Aza-heterobicyclic inhibitors of mat2a and methods of use for treating cancer

Abstract

To provide aza-heterobicyclic inhibitors of MAT2A and methods of use for treating cancer.SOLUTION: The present disclosure provides compounds according to general formula I and their pharmaceutically acceptable salts, tautomers, and / or isotopologues, as described in the disclosure. The compounds are inhibitors of methionine adenosyltransferase isoform 2A (MAT2A). Also provided are pharmaceutical compositions and methods of using the compounds for treating cancers, including some cancers in which the gene encoding methylthioadenosine phosphorylase (MTAP) is deleted.SELECTED DRAWING: None

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 62 / 785,519, filed December 27, 2018, which is incorporated by reference herein in its entirety. [Background technology]

[0002] Methionine adenosyltransferase (MAT), also known as S-adenosylmethionine synthetase, is a cellular enzyme that catalyzes the synthesis of S-adenosylmethionine (SAM, or ADOMET) from methionine and ATP, which is thought to be the rate-limiting step of the methionine cycle. SAM is a propylamino donor in polyamine biosynthesis, a major methyl donor for DNA methylation, and is involved in gene transcription, cell proliferation, and the production of secondary metabolites.

[0003] Two genes, designated MAT1A and MAT2A, encode two distinct catalytic MAT isoforms. A third gene, MAT2B, encodes the MAT2A regulatory subunit. MAT1A is specifically expressed in the adult liver, while MAT2A is widely distributed. MAT isoforms Because the MAT1A-expressing cells have different catalytic and regulatory properties, MAT1A-expressing cells have significantly higher SAM levels than MAT2A-expressing cells. MAT2A promoter hypomethylation and histone acetylation have been shown to increase MAT2A expression. See, for example, M. Vazquez-Chantada et al., Gastroenterology 138 (2010) 1943-53; M. Frau et al., J. Hepatol. 59 (2013) 830-41; M. Frau et al., Hepatology 56 (2012) 165-75; and RM Pascale et al., Transl. Gastroenterol. Hepatol. 3 (2018) 36.

[0004] In hepatocellular carcinoma (HCC), decreased expression of MAT1A and increased expression of MAT2A occur, known as the MAT1A:MAT2A switch. This switch, accompanied by increased expression of MAT2B, leads to lower SAM content, which provides a growth advantage to hepatoma cells. MAT2A plays an important role in promoting hepatoma cell proliferation and is therefore a target for antitumor therapy. Recent studies have shown that silencing using small interfering RNAs substantially suppresses proliferation and promotes growth in hepatoma cells. It has been shown to induce apoptosis in mice. See, for example, T. Li et al., J. Cancer 7(10) (2016) 1317-1327.

[0005] Some cancer cell lines that are MTAP-deficient are particularly sensitive to MAT2A inhibition. Marjon et al. (Cell Reports 15(3) (2016) 574-587). MTAP (methylthioadenosine phosphorylase) is an enzyme widely expressed in normal tissues that catalyzes the conversion of methylthioadenosine (MTA) to adenine and 5-methylthioribose-1-phosphate. Adenine is recycled to generate adenosine monophosphate, and 5-methylthioribose-1-phosphate is converted to methionine and formate. Because of this salvage pathway, MTA can serve as an alternative purine source when de novo purine synthesis is blocked, for example, with antimetabolites such as L-alanosine.

[0006] MAT2A is dysregulated in additional cancers lacking MTAP deletion, including hepatocellular carcinoma and leukemia. J. Cai et al., Cancer Res. 58 (1998) 1444-1450; T. S. Jani et al., Cell. Res. 19 (2009) 358-369. Silencing MAT2A expression via RNA interference results in antiproliferative effects in several cancer models. H. Chen et al., Gastroenterology 133 (2007) 207-218; Q. Liu et al. Hepatol. Res. 37 (2007) 376-388.

[0007] Many human and mouse malignant cells lack MTAP activity. MTAP deficiency is not only present in tissue culture cells, but also in primary leukemia, glioma, melanoma, pancreatic cancer, non-small cell lung cancer (NSCLC), bladder cancer, astrocytoma, osteosarcoma, head and neck cancer, myxoid chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, soft tissue sarcoma, non-Hodgkin's lymphoma, and mesothelioma. The gene encoding human MTAP is located on human chromosome 9p in region 9p21. This region also contains the tumor suppressor genes p16INK4A (also known as CDKN2A) and p15INK4B. These genes encode p16 and p15, inhibitors of the cyclin D-dependent kinases cdk4 and cdk6, respectively. Alternatively, the p16INK4A transcript can be an alternative reading frame (ARF) inserted into a transcript encoding p14ARF. p14ARF binds to MDM2 and prevents p53 degradation (Pomerantz et al. (1998) Cell 92:713-723). The 9p21 chromosomal region is of interest because it is frequently homozygously deleted in a variety of cancers, including leukemia, non-small cell lung cancer, pancreatic cancer, glioma, melanoma, and mesothelioma. Deletions often inactivate multiple genes. For example, Cairns et al. ((1995) Nat. Gen. 11:210-212), after studying over 500 primary tumors, found that nearly all deletions identified in the tumors were homozygous deletions. reported that the MTAP gene, pl4ARF, and p16INK4A, comprised a 170-kb region. Carson et al. (WO 99 / 67634) reported a correlation between tumor development stage and homozygosity loss of the MTAP-encoding gene and the p16-encoding gene. For example, deletion of the MTAP gene, but not p16INK4A, was reported to be indicative of cancer at an early stage of tumor development, whereas deletion of the p16 and MTAP-encoding genes was reported to be indicative of cancer at a more advanced stage of tumor development. In some osteosarcoma patients, the MTAP gene was present at the time of diagnosis but was deleted at later time points (Garcia-Castellano et al., Clin. Cancer Res. 8(3) 2002 782-787). [Prior art documents] [Non-patent literature]

[0008] [Non-Patent Document 1] M. Vazquez-Chantada et al., Gastroenterology 138 (2010) 1943-53 [Non-patent document 2] M. Frau et al., J. Hepatol. 59 (2013) 830-41 [Non-patent document 3] M. Frau et al., Hepatology 56 (2012) 165-75 [Non-patent document 4] RM Pascale et al., Transl. Gastroenterol. Hepatol. 3 (2018) 36 [Non-patent document 5] T. Li et al., J. Cancer 7(10) (2016) 1317-1327 [Non-patent document 6] Marjon et al. (Cell Reports 15(3) (2016) 574-587) [Non-Patent Document 7] J. Cai et al., Cancer Res. 58 (1998) 1444-1450 [Non-patent document 8] TS Jani et al., Cell. Res. 19 (2009) 358-369 [Non-Patent Document 9] H. Chen et al., Gastroenterology 133 (2007) 207-218 [Non-Patent Document 10] Q. Liu et al. Hepatol. Res. 37 (2007) 376-388 [Non-Patent Document 11] Pomerantz et al. (1998) Cell 92:713-723 [Non-Patent Document 12] Garcia-Castellano et al., Clin. Cancer Res. 8(3) 2002 782-787 Summary of the Invention [Means for solving the problem]

[0009] The present disclosure provides compounds that inhibit MAT2A. The compounds and pharmaceutical compositions thereof are They are useful in methods of treating a variety of cancers, including those that are refractory to standard treatments such as surgery, radiation therapy, chemotherapy, and hormone therapy.

[0010] Thus, according to some embodiments, the present disclosure provides compounds according to the following general formula I, or pharmaceutically acceptable salts, tautomers, and / or isotopologues thereof: [ka]

[0011] In general formula I, L is O, S, NR, or a bond. R is H or C1-C6 alkyl.

[0012] R 1 is selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C3-C6 carbocyclyl, -(C1-C6 alkyl)(C3-C6 carbocyclyl), and -(C1-C6 alkyl)(C3-C6 cycloalkenyl); wherein R 1 Any alkyl in R is linear or branched. 1 is optionally substituted by 1 to 6 halo or 1 to 6 deuterium.

[0013] Alternatively, in one embodiment, when L is NR, R and R in combination with L 1 is one One or more R A and represents a 3- to 6-membered heterocycloalkyl optionally substituted with , wherein 1-4 ring members are independently selected from N, O, and S.

[0014] R 2 and R 3 is C2-C6 alkynyl, C6-C 10 Aryl, C3-C6 carbocyclyl, 5-10 membered Heteroaryl (wherein 1 to 4 members of the heteroaryl are independently selected from N, O, and S) and 3- to 14-membered heterocycloalkyl (wherein 1 to 4 of the heterocycloalkyl R 2 Oh BiR 3 is R A , OR A , halo, -N=NR A , N.R. A R B , -(C1-C6 alkyl)NR A R B , -C(O)OR A , -C(O)NR A R B , -OC(O)R A , —SI(C1-C6 alkyl)3, and —CN; or It is independently and optionally substituted by multiple substituents.

[0015] R 4 is H, C1-C6 alkyl (optionally one or more halo, hydroxy, or 3-14 membered heterocycloalkoxy (wherein 1-4 members of the heterocycloalkoxy are independently selected from N, O, and S)), —O(C1-C 6- alkyl) (optionally substituted with one or more halo -OH, halo, -CN, -(C1-C6 alkyl)NR A R B , and -NR A R B Selected from the group consisting of can be.

[0016] R 5 is H, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, Halo, -CN, and -NR C R D is selected from the group consisting of:

[0017] R A and R Bis H, -CN, -hydroxy, oxo, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, -NH2, -S(O) 0-2 -(C1-C6 alkyl), -S(O) 0-2 -(C6-C 10 aryl), -C(O)(C1-C6 alkyl), -C(O)(C3-C 14 carbocyclyl), -C3-C 14 Carbosic -(C1-C6 alkyl)(C3-C 14 carbocyclyl), C6-C 10 Aryl, 3-14 membered heterocyclic -(C1-C6 alkyl)-(3-14 membered heterocycloalkyl), where 1-4 members of the heterocycloalkyl are independently selected from N, O, and S; and -(C1-C6 alkyl)-(3-14 membered heterocycloalkyl), where 1-4 members of the heterocycloalkyl are independently selected from N, O, and S; and -(C1-C6 alkyl)-(3-14 membered heterocycloalkyl), where 1-4 members of the heteroaryl are independently selected from N, O, and S. are independently selected from the group consisting of:

[0018] R A and R B Each alkyl, alkoxy, alkenyl, alkynyl, aryl, carbocyclyl, heterocycloalkyl, and heteroaryl moiety in 10 and independently selected from the group consisting of aryl, 3-14 membered heterocycloalkyl, and -(C1-C6 alkyl)-(3-14 membered heterocycloalkyl) (wherein 1-4 members of the ring are independently selected from N, O, and S), and 5-10 membered heteroaryl (wherein 1-4 members of the heteroaryl are independently selected from N, O, and S), -NHC(O)(O-C1-C6 alkyl), -NO2, -CN, oxo, -C(O)OH, -C(O)O(C1-C6 alkyl), -C1-C6 alkyl(C1-C6 alkoxy). Si), -C(O)NH2, C1-C6 alkyl, -C(O)C1-C6 alkyl, -OC1-C6 alkyl, -Si(C1-C6 alkyl)3, -S(O) 0-2 -(C1-C6 alkyl), C6-C 10 Aryl, -(C1-C6 alkyl)(C6-C 10 Ally -(C-C alkyl), 3- to 14-membered heterocycloalkyl, and -(C-C alkyl)-(3- to 14-membered heterocycle), where 1-4 members of the heterocycle are independently selected from N, O, and S, and -O(C-C 14 R is optionally substituted with one or more substituents selected from the group consisting of aryl. A and R B Each alkyl, alkenyl, aryl, and heterocycloalkyl substituent may be deuterium. , hydroxy, -OC1-C6 alkyl, halo, -NH2, -(C1-C6 alkyl)NH2, -C(O)OH, CN, and and oxo.

[0019] R C and R D are each independently selected from H and C1-C6 alkyl.

[0020] The present disclosure provides, in another embodiment, a therapeutically effective amount of a compound described herein, or a pharmaceutically acceptable salt, tautomer, and / or isotopologue thereof, as well as a pharmaceutically acceptable salt, tautomer, and / or isotopologue thereof. A pharmaceutical composition is provided comprising a suitable carrier.

[0021] In another embodiment, the disclosure provides a method of treating cancer in a subject suffering from cancer, comprising administering to the subject an effective amount of a MAT2A inhibitor compound described herein. to provide.

[0022] Yet another embodiment of the present disclosure provides a method for the synthesis of S-adenosylmethionine (SAM) in cells. 10. A method of inhibiting a cellular phenotype comprising introducing into a cell an effective amount of a compound described herein or a pharmaceutically acceptable salt, tautomer, and / or isotopologue thereof. is.

[0023] In another embodiment, the present disclosure provides a method for inhibiting the synthesis of S-adenosylmethionine (SAM) in a subject. 2. A method for inhibiting the growth of a tumor in a subject, the method comprising administering to the subject an effective amount of at least one compound described herein or a salt, tautomer, and / or isotope thereof. Regarding.

[0024] In one embodiment, the disclosure provides a method of treating cancer in a subject suffering from cancer, wherein the cancer has decreased or no methylthioadenosine phosphorylase (MTAP) gene expression compared to a cancer in which the MTAP gene or MTAP protein is present and / or fully functional. or absence of the MTAP gene, or reduced function of the MTAP protein, comprising administering to a subject a therapeutically effective amount of a compound described herein, or a pharmaceutically acceptable salt, tautomer, and / or isotopologue thereof. A method is provided.

[0025] The present disclosure also provides, in another embodiment, a compound for inhibiting the synthesis of S-adenosylmethionine (SAM). The compounds described herein or pharmaceutically acceptable salts, tautomers, and / or Or provide isotopologues.

[0026] In still further embodiments, the present disclosure provides a compound described herein or a pharmaceutically acceptable salt, tautomer, and / or isotopologue thereof for use in treating cancer in a subject suffering from cancer.

[0027] The present disclosure also provides the use of a compound described herein, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating cancer. In one embodiment, for example, the following items are provided: (Item 1) A compound according to the following general formula I: [ka] During the ceremony, L is O, S, NR, or a bond; R is H or C1-C6 alkyl; R 1 is selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C3-C6 carbocyclyl, -(C1-C6 alkyl)(C3-C6 carbocyclyl), and -(C1-C6 alkyl)(C3-C6 cycloalkenyl); is selected, wherein: R 1 Any alkyl in is linear or branched; R 1 is optionally substituted by 1 to 6 halo or 1 to 6 deuterium; or, if L is NR, R and R in combination with L 1 is one or more R A Replace with represents an optionally substituted 3- to 6-membered heterocycloalkyl, where 1-4 ring members are independently selected from N, O, and S; R 2 and R 3 is (C2-C6)alkynyl, C6-C 10 Aryl, C3-C6 carbocyclyl, 5-10 membered heteroaryl, wherein 1 to 4 members of the heteroaryl are independently selected from N, O, and S; and 3- to 14-membered heterocycloalkyl, wherein 1-4 members of the heterocycloalkyl are independently selected from N, O, and S; In the formula, R 2 and R3 is R A , OR A , halo, -N=NR A , N.R. A R B , -(C1-C6 alkyl)NR A R B , -C(O)OR A , -C(O)NR A R B , -OC(O)R A , —Si(C1-C6 alkyl)3, and —CN; is independently and optionally substituted by one or more substituents R 4 is H, C1-C6 alkyl (optionally substituted with one or more halo, hydroxy, or 3-14 membered heterocycloalkoxy (wherein 1-4 members of the heterocycloalkoxy are independently selected from N, O, and S)), —O(C1-C 6- alkyl) (with one or more halo (optionally substituted with), —OH, halo, —CN, —(C1-C6 alkyl)NR A R B , and -NR A R B from is selected from the group consisting of R 5 is H, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, ha -CN, and -NR C R D is selected from the group consisting of R A and R B is H, -CN, -hydroxy, oxo, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, -NH2, -S(O) 0-2 -(C1-C6 alkyl), -S(O) 0-2 -(C6-C 10 aryl), -C(O)(C1-C6 alkyl), -C(O)(C3-C 14 carbocyclyl), -C3-C14 Carbosic -(C1-C6 alkyl)(C3-C 14 carbocyclyl), C6-C 10 Aryl, 3-14 membered heterocyclic -(C1-C6 alkyl)-(3-14 membered heterocycloalkyl), where 1-4 members of the heterocycloalkyl are independently selected from N, O, and S; and -(C1-C6 alkyl)-(3-14 membered heterocycloalkyl), where 1-4 members of the heterocycloalkyl are independently selected from N, O, and S; and -(C1-C6 alkyl)-(3-14 membered heterocycloalkyl), where 1-4 members of the heteroaryl are independently selected from N, O, and S. are independently selected from the group consisting of In the formula, R A and R B Each alkyl, alkoxy, alkenyl, alkynyl, aryl, carbocyclyl, heterocycloalkyl, and heteroaryl moiety in 10 Aryl, 3- to 14-membered heterocycloalkyl, and -(C1-C6 alkyl)-(3- to 14-membered heterocycloalkyl), where 1-4 members of the ring are independently selected from N, O, and S, and 5- to 10-membered heteroaryl, where 1-4 members of the heteroaryl are independently selected from N, O, and S. and S), —NHC(O)(OC1-C6 alkyl), —NO2, —CN, oxo, —C(O)OH, —C(O)O(C1-C6 alkyl), —C1-C6 alkyl(C1-C6 alkoxy), -C(O)NH2, C1-C6 alkyl, -C(O)C1-C6 alkyl, -OC1-C6 alkyl, -Si(C1-C6 alkyl)3, -S(O) 0-2 -(C1-C6 alkyl), C6-C 10 Aryl, -(C1-C6 alkyl)(C6-C 10 a -(C-C alkyl), 3- to 14-membered heterocycloalkyl, and -(C-C alkyl)-(3- to 14-membered heterocycle), where 1-4 members of the heterocycle are independently selected from N, O, and S, and -O(C-C 14aryl), Therefore, In the formula, R A and R B Each alkyl, alkenyl, aryl, and heterocycloalkyl substituent is selected from the group consisting of hydroxy, —OC1-C6 alkyl, halo, —NH2, —(C1-C6 alkyl)NH2, —C(O)OH , CN, and oxo; R C and R D are each independently selected from H and C1-C6 alkyl, or a pharmaceutically acceptable salt thereof. (Item 2) During the ceremony, R 4 is H, C1-C 6- Alkyl (one or more halo, hydroxy, or 3-14 membered hetero heterocycloalkoxy (wherein 1 to 4 members of heterocycloalkoxy are independently selected from N, O, and S), —O(C1-C6-alkyl), —(C1-C6 alkyl)NR A R B , and -NR A R B (where R A and R B are independently selected from H and C1-C6 alkyl; R 5 is H, C1-C6 alkyl, C1-C6 alkoxy, and -NR C R D selected from the group consisting of , the compound according to item 1. (Item 3) R 4 and R 5 The compound according to item 1 or 2, wherein at least one of (Item 4) R 4 is H. (Item 5) R 5 is H. (Item 6) R 4 and R 5 Each of the following is H: (Item 7) R 2 optionally substituted C6-C 10 Aryl or optionally substituted 5- to 10-membered heteroaryl 7. The compound according to any one of items 1 to 6, wherein the compound is a benzoyl group. (Item 8) R 2 is arbitrarily substituted C 6- C 10 8. The compound according to item 7, wherein the compound is aryl. (Item 9) R 2 9. The compound according to item 8, wherein is optionally substituted phenyl. (Item 10) R 2 is an optionally substituted 5-10 membered heteroaryl, and one ring member is N. (Item 11) R 2 is an optionally substituted 5- or 6-membered heteroaryl. (Item 12) R 2 is an optionally substituted 6-membered heteroaryl. . (Item 13) R 2 13. The compound according to any one of items 10 to 12, wherein is optionally substituted pyridyl. (Item 14) R 3 is an optionally substituted 3- to 14-membered heterocycloalkyl or an optionally substituted 5- to 10-membered heteroaryl. (Item 15) R3 is selected from the group consisting of benzothiazolyl, benzisothiazolyl, benzoxazolyl, pyridinyl, pyridinonyl, pyridazinyl, benzimidazolyl, benzotriazolyl, indazolyl, quinoxalinyl, quinolinyl, quinazolinyl, imidazopyridinyl, pyrazolopyridinyl, triazolopyridinyl, cinnolinyl, isoxazolyl, pyrazolyl, benzofuranyl, dihydrobenzofuranyl, dihydrobenzodioxinyl, and tetrahydrobenzodioxinyl, each of which can be optionally substituted. (Item 16) R 3 optionally substituted C6-C 10 13. The compound according to any one of items 1 to 12, wherein the compound is aryl. Compound. (Item 17) R 3 17. The compound according to item 16, wherein is optionally substituted phenyl. (Item 18) R 2 is optionally substituted phenyl, and R 3 3-14 membered heterocyclic rings optionally substituted Any of items 1 to 6, wherein the aryl is chloroalkyl or optionally substituted 5- to 10-membered heteroaryl. The compound according to any one of claims 1 to 4. (Item 19) 19. The compound according to any one of items 1 to 18, wherein L is O or NR. (Item 20) R 1 is arbitrarily substituted C 1- C alkyl or optionally substituted C 3- Item 19. The compound according to item 19, which is a C6 carbocyclyl. (Item 21) R 1 is C1-C3 alkyl optionally substituted with 1 to 3 F. (Item 22) During the ceremony, L is O or NR and R is H; R1 is C1-C3 alkyl optionally substituted with 1 to 3 F, R 2 is an optionally substituted 3- to 14-membered heterocycloalkyl or an optionally substituted 5- to 10-membered heteroaryl (wherein one member of the heterocycloalkyl or heteroaryl is N) or an optionally substituted C-C 10 is aryl, R 3 is an optionally substituted 3- to 14-membered heterocycloalkyl or an optionally substituted 5- to 10-membered heteroaryl, wherein 1-3 members of the heterocycloalkyl or heteroaryl are independently selected from N, O, and S; and R 4 and R 5 and each of is H. (Item 23) 23. The compound according to item 22, wherein L is NR. (Item 24) The compound according to item 1 or a pharmaceutically acceptable salt thereof is selected from the following table: Acceptable salt. [Table 14-1] [Table 14-2] [Table 14-3] [Table 14-4] [Table 14-5] [Table 14-6] [Table 14-7] [Table 14-8] [Table 14-9] [Table 14-10] [Table 14-11] [Table 14-12] (Item 25) The compound according to item 1 or a pharmaceutically acceptable salt thereof is selected from the following table: Acceptable salt. [Table 15-1] [Table 15-2] [Table 15-3] [Table 15-4] (Item 26) A therapeutically effective compound according to any one of items 1 to 25 or a pharmaceutically acceptable salt thereof. and a pharmaceutically acceptable carrier. (Item 27) A method of treating cancer in a subject suffering from cancer, comprising administering to said subject a compound according to any one of items 1 to 25. A method comprising administering an effective amount of a MAT2A inhibitor compound described in any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof. (Item 28) 28. The method of item 27, wherein the cancer is an MTAP-deficient cancer. (Item 29) A method of treating cancer in a subject suffering from cancer, comprising administering to said subject a compound according to any one of items 1 to 25. or a pharmaceutically acceptable salt thereof. and (Item 30) 30. The method of item 29, wherein the cancer is an MTAP-deficient cancer. (Item 31) The cancers include mesothelioma, neuroblastoma, rectal cancer, colorectal cancer, familial adenomatous polyposis cancer and hereditary non-polyposis colorectal cancer, esophageal cancer, labial cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, gastric cancer, adenocarcinoma, medullary thyroid cancer, papillary thyroid cancer, kidney cancer, renal parenchymal cancer, ovarian cancer, cervical cancer, uterine cancer, endometrial cancer, choriocarcinoma, pancreatic cancer, prostate cancer, bladder cancer, testicular cancer, breast cancer, and urinary tract cancer. 30. The method according to any one of items 27 to 29, wherein the tumor is selected from the group consisting of carcinoma, melanoma, brain tumor, lymphoma, head and neck cancer, acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), hepatocellular carcinoma, gallbladder cancer, bronchial carcinoma, small cell lung cancer, non-small cell lung cancer, multiple myeloma, basal cell carcinoma, teratoma, retinoblastoma, choroidal melanoma, seminoma, rhabdomyosarcoma, osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma, Ewing's sarcoma, and plasmacytoma. (Item 32) 31. The method of item 29 or 30, wherein the cancer is selected from the group consisting of B-cell acute lymphocytic leukemia (B-ALL), mesothelioma, lymphoma, pancreatic cancer, lung cancer, gastric cancer, esophageal cancer, bladder cancer, brain tumor, head and neck cancer, melanoma and breast cancer. (Item 33) 33. The method of claim 32, wherein the cancer is a lung cancer selected from the group consisting of non-small cell lung cancer, small cell lung cancer, adenocarcinoma of the lung, and squamous cell carcinoma of the lung. (Item 34) 33. The method of claim 32, wherein the cancer is a brain tumor selected from the group consisting of glioma, glioblastoma, astrocytoma, meningioma, medulloblastoma, peripheral neuroectodermal tumor, and craniopharyngioma. (Item 35) 33. The method of claim 32, wherein the cancer is triple-negative breast cancer (TNBC). (Item 36) The cancer is mantle cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, Burkitt lymphoma, diffuse large B-cell lymphoma, and adult T-cell leukemia / lymphoma. 33. The method of claim 32, wherein the lymphoma is selected from the group consisting of: (Item 37) 26. A method for treating cancer in a subject suffering from cancer, comprising administering to said subject a compound according to any one of paragraphs 1 to 25. The compound or a pharmaceutically acceptable salt thereof. (Item 38) 38. The compound according to item 37, or a pharmaceutically acceptable salt thereof, wherein the cancer is an MTAP-deficient cancer. (Item 39) The cancers include mesothelioma, neuroblastoma, rectal cancer, colorectal cancer, familial adenomatous polyposis cancer and hereditary non-polyposis colorectal cancer, esophageal cancer, labial cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, gastric cancer, adenocarcinoma, medullary thyroid cancer, papillary thyroid cancer, kidney cancer, renal parenchymal cancer, ovarian cancer, cervical cancer, uterine cancer, endometrial cancer, choriocarcinoma, pancreatic cancer, prostate cancer, bladder cancer, testicular cancer, breast cancer, and urinary tract cancer. 39. The compound according to item 37 or 38, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of carcinoma, melanoma, brain tumor, lymphoma, head and neck cancer, acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), hepatocellular carcinoma, gallbladder cancer, bronchial carcinoma, small cell lung cancer, non-small cell lung cancer, multiple myeloma, basal cell carcinoma, teratoma, retinoblastoma, choroidal melanoma, seminoma, rhabdomyosarcoma, osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma, Ewing's sarcoma, and plasmacytoma. (Item 40) 39. The compound according to item 37 or 38, or a pharmaceutically acceptable salt thereof, wherein the cancer is selected from the group consisting of B-cell acute lymphocytic leukemia (B-ALL), mesothelioma, lymphoma, pancreatic cancer, lung cancer, gastric cancer, esophageal cancer, bladder cancer, brain tumor, head and neck cancer, melanoma and breast cancer. (Item 41) 41. The compound according to item 40, or a pharmaceutically acceptable salt thereof, wherein the cancer is a lung cancer selected from the group consisting of non-small cell lung cancer, small cell lung cancer, adenocarcinoma of the lung, and squamous cell carcinoma of the lung. (Item 42) 41. The compound according to item 40, or a pharmaceutically acceptable salt thereof, wherein the cancer is triple-negative breast cancer (TNBC). (Item 43) 41. The compound according to item 40, or a pharmaceutically acceptable salt thereof, wherein the cancer is a brain tumor selected from the group consisting of glioma, glioblastoma, astrocytoma, meningioma, medulloblastoma, peripheral neuroectodermal tumor, and craniopharyngioma. (Item 44) 40. The compound according to any one of items 37 to 39, or a pharmaceutically acceptable salt thereof, wherein the cancer is a lymphoma selected from the group consisting of mantle cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, Burkitt's lymphoma, diffuse large B-cell lymphoma (DLBCL), and adult T-cell leukemia / lymphoma. (Item 45) A compound according to any one of items 1 to 25 for the manufacture of a medicament for treating cancer. 2. Use of the compound according to claim 1, or a pharmaceutically acceptable salt thereof. (Item 46) 46. ​​The use according to item 45, or a pharmaceutically acceptable salt thereof, wherein the cancer is an MTAP-deficient cancer. (Item 47) The cancers include mesothelioma, neuroblastoma, rectal cancer, colorectal cancer, familial adenomatous polyposis cancer and hereditary non-polyposis colorectal cancer, esophageal cancer, labial cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, gastric cancer, adenocarcinoma, medullary thyroid cancer, papillary thyroid cancer, kidney cancer, renal parenchymal cancer, ovarian cancer, cervical cancer, uterine cancer, endometrial cancer, choriocarcinoma, pancreatic cancer, prostate cancer, bladder cancer, testicular cancer, breast cancer, and urinary tract cancer. 47. The use according to item 45 or 46, wherein the tumor is selected from the group consisting of carcinoma, melanoma, brain tumor, lymphoma, head and neck cancer, acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), hepatocellular carcinoma, gallbladder cancer, bronchial carcinoma, small cell lung cancer, non-small cell lung cancer, multiple myeloma, basal cell carcinoma, teratoma, retinoblastoma, choroidal melanoma, seminoma, rhabdomyosarcoma, osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma, Ewing's sarcoma and plasmacytoma, or a pharmaceutically acceptable salt thereof. (Item 48) 47. The use according to item 45 or 46, or a pharmaceutically acceptable salt thereof, wherein the cancer is selected from the group consisting of B-cell acute lymphocytic leukemia (B-ALL), mesothelioma, lymphoma, pancreatic cancer, lung cancer, gastric cancer, esophageal cancer, bladder cancer, brain tumor, head and neck cancer, melanoma and breast cancer. (Item 49) 49. The use according to item 48, or a pharmaceutically acceptable salt thereof, wherein the cancer is a lung cancer selected from the group consisting of non-small cell lung cancer, small cell lung cancer, adenocarcinoma of the lung, and squamous cell carcinoma of the lung. (Item 50) 49. The use according to item 48, or a pharmaceutically acceptable salt thereof, wherein the cancer is triple-negative breast cancer (TNBC). (Item 51) 49. The use according to item 48, or a pharmaceutically acceptable salt thereof, wherein the cancer is a brain tumor selected from the group consisting of glioma, glioblastoma, astrocytoma, meningioma, medulloblastoma, peripheral neuroectodermal tumor, and craniopharyngioma. (Item 52) 49. The use according to item 48, or a pharmaceutically acceptable salt thereof, wherein the cancer is a lymphoma selected from the group consisting of mantle cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, Burkitt lymphoma, diffuse large B-cell lymphoma (DLBCL), and adult T-cell leukemia / lymphoma. DETAILED DESCRIPTION OF THE INVENTION

