Heterocyclic spiro compounds and methods for using them

Abstract

The present disclosure provides compounds of Formula I that have activity as inhibitors of G12C mutant KRAS protein. TIFF2023549055000112.tif49170 The disclosure also provides pharmaceutical compositions comprising the compounds, uses, and methods for treating certain disorders such as cancer, including but not limited to lung cancer, pancreatic cancer, and colorectal cancer.

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims the benefits of the International Patent Application PCT / CN2020 / 123913 filed on 27 October 2020, which is incorporated herein by reference in its entirety.

[0002] This disclosure provides a compound having activity as an inhibitor of the G12C mutant KRAS protein. This disclosure also provides pharmaceutical compositions comprising this compound, uses, and methods for treating certain disorders, including but not limited to cancers, including lung cancer, pancreatic cancer, and colorectal cancer. [Background technology]

[0003] Since its identification in 1982 as one of the first human oncogenes (Der et al., 1982), KRAS (Carsten rat sarcoma virus oncogene homolog) has been the subject of extensive academic and industrial research as a key node in the MAPK signaling pathway, as a transformer in the network of parallel effector pathways (e.g., PI3K / AKT) (Vojtek et al., 1998), and as a potential target for anticancer drugs (Malumbres et al., 2003). Despite advances in the development of inhibitors of upstream and downstream nodes in the MAPK pathway (e.g., EGFR (Sridhar et al., 2003), BRAF (Holderfield et al., 2014), and MEK (Caunt et al., 2015)), the KRAS protein has historically demonstrated resistance to direct inhibition.

[0004] KRAS is a G protein that links extracellular pro-mitotic signaling with intracellular pro-proliferative responses. KRAS acts as an intracellular "on / off" switch. Stimulation by mitogenic factors induces binding of GTP to KRAS, leading to a conformational change that enables KRAS to interact with downstream effector proteins, resulting in cell proliferation. Normally, pro-proliferative signaling is regulated by the action of GTPase-activated protein (GAP), which reverts KRAS to its GDP-bound, non-proliferative state. Mutations in KRAS disrupt this regulated cycling of KRAS between the GDP-bound and GDP-activated states, leading to the accumulation of the GDP-bound active state and dysfunction of cell proliferation (Simanshu et al., 2017).

[0005] Attempts to develop inhibitors of mutant KRAS protein have historically been hampered by the lack of a drug-advantageous pocket on the protein surface (Cox et al., 2014). In 2013, Shokat and his collaborators developed GDP-KRAS G12C KRAS, a commonly known (O'Bryan, 2019) oncogenic variant of KRAS, binds to the previously unrecognized allosteric pocket and inhibits subsequent activation. G12 Covalent inhibitors of KRAS were identified (Ostrem et al., 2013). This discovery represents a significant new development in KRAS inhibitor research, and recently, KRAS inhibitors have entered human clinical trials. See, for example, https: / / clinicaltrials.gov / : for example, NCT03600883&NCT04185883 (AMG510) and NCT03785249 (MRTX849) (last accessed: August 29, 2020). [Prior art documents] [Non-patent literature]

[0006] [Non-Patent Document 1] Der et al., 1982 [Non-Patent Document 2] Vojtek et al., 1998 [Non-Patent Document 3] Malumbres et al., 2003 [Non-Patent Document 4] Sridhar et al., 2003 [Non-Patent Document 5] Holderfield et al., 2014 [Non-Patent Document 6] Caunt et al., 2015 [Non-Patent Document 7] Simanshu et al., 2017 [Non-Patent Document 8] Cox et al., 2014 [Non-Patent Document 9] O'Bryan, 2019 [Non-Patent Document 10] Ostrem et al., 2013 [Overview of the project] [Problems that the invention aims to solve]

[0007] Although some progress has been made, further KRAS is needed to treat conditions such as cancer. G12C The need for inhibitors still exists. [Means for solving the problem]

[0008] Firstly, in this specification, compounds of formula I [ka] Or a pharmaceutically acceptable salt thereof is provided, in the formula, R 1 Each instance of appearance is independent of H and C. 1~4 Alkoxy, -(CH2)-C 1~4is dialkylamino, aziridin-1-yl-methyl, azetidin-1-yl-methyl, pyrrolidin-1-yl-methyl, piperidin-1-yl-methyl, or morpholin-1-yl-methyl, R 2 is H, halogen, -CN, C 1~4 alkyl, C 1~4 haloalkyl, -CH2CN, -CH2OH, C 1-4 alkoxy, or C 1~4 haloalkoxy, optionally, one R 1 and R 2 together with the carbon atom to which they are attached,

Chemical formula

[0009] Secondly, this specification provides pharmaceutical compositions comprising a compound of formula I, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0010] Thirdly, this specification provides for use in the treatment of cancer a compound of formula I as described herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition.

[0011] Here, reference is made in detail to embodiments of the present disclosure. While specific embodiments of the present disclosure are described, it will be understood that the embodiments of the present disclosure are not intended to be limited to these described embodiments. On the contrary, reference to embodiments of the present disclosure is intended to embrace alternative forms, modifications, and equivalents that may be included within the spirit and scope of the embodiments of the present disclosure as defined by the appended claims. **DETAILED DESCRIPTION OF THE INVENTION**

[0012] As Embodiment 1, in the present specification, a compound of Formula I **[Chemical Formula]** or a pharmaceutically acceptable salt thereof is provided, wherein R 1 is, independently for each occurrence, H, C 1~4 alkoxy, -(CH2)-C 1~4 dialkylamino, aziridin-1-yl-methyl, azetidin-1-yl-methyl, pyrrolidin-1-yl-methyl, piperidin-1-yl-methyl, or morpholin-1-yl-methyl, R 2 is H, halogen, -CN, C 1~4 alkyl, C 1~4 haloalkyl, -CH2CN, -CH2OH, C 1-4 alkoxy, or C 1~4 haloalkoxy, Optionally, one R 1 and R 2 together with the carbon atom to which they are attached, **[Chemical Formula]** form a group, R 3 is, independently for each occurrence, H, halogen, CN, OH, -CH2OH, C 1~4 alkyl, C 1~4 haloalkyl, -CH2CN, or C 1~4is an alkoxy, and two substituents R bonded to the same carbon atom 3 optionally, together with the above carbon atom, forms a C 3~6 cycloalkyl or carbonyl group, A, independently for each occurrence, is CR 3 R 3 or absent, R 4 is Z 1 -CH(Z 2 -R 5 )-CH2-R 6 where Z 1 is O, NH, N(C 1~4 alkyl), or CH2, Z 2 is absent or CH2, R 5 is C 1~4 alkyl, C 1~4 haloalkyl, C 3~6 cycloalkyl, C 3~6 heterocycloalkyl, phenyl, or 5- to 6-membered heteroaryl, phenyl is optionally substituted with 1 to 3 substituents selected from halogen, -CN, C 1~3 alkyl, C 1~3 haloalkyl, C 1~3 alkoxy, and C 1~3 haloalkoxy, heteroaryl is optionally substituted with 1 to 3 substituents selected from -CN, C 1~4 alkyl, C 1~4 haloalkyl, C 1~4 alkoxy, and C 1~4 haloalkoxy, R 6 is -CO(NR 7 R 7 ), phenyl, 5,5-dimethyl-3,5-dihydro-4H-imidazol-4-one-2-yl, or 5- to 6-membered heteroaryl, and heteroaryl is optionally -CN, C 1~4 alkyl, C 1~4 haloalkyl, C 1~4 alkoxy, and C 1~4Substituted with 1 to 3 substituents selected from haloalkoxys, R 7 Each instance is independently H or C 1~4 It is alkyl, X 1 CR 8 or N, X 2 is CH or N, X 3 is C or N, X 4 is C or N, R 8 H, halogen, CN, C 1~4 Alkyl, C 1~4 Haloalkyl, C 1~4 Alkoxy, C 1~4 Haloalkoxy, C 3~5 Cycloalkyl, or C 3~5 It is a cyclohaloalkyl, B, together with the atom to which it is bonded, forms a 4-7 membered fully saturated, fully unsaturated, or partially unsaturated carbon ring or heterocyclic system. A heterocyclic system contains 1 to 5 heteroatoms selected from N, O, and S. The ring system has 1 to 5 substituents R, which can be selected arbitrarily. 9 Replaced by, R 9 Each instance is independently represented as halogen, OH, -CN, -NH2, and C(=O)C. 1~6 Alkyl, C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~4 Alkoxy, C 1~4 Haloalkoxy, C 3~5 Cycloalkyl, C 3~5 It is a cyclohaloalkyl, phenyl, or 5-6 member heteroaryl, C 1~6 Alkyls can be optionally -CO(C 1~4 Alkylamino) or -CO(C 1~4 Substituted with dialkylamino, Phenyl is optionally substituted with 1 to 3 independently selected halogens. Heteroaryls can be optionally composed of halogens, C 1~4 Alkyl and C 1~4 Substituted with 1 to 3 substituents selected from haloalkyl groups, Two substituents R 9 Both are optionally -(CH2) n - Forms a group with two substituents R 9 -(CH2) forms a ring with a bonded ring atom or multiple ring atoms. n - The group has one -CH2- group which is optionally substituted with one heteroatom selected from N, O, and S. n is 1, 2, 3, or 4.

[0013] As Embodiment 2, the compound of Embodiment 1, or a pharmaceutically acceptable salt thereof, is provided herein, R 3 Is it not -CN, or, Z 2 R is absent, 5 Is it 2-cyanophenyl, or R 5 Is it not pyrazole-3-yl,2-methyl, or B is [ka] isn't it.

[0014] As Embodiment 3, the compound of Embodiment 1, or a pharmaceutically acceptable salt thereof, is provided herein, where the compound is (3R)-3-(3-cyanophenyl)-N-methyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)propanamide; (3S)-3-((2-((7S)-7-(hydroxymethyl)-2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; 1-(6-(4-(((2S)-4-methyl-1-(1H-pyrazole-3-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; (3S)-3-((2-(8-cyano-2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-3-(2-cyanophenyl)-N-methyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)propanamide; (3R)-3-(2-cyanophenyl)-N-methyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)propanamide; (3S)-N,5-dimethyl-3-((8-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7,8-dihydro-6H-pyrimido[5,4-b][1,4]oxazine-4-yl)amino)hexaneamide; (3S)-N,5-dimethyl-3-((7-(2-propanyl)-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7H-pyrrolo[2,3-d]pyrimidine-4-yl)amino)hexanamide; or (3S)-N,5-dimethyl-3-((7-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7H-purine-6-yl)amino)hexaneamide is not.

[0015] As Embodiment 4, the Specified herein provides the compound of Embodiment 1, or a pharmaceutically acceptable salt thereof, wherein the compound has an IC50 of less than 10 μM in a 2h or 20h bond exchange assay.

[0016] As Embodiment 5, the Specified herein provides one of the compounds from Embodiments 1 to 4, or a pharmaceutically acceptable salt thereof, where Each R 1 H is H.

[0017] As Embodiment 6, the Specified herein provides one of the compounds from Embodiments 1 to 4, or a pharmaceutically acceptable salt thereof, where One R 1 and R 2 These, along with the carbon atoms to which they are bonded, [ka] Forms a base.

[0018] As Embodiment 7, the Specified herein provides one of the compounds from Embodiments 1 to 5, or a pharmaceutically acceptable salt thereof, where R 2 is H, or C 1~4 It is a haloalkyl group.

[0019] As Embodiment 8, the Specified herein provides one of the compounds from Embodiments 1 to 5, or a pharmaceutically acceptable salt thereof, where R 2 This is H or CF3.

[0020] As Embodiment 9, the Specified herein provides one of the compounds from Embodiments 1 to 5, or a pharmaceutically acceptable salt thereof, where R 2 H is H.

[0021] As Embodiment 10, the Specified herein provides one compound from Embodiments 1 to 9, or a pharmaceutically acceptable salt thereof, where R 3 This is H, or a halogen.

[0022] As Embodiment 11, the Specified herein provides one of the compounds from Embodiments 1 to 9, or a pharmaceutically acceptable salt thereof, where R3 It is either H or F.

[0023] As Embodiment 12, the Specified herein provides one of the compounds from Embodiments 1 to 9, or a pharmaceutically acceptable salt thereof, where R 3 H is H.

[0024] As Embodiment 13, the Specified herein provides one compound from Embodiments 1 to 12, or a pharmaceutically acceptable salt thereof, where One A is absent, and the other A is CR 3 R 3 That is the case.

[0025] As Embodiment 14, the Specified herein provides one of the compounds from Embodiments 1 to 12, or a pharmaceutically acceptable salt thereof, where Both A's are absent.

[0026] As Embodiment 15, the Specified herein provides one of the compounds from Embodiments 1 to 12, or a pharmaceutically acceptable salt thereof, where [ka] teeth, [ka] That is the case.

[0027] As Embodiment 16, the Specified herein provides one compound from Embodiments 1 to 12, or a pharmaceutically acceptable salt thereof, where [ka] teeth, [ka] That is the case.

[0028] As Embodiment 17, the Specified herein provides one of the compounds from Embodiments 1 to 12, or a pharmaceutically acceptable salt thereof, where [ka] teeth, [ka] That is the case.

[0029] As Embodiment 18, the Specified herein provides one compound from Embodiments 1 to 17, or a pharmaceutically acceptable salt thereof, where Z 1 It is NH.

[0030] As Embodiment 19, the Specified herein provides one compound from Embodiments 1 to 18, or a pharmaceutically acceptable salt thereof, where Z 2 It is CH2.

[0031] As Embodiment 20, the Specified herein provides one compound from Embodiments 1 to 18, or a pharmaceutically acceptable salt thereof, where Z 2 He is absent.

[0032] As Embodiment 21, the Specified herein provides one of the compounds from Embodiments 1 to 20, or a pharmaceutically acceptable salt thereof, where R 5 C 1~4 It is either alkyl or phenyl, and the phenyl is optionally substituted with -CN.

[0033] As Embodiment 22, the Specified herein provides one of the compounds from Embodiments 1 to 20, or a pharmaceutically acceptable salt thereof, where R 5 It is -CH(CH3)2, phenyl, or 3-cyanophenyl.

[0034] As Embodiment 23, the Specified herein provides one of the compounds from Embodiments 1 to 20, or a pharmaceutically acceptable salt thereof, where R 5 It is -CH(CH3)2.

[0035] As Embodiment 24, the Specified herein provides one of the compounds from Embodiments 1 to 23, or a pharmaceutically acceptable salt thereof, where R 6 is -CO(NR 7 R 7 ), 5,5-dimethyl-3,5-dihydro-4H-imidazole-4-on-2-yl, or a 5-membered heteroaryl, wherein the heteroaryl optionally has 1 to 3 C 1~4 Substituted with alkyl substituents, R 7 Each instance is independently H or C 1~4 It is alkyl.

[0036] As Embodiment 25, the Specified herein provides one of the compounds from Embodiments 1 to 23, or a pharmaceutically acceptable salt thereof, where R 6 -CO(NHR 7 ), or a 5-membered heteroaryl, wherein the heteroaryl optionally contains one C 1~4 Substituted with alkyl substituents, R 7 C 1~4 It is alkyl.

[0037] As Embodiment 26, the Specified herein provides one of the compounds from Embodiments 1 to 23, or a pharmaceutically acceptable salt thereof, where R 6is -CO(NHCH3), 5,5-dimethyl-3,5-dihydro-4H-imidazole-4-on-2-yl, or a 5-membered heteroaryl, where the heteroaryl is pyrazole, imidazole, 1,2,3-triazole, 1,2,4-triazole, 1,2-oxazole, 1,3-oxazole, 1,3,4-oxadiazole, 1,2,4-oxadiazole, 1,3-thiazole, or 1,3,4-thiadiazole, where the heteroaryl optionally has one C 1~4 Substituting with alkyl substituents.

[0038] As Embodiment 27, the Specified herein provides one of the compounds from Embodiments 1 to 23, or a pharmaceutically acceptable salt thereof, where R 6 is -CO(NHCH3) or a five-membered heteroaryl, where the heteroaryl is imidazole, 1,2-oxazole, 1,3,4-oxadiazole, 1,3,4-thiadiazole, or 1,2,3-triazole, and the heteroaryl is optionally substituted with one methyl group.

[0039] As Embodiment 28, the Specified herein provides one of the compounds from Embodiments 1 to 17, or a pharmaceutically acceptable salt thereof, where R 4 teeth, [ka] That is the case.

[0040] As Embodiment 29, the Specified herein provides one compound from Embodiments 1 to 17, or a pharmaceutically acceptable salt thereof, where R 4 teeth, [ka] That is the case.

[0041] As Embodiment 30, the Specified herein provides one compound from Embodiments 1 to 29, or a pharmaceutically acceptable salt thereof, where X 1 CR 8 That is the case.

[0042] As Embodiment 31, the Specified herein provides one compound from Embodiments 1 to 29, or a pharmaceutically acceptable salt thereof, where X 1 It is N.

