na2SO4, filtered, and concentrated. The crude product was purified by reversed-phase chromatography to give 220 mg of a mixture of regioisomers of trimethyl-[2-[[3-[5-[(3R)-3-methylmorpholin-4-yl]-7-(2-methylsulfonylphenyl)imidazo[4,5-b]pyridin-3-yl]pyrazol-1-yl]methoxy]ethyl]silane. LCMS: 569.38 (M+H).

[0785] Step 6. Add TFA (56 μL) to a solution of trimethyl-[2-[[3-[5-[(3R)-3-methylmorpholin-4-yl]-7-(2-methylsulfonylphenyl)imidazo[4,5-b]pyridin-3-yl]pyrazol-1-yl]methoxy]ethyl]silane (14 mg) in dichloromethane (1 mL) and stir the mixture overnight at room temperature. Remove volatiles under reduced pressure, dissolve the mixture in dioxane (1 mL), and alkalize to pH approximately 10 using 3N NaOH, heating at 80 °C for 3 h. Partition the mixture between EtOAc and water. Extract the aqueous phase with EtOAc (2x), and wash the combined organic extracts with brine, dry over Na2SO4, filter, and concentrate. The residue was absorbed in DMSO and purified by reverse chromatography to give 3.7 mg of (3R)-3-methyl-4-[7-(2-methylsulfonylphenyl)-3-(1H-pyrazol-3-yl)imidazo[4,5-b]pyridin-5-yl]morpholine. 1H NMR (400MHz, DMSO-d6) δ13.03(s,1H),8.47(s,1H),8.14(dd,J=7.8,1.5Hz,1H),7.96(d ,J=2.3Hz,1H),7.80(dtd,J=21.7,7.5,1.5Hz,2H),7.52(dd,J=7.4,1.5Hz,1H),6.97(d ,J=2.2Hz,1H),6.79(s,1H),4.42–4.32(m,1H),3.99(d,J=11.8Hz,2H),3.82–3.65(m,2 H), 3.54 (td, J = 11.7, 3.0 Hz, 1H), 3.19 (s, 3H), 3.18–3.07 (m, 1H), 1.19 (d, J = 6.6 Hz, 3H). LCMS:438.94(M+H).

[0786] Intermediate C

[0787] Step 1. To a solution of 24.7 g (297 mmol) of 3-aminopyrazole in 181 mL of 6 N HCl at -5 °C, add 300 mL (297 mmol) of NaNO2 aqueous solution. Then, add dropwise a solution of 113 g (595 mmol) of SnCl2 in 510 mL of concentrated HCl, and stir the resulting mixture at room temperature for 2 h. Evaporate the solvent under reduced pressure to obtain a light brown solid of 3-hydrazino-3H-pyrazole, which was used as is without further purification. 1 H NMR (400MHz, DMSO-d6, δppm): 9.90 (s, 3H), 7.65 (d, J = 2.4Hz, 1H), 5.81 (d, J = 2.3Hz, 1H).

[0788] Step 2. Load 500 mL of frame-dried RBF with 2,6-difluoro-4-iodopyridine (17 g, 70.5 mmol) and anhydrous THF (255 mL). Cool the yellow reaction mixture to -78 °C and add commercially available LDA (1.0 M in THF / hexane, 84.7 mL, 84.7 mmol) dropwise at a rate that keeps the internal temperature below -68 °C. Stir the pale brown solution at -78 °C for 1 h, then add ethyl formate (8.5 mL, 105.678 mmol) over 10 min. Monitor the reaction by TLC and complete after 30 min. Add formic acid (5.3 mL, 140.5 mmol) dropwise and stir the mixture at -78 °C for 10 min, then dilute with EtOAc (150 mL). Warm the mixture to 0 °C and add water (100 mL). Separate the layers and extract the aqueous layer with EtOAc (150 mL). The combined organic layers were washed with brine, dried with Na2SO4, filtered, and concentrated under vacuum to obtain 19g of 2,6-difluoro-4-iodo-pyridine-3-carboxaldehyde, which was a light brown solid. 1 H NMR: (400MHz, CDCl3), δ10.11 (s, 1H), 7.54 (d; J=2.87Hz; 1H).

[0789] Step 3. Add 2,6-difluoro-4-iodopyridine-3-carboxaldehyde (4.4 g, 16.3 mmol) to a suspension of 3-hydrazine-3H-pyrazole (12.5 g, 94.3 mmol) in 95% EtOH (70 mL) and stir the mixture at rt for 15 min. Then remove most of the volatiles under reduced pressure. Dissolve the orange mixture in EtOAc and NaHCO3 and stir at rt for 15 min, resulting in vigorous gas escape. Separate the phases and extract the aqueous phase three times with EtOAc. Wash the combined organic extracts with water and brine, dry over MgSO4, filter, and evaporate under reduced pressure to give (E)-3-((2-(1H-pyrazol-3-yl)hydrazine)methyl)-2,6-difluoro-4-iodopyridine (5.5 g, 15.9 mmol) as a yellow / orange solid. 1 H NMR (400MHz, DMSO-d6)12.02(s,1H),10.89(s,1H),7.92(s,1H),7.82(d,1H),7.54(s,1H),5.97(s,1H).

[0790] Step 4. The solution of (E)-3-((2-(1H-pyrazol-3-yl)hydrazine)methyl)-2,6-difluoro-4-iodopyridine (8.6 g, 24.7 mmol) in NMP (115 mL) was divided into 20 mL batches and heated in a microwave reactor at 200 °C for 20 min. The combined mixture was then added dropwise to water with vigorous stirring to obtain a turbid mixture, which was stirred at rt for 5 min and then cooled to 0 °C. The precipitate was filtered, washed with water, and dried over a Buchner funnel for 1 h and under reduced pressure for 1 h to obtain 6-fluoro-4-iodo-1-(1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridine (6.8 g, 20.7 mmol) as a light brown powder. 1 H NMR (400MHz, DMSO-d6) δ13.13(s,1H),8.29(s,1H),7.95(t,1H),7.71(d,1H),6.67(t,1H).

[0791] Step 5. A solution of 6-fluoro-4-iodo-1-(1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridine (6.8 g, 20.7 mmol) and (R)-3-methylmorpholine (1.24 mL, 7.23 mmol) in DMSO (35 mL) was sealed in a thick-walled tube and heated to 120 °C for 45 min. The mixture was then added dropwise to a water-filled conical flask with vigorous stirring. The turbid mixture was stirred at rt for 5 min and then at 0 °C for 20 min. The precipitate was filtered through a Buchner funnel, washed with water, and dried overnight on a Buchner funnel to give (R)-4-(4-iodo-1-(1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-6-yl)-3-methylmorpholine (6.9 g, 16.8 mmol), intermediate B.

[0792] Step 6. Add 2-(chloromethoxy)ethyltrimethylsilane (1.04 mL, 5.8 mmol) to a solution of intermediate B (2.00 g, 4.88 mmol) in DMF (20 mL), followed by diisopropylethylamine (1.28 mL, 7.3 mmol), and stir the resulting mixture for 40 min. Partition the mixture between EtOAc and water, and extract the aqueous phase with EtOAc (2x). Wash the combined organic layers with water (2x) and brine, then dry over Na2SO4, filter, and evaporate. Purified by silica gel chromatography (gradient 0 to 80% EtOAc / hexane), 2-[[3-[4-iodo-6-[(3R)-3-methylmorpholin-4-yl]pyrazolo[3,4-b]pyridin-1-yl]pyrazol-1-yl]methoxy]ethyltrimethylsilane (0.67 g, 1.25 mmol) and 2-[[5-[4-iodo-6-[(3R)-3-methylmorpholin-4-yl]pyrazolo[3,4-b]pyridin-1-yl]pyrazol-1-yl]methoxy]ethyltrimethylsilane (0.17 g, 0.31 mmol) were obtained.