[0028] The compounds described herein are MAT2A inhibitors. Accordingly, the present disclosure relates not only to such compounds conforming to general formula I, but also to their pharmaceutical compositions, tautomers, and isotopologues. The compounds and compositions are useful for treating cancer. Some cancers include various MTAP-deficient cancers, i.e., cancers in which the MTAP gene / protein is absent or deleted, or or cancers characterized by reduced function of the MTAP protein.

[0029] definition "Alkyl" refers to straight and branched chain hydrocarbyl groups of 1 to about 20 carbon atoms. For example, alkyl can have 1 to 10 carbon atoms or 1 to 6 carbon atoms. Exemplary alkyl includes straight chain alkyl groups such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, and the like, and also includes, for example, -CH(CH3)2, -CH(CH3)(CH2CH3), -CH(CH2CH3)2, -C(CH3)3, -C(CH2CH3)3, -CH2CH(CH3)2, -CH2CH(CH3)(CH2CH3), -CH2CH(CH2CH3)2, -CH2C(CH3)3, -CH2C(CH2CH3)3, -CH(CH3)CH(CH3)(CH2CH3), -CH2CH2CH(CH3)2, -CH2CH2CH(CH3)(CH2CH3), -CH2CH2CH(CH2CH3)2, -CH2CH2CH(CH3)(CH2CH3), -CH2CH2CH(CH2CH3)2, -CH2CH2C(CH3)3, -CH2CH2C(CH2CH3)3, Includes, but is not limited to, branched chain isomers of straight chain alkyl groups such as -CH(CH3)CH2CH(CH3)2, -CH(CH3)CH(CH3)CH(CH3)2, Thus, alkyl groups include primary alkyl groups, secondary alkyl groups, and tertiary alkyl groups. Alkyl groups can be unsubstituted or optionally substituted with one or more substituents described below.

[0030] The phrase "substituted alkyl" refers to an alkyl substituted at one or more positions, e.g., 1, 2, 3, 4, 5, or even 6 positions, and includes any of the substitutions described herein. The substituent may be attached to any available atom which results in a stable compound. "Optionally substituted alkyl" refers to alkyl or substituted alkyl.

[0031] The terms "halogen," "halide," and "halo" each refer to -F, -Cl, -Br, or -I.

[0032] The term "alkenyl" refers to 1 to 3, 1 to 2, or at least one carbon-carbon dialkyl group. A straight or branched chain hydrocarbyl group containing 2 to about 20 carbon atoms and having a double bond. An alkenyl group can be unsubstituted or optionally substituted with one or more substituents described herein below.

[0033] "Substituted alkenyl" refers to an alkenyl substituted at one or more positions, for example, 1, 2, 3, 4, 5, or even 6 positions, where the substitutions described herein allow the substituent to be attached to any available atom resulting in a stable compound. "Optionally substituted alkenyl" refers to an alkenyl or substituted alkenyl.

[0034] "Alkyne or "alkynyl" refers to a straight- or branched-chain unsaturated hydrocarbon having the indicated number of carbon atoms and at least one triple bond. Examples of alkynyl groups include, but are not limited to, acetylene, propyne, 1-butyne, 2-butyne, 1-pentyne, 2-pentyne, 1-hexyne, 2-hexyne, 3-hexyne, 1-heptyne, 2-heptyne, 3-heptyne, 1-octyne, 2-octyne, 3-octyne, and 4-octyne. Alkynyl groups can be unsubstituted or optionally substituted with one or more substituents described herein below.

[0035] "Substituted alkynyl" refers to alkynyl groups that are substituted, for example, at one, two, three, four, five, or even six positions. "Optionally substituted alkynyl" refers to an alkynyl substituted at one or more positions, where the substitutions described herein allow the substituent to be attached to any available atom which results in a stable compound. "Optionally substituted alkynyl" refers to an alkynyl or substituted alkynyl.

[0036] The term "alkoxy" refers to an --O-alkyl group having the indicated number of carbon atoms. For example, the (C1-C6)alkoxy group includes -O-methyl, -O-ethyl, -O-propyl, -O-iso Examples include propyl, -O-butyl, -O-sec-butyl, -O-tert-butyl, -O-pentyl, -O-isopentyl, -O-neopentyl, -O-hexyl, -O-isohexyl, and -O-neohexyl.

[0037] The term "carbocyclyl" refers to a monocyclic, bicyclic, tricyclic, or polycyclic refers to a 3- to 14-membered ring system, which may be saturated or unsaturated, e.g., "cycloalkyl" The term "cycloalkenyl" specifically refers to cyclic alkenyl, e.g., C3-C6 cycloalkenyl. Carbocyclyl may be bonded via any atom. For example, carbocyclyl also contemplates fused rings, e.g., a carbocyclyl fused to an aryl or heteroaryl ring as defined herein. Representative examples of carbocyclyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, phenyl, naphthyl, anthracyl, benzofuranyl, and benzothiephenyl. A carbocyclyl group can be unsubstituted or optionally substituted with one or more substituents described herein below.

[0038] "Substituted carbocyclyl" refers to a 1-substituted carbocyclyl, e.g., 1, 2, 3, 4, 5, or even 6-position 1 "Optionally substituted carbocyclyl" refers to a carbocyclyl substituted at one or more positions, where the substitution is as described herein, and the substituent may be attached to any available atom to result in a stable compound. "Optionally substituted carbocyclyl" refers to a carbocyclyl or substituted carbocyclyl.

[0039] When used alone or as part of another term, "aryl" means any group having the specified number of carbon atoms or, if no number is specified, any group, e.g., C-C 14"aryl" refers to a fused or unfused carbocyclic aromatic group containing up to 14 carbon atoms, such as aryl. Particular aryl groups are phenyl, naphthyl, biphenyl, phenanthrenyl, naphthacenyl, and the like (see, for example, Lang's Handbook of Chemistry (Dean, JA, ed.), 13th Edition, Table 7-2

[1985] ). Particular aryl is phenyl. "Aryl" also includes aromatic ring systems, as defined herein, optionally fused with a carbocyclyl ring. Aryl groups can be unsubstituted or optionally substituted with one or more substituents, as described herein below.

[0040] "Substituted aryl" is an aryl independently substituted with one or more substituents attached at any available atom to result in a stable compound, the substituents being as described herein. "Optionally substituted aryl" refers to an aryl or substituted aryl.

[0041] The term "heteroatom" refers to N, O, and S. Compounds containing N or S atoms are , which may optionally be oxidized to the corresponding N-oxide, sulfoxide, or sulfone compound.

[0042] "Heteroaryl," alone or in combination with other moieties described herein, refers to any heteroaryl group independently selected from the group consisting of O, S, and N, e.g., 1 to 4, 1 to 3, or 1 to 2. 5 or 6 rings containing one or more heteroatoms, such as 5 or 6 heteroatoms Heteroaryl refers to a monocyclic aromatic ring structure containing 5 to 10 atoms, or a bicyclic aromatic group having 8 to 10 atoms. Heteroaryl is also intended to include oxidized S or N, such as sulfinyl, sulfonyl, and N-oxides of tertiary ring nitrogens. A carbon or heteroatom is the point of attachment of the heteroaryl ring structure such that a stable compound is produced. Examples of heteroaryl groups include pyridinyl, pyridazinyl, pyrazinyl, quinoxalyl, indolizinyl, benzo[b]thienyl, quinazolinyl, purinyl, indolyl, and quinazolinyl. Heteroaryl groups include, but are not limited to, aryl, pyrimidinyl, pyrrolyl, pyrazolyl, oxazolyl, thiazolyl, thienyl, isoxazolyl, oxathiadiazolyl, isothiazolyl, tetrazolyl, imidazolyl, triazolyl, furanyl, benzofuryl, and indolyl. Heteroaryl groups can be unsubstituted or optionally substituted with one or more substituents described herein below.

[0043] "Substituted heteroaryl" refers to a heteroaryl group, unless otherwise specified, that has one or more, e.g., 1, 2, 3, 4, or 5, substituted heteroaryl groups attached to any available atom to produce a stable compound. or heteroaryl independently substituted with one substituent that is also a substituent, which is also a substituent for three, wherein the substituents are as described herein. "Optionally substituted heteroaryl" refers to heteroaryl or substituted heteroaryl.

[0044] "Heterocycloalkyl" refers to a saturated alkyl group having, for example, 3 to 6 atoms, and 3 to 14 atoms. Heterocycloalkoxy means a saturated or unsaturated non-aromatic monocyclic, bicyclic, tricyclic or polycyclic ring system in which one to four carbon atoms in the ring are replaced by an O, S or N heteroatom. The heterocycloalkyl group is optionally fused to a 5- to 6-membered aryl or heteroaryl ring and includes oxidized S or N, such as sulfinyl, sulfonyl, and N-oxides of tertiary ring nitrogens. The point of attachment of the heterocycloalkyl ring is at a carbon or heteroatom such that a stable ring is maintained. Examples of heterocycloalkyl groups include, but are not limited to, morpholino, tetrahydrofuranyl, dihydropyridinyl, piperidinyl, pyrrolidinyl, piperazinyl, dihydrobenzofuryl, and dihydroindolyl. Heterocycloalkyl groups can be unsubstituted or optionally substituted with one or more substituents described herein below.

[0045] An "optionally substituted heterocycloalkyl" is a heterocycloalkyl group substituted with 1 to 3 substituents, e.g., 1, 2, or 3 substituents, attached at any available atom to produce a stable compound. "Hydroxy" refers to an substituted heterocycloalkyl, wherein the substituents are as described herein.

[0046] "Heterocycloalkoxy" refers to a monocyclic, bicyclic, tricyclic, or polycyclic ring system having the indicated number of member atoms, in which one to four of the carbon atoms in the ring are heteroatoms of O, S, or N. refers to an -O-heterocycloalkyl group substituted with aryl.

[0047] "Optionally substituted heterocycloalkoxy" refers to a heterocycloalkoxy group having 1 to 3 substituents attached at any available atom to produce a stable compound, e.g., 1, 2, or 3 substituents. refers to a substituted heterocycloalkoxy, wherein the substituents are as described herein.

[0048] The terms "nitrile" or "cyano" can be used interchangeably and refer to a -CN group attached to carbon atoms of heteroaryl, aryl, and heterocycloalkyl rings.

[0049] The term "oxo" refers to an =0 atom bonded to a saturated or unsaturated moiety. The =0 atom can be bonded to a carbon, sulfur, or nitrogen atom that is part of a cyclic or acyclic moiety.

[0050] "Hydroxyl" or "hydroxy" refers to the group --OH.

[0051] The substituent -COH may be, for example, [ka] and the like, where R is R as defined herein. A For example, The Practice of Medicinal Chemistry Practice) (Academic Press: New York, 1996), p. 203.

[0052] The compounds described herein may include configurational isomers, geometric isomers, and isomers such as cis- or cis-isomeric isomers. It can exist in various isomeric forms, including conformational isomers, including trans- and di-isomers. A compound may exist in one or more tautomers, including both a single tautomer and a mixture of tautomers. The term "isomer" is intended to encompass all isomers of the compounds of the present disclosure, including tautomers of the compound. The compounds of the present disclosure may also exist in open-chain or cyclized forms. In some cases, one or more cyclized forms may result from the loss of water. The specific composition of the open-chain and cyclized forms may depend on how the compound is isolated, stored, or administered. For example, a compound may exist primarily in an open-chain form under acidic conditions, but may be in a cyclized form under neutral conditions. All forms are included in the present disclosure.

[0053] Some compounds described herein have asymmetric centers and can exist in various enantiomers and diastereoisomers.The compounds of the present disclosure can be in the form of optical isomers or diastereomers.Therefore, the present disclosure encompasses the compounds described herein and their use in the form of their optical isomers, diastereoisomers and mixtures thereof, including racemic mixtures.The optical isomers of the compounds of the present disclosure can be obtained by known techniques such as asymmetric synthesis, chiral chromatography, simulated moving bed technology, or by chemical separation of stereoisomers using optically active resolving agents.

[0054] Unless otherwise indicated, the term "stereoisomer" means one stereoisomer of a compound that is substantially free of other stereoisomers of that compound. Thus, a stereoisomerically pure compound having one chiral center will be substantially free of the opposite enantiomer of that compound. A stereoisomerically pure compound having two chiral centers will be substantially free of other diastereomers of that compound. A typical stereoisomerically pure compound will contain greater than about 80% by weight of one stereoisomer of the compound and less than about 80% by weight of the other stereoisomer of the compound. For example, the compound may contain more than about 90% by weight of one stereoisomer of the compound and less than about 10% by weight of the other stereoisomer of the compound, or more than about 95% by weight of one stereoisomer of the compound. % by weight of the other stereoisomer of the compound and less than about 5% by weight of one stereoisomer of the compound greater than about 97% by weight of one stereoisomer of the compound and less than about 3% by weight of the other stereoisomer of the compound, or A compound containing greater than about 99% by weight of one stereoisomer and less than about 1% by weight of the other stereoisomer of the compound. The stereoisomers described above can be viewed as compositions comprising the two stereoisomers present in their respective weight percentages as set forth herein.

[0055] As used herein, the term "isotopologue" refers to an isotopically enriched compound. As used herein, unless otherwise indicated, the term "isotopically enriched" refers to an atom having an isotopic composition other than that atom's naturally abundant isotopic composition. "Isotopically enriched" may also refer to a compound containing at least one atom having an isotopic composition other than that atom's natural isotopic composition. In an isotopologue, "isotopic enrichment" refers to the proportion of a specific isotope of a given atom incorporated into a molecule, instead of that atom's natural isotopic composition. For example, 1% deuterium enrichment at a given position means that 1% of the molecules in a given sample contain deuterium at that particular position. Because the naturally occurring distribution of deuterium is approximately 0.0156%, a compound synthesized using non-enriched starting materials The deuterium enrichment at any position in is approximately 0.0156%.

[0056] Thus, as used herein, unless otherwise indicated, the term "isotopic enrichment factor" refers to the ratio between the isotopic composition of a specified isotope and the natural isotopic composition.

[0057] With respect to the compounds provided herein, certain atom positions may be designated as deuterium or "D" or is "H 2 When a position is designated as having "a" or "an" atom, it is understood that the abundance of deuterium at that position is substantially greater than the natural abundance of deuterium, which is about 0.015%. A position designated as having deuterium will typically have at least 1000 (15% deuterium incorporation), and at least 1000 (15% deuterium incorporation), in certain embodiments. and a minimum isotopic enrichment factor for each specified deuterium atom of at least 2000 (30% deuterium incorporation), at least 3000 (45% deuterium incorporation), at least 3500 (52.5% deuterium incorporation), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation). The isotopic enrichment and isotopic enrichment factor of the compounds provided herein can be determined using conventional analytical methods known to those skilled in the art, including mass spectrometry and nuclear magnetic resonance spectroscopy.

[0058] In the event of a discrepancy between a depicted structure and the name given to that structure, the depicted structure shall be considered. Furthermore, if the stereochemistry of a structure or portion of a structure is not shown, for example, by bold or dashed lines, the structure or portion of the structure shall be interpreted as encompassing all stereoisomers thereof. However, in some cases, when multiple chiral centers are present, the structure and name may be represented as a single enantiomer to help explain the relevant stereochemistry. Those skilled in the art of organic synthesis will know whether a compound is prepared as a single enantiomer from the method used to prepare it.

[0059] As used herein, and unless specifically stated to the contrary, the term "compound" is inclusive in that it encompasses the compound or its pharmaceutically acceptable salts, stereoisomers, isotopologues, and / or tautomers. For example, compounds of general formula I or II may be pharmaceutically acceptable salts of isotopologues of the compounds. include.

[0060] As used herein, "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable salt of an organic or inorganic acid or base of the compound of the present disclosure. Representative pharmaceutically acceptable salts include, for example, alkali metal salts, alkaline earth salts, ammonium salts, such as acetate, amsonate (4,4-diaminostilbene-2,2-disulfonate), benzenesulfonate, Benzonate, bicarbonate, bisulfate, tartrate, borate, bromide, Butyrate, calcium, calcium edetate, camsylate, carbonate, chloride, citrate, clavulariate, dihydrochloride, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate , glutamate, glycolyl arsanilate, hexafluorophosphate, hexylresorcinate, hydrabamine ), hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isothionate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methyl bromide, methyl nitrate, methyl sulfate, mucate, napsylate, nitrate, N-methylglucamine ammonium salt, 3-hydroxy-2-naphthoate, oleate, oxalate Salt, palmitate, pamoate (1,1-methene-bis-2-hydroxy-3-naphthohydrochloride, einbonate, pantothenate, phosphate / diphosphate, picrate, polygalacturonate, propionate, p-toluenesulfonate, salicylate, stearate, diacetate, succinate, sulfate, sulfosalicylate Pharmaceutically acceptable salts include water-soluble salts and water-insoluble salts such as suramate, tannate, tartrate, teoclate, tosylate, triethyl iodide, and valerate. Pharmaceutically acceptable salts can have one or more charged atoms in their structure. In this case, the pharmaceutically acceptable salt can have multiple counter ions. Thus, the pharmaceutically acceptable salt can have one or more charged atoms and / or one or more counter ions.

[0061] The terms "treat," "treating," and "treatment" refer to the amelioration or eradication of a disease or symptoms associated with a disease. In certain embodiments, these terms refer to minimizing the spread or worsening of a disease that results from the administration of one or more prophylactic or therapeutic agents to a patient suffering from such a disease.

[0062] The terms "prevent," "preventing," and "prevention" refer to the prevention of the onset, recurrence, or spread of disease in a patient resulting from the administration of a prophylactic or therapeutic agent.

[0063] The term "effective amount" refers to an amount of a compound or other active ingredient of the present disclosure sufficient to provide a therapeutic or prophylactic benefit in the treatment or prevention of a disease, or to delay or minimize symptoms associated with a disease. Furthermore, with respect to a compound of the present disclosure, a therapeutically effective amount means an amount of a therapeutic agent alone, or in combination with other therapies, that provides a therapeutic benefit in the treatment or prevention of a disease. When used in connection with a compound of the present disclosure, the term can encompass an amount that improves overall therapy, reduces or avoids the symptoms or causes of a disease, or enhances or synergizes with another therapeutic agent.

[0064] A "patient" or "subject" includes animals such as humans, cows, horses, sheep, lambs, pigs, chickens, turkeys, quail, cats, dogs, mice, rats, rabbits, or guinea pigs. According to some embodiments, the animals are mammals, such as non-primates and primates (e.g., monkeys and humans). In one embodiment, the patient is a human, such as a human infant, child, adolescent, or adult.

[0065] "Inhibitor" refers to a compound that prevents or reduces the amount of SAM synthesis. In certain embodiments, the inhibitor binds to MAT2A. In one embodiment, the inhibitor inhibits the function of MAT2A.