[0043] As Embodiment 32, the Specified herein provides one compound from Embodiments 1 to 31, or a pharmaceutically acceptable salt thereof, where X 2 It is CH.

[0044] As Embodiment 33, the Specified herein provides one of the compounds from Embodiments 1 to 31, or a pharmaceutically acceptable salt thereof, where X 2 It is N.

[0045] As Embodiment 34, the Specified herein provides one of the compounds from Embodiments 1 to 33, or a pharmaceutically acceptable salt thereof, where X 3 It is C.

[0046] As Embodiment 35, the Specified herein provides one of the compounds from Embodiments 1 to 33, or a pharmaceutically acceptable salt thereof, where X 3 It is N.

[0047] As Embodiment 36, the Specified herein provides one compound from Embodiments 1 to 35, or a pharmaceutically acceptable salt thereof, where X 4 It is C.

[0048] As Embodiment 37, the Specified herein provides one of the compounds from Embodiments 1 to 35, or a pharmaceutically acceptable salt thereof, where X 4 It is N.

[0049] As Embodiment 38, the Specified herein provides one of the compounds from Embodiments 1 to 29, or a pharmaceutically acceptable salt thereof, where X 1 is N, and X 2 is N, and X 3 C is X 4 Is it C, or X 1 is N, and X 2 CH is X 3 C is X 4 Is it C, or X 1 is N, and X 2 is N, and X 3 is N, and X 4 Is it C, or X 1 is N, and X 2 CH is X 3 C is X 4 It is N.

[0050] As Embodiment 39, the Specified herein provides one of the compounds from Embodiments 1 to 29, or a pharmaceutically acceptable salt thereof, where X 1 is N, and X 2 is N, and X 3 C is X 4 Is it C, or X 1 is N, and X 2 CH is X 3 C is X 4 C is C.

[0051] As Embodiment 40, the Specified herein provides one compound from Embodiments 1 to 29, or a pharmaceutically acceptable salt thereof, where X 1 is N, and X 2 is N, and X 3 C is X 4 C is C.

[0052] As Embodiment 41, this specification provides one compound from Embodiments 1-34, 36, and 38-40, or a pharmaceutically acceptable salt thereof, where B, along with the atom it is bonded to, [ka] Form a ring system selected from, The ring system has 1 to 5 substituents R, which can be selected arbitrarily. 9 It will be replaced with.

[0053] As Embodiment 42, this specification provides one compound from Embodiments 1-34, 36, and 38-40, or a pharmaceutically acceptable salt thereof, where B, along with the atom it is bonded to, [ka] Form a ring system selected from, The ring system has 1 to 5 substituents R, which can be selected arbitrarily. 9 It will be replaced with.

[0054] As Embodiment 43, this specification provides one compound from Embodiments 1-34, 36, and 38-40, or a pharmaceutically acceptable salt thereof, where B, along with the atom it is bonded to, [ka] It forms a ring system selected from among them.

[0055] As Embodiment 44, the Specified herein provides one of the compounds from Embodiments 1 to 42, or a pharmaceutically acceptable salt thereof. The ring system has 1 to 2 substituents R, which can be selected optionally. 9 Replaced by, R 9 Each instance is independently determined as follows: halogen, -CN, C(=O)C 1~6 Alkyl, C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~4Alkoxy, C 3~5 They are cycloalkyl or 5-6 membered heteroaryl compounds.

[0056] As Embodiment 45, the Specified herein provides one of the compounds from Embodiments 1 to 42, or a pharmaceutically acceptable salt thereof. The ring system has 1 to 2 substituents R, which can be selected optionally. 9 Replaced by, R 9 Each instance is independent of C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~4 Alkoxy, C 3~5 It is a cycloalkyl or a 5-membered heteroaryl.

[0057] As Embodiment 46, the Specified herein provides one compound from any of Embodiments 1 to 42, or a pharmaceutically acceptable salt thereof. The ring system has 1 to 2 substituents R, which can be selected optionally. 9 Replaced by, R 9 Each of these is independently Cl, -CN, acetyl, methyl, isopropyl, trifluoromethyl, methoxy, cyclopropyl, or 1,3-thiazolyl.

[0058] As Embodiment 47, the Specified herein provides one of the compounds from Embodiments 1 to 42, or a pharmaceutically acceptable salt thereof, where The ring system has 1 to 2 substituents R, which can be selected optionally. 9 Replaced by, R 9 Each of these is independently methyl, isopropyl, trifluoromethyl, methoxy, cyclopropyl, or 1,3-thiazolyl.

[0059] As Embodiment 48, the compound of Embodiment 1, or a pharmaceutically acceptable salt thereof, is provided herein, where the compound is (S)-3-((2-(2-acryloyl-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydroquinazoline-4-yl)amino)-N,5-dimethylhexaneamide; (3S)-3-((2-(8,8-difluoro-2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7-methyl-5,6,7,8-tetrahydro-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-N,5-dimethyl-3-((7-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)hexaneamide; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-6,7-dihydro-5H-cyclopenta[d]pyrimidine-4-yl)amino)hexaneamide; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)hexaneamide; (3S)-N,5-dimethyl-3-((3(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-1-isoquinolinyl)amino)hexaneamide; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)flo[3,2-d]pyrimidine-4-yl)amino)hexaneamide; (3S)-N,5-dimethyl-3-(((8R)-8-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)hexanamide; (3S)-3-((6-acetyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3,4]octan-6-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidine-4-yl)amino)-N,5-dimethylhexaneamide; (3S)-N,5-dimethyl-3-((2-methyl-5-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-2H-pyrazolo[4,3-d]pyrimidine-7-yl)amino)hexaneamide; (3S)-N,5-dimethyl-3-((5-(2-(2-propenoyl)-2,6-diazaspiro[3,4]octan-6-yl)[1,3]thiazolo[5,4-d]pyrimidine-7-yl)amino)hexanamide; 5,5-dimethyl-2-((2S)-4-methyl-2-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)pentyl)-3,5-dihydro-4H-imidazole-4-one; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,7-dihydrofl[3,4-d]pyrimidine-4-yl)amino)hexaneamide; (3S)-N,5-dimethyl-3-(((8S)-8-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)hexaneamide; (3S)-N,5-dimethyl-3-((7-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7H-pyrrolo[2,3-d]pyrimidine-4-yl)amino)hexaneamide; (3S)-N,5-dimethyl-3-((9-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-9H-purine-6-yl)amino)hexanamide; (3S)-3-((2-(8,8-difluoro-2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; 1-(6-(4-(((2S)-4-methyl-1-(4H-1,2,4-triazol-3-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; (3S)-N,5-dimethyl-3-((7-(2-propanyl)-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)hexaneamide; (3S)-3-(3-cyanophenyl)-N-methyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)propanamide; 1-(6-(4-(((2S)-4-methyl-1-(5-methyl-1,3,4-oxadiazole-2-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(4-(((2S)-1-(1H-imidazole-2-yl)-4-methyl-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; (3S)-3-((2-((7R)-7-(hydroxymethyl)-2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-N,5-dimethyl-3-((7-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)hexaneamide; (3S)-3-((7-cyano-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-3-((2-(8-fluoro-2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-3-((2-(8-fluoro-2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7-methyl-5,6,7,8-tetrahydro-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-3-((7,7-dimethyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-3-((7-chloro-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3,4]octan-6-yl)-7,8-dihydro-6H-pyrano[3,2-d]pyrimidine-4-yl)amino)hexanamide; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7-(trifluoromethyl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)hexaneamide; (3S)-N,5-dimethyl-3-((6-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)hexaneamide; (3S)-N,5-dimethyl-3-((7-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)pyrido[3,2-d]pyrimidine-4-yl)amino)hexaneamide; (3S)-N,5-Dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7-(trifluoromethyl)-4-quinazolinyl)amino)hexanamide; (3S)-N,5-Dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7-(1,3-thiazol-2-yl)pyrido[3,2-d]pyrimidin-4-yl)amino)hexanamide; (3S)-N,5-Dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)pyrido[3,2-d]pyrimidin-4-yl)amino)hexanamide; (3S)-3-((7-Cyclopropyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)pyrido[3,2-d]pyrimidin-4-yl)amino)-N,5-dimethylhexanamide; 1-(6-(7,7-Dimethyl-4-(((2S)-4-methyl-1-(3-methyl-1,2,4-oxadiazol-5-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; (3S)-3-((7-Cyclopropyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)-N,5-dimethylhexanamide; (3S)-N,5-Dimethyl-3-((6-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)hexanamide; (3S)-3-((7-Methoxy-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)pyrido[3,2-d]pyrimidin-4-yl)amino)-N,5-dimethylhexanamide; 1-(6-(7,7-Dimethyl-4-((((2S)-4-methyl-1-(1,3-thiazol-2-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-Dimethyl-4-((((2S)-4-methyl-1-(5-methyl-1,2,4-oxadiazol-3-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-Dimethyl-4-((((2S)-4-methyl-1-(1H-1,2,4-triazol-1-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-Dimethyl-4-((((2S)-4-methyl-1-(1H-pyrazol-1-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-Dimethyl-4-((((2S)-4-methyl-1-(1H-1,2,3-triazol-1-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-Dimethyl-4-((((2S)-4-methyl-1-(1,2-oxazol-3-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-Dimethyl-4-((((2S)-4-methyl-1-(3-methyl-1,2-oxazol-5-yl)-z-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; (3S)-3-((7,7-dimethyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)-N-methyl-4-phenylbutanamide; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(5-methyl-1,3,4-thiadiazole-2-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(5-methyl-1,3,4-oxadiazole-2-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(5-methyl-1,2-oxazol-3-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(4-(((2S)-1-(1H-imidazole-2-yl)-4-methyl-2-pentanyl)amino)-7,7-dimethyl-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(1H-1,2,3-triazol-4-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)pyrido[3,4-d]pyrimidine-4-yl)amino)hexaneamide; 1-(6-(4-(((2S)-4-methyl-1-(1,3-oxazol-2-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; (3S)-N,5-dimethyl-3-((7-methyl-2-(2-(2-(trifluoromethyl)-2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)hexaneamide; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7-(1,3-thiazole-2-yl)-4-quinazolinyl)amino)hexanamide; or It is (S)-2-((2-(2-acryloyl-2,6-diazaspiro[3,4]octan-6-yl)-5,6,7,8-tetrahydroquinazoline-4-yl)amino)-N,4-dimethylpentanamide.

[0060] As Embodiment 49, the compound of Embodiment 1, or a pharmaceutically acceptable salt thereof, is provided herein, where this compound is (3S)-3-((2-(8,8-difluoro-2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7-methyl-5,6,7,8-tetrahydro-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-N,5-dimethyl-3-((7-(2-propanyl)-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)hexaneamide; (3S)-3-((7,7-dimethyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7-(trifluoromethyl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)hexaneamide; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7-(1,3-thiazole-2-yl)pyrido[3,2-d]pyrimidine-4-yl)amino)hexaneamide; (3S)-3-((7-cyclopropyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)pyrido[3,2-d]pyrimidine-4-yl)amino)-N,5-dimethylhexaneamide; (3S)-3-((7-Cyclopropyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-3-((7-Methoxy-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)pyrido[3,2-d]pyrimidine-4-yl)amino)-N,5-dimethylhexaneamide; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(1,2-oxazol-3-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(3-methyl-1,2-oxazol-5-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(5-methyl-1,3,4-thiadiazole-2-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(5-methyl-1,3,4-oxadiazole-2-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(5-methyl-1,2-oxazol-3-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(4-(((2S)-1-(1H-imidazole-2-yl)-4-methyl-2-pentanyl)amino)-7,7-dimethyl-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(1H-1,2,3-triazol-4-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; or It is (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7-(1,3-thiazole-2-yl)-4-quinazolinyl)amino)hexaneamide.

[0061] The foregoing statements merely summarize certain aspects of the disclosure and are not intended to limit, nor should they be interpreted as limiting, the disclosure.

[0062] Composition and route of administration In the disclosed uses, the compounds disclosed herein may be administered alone, but the administered compounds typically exist as active ingredients in a pharmaceutical composition. Accordingly, in one embodiment, the herein provides a pharmaceutical composition comprising the compounds disclosed herein in combination with one or more pharmaceutically acceptable excipients and, optionally, other active ingredients. For example, see Remington: The Science and Practice of Pharmacy, Volume I and Volume II, 20-second edition, edited by Loyd V. Allen Jr., Philadelphia, PA, Pharmaceutical Press, 2012; Pharmaceutical Dosage Forms (Vol. 1-3), Liberman et al., Eds., Marcel Dekker, New York, NY, 1992; Handbook of Pharmaceutical Excipients (3rd Ed.), edited by Arthur H. Kibbe, American Pharmaceutical Association, Washington, 2000; Pharmaceutical Formulation: The Science and Technology of Dosage Forms (Drug Discovery), first edition, edited by GD Tovey, Royal Society of Chemistry, 2018. In one embodiment, the pharmaceutical composition comprises a therapeutically effective amount of the compound disclosed herein.

[0063] The compounds disclosed herein may be administered by any preferred route of administration in the form of a pharmaceutical composition adapted to such route and in doses effective for the treatment of the intended purpose. The compounds and compositions provided herein may be administered, for example, orally, mucosally, topically, transdermally, rectally, pulmonaryly, parenterally, intranasally, intravascularly, intravenously, intraarterially, intraperitoneally, intrathecally, subcutaneously, sublingually, intramuscularly, intrasternally, intravaginally, or by injectable techniques in drug unit formulations containing conventionally pharmaceutically acceptable excipients.

[0064] The pharmaceutical composition can be in the form of, for example, tablets, chewable tablets, mini - tablets, caplets, pills, beads, hard capsules, soft capsules, gelatin capsules, granules, powders, lozenges, patches, creams, gels, sachets, microneedle arrays, syrups, flavored syrups, juices, drops, injection solutions, emulsions, microemulsions, ointments, aerosols, aqueous suspensions, or oily suspensions. The pharmaceutical composition is usually prepared in the form of dosage units containing a specific amount of the active ingredient.

[0065] As Embodiment 50, provided herein is a pharmaceutical composition comprising any one of the compounds of Embodiments 1 - 49, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0066] As Embodiment 51, provided herein is any one of the compounds of Embodiments 1 - 49, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of Embodiment 50 for use as a medicament.

[0067] Method of Use As described herein (see the section entitled "Definitions"), the compounds described herein are understood to include all stereoisomers, tautomers, or pharmaceutically acceptable salts of any of the above. Thus, the methods and uses provided in this disclosure should be understood to also encompass methods and uses employing all such forms.

[0068] In addition to being useful for the treatment of humans, the compounds provided herein can be useful for veterinary treatment of companion animals, exotic animals, and livestock, including mammals, rodents, etc. For example, animals including horses, dogs, and cats can be treated with the compounds provided herein.

[0069] Monotherapy In one embodiment, the Disclosure provides a method of using the compounds or pharmaceutical compositions of the Disclosure to treat disease conditions including, but not limited to, conditions associated with mutations in KRAS G12C (e.g., cancer). See, for example, the sections from paragraph 198, line 1 to paragraph 201, line 36 of U.S. Patent No. 10,519,146B2 issued December 31, 2019, which are incorporated herein by reference.

[0070] While not bound by any particular theory, please note the following: AMG510, like the compounds disclosed herein, is KRAS G12C It is a small molecule that specifically and irreversibly inhibits [AMG510] (Hong et al., 2020, pp. 1208). Hong et al. reported that "preclinical studies showed that [AMG510] inhibited almost all detectable phosphorylation of extracellular signal-regulated kinases (ERKs), key downstream effectors of KRAS, resulting in sustained complete tumor regression in mice with KRAS p.G12C tumors." (See also the section titled "Biological Evaluation" in the same book, and Canon et al., 2019 and Lanman et al., 2020).

[0071] AMG510 was evaluated in a phase 1 dose-escalation and expansion study involving 129 patients with locally advanced or metastatic cancer containing histologically confirmed KRAS G12C mutations, identified by local molecular examination of tumor tissue, including 59 patients with non-small cell lung cancer, 42 patients with colorectal cancer, and 28 patients with other tumor types (Hong et al., 2020, pp. 1208-1209). Hong et al. reported disease control rates (95% CI) of 88.1% in non-small cell lung cancer, 73.8% in colorectal cancer, and 75.0% in other tumor types (Hong et al., 2020, p. 1213, Table 3). In conclusion, the cancer types that showed either stable disease (SD) or partial reduction (PR) as reported by Hong et al. were non-small cell lung cancer, colorectal cancer, pancreatic cancer, appendiceal cancer, endometrial cancer, esophageal cancer, cancer of unknown primary origin, ampulla cancer, gastric cancer, small intestine cancer, paranasal sinus cancer, bile duct cancer, or melanoma (Hong et al., 2020, p. 1212 (Figure A), and Appendix (p. 59 (Figure S5) and p. 63 (Figure S6)).