[0793] Compound 86

[0794] Step 1. 2,6-Difluoro-4-iodo-pyridin-3-carboxaldehyde (1.76 g, 6.54 mmol) in DME (15 mL) was dissolved in a round-bottom flask, and hydrazine hydrate (535 μL, 65% purity, 7.1 mmol) was added. The reaction mixture was stirred at rt for 4 h. Water was added to the heterogeneous yellow solution, and it was stirred at rt for 30 min. The resulting solid was then collected by filtration, washed with water, and dried under vacuum overnight to give 6-fluoro-4-iodo-1H-pyrazolo[3,4-b]pyridine.

[0795] Step 2. 6-Fluoro-4-iodo-1H-pyrazolo[3,4-b]pyridine (2.18 g, 8.29 mmol) was dissolved in 30 mL of DMSO in RBF. (3R)-3-methylmorpholine (3.43 g, 33.94 mmol, 3.85 mL) was added to this solution, and the reaction mixture was stirred overnight at 120 °C. The reaction mixture was then slowly cooled to rt. Water was slowly added over 5–10 minutes, and the flask was placed in an ice bath while the solution was stirred for 1 h. The resulting solid was then collected by filtration, washed with water, and dried under vacuum for 1 h, followed by overnight drying under vacuum to give (3R)-4-(4-iodo-1H-pyrazolo[3,4-b]pyridin-6-yl)-3-methylmorpholine (2.12 g, 6.16 mmol).

[0796] Step 3. 6-Fluoro-4-iodo-1H-pyrazolo[3,4-b]pyridine (1.54 g, 5.86 mmol) in DMF (40 mL) was dissolved in RBF and cooled to 0 °C. 60 wt% sodium hydride (281.0 mg, 7.03 mmol, 60% purity) was added to this solution, and the reaction mixture was stirred at 0 °C for 30 min. SEM-Cl (1.46 g, 8.78 mmol, 1.55 mL) was then added, and the solution was stirred at 0 °C for 5 min, then returned to room temperature and stirred for another 1 h. Saturated NH4Cl was added, followed by water, and the mixture was stirred for 30 min. The resulting solid was collected by filtration and dried under vacuum overnight to give a mixture of SEM regioisomers of 2-[(6-fluoro-4-iodo-pyrazolo[3,4-b]pyridin-2-yl)methoxy]ethyl-trimethyl-silane.

[0797] Step 4. 2-[[4-iodo-6-[(3R)-3-methylmorpholin-4-yl]pyrazolo[3,4-b]pyridin-1-yl]methoxy]ethyl-trimethylsilane (103 mg, 217.11 μmol) in THF (1 mL) was dissolved in RBF. 2-methylpropionitrile (150.15 mg, 2.17 mmol, 195 μL) was added to this solution, followed by LiHMDS (1 M, 1.09 mL). The reaction mixture was stirred at 20 °C for 15 min under microwave irradiation, then heated to 100 °C for 12 min. Water and EtOAc were added, and the phases were separated. The aqueous phase was extracted a second time with EtOAc. The combined organic phases were washed with a saturated brine solution, dried over MgSO4, filtered, and evaporated under reduced pressure. The crude product was purified using a 15.5 g Gold C18 Isco column and elution buffer of 10 to 100% water / MeCN to give 2-methyl-2-[6-[(3R)-3-methylmorpholin-4-yl]-1-(2-trimethylsilylethoxymethyl)pyrazolo[3,4-b]pyridin-4-yl]propionitrile.

[0798] Step 5. 2-Methyl-2-[6-[(3[R)-3-methylmorpholin-4-yl]-1-(2-trimethylsilylethoxymethyl)pyrazolo[3,4-b]pyridin-4-yl]propionitrile (690 mg, 1.66 mmol) in DCM (30 mL) was dissolved in RBF and TFA (3.80 mL, 50 mmol) was added. The reaction mixture was stirred overnight at rt, and then the volatiles were removed under vacuum. The crude product was dissolved in 1 mL DMSO and purified using a 15.5 g Gold C18 Isco column and elution with 5 to 100% water / MeCN to give 2-methyl-2-[6-[(3R)-3-methylmorpholin-4-yl]-1H-pyrazolo[3,4-b]pyridin-4-yl]propionitrile.

[0799] Step 6. A solution of 2-methyl-2-[6-[(3[R)-3-methylmorpholin-4-yl]-1H-pyrazolo[3,4-b]pyridin-4-yl]propionitrile (100 mg, 0.35 mmol), 5-iodo-3-methyl-1-tetrahydropyran-2-yl-pyrazole (205 mg, 0.7 mmol), cesium carbonate (285 mg, 0.87 mmol), and (1S,2S)-N1,N2-dimethylcyclohexane-1,2-diamine (100 mg, 0.70 mmol) in NMP (1.2 mL) was purged with nitrogen for 5 minutes, and then copper iodide (67 mg, 0.35 mmol) was added. The mixture was heated to 120 °C for 16 h. Water was added, the mixture was stirred for 30 minutes, and the resulting solid was collected by filtration and dried under vacuum for 1 h. The solid was then dissolved in 1 mL of DMSO and purified by reversed-phase chromatography (5 to 100% water / MeCN in 20 CV) to give 2-methyl-2-[6-[(3[R)-3-methylmorpholin-4-yl]-1-(5-methyl-2-tetrahydropyran-2-yl-pyrazol-3-yl)pyrazolo[3,4-b]pyridin-4-yl]propionitrile.

[0800] Step 7. In a round-bottom flask, 42 ​​mg, 93 μmol of 2-methyl-2-[6-[(3[R)-3-methylmorpholin-4-yl]-1-(5-methyl-2-tetrahydropyran-2-yl-pyrazol-3-yl)pyrazolo[3,4-b]pyridin-4-yl]propionitrile was dissolved in 0.5 mL of MeOH. 112 μL of HCl in MeOH was added to this solution, and the reaction mixture was stirred at 60 °C for 1 h. The evaporation was evaporated under reduced pressure, and the crude product was purified using a 15.5 g Gold C18 Isco column and 5–100% water / MeCN eluent to give compound 86. 1 H NMR (400MHz, DMSO-d6) δ12.51(s,1H),8.27(s,1H),6.73(s,1H),6.49(d,J=2. 0Hz,1H),4.46(s,1H),4.09–4.00(m,1H),3.97(dd,J=11.4,3.5Hz,1H),3.76( d,J=11.4Hz,1H),3.64(dd,J=11.5,3.1Hz,1H),3.49(td,J=11.9,3.1Hz,1H), 3.19(td,J=12.6,3.8Hz,1H),2.30(s,3H),1.85(s,6H),1.20(d,J=6.7Hz,3H).

[0801] Compound 99

[0802] Step 1. A solution of 2-[[5-[4-iodo-6-[(3[R)-3-methylmorpholin-4-yl]pyrazolo[3,4-b]pyridin-1-yl]pyrazol-1-yl]methoxy]ethyl-trimethylsilane (500 mg, 0.92 mmol) in THF (8 mL) was cooled to -78 °C and slowly treated with nBuLi (2.5 M, 0.48 mL). The mixture was stirred for 40 min. Then a solution of tetrahydropyran-3-one (27 μL, 2.78 mmol) in 1.5 mL THF was added to the mixture. The flask was removed from the dry ice bath and stirring was continued for 1 h. The mixture was then quenched with saturated NH4Cl solution and EtOAc was added, and the phases were separated. The aqueous phase was extracted twice more with EtOAc, and the combined organic extracts were washed with a saturated salt solution, dried over Na2SO4, filtered, and evaporated under reduced pressure to give 3-[6-[(3R)-3-methylmorpholin-4-yl]-1-[2-(2-trimethylsilylethoxymethyl)pyrazol-3-yl]pyrazolo[3,4-b]pyridin-4-yl]tetrahydropyran-3-ol.