[0066] compound As generally described above, the present disclosure provides compounds, as well as pharmaceutically acceptable salts, tautomers, and / or isotopologues thereof, which compounds conform to the general formula I: [ka]

[0067] In general formula I, L is O, S, NR, or a bond. R is H or C1-C6 alkyl.

[0068] R 1 is selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C3-C6 carbocyclyl, -(C1-C6 alkyl)(C3-C6 carbocyclyl), and -(C1-C6 alkyl)(C3-C6 cycloalkenyl); wherein R 1 Any alkyl in R is linear or branched. 1 is optionally substituted by 1 to 6 halo or 1 to 6 deuterium.

[0069] Alternatively, in one embodiment, when L is NR, R and R in combination with L 1 is one One or more R Aand represents a 3- to 6-membered heterocycloalkyl optionally substituted with , wherein 1-4 ring members are independently selected from N, O, and S.

[0070] R 2 and R 3 is C2-C6 alkynyl, C6-C 10 Aryl, C3-C6 carbocyclyl, 5-10 membered Heteroaryl (wherein 1 to 4 members of the heteroaryl are independently selected from N, O, and S) and 3- to 14-membered heterocycloalkyl (wherein 1 to 4 of the heterocycloalkyl R 2 Oh BiR 3 is R A , OR A , halo, -N=NR A , N.R. A R B , -(C1-C6 alkyl)NR A R B , -C(O)OR A , -C(O)NR A R B , -OC(O)R A , —SI(C1-C6 alkyl)3, and —CN; or It is independently and optionally substituted by multiple substituents.

[0071] R 4 is H, C1-C6 alkyl (optionally one or more halo, hydroxy, or 3-14 membered heterocycloalkoxy (wherein 1-4 members of the heterocycloalkoxy are independently selected from N, O, and S)), —O(C1-C 6- alkyl) (optionally substituted with one or more halo -OH, halo, -CN, -(C1-C6 alkyl)NR A R B , and -NR A R B Selected from the group consisting of can be.

[0072] R 5 is H, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, Halo, -CN, and -NR C R D is selected from the group consisting of:

[0073] R A and R B is H, -CN, -hydroxy, oxo, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, -NH2, -S(O) 0-2 -(C1-C6 alkyl), -S(O) 0-2 -(C6-C 10 aryl), -C(O)(C1-C6 alkyl), -C(O)(C3-C 14 carbocyclyl), -C3-C 14 Carbosic -(C1-C6 alkyl)(C3-C 14 carbocyclyl), C6-C 10 Aryl, 3-14 membered heterocyclic -(C1-C6 alkyl)-(3-14 membered heterocycloalkyl), where 1-4 members of the heterocycloalkyl are independently selected from N, O, and S; and -(C1-C6 alkyl)-(3-14 membered heterocycloalkyl), where 1-4 members of the heterocycloalkyl are independently selected from N, O, and S; and -(C1-C6 alkyl)-(3-14 membered heterocycloalkyl), where 1-4 members of the heteroaryl are independently selected from N, O, and S. are independently selected from the group consisting of:

[0074] R A and R B Each alkyl, alkoxy, alkenyl, alkynyl, aryl, carbocyclyl, heterocycloalkyl, and heteroaryl moiety in 10Aryl, 3- to 14-membered heterocycloalkyl, and -(C1-C6 alkyl)-(3- to 14-membered heterocycloalkyl), where 1-4 ring members are independently selected from N, O, and S. and 5-10 membered heteroaryl (wherein 1-4 members of the heteroaryl are independently selected from N, O, and S), —NHC(O)(O—C alkyl), —NO, —CN, oxo, —C(O)OH, —C(O)O(C alkyl), —C-C alkyl (C alkoxy). Si), -C(O)NH2, C1-C6 alkyl, -C(O)C1-C6 alkyl, -OC1-C6 alkyl, -Si(C1-C6 alkyl)3, -S(O) 0-2 -(C1-C6 alkyl), C6-C 10 Aryl, -(C1-C6 alkyl)(C6-C 10 Ally -(C-C alkyl), 3- to 14-membered heterocycloalkyl, and -(C-C alkyl)-(3- to 14-membered heterocycle), where 1-4 members of the heterocycle are independently selected from N, O, and S, and -O(C-C 14 aryl). A and R B of Each alkyl, alkenyl, aryl, and heterocycloalkyl substituent is optionally substituted with one or more substituents selected from the group consisting of hydroxy, —O—C alkyl, halo, —NH, —(C—C alkyl)NH, —C(O)OH, CN, and oxo.

[0075] R C and R D are each independently selected from H and C1-C6 alkyl.

[0076] In some embodiments, R 4 is H, C1-C6 alkyl (one or more halo, hydroxy or 3-14 membered heterocycloalkoxy (wherein 1-4 membered heterocycloalkoxy is independently selected from N, O, and S), —O(C1-C6 alkyl), —(C1-C6-alkyl)NR A R B , and -NR A R B (where R A and R B are independently selected from H and C1-C6 alkyl; R 5 is H, C1-C6 alkyl, C1-C6 alkoxy -xy, and -NR C R D is selected from the group consisting of:

[0077] In other embodiments, R 4 and R 5 At least one of R is H. For example, 4 is H or R 5 is H. Alternatively, R 4 and R 5 Each of is H.

[0078] Optionally, in combination with any other embodiment described herein, various embodiments provide compounds of general formula I, wherein R 2 is an arbitrarily substituted C 6- C 10 Aryl or any In one embodiment, R 2 is arbitrarily replaced Optionally substituted C6-C 10 In another embodiment, R 2 is an optionally substituted 5-10 membered heteroaryl, where one ring member is N. Examples For example, R 2 is an optionally substituted 5- or 6-membered heteroaryl, or an optionally substituted 6-membered heteroaryl, an example of which is optionally substituted pyridyl.

[0079] In some embodiments, R 3 R is an optionally substituted 3- to 14-membered heterocycloalkyl or an optionally substituted 5- to 10-membered heteroaryl. 3 Non-limiting examples of are selected from the group consisting of benzothiazolyl, benzisothiazolyl, benzoxazolyl, pyridinyl, pyridinonyl, pyridazinyl, benzimidazolyl, benzotriazolyl, indazolyl, quinoxalinyl, quinolinyl, quinazolinyl, imidazopyridinyl, pyrazolopyridinyl, triazolopyridinyl, cinnolinyl, isoxazolyl, pyrazolyl, benzofuranyl, dihydrobenzofuranyl, dihydrobenzodioxinyl, and tetrahydrobenzodioxinyl, each of which can be optionally substituted.

[0080] In other embodiments, R 3 is an arbitrarily converted C6-C 10 An example is optionally substituted phenyl.

[0081] This disclosure is based on the R 2 is optionally substituted phenyl, and R 3 Optionally substituted 3- to 14-membered heterocyclic groups Some are heterocycloalkyl or optionally substituted 5-10 membered heteroaryl. The present invention provides compounds of the general formula I:

[0082] In various embodiments, the present disclosure provides compounds of general formula I, where L is O or NR. Furthermore, according to an additional embodiment, R 1 is an arbitrarily substituted C 1- C6 alkyl, or any C replaced by 3- C6 carbocyclyl. 1 An example of is C1-C3 alkyl optionally substituted with 1 to 3 F.

[0083] A subset of compounds of general formula I according to one embodiment is those in which L is O or NR and R is H. R 1 C optionally substituted with 1 to 3 F 1- C3 alkyl, and R 2 is an optionally substituted 3- to 14-membered heterocycloalkyl or an optionally substituted 5- to 10-membered heteroaryl (wherein wherein one member of the heterocycloalkyl or heteroaryl is N) or optionally substituted TaC 6- C 10 aryl, and R 3 is an optionally substituted 3- to 14-membered heterocycloalkyl or is an optionally substituted 5- to 10-membered heteroaryl, heterocycloalkyl or hetero 1 to 3 members of the aryl are independently selected from N, O, and S; R 4 and R 5 wherein each is H. In some embodiments, L is NR.

[0084] In various embodiments, the present disclosure provides compounds of general formula I, as well as pharmaceutically acceptable salts, tautomers, and / or isotopologues thereof, as set forth in Table 1 below. To provide a concrete example:

[0085] Table 1 [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Table 1-10] [Table 1-11]

[0086] In various other embodiments, the present disclosure provides compounds of general formula I, as set forth in Table 2 below, and their pharmaceutically acceptable salts, tautomers, and / or isotopologues. Further specific examples are provided:

[0087] Table 2 [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4]

[0088] Pharmaceutical Composition The present disclosure also provides a therapeutically effective amount of one or more compounds according to general formula I, or a pharmaceutical Pharmaceutically acceptable salts, stereoisomers, tautomers, and / or isotopologues may be used in combination with other compounds. Also provided are pharmaceutical compositions comprising the compound of formula (I) in admixture with a pharmaceutically acceptable carrier, in some embodiments, the composition further comprises one or more additional therapeutic agents, pharmaceutically acceptable excipients, diluents, adjuvants, stabilizers, emulsifiers, preservatives, colorants, buffers, flavoring agents, in accordance with accepted practice in pharmaceutical compounding.

[0089] In one embodiment, the pharmaceutical composition comprises a compound selected from those shown in Table 1, or and a pharmaceutically acceptable salt, stereoisomer, tautomer, and / or isotopologue of and a pharmaceutically acceptable carrier.

[0090] The pharmaceutical compositions of the present disclosure are formulated, dosed, and administered in a manner consistent with good medical practice. Factors to consider in this context include the particular disorder being treated, the particular subject being treated, the clinical condition of the subject, the cause of the disorder, the site of delivery of the agent, the method of administration, the administration schedule, and other factors known to medical professionals.

[0091] The compound to be administered (or its pharmaceutically acceptable salts, stereoisomers, tautomers, and A "therapeutically effective amount" of a compound (and / or isotopologues) will be governed by such considerations and may be an amount sufficient to exert a cytotoxic effect on the cancer, inhibit MAT2A activity, or both. Such an amount is likely to be less than an amount that is toxic to normal cells, or to the subject as a whole. Generally, the compound of the present disclosure (or its pharmaceutically acceptable salts) administered The initial therapeutically effective amount of the compound (or a tolerable salt, stereoisomer, or tautomer) is 100 mg / kg of the compound per patient per day. The dose ranges from about 0.01 to about 200 mg / kg or from about 0.1 to about 20 mg / kg of body weight, with a typical initial range being about 0.3 to about 15 mg / kg / day. Oral unit dosage forms, such as tablets and capsules, contain from about 1 mg to about 1000 mg of a compound of the present disclosure (or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof). In another embodiment, such dosage forms contain from about 50 mg to about 500 mg of a compound of the present disclosure (or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof). In yet another embodiment, such dosage forms contain from about 25 mg to about 200 mg of a compound of the present disclosure (or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof). In another embodiment, such dosage forms contain from about 10 mg to about 100 mg of a compound of the present disclosure (or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof). In some embodiments, such dosage forms contain from about 5 mg to about 50 mg of a compound of the present disclosure (or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof).

[0092] The compositions may be administered orally, topically, parenterally, by inhalation or aerosol or rectally in dosage unit formulations. As used herein, the term parenteral includes subcutaneous injections, intravenous, intramuscular, intrasternal injection or infusion techniques.

[0093] Suitable oral compositions according to the present disclosure include, but are not limited to, tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, syrups, or elixirs.

[0094] Pharmaceutical compositions suitable for single unit dosages comprising a compound of the present disclosure or a pharmaceutically acceptable stereoisomer, salt, or tautomer thereof and a pharmaceutically acceptable carrier are encompassed within the scope of the present disclosure.

[0095] Compositions suitable for oral use may be prepared according to any method known in the art for the manufacture of pharmaceutical compositions. For example, a liquid formulation of the compound may be prepared to provide a pharmaceutically palatable MAT2A inhibitor. In order to provide a palatable preparation, the preparation may contain one or more agents selected from the group consisting of sweetening agents, flavoring agents, coloring agents and preservatives.

[0096] For tablet compositions, the compound of the present disclosure mixed with non-toxic pharmaceutically acceptable excipients is used to manufacture tablets.Examples of such excipients include but are not limited to inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate, granulating and disintegrating agents such as corn starch or alginic acid, or binders such as starch, gelatin or acacia, and lubricants such as magnesium stearate, stearic acid or talc.Tablets can be uncoated or can be coated with known coating technology to delay disintegration and absorption in the gastrointestinal tract, thereby providing sustained therapeutic effects over a desired time period.For example, time-delay materials such as glyceryl monostearate or glyceryl distearate can be used.

[0097] Formulations for oral use may also be presented as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate, or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with an aqueous medium or an oil medium, such as peanut oil, liquid paraffin, or olive oil.

[0098] For aqueous suspensions, the compounds of the present disclosure are mixed with suitable excipients to maintain a stable suspension. Examples of such excipients include, but are not limited to, sodium carboxymethylcellulose, methylcellulose, hydropropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth, and gum acacia.

[0099] Oral suspensions may also contain dispersing or wetting agents such as naturally occurring phosphatides, for example, lectins, or condensation products of alkylene oxides with fatty acids, for example, polyoxyethylene stearate, or condensation products of ethylene oxide with long-chain aliphatic alcohols, for example, heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitols, for example, polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example, polyethylene sorbitan monooleate. Aqueous suspensions may also contain one or more preservatives, for example, ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, for example, sucrose or saccharin.

[0100] Oily suspensions may be formulated by suspending the compounds of the present disclosure in a vegetable oil, for example arachis oil, olive oil, sesame oil, or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions may contain a thickening agent, for example beeswax, hard paraffin, or cetyl alcohol.

[0101] Sweetening agents such as those set forth above, and flavoring agents may be added to provide a palatable oral preparation.These compositions may be preserved by the addition of an antioxidant such as ascorbic acid.

[0102] Dispersible powders and granules suitable for preparation of an aqueous suspension by adding water provide the compound of the present disclosure in admixture with a dispersing or wetting agent, a suspending agent, and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above. Additional excipients, such as sweeteners, flavorings, and coloring agents, may also be present.

[0103] The pharmaceutical composition of the present disclosure may also be in the form of an oil-in-water emulsion. The oil phase may be a vegetable oil, such as olive oil or peanut oil, or a mineral oil, such as liquid paraffin or a mixture thereof. Suitable emulsifiers may be natural gums, such as gum acacia or gum tragacanth, natural phosphatides, such as soybean, lecithin, and esters or partial esters derived from fatty acids, such as hexitols, anhydrides, such as sorbitan monooleate, and condensation products of the partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsifier may also contain sweeteners and flavoring agents.

[0104] Syrups and elixirs may be formulated with sweetening agents, such as glycerin, propylene glycol, sorbitol, or sucrose. These formulations may also contain demulcents, preservatives, and flavoring and coloring agents. The pharmaceutical compositions may be in the form of sterile injectable aqueous or oily suspensions. These suspensions may be formulated according to known techniques using suitable dispersing or wetting agents and suspending agents, as described above. Sterile injectable preparations may also be sterile injectable solutions or suspensions in non-toxic parenterally acceptable diluents or solvents, such as 1,3-butanediol solutions. Acceptable vehicles and solvents that may be employed include water, Ringer's solution, and isotonic sodium chloride solution. Additionally, sterile, fixed oils are conventionally employed as solvents or suspending media. For this purpose, any non-irritating fixed oil, including synthetic mono- or diglycerides, may be employed. Additionally, fatty acids, such as oleic acid, are used in the preparation of injectables.

[0105] The compounds of general formula I may also be administered in the form of suppositories for rectal administration of the drug. The composition can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at ambient temperature but liquid at rectal temperature and will therefore melt in the rectum to release the drug. Such materials are cocoa butter and polyethylene glycol.

[0106] The composition for parenteral administration is administered in sterile medium.Depending on the vehicle used and the concentration of drug in the preparation, parenteral preparation can be either suspension or solution containing dissolved drug.For example, adjuvants such as local anesthetic, preservative and buffer can also be added to parenteral composition.

[0107] How to use The MAT2A enzyme converts methionine and ATP into S-adenosylmethionine (SAM) in cells. Therefore, in another embodiment of the present disclosure, a method for inhibiting SAM synthesis in cells is provided. The method comprises the step of: reacting a compound of general formula I or a pharmaceutically acceptable salt, stereoisomer, tautomer, or the method comprising introducing into a cell an effective amount of the isomer and / or isotopologue. In some embodiments, the method further comprises administering to a subject a cell comprising administering to the subject an inhibitor of MAT2A, e.g., a competitive assay for binding to MAT2A or a competitive assay for inhibiting SAM production. The compounds of general formula I are used to identify other compounds that are Binding to MAT2A or inhibition of SAM production by a test compound can be measured with or without the presence of an unlabeled compound of the present disclosure.

[0108] The present disclosure also provides a method of treating cancer in a subject, comprising administering to the subject an effective amount of a MAT2A inhibitor compound described herein. In this embodiment, the MAT2A inhibitor is a compound of general formula I or a pharmaceutically acceptable salt thereof. In one embodiment, optionally in combination with any other embodiment, the subject is a mammal, such as a human.

[0109] In some embodiments, the cancer is an MTAP-deficient cancer. In some embodiments, the cancer is mesothelioma, neuroblastoma, intestinal cancer such as rectal cancer, colorectal cancer, familial adenomatous polyposis cancer and hereditary non-polyposis colorectal cancer, esophageal cancer, labial cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, gastric cancer, adenocarcinoma, medullary thyroid cancer, papillary thyroid cancer, kidney cancer, renal parenchymal cancer, ovarian cancer, cervical cancer, uterine cancer, endometrial cancer, choriocarcinoma, pancreatic cancer, prostate cancer, bladder cancer, testicular cancer, breast cancer, urinary tract cancer, carcinoma), melanoma, brain tumor, head and neck cancer, lymphoma, acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), hepatocellular carcinoma, gallbladder cancer, bronchial carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), multiple myeloma (MM), basal The tumor is selected from the group consisting of carcinoma, teratoma, retinoblastoma, choroidal melanoma, seminoma, rhabdomyosarcoma, osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma, Ewing's sarcoma, and plasmacytoma.

[0110] In other embodiments, the cancer is selected from the group consisting of lung cancer, non-small cell lung cancer, bronchioloalveolar carcinoma, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, cancer of the stomach, gastric cancer, colon cancer, breast cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal gland cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvis cancer, mesothelioma, hepatocellular carcinoma, biliary tract cancer, chronic or acute leukemia, lymphocytic lymphoma, tumors of the central nervous system (CNS), spinal axis tumors, brain stem glioma, glioblastoma multiforme, astrocytoma, Schwannoma, Selected from ependymoblastoma, medulloblastoma, meningioma, squamous cell carcinoma, and pituitary adenoma, including resistant and / or refractory forms of any of the above cancers, and one or more of the above Includes combinations of

[0111] In some embodiments, the cancer is selected from the group consisting of B-cell acute lymphocytic leukemia (B-ALL), mesothelioma, lymphoma, pancreatic cancer, lung cancer, gastric cancer, esophageal cancer, bladder cancer, brain cancer, head and neck cancer, melanoma, and breast cancer.

[0112] In other embodiments, the lung cancer is non-small cell lung cancer, small cell lung cancer, adenocarcinoma of the lung, and squamous cell carcinoma of the lung.

[0113] In other embodiments, the breast cancer is triple-negative breast cancer (TNBC).

[0114] In other embodiments, the brain tumor is a brain tumor selected from the group consisting of glioma, glioblastoma, astrocytoma, meningioma, medulloblastoma, peripheral neuroectodermal tumor, and craniopharyngioma.

[0115] In yet other embodiments, the cancer is a lymphoma selected from the group consisting of mantle cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, Burkitt's lymphoma, diffuse large B-cell lymphoma (DLBCL), and adult T-cell leukemia / lymphoma (ATLL). As used herein, the term adult T-cell leukemia / lymphoma refers to a rare and often aggressive T-cell lymphoma that can be found in the blood (leukemia), lymph nodes (lymphoma), skin, or multiple areas of the body.

[0116] As outlined above, methylthioadenosine phosphorylase (MTAP) is an enzyme found in all normal tissues that catalyzes the conversion of methylthioadenosine (MTA) to adenine and 5-methylthioribose-1-phosphate. Adenine is recycled to generate adenosine monophosphate, and 5-methylthioribose-1-phosphate is converted to methionine and glutamic acid. Because of this salvage pathway, MTA is not easily converted into metabolically active compounds such as L-alanosine. Antagonists can be used to serve as an alternative purine source when de novo purine synthesis is blocked. Many human and mouse malignant cells lack MTAP activity. MTAP deficiency is not only present in tissue culture cells, but the deficiency is also present in primary leukemia, glioma, melanoma, pancreatic cancer, non-small cell lung cancer (NSCLC), bladder cancer, astrocytoma, osteosarcoma, and head and neck cancer. MAT2A expression is also present in myxoid chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, soft tissue sarcoma, non-Hodgkin's lymphoma, and mesothelioma. For example, the proliferation of MTAP-null, i.e., MTAP-deficient, cancer cells is associated with increased MAT2A expression, as confirmed by the use of small molecule inhibitors of MAT2A, shRNAs. K. Marjon et al., Cell Reports 15 (2016) 574-587, which is incorporated herein by reference. MTAP-null or MTAP-deficient cancers are cancers in which the MTAP gene is deleted, lost, or otherwise inactivated, or in which the function or presence of the MTAP protein is reduced or impaired.

[0117] Certain embodiments of the present disclosure also provide methods of treating cancer in a subject, wherein the cancer is caused by a tumor in which the MTAP gene and / or MTAP protein is present and fully functional. The method is characterized by reduced or absent MTAP expression, or the absence of the MTAP gene, or reduced function of the MTAP protein, compared to cancers with a wild-type MTAP gene. The method comprises administering to a subject a compound of general formula I, or a pharmaceutically acceptable salt thereof, This involves administering a therapeutically effective amount of a tautomer, and / or isotopologue.

[0118] In another embodiment, there is provided a method for treating MTAP-deficient cancer in a subject, comprising administering to the subject an effective amount of a compound of general formula I, or a pharmaceutically acceptable salt thereof. In one embodiment, the MTAP-deficient cancer is leukemia, glioma, melanoma, pancreatic cancer, non-small cell lung cancer (NSCLC), bladder cancer, astrocytoma, osteosarcoma, head and neck cancer, myxoid chondrosarcoma, or ovarian cancer. , endometrial cancer, breast cancer, soft tissue sarcoma, lymphoma, and mesothelioma.

[0119] In one embodiment, the MTAP-deficient cancer is pancreatic cancer. In another embodiment, the MTAP-deficient cancer is bladder cancer, melanoma, brain cancer, lung cancer, pancreatic cancer, breast cancer, liver cancer, esophageal cancer, gastric cancer, colorectal cancer, head and neck cancer, renal cancer, colon cancer, diffuse large B-cell lymphoma (DLBCL), acute lymphoblastic leukemia (ALL), mantle cell lymphoma (MCL), glioblastoma multiforme (GBM), and and non-small cell lung cancer (NSCLC).

[0120] Genomic analysis of MTAP-null cell lines has revealed that cell lines incorporating mutant KRAS or p53 mutations are sensitive to MAT2A inhibition. Accordingly, an embodiment of the present disclosure is a method of treating cancer in a subject, wherein the cancer is characterized by reduced or absent MTAP expression, or the absence of the MTAP gene, or reduced function of the MTAP protein, the method comprising administering to the subject a therapeutically effective amount of a compound of general formula I, wherein the cancer is characterized by mutant KRAS or p53 mutations. The subject is further characterized by the presence of mutant p53. A method for treating MTAP-null cancers with a mutant KRAS or mutant p53 is provided, comprising: The method comprises administering to a subject a compound of general formula I, or a pharmaceutically acceptable salt, tautomer, For example, the cancer may be treated with an MTAP nuclease inhibitor or an isotopologue thereof. MTAP null and KRAS mutant, or MTAP null and p53 mutant, or MTAP null, KRAS mutant and p53 mutant.

[0121] The term "mutant KRAS" or "KRAS mutation" refers to a KRAS protein that incorporates an activating mutation that alters its normal function, and the gene encoding such a protein. For example, a mutant KRAS protein may incorporate a single amino acid substitution at position 12 or 13. In certain embodiments, the KRAS mutant incorporates a G12X or G13X substitution, where X represents any amino acid change at the indicated position. In certain embodiments, the substitution is G12V, G12R, G12C, or G13D. In another embodiment, the substitution is G13D. "Mutant p53" or "p53 mutation" refers to a p53 protein (or the gene encoding said protein) that incorporates a mutation that inhibits or eliminates its tumor suppressor function. In one embodiment, the p53 mutations are Y126_splice, K132Q, M133K, R174fs, R175H, R196*, C238S, C242Y, G245S, R248W, R248Q, I255T, D259V, S261_splice, R267P, R273C, R282W , A159V, or R280K. In certain embodiments, the cancer is non-small cell lung cancer (NSCLC), pancreatic cancer, head and neck cancer, gastric cancer, breast cancer, colorectal cancer, or ovarian cancer.