[0072] KRAS G12C mutations occur in the change cycle shown in the table below (Cerami et al., 2012; Gao et al., 2013). For example, the table shows that 11.6% of subjects with non-small cell lung cancer have cancer in which one or more cells express the KRAS G12C mutant protein. Therefore, KRAS G12C The compounds provided herein (see the section titled “Biological Evaluation” below), which bind specifically and irreversibly to [the specified substance], are useful for the treatment of subjects with cancer, including but not limited to the cancers listed in the table below.

[0073] [Table 1]

[0074] As Embodiment 52, the Specified herein provides one compound from Embodiments 1 to 49, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition from Embodiment 50, for use in the treatment of cancer.

[0075] As Embodiment 53, the Specified herein provides one compound from Embodiments 1 to 49, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition from Embodiment 50, for use in the treatment of cancer in which one or more cells express the KRAS G12C mutant protein.

[0076] As Embodiment 54, the Specified herein provides a compound or pharmaceutical composition for use in Embodiment 52 or 53, wherein the cancer is non-small cell lung cancer, small intestine cancer, appendiceal cancer, colorectal cancer, cancer of unknown primary origin, endometrial cancer, mixed-type cancer of unknown primary origin, pancreatic cancer, hepatobiliary cancer, small cell lung cancer, cervical cancer, germ cell cancer, ovarian cancer, gastrointestinal neuroendocrine cancer, bladder cancer, myelodysplastic / myeloproliferative neoplasm, head and neck cancer, esophageal and gastric cancer, soft tissue sarcoma, mesothelioma, thyroid cancer, leukemia, or melanoma.

[0077] Embodiment 55 provides the use of any one compound from Embodiments 1 to 49, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of Embodiment 50, in the preparation of a drug for treating cancer.

[0078] Embodiment 56, as provided herein, involves the use of any one compound from Embodiments 1 to 49, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of Embodiment 50, in the preparation of a drug for treating cancer in which one or more cells express the KRAS G12C mutant protein.

[0079] Embodiment 57 provides the use of Embodiment 55 or 56, wherein the cancer is non-small cell lung cancer, small intestine cancer, appendiceal cancer, colorectal cancer, cancer of unknown primary origin, endometrial cancer, mixed-type cancer of unknown primary origin, pancreatic cancer, hepatobiliary cancer, small cell lung cancer, cervical cancer, germ cell cancer, ovarian cancer, gastrointestinal neuroendocrine cancer, bladder cancer, myelodysplastic / myeloproliferative neoplasm, head and neck cancer, esophageal and gastric cancer, soft tissue sarcoma, mesothelioma, thyroid cancer, leukemia, or melanoma.

[0080] Embodiment 58, as provided herein, is a method for treating cancer in a subject requiring cancer treatment, comprising administering to the subject a therapeutically effective amount of one compound from Embodiments 1 to 49, or a pharmaceutically acceptable salt thereof.

[0081] Embodiment 59, as provided herein, is a method for treating cancer in a subject in need of cancer treatment, wherein one or more cells express the KRAS G12C mutant protein, the method comprising administering to the subject a therapeutically effective amount of one of the compounds from Embodiments 1 to 49, or a pharmaceutically acceptable salt thereof.

[0082] As Embodiment 60, the Specified herein provides the method of Embodiment 58 or 59, wherein the cancer is non-small cell lung cancer, small intestine cancer, appendiceal cancer, colorectal cancer, cancer of unknown primary origin, endometrial cancer, mixed-type cancer of unknown primary origin, pancreatic cancer, hepatobiliary cancer, small cell lung cancer, cervical cancer, germ cell cancer, ovarian cancer, gastrointestinal neuroendocrine cancer, bladder cancer, myelodysplastic / myeloproliferative neoplasm, head and neck cancer, esophageal and gastric cancer, soft tissue sarcoma, mesothelioma, thyroid cancer, leukemia, or melanoma.

[0083] As Embodiment 61, the method of Embodiment 58 or 59 is provided herein, wherein the cancer is non-small cell lung cancer, colorectal cancer, pancreatic cancer, appendiceal cancer, endometrial cancer, esophageal cancer, cancer of unknown primary origin, ampulla cancer, gastric cancer, small intestine cancer, paranasal sinus cancer, bile duct cancer, or melanoma.

[0084] As Embodiment 62, the method of Embodiment 61 is provided herein, wherein the lung cancer is non-small cell lung cancer.

[0085] As Embodiment 63, the method of Embodiment 61 is provided herein, wherein the lung cancer is colorectal cancer.

[0086] As Embodiment 64, the method of Embodiment 61 is provided herein, wherein lung cancer is pancreatic cancer.

[0087] Embodiment 65 provides a method according to any one of Embodiments 58 to 64, wherein the subject has cancer determined to have one or more cells expressing the KRAS G12C mutant protein prior to administration of the compound or a pharmaceutically acceptable salt thereof.

[0088] Combination therapy The disclosure of the present invention also provides methods of combination therapy in which agents known to modulate other pathways, or other components of the same pathway, or even overlapping sets of target enzymes are used in combination with the compounds of the Disclosure or pharmaceutically acceptable salts thereof. In one embodiment, such therapies include, but are not limited to, combinations of one or more compounds of the Disclosure with chemotherapeutic agents, therapeutic antibodies, and radiotherapy to produce a synergistic or additive therapeutic effect. See, for example, sections 201 (line 37) to 212 (line 46) and 219 (line 64) to 220 (line 39) of U.S. Patent No. 10,519,146B2 issued December 31, 2019, which is incorporated herein by reference.

[0089] Embodiment 66 further comprises the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is an aurora kinase A inhibitor, an AKT inhibitor, an arginase inhibitor, a CDK4 / 6 inhibitor, an ErbB family inhibitor, an ERK inhibitor, a FAK inhibitor, an FGFR inhibitor, a glutaminase inhibitor, an IGF-1R inhibitor, a KIF18A inhibitor, an MCL-1 inhibitor, a MEK inhibitor, an mTOR inhibitor, a PD-1 inhibitor, a PD-L1 inhibitor, a PI3K inhibitor, a Raf kinase inhibitor, a SHP2 inhibitor, a SOS1 inhibitor, a Src kinase inhibitor, or one or more chemotherapeutic agents, as provided herein, one of the methods of Embodiments 58 to 65.

[0090] In one embodiment, the second compound is administered as a pharmaceutically acceptable salt. In another embodiment, the second compound is administered as a pharmaceutical composition comprising the second compound or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0091] Aurora kinase A inhibitors This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is an aurora kinase A inhibitor.

[0092] Examples of Aurora kinase A inhibitors for use in the methods provided herein include alisertib, cenisertib, danusertib, tozasertib, LY3295668(2R,4R)-1-[(3-chloro-2-fluorophenyl)methyl]-4-[[3-fluoro-6-[(5-methyl-1H-pyrazole-3-yl)amino]pyridine-2-yl]methyl]-2-methylpiperidine-4-carboxylic acid), and ENMD-20. 76(6-(4-methylpiperazine-1-yl)-N-(5-methyl-1H-pyrazole-3-yl)-2-[(E)-2-phenylethenyl]pyrimidine-4-amine), TAK-901(5-(3-ethylsulfonylphenyl)-3,8-dimethyl-N-(1-methylpiperidine-4-yl)-9H-pyrido[2,3-b]indole-7-carboxamide), TT-00420(4-[9-(2-chlorophenyl)-6-methyl-2,4,5,8,12-pentazatricyclo[8.4.0.03,7]tetradeca-1(14),3 ,6,8,10,12-hexaen-13-yl]morpholine), AMG900 (N-[4-[3-(2-aminopyrimidine-4-yl)pyridine-2-yl]oxyphenyl]-4-(4-methylthiophen-2-yl)phthalazine-1-amine), MLN8054 (4-[[9-chloro-7-(2,6-difluorophenyl)-5H-pyrimido[5,4-d][2]benzazepine-2-yl]amino]benzoic acid), PF-03814735 (N-[2-[(1R,8S)-4-[[4-(cyclobutylamino)-5-(trifluoro Examples include, but are not limited to, methyl)pyrimidine-2-yl]amino]-11-azatricyclo[6.2.1.02,7]undeca-2(7),3,5-trien-11-yl]-2-oxoethyl]acetamide), SNS-314 (1-(3-chlorophenyl)-3-[5-[2-(thieno[3,2-d]pyrimidine-4-ylamino)ethyl]-1,3-thiazole-2-yl]urea), CYC116 (4-methyl-5-[2-(4-morpholine-4-ylanilino)pyrimidine-4-yl]-1,3-thiazole-2-amine), TAS-119, BI 811283, and TTP607.

[0093] AKT inhibitors This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is an AKT inhibitor.

[0094] Examples of AKT inhibitors for use in the methods provided herein include afuresertib, capivasertib, ipatasertib, uprosertib, BAY1125976 (2-[4-(1-aminocyclobutyl)phenyl]-3-phenylimidazo[1,2-b]pyridazine-6-carboxamide), ARQ092 (3-[3-[4-(1-aminocyclobutyl)phenyl]-5-phenylimidazo[4,5-b]pyridin-2-yl]pyridin-2-amine), and MK2206 (8-[4-(1-aminocyclobutyl)phenyl]-9-phenyl-2H-[1,2,4]tri Examples include, but are not limited to, azolo[3,4-f][1,6]naphthyridine-3-one), SR13668 (indro[2,3-b]carbazole-2,10-dicarboxylic acid, 5,7-dihydro-6-methoxy-,2,10-diethyl ester), ONC201 (11-benzyl-7-[(2-methylphenyl)methyl]-2,5,7,11-tetrazatricyclo[7.4.0.02,6]trideca-1(9),5-dien-8-one), ARQ751 (N-(3-aminopropyl)-N-[(1R)-1-(3-anilino-7-chloro-4-oxoquinazolin-2-yl)buty-3-nyl]-3-chloro-2-fluorobenzamide), RX-0201, and LY2780301.

[0095] Arginase inhibitors This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is an arginase inhibitor.

[0096] Examples of arginase inhibitors for use in the methods provided herein include, but are not limited to, numidargistat and CB280.

[0097] CDK4 / 6 inhibitors This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is a CDK4 / 6 inhibitor.

[0098] As used herein, the terms "CDK4 / 6" refer to cyclin-dependent kinases (CDKs) 4 and 6, which are members of the mammalian serine / threonine protein kinases.

[0099] As used herein, the term “CDK4 / 6 inhibitor” refers to a compound capable of negatively modulating or inhibiting all or part of the enzymatic activity of CDK4 and / or 6.

[0100] Examples of CDK4 / 6 inhibitors for use in the methods provided herein include, but are not limited to, abemaciclib, palbociclib, ribociclib, trilaciclib, and PF-06873600 ((pyrido[2,3-d]pyrimidine-7(8H)-one, 6-(difluoromethyl)-8-[(1R,2R)-2-hydroxy-2-methylcyclopentyl]-2-[[1-(methylsulfonyl1)-4-piperidinyl]amino]).

[0101] In one embodiment, the CDK4 / 6 inhibitor is palbociclib.

[0102] ErbB family inhibitors This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is an ErbB family inhibitor.

[0103] As used herein, the term “ErbB family” refers to the members of the mammalian transmembrane protein tyrosine kinase family, including: ErbB1 (EGFR HER1), ErbB2 (HER2), ErbB3 (HER3), and ErbB4 (HER4).

[0104] As used herein, the term “ErbB family inhibitor” refers to a drug, such as a compound or antibody, that can negatively modulate or inhibit all or part of the activity of at least one member of the ErbB family. Modulation or inhibition of one or more ErbB tyrosine kinases may occur through the modulation or inhibition of the kinase enzyme activity of one or more ErbB family members, or by blocking homodimerization or heterodimerization of ErbB family members.

[0105] In one embodiment, the ErbB family inhibitor is an EGFR inhibitor, such as an anti-EGFR antibody. Exemplary anti-EGFR antibodies for use in the methods provided herein include, but are not limited to, saltumumab, nimotuzumab, matuzumab, nesitumumab, panitumumab, and cetuximab. In one embodiment, the anti-EGFR antibody is cetuximab. In one embodiment, the anti-EGFR antibody is panitumumab.

[0106] In another embodiment, the ErbB family inhibitor is a HER2 inhibitor, such as an anti-HER2 antibody. Exemplary anti-HER2 antibodies for use in the methods provided herein include, but are not limited to, pertuzumab, trastuzumab, and trastuzumab emtansine.

[0107] In yet another embodiment, the ErbB family inhibitor is a HER3 inhibitor, such as an anti-HER3 antibody like HMBD-001 (Hummingbird Bioscience).

[0108] In one embodiment, the ErbB family inhibitor is a combination of an anti-EGFR antibody and an anti-HER2 antibody.

[0109] In one embodiment, the ErbB family inhibitor is an irreversible inhibitor. Examples of irreversible ErbB family inhibitors for use in the manner provided herein include, but are not limited to, afatinib, dacomitinib, canertinib, poziotinib, AV412 ((N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[3-methyl-3-(4-methyl-1-piperazinyl)-1-butin-1-yl]-6-quinazolinyl]-2-propenamide)), PF6274484 ((N-[4-[(3-chloro-4-fluorophenyl)amino]-7-methoxy-6-quinazolinyl]-2-propenamide) and HKI357 ((E)-N-[4-[3-chloro-4-[(3-fluorophenyl)methoxy]anilino]-3-cyano-7-ethoxyquinoline-6-yl]-4-(dimethylamino)buto-2-enamide).

[0110] In one embodiment, the irreversible ErbB family inhibitor is afatinib. In another embodiment, the irreversible ErbB family inhibitor is dacomitinib.

[0111] In one embodiment, the ErbB family inhibitors are reversible inhibitors. Examples of reversible ErbB family inhibitors for use in the methods provided herein include erlotinib, gefitinib, sapitinib, valitinib, tarloxotinib, TAK-285 (N-(2-(4-((3-chloro-4-(3-(trifluoromethyl)phenoxy)phenyl)amino)-5H-pyrrolo[3,2-d]pyrimidine-5-yl)ethyl)-3-hydroxy-3-methylbutanamide), AEE788 ((S)-6-(4-((4-ethylpiperazine-1-yl)methyl)phenyl)-N-(1-phenyl Examples include, but are not limited to, ((3S)-3-morpholinylmethyl-[4-[[1-[(3-fluorophenyl)methyl]-1H-indazole-5-yl]amino]-5-methylpyrrolo[2,1-f][1,2,4]triazine-6-yl]carbamate, and GW583340(N-[3-chloro-4-[(3-fluorophenyl)methoxy]phenyl]-6-[2-[(2-methylsulfonylethylamino)methyl]-1,3-thiazole-4-yl]quinazoline-4-amine).

[0112] In one embodiment, the reversible ErbB family inhibitor is sapitinib. In another embodiment, the reversible ErbB family inhibitor is tarloxotinib.

[0113] ERK inhibitors This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is an ERK inhibitor.

[0114] Examples of ERK inhibitors for use in the methods provided herein include urixertinib, ravoxertinib, CC-90003 (N-[2-[[2-[(2-methoxy-5-methylpyridine-4-yl)amino]-5-(trifluoromethyl)pyrimidine-4-yl]amino]-5-methylphenyl]propaneamide), and LY3214996 (6,6-dimethyl-2-[2- Examples include, but are not limited to, [(2-methylpyrazole-3-yl)amino]pyrimidine-4-yl]-5-(2-morpholine-4-ylethyl)thieno[2,3-c]pyrrole-4-one), KO-947 (1,5,6,8-tetrahydro-6-(phenylmethyl)-3-(4-pyridinyl)-7H-pyrazolo[4,3-g]quinazolin-7-one), ASTX029, LTT462, and JSI-1187.

[0115] FAK inhibitors This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is a FAK inhibitor.

[0116] Examples of FAK inhibitors for use in the methods provided herein include, but are not limited to, GSK2256098 (2-[[5-chloro-2-[(5-methyl-2-propane-2-ylpyrazole-3-yl)amino]pyridine-4-yl]amino]-N-methoxybenzamide), PF-00562271 (N-methyl-N-[3-[[[2-[(2-oxo-1,3-dihydroindole-5-yl)amino]-5-(trifluoromethyl)pyrimidine-4-yl]amino]methyl]pyridine-2-yl]methanesulfonamide), VS-4718 (2-[[2-(2-methoxy-4-morpholine-4-ylanilino)-5-(trifluoromethyl)pyridine-4-yl]amino]-N-methylbenzamide), and APG-2449.

[0117] FGFR inhibitors This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is an FGFR inhibitor.