[0803] Step 2. A solution of 3-[6-[(3[R)-3-methylmorpholin-4-yl]-1-[2-(2-trimethylsilylethoxymethyl)pyrazol-3-yl]pyrazolo[3,4-b]pyridin-4-yl]tetrahydropyran-3-ol (168 mg, 0.325 mmol), triethylsilane (291 mg, 2.50 mmol, 0.4 mL), DCM (1 mL), and TFA (5.96 g, 52.3 mmol, 4 mL) was stirred at rt for 10 min. Volatiles were removed under reduced pressure, and the residues were purified by reversed-phase chromatography to give a mixture of compounds 99 and 100, which were separated by SFC.

[0804] Compound 121

[0805] Step 1. The solution of intermediate C (200 mg, 0.37 mmol) in THF (4 mL) was cooled to -78 °C and slowly treated with nBuLi (2.5 M, 0.19 mL). The mixture was stirred for 40 min, and then a solution of 8-oxabicyclo[3.2.1]oct-3-one (27 μL, 1.2 mmol) in 0.4 mL THF was added. The flask was then removed from the dry ice bath and warmed to rt over 1.5 h. The mixture was then quenched with saturated NH4Cl solution and extracted with EtOAc. The aqueous phase was extracted twice more with EtOAc, and the combined organic extracts were washed with saturated brine, dried over Na2SO4, filtered, and evaporated under reduced pressure. The residue was used in the next step without further purification.

[0806] Step 2. A solution of 3-[6-[(3R)-3-methylmorpholin-4-yl]-1-[2-(2-trimethylsilylethoxymethyl)pyrazol-3-yl]pyrazolo[3,4-b]pyridin-4-yl]-8-oxabicyclo[3.2.1]oct-3-ol (100 mg, 0.18 mmol) and triethylsilane (0.23 mL, 1.42 mmol) in DCM (1 mL) was treated with TFA (2.3 mL, 30 mmol) under rt and stirred for 10 min. Volatiles were removed under reduced pressure and the residue was purified by silica gel chromatography (eluting with 0-10% MeOH) followed by reversed-phase chromatography (eluting with 0-100% MeCN / H2O) to give compound 121. 1 H NMR(400MHz,DMSO-d6)δ12.79(s,1H),8.04(s,1H),7.83(s,1H),6.80(s,1H),6 .78(s,1H),5.33(s,1H),4.49–4.40(m,3H),4.05–3.91(m,2H),3.77(d,J=11.4H z,1H),3.64(dd,J=11.5,3.1Hz,1H),3.49(td,J=11.7,2.9Hz,1H),3.23–3.11( m,1H),2.43–2.31(m,4H),1.81(dd,J=20.9,11.5Hz,4H),1.20(d,J=6.6Hz,3H).

[0807] Compound 125

[0808] Step 1. Under rt, di-tert-butyl dicarboxylate (2.65 g, 12.1 mmol) and DMAP (35 mg, 0.29 mmol) were added to a solution of 2-amino-3-bromopyridine (1.0 g, 5.8 mmol) in DCM (10 mL), followed by the slow addition of Et3N (1.8 mL, 12.9 mmol). The reaction mixture was stirred under rt for 18 h and then partitioned between water (50 mL) and DCM (40 mL). The aqueous layer was extracted with DCM (40 mL) and the combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated under vacuum. The residue was adsorbed onto silica gel for purification over 25 min by Combi-Flash (80 g Gold SiO2) (eluting from 100% hexane to 35% EtOAc / hexane) to give 1.8 g of (3-bromopyridine-2-yl)di-tert-butyl dicarboxylate as a colorless solid. MS(+ESI)m / z 395.1 / 397.1(M+Na)

[0809] Step 2. Di-tert-butyl (3-bromopyridin-2-yl)dicarbamate (150 mg, 0.40 mmol), bis(pinacolyl)diboron (204 mg, 0.80 mmol), and potassium acetate (120 mg, 1.21 mmol) were dissolved in anhydrous DMF (1 mL), followed by the dissolution of Pd(dppf)Cl2CH2Cl2 (33 mg, 0.04 mmol). The reaction mixture was purged with argon and then heated to 85 °C for 16 h. The mixture was diluted with EtOAc and filtered through a diatomaceous earth mat. The evaporator was evaporated to give (tert-butoxycarbonyl)(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)pyridin-2-yl)carbamate, which was used without further purification.

[0810] Step 3. Dissolve 2-[[3-[4-iodo-6-[(3R)-3-methylmorpholin-4-yl]pyrazolo[3,4-b]pyridin-1-yl]pyrazol-1-yl]methoxy]ethyltrimethylsilane (120 mg, 0.22 mmol), K3PO4 (142 mg, 0.66 mmol), Pd(dppfCl2)CH2Cl2 (9 mg, 0.011 mmol), and (tert-butoxycarbonyl)(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)pyridin-2-yl)tert-butyl carbamate (186 mg, 0.44 mmol) in anhydrous DMF (2 mL). Purge the reactants with argon and heat to 85 °C for 16 h. The product was purified for 20 min by combiflash (C18, 26 g) of 5-100% MeCN (0.1% formic acid) in H2O to give (R)-(tert-butoxycarbonyl)(3-(6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)pyridin-2-yl)carbamate (35 mg).

[0811] Step 4. Add TFA (0.40 mL, 5.2 mmol) and Et3SiH (0.03 mL, 0.17 mmol) to a solution of (R)-(tert-butyloxycarbonyl)(3-(6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)pyridin-2-yl)carbamate (35 mg, 0.05 mmol) in DCM (1.5 mL) and stir the reaction mixture for 1.5 h. The evaporator was evaporated and the residue was purified for 15 min by combi-flash (SiO2, 4 g) of hexane in 0-100% EtOAc to give (R)-3-(6-(3-methylmorpholino)-1-(1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)pyridine-2-amine (18 mg).

[0812] Compound 126

[0813] Step 1. Load (R)-4-(4-iodo-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-6-yl)-3-methylmorpholine (intermediate C, 700 mg, 1.30 mmol), (2-(N-(tert-butyl)aminosulfonyl)phenyl)boronic acid (492 mg, 1.68 mmol), 2M K₂CO₃ (2 mL, 4 mmol), Pd(PPh₃)₄ (75 mg, 0.065 mmol), and dioxane (7 mL) into a microwave tube. Seal the tube and flush with N₂ (vacuum / N₂, 3 cycles). Heat the mixture to 100 °C for 5 h. LCMS indicates the reaction is complete. After cooling to rt, dilute the mixture with EtOAc (40 mL) and water (40 mL). The layers were separated, and the aqueous layer was extracted with EtOAc (70 mL). The combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated to dryness. The residue was adsorbed onto silica gel for purification by ISCO CombiFlash (40 g column, SiO2 Gold) (eluting with 20–100% EtOAc / hexane) to give 700 mg (R)-N-(tert-butyl)-2-(6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)benzenesulfonamide.

[0814] Step 2. Under rt, TFA (9 mL, 118 mmol) was added to a solution of (R)-N-(tert-butyl)-2-(6-(3-methylmorpholino)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)benzenesulfonamide (700 mg, 1.12 mmol) and triethylsilane (0.68 mL, 4.26 mmol) in DCM (10 mL). The reaction mixture was stirred under rt for 4 h. The volatiles were removed under vacuum and the residue was dissolved in TFA (10 mL). The mixture was stirred under rt for 18 h, then heated to 40 °C for 1 h, and then heated to 50 °C for 1 h. The volatiles were removed under reduced pressure and the mixture was co-evaporated with DCM (3x). The residue was adsorbed onto silica gel for purification by ISCO CombiFlash (24 g column GoldSiO2) (eluting with 30-100% EtOAc / hexane). The desired product fraction was fractionated and concentrated to dryness under reduced pressure. The residue was dissolved in CH3CN (3 mL) and water (5 mL) for lyophilization to give 350 mg of (R)-2-(6-(3-methylmorpholino)-1-(1H-pyrazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-4-yl)benzenesulfonamide as a pale yellow foam. +ESI[M+1]: 440.2. Purity by HPLC at 254 nm: >99%, 10-90% CH3CN / H2O (+0.1% formic acid), over 20 min. 1 H NMR (400MHz, DMSO): δ12.83 (s; 1H); 8.11-8.13 (m; 1H); 7.86 (s; 1H); 7.68-7.72 ( m; 2H); 7.61 (s; 1H); 7.50-7.52 (m; 1H); 7.42 (s; 2H); 6.83 (s; 1H); 6.80 (s; 1H); 4. 33-4.38 (m; 1H); 4.08 (d; J=13.32Hz; 1H); 3.97-4.00 (m; 1H); 3.73-3.76 (m; 1H); 3.64-3.68 (m; 1H); 3.49-3.55 (m; 1H); 3.15-3.21 (m; 1H); 1.22 (d; J=6.61Hz; 3H).