[0122] In another embodiment, the compounds disclosed herein are useful as ligands for the degradation of disease-related proteins. An example of this approach is PROTEOLYSIS TARGETING CHIMERAS (PROTEOLYSIS TARGETING CHIMERAS). PROTACs are bifunctional molecules that contain both a ligand portion capable of binding a target protein, selected from one of the compounds disclosed herein, and a ligase targeting portion, such as a peptide portion (called a degron) that is recognized and polyubiquitinated by an E3 ligase. Thus, PROTACs non-covalently bind to a target protein and recruit an E3 ligase via the degron, resulting in polyubiquitination and degradation of the bound target. Numerous publications describe the preclinical use of PROTACs in various therapeutic areas, including oncology. See, for example, Lu et al. Chemistry & Biology 22 (2015) 755-763. sea ​​bream.

[0123] Aspects

[0124] Aspect 1 A compound according to the following general formula I: [ka] During the ceremony, L is O, S, NR, or a bond; R is H or C1-C6 alkyl; R 1 is selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C3-C6 carbocyclyl, -(C1-C6 alkyl)(C3-C6 carbocyclyl), and -(C1-C6 alkyl)(C3-C6 cycloalkenyl). wherein: R 1 Any alkyl in is linear or branched; R 1 is optionally substituted with 1 to 6 halo; or, if L is NR, R and R in combination with L1 is one or more R A Replaced with 3-6 membered heterocycloalkyl (wherein 1-4 members of the ring are N, O, and S) (independently selected from R 2 and R 3 is C6-C 10 Aryl, C3-C6 carbocyclyl, 5-10 membered heteroaryl (wherein 1-4 members of the heteroaryl are independently selected from N, O, and S), and 3-14 membered heterocycloalkyl (wherein 1-4 members of the heterocycloalkyl are independently selected from N, O, and S). independently selected from the group consisting of: In the formula, R 2 and R 3 is R A , OR A , halo, -N=NR A , N.R. A R B , -(C1-C6 alkyl)NR A R B , -C(O)OR A , -C(O)NR A R B , -OC(O)R A and -CN, R 4 is H, C1-C6 alkyl (optionally substituted with one or more halo), -O(C1-C6 alkyl) (optionally substituted with one or more halo), -OH, halo, -CN, -(C1-C6 alkyl) Kill)NR A R B , and -NR A R B is selected from the group consisting of R 5 is H, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, ha -CN, and -NR C R D is selected from the group consisting of R A and R B is H, -CN, -hydroxy, oxo, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, -NH2, -S(O) 0-2 -(C1-C6 alkyl), -S(O) 0-2 -(C6-C 10 a -C(O)(C1-C6 alkyl), -C(O)(C3-C 14 carbocyclyl), -C3-C 14 Carbocycline -(C1-C6 alkyl)(C3-C 14 carbocyclyl), C6-C 10 Aryl, 3- to 14-membered heterocycloalkyl, and -(C1-C6 alkyl)-(3- to 14-membered heterocycloalkyl), where 1-4 members of the heterocycloalkyl are independently selected from N, O, and S, and 5- to 10-membered heteroaryl, where 1-4 members of the heteroaryl are independently selected from N, O, and S. ) independently selected from the group consisting of In the formula, R A and R B Each alkyl, alkoxy, alkenyl, alkynyl, aryl, carbocyclyl, heterocycloalkyl, and heteroaryl moiety in 10 Aryl, 3- to 14-membered heterocycloalkyl, and -(C1-C6 alkyl)-(3- to 14-membered heterocycloalkyl), where 1-4 ring members are independently selected from N, O, and S, and and 5-10 membered heteroaryl (wherein 1-4 members of the heteroaryl are independently selected from N, O, and S), —NHC(O)(OC-C alkyl), —NO, —CN, oxo, —C(O)OH, —C(O)O(C-C alkyl), —C-C alkyl(C-C alkoxy), —C(O)NH, C-C alkyl, —C(O)C-C alkyl, —OC-C alkyl, —Si(C-C alkyl), —S(O) 0-2 -(C1-C6 alkyl), C6-C 10 Aryl, -(C1-C6 alkyl)(C6-C 10 aryl), 3-14 membered heterocycloalkyl, and -(C-C alkyl)-(3-14 membered heterocycle), where 1-4 members of the heterocycle are independently selected from N, O, and S, and -O(C-C 14 aryl), wherein R A and R B Each alkyl, alkenyl, aryl, and heterocycloalkyl are optionally substituted with one or more substituents selected from the group consisting of hydroxy, -OC1-C6 alkyl, halo, -NH2, -(C1-C6-alkyl)NH2, -C(O)OH, CN, and oxo; R C and R D are each independently selected from H and C1-C6 alkyl, or a pharmaceutically acceptable salt thereof.

[0125] Aspect 2 2. A compound according to aspect 1, wherein: R 4 is H, C1-C6 alkyl optionally substituted with one or more halo, -O(C1-C6 alkyl) , -(C1-C6 alkyl)NR A R B , and -NR A R B (where R A and R Bis H and C1-C6 alkyl and R 5 is H, C1-C6 alkyl, C1-C6 alkoxy, and -NR C R D selected from the group consisting of compound.

[0126] Aspect 3 R 4 and R 5 is H.

[0127] Aspect 4 R 4 A compound according to any one of aspects 1 to 3, wherein is H.

[0128] Aspect 5 R 5 A compound according to any one of aspects 1 to 4, wherein is H.

[0129] Aspect 6 R 4 and R 5 A compound according to any one of aspects 1 to 5, wherein each of

[0130] Aspect 7 R 2 But C6-C 10 aryl or 5-10 membered heteroaryl. The compound according to any one of claims 1 to 5.

[0131] Aspect 8 R 2 But C 6- C 10 The compound according to embodiment 7, wherein the compound is aryl.

[0132] Aspect 9 R 2 is phenyl.

[0133] Aspect 10 R 2 is a 5-10 membered heteroaryl and one ring member is N.

[0134] Aspect 11 R 2 is 5- or 6-membered heteroaryl.

[0135] Aspect 12 R 2 12. The compound according to embodiment 10 or 11, wherein is a 6-membered heteroaryl.

[0136] Aspect 13 R 2 13. The compound according to any one of aspects 10 to 12, wherein is pyridyl.

[0137] Aspect 14 R 3 is a 3- to 14-membered heterocycloalkyl or a 5- to 10-membered heteroaryl. The compound according to any one of claims 1 to 12.

[0138] Aspect 15 R 3 is selected from the group consisting of benzothiazolyl, benzisothiazolyl, benzoxazolyl, pyridinyl, pyridinonyl, pyradazinyl, benzimidazolyl, benzotriazolyl, indazolyl, quinoxalinyl, quinolinyl, quinazolinyl, imidazopyridinyl, pyrazolopyridinyl, triazolopyridinyl, cinnolinyl, isoxazolyl, pyrazolyl, benzofuranyl, dihydrobenzofuranyl, dihydrobenzodioxinyl, and tetrahydrobenzodioxinyl.

[0139] Aspect 16 R 3 But C6-C 10 A compound according to any one of aspects 1 to 12, wherein the compound is aryl.

[0140] Aspect 17 R 3is phenyl.

[0141] Aspect 18 R 2 is phenyl and R 3 A compound according to any one of aspects 1 to 6, wherein is 3-14 membered heterocycloalkyl or 5-10 membered heteroaryl.

[0142] Aspect 19 Aspect 19. A compound according to any one of aspects 1 to 18, wherein L is O or NR.

[0143] Aspect 20 R 1 But C 1- C6 alkyl or C 3- 20. The compound according to embodiment 19, wherein the compound is a C6 carbocyclyl.

[0144] Aspect 21 R 1 is C1-C3 alkyl optionally substituted with 1 to 3 F.

[0145] Aspect 22 2. A compound according to aspect 1, wherein: L is O or NR and R is H, and R 1 is C1-C3 alkyl optionally substituted with 1 to 3 F, R 2 is a 3- to 14-membered heterocycloalkyl or a 5- to 10-membered heteroaryl (wherein one member of the heterocycloalkyl or heteroaryl is N) or C-C 10 is aryl, R 3 is a 3- to 14-membered heterocycloalkyl or a 5- to 10-membered heteroaryl, wherein 1-3 members of the heterocycloalkyl or heteroaryl are independently selected from N, O, and S. ,Also R 4 and R 5 Each of the is H, compound.

[0146] Aspect 23 23. The compound of embodiment 22, wherein L is NR.

[0147] Aspect 24 The compound according to embodiment 1, wherein the compound is selected from the table below: [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4] [Table 3-5] [Table 3-6] [Table 3-7] [Table 3-8] [Table 3-9] [Table 3-10] [Table 3-11]

[0148] Aspect 25 A therapeutically effective amount of a compound according to any one of aspects 1 to 24 or a pharmaceutically acceptable salt thereof. A pharmaceutical composition comprising the compound of claim 1, a compound of claim 1, a compound of claim 2, a compound of claim 3, a compound of claim 4, a compound of claim 5, a compound of claim 6, a compound of claim 7, a compound of claim 8, a compound of claim 9, a compound of claim 10, a compound of claim 11, a compound of claim 12, a compound of claim 13, a compound of claim 14, a compound of claim 15, a compound of claim

[0149] Aspect 26 A method of treating cancer in a subject suffering from cancer, comprising administering to the subject a method according to any one of aspects 1 to 24. A method comprising administering an effective amount of a MAT2A inhibitor compound described in any one of the above.

[0150] Aspect 27 27. The method of embodiment 26, wherein the cancer is an MTAP-deficient cancer.

[0151] Aspect 28 A method for inhibiting the synthesis of S-adenosylmethionine (SAM) in a cell, comprising administering to the cell an effective amount of a compound according to any one of aspects 1 to 24 or a pharmaceutically acceptable salt thereof 20. A method comprising:

[0152] Aspect 29 The method of embodiment 28, wherein the cell is in a subject.

[0153] Aspect 30 A method for inhibiting synthesis of S-adenosylmethionine (SAM) in a subject, comprising administering to the subject and administering an effective amount of at least one compound or salt thereof according to any one of aspects 1 to 24. The method includes providing.

[0154] Aspect 31 A method of treating cancer in a subject suffering from cancer, comprising administering to the subject a method according to any one of aspects 1 to 24. A method comprising administering an effective amount of a compound according to any one of the preceding claims.

[0155] Aspect 32 32. The method of embodiment 31, wherein the cancer is an MTAP-deficient cancer.

[0156] Aspect 33 Cancers include mesothelioma, neuroblastoma, rectal cancer, colon cancer, familial adenomatous polyposis cancer and hereditary non-polyposis colon cancer, esophageal cancer, labial cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, stomach cancer, adenocarcinoma, medullary thyroid cancer, papillary thyroid cancer, kidney cancer, renal parenchymal cancer, ovarian cancer, cervical cancer, uterine cancer, endometrial cancer, choriocarcinoma, pancreatic cancer, prostate cancer, bladder cancer, testicular cancer, breast cancer, and urinary tract cancer. 33. The method of any one of aspects 26, 27, 31, and 32, wherein the tumor is selected from the group consisting of: carcinoma, melanoma, brain tumor, lymphoma, head and neck cancer, acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), hepatocellular carcinoma, gallbladder cancer, bronchial carcinoma, small cell lung cancer, non-small cell lung cancer, multiple myeloma, basal cell carcinoma, teratoma, retinoblastoma, choroidal melanoma, seminoma, rhabdomyosarcoma, osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma, Ewing's sarcoma, and plasmacytoma.

[0157] Aspect 34 33. The method of embodiment 31 or 32, wherein the cancer is selected from the group consisting of B-cell acute lymphocytic leukemia (B-ALL), mesothelioma, lymphoma, pancreatic cancer, lung cancer, gastric cancer, esophageal cancer, bladder cancer, brain tumor, head and neck cancer, melanoma, and breast cancer.

[0158] Aspect 35 35. The method of embodiment 34, wherein the cancer is selected from the group consisting of lung cancer, non-small cell lung cancer, small cell lung cancer, adenocarcinoma of the lung, and squamous cell carcinoma of the lung.

[0159] Aspect 36 35. The method of embodiment 34, wherein the cancer is a brain tumor selected from the group consisting of glioma, glioblastoma, astrocytoma, meningioma, medulloblastoma, peripheral neuroectodermal tumor, and craniopharyngioma.

[0160] Aspect 37 35. The method of embodiment 34, wherein the cancer is triple-negative breast cancer (TNBC).

[0161] Aspect 38 Cancer consists of mantle cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, Burkitt lymphoma, diffuse large B-cell lymphoma, and adult T-cell leukemia / lymphoma. 35. The method of embodiment 34, wherein the lymphoma is selected from the group consisting of:

[0162] Aspect 39 A method of treating cancer in a subject suffering from cancer, the method comprising: detecting a cancer in which methylthioadenosine phosphorylase (MTAP) gene expression is reduced or absent compared to a cancer in which the MTAP gene or MTAP protein is present and / or fully functional; 25. The method of claim 24, wherein the compound of claim 1 or a pharmaceutically acceptable salt thereof is a compound of claim 1 or a pharmaceutically acceptable salt thereof. A method comprising administering a therapeutically effective amount to a subject.

[0163] Aspect 40 A compound according to any one of aspects 1 to 24, or a pharmaceutically acceptable salt thereof, for inhibiting the synthesis of S-adenosylmethionine (SAM).

[0164] Aspect 41 25. A method for treating cancer in a subject suffering from cancer, comprising administering to a subject a compound according to any one of aspects 1 to 24. or a pharmaceutically acceptable salt thereof.

[0165] Aspect 42 42. The compound of embodiment 41, wherein the cancer is an MTAP-deficient cancer.

[0166] Aspect 43 Cancers include mesothelioma, neuroblastoma, rectal cancer, colon cancer, familial adenomatous polyposis cancer and hereditary non-polyposis colon cancer, esophageal cancer, labial cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, stomach cancer, adenocarcinoma, medullary thyroid cancer, papillary thyroid cancer, kidney cancer, renal parenchymal cancer, ovarian cancer, cervical cancer, uterine cancer, endometrial cancer, choriocarcinoma, pancreatic cancer, prostate cancer, bladder cancer, testicular cancer, breast cancer, and urinary tract cancer. 43. The compound of embodiment 41 or 42, wherein the compound is selected from the group consisting of: carcinoma, melanoma, brain tumor, lymphoma, head and neck cancer, acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), hepatocellular carcinoma, gallbladder cancer, bronchial carcinoma, small cell lung cancer, non-small cell lung cancer, multiple myeloma, basal cell carcinoma, teratoma, retinoblastoma, choroidal melanoma, seminoma, rhabdomyosarcoma, osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma, Ewing's sarcoma, and plasmacytoma.

[0167] Aspect 44 43. The compound of embodiment 41 or 42, wherein the cancer is selected from the group consisting of B-cell acute lymphocytic leukemia (B-ALL), mesothelioma, lymphoma, pancreatic cancer, lung cancer, gastric cancer, esophageal cancer, bladder cancer, brain tumor, head and neck cancer, melanoma, and breast cancer.

[0168] Aspect 45 45. The compound of embodiment 44, wherein the cancer is selected from the group consisting of lung cancer, non-small cell lung cancer, small cell lung cancer, adenocarcinoma of the lung, and squamous cell carcinoma of the lung.

[0169] Aspect 46 45. The compound of embodiment 44, wherein the cancer is triple-negative breast cancer (TNBC).

[0170] Aspect 47 45. The compound of embodiment 44, wherein the cancer is a brain tumor selected from the group consisting of glioma, glioblastoma, astrocytoma, meningioma, medulloblastoma, peripheral neuroectodermal tumor, and craniopharyngioma.

[0171] Aspect 48 44. The compound of any one of aspects 41-43, wherein the cancer is a lymphoma selected from the group consisting of mantle cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, Burkitt's lymphoma, diffuse large B-cell lymphoma (DLBCL), and adult T-cell leukemia / lymphoma. [Example]

[0172] The present disclosure will be more fully understood by reference to the following examples, which should not, however, be construed as limiting the scope of the disclosure.

[0173] [Table 4]

[0174] [Table 5]

[0175] [Table 6-1] [Table 6-2]

[0176] General Experiment

[0177] In the following examples, reagents and solvents were obtained from commercial sources (e.g., Alfa, Acros, Sigma). The compounds were purchased from various suppliers (e.g., Aldrich, TCI, and Shanghai Chemical Reagent Company) and used without further purification unless otherwise specified. Flash chromatography was performed using a column equipped with 200-300 mesh silica gel particles with Ez Purifier III. Analytical and preparative thin-layer chromatography (TLC) plates were HSGF254 (thickness: 0.15-0.2 mm, Shanghai Anbang Company, China). Nuclear magnetic resonance (NMR) Spectra were acquired on a Brucker AMX-400 NMR (Brucker, Switzerland). Chemical shifts are reported in parts per million (ppm, δ) downfield from tetramethylsilane. Mass spectra The sample was analyzed by electrospray ionization (ESI) using a Waters LCT TOF mass spectrometer (Waters, USA). HPLC chromatography was performed using an Agilent 1200 Liquid Chromatography (Agilent, USA; column: Ultimate 4.6 mm × 50 mm, 5 μm; mobile phase A: 0.1% formic acid). Aqueous solutions were recorded in 1000 mL of ethanol (mobile phase B: acetonitrile). Microwave reactions were performed on an Initiator 2.5 Microwave Synthesizer (Biotage, Sweden).

[0178] General Procedure I: [ka]

[0179] Compounds of structure 1.9, 1.11, and 1.12 were obtained through the scheme illustrated as General Procedure I. Electrophilic bromination was used to generate heterocyclic bromide 1.2 from pyridazinone 1.1. The desired R3 group was introduced using Chan-Lam coupling to generate compound 1.3. In this reaction, the desired R4 and R5 groups were introduced using either Suzuki cross-coupling with reagent 1.4 (Method A) or Stille cross-coupling with reagent 1.5 (Method B). The resulting intermediate 1.6 was then cyclized to the desired bicyclic core 1.7 under basic conditions. The desired R2 group was introduced using Suzuki cross-coupling to generate compound 1.8. Compound 1.8 was alkylated to introduce the desired R1 group, giving final compounds of structure 1.9. Alternatively, compound 1.8 was chlorinated to give aryl chloride 1.10, which could then introduce the desired R1 group via nucleophilic aromatic substitution. Alternatively, compound 1.8 was activated using BOP to give the final compound of structure 1.11 (Method C). Activation and introduction of the desired N-linked R1 via nucleophilic aromatic substitution afforded final compounds of structure 1.12 (Method D).

[0180] Preparation of Example 101 by General Procedure I (Method A): [ka]

[0181] Step A: 5-Amino-6-bromo-4-chloropyridazin-3(2H)-one

[0182] To a suspension of 5-amino-4-chloropyridazin-3(2H)-one (1.2 g, 8.2 mmol, 1.0 equiv) and NaOAc (0.74 g, 9.1 mmol, 1.1 equiv) in MeCN (40 mL) was added Br (1.45 g, 9.1 mmol, 1.1 equiv) via syringe at 80 °C for 5 min. The resulting mixture was stirred at 80 °C for an additional 1 h. After cooling to room temperature, the volatiles were removed under reduced pressure and diluted with ice-cold HO (20 mL). The resulting The resulting white precipitate was filtered, and the filter cake was collected and dried under reduced pressure to give 5-amino-6-bromo-4-chloropyridazin-3(2H)-one (1.48 g, 80% yield) as a white solid. LC-MS (ESI): m / z 224, 226 [M+H] + .

[0183] Step B: 5-amino-6-bromo-4-chloro-2-(2-methyl-2H-indazol-5-yl)pyridazin-3(2H)-one

[0184] To a suspension of 5-amino-6-bromo-4-chloropyridazin-3(2H)-one (590 mg, 2.63 mmol, 1.0 equiv) in DMF (15 mL) was added (2-methyl-2H-indazol-5-yl)boronic acid (555 mg, 3.15 mmol, 1.2 equiv), Cu(OAc) (478 mg, 2.63 mmol, 1.0 equiv) and pyridine (422 μL, 5.26 mmol, 2.0 The resulting mixture was stirred at 50°C (air atmosphere) for 8 hours, and the reaction was monitored by TLC. After completion, the reaction mixture was diluted with H2O (30 mL) and the resulting suspension was stirred for an additional 30 min. After stirring for 1 h, the precipitate was collected, washed with ice-cold HO (30 mL × 3), and dried under reduced pressure to give 5-amino-6-bromo-4-chloro-2-(2-methyl-2H-indazol-5-yl)pyridazin-3(2H)-one (770 mg, 82% yield) as an off-white solid. LC-MS (ESI): m / z 354, 356 [M+H] + .

[0185] Step C: Ethyl (E)-3-(4-amino-5-chloro-1-(2-methyl-2H-indazol-5-yl)-6-oxo-1,6-dihydropyridazin-3-yl)acrylate

[0186] 5-Amino-6-bromo-4-chloro-2-(2-methyl-2H-indazol-5-yl)piperidin in DMF (10 mL) To a solution of ridazin-3(2H)-one (500 mg, 1.41 mmol, 1.0 equiv.), ethyl (E)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)acrylate (350 mg, 1.55 mmol, 1.1 equiv.), Pd(dppf)Cl (103 mg, 0.14 mmol, 0.1 equiv.) and KCO (389 mg, 2.82 mmol, 2.0 equiv.) was added. The reaction mixture was stirred at 80° C. for 16 hours under a N atmosphere. The reaction mixture was poured into ice water (30 mL) and extracted with EtOAc (50 mL×3). The combined organic layers were dried over NaSO and concentrated under reduced pressure, and the crude residue was purified by flash column chromatography on silica gel to give ethyl (E)-3-(4-amino-5-chloro-1-(2-methyl-2H-indazol-5-yl)-6-oxo-1,6-dihydropyridazin-3-yl)acrylate (316 mg, 60% yield) as a yellow solid. LC-MS (ESI): m / z 374 [M+H] + .

[0187] Step D: 4-chloro-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione

[0188] Ethyl (E)-3-(4-amino-5-chloro-1-(2-methyl-2H-indazol-5-yl)methyl ... To a solution of (1,6-dihydropyridazin-3-yl)-6-oxo-1,6-dihydropyridazin-3-yl acrylate (400 mg, 1.07 mmol, 1.0 equiv.) was added K2CO3 (295 mg, 2.14 mmol, 2.0 equiv.) at room temperature. The reaction mixture was heated at 80 °C for 3 h. The mixture was stirred for 1 hour. Ice water (30 mL) was then added, and the mixture was extracted with EtOAc (30 mL × 3). The combined organic layers were dried over NaSO and concentrated under reduced pressure, and the crude residue was purified by flash column chromatography on silica gel to give 4-chloro-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione (319 mg, 85% yield) as a yellow solid. LC-MS (ESI): m / z 328 [M+H] + .

[0189] Step E: 4-(4-chlorophenyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione

[0190] 4-Chloro-2-(2-methyl-2H-indazole-5- (yl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione (1 g, 3.05 mmol, 1.0 equiv.), (4-chlorophenyl)boronic acid (954 mg, 6.1 mmol, 2.0 equiv.), Pd(OAc) (68 mg, 0.3 mmol, 0.1 equiv.), S-Phos (251 mg, 0.61 mmol, 0.2 equiv.), and KCO (1.26 g, 9.15 mmol, 3.0 equiv.). The solution was stirred at 110° C. under a N 2 atmosphere for 2 h. Ice water (30 mL) was added and the mixture was extracted with EtOAc (30 mL × 3), the combined organic layers were dried over Na2SO4 and concentrated under reduced pressure, and the crude residue was purified by flash column chromatography on silica gel to give 4-(4- 700 mg of (2-chlorophenyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione (57% yield) was obtained. LC-MS: m / z 404 [M+H] + .

[0191] Step F: 4-(4-chlorophenyl)-6-(cyclopropylmethoxy)-2-(2-methyl-2H-yne)- (dazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one

[0192] A solution of 4-(4-chlorophenyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione (200 mg, 0.495 mmol, 1.0 equiv), (bromomethyl)cyclopropane (0.2 mL, 2.0 mmol, 4.0 equiv), and CsCO (484 mg, 1.5 mmol, 3.0 equiv) in DMF (3 mL) was stirred at room temperature for 16 h. The reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by preparative HPLC to give 4-(4-chlorophenyl)-6-(cyclopropylmethoxy)-2-(2-methyl-2H-indazol-5-yl). (2H)-pyrido[3,2-c]pyridazin-3(2H)-one (Example 101).