[0118] Examples of FGFR inhibitors for use in the methods provided herein include futibatinib, pemigatinib, ASP5878 (2-[4-[[5-[(2,6-difluoro-3,5-dimethoxyphenyl)methoxy]pyrimidine-2-yl]amino]pyrazole-1-yl]ethanol), AZD4547 (N-[5-[2-(3,5-dimethoxyphenyl)ethyl]-1H-pyrazole-3-yl]-4-[(3S,5R)-3,5-dimethylpiperazine-1-yl]benzamide), and Debio1347 ([5-amino Examples include, but are not limited to, -1-(2-methyl-3H-benzimidazole-5-yl)pyrazole-4-yl]-(1H-indole-2-yl)methanone), INCB062079, H3B-6527(N-[2-[[6-[(2,6-dichloro-3,5-dimethoxyphenyl)carbamoyl-methylamino]pyrimidine-4-yl]amino]-5-(4-ethylpiperazine-1-yl)phenyl]propa-2-enamide), ICP-105, CPL304110, HMPL-453, and HGS1036.

[0119] Glutaminase inhibitors This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is a glutaminase inhibitor.

[0120] Examples of glutaminase inhibitors for use in the methods provided herein include, but are not limited to, teragrenastat, IPN60090, and OP330.

[0121] IGF-1R inhibitors This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is an IGF-1R inhibitor.

[0122] Examples of IGF-1R inhibitors for use in the methods provided herein include cictumumab, darotuzumab, lincitinib, ganitumab, lobatumumab, and BMS-754807((2S)-1-[4-[(5-cyclopropyl-1H-pyrazole-3-yl)amino]pyrrolo[2,1-f][1,2,4]triazine-2-yl]-N-(6-fluoropyridine-3-yl) Examples include, but are not limited to, )-2-methylpyrrolidine-2-carboxamide), KW-2450(N-[5-[[4-(2-hydroxyacetyl)piperazine-1-yl]methyl]-2-[(E)-2-(1H-indazole-3-yl)ethenyl]phenyl]-3-methylthiophene-2-carboxamide), PL225B, AVE1642, and BIIB022.

[0123] KIF18A inhibitors This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is a KIF18A inhibitor.

[0124] Examples of KIF18A inhibitors for use in the manner provided herein include, but are not limited to, those disclosed in U.S. Patent Application Publication No. 2020 / 0239441, International Publication Brochure No. 2020 / 132649, International Publication Brochure No. 2020 / 132651, and International Publication Brochure No. 2020 / 132653, each of which is incorporated herein by whole reference.

[0125] MCL-1 inhibitors This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is an MCL-1 inhibitor.

[0126] Examples of MEK inhibitors for use in the methods provided herein include murizatoclax, tapotoclax, and AZD5991((3aR)-5-chloro-2,11,12,24,27,29-hexahydro-2,3,24,33-tetramethyl-22H-9,4,8-(metheniminomethyno))-14,20:26,23-dimetheno-10H,20H-pyrazolo[4,3-1][2,15 Examples include, but are not limited to, [22,18,19]benzooxaditiasiazacyclohexacosin-32-carboxylic acid), MIK665 ((αR)-α-[[(5S)-5-[3-chloro-2-methyl-4-[2-(4-methyl-1-piperazinyl)ethoxy]phenyl]-6-(4-fluorophenyl)thieno[2,3-d]pyrimidine-4-yl]oxy]-2-[[2-(2-methoxyphenyl)-4-pyrimidinyl]methoxy]benzenepropanoic acid), and ABBV-467.

[0127] In one embodiment, the MCL-1 inhibitor is murizatocrax. In another embodiment, the MCL-1 inhibitor is tapotocrax.

[0128] MEK inhibitors This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is a MEK inhibitor.

[0129] Examples of MEK inhibitors for use in the methods provided herein include trametinib, cobimetinib, selumetinib, pimacertib, refametinib, PD-325901 (N-[(2R)-2,3-dihydroxypropoxy]-3,4-difluoro-2-(2-fluoro-4-iodoanilino)benzamide), AZD8330 (2-(2-fluoro-4-iodoanilino)-N-(2-hydroxyethoxy)-1,5-dimethyl-6-oxopyridine-3-ka Luboxamide), GDC-0623 (5-(2-fluoro-4-iodoanilino)-N-(2-hydroxyethoxy)imidazo[1,5-a]pyridine-6-carboxamide), RO4987655 (3,4-difluoro-2-(2-fluoro-4-iodoanilino)-N-(2-hydroxyethoxy)-5-[(3-oxoxadinan-2-yl)methyl]benzamide), TAK-733 (3-[(2R)-2,3-dihydroxypropyl]-6-fluoro-5- (2-Fluoro-4-iodoanilino)-8-methylpyrido[2,3-d]pyrimidine-4,7-dione), PD0325901 (N-[(2R)-2,3-dihydroxypropoxy]-3,4-difluoro-2-(2-fluoro-4-iodoanilino)benzamide), CI-1040 (2-(2-chloro-4-iodophenylamino)-N-(cyclopropylmethoxy)-3,4-difluorobenzamide), PD318088 (5-bromo-N-(2,3-dihyd Examples include, but are not limited to, Roxypropoxy)-3,4-difluoro-2-(2-fluoro-4-iodophenylamino)benzamide), PD98059 (2-(2-amino-3-methoxyphenyl)-4H-chromen-4-one), PD334581 (N-[5-[3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]phenyl]-1,3,4-oxadiazole-2-yl]-4-morpholineethanamine), FCN-159, CS3006, HL-085, SHR 7390, and WX-554.

[0130] In one embodiment, the MEK inhibitor is trametinib.

[0131] mTOR inhibitors This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is an mTOR inhibitor.

[0132] Examples of mTOR inhibitors for use in the methods provided herein include everolimus, rapamycin, zotarolimus (ABT-578), ridafololimus (defololimus, MK-8669), sapanicertib, buparlisib, pictilisib, bisutucertib, dactricib, and Torin-1(1-(4-(4-propionylpiperazin-1-yl)-3-(trifluoromethyl)cyclohexyl)-9-(quinoline-3-yl)benzo[h][1,6 Examples include, but are not limited to, naphthyridine-2(1H)-one, GDC-0349((S)-1-ethyl-3-(4-(4-(3-methylmorpholino)-7-(oxetan-3-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidine-2-yl)phenyl)urea, and VS-5584(SB2343,(5-(8-methyl-2-morpholin-4-yl-9-propan-2-ylpurine-6-yl)pyrimidine-2-amine).

[0133] In one embodiment, the mTOR inhibitor is everolimus.

[0134] PD-1 inhibitors This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is a PD-1 inhibitor.

[0135] Examples of PD-1 inhibitors for use in the methods provided herein include, but are not limited to, pembrolizumab, nivolumab, semiprimab, spartalizumab (PDR001), camrelizumab (SHR1210), cintilimab (IBI308), tisrelizumab (BGB-A317), tripalimab (JS001), dostallimab (TSR-042, WBP-285), INCMGA00012 (MGA012), AMP-224, AMP-514, and anti-PD-1 antibodies described in U.S. Patent No. 10,640,504B2 ("Anti-PD-1 Antibody A," paragraph 66, lines 56 to 67, lines 24 and 67, lines 54-57), which are incorporated herein by reference.

[0136] In one embodiment, the PD-1 inhibitor is pembrolizumab. In another embodiment, the PD-1 inhibitor is anti-PD-1 antibody A.

[0137] PD-L1 inhibitors This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is a PD-L1 inhibitor.

[0138] Examples of PD-L1 inhibitors for use in the methods provided herein include, but are not limited to, atezolizumab, avelumab, durvalumab, ZKAB001, TG-1501, SHR-1316, MSB2311, MDX-1105, KN035, IMC-001, HLX20, FAZ053, CS1001, CK-301, CBT-502, BGB-A333, BCD-135, and A167.

[0139] In one embodiment, the PD-L1 inhibitor is atezolizumab.

[0140] PI3K inhibitors This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is a PI3K inhibitor.

[0141] Examples of PI3K inhibitors for use in the methods provided herein include idelalisib, copanlisib, dubelisib, alpelisib, taselicib, perifosin, buparlisib, umbralicib, pictilisib, dactlicib, voxtalisib, sonolisib, tenalisib, serabelisib, acalisib, and CUDC-907(N-hydroxy-2-[[2-(6-methoxypyridine-3-yl)-4-morpholine-4- Iltieno[3,2-d]pyrimidine-6-yl]methyl-methylamino]pyrimidine-5-carboxamide), ME-401(N-[2-methyl-1-[2-(1-methylpiperidine-4-yl)phenyl]propan-2-yl]-4-(2-methylsulfonylbenzimidazole-1-yl)-6-morpholine-4-yl-1,3,5-triazine-2-amine), IPI-549(2-amino-N-[(1S)-1-[8-[2-(1-methylpyrazole-4-yl)ethinyl]-1-oxo- 2-Phenylisoquinoline-3-yl]ethyl]pyrazolo[1,5-a]pyrimidine-3-carboxamide), SF1126((2S)-2-[[(2S)-3-carboxy-2-[[2-[[(2S)-5-(diaminomethylideneamino)-2-[[4-oxo-4-[[4-(4-oxo-8-phenylchromen-2-yl)morpholine-4-ium-4-yl]methoxy]butanoyl]amino]pentanoyl]amino]acetyl]amino]propanoyl]amino]-3-hydroxypropanoyl Examples include, but are not limited to, XL147 (N-[3-(2,1,3-benzothiadiazole-5-ylamino)quinoxaline-2-yl]-4-methylbenzenesulfonamide), GSK1059615 ((5Z)-5-[(4-pyridine-4-ylquinoline-6-yl)methylidene]-1,3-thiazolidined-2,4-dione), and AMG319 (N-[(1S)-1-(7-fluoro-2-pyridine-2-ylquinoline-3-yl)ethyl]-7H-purine-6-amine).

[0142] Raf kinase inhibitors This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is a Raf kinase inhibitor.

[0143] As used herein, the term “RAF kinase” refers to a member of mammalian serine / threonine kinases consisting of three isotypes (C-Raf, B-Raf, and A-Raf), including homodimers of each isotype and heterodimers between isotypes, such as the C-Raf / B-Raf heterodimer.

[0144] As used herein, the term “Raf kinase inhibitor” refers to a compound capable of negatively regulating or inhibiting all or part of the enzymatic activity of one or more members of the Raf family kinases, or of inhibiting activity by disrupting Raf homodimer or heterodimer formation.

[0145] In one embodiment, the Raf kinase inhibitors include encorafenib, sorafenib, lifirafenib, vemurafenib, dabrafenib, PLX-8394 (N-(3-(5-(2-cyclopropylpyrimidine-5-yl)-3a,7a-dihydro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide), and Raf-709 (N-(2-methyl-5,-morpholino-6'- ((Tetrahydro-2H-pyran-4-yl)oxy)-[3,3'-bipyridine]-5-yl)-3-(trifluoromethyl)benzamide), LXH254(N-(3-(2-(2-hydroxyethoxy)-6-morpholinopyridine-4-yl)-4-methylphenyl)-2-(trifluoromethyl)isonicotinamide), LY3009120(1-(3,3-dimethylbutyl)-3-(2-fluoro-4-methyl-5-(7-methyl-2-(methylamino)pyrido[2,3-d]pyrim Zin-6-yl)phenyl)urea), Tak-632(N-(7-cyano-6-(4-fluoro-3-(2-(3-(trifluoromethyl)phenyl)acetamide)phenoxy)benzo[d]thiazole-2-yl)cyclopropanecarboxamide), CEP-32496(1-(3-((6,7-dimethoxyquinazoline-4-yl)oxy)phenyl)-3-(5-(1,1,1-trifluoro-2-methylpropane-2-yl)isoxazole-3-yl)urea), CCT196 Examples include, but are not limited to, 969(1-(3-(tert-butyl)-1-phenyl-1H-pyrazole-5-yl)-3-(2-fluoro-4-((3-oxo-3,4-dihydropyrido[2,3-b]pyrazine-8-yl)oxy)phenyl)urea) and RO5126766(N-[3-fluoro-4-[[4-methyl-2-oxo-7-(2-pyrimidinyloxy)-2H-1-benzopyran-3-yl]methyl]-2-pyridinyl]-N'-methyl-sulfamide).

[0146] In one embodiment, the Raf kinase inhibitor is encorafenib. In one embodiment, the Raf kinase inhibitor is sorafenib. In one embodiment, the Raf kinase inhibitor is rifilafenib.

[0147] SHP2 inhibitors This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is an SHP2 inhibitor.

[0148] Examples of SHP2 inhibitors for use in the methods provided herein include, but are not limited to, SHP-099 (6-(4-amino-4-methylpiperidine-1-yl)-3-(2,3-dichlorophenyl)pyrazine-2-amine dihydrochloride), RMC-4550 ([3-[(3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decane-8-yl]-6-(2,3-dichlorophenyl)-5-methylpyrazine-2-yl]methanol), TNO155, (3S,4S)-8-[6-amino-5-(2-amino-3-chloropyridine-4-yl)sulfanylpyrazine-2-yl]-3-methyl-2-oxa-8-azaspiro[4.5]decane-4-amine), and RMC-4630 (Revolution Medicine). In one embodiment, the SHP inhibitor for use in the method provided herein is RMC-4630 (Revolution Medicine).

[0149] In another embodiment, exemplary SHP2 inhibitors for use in the methods provided herein include 3-[(1R,3R)-1-amino-3-methoxy-8-azaspiro[4.5]deca-8-yl]-6-(2,3-dichlorophenyl)-5-methyl-2-pyrazinemethanol (CAS 2172651-08-8), 3-[(3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]deca-8-yl]-6-[(2,3-dichlorophenyl)thio]-5-methyl-2-pyrazinemethanol (CAS 2172652-13-8), and 3-[(3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]deca-8-yl]-6-[[3-chloro-2-(3-hydroxy Examples include, but are not limited to, 1-azetidinyl)-4-pyridinyl]thio]-5-methyl-2-pyrazinemethanol (CAS 2172652-38-7) and 6-[(2-amino-3-chloro-4-pyridinyl)thio]-3-[(3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]deca-8-yl]-5-methyl-2-pyrazinemethanol (CAS 2172652-48-9).

[0150] In another embodiment, exemplary SHP2 inhibitors for use in the methods provided herein include 1-[5-(2,3-dichlorophenyl)-6-methylimidazo[1,5-a]pyrazine-8-yl]-4-methyl-4-piperidineamine (CAS 2240981-75-1) and (1R)-8-[5-(2,3-dichlorophenyl)-6-methylimidazo[1,5-a]pyrazine-8-yl]-8-azaspiro[4.5]decane-1-amine (CAS S2240981-78-4), (3S,4S)-8-[7-(2,3-dichlorophenyl)-6-methylpyrazolo[1,5-a]pyrazine-4-yl]-3-methyl-2-oxa-8-azaspiro[4.5]decane-4-amine (CAS2240982-45-8), (3S,4S)-8-[7-[(2-amino-3-chloro-4-pyridinyl)thio]pyrazolo[1,5-a]pyrazine-4-yl]-3-methyl-2-oxa-8-azaspiro[4. 5] Decane-4-amine (CAS 2240982-57-2), 4-[(3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]deca-8-yl]-7-(2,3-dichlorophenyl)-6-methyl-pyrazolo[1,5-a]pyrazine-2-methanol (CAS 2240982-69-6), 7-[(2-amino-3-chloro-4-pyridinyl)thio]-4-[(3S,4S)-4-amino-3-methyl-2-oxa-8- Examples include, but are not limited to, azaspiro[4.5]deca-8-yl]-6-methylpyrazolo[1,5-a]pyrazine-2-methanol (CAS 2240982-73-2) and (3S,4S)-8-[7-[(2-amino-3-chloro-4-pyridinyl)thio]-6-methylpyrazolo[1,5-a]pyrazine-4-yl]-3-methyl-2-oxa-8-azspiro[4.5]decane-4-amine (CAS 2240982-77-6).

[0151] In one embodiment, the SHP inhibitor for use in the method provided herein is (1R)-8-[5-(2,3-dichlorophenyl)-6-methylimidazo[1,5-a]pyrazine-8-yl]-8-azaspiro[4.5]decane-1-amine (CAS 2240981-78-4).