[0815] Compound 138

[0816] Step 1. (3R)-4-[4-iodo-1-[2-[(4-methoxyphenyl)methyl]pyrazol-3-yl]pyrazolo[3,4-b]pyridin-6-yl]-3-methylmorpholine (500 mg, 0.943 mmol), bis(pinacol)diboron (359 mg, 1.41 mmol), and potassium acetate (324 mg, 3.30 mmol) were combined in DMF (5 mL) and the solution was degassed by sonicating for 10 min and then bubbling N2 into the mixture. Pd(dppf)Cl2·DCM (69 mg, 0.0943 mmol) was then added and the mixture was degassed again for 5 min. The reaction mixture was then heated to 95 °C for 2 h. The mixture was cooled to rt and partitioned between EtOAc and water (3 volumes each). The organic layer was washed with water (2 × 3 volumes), dried over Na2SO4, and concentrated to dryness. The product was then purified by combiflash (0-100% EtOAc / hex).

[0817] Step 2. To a solution of (3R)-4-[1-[2-[(4-methoxyphenyl)methyl]pyrazol-3-yl]-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)pyrazolo[3,4-b]pyridin-6-yl]-3-methylmorpholine (220 mg, 0.415 mmol) and 3-bromo-6-(trifluoromethyl)pyridin-2-amine (200 mg, 0.830 mmol) in DMF (9 mL), add an aqueous solution of K2CO3 (1.1 mL, 1.24 mmol), followed by the addition of the Pd(dppf)Cl2-DCM complex (68 mg, 0.083 mmol). The reactants were heated in a microwave to 110 °C and maintained for 10 min. The mixture was cooled to rt and partitioned between EtOAc and water (3 volumes each). The organic layer was washed with water (2 × 3 volumes), dried over Na₂SO₄, and concentrated to dryness. The product was then purified by combiflash (0-100% EtOAc / hex).

[0818] Step 3. Dissolve 3-[1-[2-[(4-methoxyphenyl)methyl]pyrazol-3-yl]-6-[(3R)-3-methylmorpholin-4-yl]pyrazolo[3,4-b]pyridin-4-yl]-6-(trifluoromethyl)pyridine-2-amine (120 mg, 0.213 mmol) in trifluoroacetic acid (3.0 mL, 0.21 mmol) and stir the reaction mixture for 2 h. Evaporate the reaction mixture to dryness, absorb the residue in DMSO (1 mL), and purify the product by reverse-phase combiflash (5-95% MeCN / water). +ESI[M+1]: 445.0. Purity by HPLC at 254 nm: >99%, 10-90% CH3CN / H2O (+0.1% formic acid), within 20 min. 1 H NMR(400MHz,DMSO-d6)δ12.81(s,1H),7.84(br m,2H),7.72(d,J=7.5Hz,1H),7.09(d,J=7.5Hz,1H),6.86–6.77(m,2H),6.45(s,2H),4.48(s,1H),4.11(d,J=13.4Hz,1H),4.07–3 .93(m,1H),3.75(d,J=11.3Hz,1H),3.64(d,J=9.8Hz,1H),3.49(t,J=11.1Hz,1H),3.20(t,J=12.6Hz,1H),1.20(d,J=6.9Hz,3H).

[0819] Compound 139

[0820] Step 1. To a solution of [1-[2-[(4-methoxyphenyl)methyl]pyrazol-3-yl]-6-[(3R)-3-methylmorpholin-4-yl]pyrazolo[3,4-b]pyridin-4-yl]boronic acid (250 mg, 0.558 mmol) and 3-bromo-6-methylpyridin-2-amine (0.33 mL, 1.12 mmol) in DMF (9 mL), K2CO3 (1.1 mL, 1.67 mmol) was added and the mixture was rinsed with nitrogen. Then, Pd(dppf)Cl2-DCM complex (91 mg, 0.11 mmol) was added. The reactants were heated in a microwave at 100 °C for 10 min, and the mixture was then cooled to rt and partitioned between EtOAc and water (3 volumes each). The organic layer was washed with water (2 × 3 volumes), dried over Na2SO4, and concentrated to dryness. The product was then purified by combiflash (0-100% EtOAc / hex).

[0821] Step 2. 3-[1-[2-[(4-methoxyphenyl)methyl]pyrazol-3-yl]-6-[(3R)-3-methylmorpholin-4-yl]pyrazolo[3,4-b]pyridin-4-yl]-6-methyl-pyridin-2-amine (250 mg, 0.490 mmol) was dissolved in trifluoroacetic acid (4.0 mL, 0.49 mmol), and the reaction mixture was heated to 60 °C and monitored by UPLC-MS. After 1 h, the reaction mixture was cooled to rt and evaporated to dryness the next day. The product was purified by reverse-phase combiflash (5-95% MeCN / water). +ESI[M+1]: 391.0. Purity by HPLC at 254 nm: >99%, 10-90% CH3CN / H2O (+0.1% formic acid), within 20 min. 1 HNMR(400MHz,DMSO-d6)δ12.81(s,1H),7.84(br,2H),7.59(br,1H),6.82–6.7 6(m,2H),6.68(s,1H),4.46(s,1H),4.09(d,J=13.3Hz,1H),3.97(d,J=10.2Hz ,1H),3.75(d,J=11.3Hz,1H),3.64(d,J=11.8Hz,1H),3.49(t,J=10.6Hz,1H), 3.19(t,J=13.0Hz,1H),2.55–2.50(m,3H),2.38(s,3H),1.22(d,J=6.6Hz,3H).

[0822] Compound 149

[0823] Step 1. Add N-chlorosuccinimide (170 mg, 1.27 mmol) to a solution of intermediate C (690 mg, 1.28 mmol) in chloroform (10 mL) and stir overnight at rt. Heat the solution to 65 °C for 1 h, then add another 138 mg of N-chlorosuccinimide and stir at 65 °C for 2 h. Remove the solvent under reduced pressure and purify the residue by silica gel chromatography (eluting with 0-100% EtOAc / hexane) to give 2-[[5-[5-chloro-4-iodo-6-[(3[R)-3-methylmorpholin-4-yl]pyrazolo[3,4-b]pyridin-1-yl]pyrazol-1-yl]methoxy]ethyl-trimethyl-silane (240 mg).

[0824] Step 2. Add K3PO4 (0.50 mL, 0.560 mmol) to a solution of 2-[[5-[5-chloro-4-iodo-6-[(3R)-3-methylmorpholin-4-yl]pyrazolo[3,4-b]pyridin-1-yl]pyrazol-1-yl]methoxy]ethyl-trimethyl-silane (107 mg, 0.187 mmol) and 4,4,5,5-tetramethyl-2-[2-(trifluoromethyl)phenyl]-1,3,2-dioxaborane (0.33 mL, 0.382 mmol) in 1,4-dioxane (1 mL). Rinse the vial with nitrogen, then add Pd(dppf)Cl2 (30 mg, 0.0373 mmol) and heat in a microwave at 110 °C for 3 h. Dilute the solution with water and DCM and filter on a phase separator. Wash the aqueous phase twice with DCM and evaporate the combined organic extracts under reduced pressure. The product was used in the next step without further purification.