[0193] 1 H NMR (400 MHz, DMSO-d6) δ: 8.55 (s, 1H), 8.07 (d, J = 2.0 Hz, 1H), 8.04 (d, J = 9.6 Hz, 1H), 7.86 (d, J = 8.8 Hz, 2H), 7.76 (d, J = 9.2 Hz, 1H), 7.56 (d, J = 8.4 Hz, 2H), 7.51 (dd, J = 9.2 Hz, 1.6 Hz, 1H), 7.04 (d, J = 9.2 Hz, 1H), 4.28 (s, 3H), 4.21 (d, J = 7.2 Hz, 2H), 1.36-1.19 (m, 1H), 0.64-0.57 (m, 2H), 0.37-0.33 (m, 2H).

[0194] LC-MS (ESI): m / z 458 [M+H] +

[0195] Preparation of Example 102 by General Procedure I (Method B): [ka]

[0196] Step C: 4-chloro-6-hydroxy-2-(2-methyl-2H-indazol-5-yl)-8-(trifluoromethyl)pyrido[3,2-c]pyridazin-3(2H)-one

[0197] 5-Amino-6-bromo-4-chloro-2-(2-methyl-2H-indazol-5-yl) in DMF (10 mL) Pyridazin-3(2H)-one (1 g, 2.8 mmol, 1.0 equiv.), ethyl (Z)-4,4,4-trifluoro-3-(trifluoromethyl)- A solution of (tributylstannyl)but-2-enoate (see Synlett, 2012, 23, 755-759) (2.6 g, 5.6 mmol, 2.0 equiv.), Pd(PPh3)4 (655 mg, 0.567 mmol, 0.2 equiv.), and CuI (216 mg, 1.13 mmol, 0.4 equiv.) was stirred at 100 °C for 16 h under a N2 atmosphere. The reaction mixture was diluted with CsF (saturated The mixture was quenched with 30 mL of HCl (aqueous solution) and stirred for an additional 30 minutes. The resulting suspension was filtered and the precipitate was was collected and triturated with EtOAc (20 mL) to give 4-chloro-6-hydroxy-2-(2-methyl-2H-indazol-5-yl)-8-(trifluoromethyl)pyrido[3,2-c]pyridazine-3(2H)-o as a yellow solid The compound (800 mg, 71% yield) was obtained. LC-MS (ESI): m / z 396 [M+H] + .

[0198] 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(2-methyl-2H- Indazol-5-yl)-8-(trifluoromethyl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 102) was prepared by general procedure I (steps E, F) from 2-(4-(difluoromethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane and 2,2-difluoroethyltrifluoromethyl. It was synthesized from methylmethanesulfonate.

[0199] 1 H NMR (400 MHz, DMSO-d6) δ: 8.56 (s, 1H), 8.07 (s, 1H), 7.87 (d, J = 8.8 Hz, 2H), 7.76 (d, J = 9.2 Hz, 1H), 7.61 (s, 1H), 7.50 (dd, J = 9.2 Hz, 1.6 Hz, 1H), 7.38 (t, J HF = 73.6 Hz, 1H), 7.29 (d, J = 8.4 Hz, 2H), 6.43 (tt, J HF = 54.0 Hz, J = 2.8 Hz, 1H), 4.63 (td, J HF = 14.8 Hz, J = 2.8 Hz, 2H), 4.25 (s, 3H).

[0200] LC-MS (ESI): m / z 568 [M+H] + .

[0201] Preparation of Example 103 by General Procedure I (Method C): [ka]

[0202] Step G: 6-chloro-4-(4-chlorophenyl)-2-(2-methyl-2H-indazol-5-yl)piperidin Lido[3,2-c]pyridazin-3(2H)-one

[0203] A solution of 4-(4-chlorophenyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione (100 mg, 0.284 mmol, 1.0 equiv.) in POCl3 (5 mL) was heated at 80 °C for 2 h. The excess POCl3 was removed under reduced pressure, and the residue was carefully poured into ice-cold NaHCO3 (saturated aqueous solution) (10 mL). The resulting mixture was extracted with DCM (10 mL x 3). The combined organic layer was washed with water and stirred for 1 hour. Washed with water (10 mL), dried over Na2SO4, concentrated under reduced pressure, and the crude residue was purified by flash column chromatography on silica gel to give 6-chloro-4-(4-chlorophenyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazine-3(2H)- as a red solid The compound (60 mg, 57% yield) was obtained. LC-MS (ESI): m / z 422 [M+H] + .

[0204] Step H: 4-(4-chlorophenyl)-6-(ethylamino)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one

[0205] 6-chloro-4-(4-chlorophenyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (60 mg, 0.142 mmol, 1.0 equiv.), ethylamine hydrochloride in THF (3 mL). A solution of the salt (58 mg, 0.71 mmol, 5.0 equiv.), CsF (108 mg, 0.71 mmol, 5.0 equiv.), and DIPEA (92 mg, 0.71 mmol, 5.0 equiv.) was stirred at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure, and the crude residue was purified by preparative HPLC to give 4-(4-chlorophenyl)-6-(ethylamino)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 103).

[0206] 1 H NMR (400 MHz, DMSO-d6) δ: 8.46 (s, 1H), 8.34 (t, J = 5.2 Hz, 1H), 7.94 (d, J = 1.2 Hz, 1H), 7.82 (d, J = 8.4 Hz, 2H), 7.67 (d, J = 9.2 Hz, 1H), 7.61 (d, J = 9.2 Hz, 1H), 7.45-7.40 (m, 3H), 6.79 (d, J = 9.2 Hz, 1H), 4.22 (s, 3H), 3.37-3.30 (m, 2H), 1.15 (t, J = 7.2 Hz, 3H).

[0207] LC-MS (ESI): m / z 431 [M+H] + .

[0208] Preparation of Example 301 by General Procedure I (Method D): [ka]

[0209] Step I: 2-(2-methyl-2H-indazol-5-yl)-4-(6-methylpyridin-3-yl)-6-((2,2,2-trifluoroethyl)amino)pyrido[3,2-c]pyridazin-3(2H)-one (Example 301)

[0210] To a solution of 6-hydroxy-2-(2-methyl-2H-indazol-5-yl)-4-(6-methylpyridin-3-yl)pyrido[3,2-c]pyridazin-3(2H)-one (100 mg, 0.26 mmol, 1.0 equiv.), BOP (173 mg, 0.39 mmol, 1.5 equiv.) in DMF (5 mL) was added DIEA (0.13 mL, 0.78 mmol, 3.0 equiv.). The resulting mixture was stirred at room temperature for 30 minutes, and then 2,2,2-trifluoroethan-1-amine (0.06 mL, 0.78 mmol, 3.0 equiv.) was added and stirred at room temperature for an additional 2 hours. Upon completion, the reaction was quenched by adding ice-water (10 mL), extracted with EtOAc (10 mL × 3), dried over NaSO, and purified under reduced pressure. Upon concentration under reduced pressure, the residue was purified by flash column chromatography on silica gel to give 2-(2-methyl-2H-indazol-5-yl)-4-(6-methylpyridin-3-yl)-6-((2,2,2-trifluoroethyl)amino)pyrido[3,2-c]pyridazin-3(2H)-one (Embodiment 301). .

[0211] 1 H NMR (400 MHz, DMSO-d6) δ: 8.86 (s, 1H), 8.80 (s, 1H), 8.47 (s, 1H), 8.02 (d, J = 7.5 Hz, 1H), 7.96 (s, 1H), 7.77 (d, J = 9.5 Hz, 1H), 7.68 (d, J = 8.8 Hz, 1H), 7.42 (d, J = 9.7 Hz, 1H), 7.28 (d, J = 8.1 Hz, 1H), 6.92 (d, J = 9.6 Hz, 1H), 4.30-4.22 (m, 2H), 4.21 (s, 3H), 2.51 (s, 3H).

[0212] LC-MS (ESI): m / z 466 [M+H] + .

[0213] Using the procedure described above for General Procedure I (Method A), the following compounds were synthesized using the appropriate starting materials: [Table 7-1] [Table 7-2] [Table 7-3] [Table 7-4] [Table 7-5] [Table 7-6] [Table 7-7] [Table 7-8] [Table 7-9] [Table 7-10] [Table 7-11] [Table 7-12] [Table 7-13] [Table 7-14] [Table 7-15] [Table 7-16]

[0214] Using the procedure described above for General Procedure I (Method C), the following compounds were synthesized using the appropriate starting materials: [Table 8-1] [Table 8-2] [Table 8-3] [Table 8-4] [Table 8-5] [Table 8-6] [Table 8-7]

[0215] Using the procedure described above for General Procedure I (Method D), the following compounds were synthesized using the appropriate starting materials: [Table 9-1] [Table 9-2]

[0216] Synthesis of intermediate (6-(methyl-d3)pyridin-3-yl)boronic acid: [ka]

[0217] Step A: 5-Methoxy-2-(methyl-d3)pyridine

[0218] To a solution of 2-bromo-5-methoxypyridine (7 g, 37.2 mmol, 1.0 equiv.) and Fe(acac) (1.31 g, 3.71 mmol, 0.1 equiv.) in anhydrous THF (70 mL) was added CDMgI (1 M in THF) (93 mL, 93 mmol, 2.5 equiv.) dropwise at 0 °C under a N atmosphere, and the resulting mixture was stirred at 0 °C for 3 h. After completion, the reaction was quenched by adding NHCl (sat. aq.) (200 mL) and then extracted with EtOAc (70 mL × 3). The combined organic layers were washed with brine (100 mL), dried over Na2SO4 and concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel to give 5-methoxy-2-(methyl-d3)pyridine (4 g, 85%) as a colorless oil. LC-MS (ESI): m / z 127 [M+H] + .

[0219] Step B: 6-(methyl-d3)pyridin-3-ol

[0220] To a solution of 5-methoxy-2-(methyl-d3)pyridine (1.6 g, 12.6 mmol, 1.0 equiv.) in dry toluene (20 mL) was added L-Selectride (1 M in THF) (37.8 mL, 37.8 mmol, 3.0 equiv.) dropwise via addition funnel at 0° C. After addition, the reaction mixture was warmed to room temperature and transferred to a preheated oil bath (110° C.) After completion, the reaction was cooled to 0° C. again and quenched by careful addition of MeOH (10 mL). The resulting mixture was concentrated under reduced pressure and the residue was purified by flash column chromatography on silica gel to give 6-(methyl-d3)pi as a pale yellow solid. Lysin-3-ol (1.2 g, 84.3%) was obtained. LC-MS (ESI): m / z 113 [M+H] + .

[0221] Step C: 6-(methyl-d3)pyridin-3-yl trifluoromethanesulfonate

[0222] 6-(methyl-d3)pyridin-3-ol (500 mg, 4.46 mmol, 1.0 equiv) in dry DCM (10 mL) To a solution of 1,2-dichloromethane and pyridine (0.54 mL, 6.69 mmol, 1.5 equiv.) at 0° C. was added triflic anhydride (1.13 mL, 6.69 mmol, 1.5 equiv.) via syringe. The reaction mixture was allowed to warm to room temperature and then After completion, the reaction was quenched by adding HO (20 mL) and then extracted with EtOAc (20 mL × 3). The combined organic layers were washed with diluted HCl (0.5 N, aq.) (20 mL), brine (20 mL), dried over NaSO, concentrated under reduced pressure, and the residue was purified by flash filtration with silica gel. Purification by column chromatography gave 6-(methyl-d3)pyridin-3-yl trifluoromethanesulfonate (1.0 g, 91%) as a colorless oil. LC-MS (ESI): m / z 245 [M+H] + .

[0223] Step D: (6-(methyl-d3)pyridin-3-yl)boronic acid

[0224] To a solution of 6-(methyl-d3)pyridin-3-yl trifluoromethanesulfonate (1.0 g, 4.09 mmol, 1.0 equiv.) in dry 1,4-dioxane (10 ml), bis(pinacolato)diboron (2.08 g, 8.2 mmol, 2.0 equiv.), KOAc (1.6 g, 16.4 mmol, 4.0 equiv.), and Pd(dppf)Cl2 (300 mg, 0.41 mmol, 0.1 equiv.) were added. The resulting mixture was stirred at 100 °C under a N2 atmosphere for 16 h. Completion The crude mixture was then filtered through a short pad of Celite®, the filtrate was concentrated under reduced pressure and the residue was purified by RP preparative HPLC to give (6-(methyl-d3)pyridin-3-yl)boronic acid (460 mg, 80%). LC-MS (ESI): m / z 141 [M+H] + .

[0225] General Procedure II: [ka]

[0226] Compounds of structure 2.8 and 2.11 were obtained through the scheme shown as General Procedure II. Starting with pyridazinone 2.1, the heterocycle was benzylated to give compound 2.2. The desired R4 and R5 groups were introduced using Suzuki cross-coupling to give compounds of structure 2.3. Compound 2.3 was then cyclized under basic conditions to give bicyclic compound 2.4. The desired R2 group was introduced using Suzuki coupling to give compound 2.5. Compound 2.5 was then chlorinated to give aryl chloride 2.6. The desired R1 group was then added via nucleophilic aromatic substitution. The heterocyclic core was then simultaneously debenzylated to give compound 2.7. Finally, the desired R3 Groups are introduced using Ullmann coupling (Method A) or Chan-Lam coupling (Method B) Alternatively, compound 2.5 can be activated with BOP to afford the desired R1 amine. Reaction could afford heterocycle 2.9 (Method C). The benzyl group was removed using t-BuOK to give heterocycle 2.10, and the desired R group was introduced using Ullmann coupling to give compounds of structure 2.11.

[0227] General Procedure IIa (Method C): [ka]

[0228] General Procedure IIa (Method C) uses Br as an alternative protecting group to the Bn protecting group shown in General Procedure II (Method C). Deprotection of heterocycle 2.16 under acidic conditions led to the convergent synthesis of compound structure 2.11.

[0229] Preparation of Example 166 by General Procedure II (Method A): [ka]

[0230] Step A: 5-amino-2-benzyl-6-bromo-4-chloropyridazin-3(2H)-one

[0231] To a solution of 5-amino-6-bromo-4-chloropyridazin-3(2H)-one (3 g, 13.4 mmol, 1.0 equiv.), KCO (3.7 g, 26.8 mmol, 2.0 equiv.) in DMF (50 mL) was added BnBr (2.5 g, 14.7 mmol, 1.1 equiv.), and the reaction mixture was stirred at 80 °C overnight. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (50 mL × 3). The combined organic layers were washed with brine (50 mL), dried over NaSO, and concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel to give 5-amino-2-benzyl-6-bromo-4-chloropyridazin-3(2H)-one (2.46 g, 59% yield) as a white solid. LC-MS (ESI) m / z 314, 316 [M+H] +.

[0232] Step B: Ethyl (E)-3-(4-amino-1-benzyl-5-chloro-6-oxo-1,6-dihydropyridinyl) (dazin-3-yl)acrylate

[0233] 5-Amino-2-benzyl-6-bromo-4-chloropyridazin-3(2H)-one (2.46 g) in DMF (40 mL) To a solution of 4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl (E)-acrylate (1.94 g, 8.6 mmol, 1.1 equiv.), K2CO3 (2.2 g, 15.6 mmol, 2.0 equiv.), and ethyl (E)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)acrylate (1.94 g, 8.6 mmol, 1.1 equiv.), Pd(dppf)Cl2 (0.57 g, 0.8 mmol, 0.1 equiv.) was added under a N2 atmosphere, and the reaction mixture was stirred at 100 °C for 3 h. The reaction mixture was diluted with HO (50 mL) and extracted with EtOAc (50 mL × 3). The combined organic layers were washed with brine (50 mL), dried over NaSO, concentrated under reduced pressure, and the residue was purified by column chromatography on silica gel to give (E)-3-(4-amino-1-benzyl-5-chloro-6-oxo-1,6-dihydropyridazin-3-yl)acrylate (1.89 g, mp 172-174°C, 187-188°F) as a brown solid. The yield was 71%. LC-MS (ESI): m / z 334 [M+H] + .

[0234] Step C: 2-Benzyl-4-chloropyrido[3,2-c]pyridazine-3,6(2H,5H)-dione

[0235] Ethyl (E)-3-(4-amino-1-benzyl-5-chloro-6-oxo-1,6-dihydro)- To a stirred solution of (pyridazin-3-yl)acrylate (1.89 g, 5.66 mmol, 1.0 equiv.), K2CO3 (2.34 g, 16.98 mmol, 3.0 equiv.) was added, and the reaction mixture was stirred at 80 °C overnight. The reaction mixture was diluted with water (30 mL), extracted with EtOAc (30 mL × 3), and the combined organic layers were dried over Na2SO4 and concentrated under reduced pressure to give crude 2-benzyl-4-chloropyrido[3,2-c]pyridazine-3,6(2H,5H)-dione as a brown solid (1.5 g), which was used in the next step without further purification. LC-MS (ESI): m / z 288 [M+H] + .

[0236] Step D: 2-Benzyl-4-(4-(difluoromethoxy)phenyl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione

[0237] 2-Benzyl-4-chloropyrido[3,2-c]pyridin in a dioxane / HO mixture (88 mL, 10 / 1, v / v) Dazin-3,6(2H,5H)-dione (1.5 g, 5.2 mmol, 1.0 equiv.), 2-(4-(difluoromethoxy)phenyl)- To a stirred solution of (4,4,5,5-tetramethyl-1,3,2-dioxaborolane) (2 g, 7.2 mmol, 1.4 equiv.), K2CO3 (1.54 g, 11.1 mmol, 2.1 equiv.), and X-Phos (0.52 g, 1.1 mmol, 0.2 equiv.) was added a nitrogen atmosphere. Pd(OAc)2 (0.12 g, 0.55 mmol, 0.1 equiv.) was added under reduced pressure. The reaction mixture was stirred at 110 °C overnight. The reaction mixture was concentrated under reduced pressure and purified by column chromatography on silica gel to give 2-benzyl-4-(4-(difluoromethoxy)phenyl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione (1.4 g, 68% yield) as a white solid. LC-MS (ESI): m / z 396 [M+H] + .

[0238] Step E: 2-benzyl-6-chloro-4-(4-(difluoromethoxy)phenyl)pyrido[3,2-c] Pyridazin-3(2H)-one

[0239] 2-Benzyl-4-(4-(difluoromethoxy)phenyl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione (400 mg, 1.01 mmol, 1.0 equiv) was dissolved in POCl (4 mL), and the resulting mixture was stirred at 80 °C for 4 h. Excess POCl was removed under reduced pressure, and the residue was poured onto ice-cold NaHCO (saturated aqueous solution) (20 mL) and extracted with DCM (30 mL × 3). The combined organic layers were dried over NaSO and concentrated under reduced pressure to give crude 2-benzyl-6-chloro-4-(4-(difluoromethoxy)phenyl)pyrido[3,2-c]pyridazin-3(2H)-one (400 mg, 95% yield) as a yellow solid. LC-MS (ESI): m / z 414 [M+H] + .

[0240] Step F: 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)pyridine Do[3,2-c]pyridazin-3(2H)-one

[0241] 2-Benzyl-6-chloro-4-(4-(difluoromethoxy)phenyl)pyrido[3,2-c]pyridazin-3(2H)-one (400 mg, 0.97 mmol, 1.0 equiv.), 2,2-difluoroethane-1-ol in anhydrous THF (8 mL) To a solution of t-BuOK (541 mg, 4.8 mmol, 5.0 equiv.) in ethanol (396 mg, 4.8 mmol, 5.0 equiv.) was added t-BuOK (541 mg, 4.8 mmol, 5.0 equiv.) at 0 °C. The reaction mixture was added in several portions, and after the addition, the reaction mixture was stirred at room temperature overnight. The reaction mixture was diluted with H2O (10 mL), extracted with EtOAc (30 mL x 3), and the combined organic layers were dried over Na2SO4 and concentrated under reduced pressure. The mixture was concentrated with 500 ml of ethyl acetate and the residue was purified by flash column chromatography on silica gel to give 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)pyridine as a yellow solid. Do[3,2-c]pyridazin-3(2H)-one (150 mg, 42% yield) was obtained. LC-MS (ESI): m / z 370 [M+H] + .

[0242] Step G: 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(2,3-dimethyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Method A)

[0243] To a stirred suspension of 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)pyrido[3,2-c]pyridazin-3(2H)-one (70 mg, 0.19 mmol, 1.0 equiv.) in MeCN (3 mL) was added 5-bromo-2,3-dimethyl-2H-indazole (64.0 mg, 0.28 mmol, 1.5 equiv.), Cul (36.2 mg, 0.19 mmol, 1.0 equiv.), N 1 ,N 2 -Dimethylcyclohexane-1,2-diamine (26.9 mg, 0.19 mmol, 1.0 equiv.) and CsF (57.6 mg, 0.38 mmol, 2.0 equiv.) were added. The reaction was stirred overnight at 85° C. in a sealed tube under a N atmosphere and concentrated under reduced pressure. The residue was purified by preparative HPLC to give 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(2,3-dimethyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 166).

[0244] 1 H NMR (400 MHz, DMSO-d6) δ: 8.07 (d, J = 9.4 Hz, 1H), 7.98 (d, J = 1.4 Hz, 1H), 7.87 (d, J = 8.0 Hz, 2H), 7.62 (d, J = 8.0 Hz, 1H), 7.42 (dd, J = 9.1 Hz, 2.0 Hz, 1H), 7.35 (t, J HF= 72.0 Hz, 1H), 7.26 (d, J = 8.8 Hz, 2H), 7.06 (d, J = 9.4 Hz, 1H), 6.41 (tt, J HF = 54.4 Hz, J = 3.3 Hz, 1H), 4.59 (td, J HF = 15.1 Hz, J = 3.3 Hz, 2H), 4.10 (s, 3H), 2.64 (s, 3H).

[0245] LC-MS (ESI): m / z 514 [M+H] + .

[0246] Preparation of Example 167 by General Procedure II (Method B): [ka]

[0247] Step G: 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(1-methyl-1H-benzo[d]imidazol-6-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Method Law B)

[0248] 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)pyridinium chloride in DMF (5 mL) To a suspension of 1-methyl-1H-do[3,2-c]pyridazin-3(2H)-one (70 mg, 0.19 mmol, 1.0 equiv.) Benzo[d]imidazol-6-ylboronic acid (33.4 mg, 0.23 mmol, 1.2 equiv.), Cu(OAc) (34.5 mg, 0.19 mmol, 1.0 equiv.), and pyridine (30.0 mg, 0.38 mmol, 2.0 equiv.) were added. After stirring overnight at 50°C under air atmosphere, the reaction mixture was quenched by adding H2O (10 mL). The combined organic layer was dried over anhydrous Na2SO4 and concentrated under reduced pressure, and the residue was purified by preparative HPLC to give 6-(2,2-difluoroethoxy)-4-(4-(difluoroethoxy)- ... (fluoromethoxy)phenyl)-2-(1-methyl-1H-benzo[d]imidazol-6-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 167).

[0249] 1 H NMR (400 MHz, DMSO-d6) δ: 8.34 (s, 1H), 8.06 (d, J = 9.4 Hz, 1H), 7.92 (d, J = 1.8 Hz, 1H), 7.88 (d, J = 8.0 Hz, 2H), 7.79 (d, J = 8.6 Hz, 1H), 7.47 (dd, J = 8.6 Hz, 2.0 Hz, 1H), 7.35 (t, J HF = 72.0 Hz, 1H), 7.26 (d, J = 8.8 Hz, 2H), 7.07 (d, J = 9.4 Hz, 1H), 6.41 (tt, J HF = 54.4 Hz, 3.3 Hz, 1H), 4.60 (td, J HF = 15.1 Hz, 3.3 Hz, 2H), 3.88 (s, 3H).

[0250] LC-MS (ESI): m / z 500 [M+H] + .

[0251] Preparation of Example 321 by General Procedure II (Method C): [ka]

[0252] Step H: 2-benzyl-4-(6-(difluoromethyl)pyridin-3-yl)-6-((ethyl-d5) Amino)pyrido[3,2-c]pyridazin-3(2H)-one

[0253] 2-Benzyl-4-(6-(difluoromethyl)pyridin-3-yl)pyrido[3,2-c]pyridin in DMF (2 mL) To a solution of ridazine-3,6(2H,5H)-dione (250 mg, 0.66 mmol, 1.0 equiv.), BOP (436 mg, 0.99 mmol, 1.5 equiv.) and DIEA (584 μL, 3.29 mmol, 5.0 equiv.) were added, and the reaction mixture was stirred at room temperature for 1 h. After stirring, ethylamine-d5 hydrochloride (86 mg, 0.99 mmol, 1.5 equiv.) was added, and the resulting mixture The mixture was stirred at room temperature for an additional 0.5 h. Upon completion, the reaction was cooled to room temperature by adding ice water (10 mL). The mixture was quenched with hexane, extracted with EtOAc (10 ml × 3), dried over NaSO, concentrated under reduced pressure, and the residue was purified by flash column chromatography on silica gel to give 4-(6-(difluoromethyl)pyridin-3-yl)-6-((ethyl-d5)amino)pyridopyridazin-3(2H)-one (234 mg, 86%) as a yellow solid. LC-MS (ESI): m / z 413 [M+H] + .