[0152] In another embodiment, exemplary SHP2 inhibitors for use in the methods provided herein include 3-[(1R)-1-amino-8-azaspiro[4.5]deca-8-yl]-6-(2,3-dichlorophenyl)-5-hydroxy-2-pyridinemethanol (CAS 2238840-54-3) and 3-[(1R)-1-amino-8-azaspiro[4.5]deca-8-yl]-6-[(2,3-dichlorophenyl)thio]-5-hydroxy-2-pyridinemethanol (CAS 223 (8840-56-5), 5-[(1R)-1-amino-8-azaspiro[4.5]deca-8-yl]-2-(2,3-dichlorophenyl)-3-pyridinol (CAS2238840-58-7), 3-[(1R)-1-amino-8-azaspiro[4.5]deca-8-yl]-6-(2,3-dichlorophenyl)-5-methyl-2-pyridinyl methanol (CAS2238840-60-1), (1R)-8-[6-(2,3-dichlorophenyl)-5-methyl-3-pyridinyl]-8 -Azaspiro[4.5]decane-1-amine (CAS2238840-62-3), 3-[(1R)-1-amino-8-azaspiro[4.5]deca-8-yl]-6-[(2,3-dichlorophenyl)thio]-5-methyl-2-pyridinemethanol (CAS2238840-63-4), (1R)-8-[6-[(2,3-dichlorophenyl)thio]-5-methyl-3-pyridinyl]-8-azaspiro[4.5]decane-1-amine (CAS2238840-64-5), 5-(4 -amino-4-methyl-1-piperidinyl)-2-[(2,3-dichlorophenyl)thio]-3-pyridinol (CAS 2238840-65-6), 5-[(1R)-1-amino-8-azaspiro[4.5]deca-8-yl]-2-[(2,3-dichlorophenyl)thio]-3-pyridinol (CAS 2238840-66-7), 6-[(2-amino-3-chloro-4-pyridinyl)thio]-3-[(3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]deca-8-yl]-5-hydroxy-2-pyridinemethanol (CAS 2238840-67-8), 3-(4-amino-4-methyl-1-piperidinyl)-6-(2,3-dichlorophenyl)-5-hydroxy-2-pyridinemethanol (CAS 2238840-68-9), 3-[(3S,4S)-4-amino-3-methyl-2-oxa-8-azapiro[4.5]deca-8-yl]-6-(2,3-dichlorophenyl)-5-methyl-2-pyridinemethanol (CAS 2238840-69-0), 6-[(2-A [(3S,4S)-4-amino-3-methyl-2-oxaspiro[4.5]deca-8-yl]-5-methyl-2-pyridinemethanol (CAS 2238840-70-3), 3-(4-amino-4-methyl-1-piperidinyl)-6-(2,3-dichlorophenyl)-5-methyl-2-pyridinemethanol (CAS 2238840-71-4), 6-[(2-amino-3-chloro-4-pyridine)thio]-3-(4-amino-4-methyl-1-piperidinyl)- 2-Pyridinemethanol (CAS 2238840-72-5), 5-[(2-amino-3-chloro-4-pyridinyl)thio]-2-[(3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]deca-8-yl]-6-methyl-3-pyridinethanol (CAS 2238840-73-6), 2-[(3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]deca-8-yl]-5-(2,3-dichlorophenyl)-6-methyl-3-pyridinethanol (CAS 223884 Examples include, but are not limited to, 0-74-7), 3-[(3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]deca-8-yl]-6-(2,3-dichlorophenyl)-5-hydroxy-2-pyridinemethanol (CAS 2238840-75-8), and 2-[(2-amino-3-chloro-4-pyridyl)sulfanyl]-5-[(3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl]-6-(hydroxymethyl)pyridine-3-ol.

[0153] In one embodiment, the SHP inhibitor for use in the method provided herein is 3-[(1R)-1-amino-8-azaspiro[4.5]deca-8-yl]-6-[(2,3-dichlorophenyl)thio]-5-hydroxy-2-pyridinemethanol (CAS 2238840-56-5).

[0154] In one embodiment, the SHP2 inhibitors for use in the manner provided herein are the inhibitors disclosed in U.S. Patent Publication No. 10,590,090B2, U.S. Patent Application Publication No. 2020 / 017517A1, U.S. Patent Application Publication No. 2020 / 017511A1, or International Publication No. 2019 / 075265A1, each of which is incorporated herein by whole reference.

[0155] SOS1 inhibitors This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is an SOS1 inhibitor.

[0156] Examples of SOS1 inhibitors for use in the methods provided herein include, but are not limited to, BI3406 (N-[(1R)-1-[3-amino-5-(trifluoromethyl)phenyl]ethyl]-7-methoxy-2-methyl-6-[(3S)-oxolan-3-yl]oxyquinazoline-4-amine) and BI1701963.

[0157] Src kinase inhibitors This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is a Src kinase inhibitor.

[0158] As used herein, the term “Src kinase” refers to members of the mammalian non-receptor tyrosine kinase family, including: Src, Yes, Fyn, and Fgr (SrcA subfamily); Lck, Hck, Blk, and Lyn (SrcB subfamily); and the Frk subfamily.

[0159] As used herein, the term "Src kinase inhibitor" refers to a compound capable of negatively modulating or inhibiting all or part of the enzymatic activity of one or more members of Src kinase.

[0160] Examples of Src kinase inhibitors for use in the methods provided herein include dasatinib, ponatinib, vandetanib, bosutinib, salakatinib, KX2-391 (N-benzyl-2-(5-(4-(2-morpholinoethoxy)phenyl)pyridine-2-yl)acetamide), SU6656 ((Z)-N,N-dimethyl-2-oxo-3-((4,5,6,7-tetrahydro-1H-indole-2-yl)methylene)indoline-5-sulfonamide), PP1 ( Examples include, but are not limited to, 1-(tert-butyl)-3-(p-tolyl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine), WH-4-023(2,6-dimethylphenyl(2,4-dimethoxyphenyl)(2-((4-(4-methylpiperazine-1-yl)phenyl)amino)pyrimidine-4-yl)carbamate), and KX-01(N-benzyl-2-(5-(4-(2-morpholinoethoxy)phenyl)pyridine-2-yl)acetamide).

[0161] In one embodiment, the Src kinase inhibitor is dasatinib. In one embodiment, the Src kinase inhibitor is salakatinib. In one embodiment, the Src kinase inhibitor is ponatinib. In one embodiment, the Src kinase inhibitor is vandetanib. In one embodiment, the Src kinase inhibitor is KX-01.

[0162] Chemotherapy agents This specification provides any one of the methods described in Embodiments 54 to 61, further comprising the simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is one or more chemotherapeutic agents.

[0163] Exemplary chemotherapeutic agents for use in the methods provided herein include, but are not limited to, leucovorin calcium (calcium fophosphate), 5-fluorouracil, irinotecan, oxaliplatin, cisplatin, carboplatin, pemetrexed, docetaxel, paclitaxel, gemcitabine, vinorelbine, chlorambucil, cyclophosphamide, and methotrexate.

[0164] definition The following definitions are provided to facilitate understanding of the scope of this disclosure.

[0165] Unless otherwise indicated, all figures used in this specification or in the claims, representing quantities of components, reaction conditions, etc., should be understood in all cases to be modified by the term “approximately.” Therefore, unless otherwise indicated, the numerical parameters described in the following specification and the attached claims are approximations that may vary depending on the standard deviation found in each of their test measurements.

[0166] When used herein, if any variable element is present in two or more instances in a chemical formula, its definition in each instance is independent of its definition in all other instances. If the chemical structure and chemical name conflict, the identity of the compound is determined by the chemical structure.

[0167] stereoisomer The compounds of this disclosure may contain, for example, double bonds, one or more chiral carbon atoms, and rotationally hindered bonds, and therefore may exist as stereoisomers such as double bond isomers (i.e., geometric isomers (E / Z)), enantiomers, diastereomers, and atropisomers. Accordingly, it should be understood that the scope of this disclosure includes all possible stereoisomers of the exemplary compounds, including stereoisomerically pure forms (e.g., geometrically pure, enantiomerically pure, diastereoisomerically pure, and atropisomerically pure), as well as mixtures of stereoisomers of any chemical structure (all or part) disclosed herein (e.g., mixtures of geometric isomers, enantiomers, diastereomers, and atropisomers, or any of the aforementioned mixtures), unless the stereochemistry is specifically identified.

[0168] If the stereochemistry of a structure or a part of a structure is not indicated by, for example, a thick line or a dashed line, this structure or part of a structure should be interpreted as encompassing all of its stereoisomers. If the stereochemistry of a structure or a part of a structure is not indicated by, for example, a thick line or a dashed line, unless otherwise specified, this structure or part of a structure should be interpreted as encompassing only the specified stereoisomer.

[0169] for example, [ka] teeth, [ka] Similarly, for example, the chemical name (4R)-4-methoxy-5-methyl-4,5,6,7-tetrahydro-2H-isoindole represents (4R,5R)-4-methoxy-5-methyl-4,5,6,7-tetrahydro-2H-isoindole and (4R,5S)-4-methoxy-5-methyl-4,5,6,7-tetrahydro-2H-isoindole.

[0170] As a further example, [ka] teeth, [ka] Similarly, for example, the chemical name 7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridine-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione represents (M)-7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridine-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione and (P)-7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridine-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione.

[0171] In certain cases, a dashed line may be used to indicate that both stereoisomers are contained within the bond. This dashed line should not be confused with a dashed line drawn perpendicular to the bond (indicating the point where the group is attached to the rest of the molecule).

[0172] As used herein, the terms “stereoisomer” or “stereoisomerically pure” mean a single stereoisomer of a compound (e.g., geometric isomer, enantiomer, diastereomer, and atropisomer) that substantially does not contain any other stereoisomers of the compound. For example, a stereoisomerically pure compound having one chiral center substantially does not contain any enantiomers of the compound, and a stereoisomerically pure compound having two chiral centers substantially does not contain any other enantiomers or diastereomers of the compound. A typical stereoisomerically pure compound includes more than 80% by weight of one stereoisomer of the compound and less than 20% by weight of other stereoisomers of the compound, more than 90% by weight of one stereoisomer of the compound and less than 10% by weight of other stereoisomers of the compound, more than 95% by weight of one stereoisomer of the compound and less than 5% by weight of other stereoisomers of the compound, or more than 97% by weight of one stereoisomer of the compound and less than 3% by weight of other stereoisomers of the compound.

[0173] This disclosure also includes pharmaceutical compositions containing stereoisomerically pure forms and the use of stereoisomerically pure forms of any compound disclosed herein. Furthermore, this disclosure also includes pharmaceutical compositions containing mixtures of stereoisomers of any compound disclosed herein, and the use of the above-mentioned pharmaceutical compositions or mixtures of stereoisomers. These stereoisomers or mixtures thereof can be synthesized according to methods well known in the art and methods disclosed herein. Mixtures of stereoisomers can be resolved using standard methods such as chiral columns or chiral resolving agents. For example, see Jacques et al., Enantiomers, Racemates and Resolutions (Wiley-Interscience, New York, 1981); Wilen et al., Tetrahedron 33:2725; Eliel, Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, Tables of Resolving Agents and Optical Resolutions, page 268 (Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN, 1972).

[0174] Tautomers As is known to those skilled in the art, certain compounds disclosed herein may exist in one or more tautomerized forms. Since one chemical structure may be used to represent only one tautomerized form, for convenience, a reference to a compound of a given structural formula should be understood to include other tautomers of that structural formula. For example, [ka] teeth, [ka] Similarly, for example, the chemical name (4R,5R)-4-methoxy-5-methyl-4,5,6,7-tetrahydro-1H-indazole represents (4R,5R)-4-methoxy-5-methyl-4,5,6,7-tetrahydro-1H-indazole and (4R,5R)-4-methoxy-5-methyl-4,5,6,7-tetrahydro-2H-indazole.

[0175] Therefore, the scope of this disclosure should be understood to encompass all tautomeristic forms of the compounds disclosed herein.

[0176] isotope labeled compounds Furthermore, the scope of this disclosure includes all pharmaceutically acceptable isotope-labeled compounds of the compounds disclosed herein, such as the compounds of Formula I, where one or more atoms are substituted by atoms having the same atomic number but with an atomic mass or mass number different from those commonly found in nature. Examples of isotopes suitable for inclusion in the compounds disclosed herein include: 2 H and 3 Hydrogen such as H, 11 C, 13 C and 14 Carbon such as C, 36 Chlorine such as Cl 18 Fluorine such as F 123 I and 125 Iodine such as I 13 N and 15 Nitrogen such as N, 15 O, 17 O and 18 Oxygen such as O, 32 Phosphorus such as P, and 35 Examples include sulfur isotopes such as ₂S. Certain isotope-labeled compounds of formula I, for example, those incorporating radioactive isotopes, are useful for studying the tissue distribution of drugs and / or substrates. Radioactive isotope tritium ( 3 H) and carbon-14 ( 14 C) is particularly useful for this purpose, considering that it is easy to incorporate and is an easy detection means. Deuterium ( 2Substitution with isotopes such as H or D may result in certain therapeutic benefits, such as increased in vivo half-life or reduced dosage requirements, which may be advantageous in some cases, as they lead to greater metabolic stability. 11 C, 18 F, 15 O and 13 Substitution with positron-emitting isotopes such as 1N may be useful, for example, in positron emission tomography (PET) studies to investigate target occupancy. The isotopically labeled compounds of the compounds disclosed herein can usually be prepared by conventional methods known to those skilled in the art, or by processes similar to those described in the accompanying general synthesis procedures and examples, using appropriate isotopically labeled reagents instead of conventionally used unlabeled reagents.

[0177] Various definitions This section defines additional terms used to describe the compounds, compositions, and scope of use disclosed herein.

[0178] The terms "2h-binding exchange assay" or "20h-binding exchange assay," as used herein, refer to the assay described in the section titled "Biological Evaluation."

[0179] When used in this specification, the term "C 1~3 Alkyl," "C 1~4 "Alkyl" and "C 1~6 "Alkyl" refers to a straight-chain or branched-chain hydrocarbon containing 1 to 3, 1 to 4, and 1 to 6 carbon atoms, respectively. 1~3 Alkyl, C 1~4 Alkyl, or C 1~6 Typical examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, pentyl, and hexyl.

[0180] When used in this specification, the term "C 1~3 "alkoxy" and "C 1~4 "alkoxy" is -OR #This refers to R as defined herein. # These are C 1~4 Alkyl and C 1~4 Represents an alkyl group. C 1~3 Alkoxy or C 1~4 Typical examples of alkoxys include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, and butoxy.

[0181] When used in this specification, the term "C 3~5 "Cycloalkyl" and "C 3~6 "Cycloalkyl" refers to saturated carbocyclic molecules whose cyclic skeletons have 3 to 5 and 3 to 6 carbon atoms, respectively. 3~5 Cycloalkyl or C 3~6 Typical examples of cycloalkyl compounds include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0182] When used in this specification, the term "C 1~4 "Dialkylamino" refers to -NR*R**, where R* and R** are independently defined as C as defined herein. 1~4 Represents alkyl. 1~4 Typical examples of dialkylaminos include, but are not limited to, -N(CH3)2, -N(CH2CH3)2, -N(CH3)(CH2CH3), -N(CH2CH2CH3)2, and -N(CH(CH3)2)2.

[0183] When used in this specification, the term "C 1~4 "Alkylamino" refers to -NHR*, where R* is C as defined herein. 1~4 Represents alkyl. 1~4 Typical examples of alkylaminos include, but are not limited to, -NH(CH3), -NH(CH2CH3), -NH(CH2CH2CH3), and -NH(CH(CH3)2).

[0184] As used herein, the term "halogen" refers to -F, -CI, -Br, or -I.

[0185] When used herein as a prefix to other terms relating to chemical groups, the term "halo" refers to a modification of a chemical group in which one or more hydrogen atoms are replaced by one or more halogen atoms as defined herein. The halogens are selected independently for each occurrence. For example, the term "C 1~4 "Haloalkyl" is defined herein as a C atom in which one or more hydrogen atoms are substituted with halogens. 1~4 Refers to alkyl. 1~4 Typical examples of haloalkyls include, but are not limited to, -CH2F, -CHF2, -CF3, -CHFCl, -CH2CF3, -CFHCF3, -CF2CF3, -CH(CF3)2, -CF(CHF2)2, and -CH(CH2F)(CF3). Furthermore, for example, the term "C 1~4 "Haloalkoxy" is, for example, a C as defined herein, in which one or more hydrogen atoms are substituted with halogens. 1~4 Refers to alkoxy. 1~4 Typical examples of haloalkoxys include, but are not limited to, -OCH2F, -OCHF2, -OCF3, -OCHFCl, -OCH2CF3, -OCFHCF3, -OCF2CF3, -OCH(CF3)2, -OCF(CHF2)2, and -OCH(CH2F)(CF3).

[0186] As used herein, the terms “5-6 membered heteroaryl” and “5-10 membered heteroaryl” refer to monocyclic or bicyclic aromatic ring systems containing 1-5 and 1-10 heteroatoms, respectively, independently selected from N, O, and S, with the remaining ring atoms being carbon. Typical examples of 5-6 or 5-10 membered heteroaryls include furanyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazolyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl, triazolyl, pyridyl, pyridadinyl, pyrazinyl, pyrimidinyl, benzofuranyl, benzimidazolyl, benzoisoxazolyl, benzopyranyl, benzothiadiazolyl, benzothiazolyl, benzothienyl, benzothiophenyl, and benz. Examples include, but are not limited to, zotriazolyl, benzoxazolyl, phlopyridyl, imidazopyridinyl, imidazothiazolyl, indolidinyl, indazolyl, isobenzofuranyl, isobenzothienyl, isoindolyl, isoquinolinyl, isothiazolyl, naphthilidinyl, oxazolopyridinyl, phthalazinyl, pteridinyl, purinyl, pyridopyridyl, pyrrolopyridyl, quinolinyl, quinoxalinyl, chiazolinyl, thiadiazolopyridyl, and thienopyridyl.