[0825] Step 3. To the unpurified solution of 2-[[5-[5-chloro-6-[(3R)-3-methylmorpholin-4-yl]]-4-[2-(trifluoromethyl)phenyl]pyrazolo[3,4-b]pyridin-1-yl]pyrazol-1-yl]methoxy]ethyl-trimethylsilane (110 mg, 0.186 mmol) in DCM (1 mL), add 0.2 mL of triethylsilane and 1 mL of TFA. Stir the resulting solution at rt for 1 h and remove the solvent under reduced pressure. Purify the residue by reversed-phase chromatography (eluting with 0–100% MeCN / H2O), followed by further purification by normal-phase chromatography (eluting with 0–10% MeOH / DCM) to obtain the desired product as a 1:1 mixture of the transisomers, which is then used for bioassays. Further purification using chiral SFC yielded two isolated transisomers (4.0 mg, 6.8% and 4.7 mg, 8.0%, respectively). Mass spectrometry: m / z: 463.2.

[0826] Compound 150

[0827] Step 1. 2,6-Difluoro-4-iodo-pyridine-3-carboxaldehyde (9.00 g, 33.5 mmol) was dissolved in DMSO (330 mL), and (3R)-3-methylmorpholine (3.8 mL, 33.3 mmol) was added. The solution was heated at 120 °C for 2 h. The solution was cooled and added dropwise to water (1.5 L) with vigorous stirring. Ice was then added, and the suspension was stirred again for 2 h. The solid was filtered and dried under vacuum in a room for 15 h. The resulting beige solid (10.9 g) was dissolved in minimal DCM and purified by silica gel chromatography (0 to 100% EtOAc / hexane gradient) to give 2-fluoro-4-iodo-6-[(3R)-3-methylmorpholine-4-yl]pyridine-3-carboxaldehyde (6.36 g, 18.16 mmol) as a beige solid.

[0828] Step 2. Under reflux, 2-methyl-2-butene (82 mL, 164 mmol), sodium chlorite (7.56 g, 83.6 mmol), and sodium dihydrogen phosphate (2.64 g, 16.9 mmol) were added to a solution of 2-fluoro-4-iodo-6-[(3R)-3-methylmorpholin-4-yl]pyridine-3-carboxaldehyde (5.86 g, 16.7 mmol) in tert-butanol (20 mL) and water (6.5 mL). The resulting mixture was stirred under reflux for 15 h. A saturated aqueous solution of sodium sulfite was slowly added, followed by formic acid until an acidic pH was reached. EtOAc was added and the phases were separated. The aqueous phase was extracted three times with EtOAc, and the combined organic extracts were dried over MgSO4, filtered, and evaporated under reduced pressure to give a beige solid. This substance was ground in Et2O for 30 min and then filtered to give 2.06 g of a grayish-white solid. The filtrate was concentrated and purified using a 0 to 100% MeCN / water gradient on a 100 g C18 column to give an additional 2.83 g of beige solid, for a total of 4.89 g of 2-fluoro-4-iodo-6-[(3[R)-3-methylmorpholin-4-yl]pyridine-3-carboxylic acid.

[0829] Step 3. To a solution of 2-fluoro-4-iodo-6-[(3[R)-3-methylmorpholin-4-yl]pyridin-3-carboxylic acid (4.89 g, 13.4 mmol) in DMF (67 mL), add azinon-[2-[(4-methoxyphenyl)methyl]pyrazol-3-yl]ammonium dichloride (4.65 g, 16.0 mmol), followed by 2,6-dimethylpyridine (12 mL, 100 mmol). Then add HATU (6.17 g, 16.2 mmol) and stir the reaction mixture at rt for 1 h. Then, under vigorous stirring, add the solution dropwise to water (400 mL) to obtain a suspension, stir for 1 h, and then filter. The obtained solid was dried under vacuum for 15 h to obtain a beige solid, 2-fluoro-4-iodo-N'-[2-[(4-methoxyphenyl)methyl]pyrazol-3-yl]-6-[(3[R)-3-methylmorpholin-4-yl]pyridine-3-carbazide (7.48 g, 13.2 mmol).

[0830] Step 4. Dissolve 2-fluoro-4-iodo-N'-[2-[(4-methoxyphenyl)methyl]pyrazol-3-yl]-6-[(3R)-3-methylmorpholin-4-yl]pyridine-3-carbazide (2.00 g, 3.53 mmol) in DMF (70 mL) and add NaH (285 mg, 7.13 mmol). Stir the mixture at rt for 10 min, then slowly heat to 60 °C over 30 min. Add water, brine, and EtOAc and separate the phases. Extract the aqueous phase three times with EtOAc, and dry the combined organic matter over MgSO4, filter, and evaporate under reduced pressure. The obtained substance was purified by silica gel chromatography (0 to 10% MeOH / DCM gradient) to give 4-iodo-1-[2-[(4-methoxyphenyl)methyl]pyrazol-3-yl]-6-[(3R)-3-methylmorpholin-4-yl]-2H-pyrazolo[3,4-b]pyridin-3-one (1.19 g, 2.18 mmol) as a brown solid.

[0831] Step 5. Add K₂CO₃ (0.28 mL, 0.55 mmol) to a solution of 4-iodo-1-[2-[(4-methoxyphenyl)methyl]pyrazol-3-yl]-6-[(3R)-3-methylmorpholin-4-yl]-2H-pyrazolo[3,4-b]pyridin-3-one (100 mg, 0.183 mmol) and 2-trifluoromethylphenylboronic acid (87 mg, 0.43 mmol) in 1,4-dioxane (1.8 mL). Purge the mixture with nitrogen for 5 minutes, then add Pd(dppf)Cl₂ (30 mg, 0.037 mmol) and heat to 110 °C under microwave irradiation for 15 minutes. Add water and DCM, and separate the phases. The aqueous phase was extracted three times with DCM, and the organic extracts were combined, dried over MgSO4, filtered, and evaporated under reduced pressure to give 1-[2-[(4-methoxyphenyl)methyl]pyrazol-3-yl]-6-[(3[R)-3-methylmorpholin-4-yl]-4-[2-(trifluoromethyl)phenyl]-2H-pyrazolo[3,4-b]pyridin-3-one (103 mg, 0.182 mmol) as a dark solid. The unpurified substance was used in the next step.

[0832] Step 6. The unpurified substance from Step 5 was dissolved in trifluoroacetic acid (2.0 mL) and stirred at 60 °C for 1.5 h, then concentrated under vacuum. The resulting substance was dissolved in DMSO (1 mL) and purified by silica gel chromatography (0 to 100% MeCN / water gradient) to give 6-[(3R)-3-methylmorpholin-4-yl]-1-(1H-pyrazol-5-yl)-4-[2-(trifluoromethyl)phenyl]-2H-pyrazolo[3,4-b]pyridin-3-one (35 mg, 0.079 mmol) as a beige solid. 1 H NMR (400MHz, DMSO-d6) δ12.60(s,1H),10.86(s,1H),7.84(d,J=7.8Hz,1H),7.79–7.75(m,1H),7.7 3(t,J=7.5Hz,1H),7.65(t,J=7.6Hz,1H),7.47(d,J=7.0Hz,1H),6.76(s,1H),6.48(d,J=5.5Hz,1H) ,4.44–4.33(m,1H),4.12–4.03(m,1H),4.01–3.91(m,1H),3.73(d,J=11.4Hz,1H),3.65(dd,J=11. 4,2.7Hz,1H),3.57–3.46(m,1H),3.17(td,J=12.7,12.2,3.5Hz,1H),1.20(dd,J=10.9,6.6Hz,3H).