[0254] Step I: 4-(6-(difluoromethyl)pyridin-3-yl)-6-((ethyl-d5)amino)pyrido[3,2-c]pyridazin-3(2H)-one

[0255] To a solution of 2-benzyl-4-(6-(difluoromethyl)pyridin-3-yl)-6-((ethyl-d5)amino)pyridopyridazin-3(2H)-one (185 mg, 0.45 mmol, 1.0 equiv) in THF / DMF (2 mL, 1 / 1) was added t-BuOK (251 mg, 2.25 mmol, 5.0 equiv) and the reaction mixture was stirred at 70° C. for 8 h. Afterwards, the pH was adjusted to about 7 by adding 1N HCl (aq), and then the mixture was extracted with DCM (10 mL × 3). The combined organic layers were washed with brine (20 mL), dried over NaSO, concentrated under reduced pressure, and the residue was purified by flash chromatography to give 4-(6-(difluoromethyl)pyridin-3-yl)-6-((ethyl-d5)amino)pyridopyridazin-3(2H)-one (50 mg) as a yellow solid. LC-MS (ESI): m / z 323 [M+H] + .

[0256] Step J: 4-(6-(difluoromethyl)pyridin-3-yl)-6-((ethyl-d5)amino)-2-(2-(methyl-d3)-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one

[0257] 4-(6-(difluoromethyl)pyridin-3-yl)-6-((ethyl-d5)amino)pyridin-3-yl in ACN (1 mL) To a solution of dopyridazin-3(2H)-one (50 mg, 0.16 mmol, 1.0 equiv) and 5-bromo-2-(methyl-d3)-2H-indazole (50 mg, 0.23 mmol, 1.5 equiv), CuI (30 mg, 0.16 mmol, 1.0 equiv), CsF (47 mg, 0.31 mmol, 2.0 equiv), N1,N2-dimethylchlorohexane-1,2-diamine (22 mg, 0.16 mmol, 1.0 equiv) were added and the reaction mixture was stirred at 85 °C for 14 h. After completion, the reaction mixture The mixture was diluted with HO (5 mL) and extracted with EtOAc (10 mL × 3). The combined organic layers were washed with brine (20 mL), dried over NaSO, concentrated under reduced pressure, and the residue was purified by RP preparative HPLC to give 4-(6-(difluoromethyl)pyridin-3-yl)-6-((ethyl-d5)amino)-2-(2-(methyl-d3)-2H-indazol-5-yl)pyridopyridazin-3(2H)-one (Example 321).

[0258] 1H NMR (400 MHz, DMSO-d6) δ (ppm): 9.06 (s, 1H), 8.46 (d, J = 2.6 Hz, 2H), 8.39 (d, J = 8.0 Hz, 1H), 7.95 (s, 1H), 7.73 (d, J = 8.2 Hz, 1H), 7.66 (dd, J = 11.5, 9.4 Hz, 2H), 7.42 (dd, J = 9.2, 1.9 Hz, 1H), 6.99 (t, J = 55.1 Hz, 1H), 6.82 (d, J = 9.6 Hz, 1H).

[0259] LC-MS (ESI): m / z 456 [M+H] + .

[0260] Preparation of Example 322 by General Procedure IIa (Method C): [ka]

[0261] Step A: 5-amino-6-bromo-4-chloro-2-(4-methoxybenzyl)pyridazin-3(2H)-one

[0262] To a solution of 5-amino-6-bromo-4-chloropyridazin-3(2H)-one (2.0 g, 8.91 mmol, 1.0 equiv) and K2CO3 (2.5 g, 17.8 mmol, 2.0 equiv) in DMF (20 mL) was added PMBCl (1.3 mL, 9.8 mmol, 1.1 equiv) and the reaction mixture was stirred at 80 °C for 14 h. After completion, the reaction mixture was diluted with water (50 mL). The mixture was diluted and extracted with EtOAc (50 mL × 3), the combined organic layers were washed with brine (50 mL), dried over Na2SO4, concentrated under reduced pressure, and the residue was purified by flash column chromatography on silica gel to give 5-amino-6-bromo-4-chloro-2-(4-methoxybenzyl)pyridine as a white solid. Dazin-3(2H)-one (2.0 g, 45%) was obtained. LC-MS (ESI): m / z 344 [M+H]+ .

[0263] Step B: Ethyl (E)-3-(4-amino-5-chloro-1-(4-methoxybenzyl)-6-oxo-1,6-dihydropyridazin-3-yl)acrylate

[0264] To a solution of 5-amino-6-bromo-4-chloro-2-(4-methoxybenzyl)pyridazin-3(2H)-one (2.0 g, 5.8 mmol, 1.0 equiv.), KCO (2.0 g, 14.5 mmol, 2.5 equiv.), and ethyl (E)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)acrylate (1.44 g, 6.4 mmol, 1.1 equiv.) in DMF (20 mL) was added Pd(dppf)Cl (0.43 g, 0.6 mmol, 0.1 equiv.), and the reaction mixture was stirred at 100 °C under a N atmosphere for 5 h. The reaction mixture was diluted with HO (30 mL), extracted with EtOAc (50 mL × 3), and the combined organic layers were washed with brine (40 mL), dried over NaSO, and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel to give ethyl (E)-3-(4-amino-5-chloro-1-(4-methoxybenzyl)-6-oxo-1,6-dihydropyridazine as a brown solid. 1.4 g (53%) of benzo-3-yl acrylate was obtained. LC-MS (ESI): m / z 364 [M+H] + .

[0265] Step C: 4-chloro-2-(4-methoxybenzyl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione

[0266] To a stirred solution of ethyl (E)-3-(4-amino-5-chloro-1-(4-methoxybenzyl)-6-oxo-1,6-dihydropyridazin-3-yl)acrylate (1.4 g, 3.85 mmol, 1.0 equiv) in EtOH (20 mL) was added KCO (1.6 g, 11.54 mmol, 3.0 equiv) and the reaction mixture was stirred at 80 °C for 14 h. The mixture was diluted with water (30 mL) and extracted with EtOAc (30 mL × 3). The combined organic layers were dried over NaSO and concentrated under reduced pressure to give crude 4-chloro-2-(4-methoxybenzyl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione (0.6 g, crude) as a brown solid, which was used in the next step without further purification. LC-MS (ESI): m / z 318 [M+H] + .

[0267] Step D: 4-(6-(difluoromethyl)pyridin-3-yl)-2-(4-methoxybenzyl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione

[0268] 4-Chloro-2-(4-methoxybenzyl)piperidin in 1,4-dioxane / H2O mixture (8 mL, 10 / 1, v / v) Lido[3,2-c]pyridazine-3,6(2H,5H)-dione (0.2 g, 0.63 mmol, 1.0 equiv.), 2-(4-(difluoromethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (152 mg, 0.88 mmol, 1.4 equiv.), KCO (217 mg, 1.57 mmol, 2.5 equiv.), and X-Phos (33 mg, 0.06 mmol, 0.1 equiv.) To a solution of 100 mg of Pd(OAc) (15 mg, 0.06 mmol, 0.1 equiv.) was added Pd(OAc) (15 mg, 0.06 mmol, 0.1 equiv.) and the reaction mixture was stirred at 110 °C under N atmosphere overnight. After completion, the reaction mixture was concentrated under reduced pressure and the residue was The compound was purified by column chromatography on a 600-ml column to give 4-(6-(difluoromethyl)pyridin-3-yl)-2-(4-methoxybenzyl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-diol as a white solid. The compound (120 mg, 46%) was obtained. LC-MS (ESI): m / z 411 [M+H] + .

[0269] Step E: 4-(6-(difluoromethyl)pyridin-3-yl)-2-(4-methoxybenzyl)-6-((2,2,2-trifluoroethyl)amino)pyrido[3,2-c]pyridazin-3(2H)-one

[0270] To a solution of 4-(6-(difluoromethyl)pyridin-3-yl)-2-(4-methoxybenzyl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione (100 mg, 0.24 mmol, 1.0 equiv) in DMF (2 mL) was added BOP (180 mg, 0.36 mmol, 1.5 equiv) and DIEA (157 mg, 1.22 mmol, 5.0 equiv), the reaction mixture was stirred at room temperature for 1 h, and then 2,2,2-trifluoroethan-1-amine (36 mg, 0.36 mmol, 1.5 equiv) was added. Amount) was added and the resulting mixture was stirred at room temperature for another 0.5 h. After completion, the reaction was quenched by adding ice water (10 mL), extracted with EtOAc (10 ml×3), dried over Na2SO4, concentrated under reduced pressure and the residue was purified by flash column chromatography on silica gel to give 4-(6-(difluoromethyl)pyridin-3-yl)-2-(4-methoxybenz ... (2,2,2-trifluoroethyl)amino)pyrido[3,2-c]pyridazin-3(2H)-one (80mg , 67%). LC-MS (ESI): m / z 492 [M+H] + .

[0271] Step F: 4-(6-(difluoromethyl)pyridin-3-yl)-6-((2,2,2-trifluoroethyl)amino)pyrido[3,2-c]pyridazin-3(2H)-one

[0272] To a solution of 4-(6-(difluoromethyl)pyridin-3-yl)-2-(4-methoxybenzyl)-6-((2,2,2-trifluoroethyl)amino)pyrido[3,2-c]pyridazin-3(2H)-one (80 mg, 0.16 mmol, 1.0 equiv.) in TFA (2 mL) was added TfOH (142 μL, 1.6 mmol, 10.0 equiv.) and EtSiH (128 μL, 0.8 mmol, 5.0 equiv.), and the reaction mixture was stirred at room temperature for 2 h. After completion, the reaction was quenched with 10 mL of NaHCO (saturated aqueous solution) and extracted with EtOAc (10 mL × 3). The combined organic layers were washed with brine (20 mL), dried over NaSO, concentrated under reduced pressure, and the residue was purified by flash filtration with silica gel. Purification by column chromatography gave 4-(6-(difluoromethyl)pyridinyl)- 3-(2,2,2-trifluoroethyl)amino)pyrido[3,2-c]pyridazin-3(2H)-o LC-MS (ESI): m / z 372 [M+H] + .

[0273] Step G: 4-(6-(difluoromethyl)pyridin-3-yl)-2-(4-(methoxy-d3)phenyl)-6-((2,2,2-trifluoroethyl)amino)pyrido[3,2-c]pyridazin-3(2H)-one (in operation) Example 322)

[0274] 4-(6-(difluoromethyl)pyridin-3-yl)-2-(4-(methoxy-d3)phenyl)-6-((2,2,2-trifluoroethyl)amino)pyrido[3,2-c]pyridazin-3(2H)-one (Example 322) 4-(6-(difluoromethyl)pyridin-3-yl)-6-((2,2,2-trifluoroethyl)amino)pyrido[3,2-c]pyridazin-3(2H)-one & 1-bromo- It was synthesized from 4-(methoxy-d3)benzene.

[0275] 1 H NMR (400 MHz, DMSO-d6) δ (ppm): 8.98 (s, 1H), 8.93 (t, J = 5.9 Hz, 1H), 8.30 (dd, J = 8.0 Hz, 1.6 Hz, 1H), 7.79 (d, J = 9.5 Hz, 1H), 7.73 (d, J = 8.2 Hz, 1H), 7.59-7.53 (m, 2H), 7.09-7.04 (m, 2H), 7.00 (t, J HF = 56.0 Hz, 1H), 6.93 (d, J = 9.5 Hz, 1H), 4.23-4.15 (m, 2H).

[0276] LC-MS (ESI): m / z 481 [M+H] + .

[0277] Using the procedure described above for General Procedure II (Method A), the appropriate starting materials may be used. The following compounds were synthesized using [Table 10-1] [Table 10-2] [Table 10-3] [Table 10-4] [Table 10-5] [Table 10-6] [Table 10-7]

[0278] Using the procedure described above for General Procedure II (Method B), the appropriate starting materials were used. The following compounds were synthesized using [Table 11-1] [Table 11-2] [Table 11-3]

[0279] Using the procedure described above for General Procedure II (Method C), the appropriate starting materials were used. The following compounds were synthesized using [Table 12-1] [Table 12-2] [Table 12-3]

[0280] General Procedure III: [ka]

[0281] Compounds of structure 3.6 were obtained through the scheme shown as General Procedure III. Starting with substituted pyridazinone 3.1 (synthesized by General Procedure I (Steps A-B)), the desired R group was introduced using Suzuki cross-coupling to generate vinyl compound 3.2. The double bond in 3.2 was then oxidized to give carbonyl 3.3. The desired R group was then added via Suzuki cross-coupling. The desired R5 group was introduced using a Horner-Wadsworth enzyme to give compound 3.4. The bicyclic heterocycle 3.5 was introduced using the Mons reaction and after tandem intramolecular cyclization Finally, the desired R1 group was introduced through an alkylation reaction to give compounds of structure 3.6.

[0282] Preparation of Example 194 by General Procedure III: [ka]

[0283] Step A: 5-amino-4-chloro-2-(2-methyl-2H-indazol-5-yl)-6-vinylpyridazin-3(2H)-one

[0284] A solution of 5-amino-6-bromo-4-chloro-2-(2-methyl-2H-indazol-5-yl)pyridazin-3(2H)-one (2 g, 5.67 mmol, 1.0 equiv., General Procedure I, (Steps A-B)), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (1.738 g, 11.34 mmol, 2.0 equiv.), Pd(dppf)Cl (0.826 g, 1.13 mmol, 0.2 equiv.), and KCO (2.336 g, 16.99 mmol, 3.0 equiv.) in dioxane / EtOH (48 mL, 3 / 1, v / v) was stirred at 80 °C for 16 h under a N atmosphere. The reaction mixture was filtered and concentrated under reduced pressure, and the residue was purified by flash column chromatography on silica gel to give 5-amino-4-chloro-2-(2-methyl-2H-indazol-5-yl)-6-vinylpyridazin-3(2H)-one (1.02 g, 60% yield) as a yellow solid. LC-MS (ESI): m / z 302 [M+H] + .

[0285] Step B: 4-amino-5-chloro-1-(2-methyl-2H-indazol-5-yl)-6-oxo-1,6- Dihydropyridazine-3-carbaldehyde

[0286] A solution of 5-amino-4-chloro-2-(2-methyl-2H-indazol-5-yl)-6-vinylpyridazin-3(2H)-one (1.6 g, 5.3 mmol, 1.0 equiv), NaIO (3.4 g, 15.9 mmol, 3.0 equiv), and KOsO-2HO (98 mg, 0.26 mmol, 0.05 equiv) in THF / HO (20 mL, 3:1) was stirred at room temperature for 3 h to give The resulting mixture was filtered and extracted with DCM (50 mL x 3). The filtrate was washed with brine (30 mL) and the combined The combined organic layers were dried over NaSO and concentrated under reduced pressure. The crude residue was purified by flash column chromatography on silica gel to give 4-amino-5-chloro-1-(2-methyl-2H-indazol-5-yl)-6-oxo-1,6-dihydropyridazine-3-carbaldehyde (1 g, 62% yield) as a white solid. LC-MS (ESI): m / z 304 [M+H] + .

[0287] Step C: 4-amino-5-(4-(difluoromethoxy)phenyl)-1-(2-methyl-2H-indazol-5-yl)-6-oxo-1,6-dihydropyridazine-3-carbaldehyde

[0288] 4-Amino-5-chloro-1-(2-methyl-2H-indazol-5-yl)-6-oxo-1,6-dihydropyridazine-3-carbaldehyde (650 mg, 2.15 mmol, 1.0 equiv.), 2-(4-(difluoromethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxa A solution of borolane (934 mg, 3.46 mmol, 1.6 equiv), Pd(OAc) (52 mg, 0.215 mmol, 0.1 equiv), X-Phos (205 mg, 0.43 mmol, 0.2 equiv), and KCO (594 mg, 4.3 mmol, 2.0 equiv) was stirred at 110 °C under a N atmosphere for 3 h. H2O was added, and the resulting mixture was extracted with EtOAc (30 mL x 3). The combined organic layers were dried over Na2SO4, concentrated under reduced pressure, and the residue was purified by flash column chromatography on silica gel to give 4-amino-5-(4-(difluoromethyl)-2-methyl-2-methyl-4-methyl ... (2-Methoxy)phenyl)-1-(2-methyl-2H-indazol-5-yl)-6-oxo-1,6-dihydropyridazine-3-carbaldehyde (650 mg, 69% yield). LC-MS (ESI): m / z 412 [M+H] + .

[0289] Step D: 4-(4-(difluoromethoxy)phenyl)-7-fluoro-2-(2-methyl-2H-indole) (2H,5H)-pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione

[0290] 4-Amino-5-(4-(difluoromethoxy)phenyl)-1-(2-methyl-2H-indazol-5-yl)-6-oxo-1,6-dihydropyridazine-3-carbaldehyde (150 mg, 0.36 mmol) in MeCN (5 mL) To a suspension of 2-(diethoxyphosphoryl)-2-fluoroethyl acetate (132 mg, 0.55 mmol, 1.5 equiv.) and ethyl 2-(diethoxyphosphoryl)-2-fluoroacetate (132 mg, 0.55 mmol, 1.5 equiv.), NaH (30% suspension in mineral oil, 175 mg, 2.19 mmol, 6.0 equiv.) was added in several portions at 0 °C under a N atmosphere. After stirring overnight at room temperature, the resulting mixture was cooled on ice. Pour into saturated NH4Cl (aqueous) (10 mL), then extract with EtOAc (10 mL x 3), and combine the organic layers The extract was washed with brine (20 mL), dried over NaSO, concentrated under reduced pressure, and the residue was purified by flash column chromatography on silica gel to give 4-(4-(difluoromethoxy)phenyl)-7-fluoro-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione (120 mg, 73% yield) as a yellow solid. LC-MS (ESI): m / z 454 [M+H] + .

[0291] 6-(2,2-Difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-7-fluoro-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 194) was synthesized from 4-(4-(difluoromethoxy)phenyl)-7-fluoro-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione and 2,2-difluoroethyl trifluoromethanesulfonate by a procedure similar to that described in General Procedure I (Step F).

[0292] 1 H NMR (400 MHz, DMSO-d6) δ: 8.56 (s, 1H), 8.10 (d, J = 10.0 Hz, 1H), 8.07 (d, J = 1.2 Hz, 1H), 7.91 (d, J = 9.2 Hz, 2H), 7.77 (d, J = 9.2 Hz, 1H), 7.51 (dd, J = 9.2 Hz, 2.0 Hz, 1H), 7.41 (t, J HF = 74.0 Hz, 1H), 7.32 (d, J = 9.2 Hz, 2H), 6.50 (tt, J HF = 54.0 Hz, 3.2 Hz, 1H), 4.71 (td, J HF = 15.2 Hz, J = 3.2 Hz, 2H), 4.28 (s, 3H).

[0293] LC-MS (ESI): m / z 518 [M+H] + .

[0294] Using the procedures described above for General Procedure III, using appropriate starting materials, The following compounds were synthesized: [Table 13]

[0295] Synthesis of 4-(4-bromophenyl)-6-ethoxy-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 198) [ka]

[0296] Step A: 4-(4-aminophenyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione

[0297] tert-Butyl (4-(2-(2-methyl-2H-indazol-5-yl)-3,6-indazol-5-yl)-3,6-indazol-5-yl) in TFA / DCM (1 mL / 5 mL) Dioxo-2,3,5,6-tetrahydropyrido[3,2-c]pyridazin-4-yl)phenyl)carbamate A solution of 4-chloro-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione and (4-((tert-butoxycarbonyl)amino)phenyl)boronic acid) (180 mg, 0.372 mmol) was stirred at room temperature for 1 hour. The reaction mixture was extracted with EtOAc (20 mL × 3). The combined organic layers were washed with brine (20 mL), dried over NaSO, and concentrated under reduced pressure, and the residue was purified by flash column chromatography on silica gel to give 4-(4-aminophenyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione (116 mg, 79% yield) as a yellow solid. Ta. LC-MS (ESI): m / z 385 [M+H] + .

[0298] Step B: 4-(4-bromophenyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione

[0299] To a solution of 4-(4-aminophenyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione (200 mg, 0.52 mmol, 1.0 equiv.) and CuBr (299 mg, 2.08 mmol, 4.0 equiv.) in ACN (10 mL), tert-butyl nitrite (215 mg, 2.08 mmol, 4.0 equiv.) was added dropwise over 10 min at 0 °C. The reaction mixture was stirred at room temperature for 4 h. The resulting mixture was quenched with NaSO (saturated aqueous solution) (20 mL) and extracted with EtOAc (10 mL × 3). The combined organic layers were washed with brine (20 mL), dried over NaSO, concentrated under reduced pressure, and the residue was purified by flash column chromatography on silica gel to give 4-(4-bromophenyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione (70 mg, 30% yield) as a yellow solid. LC-MS (ESI): m / z 448 [M+H] + .

[0300] Step C:

[0301] 4-(4-Bromophenyl)-6-ethoxy-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 198) was synthesized from 4-(4-bromophenyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione and iodoethane by general procedure I (method A, step F).

[0302] 1 H NMR (400 MHz, DMSO-d6) δ (ppm): 8.49 (s, 1H), 8.00 (d, J = 1.6 Hz, 1H), 7.97 (d, J = 9.6 Hz, 1H), 7.75 (d, J = 8.4 Hz, 2H), 7.70 (d, J = 9.2 Hz, 1H), 7.64 (d, J = 8.4 Hz, 2H), 7.45 (dd, J = 8.8 Hz, 1.6 Hz, 1H), 6.95 (d, J = 9.2 Hz, 1H), 4.33 (q, J = 7.2 Hz, 2H), 4.22 (s, 3H), 1.32 (t, J = 7.2 Hz, 3H).

[0303] LC-MS (ESI): m / z 476, 478 [M+H] + .

[0304] 4-(4-Bromophenyl)-6-isopropoxy-2-(2-methyl-2H-indazol-5-yl)pyridine Synthesis of do[3,2-c]pyridazin-3(2H)-one (Example 199) [ka]

[0305] 4-(4-Bromophenyl)-6-isopropoxy-2-(2-methyl-2H-indazol-5-yl)pyridine Do[3,2-c]pyridazin-3(2H)-one (Example 199) was synthesized from 4-(4-bromophenyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione and 2-iodopropane by general procedure I (method A, step F).

[0306] 1 H NMR (400 MHz, DMSO-d6) δ (ppm): 8.49 (s, 1H), 8.00 (dd, J = 2.0 Hz, 0.8 Hz, 1H), 7.96 (d, J = 9.4 Hz, 1H), 7.74-7.68 (m, 3H), 7.64 (d, J = 8.6 Hz, 2H), 7.45 (dd, J = 9.1 Hz, 2.0 Hz, 1H), 6.90 (d, J = 9.4 Hz, 1H), 5.15 (hept, J = 6.2 Hz, 1H), 4.22 (s, 3H), 1.30 (d, J = 6.2 Hz, 6H).

[0307] LC-MS (ESI): m / z 490, 492 [M+H] + .

[0308] Synthesis of 4-(4-bromophenyl)-6-(2,2-difluoroethoxy)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 200) [ka]

[0309] 4-(4-Bromophenyl)-6-(2,2-difluoroethoxy)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 200) was synthesized from 4-(4-bromophenyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione and 2,2-difluoroethyl trifluoromethanesulfonate by general procedure I (method A, step F).

[0310] 1 H NMR (400 MHz, DMSO-d6) δ (ppm): 8.51 (s, 1H), 8.07 (d, J = 9.2 Hz, 1H), 8.03 (d, J = 1.6 Hz, 1H), 7.76 (d, J = 8.8 Hz, 2H), 7.72 (d, J = 9.2 Hz, 1H), 7.66 (d, J = 8.4 Hz, 2H), 7.47 (dd, J = 9.2 Hz, 2.0 Hz, 1H), 7.08 (d, J = 9.2 Hz, 1H), 6.41 (tt, J HF = 54.4 Hz, 3.2 Hz, 1H), 4.59 (td, J HF = 15.2 Hz, J = 3.2 Hz, 2H), 4.23 (s, 3H).

[0311] LC-MS (ESI): m / z 512, 514 [M+H] + .

[0312] Synthesis of 2-(benzo[d]thiazol-6-yl)-4-(4-bromophenyl)-6-(2,2-difluoroethoxy)pyrido[3,2-c]pyridazin-3(2H)-one (Example 201) [ka]

[0313] A procedure similar to that described in Example 198 was followed: 2-(benzo[d]thiazol-6-yl)-4-(4-bromophenyl)-6-(2,2-difluoroethoxy)pyrido[3,2-c]pyridazine-3(2H)- tert-butyl(4-(2-benzo[d]))one (Example 201) was converted to tert-butyl(4-(2-benzo[d]))one by Steps A-B (Example 198). 2,2-dithiazol-6-yl)-3,6-dioxo-2,3,5,6-tetrahydropyrido[3,2-c]pyridazin-4-yl)phenyl)carbamate by general procedure I (Method A, Step F). Synthesized from fluoroethyl trifluoromethanesulfonate.