[0187] When used in this specification, the term "C 3~5 "heterocycloalkyl" and "C 3~6 A "heterocycloalkyl" refers to a saturated carbocyclic molecule in which the cyclic skeleton has 3 to 5 and 3 to 6 carbon atoms respectively, and one or two carbon atoms are independently substituted with one or two heteroatoms selected from N, O, and S. 3~5 Heterocycloalkyl or C 3~6 Typical examples of heterocycloalkyls include, but are not limited to, azilidinyl, azetidinyl, oxetanyl, pyrrolidinyl, piperidinyl, piperazinyl, or morpholinyl.

[0188] As used herein, the term “pharmaceutically acceptable” generally refers to something that is recognized for use in a subject, particularly in humans.

[0189] As used herein, the term “pharmaceutically acceptable salt” refers to a salt of a compound that is pharmaceutically acceptable and has the desired pharmacological activity of the parent compound. Such salts include (1) acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid; or acid addition salts formed with organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, and methanesulfonic acid; or (2) salts formed when the acidic proton present in the parent compound is replaced by a metal ion, such as an alkali metal ion, an alkaline earth ion, or an aluminum ion; or salts formed when the compound is coordinated with an organic base such as ethanolamine, diethanolamine, triethanolamine, N-methylglucamine, or dicyclohexylamine. Further examples of such salts can be found in Berge et al., J. Pharm. Sci. 66(1):1-19 (1977). Also, see Stahl et al., Pharmaceutical Salts: Properties, Selection, and Use, 2 nd See also the Revised Edition (2011).

[0190] As used herein, the term “pharmaceutically acceptable excipient” refers to a wide range of components that can be combined with the compounds or salts disclosed herein to prepare a pharmaceutical composition or formulation. Typical excipients include, but are not limited to, diluents, colorants, vehicles, anti-tacks, flow enhancers, disintegrants, flavorings, coatings, binders, sweeteners, lubricants, adsorbents, and preservatives.

[0191] As used herein, the term “subject” refers to humans and mammals, including but not limited to primates, cattle, sheep, goats, horses, dogs, cats, rabbits, rats, and mice. In one embodiment, the subject is a human.

[0192] As used herein, the term “therapeutic dose” means the amount of a compound disclosed herein that elicits a biological or medical response in a tissue, system, or subject, as determined by a researcher, veterinarian, doctor of medicine, or other clinician.

[0193] General synthesis procedure The compounds provided herein can be synthesized according to the procedures described in this section and the following sections. The synthetic methods described herein are merely illustrative, and the compounds disclosed herein can also be synthesized by alternative routes using alternative synthetic strategies as understood by those skilled in the art. It should be understood that the general synthetic procedures and specific examples provided herein are illustrative and should not be construed as limiting the scope of this disclosure.

[0194] In general, compounds of formula I can be synthesized according to the following scheme. Any variable elements used in the following scheme are the same as those defined for formula I, unless otherwise noted. All starting materials can be commercially available, for example, from Sigma-Aldrich, Inc., or known in the art, or synthesized using known procedures employing conventional techniques. Starting materials can also be synthesized via the procedures disclosed herein. Preferred reaction conditions, such as solvents, reaction temperatures, and reagents, for the schemes described in this section can be found in the examples provided herein. [ka]

[0195] As will be understood by those skilled in the art, the above-described synthesis schemes and representative examples are not intended to be a comprehensive list of all means by which the compounds described and claimed in this application can be synthesized. Further methods will become apparent to those skilled in the art. In addition, the various synthesis steps described above can be carried out in alternative orders or sequences to obtain the desired compounds.

[0196] The methods for purifying the compounds described herein are known in the art and include, for example, crystallization, chromatography (e.g., liquid phase and gas phase), extraction, distillation, tritulation, and reverse-phase HPLC.

[0197] This disclosure further includes “intermediate” compounds, which include structures produced by the described synthetic procedure before obtaining the final desired compound, whether isolated or generated in situ and not isolated. These intermediates are within the scope of this disclosure. Exemplary embodiments of such intermediate compounds are shown in the following examples. [Examples]

[0198] This section provides specific examples of compounds of formula I and methods for producing them.

[0199] List of abbreviations

[0200] [Table 2]

[0201] [Table 3]

[0202] General analytical and purification methods This section provides a description of general analytical and purification methods used to prepare the specific examples provided herein.

[0203] Chromatography: Unless otherwise specified, residues containing the crude product were purified by passing the crude material or concentrate through a pre-packed silica gel column of either Biotage or ISCO brand, either flash silica (SiO2) or reversed-phase flash silica (C18), and eluting the product from the column using the specified solvent gradient. For example, the description (330 g SiO2, 0-40% HCl / hexane) means that the product was obtained by eluting a column packed with 330 grams of silica with a solvent gradient of 0-40% HCl in hexane.

[0204] Preparative HPLC method: Where indicated, the compounds described herein were purified by reverse-phase HPLC using a Waters FractionLynx semi-preparative HPLC-MS system with one of the following two HPLC columns: (a) a Phenomenex Gemini column (5 micron, C18, 150 × 30 mm) or (b) a Waters X-select CSH column (5 micron, C18, 100 × 30 mm).

[0205] A typical instrumental experiment involves eluting 10% (v / v) to 100% MeCN (0.1% v / v formic acid) in water (0.1% formic acid) at a rate of 45 mL / min over 10 minutes, with conditions that may be varied to achieve optimal separation.

[0206] Preparative SFC method: When indicated in this manner, the compounds described herein were purified via SFC using Chiral SFC-80 (Thar,Waters) in an AD (20 × 250 mm, 10 μm) (Daicel) column.

[0207] Proton NMR spectrum: Unless otherwise instructed, all 1¹H NMR spectra were collected at 300, 400, or 500 MHz using a Bruker NMR instrument. When characterized in this manner, all observed protons are reported as parts per million (ppm) of low magnetic field from tetramethylsilane (TMS) using the internal solvent peak as a reference.

[0208] Mass spectrum (MS) Unless otherwise indicated, all mass spectral data for the starting material, intermediates, and / or exemplary compounds are given in [M+H] format. + The molecular ion is reported as mass / charge (m / z). The reported molecular ions were obtained by electrospray detection (commonly referred to as ESI MS) using a Waters Acquity UPLC / MS system. As will be understood by those skilled in the art, compounds with isotopic atoms, such as bromine, are usually reported according to the detected isotopic pattern.

[0209] Name of compound The compounds disclosed and described herein have been named using the IUPAC naming functionality provided by JChem for Excel 18.22.1.7 from ChemAxon Ltd.

[0210] Specific Examples This section provides procedures for synthesizing specific examples of the compounds provided herein. Unless otherwise noted, all starting materials are commercially available from Merck Sigma-Aldrich Inc. or are known in the art and can be synthesized using known procedures employing the usual art.

[0211] Synthesis of the Examples Method 1 Example 1-1: (S)-3-((2-(2-acryloyl-2,6-diazaspiro[3,4]octan-6-yl)-5,6,7,8-tetrahydroquinazoline-4-yl)amino)-N,5-dimethylhexaneamide [ka] Step 1: (S)-3-((2-chloro-5,6,7,8-tetrahydroquinazolin-4-yl)amino)-N,5-dimethylhexaneamide(A-1) A mixture of 2,4-dichloro-5,6,7,8-tetrahydroquinazoline (0.21 g, 1.01 mmol, Combi-Blocks), (S)-3-amino-N,5-dimethylhexaneamide dihydrochloride (0.31 g, 1.31 mmol, Angel Pharmatech), DIPEA (1.06 mL, 6.06 mmol, Aldrich), and DMA (4 mL) was heated at 100°C for 17 hours. After completion, the reaction mixture was washed with saturated NH4Cl and water and extracted with SiO2. The combined organic extract was dried over Na2SO4, filtered, and concentrated to obtain (S)-3-((2-chloro-5,6,7,8-tetrahydroquinazoline-4-yl)amino)-N,5-dimethylhexaneamide A-1 (yield obtained over two steps) as a pale yellow oil, which was used directly in the next step. m / z(ESI):325.2(M+H) + .

[0212] Step 2: tert-butyl(S)-6-(4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3,4]octane-2-carboxylate(B-1) A mixture of (S)-3-((2-chloro-5,6,7,8-tetrahydroquinazolin-4-yl)amino)-N,5-dimethylhexaneamide A-1 (0.20 g, 0.62 mmol), tert-butyl 2,6-diazaspiro[3.4]octane-2-carboxylate (0.26 g, 1.23 mmol, Combi-Blocks), DIPEA (0.32 mL, 1.85 mmol, Aldrich), and DMA (4 mL) was heated at 130°C for 7 hours. After completion, the reaction mixture was washed with saturated NH4Cl and water and extracted with siRNA. The combined organic extracts were concentrated and purified by silica gel chromatography using 0-40% (3:1 siRNA / EtOH) in heptane to obtain tert-butyl(S)-6-(4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate B-1 (0.25 g, 0.62 mmol, yield 100%) as a yellow oil. m / z(ESI): 501.2(M+H) + .

[0213] Step 3: (S)-3-((2-(2,6-diazaspiro[3,4]octan-6-yl)-5,6,7,8-tetrahydroquinazoline-4-yl)amino)-N,5-dimethylhexaneamide(C-1) To a solution of tert-butyl(S)-6-(4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate B-1 (0.25 g, 0.62 mmol) in DCM (2 mL), TFA (4.0 mL, 51.9 mmol, Aldrich) was added. The resulting mixture was stirred at rt for 30 minutes. After completion, the reaction product was concentrated, washed with 10% Na2CO3, and extracted with DCM and 3:1 (siRNA / EtOH). The combined organic extract was dried with Na2SO4, filtered, and concentrated to obtain (S)-3-((2-(2,6-diazaspiro[3,4]octan-6-yl)-5,6,7,8-tetrahydroquinazoline-4-yl)amino)-N,5-dimethylhexaneamide C-1 (yield obtained over two steps) as a white solid, which was used directly in the next step. m / z(ESI):401.2(M+H) + .

[0214] Step 4: (S)-3-((2-(2-acryloyl-2,6-diazaspiro[3,4]octan-6-yl)-5,6,7,8-tetrahydroquinazoline-4-yl)amino)-N,5-dimethylhexaneamide (Example 1-1) To a solution of (S)-3-((2-(2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydroquinazolin-4-yl)amino)-N,5-dimethylhexaneamide C-1 (0.20 g, 0.50 mmol) in DCM (10 mL), acryloyl chloride (2.50 mL, 0.50 mmol, Aldrich) was added. The solution was stirred at rt for 30 minutes. The reaction product was washed with saturated NaHCO3 and extracted with DCM and 3:1 (siRNA / EtOH). The combined organic extracts were dried over Na2SO4, filtered, concentrated, and chromatographed on silica gel using 0-10% MeOH in DCM to obtain (S)-3-((2-(2-acryloyl-2,6-diazaspiro[3,4]octan-6-yl)-5,6,7,8-tetrahydroquinazoline-4-yl)amino)-N,5-dimethylhexaneamide Example 1-1 (0.116 g, 0.255 mmol, yield 51%) as a white solid. m / z(ESI):455.2(M+H) + . 1 H NMR(400MHz,DMSO-d6)δ ppm 7.79(br d,J=4.0Hz,1H),6.26-6.36(m,1H),6.11(dd,J=16.9,2.3Hz,1H),5.67(dd,J=10.2,2.3Hz,1H),4.52-4.64(m,1H),4.11-4.21( m,2H),3.87(s,2H),3.40-3.70(m,4H),2.55(d,J=4.6Hz,3H),2.40-2.46(m,3H),2.22-2.39(m,3H),2.08-2.22(m,4H),1.69(br d,J=5.6Hz,4H),1.51-1.64(m,2H),0.87(dd,J=6.3,3.6Hz,6H).

[0215] [Table 4]

[0216] [Table 5]

[0217] Table 6

[0218] Table 7

[0219] Table 8

[0220] Table 9

[0221] Table 10

[0222] Table 11

[0223] Table 12

[0224] Table 13

[0225] Table 14

[0226] Table 15

[0227] Table 16

[0228] [Table 17]

[0229] [Table 18]

[0230] [Table 19]

[0231] [Table 20]

[0232] [Table 21]

[0233] Additional steps 3a and 3b of Example 1-12. [ka] Step 3a: tert-butyl(S)-6-(4-((1-((1-amino-2-methyl-1-oxopropan-2-yl)amino)-5-methyl-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3,4]octane-2-carboxylate A mixture of (R)-3-((2-(2-(tert-butoxycarbonyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydroquinazoline-4-yl)amino)-5-methylhexanoic acid (0.40 g, 0.82 mmol, step 3 of Examples 1-22), 2-amino-2-methylpropanamide hydrochloride (0.114 g, 0.82 mmol, synthesized according to the procedure described in International Publication No. 2008017691), HATU (0.624 g, 1.64 mmol, Spectrochem), and DIPEA (0.43 mL, 2.46 mmol) in DMF (10 mL) was stirred for 16 hours. The reaction mixture was then diluted with ice-cold water, and the precipitated solid was filtered and dried to obtain tert-butyl(S)-6-(4-((1-((1-amino-2-methyl-1-oxopropan-2-yl)amino)-5-methyl-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.40 g, 0.70 mmol, yield 85%) as a grayish-white solid, which was used in the next step without purification. m / z(ESI): 571.8(M+H) + .

[0234] Step 3b: tert-butyl(S)-6-(4-((1-(4,4-dimethyl-5-oxo-4,5-dihydro-1H-imidazole-2-yl)-4-methylpentan-2-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3,4]octane-2-carboxylate To a solution of tert-butyl(S)-6-(4-((1-((1-amino-2-methyl-1-oxopropan-2-yl)amino)-5-methyl-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.35 g, 0.61 mmol) in MeOH (5 mL) and water (5 mL), NaOH (0.61 mL, 1 M) was added, and the resulting mixture was stirred at 100 °C for 16 hours. Subsequently, the reaction mixture was concentrated under reduced pressure, the residue was diluted with water, and extracted by DCM. The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain tert-butyl(S)-6-(4-((1-(4,4-dimethyl-5-oxo-4,5-dihydro-1H-imidazole-2-yl)-4-methylpentan-2-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.30 g, 0.54 mmol, yield 89%) as a grayish-white solid, which was used in the next step without purification. m / z(ESI): 554.0(M+H) + .

[0235] Additional steps 3a and 3b of Examples 1-20. [ka] Step 3a: tert-butyl(S)-6-(4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)-7-(propa-1-en-2-yl)quinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate To a degassed solution of tert-butyl(S)-6-(7-bromo-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)quinazolin-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.30 g, 0.52 mmol), 4,4,5,5-tetramethyl-2-(propa-1-en-2-yl)-1,3,2-dioxaborolane (0.175 g, 1.04 mmol, Chempure), and Na2CO3 (0.166 g, 1.56 mmol), PdCl2(dppf)-DCM adduct (0.043 g, 0.052 mmol, Chempure) was added, and the resulting mixture was heated at 90°C for 16 hours. Next, the reaction mixture was diluted with water and extracted by DCM. The organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by eluting a Redi-Sep pre-packed silica gel column (12 g) with a gradient of 5-10% MeOH in DCM to obtain tert-butyl(S)-6-(4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)-7-(propa-1-en-2-yl)quinazolin-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.20 g, 0.37 mmol, yield 72%) as a brown solid. m / z(ESI): 536.8(M+H) + .

[0236] Step 3b: tert-butyl(S)-6-(7-isopropyl-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)quinazoline-2-yl)-2,6-diazaspiro[3,4]octane-2-carboxylate To a solution of tert-butyl(S)-6-(4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)-7-(propa-1-en-2-yl)quinazolin-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.20 g, 0.373 mmol) in MeOH (2 mL), Pd / C (10%) (0.040 g, 0.037 mmol, Chempure) was added, and the resulting mixture was stirred under an H2 atmosphere for 16 hours. Next, the reaction mixture was filtered through a Celite pad, and the filtrate was concentrated under reduced pressure to obtain tert-butyl(S)-6-(7-isopropyl-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)quinazolin-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.16 g, 0.297 mmol, 80% yield) as a brown solid, which was then transferred to the next step without purification. m / z(ESI):539.0(M+H) + .