[0833] Example 2. ATR / ATRIP Enzymatic Assay

[0834] ATR kinase activity was detected using the AlphaScreen system to measure the phosphorylation of the substrate protein p53. Recombinant purified ATR / ATRIP (Eurofins catalog number 14-953) was mixed with serially diluted compounds in 10% DMSO to a final concentration of 0.63 nM in assay buffer (50 mM Hepes pH 7.4, 0.1 mM vanadate, 0.5 mM DTT, 0.1 mM EGTA, 5 mM MnCl2, 0.01% Brij-30, 1% glycerol, 0.05% BSA). The final DMSO concentration was 1.25%. A premixture of GST-tagged p53 (full length, Enzo Life Sciences catalog number BML-FW9370) and 5'-ATP (Sigma-Aldrich catalog number 10519979001, Roche Diagnostic) in the assay buffer was added to the enzyme:compound mixture to a final concentration of 25 nM GST-p53 and 3 μM ATP. The reaction was allowed to proceed at room temperature for 1 hour, then terminated by adding a premixed solution of phosphorylated p53 (Ser 15) antibody (New England Biolabs catalog 9284S) at a final dilution of 1:3000, glutathione donor beads (PerkinElmer Life Sciences catalog 6765301) and protein A receptor beads (PerkinElmer Life Sciences catalog 670137) at a final bead concentration of 14.3 μg / mL in buffer (60 mM EDTA in 50 mM Tris, pH 7.4, and 0.1% BSA). The plates were incubated in the dark at room temperature for 4 hours and read on a BMG Polarstar using an AlphaScreen-specific filter. Assays were performed using white polypropylene half-zone plates (Costar catalog 3693) in a 96-well format. IC50 50 The values ​​were determined using a 4-parameter fitting algorithm.

[0835] Example 3. ATR assay in HeLa cells

[0836] HeLa S3 cells were seeded in standard F-12K 10% FBS medium at a density of 16K cells per 25 μL well in 384-well plates and incubated overnight at 37°C and 5% CO2. The medium was then replaced with 20 μL of Opti-MEM (phenol red-free) per well, and 5 μL of serially diluted compound (DMSO) was added to each well. Cells and compound were incubated at room temperature for 20 min, followed by the addition of 5 μL of gemcitabine (final concentration 1.5 μM). The plates were incubated at 37°C and 5% CO2 for 3.5–4 h. The medium was removed, and cells were lysed in 15 μL of PerkinElmer lysis buffer for 10–20 min; then 4 μL of the lysate was transferred to a 384-well Proxi white plate (PerkinElmer Life Sciences catalog number 6008280). Quantification of CHK1 phosphorylation at Ser345 was performed using Alphascreen SureFire CHK1 p-Ser345 (PerkinElmer Life Sciences catalog number TGRCHK1S10K) and Alphascreen protein A (PerkinElmer Life Sciences catalog number 67060617C). Plates were read on Envision using AlphaScreen-specific filters. IC 50 The values ​​were determined using a 4-parameter fitting algorithm.

[0837] The exemplary prepared compounds and their activities in the ATR / ATRIP enzymatic assay are shown in Table 2.

[0838] Table 2

[0839]

[0840]

[0841]

[0842]

[0843]

[0844]

[0845]

[0846]

[0847]

[0848]

[0849]

[0850]

[0851]

[0852]

[0853] In Table 2, the method column indicates the above-mentioned methods for preparing the compounds.

[0854] Exemplary prepared compounds and their ATR inhibitory activity in HeLa S3 whole-cell assays are shown in Table 3 below.

[0855] Table 3

[0856]

[0857]

[0858]

[0859]

[0860]

[0861]

[0862]

[0863]

[0864]

[0865]

[0866]

[0867] In Table 3, the method column indicates the above-mentioned methods for preparing the compounds.

[0868] Other implementation plans

[0869] Various modifications and variations of the described invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. While the invention has been described in conjunction with specific embodiments, it should be understood that the claimed invention should not be unduly limited to those specific embodiments. Indeed, various modifications intended to carry out the described methods of the invention that will be apparent to those skilled in the art are within the scope of the invention.

[0870] Other embodiments are described in the claims.

Claims

1. A compound of formula (I): Or its pharmaceutically acceptable salt. in It is a double bond, and each Y is independently either N or CR. 4 ;or It is a single bond, and each Y is independently NR. Y carbonyl or C(R) Y )2; where each R Y Independently H or optionally substituted C 1-6 alkyl; R 1 C is an optional replacement 1-6 Alkyl or H; R 2 C is an optional replacement 2-9 Heterocyclic groups, optionally substituted C 1-6 Alkyl, optionally substituted C 3-8 cycloalkyl, optionally substituted C 2-9 Heterocyclic C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 Heteroaryl, optionally substituted C 1-9 heteroaryl C 1-6 Alkyl, halogen, –N(R) 5 2. –OR 5 –CON(R) 6 )2、–SO2N(R 6 )2、–SO2R 5A Or –Q–R 5B ; R 3 C is an optional replacement 1-9 Mixed aromatics; Each R 4 Independently hydrogen, halogen, or optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 alkenyl or optionally substituted C 2-6 alkynyl group; Each R 5 Independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 heteroaryl or –SO2R 5A ; or two Rs 5 Together with the atoms it is attached to, they combine to form optionally substituted C atoms. 2-9 Heterocyclic groups; Each R 5A C can be substituted independently. 1-6 Alkyl, optionally substituted C 3-8 cycloalkyl or optionally substituted C 6-10 aryl; R 5B Hydroxyl group, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 heteroaryl, –N(R) 5 )2、–CON(R 6 )2、–SO2N(R 6 )2、–SO2R 5A Or optional replacement of C 1-6 alkoxy; Each R 6 Independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkoxyalkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 3-8 cycloalkyl or optionally substituted C 1-9 heteroaryl; or two R 6 Together with the atoms it is attached to, they combine to form optionally substituted C atoms. 2-9 Heterocyclic groups; Q is an optional substitution for C. 2-9 Heterocyclic groups, optionally substituted C 3-8 Cycloalkylene, optionally substituted C 1-9 heteroaryl or optionally substituted C 6-10 Alpha-aryl; and X is hydrogen or halogen. Each heterocyclic group, sub-heterocyclic group, C 1-9 Heteroaryl and heteroaryl groups contain one to four heteroatoms independently selected from nitrogen, oxygen, and sulfur; Each optionally substituted alkyl, alkenyl, alkynyl, or alkoxy group may be substituted by one, two, or three independently selected groups from the following: –N(R 5 )2;-NH(C 3-6 cycloalkyl); C 6-10 Aryl; C 6-10 aryloxy group; azide group; C 3-8 cycloalkyl; C 3-8 Cycloalkoxy; C 3-16 Cycloalkenyl; C 3-16 Cycloalkynyl; Halogen; C 2-9 Heterocyclic group; (C 2-9 Heterocyclic group (oxygen group); C 1-9 heteroaryl; hydroxyl; cyano; C 1-6 alkylsulfonyl; C 1-6 alkylsulfinyl; C 1-6 Alkyl sulfide group; =O; =S; -SO2R a , where R a It is -NH2, -NH(C 1-6 alkyl), -NH(C 3-6 cycloalkyl) or C 3-8 cycloalkyl; and =NR', where R' is H, C 1-6 Alkyl, C 6-10 Aryl and C 2-9 Heterocyclic groups, where each C 2-9 Heterocyclic groups and C 1-9 Heteroaryl groups contain one to four heteroatoms independently selected from nitrogen, oxygen, and sulfur; Each optionally substituted cycloalkyl or cyclohexane may be replaced by one, two, or three independently selected groups from the following groups: C 1-6 Alkyl; C 2-6 alkenyl; C 2-6 alkynyl group; C 1-6 Alkoxy; C 1-6 alkylsulfinyl; C 1-6 alkyl sulfide; C 1-6 alkylsulfonyl; -NH2; -NH(C 1-6 Alkyl); -NH(C 3-6 cycloalkyl); C 6-10 Aryl; C 6-10 aryloxy group; azide group; C 3-8 cycloalkyl; C 3-8 Cycloalkoxy; C 3-16 Cycloalkenyl; C 3-16 Cycloalkynyl; halogen; a C group consisting of one or two heteroatoms selected from nitrogen, oxygen, and sulfur, spaced once, twice, three times, or four times independently of each other. 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 alkynyl group; C 2-9 Heterocyclic group; (C 2-9 Heterocyclic group (oxygen group); C 1-9 Heteroaryl; hydroxyl; cyano; =O; =S; -SO2R, where R is -NH2, -NH(C 1-6 alkyl), -NH(C 3-6 cycloalkyl) or C 3-8 cycloalkyl; =NR', where R' is H, C 1-6 Alkyl, C 6-10 Aryl or C 2-9 Heterocyclic groups; and -CON(R) A )2, where each R A Independently H or C 1-6 Alkyl, or two R A Together with the atoms they are attached to, they combine to form C. 2-9 Heterocyclic groups, where each C 2-9 Heterocyclic groups and C 1-9 Heteroaryl groups contain one to four heteroatoms independently selected from nitrogen, oxygen, and sulfur; Each of the optionally substituted heterocyclic group, sub-heterocyclic group, and C 1-9 heteroaryl and C 1-9 The heteroaryl group can be substituted by one, two, three, four, or five groups independently selected from the following groups: C 1-6 Alkyl; C 2-6 alkenyl; C 2-6 alkynyl group; C 1-6 Alkoxy; C 1-6 alkylsulfinyl; C 1-6 alkyl sulfide group; C 1-6 alkylsulfonyl; NH2; -NH(C 1-6 Alkyl); -NH(C 3-6 cycloalkyl); C 6-10 Aryl; C 6-10 aryloxy group; azide group; C 3-8 cycloalkyl; C 3-8 Cycloalkoxy; C 3-16 Cycloalkenyl; C 3-16 Cycloalkynyl; halogen; a C group consisting of one or two heteroatoms selected from nitrogen, oxygen, and sulfur, spaced once, twice, three times, or four times independently of each other. 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 alkynyl group; C 2-9 Heterocyclic group; (C 2-9 Heterocyclic group (oxy group); hydroxy group; cyano group; =O; =S; and =NR', where R' is H, C 1-6 Alkyl, C 6-10 Aryl or C 2-9 Heterocyclic groups, where each C 2-9 The heterocyclic group contains one to four heteroatoms independently selected from nitrogen, oxygen, and sulfur; and Each optionally substituted aryl or arylene group may be substituted by one, two, three, four, or five or more groups independently selected from the following groups: C 1-6 Alkyl; C 2-6 alkenyl; C 2-6 alkynyl group; C 1-6 Alkoxy; C 1-6 alkylsulfinyl; C 1-6 alkyl sulfide group; C 1-6 alkylsulfonyl; -NH2; -NH(C 1-6 Alkyl); -NH(C 3-6 cycloalkyl); C 6-10 Aryl; C 6-10 aryloxy group; azide group; C 3-8 cycloalkyl; C 3-8 Cycloalkoxy; C 3-16 Cycloalkenyl; C 3-16 Cycloalkynyl; halogen; C separated by one or two heteroatoms once, twice independently by one or two heteroatoms selected from nitrogen, oxygen, and sulfur, three times independently by one or two heteroatoms selected from nitrogen, oxygen, and sulfur, or four times independently by one or two heteroatoms selected from nitrogen, oxygen, and sulfur. 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 alkynyl group; C 2-9 Heterocyclic group; (C 2-9 Heterocyclic group (oxy group); hydroxy group; and cyano group, wherein each C 2-9 The heterocyclic group contains one to four heteroatoms independently selected from nitrogen, oxygen and sulfur.