[0314] 1H NMR (400 MHz, DMSO-d6) δ (ppm): 9.53 (s, 1H), 8.54 (d, J = 2.1 Hz, 1H), 8.25 (d, J = 8.7 Hz, 1H), 8.07 (d, J = 9.4 Hz, 1H), 7.85 (dd, J = 8.7 Hz, 2.1 Hz, 1H), 7.76 (d, J = 8.6 Hz, 2H), 7.66 (d, J = 8.6 Hz, 2H), 7.09 (d, J = 9.4 Hz, 1H), 6.41 (tt, J HF = 54.4, 3.4 Hz, 1H), 4.60 (td, J = 15.2, 3.4 Hz, 2H).

[0315] LC-MS (ESI): m / z 515, 517 [M+H] + .

[0316] Synthesis of 2-(benzo[d]thiazol-6-yl)-4-(4-bromophenyl)-6-isopropoxypyrido[3,2-c]pyridazin-3(2H)-one (Example 202) [ka]

[0317] 2-(Benzo[d]thiazol-6-yl)-4-(4-bromophenyl)-6-isopropoxypyrido[3,2-c]pyridazin-3(2H)-one (Example 202) was prepared by general procedure I (method A, step F) to afford 2-(benz[d]thiazol-6-yl)-4-(4-bromophenyl)-6-isopropoxypyrido[3,2-c]pyridazin-3(2H)-one. It was synthesized from benzo[d]thiazol-6-yl)-4-(4-bromophenyl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione and 2-iodopropane.

[0318] 1 H NMR (400 MHz, DMSO-d6) δ 9.53 (s, 1H), 8.52 (d, J = 2.1 Hz, 1H), 8.24 (d, J = 8.7 Hz, 1H), 7.96 (d, J = 9.4 Hz, 1H), 7.83 (dd, J = 8.7, 2.1 Hz, 1H), 7.72 (d, J = 8.6 Hz, 2H), 7.65 (d, J = 8.6 Hz, 2H), 6.93 (d, J = 9.4 Hz, 1H), 5.16 (hept, J = 6.4 Hz, 1H), 1.31 (d, J = 6.4 Hz, 6H).

[0319] LC-MS (ESI): m / z 493, 495 [M+H] + .

[0320] 6-(2,2-Difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(3-(dimethylamino)-2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 203) & 4-(4-(difluoromethoxy)phenyl)-6-(dimethylamino)-2-(3-(dimethylamino) (2H)-2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 204) [ka]

[0321] Step A: 2-(3-bromo-2-methyl-2H-indazol-5-yl)-6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)pyrido[3,2-c]pyridazin-3(2H)-one

[0322] To a suspension of 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 125) (200 mg, 0.4 mmol, 1.0 equiv) in ACN (4 mL) was added NBS (75 mg, 0.42 mmol, 1.05 equiv). The reaction was then stirred at 80° C. for 3 h. The reaction was concentrated under reduced pressure and the residue was purified by fluo- ration on silica gel. Direct purification by flash column chromatography gave 2-(3-bromo-2-methyl-2H-indazol-5-yl)-6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)fluoro)-2-(4-bromo-2-methyl-2H-indazol-5-yl)-6-(2,2-difluoroethoxy)-4-(4-difluoromethoxy ... (phenyl)pyrido[3,2-c]pyridazin-3(2H)-one (160 mg, 69% yield). LC-MS (ESI): m / z 578; 580 [M+H] + .

[0323] Step B: 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(3-(dimethylamino)-2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one & 4-(4-(difluoromethoxy)phenyl)-6-(dimethylamino)-2-(3-(dimethylamino)-2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one

[0324] 2-(3-bromo-2-methyl-2H-indazol-5-yl)-6-(2,2-difluoromethyl) ... in toluene (3 mL) A suspension of (4-(difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)pyrido[3,2-c]pyridazin-3(2H)-one (160 mg, 0.28 mmol, 1.0 equiv.) was added to CsCO (90 mg, 0.28 mmol, 1.0 equiv.), Pd(dba) (25 mg, 0.028 mmol, 0.10 equiv.), Ru-Phos (26 mg, 0.055 mmol, 0.2 equiv.) and dimethylamine (2 M in THF) (1.4 mL, 2.8 mmol, 10.0 equiv.) were added. The reaction was then The mixture was sealed in a pressure tube and stirred under N2 atmosphere at 100 °C for 5 h. The reaction was cooled to room temperature, concentrated under reduced pressure, and the residue was purified by flash column chromatography on silica gel and RP preparative HPLC to give 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(3-(dimethylamino)-2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridyl. Dazin-3(2H)-one (Example 203) and 4-(4-(difluoromethoxy)phenyl)-6-(dimethylamino)-2-(3-(dimethylamino)-2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridin Dazin-3(2H)-one was obtained (Example 204).

[0325] Example 203:

[0326] 1 H NMR (400 MHz, DMSO-d6) δ (ppm): 8.11 (s, 1H), 8.08 (d, J = 9.4 Hz, 1H), 7.87 (d, J = 8.4 Hz, 2H), 7.55 (d, J = 9.2 Hz, 1H), 7.36 (d, J = 10.6 Hz, 1H), 7.35 (t, J HF = 74.0 Hz, 1H), 7.26 (d, J = 8.2 Hz, 2H), 7.06 (d, J = 9.3 Hz, 1H), 6.41 (tt, J HF = 54.4 Hz, J = 3.3 Hz, 1H), 4.59 (td, J HF = 15.1 Hz, J = 3.3 Hz, 2H), 3.99 (s, 3H), 2.97 (s, 6H).

[0327] LC-MS (ESI): m / z 543 [M+H] + .

[0328] Example 204:

[0329] 1 H NMR (400 MHz, DMSO-d6) δ (ppm): 8.04 (dd, J = 2.0, 0.8 Hz, 1H), 7.93-7.84 (m, 2H), 7.76 (d, J = 9.8 Hz, 1H), 7.52 (dd, J = 9.1, 0.8 Hz, 1H), 7.33 (dd, J = 9.2, 2.0 Hz, 1H), 7.31 (t, J HF = 74.2, 1H), 7.27 (d, J = 9.8 Hz, 1H), 7.22-7.16 (m, 2H), 3.99 (s, 3H), 3.23 (s, 3H), 3.17 (s, 3H), 2.96 (s, 6H).

[0330] LC-MS (ESI): m / z 506 [M+H] + .

[0331] 4-(4-(difluoromethoxy)phenyl)-2-(2-methyl-2H-indazol-5-yl)-6-proline Synthesis of pyripyrido[3,2-c]pyridazin-3(2H)-one (Example 205) [ka]

[0332] 6-chloro-4-(4-(difluoromethoxy)phenyl)-2-(2-methyl-2-methylpropional)-2-methylpropional in THF (5 mL) and NMP (1 mL) To a mixture of 4-chloro-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (synthesized by General Procedure I (Method C, steps E and G) from 4-chloro-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione and 2-(4-(difluoromethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane) (120 mg, 0.26 mmol, 1.0 equiv.) and Fe(acac) (93 mg, 0.26 mmol, 1.0 equiv.) was added n-propylmagnesium bromide (diethyl ether). To this mixture was added dropwise 4-(4-(difluoromethoxy)phenyl)-2-(2-methyl-2H-indazol-5-yl)-6-propylpyrido[3,2-c]pyridazin-3(2H)-one (Example 205). The crude mixture was extracted with EtOAc (10 mL × 3). The combined organic layers were washed with brine (20 mL), dried over NaSO, and concentrated under reduced pressure. The crude residue was purified by RP prep-HPLC to give 4-(4-(difluoromethoxy)phenyl)-2-(2-methyl-2H-indazol-5-yl)-6-propylpyrido[3,2-c]pyridazin-3(2H)-one (Example 205).

[0333] 1 H NMR (400 MHz, DMSO-d6) δ (ppm): 8.51 (s, 1H), 8.04 (s, 1H), 7.99 (d, J = 9.1 Hz, 1H), 7.83 (d, J = 8.6 Hz, 2H), 7.72 (d, J = 9.2 Hz, 1H), 7.49 (d, J = 9.1 Hz, 1H), 7.35 (t, J HF = 72.0 Hz, 1H), 7.29 (d, J = 9.3 Hz, 1H), 7.26 (d, J = 8.6 Hz, 2H), 4.23 (s, 3H), 2.78 (t, J = 7.4 Hz, 2H), 1.82-1.61 (m, 2H), 0.94 (t, J = 7.4 Hz, 3H).

[0334] LC-MS (ESI): m / z 462 [M+H] + .

[0335] Synthesis of 4-cyclohexyl-6-(2,2-difluoroethoxy)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 206) [ka]

[0336] Step A: 4-chloro-6-(2,2-difluoroethoxy)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one

[0337] 4,6-Dichloro-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c] in DMF (8 mL) Pyridazin-3(2H)-one (300 mg, 0.87 mmol, 1.0 equiv., synthesized by General Procedure I (Method A, Steps A-D)), 2,2-difluoroethyl trifluoromethanesulfonate (278 mg, 1.3 mmol) A mixture of CsCO (565 mg, 1.49 equiv.), and CsCO (565 mg, 1.7 mmol, 1.95 equiv.) was stirred at room temperature for 3 h and quenched with ice-water (10 mL). The crude mixture was extracted with EtOAc (10 mL × 3). The combined organic layers were washed with brine (20 mL), dried over NaSO, and concentrated under reduced pressure. The crude residue was purified by flash column chromatography on silica gel to give 4-chloro-6-(2,2-difluoroethoxy)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (250 mg, 74% yield) as a yellow solid. LC-MS (ESI): m / z 392 [M+H] + .

[0338] Step B: 4-Cyclohexyl-6-(2,2-difluoroethoxy)-2-(2-methyl-2H-indole) (2H)-pyrido[3,2-c]pyridazin-3(2H)-one

[0339] 4-Chloro-6-(2,2-difluoroethoxy)-2-(2-methyl-2H-indazol-5-yl)-2H,3H-pyrido[3,2-c]pyridazin-3-one (100 mg, 0.26 mmol, 1.0 equiv.), Fe(acac) (90 mg, 0.26 mmol, 1.0 equiv.), and a NMP / THF mixture (0.5 mL / 5 mL) were stirred at 0 °C under a N atmosphere. Cyclohexylmagnesium bromide (1 M in diethyl ether) (2.6 mL, 2.6 mmol, 10.0 (equivalent) was added dropwise at 0° C. The mixture was then warmed to room temperature and stirred overnight, the reaction was monitored by TLC, and upon completion, the reaction was quenched with ice water (10 mL) and the crude mixture was The crude residue was purified by RP prep-HPLC to give 4-cyclohexyl-6-(2,2-difluoroethoxy)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 206).

[0340] 1 H NMR (400 MHz, DMSO-d6) δ (ppm) 8.48 (s, 1H), 7.99-7.92 (m, 2H), 7.69 (d, J = 9.1 Hz, 1H), 7.39 (dd, J = 9.1, 2.0 Hz, 1H), 7.02 (d, J = 9.4 Hz, 1H), 6.50 (t, J HF = 54.2 Hz, 1H), 4.79 (td, J HF = 15.1, 3.2 Hz, 2H), 4.22 (s, 3H), 2.36-2.32 (m, 1H), 1.88-1.69 (m, 4H), 1.63-1.55 (m, 2H), 1.43-1.19 (m, 4H).

[0341] LC-MS (ESI): m / z 439 [M+H] + .

[0342] 6-(2,2-difluoroethoxy)-4-(4-hydroxyphenyl)-2-(2-methyl-2H-indazole Synthesis of (3,2-c)pyridazin-3(2H)-one (Example 207) [ka]

[0343] 4-chloro-6-(2,2-difluoroethoxy)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (100 mg, 0.25 mmol, 1.0 equiv.), Pd(OAc) (12 mg, 0.05 mmol, 0.2 equiv.), KCO (105 mg, 0.75 mmol, 3.0 equiv.) in a mixture of dioxane / water (5 mL, 10 / 1, v / v). A mixture of (4-hydroxyphenyl)boronic acid (53 mg, 0.38 mmol) and (4-hydroxyphenyl)boronic acid (53 mg, 0.38 mmol) was stirred at 100 °C under a N atmosphere. The reaction mixture was then poured into ice water (10 mL) and extracted with EtOAc (10 mL × 3). The combined organic layers were washed with brine (20 mL), dried over NaSO, and evaporated. Concentration under reduced pressure was carried out and the crude residue was purified by RP preparative HPLC to give 6-(2,2-difluoroethoxy)-4-(4-hydroxyphenyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 207).

[0344] 1H NMR (400 MHz, DMSO-d6) δ (ppm): 8.49 (s, 1H), 8.04-7.97 (m, 2H), 7.77-7.62 (m, 3H), 7.44 (dd, J = 9.1, 1.9 Hz, 1H), 7.03 (d, J = 9.4 Hz, 1H), 6.83 (d, J = 8.4 Hz, 2H), 6.42 (tt, J HF = 54.5, 3.5 Hz, 1H), 4.61 (td, J = 15.0, 3.5 Hz, 2H), 4.22 (s, 3H).

[0345] LC-MS (ESI): m / z 450 [M+H] + .

[0346] Synthesis of 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(3-ethyl-2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 208) ) [ka]

[0347] Step A: 1-(5-bromo-2-methyl-2H-indazol-3-yl)ethanol

[0348] To a solution of 5-bromo-2-methyl-2H-indazole (500 mg, 2.37 mmol, 1 equiv.) in THF (10 mL) was added n-BuLi (2.5 M in hexanes) (4.7 mL, 11.85 mmol, 5 equiv.) dropwise at −65° C. under a N atmosphere. After 3 h, acetaldehyde (5 M in THF) (0.6 mL, 3.0 mmol, 1.27 equiv.) was added. The reaction was then gradually warmed to room temperature and stirred for 16 h. The reaction mixture was then poured into saturated aqueous NH4Cl (10 mL) and extracted with EtOAc (10 mL × 3). The combined organic layers were concentrated to give a salt solution. The crude residue was purified by flash chromatography on silica gel to give 1-(5-bromo-2-methyl-2H-indazol-3-yl)ethanol (500 mg, 83% yield) as a yellow solid. LC-MS (ESI): m / z 255, 257 [M+H] + .

[0349] Step B: 5-Bromo-3-ethyl-2-methyl-2H-indazole

[0350] 1-(5-bromo-2-methyl-2H-indazol-3-yl)ethanol (200 mg, 0.78 mmol, 1 equiv.), triethylsilane (453 mg, 3.9 mmol, 5 equiv.), TFA (889 mg, 7.8 mmol, 10 equiv.) in DCM (5 mL). The mixture was stirred at 40° C. for 16 hours. Then, the mixture was diluted with saturated aqueous NaHCO3 (10 mL) The combined organic layer was dried over Na2SO4 and evaporated under reduced pressure. The crude residue was purified by flash chromatography on silica gel to give 5-bromo-3-ethyl-2-methyl-2H-indazole (80 mg, 43% yield) as a yellow solid. LC-MS (ESI): m / z 239, 241 [M+H] + .

[0351] 6-(2,2-Difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(3-ethyl-2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 208) was prepared manually. 6-(2,2-difluoroethoxy)-4-(4-(difluoromethacrylate)-2-methyl-2-methyl-4 ... It was synthesized from (oxy)phenyl)pyrido[3,2-c]pyridazin-3(2H)-one and 5-bromo-3-ethyl-2-methyl-2H-indazole.

[0352] 1 H NMR (400 MHz, DMSO-d6) δ: 8.13 (d, J = 9.2 Hz, 1H), 8.07 (dd, J = 2.0 Hz, 0.8 Hz, 1H), 7.93 (d, J = 8.8 Hz, 2H), 7.68 (dd, J = 9.2 Hz, 0.8 Hz, 1H), 7.48 (dd, J = 8.8 Hz, 1.6 Hz, 1H), 7.41 (t, J HF = 74.0 Hz, 1H), 7.31 (d, J = 8.8 Hz, 2H), 7.13 (d, J = 9.2 Hz, 1H), 6.47 (tt, J HF = 54.4 Hz, 3.2 Hz, 1H), 4.65 (td, J HF = 15.2 Hz, 3.2 Hz, 2H), 4.20 (s, 3H), 3.17 (q, J = 7.2 Hz, 2H), 1.33 (t, J = 7.2 Hz, 3H).

[0353] LC-MS (ESI): m / z 528 [M+H] + .

[0354] 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-8-(hydroxymethyl) (2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one ( Example 209) and synthesis of 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-8-(difluoromethyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 210) [ka]

[0355] Step A: 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-8-( Hydroxymethyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one

[0356] 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(2,2-difluoroethoxy)phenyl)-2 ... (2-methyl-2H-indazol-5-yl)-8(((tetrahydro-2H-pyran-2-yl)oxy)methyl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 142, General Procedure I (Method A, Step To a solution of (E)-methyl 4-(tetrahydro-2H-pyran-2-yloxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)but-2-enoate (synthesized from Tetrahedron 2012, 68, 3444-3449) (180 mg, 0.3 mmol, 1.0 equiv.) according to Steps E-F, 1N HCl (aq.) (1 mL, 1 mmol, 3.3 equiv.) was added at room temperature. The resulting mixture was stirred at room temperature for 2 hours. The reaction was then quenched with ice water (20 mL) and extracted with EtOAc (20 mL × 3). The combined organic layers were washed with brine (20 mL), dried over NaSO, concentrated under reduced pressure, and the residue was purified by flash column chromatography on silica gel to give 6-(2,2-difluoroethoxy)-2-methyl-2-propanol. hydroxy)-4-(4-(difluoromethoxy)phenyl)-8-(hydroxymethyl)-2-(2-methyl-2H-isothiazolinone To give (indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 209).

[0357] 1H NMR (400 MHz, DMSO-d6) δ (ppm): 8.50 (s, 1H), 8.04 (s, 1H), 7.84 (d, J = 8.4 Hz, 2H), 7.70 (d, J = 9.6 Hz, 1H), 7.49 (d, J = 8.8 Hz, 1H), 7.34 (t, J HF = 74.0 Hz, 1H), 7.24 (d, J = 8.4 Hz, 2H), 6.94 (s, 1H), 6.39 (tt, J HF = 54.4 Hz, 3.2 Hz, 1H), 5.63 (s, 1H), 4.83 (s, 2H), 4.58 (td, J HF = 15.2 Hz, 3.2 Hz, 2H), 4.22 (s, 3H).

[0358] LC-MS (ESI): m / z 530 [M+H] + .

[0359] Step B: 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(2-methyl-2H-indazol-5-yl)-3-oxo-2,3-dihydropyrido[3,2-c]pyridazine-8- Carbaldehyde

[0360] To a solution of 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-8-(hydroxymethyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (60 mg, 0.1 mmol, 1.0 equiv.) in CHCl3 (5 mL) was added freshly activated MnO2 (96 mg, 1.0 mmol, 10.0 equiv.) in one portion. The resulting mixture was then stirred at room temperature for 16 h. The reaction progress was monitored by TLC, and upon completion, the reaction mixture was passed through a short pad of Celite®. The mixture was filtered, and the filtrate was concentrated under reduced pressure to give crude 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(2-methyl-2H-indazol-5-yl)-3-oxo-2,3-dihydropyrido[3,2-c]pyridazine-8-carbaldehyde (50 mg) as a yellow solid, which was further purified. It was used in the next step without further purification.

[0361] LC-MS: m / z 528 [M+H] + .

[0362] Step C: 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-8-( Difluoromethyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one

[0363] 6-(2,2-Difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(2-methyl-2H-indazol-5-yl)-3-oxo-2,3-dihydropyrido[3,2-c]pyridazine-8- in DCM (3 mL) To a solution of carbaldehyde (50 mg, crude) was added DAST (46 mg) at −60° C. After addition, the mixture was cooled to room temperature. The mixture was allowed to warm to rt and stirred for another 16 h. After the reaction was completed, ice water (10 mL) was added and extracted with EtOAc (10 mL × 3). The combined organic layer was washed with brine (20 mL), dried over NaSO, and concentrated under reduced pressure. The residue was purified by RP prep-HPLC to give 6-(2,2-difluoroethoxy)-4-(4-(difluoroethoxy)- ... Fluoromethoxy)phenyl)-8-(difluoromethyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 210).

[0364] 1H NMR (400 MHz, DMSO-d6) δ (ppm): 8.53 (s, 1H), 8.08 (d, J = 2.0 Hz, 1H), 7.86 (d, J = 8.8 Hz, 2H), 7.73 (d, J = 9.2 Hz, 1H), 7.51 (dd, J = 9.2 Hz, 2.0 Hz, 1H), 7.45 (t, J HF = 53.6 Hz, 1H), 7.36 (t, J HF = 74.0 Hz, 1H), 7.30-7.25 (m, 3H), 6.41 (tt, J HF = 54.4 Hz, 3.2 Hz, 1H), 4.61 (td, J HF = 15.2 Hz, 3.2 Hz, 2H), 4.23 (s, 3H).

[0365] LC-MS (ESI): m / z 550 [M+H] + .

[0366] 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-8((dimethylamine) Synthesis of (2-methyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 211) [ka]

[0367] Step A: (6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(2-methyl-2H-indazol-5-yl)-3-oxo-2,3-dihydropyrido[3,2-c]pyridazin-8-yl)methyl methanesulfonate

[0368] 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-8-( To a solution of (hydroxymethyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (90 mg, 0.17 mmol, 1.0 equiv., Example 209), MsCl (29 mg, 0.25 mmol, 1.47 equiv.) and TEA (34 mg, 0.34 mmol, 2.0 equiv.) were added at room temperature. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was then poured into ice water (10 mL) and extracted with EtOAc (10 mL x 3). The combined organic layers were washed with brine (20 mL), dried over NaSO, concentrated under reduced pressure, and the crude residue was purified by flash column chromatography on silica gel to give (6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(2-methyl-2H-indazol-5-yl)-3-oxo-2,3-dihydropyrido[3,2-c]pyridazin-8-yl) as a yellow solid. Methyl methanesulfonate (90 mg, 87% yield) was obtained. LC-MS: m / z 608 [M+H] + .

[0369] Step B: 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-8-((dimethylamino)methyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazine On-3(2H)-On

[0370] To a solution of (6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(2-methyl-2H-indazol-5-yl)-3-oxo-2,3-dihydropyrido[3,2-c]pyridazin-8-yl)methyl methanesulfonate (50 mg, 0.08 mmol, 1.0 equiv.) in DMSO (3 mL), NH(Me)HCl salt (33 mg, 0.41 mmol, 5.1 equiv.), NaI (23 mg, 0.16 mmol, 2.0 equiv.), and NaHCO (14 mg, 0.16 mmol, 2.0 equiv.) were added at room temperature. The reaction mixture was stirred at 80 °C for 2 h. The reaction mixture was then poured into ice water (10 mL) and extracted with EtOAc (10 mL × 3). The combined organic layers were washed with brine (20 mL), dried over NaSO, and concentrated under reduced pressure. The crude residue was purified by RP preparative HPLC to give 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-8((difluoroethoxy)phenyl). Methylamino)methyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one was obtained (Example 211).

[0371] 1 H NMR (400 MHz, DMSO-d6) δ (ppm) 8.52 (s, 1H), 8.05 (d, J = 2.0 Hz, 1H), 7.84 (d, J = 8.8 Hz, 2H), 7.72 (d, J = 9.2 Hz, 1H), 7.49 (dd, J = 9.2, 2.0 Hz, 1H), 7.36 (t, J HF = 74.0 Hz, 1H), 7.25 (d, J = 8.8 Hz, 2H), 6.96 (s, 1H), 6.40 (tt, J HF = 54.4, 3.4 Hz, 1H), 4.23 (s, 3H), 3.77 (s, 2H), 2.29 (s, 6H).

[0372] LC-MS (ESI): m / z 557 [M+H] + .

[0373] Synthesis of 8-amino-6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 212) [ka]

[0374] Step A: 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(2- Methyl-2H-indazol-5-yl)-3-oxo-2,3-dihydropyrido[3,2-c]pyridazine-8-carboxylic acid

[0375] To a mixture of 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(2-methyl-2H-indazol-5-yl)-3-oxo-2,3-dihydropyrido[3,2-c]pyridazine-8-carbaldehyde (120 mg, 0.23 mmol, 1.0 equiv., synthesized as in Example 210, Step B) and HCOH (0.1 mL, 2.65 mmol, 11.5 equiv.) in HO (0.5 mL) was added HO (30 wt.% in HO, 0.1 mL, 1.14 mmol, 5 equiv.) slowly at 4 °C. The mixture was stirred at room temperature for 6 hours. The mixture was poured into H2O (10 mL) and extracted with ETOAC (10 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, and concentrated under reduced pressure to give a crude residue, which was purified by fluo- ration on silica gel. Purification by column chromatography gave (6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(2-methyl-2H-indazol-5-yl)-3-oxo-2,3-dihydropyrido[3,2-c]pyridazine-8-carboxylic acid (80 mg, 52% yield) as a yellow solid. LC-MS (ESI): m / z 544 [M+H] + .