[0237] Additional step 5 for Examples 1-21-1 and 1-21-2. [ka] Step 5: tert-butyl6-(4-((1-(3-cyanophenyl)-3-(methylamino)-3-oxopropyl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate A mixture of tert-butyl 6-(4-((1-(3-cyanophenyl)-3-methoxy-3-oxopropyl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (480 mg, 0.878 mmol) and methanamine (33% in EtOH) (10 mL, 0.878 mmol, Spectrochem) was stirred in a sealed tube of rt for 16 hours. The reaction mixture was then concentrated under reduced pressure, and the residue was purified using a Redi-Sep pre-packed silica gel column (12 g) eluted with 5-15% MeOH in DCM to obtain tert-butyl 6-(4-((1-(3-cyanophenyl)-3-(methylamino)-3-oxopropyl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (400 mg, 0.733 mmol, yield 83%) as a rubbery solid. m / z(ESI): 545.8(M+H) + The racemic mixture was separated by chiral SFC in a (S,S)Whelk-01 (250×50mm, 5μ) column using 0.5% diethylamine (1:1) in liquid CO2:MeOH, and tert-butyl(R)-6-(4-((1-(3-cyanophenyl)-3-(methylamino)-3-oxopropyl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3. 4]Octane-2-carboxylate (125 mg, 0.23 mmol, yield 31%) and tert-butyl(S)-6-(4-((1-(3-cyanophenyl)-3-(methylamino)-3-oxopropyl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (110 mg, 0.20 mmol, yield 27%) were obtained as peak 2. The stereochemistry of the structures was arbitrarily assigned and is not determined. Peak 1: 1H NMR(400MHz,DMSO-d6):δ ppm 7.80-7.86(m,2H),7.58-7.70(m,2H),7.48-7.52(m,1H),6.84-6.90(m ,1H),5.41(m,1H),3.65-3.82(m,2H),3.49-3.53(m,2H),3.14-3.30(m ,4H),2.59-2.74(m,4H),2.28-2.45(m,4H),2.10-2.28(m,2H),2.00(t ,J=6.8Hz,2H),1.63-1.74(m,2H),1.37(m,9H).m / z(ESI):545.8(M+H) + Peak 2: 1 H NMR(400MHz,DMSO-d6):δ ppm 7.83(m,2H),7.61-7.71(m,2H),7.48(m,1H),6.87(m,1H),5.36-5.49(m,1H),3.71(s,4H),3.53-3.55(m,2H),3.13-3.27(m,2H), 2.59-2.79(m,4H),2.37(s,4H),2.12-2.32(m,2H),2.00(t,J=6.8Hz,2H),1.64-1.78(m,2H),1.37(m,9H).m / z(ESI):545.8(M+H) + .

[0238] Additional steps 5-7 of Example 1-22. [ka] Step 5: (S)-3-((2-(2-(tert-butoxycarbonyl)-2,6-diazaspiro[3,4]octan-6-yl)-5,6,7,8-tetrahydroquinazoline-4-yl)amino)-5-methylhexanoic acid To a solution of tert-butyl(S)-6-(4-((1-methoxy-5-methyl-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazolin-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (6.7 g, 13.4 mmol) in THF (20 mL), MeOH (10 mL), and water (5 mL), LiOH (1.6 g, 66.8 mmol) was added, and the mixture was stirred at rt for 12 hours. Subsequently, the reaction mixture was concentrated under reduced pressure, the resulting solid was dissolved in water, acidified with 1N HCl, and extracted with SiO2. The combined organic extracts were dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain (S)-3-((2-(2-(tert-butoxycarbonyl)-2,6-diazaspiro[3,4]octan-6-yl)-5,6,7,8-tetrahydroquinazoline-4-yl)amino)-5-methylhexanoic acid (5.2 g, 10.7 mmol, yield 80%) as a grayish-white solid. 1 H NMR(400MHz,DMSO-d6):δ ppm 4.66(s,1H),3.68-3.86(m,5H),3.54-3.64(m,2H),3.45(d,J=10.8Hz,2H),2.33-2.46(m,3H),2.17-2.26 (m,2H),2.02-2.14(m,2H),1.49-1.78(m,7H),1.39(d,J=1.6Hz,10H),1.26(m,1H),0.88(d,J=6.2Hz,6H).

[0239] Step 6: tert-butyl(S)-6-(4-((1-(2-acetylhydrazineyl)-5-methyl-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate A mixture of (S)-3-((2-(2-(tert-butoxycarbonyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydroquinazoline-4-yl)amino)-5-methylhexanoic acid (0.40 g, 0.82 mmol), EDC (0.19 g, 0.98 mmol), HOBt (0.15 g, 0.98 mmol), DIPEA (0.29 mL, 1.64 mmol), and acetohydrazide (0.073 g, 0.98 mmol) in DMF (4 mL) was stirred at rt for 16 hours. The reaction mixture was diluted with ice-cold water and extracted with SiO2. The combined organic extracts were washed with brine, separated, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain tert-butyl(S)-6-(4-((1-(2-acetylhydrazineyl)-5-methyl-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.35 g) as a grayish-white solid, which was then transferred to the next step without purification. m / z(ESI): 543.9(M+H) + .

[0240] Step 7: 1-(6-(4-(((2S)-4-methyl-1-(5-methyl-1,3,4-oxadiazole-2-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3,4]octan-2-yl)-2-propen-1-one A solution of tert-butyl(S)-6-(4-((1-(2-acetylhydrazineyl)-5-methyl-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.35 g, 0.64 mmol) in POCl3 (0.60 mL, 6.4 mmol) was heated at 100 °C for 4 hours. The reaction mixture was concentrated under reduced pressure, the residue was diluted with 1N NaOH solution, and extracted with 10% MeOH in DCM. The combined organic extracts were washed with brine, separated, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain (S)-N-(4-methyl-1-(5-methyl-1,3,4-oxadiazole-2-yl)pentan-2-yl)-2-(2,6-diazaspiro[3,4]octan-6-yl)-5,6,7,8-tetrahydroquinazoline-4-amine, which was then transferred to the next step without purification. m / z(ESI):426.0(M+H) + .

[0241] To a solution of (S)-N-(4-methyl-1-(5-methyl-1,3,4-oxadiazole-2-yl)pentan-2-yl)-2-(2,6-diazaspiro[3,4]octan-6-yl)-5,6,7,8-tetrahydroquinazoline-4-amine (50 mg, 0.068 mmol) in DCM (2 mL), Et3N (28.5 μL, 0.204 mmol) and acryloyl chloride (5.5 μL, 0.068 mmol) were added at -30°C. The reaction mixture was stirred at the same temperature for 30 minutes, then diluted with water and extracted with DCM. The organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by reverse-phase HPLC (Phenomenex Gemini C18 column, 150 × 30 mm, 0.1% TFA in 10-100% MeCN / H2O) to obtain (S)-1-(6-(4-((4-methyl-1-(5-methyl-1,3,4-oxadiazole-2-yl)pentan-2-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octan-2-yl)propa-2-en-1-one (25 mg, 0.052 mmol, yield 76%) as a white solid. 1H NMR(400MHz,DMSO-d6):δ ppm 6.32(dd,J=17.0,10.3Hz,1H),6.11(dd,J=17.0,2.3Hz,1H),5.98(d,J=8.8Hz,1H),5.67(dd,J= 10.2,2.3Hz,1H),4.64-4.67(m,1H),4.12-4.18(m,2H),3.86(d,J=3.9Hz,2H),3.52-3.57(m,2H) ),3.40-3.44(m,2H),2.96-3.02(m,2H),2.32-2.41(m,5H),2.19(d,J=6.9Hz,2H),2.01-2.13(m ,2H),1.62-1.67(m,6H),1.25-1.29(m,1H),0.86(dd,J=15.6,6.4Hz,6H).m / z(ESI):480.2(M+H) + .

[0242] Additional steps 3-6 prior to N-deprotection / acylation in Examples 1-23. [ka] Step 3: (S)-3-((2-(2-(tert-butoxycarbonyl)-2,6-diazaspiro[3,4]octan-6-yl)-5,6,7,8-tetrahydroquinazoline-4-yl)amino)-5-methylhexanoic acid To a solution of tert-butyl(S)-6-(4-((1-methoxy-5-methyl-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazolin-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (6.7 g, 13.4 mmol) in THF (20 mL), MeOH (10 mL), and water (5 mL), LiOH (1.6 g, 66.8 mmol) was added, and the mixture was stirred at rt for 12 hours. Subsequently, the reaction mixture was concentrated under reduced pressure, the resulting solid was dissolved in water, acidified with 1N HCl, and extracted with SiO2. The combined organic extracts were dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain (S)-3-((2-(2-(tert-butoxycarbonyl)-2,6-diazaspiro[3,4]octan-6-yl)-5,6,7,8-tetrahydroquinazoline-4-yl)amino)-5-methylhexanoic acid (5.2 g, 10.7 mmol, yield 80%) as a grayish-white solid. 1 H NMR(400MHz,DMSO-d6):δ ppm 4.66(s,1H),3.68-3.86(m,5H),3.54-3.64(m,2H),3.45(d,J=10.8Hz,2H),2.33-2.46(m,3H),2.17-2.26 (m,2H),2.02-2.14(m,2H),1.49-1.78(m,7H),1.39(d,J=1.6Hz,10H),1.26(m,1H),0.88(d,J=6.2Hz,6H).

[0243] Step 4: tert-butyl(S)-6-(4-((1-hydroxy-5-methylhexane-3-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3,4]octane-2-carboxylate To a solution of (S)-3-((2-(2-(tert-butoxycarbonyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydroquinazoline-4-yl)amino)-5-methylhexanoic acid (2.0 g, 4.1 mmol) in THF (20 mL), borane-THF complex (12.3 mL, 12.3 mmol, 1 M in THF, Sainor) was added dropwise at 0°C and stirred for 30 minutes. The reaction mixture was slowly heated to rt and stirred for 16 hours. Subsequently, the reaction mixture was quenched with 1N HCl at 0°C and extracted with ELISA. The combined organic extracts were washed with brine, separated, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified using a Redi-Sep pre-packed silica gel column (40 g) eluted with 0-10% MeOH in DCM to obtain tert-butyl(S)-6-(4-((1-hydroxy-5-methylhexane-3-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (1.4 g, 2.96 mmol, yield 72%) as a white solid. m / z(ESI):473.9(M+H) + .

[0244] Step 5: tert-butyl(S)-6-(4-((5-methyl-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate To a solution of tert-butyl(S)-6-(4-((1-hydroxy-5-methylhexane-3-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.90 g, 1.9 mmol) in DMSO (9 mL), TEA (2.65 mL, 19.0 mmol) and sulfur trioxide pyridine (0.91 g, 5.7 mmol, Chempure) were added. The reaction mixture was stirred in rt for 1 hour, then diluted with ice-cold water and extracted with ethyl acetate. The combined organic extracts were washed with brine, separated, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain tert-butyl(S)-6-(4-((5-methyl-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate as a colorless liquid, which was then transferred to the next step without purification. m / z(ESI):471.9(M+H) + .

[0245] Step 6: tert-butyl(S)-6-(4-((1-(1H-imidazole-2-yl)-4-methylpentan-2-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3,4]octane-2-carboxylate A solution of tert-butyl(S)-6-(4-((5-methyl-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazolin-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.30 g, 0.64 mmol), 7M ammonia (0.24 mL, 1.65 mmol), and oxalaldehyde (0.12 g, 0.83 mmol) in MeOH was stirred in a sealed tube of rt for 16 hours. The reaction mixture was then concentrated under reduced pressure. The residue was dissolved in water and extracted with ethyl acetate. The combined organic extracts were washed with brine, separated, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by reverse-phase column chromatography using 0-100% MeCN in water to obtain tert-butyl(S)-6-(4-((1-(1H-imidazole-2-yl)-4-methylpentan-2-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.07 g, 0.137 mmol, yield 22%) as a grayish-white solid. 1 H NMR(400MHz,DMSO-d6):δ ppm 11.78(s,1H),6.90(s,2H),6.32(s,1H),4.59(s,1H),3.77(s,4H),3.50-3.61(m,2H ),3.3(m,2H),2.76-2.91(m,2H),2.39(t,J=5.9Hz,2H),2.21(s,2H),2.06(t,J=6.9H z,2H),1.68(s,4H),1.59(q,J=6.6Hz,1H),1.49(dt,J=14.1,7.3Hz,1H),1.39(s,9H) ),1.13(dt,J=13.5,7.1Hz,1H),0.85(dd,J=13.3,6.5Hz,6H).m / z(ESI):510.9(M+H) + .

[0246] Additional step 3a of Example 1-24. [ka] Step 3a: tert-butyl7-(hydroxymethyl)-6-(4-(((S)-5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3,4]octane-2-carboxylate To a solution of 2-(tert-butyl)7-ethyl6-(4-(((S)-5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2,7-dicarboxylate (1.9 g, 3.32 mmol) in THF (20 mL) and MeOH (1 mL), LiBH4 (4.2 mL, 8.3 mmol, Symax) was added at -5°C. Subsequently, the reaction mixture was slowly heated to rt, stirred for 3 hours, and then diluted with a saturated aqueous solution of NH4Cl at -78°C and extracted with ELISA. The combined organic extract was washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified using a Redi-Sep pre-packed silica gel column (40 g) eluted with a gradient of 10-20% MeOH in DCM to obtain tert-butyl 7-(hydroxymethyl)-6-(4-(((S)-5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (1.3 g, 2.45 mmol, yield 74%) as a grayish-white solid. m / z(ESI): 530.8(M+H) +A diastereomer mixture (0.7g) is used in ChiralPak with 0.5% diethylamine (1:1) in liquid CO2:isopropyl alcohol. Separation by IC column yielded tert-butyl(S)-7-(hydroxymethyl)-6-(4-(((S)-5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.28 g, 0.53 mmol, yield 40%) as peak 1, and tert-butyl(R)-7-(hydroxymethyl)-6-(4-(((S)-5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.26 g, 0.49 mmol, yield 37%) as peak 2. The stereochemistry of the structure has been arbitrarily assigned and is not yet determined. Peak 1: 1 H NMR(400MHz,DMSO-d6):δ ppm 7.80(brs,1H),7.14-7.31(m,2H),6.19(br s,1H),4.53(m,1H),4.01(m,1H),3.80(m,5H),3.56(d,J=11.4Hz,2H),2.33(m,4H),2.06-2. 29(m,4H),1.64(m,6H),1.38(s,9H),1.22(m,1H),0.73-0.96(m,6H).m / z(ESI):530.8(M+H) + Peak 2: 1 H NMR(400MHz,DMSO-d6):δ ppm 7.83(br s,1H),7.14-7.29(m,2H),6.20(br s,1H),4.53(m,1H),3.48-3.90(m,7H),2.56(m,2H),2.27-2.45(m,4H),2.17(m,3H) ),1.61(m,6H),1.38(s,9H),1.17-1.29(m,1H),0.87(m,6H).m / z(ESI):530.8(M+H) + .

[0247] Additional step 3a of Example 1-26. [ka] Step 3a: tert-butyl(S)-6-(7-cyano-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)quinazoline-2-yl)-2,6-diazaspiro[3,4]octane-2-carboxylate To a solution of tert-butyl(S)-6-(7-bromo-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)quinazolin-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.40 g, 0.70 mmol), Zn(CN)2 (0.16 g, 1.39 mmol, Chempure), and Na2CO3 (0.22 g, 2.09 mmol) in DMF (8 mL), CuI (0.013 g, 0.07 mmol) and Pd(PPh3)4 (0.080 g, 0.07 mmol, Chempure) were added, and the reaction mixture was heated at 100 °C for 24 hours. The reaction mixture was diluted with water and filtered. The filtrate was extracted with ELISA. The organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified using a Redi-Sep pre-packed silica gel column (12 g) eluted with a gradient of 5-10% MeOH in DCM to obtain tert-butyl(S)-6-(7-cyano-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)quinazolin-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.35 g, 0.671 mmol, yield 97%) as a yellow solid. m / z(ESI): 521.8(M+H) + .

[0248] Additional steps 3a and 3b for Examples 1-59 and 1-36. [ka] Step 3a: (S)-(2-(2-(tert-butoxycarbonyl)-2,6-diazaspiro[3,4]octan-6-yl)-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)quinazoline-7-yl)boronic acid To a degassed solution of tert-butyl(S)-6-(7-bromo-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)quinazolin-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.50 g, 0.87 mmol), bis(pinacolato)diborone (0.33 g, 1.30 mmol, Chempure), and KOAc (0.256 g, 2.61 mmol) in 1,4-dioxane (5 mL), PdCl2(dppf)-DCM adduct (0.071 g, 0.087 mmol) was added, and the reaction mixture was stirred at 100 °C for 16 hours. The reaction mixture was diluted with water and filtered through a Celite pad. The filtrate was extracted with ELISA. The organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain (S)-(2-(2-(tert-butoxycarbonyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)quinazoline-7-yl)boronic acid, which was used in the next step without purification. m / z(ESI): 540.8(M+H) + .

[0249] Step 3b: tert-butyl(S)-6-(4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)-7-(thiazole-2-yl)quinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate PdCl2(dppf)-DCM adduct (0.091g, 0.11 mmol) was added to a degassed solution of (S)-(2-(2-(tert-butoxycarbonyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)quinazoline-7-yl)boronic acid (0.30 g, 0.56 mmol), 2-bromothiazole (0.11 g, 0.67 mmol, Combi-Blocks), and K2CO3 (0.31 g, 2.22 mmol) in 1,4-dioxane (1.5 mL) and water (0.6 mL). The reaction mixture was heated at 100 °C for 16 hours. The reaction mixture was diluted with water and filtered through a Celite pad. The filtrate was extracted with ELISA. The organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by reverse-phase column chromatography to obtain tert-butyl(S)-6-(4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)-7-(thiazole-2-yl)quinazolin-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate as a yellow solid. m / z(ESI): 579.8(M+H) + .