2. The compound of claim 1, wherein the compound is a compound of formula (II): Or its pharmaceutically acceptable salt. in Each Y is independently either N or CR 4 ; R 1 C is an optional replacement 1-6 Alkyl or H; R 2 C is an optional replacement 2-9 Heterocyclic groups, optionally substituted C 1-6 Alkyl, optionally substituted C 3-8 cycloalkyl, optionally substituted C 2-9 Heterocyclic C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 Heteroaryl, optionally substituted C 1-9 heteroaryl C 1-6 Alkyl, halogen, –N(R) 5 2. –OR 5 –CON(R) 6 )2、–SO2N(R 6 )2、–SO2R 5A Or –Q–R 5B ; R 3 C is an optional replacement 1-9 Mixed aromatics; Each R 4 Independently hydrogen, halogen, or optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 alkenyl or optionally substituted C 2-6 alkynyl group; Each R 5 Independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 heteroaryl or –SO2R 5A ; or two Rs 5 Together with the atoms it is attached to, they combine to form optionally substituted C atoms. 2-9 Heterocyclic groups; Each R 5A C can be substituted independently. 1-6 Alkyl, optionally substituted C 3-8 cycloalkyl or optionally substituted C 6-10 aryl; R 5B Hydroxyl group, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 heteroaryl, –N(R) 5 )2、–CON(R 6 )2、–SO2N(R 6 )2、–SO2R 5A Or optionally substituted alkoxy groups; Each R 6 Independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkoxyalkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 aryl, optionally substituted C 3-8 cycloalkyl or optionally substituted C 1-9 heteroaryl; or two R 6 Together with the atoms it is attached to, they combine to form optionally substituted C atoms. 2-9 Heterocyclic groups; Q is an optional substitution for C. 2-9 Heterocyclic groups, optionally substituted C 3-8 Cycloalkylene, optionally substituted C 1-9 heteroaryl or optionally substituted C 6-10 Alpha-aryl; and X is hydrogen or halogen.

3. The compound of claim 1, wherein the compound is a compound of formula (IB): Or its pharmaceutically acceptable salt.

4. The compound of claim 1, wherein the compound is a compound of formula (IB-a): Or its pharmaceutically acceptable salt.

5. The compound of claim 1, wherein the compound is a compound of formula (Ia): Or its pharmaceutically acceptable salt.

6. The compound of claim 1, wherein the compound is a compound of formula (IA): Or its pharmaceutically acceptable salt.

7. The compound of claim 1, wherein the compound is a compound of formula (IA-a): Or its pharmaceutically acceptable salt.

8. The compound of claim 1, wherein the compound is a compound of formula (IC): Or its pharmaceutically acceptable salt.

9. The compound of claim 1, wherein the compound is a compound of formula (IC-a): Or its pharmaceutically acceptable salt.

10. The compound of claim 1, wherein R 1 It is a methyl group.

11. The compound of claim 1, wherein R 2 C is an optional replacement 2-9 Heterocyclic groups, optionally substituted C 1-6 Alkyl, optionally substituted C 3-8 cycloalkyl, optionally substituted C 6-10 aryl, optionally substituted C 1-9 Heteroaryl, optionally substituted C 1-9 heteroaryl C 1-6 Alkyl, –N(R) 5 )2、–CON(R 6 )2、–SO2N(R 6 )2 or –SO2R 5A .

12. The compound of claim 1, wherein each R 5A C can be substituted independently. 1-6 Alkyl or optionally substituted C 3-8 Cycloalkyl.

13. The compound of claim 1, wherein each R 6 Independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 aryl or optionally substituted C 1-9 heteroaryl; or two R 6 Together with the atoms it is attached to, they combine to form optionally substituted C atoms. 2-9 Heterocyclic group.

14. The compound of claim 1, wherein R 2 for: –I、–SO2Me、 –SO2Ph、 –OMe、 –OCH2CF3、 15. The compound of claim 1, wherein R 3 A monocyclic C with optional substitution containing at least one nitrogen atom 1-9 Mixed aromatic compounds.

16. The compound of claim 1, wherein R 3 for:

17. The compound of claim 1, wherein R 4 It is hydrogen.

18. The compound of claim 1, wherein X is hydrogen.

19. A compound selected from the following compounds: And its pharmaceutically acceptable salts.

20. A pharmaceutical composition comprising the compound as described in any one of claims 1 to 19 and a pharmaceutically acceptable excipient.