[0376] Step B: 8-amino-6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)- (2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one

[0377] 6-(2,2-Difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(2-methyl-2H-indazol-5-yl)-3-oxo-2,3-dihydropyrido[3,2-c]pyridazine-8- in DMF (2 mL) To a solution of carboxylic acid (80 mg, 0.15 mmol, 1.5 equiv.) was added TEA (22 mg, 0.22 mmol, 1.45 equiv.) and DPPA (61 mg, 0.22 mmol, 1.45 equiv.). The mixture was stirred at room temperature for 3 hours. Then, 0.3 mL of water was added to the solution, and the reaction mixture was stirred at 100° C. overnight. The reaction was cooled to room temperature. The mixture was cooled, poured into HO (10 mL), and extracted with EtOAc (10 mL × 3). The combined organic layers were washed with brine (20 mL), dried over NaSO, concentrated under reduced pressure, and the crude residue was purified by RP prep-TLC to give 8-amino-6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 212). .

[0378] 1 H NMR (400 MHz, DMSO-d6) δ (ppm): 8.49 (s, 1H), 8.05 (d, J = 1.2 Hz, 1H), 7.83 (d, J = 8.8 Hz, 2H), 7.69 (d, J = 9.2 Hz, 1H), 7.51 (dd, J = 9.2 Hz, 1.6 Hz, 1H), 7.32 (t, J HF = 74.0 Hz, 1H), 7.23-7.14 (m, 4H), 6.31 (tt, J HF= 54.0 Hz, 3.6 Hz, 1H), 5.83 (s, 1H), 4.47 (td, J = 14.8 Hz, 3.6 Hz, 2H), 4.22 (s, 3H).

[0379] LC-MS (ESI): m / z 515 [M+H] + .

[0380] 6-(2,2-difluoroethoxy)-2-(2-methyl-2H-indazol-5-yl)-4-((trimethyl Silyl)ethynyl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 213), 6-(2,2-difluoroethoxy)-4-ethynyl-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin Indazol-3(2H)-one (Example 214) & 4-((1H-pyrazol-3-yl)ethynyl)-6-(2,2-difluoroethoxy)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one Synthesis of (Example 215) [ka]

[0381] Step A: 6-(2,2-difluoroethoxy)-2-(2-methyl-2H-indazol-5-yl)-4-((trimethylsilyl)ethynyl)pyrido[3,2-c]pyridazin-3(2H)-one

[0382] 4-chloro-6-(2,2-difluoroethoxy)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (180 mg, 0.46 mmol, 1.0 equiv., performed) in dioxane (5 mL) A mixture of Pd(PPh) (synthesized as in Example 206, Step A), trimethyl((tributylstannyl)ethynyl)silane (268 mg, 0.69 mmol, 1.5 equiv.), and Pd(PPh) (58 mg, 0.05 mmol, 0.11 equiv.) was stirred at 80 °C for 15 h under a N atmosphere. The reaction mixture was concentrated under reduced pressure, and the crude residue was purified by flash column chromatography on silica gel to give 6-(2,2-difluoroethoxy)-2-(2-methyl-2H-indazol-5-yl)-4-((trimethylsilyl)ethynyl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 213).

[0383] 1 H NMR (400 MHz, DMSO-d6) δ (ppm): 8.50 (s, 1H), 8.08 (d, J = 9.6 Hz, 1H), 7.98 (d, J = 1.6 Hz, 1H), 7.70 (d, J = 9.2 Hz, 1H), 7.41 (dd, J = 9.2 Hz, 2.0 Hz, 1H), 7.09 (d, J = 9.2 Hz, 1H), 6.54 (tt, J HF = 54.8 Hz, 3.6 Hz, 1H), 4.82 (td, J HF = 14.4 Hz, 3.6 Hz, 2H), 4.22 (s, 3H), 0.28 (s, 9H).

[0384] LC-MS (ESI): m / z 454 [M+H] + .

[0385] Step B: 6-(2,2-difluoroethoxy)-4-ethynyl-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one

[0386] A mixture of 6-(2,2-difluoroethoxy)-2-(2-methyl-2H-indazol-5-yl)-4-((trimethylsilyl)ethynyl)pyrido[3,2-c]pyridazin-3(2H)-one (126 mg, 0.28 mmol, 1.0 equiv.) and CsCO (183 mg, 0.56 mmol, 2.0 equiv.) in 2,2-difluoroethan-1-ol (4 mL). The mixture was stirred at room temperature for 4 hours. The resulting mixture was poured into H2O (10 mL) and extracted with EtOAc (10 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, and concentrated under reduced pressure. The crude residue was purified by flash column chromatography on silica gel to give 6-(2,2-difluoroethoxy)-4-ethynyl-2-(2-methyl-2H-indazol-5-yl) Pyrido[3,2-c]pyridazin-3(2H)-one was obtained (Example 214).

[0387] 1 H NMR (400 MHz, DMSO-d6) δ (ppm): 8.51 (s, 1H), 8.08 (d, J = 9.2 Hz, 1H), 7.99 (d, J = 1.6 Hz, 1H), 7.72 (d, J = 9.6 Hz, 1H), 7.41 (dd, J = 9.2 Hz, 2.0 Hz, 1H), 7.10 (d, J = 9.2 Hz, 1H), 6.52 (tt, J HF = 54.4 Hz, 3.2 Hz, 1H), 5.02 (s, 1H), 4.83 (td, J HF = 14.8 Hz, 3.2 Hz, 2H), 4.23 (s, 3H).

[0388] LC-MS (ESI): m / z 382 [M+H] + .

[0389] Step C: 4-((1H-pyrazol-3-yl)ethynyl)-6-(2,2-difluoroethoxy)-2-(2- Methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one

[0390] 6-(2,2-Difluoroethoxy)-4-ethynyl-2-(2-methyl-2H-indazole) in DMF (5 mL) (Iron-5-yl)pyrido[3,2-c]pyridazin-3(2,2H)-one (50 mg, 0.13 mmol, 1.0 equiv.), DIPEA (50 mg, 0.39 mmol, 3.0 equiv.), Pd(PPh3)2Cl2 (10 mg, 0.013 mmol, 0.1 equiv.), and 3-iodo-1H-pyrazole (101 mg, 0.52 mmol, 4.0 equiv.) and CuI A mixture of (25 mg, 0.13 mmol, 1.0 equiv) was stirred at room temperature under N2 atmosphere for 4 h. The reaction mixture was then poured into H2O (10 mL) and extracted with EtOAc (10 mL × 3). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, and concentrated under reduced pressure. The crude residue was purified by RP prep-HPLC to give 4-((1H-pyridinyl)- ... To give)-2-(2-methyl-2H-indazol-3-yl)ethynyl)-6-(2,2-difluoroethoxy)-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (example 215).

[0391] 1 H NMR (400 MHz, DMSO-d6) δ (ppm): 13.39 (br, s, 1H), 8.52 (s, 1H), 8.09 (d, J = 9.2 Hz, 1H), 8.01 (s, 1H), 7.91-7.81 (m, 1H), 7.72 (d, J = 9.2 Hz, 1H), 7.44 (dd, J = 9.2 Hz, 2.0 Hz, 1H), 7.11 (d, J = 9.2Hz, 1H), 6.62 (s, 1H), 6.55 (tt, J HF = 54.4 Hz, 3.2 Hz, 1H), 4.88 (td, J HF = 14.8 Hz, 3.2 Hz, 2H), 4.23 (s, 3H).

[0392] LC-MS (ESI): m / z 448 [M+H] + .

[0393] 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(4-oxocyclohexane Synthesis of (2H)pyrido[3,2-c]pyridazin-3(2H)-one (Example 216) [ka]

[0394] 6-(2,2-Difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)pyrido[3,2-c]pyridazin-3(2H)-one (6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)pyrido[3,2-c]pyridazin-3(2H)-one) was prepared by General Procedure II (Method A, Step G) in THF (1.5 mL). (trimethoxy)phenyl)pyrido[3,2-c]pyridazin-3(2H)-one & 1,4-dioxaspiro[4.5] A solution of (synthesized from dec-7-en-8-yl trifluoromethanesulfonate) (10 mg, 0.0197 mmol) and concentrated HCl (0.1 mL) was stirred at room temperature for 2 h. The reaction was poured into saturated aqueous NaHCO3 (5 mL) and extracted with EA (5 mL × 3). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, and concentrated under reduced pressure. Then, a mixture of the residue and Rh(PPh3)3Cl (18 mg, 0.0197 mmol) in toluene (3 mL) was stirred at room temperature overnight under an H2 atmosphere. The mixture was then diluted with H2O (5 mL) and extracted with ETOAC (5 mL × 3). The combined organic layers were washed with brine (10 mL), dried over NaSO, and concentrated under reduced pressure. The crude residue was purified by RP prep-HPLC to give 6-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-2-(4-oxocyclohexane). Xyl)pyrido[3,2-c]pyridazin-3(2H)-one was obtained (Example 216).

[0395] 1 H NMR (400 MHz, DMSO-d6) δ (ppm): 8.02 (d, J = 9.6 Hz, 1H), 7.82 (d, J = 8.8 Hz, 2H), 7.34 (t, J HF = 74.0 Hz, 1H), 7.24 (d, J = 8.8 Hz, 2H), 7.02 (d, J = 9.6 Hz, 1H), 6.37 (tt, J HF = 54.4 Hz, 2.8 Hz, 1H), 5.62 (hept, J = 4.8 Hz, 1H), 4.54 (td, J HF = 14.8 Hz, 2.8 Hz, 2H), 2.78-2.63 (m, 2H). 2.45-2.23 (m, 2H), 2.29-2.14 (m, 1H) (HCO2H salt).

[0396] LC-MS (ESI): m / z 466 [M+H] + .

[0397] Synthesis of 4-(4-(difluoromethoxy)phenyl)-6-ethoxy-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 217) [ka]

[0398] 6-chloro-4-(4-(difluoromethoxy)phenyl-2-(2-methyl-2H-yne)-2-yl)propanol in EtOH (8 mL) 4-chloro-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (4-chloro-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one by General Procedure I (Method C, Steps E, G) A mixture of 4-(4-(difluoromethoxy)phenyl)-3,6(2H,5H)-dione & 2-(4-(difluoromethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (120 mg, 0.26 mmol, 1.0 equiv.) and EtONa (177 mg, 2.6 mmol, 10.0 equiv.) was stirred at 40 °C for 3 h. The reaction mixture was then poured into ice water (10 mL) and extracted with DCM (10 mL × 3). The combined organic layers were dried over Na2SO4 and concentrated under reduced pressure. The crude residue was purified by RP prep-HPLC to give 4-(4-(difluoromethoxy)phenyl)-3,6(2H,5H)-dione & 2-(4-(difluoromethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (120 mg, 0.26 mmol, 1.0 equiv.) and EtONa (177 mg, 2.6 mmol, 10.0 equiv.). The reaction mixture was then poured into ice water (10 mL) and extracted with DCM (10 mL × 3). The combined organic layers were dried over Na2SO4 and concentrated under reduced pressure. The crude residue was purified by RP prep-HPLC to give 4-(4-(difluoromethoxy)phenyl)-3,6(2H,5H)-dione & 2-(4-(difluoromethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (120 mg, 0.26 mmol, 1.0 equiv.). 2-(2-methyl-2H-indazol-5-yl)-6-ethoxy-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazin-3(2H)-one (Example 217).

[0399] 1 H NMR (400 MHz, DMSO-d6) δ: 8.49 (s, 1H), 8.04 (d, J = 1.2 Hz, 1H), 7.97 (d, J = 9.2 Hz, 1H), 7.86 (d, J = 8.8 Hz, 2H), 7.70 (d, J = 9.2 Hz, 1H), 7.45 (dd, J = 9.2 Hz, 2.0 Hz, 1H), 7.34 (t, J HF = 74.0, 1H), 7.24 (d, J = 8.8 Hz, 2H), 6.95 (d, J = 9.6 Hz, 1H), 4.34 (q, J = 7.2 Hz, 2H), 4.22 (s, 3H), 1.32 (t, J = 7.2 Hz, 3H).

[0400] LC-MS (ESI): m / z 464 [M+H] + .

[0401] Synthesis of 5-(6-(2,2-difluoroethoxy)-2-(2-methyl-2H-indazol-5-yl)-3-oxo-2,3-dihydropyrido[3,2-c]pyridazin-4-yl)-1H-indole-3-carbonitrile (Example 331) [ka]

[0402] 7-(2,2-Difluoroethoxy)-3-(2-methyl-2H-indazol-5-yl)-1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)-3,4-dihydro in DCM (3 mL) Pyrido[2,3-d]pyrimidin-2(1H)-one (54 mg, 0.09 mmol, 1.0 equiv.) (4-chloro-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyrimidin-2(1H)-one) (Method A, Steps E & F) Pyridazine-3,6(2H,5H)-dione & 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2- To a solution of (2-(trimethylsilyl)ethoxy)methyl)-1H-indole-3-carbonitrile (synthesized from reference: WO2018215316)) was added TFA (1 mL), and the reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was then concentrated under reduced pressure, and the residue was redissolved in MeOH (2 mL) and concentrated NH4OH (1 mL), and the resulting mixture was stirred at room temperature overnight. Upon completion, the reaction mixture was concentrated under reduced pressure. The mixture was concentrated under reduced pressure and the residue was purified by RP preparative HPLC to give 5-(6-(2,2-difluoroethoxy)-2-(2- Methyl-2H-indazol-5-yl)-3-oxo-2,3-dihydropyrido[3,2-c]pyridazin-4-yl)-1H-indole-3-carbonitrile was obtained (Example 331).

[0403] 1 H NMR (400 MHz, DMSO-d6) δ (ppm): δ 12.29 (s, 1H), 8.50 (s, 1H), 8.29 (s, 1H), 8.12 (d, J = 0.9 Hz, 1H), 8.10-8.01 (m, 2H), 7.75 (dd, J = 8.6 Hz, 1.5 Hz, 1H), 7.71 (d, J = 9.2 Hz, 1H), 7.60 (d, J = 8.7 Hz, 1H), 7.49 (dd, J = 9.1 Hz, 2.0 Hz, 1H), 7.06 (d, J = 9.4 Hz, 1H), 6.38 (tt, J HF = 54.5 Hz, J = 3.3 Hz, 1H), 4.54 (td, J HF = 15.0 Hz, J = 3.4 Hz, 2H), 4.22 (s, 3H).

[0404] LC-MS (ESI): m / z 498 [M+H] + .

[0405] 5-(6-(ethylamino)-2-(2-methyl-2H-indazol-5-yl)-3-oxo-2,3-dihydro Synthesis of pyrido[3,2-c]pyridazin-4-yl)-1H-indole-3-carbonitrile (Example 332) [ka]

[0406] 5-(6-(ethylamino)-2-(2-methyl-2H-indazol-5-yl)-3-oxo-2,3-dihydropyrido[3,2-c]pyridazin-4-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indole-3-carbonitrile (100 mg, 0.17 mmol) (4-chloro-2-(2-methyl-2H-indazol-5-yl)pyrido[3,2-c]pyridazine-3,6(2H,5H)-dione & 5-(4,4,5,5-tetramethyl-1,3,2-dioxabonitrile) according to General Procedure I (Method A, Step E; Method D, Step I) To a solution of (loran-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indole-3-carbonitrile (synthesized from WO2018215316)), TFA (6 mL) was added, and the reaction mixture was stirred at room temperature for 12 hours. The reaction mixture was then concentrated under reduced pressure, and the residue was dissolved in MeOH (4 mL) and concentrated The concentrate was redissolved in NH4OH (2 mL) and the resulting mixture was stirred at room temperature for an additional 6 h. The reaction mixture was concentrated under reduced pressure, and the residue was purified by RP preparative HPLC to give 5-(6-(ethylamino)-2-(2-methyl-2H-indazol-5-yl)-3-oxo-2,3-dihydropyrido[3,2-c]pyridazin-4-yl)-1H-indole-3-carbonitrile (Example 332).

[0407] 1 H NMR (400 MHz, DMSO-d6) (ppm): δ 12.17 (s, 1H), 8.45 (s, 1H), 8.27 (s, 1H), 8.22 (s, 1H), 8.16 (s, 1H), 7.96 (s, 1H), 7.76 (d, J = 9.0 Hz, 1H), 7.66 (d, J = 9.1 Hz, 1H), 7.64 (d, J = 9.1 Hz, 1H), 7.51 (d, J = 8.9 Hz, 1H), 7.43 (d, J = 8.9 Hz, 1H), 6.78 (d, J = 9.3 Hz, 1H), 4.21 (s, 3H), 3.31 (q, J = 7.1 Hz, 2H), 1.14 (t, J = 7.1Hz, 3H).

[0408] LC-MS (ESI): m / z 461 [M+H] + .

[0409] Biochemical assays

[0410] Mat2A protein was expressed by recombinant baculovirus in SF9 infected cells using the Bac-to-Bac system and transfected into the pFASTBAC1 vector (Invitrogen, CA, USA). The recombinant MAT2A was cloned into a chromatin containing 100 kDa of 100 kJ / ml (Hollsbad). Recombinant MAT2A homodimers were isolated from cell lysates of 150 g of infected cells using column chromatography. Recombinant MAT2A homodimers were eluted with 250 and 500 mM imidazole, and MAT2A-containing fractions were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and pooled.

[0411] To determine the inhibitory activity of compounds against MAT2A homodimers, protein was diluted to 4 μg / mL in assay buffer (50 mM Tris, pH 8.0, 50 mM KCl, 15 mM MgCl, 0.3 mM EDTA, 0.005% [w / v] bovine serum albumin [BSA]). Test compounds were prepared in 100% dimethyl sulfoxide (DMSO) at 50× the desired final concentration. 1 μL aliquots of compound dilutions were added to 40 μL of enzyme. The enzyme assay was performed using 10 μL of the substrate mixture. The reaction mixture was incubated with 1x assay buffer (500 μM ATP, pH 7.0, 400 μM L-methionine). The mixture was then incubated at 25°C for a further 60 minutes. The reaction was stopped and the free phosphate released from the theoretical amount of enzyme by the production of S-adenosylmethionine (SAM) was measured using a PiColorLock Gold kit (Innova Biosciences, UK). The absolute production was The concentration was determined by comparison with a standard curve in potassium phosphate buffer, pH 8.0.

[0412] Certain compounds disclosed herein were tested in the aforementioned assays and demonstrated to inhibit MAT2A The inhibitory effect is evaluated according to the following score: IC 50 was determined as shown in Table 2 below: (A) <100 nM (>40% maximal inhibition), (B) 100 nM to 1 μM (>40% maximal inhibition), and (C) 1 μM to 10 μM (>40% maximal inhibition).

[0413] Cellular assay for target binding (SAM)

[0414] Measurement of MAT2A activity in cells was achieved by direct quantitation of the presence of the product of its enzymatic activity, SAM. Cancer cells were treated with candidate MAT2A inhibitors for an appropriate incubation period. The cells were then lysed using a reagent that quenches further enzyme activity. Soluble metabolites containing SAM were collected, and SAM itself was measured directly from the lysate using quantitative LC-MS / MS. Ta.

[0415] The standard assay was performed using the HCT116 human colon cancer cell line (commercially available from Horizon Discovery) that had been genetically engineered to lack the MTAP gene. This was done because the absence of the MTAP gene was found to predict sensitivity to MAT2A inhibitors. The cell line was used. Cells were seeded in 96-well dishes at an appropriate cell density. After 24 hours, the cells were then treated with candidate MAT2A inhibitors. Compounds were first diluted in 100% DMSO before being added to the cells. Serial dilutions are typically made in 3-fold serial dilutions, with 10 dose points, including a DMSO-only control. Then, 5 μL of compound in DMSO was added to 495 μL of cell culture medium. Compounds were transferred to working stock plates in cell culture medium by adding 25 μL of working stock to the cells in 100 μL of culture medium. This working stock was then diluted 5-fold and added to the cells. After addition, the cells were incubated at 37°C / 5% CO2 for 72 hours.

[0416] To quantify SAM levels after compound treatment, cells were gently washed once in ammonium carbonate buffer (75 mM at pH 7.4), placed on dry ice, and extracted with metabolite extraction buffer (80% chilled methanol containing acetic acid at a final concentration of 1 M, with 200 ng / mL deuterated d3-SAM as an internal control). After centrifugation at 3,200 rpm for 30 minutes at 4°C, the supernatant was The samples were collected and stored at −80° C. until analyzed by liquid chromatography with tandem mass spectrometry (LC-MS / MS). The LC-MS / MS analysis was performed in positive ion spray mode. and an API 6500 mass spectrometer (Sciex, Fremont, MA, USA) equipped with a Waters UPLC Acquity (Waters, Milford, MA, USA) BEH Amide column. Multiple reaction monitoring was performed for SAM and d3-SAM standards. The mass transitions at m / z 399.2 → 250.1 and 402.2 → 250.1 were obtained, respectively. A typical LC-MS / MS analysis used an initial flow rate of 25%. Phase A (5:95 (v / v) acetonitrile and HCl containing 1% formic acid and 10 mM ammonium acetate) and water) and 75% mobile phase B (95:5 (v / v) acetonitrile and water containing 1% formic acid and 10 mM ammonium acetate) at 0.5 ml / min, followed by 75% to 35% mobile phase B and 25% to 65% mobile phase B in 0.2–0.5 min. phase A, 0.5 min at 65% mobile phase A and 35% mobile phase B, 1.0–1.1 min at 35%–75% mobile phase B, 65%–25% mobile phase A, and 1.1 min at 25% mobile phase A and 75% mobile phase B for a total run time of 1.5 min.

[0417] Certain compounds disclosed herein were tested in the aforementioned assays and demonstrated that the compounds inhibited the binding of SAMs. Inhibition is scored according to the following IC 50 The concentrations determined were: (A) less than 100 nM (>60% maximal inhibition), (B) 100 nM to 1 μM (>60% maximal inhibition), (C) greater than 1 μM (>60% maximal inhibition), and (NT) not tested, as shown in Table 2 below.

[0418] Cell proliferation inhibition assay

[0419] The effect of test compounds on cancer cell proliferation was assessed by treating cancer cells with the compounds for 4 days, followed by and assessed by measuring proliferation using an ATP-based cell proliferation value (Cell Titer Glo, Promega).

[0420] In a typical assay, isogenic pairs of HCT116 human colon cancer cell lines differing only in MTAP deletion status (HCT116 MTAP+ / + and HCT116 MTAP- / -) were plated at appropriate cell densities in 96-well dishes. After 24 hours, the cells were then treated with a candidate MAT2A inhibitor. Before addition, compounds were first serially diluted in 100% DMSO, typically in 3-fold serial dilutions, starting at a 500-fold higher dose with 10 dose points, including a DMSO-only control. Compounds were then transferred to working stock plates in cell culture medium by adding 5 μL of compound in DMSO to 495 μL of cell culture medium. 25 μL of working stock was then added to 100 μL of culture medium. This working strain was further diluted 5-fold by adding it to cells in culture medium. After compound addition, cells were incubated at 37°C / 5% CO2 for 4 days. .

[0421] To measure inhibition of cell proliferation, cells were allowed to equilibrate to room temperature for 30 min and then 125 μL of Cell The plate was then covered with aluminum foil and shaken for 15 minutes to ensure complete mixing and cell lysis. The luminescence signal was then measured using an ATP standard curve. Measurements were performed using a plate-based luminometer, Veritas version 1.9.2, to confirm assay reproducibility between runs. The luminescence measurements were converted to a proliferation index by subtracting the ATP luminescence signal measured from the bank (cell-free) wells from each data point and dividing by the ATP luminescence signal measured in the 0.2% DMSO control wells, adjusted for the signal in the blank wells. Compound activity was then expressed as the percentage change in proliferation relative to the DMSO control in the plate, versus the log10 of compound concentration in molar (M).

[0422] Certain compounds disclosed herein were tested in the aforementioned assays and were found to inhibit cell proliferation as determined by an IC score according to the following criteria: 50 was determined as shown in Table 2 below: (A) <100 nM (>30% maximal inhibition for MTAP- / -; >10% maximal inhibition for MTAP+ / +), (B) between 100 nM and 1 μM (>30% maximal inhibition for MTAP- / -; >10% maximal inhibition for MTAP+ / +). >10% maximal inhibition), (C) ≥1 μM, and (NT) not tested.

[0423] Table 2 [Table 2-5] [Table 2-6] [Table 2-7] [Table 2-8] [Table 2-9]

Claims

[Claim 1] Azaheterobicyclic inhibitors of Mat2A, and methods of use for the treatment of cancer, etc.

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