21. Use of the compound of any one of claims 1 to 19 in the preparation of a medicament for inhibiting ATR kinase in cells expressing ATR kinase.

22. The use as claimed in claim 21, wherein the cells are in the subject.

23. Use of the compound according to any one of claims 1 to 19 in the preparation of a medicament for treating diseases or symptoms of excessive cell proliferation.

24. The use as described in claim 23, wherein the disease or symptom is cancer.

25. The use as described in claim 24, wherein the cancer is epithelial carcinoma, sarcoma, adenocarcinoma, leukemia, or melanoma.

26. The use as described in claim 24, wherein the cancer is an epithelial carcinoma selected from the group consisting of: thyroid brain-like epithelial carcinoma, familial thyroid brain-like epithelial carcinoma, acinar epithelial carcinoma, acinar epithelial carcinoma, adenoid cystic epithelial carcinoma, adenoid cystic epithelial carcinoma, adenomatous epithelial carcinoma, adrenocortical epithelial carcinoma, alveolar epithelial carcinoma, alveolar cell epithelial carcinoma, basal cell epithelial carcinoma, basal cell carcinoma, basaloid epithelial carcinoma, basal squamous cell epithelial carcinoma, bronchioloalveolar epithelial carcinoma, bronchiolar epithelial carcinoma, bronchial epithelial carcinoma, medullary epithelial carcinoma, bile duct cell epithelial carcinoma, choriocarcinoma. Skin carcinoma, colloid carcinoma, comedo-like carcinoma, main carcinoma, cribriform carcinoma, armored carcinoma, cutaneous carcinoma, columnar carcinoma, columnar cell carcinoma, ductal carcinoma, sclerothelial carcinoma, embryonal carcinoma, cerebral carcinoma, epidermoid carcinoma, adenoid carcinoma, exophytic carcinoma, ulcerative carcinoma, fibrocarcinoma, colloid carcinoma, gelatinous carcinoma, giant cell carcinoma, giant cell carcinoma, adenocarcinoma, granular cell carcinoma, pilostromal carcinoma, polycythian carcinoma, hepatocellular carcinoma, Schulte's cell carcinoma, hyaline carcinoma, adrenal carcinoma Epithelial carcinoma, infantile embryonic epithelial carcinoma, epithelial carcinoma in situ, intraepithelial epithelial carcinoma, intraepithelial intraepithelial carcinoma, Klinefelter's cell carcinoma, Kurtschitzky cell carcinoma, large cell epithelial carcinoma, lenticular epithelial carcinoma, lenticular epithelial carcinoma, lipomatous epithelial carcinoma, lymphoepithelial carcinoma, medullary epithelial carcinoma, medullary epithelial carcinoma, melanocyte epithelial carcinoma, soft epithelial carcinoma, mucinous epithelial carcinoma, mucinous cell epithelial carcinoma, mucinous epithelial carcinoma, mucinous epithelial carcinoma, myxomatous epithelial carcinoma, nasopharyngeal epithelial carcinoma, oat cell epithelial carcinoma, ossifying epithelial carcinoma, osteoid epithelial carcinoma, papillary epithelial carcinoma, periportal epithelial carcinoma Skin carcinoma, invasive preepithelial carcinoma, acanthosis nigra, soft pasty carcinoma, renal cell carcinoma, reservoir cell carcinoma, sarcomatoid carcinoma, Schneider's carcinoma, sclerosing carcinoma, scrotal carcinoma, signet ring cell carcinoma, simple carcinoma, small cell carcinoma, potato-shaped carcinoma, spherical cell carcinoma, spindle cell carcinoma, cavernous carcinoma, squamous cell carcinoma, string carcinoma, capillary carcinoma, angiotensinus carcinoma, transitional cell carcinoma, nodular carcinoma, nodular epithelial carcinoma, verrucous carcinoma, and villous carcinoma.

27. The use as claimed in claim 24, wherein the cancer is a sarcoma selected from the group consisting of: chondrosarcoma, fibrosarcoma, lymphosarcoma, melanosarcoma, myxosarcoma, osteosarcoma, Ebernathy's sarcoma, liposarcoma, liposarcoma, alveolar soft tissue sarcoma, ameloblastic sarcoma, botryoid sarcoma, chloroma sarcoma, choriocarcinoma, embryonal sarcoma, Williams' tumor sarcoma, endometrial sarcoma, stromal sarcoma, Ewing's sarcoma, and fasciitis. Membranous sarcoma, fibroblastic sarcoma, giant cell sarcoma, granulocyte sarcoma, Hodgkin's sarcoma, idiopathic multiple pigmented hemorrhagic sarcoma, B-cell immunoblastic sarcoma, T-cell immunoblastic sarcoma, Jensen's sarcoma, Kaposi's sarcoma, Kupffer cell sarcoma, angiosarcoma, leukemic sarcoma, malignant mesenchymal sarcoma, paraosteal sarcoma, reticulum cell sarcoma, Rouss's sarcoma, serous cystic sarcoma, synovial sarcoma, and capillary hemangiosarcoma.

28. The use as claimed in claim 24, wherein the cancer is a leukemia selected from the group consisting of: non-lymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, acute promyelocytic leukemia, adult T-cell leukemia, non-leukemic leukemia, leukemic leukemia, basophilic leukemia, blastic leukemia, bovine leukemia, chronic myeloid leukemia, cutaneous leukemia, embryonic leukemia, eosinophilic leukemia, Gross's leukemia, hairy cell leukemia, hematopoietic leukemia, hematopoietic leukemia, histiocytic leukemia, stem cell leukemia, acute monocytic leukemia. Leukemia, leukopenic leukemia, lymphocytic leukemia, lymphoblastic leukemia, lymphocytic leukemia, lymphoid leukemia, lymphosarcoma cell leukemia, mast cell leukemia, megakaryocytic leukemia, micromyeloid leukemia, monocytic leukemia, myeloblastic leukemia, myeloid leukemia, myeloid granulocytic leukemia, granulocytic-monocytic leukemia, Neghly leukemia, plasma cell leukemia, multiple myeloma, plasma cell leukemia, promyelocytic leukemia, Riddle cell leukemia, Schilling leukemia, stem cell leukemia, subleukemic leukemia, and undifferentiated cell leukemia.

29. The use as claimed in claim 24, wherein the cancer is a melanoma selected from the group consisting of: acral lentigines melanoma, amelanoma, benign juvenile melanoma, Claudemann's melanoma, S91 melanoma, Harpa's II melanoma, juvenile melanoma, malignant lentigines melanoma, malignant melanoma, nodular melanoma, subungual melanoma, and superficial diffuse melanoma.

30. The use as described in claim 24, wherein the cancer is prostate cancer, thyroid cancer, endocrine system cancer, brain cancer, breast cancer, cervical cancer, colon cancer, head and neck cancer, liver cancer, kidney cancer, lung cancer, non-small cell lung cancer, melanoma, mesothelioma, ovarian cancer, sarcoma, gastric cancer, uterine cancer, medulloblastoma, ampullary cancer, colorectal cancer, or pancreatic cancer.

31. The use as described in claim 24, wherein the cancer is Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, neuroblastoma, glioma, glioblastoma multiforme, ovarian cancer, rhabdomyosarcoma, primary thrombocytosis, primary macroglobulinemia, primary brain tumor, cancer, malignant pancreatic insulinoma, malignant carcinoid tumor, bladder cancer, precancerous skin lesions, testicular cancer, lymphoma, thyroid cancer, neuroblastoma, esophageal cancer, urogenital tract cancer, malignant hypercalcemia, endometrial cancer, adrenocortical cancer, endocrine and exocrine pancreatic growths, medullary thyroid carcinoma, thyroid brain-like epithelial carcinoma, melanoma, colorectal cancer, papillary thyroid carcinoma, hepatocellular carcinoma, or prostate cancer.

32. The use as described in claim 23, wherein the subject is suffering from pre-malignant symptoms and requires treatment.

Images