[0250] Additional step 3a of Example 1-38. [ka] Step 3a: tert-butyl(S)-6-(7-cyclopropyl-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)pyrido[3,2-d]pyrimidine-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate 1,4-Dioxane (1.6 mL) and water (0.08 mL) containing tert-butyl(S)-6-(7-bromo-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)pyrido[3,2-d]pyrimidine-2-yl)-2,6-diazaspiro[3,4]octane-2-carboxylate (0.20 g, 0.35 mmol), cyclopropyltrifluoro-λ 4To a degassed solution of borane, potassium salt (0.103 g, 0.695 mmol, Combi-Blocks), and Na2CO3 (0.11 g, 1.04 mmol), PdCl2(dppf)-DCM adduct (0.028 g, 0.035 mmol, Chempure) was added. The reaction mixture was heated to 90°C for 16 hours, then diluted with water and extracted with DCM. The organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified using a Redi-Sep pre-packed silica gel column (10 g) eluted with a gradient of 5-10% MeOH in DCM, yielding tert-butyl(S)-6-(7-cyclopropyl-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)pyrido[3,2-d]pyrimidine-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.105 g, 0.195 mmol, yield 56%) as a brown solid. m / z(ESI): 537.9(M+H) + .

[0251] Additional step 3a of Examples 1-40. [ka] Step 3a: tert-butyl(S)-6-(7-cyclopropyl-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)quinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate 1,4-Dioxane (1.6 mL) and water (0.08 mL) containing tert-butyl(S)-6-(7-bromo-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)quinazolin-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.20 g, 0.35 mmol), cyclopropyltrifluoro-λ 4- A degassed solution of borane, potassium salt (0.103 g, 0.695 mmol, Combi-Blocks) and Na2CO3 (0.11 g, 1.02 mmol) was mixed with PdCl2(dppf)-DCM adduct (0.028 g, 0.035 mmol, Chempure). The reaction mixture was heated at 90°C for 16 hours, then diluted with water and extracted with DCM. The organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified using a Redi-Sep pre-packed silica gel column (4g) eluted with a gradient of 5-10% MeOH in DCM to obtain tert-butyl(S)-6-(7-cyclopropyl-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)quinazolin-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.11g, 0.205 mmol, yield 59%) as a brown solid. m / z(ESI): 536.8(M+H) + .

[0252] Additional steps 3a to 3c of Example 1-42. [ka] Step 3a: (S)-(2-(2-(tert-butoxycarbonyl)-2,6-diazaspiro[3,4]octan-6-yl)-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)pyrido[3,2-d]pyrimidine-7-yl)boronic acid To a degassed solution of tert-butyl(S)-6-(7-bromo-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)pyrido[3,2-d]pyrimidine-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (250 mg, 0.434 mmol), bis(pinacolato)diborone (165 mg, 0.650 mmol, Chempure), and KOAc (128 mg, 1.30 mmol) in 1,2-dimethoxyethane (5 mL), PdCl2(dppf)-DCM adduct (26.5 mg, 0.033 mmol) was added, and the reaction mixture was stirred at 80°C for 6 hours. The reaction mixture was diluted with water and filtered through a Celite pad. The filtrate was extracted with ELISA. The organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain (S)-(2-(2-(tert-butoxycarbonyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)pyrido[3,2-d]pyrimidine-7-yl)boronic acid (230 mg, 0.43 mmol, yield 98%), which was then transferred to the next step without purification. m / z(ESI): 541.8(M+H) + .

[0253] Step 3b: tert-butyl(S)-6-(7-hydroxy-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)pyrido[3,2-d]pyrimidine-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate To a suspension of (S)-(2-(2-(tert-butoxycarbonyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)pyrido[3,2-d]pyrimidine-7-yl)boronic acid (130 mg, 0.24 mmol) and sodium hydroxide (1.5 mL, 3.00 mmol) in THF (5 mL), hydrogen peroxide (0.8 mL, 7.8 mmol) was added at 0°C, and the resulting mixture was stirred at rt for 2 hours. Next, the reaction mixture was diluted with water and extracted with 5% MeOH in DCM. The combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified using a Redi-Sep pre-packed silica gel column (4g) eluted with a gradient of 0-10% MeOH and DCM to obtain tert-butyl(S)-6-(7-hydroxy-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)pyrido[3,2-d]pyrimidine-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.12g, 0.22 mmol). m / z(ESI): 513.9(M+H) + .

[0254] Step 3c: tert-butyl(S)-6-(7-methoxy-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)pyrido[3,2-d]pyrimidine-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate To a suspension of tert-butyl(S)-6-(7-hydroxy-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)pyrido[3,2-d]pyrimidine-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.12 g, 0.22 mmol) and Na2CO3 (0.12 g, 1.12 mmol) in DMF (3 mL), iodomethane (0.07 mL, 1.12 mmol) was added, and the resulting mixture was stirred at rt for 16 hours. Subsequently, the reaction mixture was diluted with water and extracted with SiO2. The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified using a Redi-Sep pre-packed silica gel column (4 g) eluted with a gradient of 0-10% MeOH and DCM to obtain tert-butyl(S)-6-(7-methoxy-4-((5-methyl-1-(methylamino)-1-oxohexane-3-yl)amino)pyrido[3,2-d]pyrimidine-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.080 g, 0.152 mmol). m / z(ESI): 527.8(M+H) + .

[0255] Additional steps 3a and 3b of Example 1-57. [ka] Step 3a: tert-butyl(S)-6-(4-((1-((2,2-dimethoxyethyl)amino)-5-methyl-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3,4]octane-2-carboxylate To a solution of (S)-3-((2-(2-(tert-butoxycarbonyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydroquinazoline-4-yl)amino)-5-methylhexanoic acid (0.50 g, 1.03 mmol, step 3 of Examples 1-22), EDCI (0.24 g, 1.23 mmol, Chempure), and HOBt (0.19 g, 1.23 mmol, Chempure), 2,2-dimethoxyethane-1-amine (0.16 g, 1.54 mmol, Avra) was added, and the resulting mixture was heated at 60°C for 16 hours. The reaction mixture was diluted with ice-cold water and extracted with ethyl acetate. The combined organic extracts were washed with brine, separated, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain tert-butyl(S)-6-(4-((1-((2,2-dimethoxyethyl)amino)-5-methyl-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.40 g), which was used in the next step without purification. m / z(ESI): 575.0(M+H) + .

[0256] Step 3b: (S)-N-(4-methyl-1-(oxazol-2-yl)pentan-2-yl)-2-(2,6-diazaspiro[3,4]octan-6-yl)-5,6,7,8-tetrahydroquinazoline-4-amine A solution of tert-butyl(S)-6-(4-((1-((2,2-dimethoxyethyl)amino)-5-methyl-1-oxohexane-3-yl)amino)-5,6,7,8-tetrahydroquinazolin-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.40 g, 0.70 mmol) in phosphorus pentoxide (Eaton's reagent) (4.0 mL, 0.696 mmol, Alfa-Aesar) in methanesulfonic acid was heated at 100°C for 16 hours. The reaction mixture was diluted with ice-cold water and extracted with ethyl acetate. The combined organic extracts were washed with brine, separated, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by reverse-phase column chromatography using a C18 column with a gradient of 0-15% MeCN in H2O to obtain (S)-N-(4-methyl-1-(oxazol-2-yl)pentan-2-yl)-2-(2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydroquinazoline-4-amine (0.12 g, 0.292 mmol, yield 42%) as a brown solid. m / z(ESI):411.0(M+H) + .

[0257] Alternative step 4 of Examples 1-58. [ka] Step 4: (3S)-N,5-dimethyl-3-((7-methyl-2-(2-(2-(trifluoromethyl)-2-propenoyl)-2,6-diazaspiro[3,4]octan-6-yl)-4-quinazolinyl)amino)hexaneamide To a solution of (S)-N,5-dimethyl-3-((7-methyl-2-(2,6-diazaspiro[3.4]octan-6-yl)quinazolin-4-yl)amino)hexaneamide hydrochloride (0.15 g, 0.34 mmol), TEA (0.37 mL, 2.68 mmol, Combi-Blocks) in DCM, T3P (0.641 g, 1.01 mmol, 50% solution in ethyl acetate, Spectrochem) was added at 0°C. The resulting reaction mixture was stirred in rt for 3 hours, then diluted with ice-cold water and extracted with ethyl acetate. The combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC using an XBridge Prep C18 5μm OBD column (150×30mm, 0.1% NH4HCO3 in MeCN / H2O, gradient 35%~80% over 15 minutes, flow rate = 30mL / min) to obtain (S)-N,5-dimethyl-3-((7-methyl-2-(2-(2-(trifluoromethyl)acryloyl)-2,6-diazaspiro[3.4]octan-6-yl)quinazoline-4-yl)amino)hexaneamide (0.008g, 0.015 mmol, yield 4.5%) as a white solid. 1 H NMR(400MHz,DMSO-d6):δ ppm 7.89(s,1H),7.79(s,1H),7.12(s,1H),6.92(s,1H),6.34-6.47(m,1H),6.26(s,1H),4.77(br s,1H),4.16-4.31(m,2H),3.98(brs,2H),3.49-3.86(m,4H),2.54(s,3H),2.40-2.49(m,1H),2.28-2.39(m,4H),2.18(br s,2H),1.54-1.72(m,2H),1.26-1.38(m,1H),0.82-0.97(m,6H). 19 F NMR(376MHz,DMSO- d6)δ ppm -63.79,-73.41.m / z(ESI):533.2(M+H) + .

[0258] Additional step 3a of Examples 1-60. [ka] Step 3a: tert-butyl(S)-6-(4-((4-methyl-1-(methylamino)-1-oxopentan-2-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3,4]octane-2-carboxylate A mixture of tert-butyl(S)-6-(4-((1-methoxy-4-methyl-1-oxopentan-2-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.30 g, 0.62 mmol) and a solution of 1N methaneamine in MeOH (3.0 mL, 6.15 mmol, Spectrochem) was stirred in a sealed tube of rt for 16 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified using a Redi-Sep pre-packed silica gel column (12 g) eluted with 30-50% ethyl hexane to obtain tert-butyl(S)-6-(4-((4-methyl-1-(methylamino)-1-oxopentan-2-yl)amino)-5,6,7,8-tetrahydroquinazoline-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.25 g, 0.514 mmol, yield 84%) as a grayish-white solid. m / z(ESI):486.9(M+H) + .

[0259] Synthesis of intermediates Intermediate 1: 2,4-dichloro-8-methyl-5,6,7,8-tetrahydroquinazoline [ka] Step 1: Ethyl 3-methyl-2-oxocyclohexane-1-carboxylate A mixture of ethyl 2-oxocyclohexane-1-carboxylate (20.0 g, 117.6 mmol, ADAMAS) in THF (300 mL) was stirred at 0°C, and a solution of LDA (117.6 mL, 2.0 M) in THF was added dropwise. The mixture was stirred for 0.5 hours. Methyl iodide (16.7 g, 117.6 mmol) was added, and the resulting mixture was stirred for 1 hour. A saturated solution of NH4Cl was added, and the mixture was extracted with SiO2 (200 mL x 3). The combined organic phase was washed with brine, dried over sodium sulfate, and concentrated to obtain crude ethyl 3-methyl-2-oxocyclohexane-1-carboxylate (21.0 g, 114 mmol, yield 97%) as a yellow oil, which was used in the next step without further purification.

[0260] Step 2: 8-methyl-5,6,7,8-tetrahydroquinazoline-2,4(1H,3H)-dione In a 250 mL round-bottom flask, ethyl 3-methyl-2-oxocyclohexane-1-carboxylate (21.0 g, 114 mmol), urea (10.3 g, 171 mmol), sodium methoxide (6.3 g, 171 mmol), and EtOH (150 mL) were added. The reaction mixture was stirred at 80 °C for 16 hours. The solution was concentrated under vacuum to obtain the crude product. DCM (100 mL) was added, the mixture was filtered, the solid was washed with DCM (50 mL x 3), and dried to obtain 8-methyl-5,6,7,8-tetrahydroquinazoline-2,4(1H,3H)-dione (10.0 g, 55.5 mmol, yield 48%) as a brown solid. m / z(ESI,+ve ion):181.0(M+H)+.

[0261] Step 3: 2,4-Dichloro-8-methyl-5,6,7,8-tetrahydroquinazoline A mixture of 8-methyl-5,6,7,8-tetrahydroquinazoline-2,4(1H,3H)-dione (7.8 g, 43.3 mmol) and POCl3 (80 mL) was heated to 100°C and stirred for 16 hours. The mixture was cooled to room temperature, and the residue was adjusted to pH 8-9 with a saturated solution of NaHCO3 at 0°C. The mixture was extracted with DCM (100 mL x 3), the combined organic phase was washed with brine, dried over sodium sulfate, and concentrated. The residue was purified by silica gel chromatography (PE:siRNA = 20:1) to obtain 2,4-dichloro-8-methyl-5,6,7,8-tetrahydroquinazoline (3.0 g, 13.8 mmol, yield 32%) as a yellow solid. m / z (ESI, +ve ion): 217.1 (M + H) + . 1H NMR:(400Hz,CDCl3):δ ppm,2.98-2.93(m,1H),2.73(t,J=6.0Hz,2H),2.03-1.91(m,2H),1.82-1.77(m,1H),1.66-1.59(m,1H),1.37(d,J=7.2Hz,3H).

[0262] [Table 22]

[0263] Alternative step 1 (for intermediates 2-5): Methyl 4,4-dimethyl-2-oxocyclohexane-1-carboxylate [ka] Step 1: Methyl 4,4-dimethyl-2-oxocyclohexane-1-carboxylate In a solution of 3,3-dimethylcyclohexane-1-one (25.0 g, 198 mmol, Combi-Blocks) and dimethyl carbonate (44.6 g, 495 mmol) in 0°C THF (250 mL), NaH (19.8 g, 495 mmol) was added in small increments, and the mixture was heated to rt and then heated under reflux for 16 hours. The reaction mass was poured into an ice-cold saturated NH4Cl solution and extracted with ELISA. The combined organic extracts were washed with brine, separated, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain methyl-4,4-dimethyl-2-oxocyclohexane-1-carboxylate (36.5 g, 198 mmol, 100% yield) as a pale yellow liquid. m / z(ESI): 185.1(M+H) + .

[0264] Intermediate 6: 2,4-dichloro-7,8-dihydro-6H-pyrano[3,2-d]pyrimidine [ka] Step 1: Ethyl 2-diazo-6-hydroxy-3-oxohexanoate A solution of 2-diazoethyl acetate (6.75 mL, 63.9 mmol, TCI) and dihydrofuran-2(3H)-one (4.42 mL, 58.1 mmol, Chempure) in THF (75 mL) at -78°C was to be to which a freshly prepared solution of LDA in THF (65.8 mL, 99 mmol, 1.5 M in THF) was added dropwise. The solution was stirred at -78°C for 30 minutes, and then acetic acid (21.9 mL, 383 mmol) was added dropwise to the reaction mixture at -78°C. The reaction mixture was diluted with ice-cold water and extracted with SiO2. The organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified using a Redi-Sep pre-packed silica gel column (40 g) eluting with a gradient of 20-30% ethyl in PE, yielding ethyl 2-diazo-6-hydroxy-3-oxohexanoate (2.0 g, 9.99 mmol, yield 17%) as a brown liquid. 1¹H NMR (400 MHz, chloroform-d): δ ppm 4.14-4.53 (m, 2H), 3.70 (t, J=6.1 Hz, 2H), 3.00 (t, J=7.0 Hz, 2H), 1.94 (m, 2H), 1.27-1.45 (m, 3H).

[0265] Step 2: Ethyl-3-oxotetrahydro-2H-pyran-2-carboxylate A suspension of rhodium(II) acetate dimer (0.088 g, 0.20 mmol) in toluene (30 mL) was to be mixed with a solution of ethyl 2-diazo-6-hydroxy-3-oxohexanoate (2.0 g, 9.99 mmol) in toluene (30 mL) over 90 minutes at 90°C. The mixture was then stirred at 90°C for 30 minutes, followed by concentration under reduced pressure. The residue was purified using a Redi-Sep pre-packed silica gel column (40 g) eluted with a gradient of 10% ethyl ethyl 3-oxotetrahydro-2H-pyran-2-carboxylate (700 mg, 4.1 mmol, yield 40%) as a pale yellow liquid. 1 ¹H NMR (400 MHz, chloroform-d): δ ppm 4.18-4.48 (m, 2H), 3.78-4.04 (m, 2H), 2.40 (t, J=6.7 Hz, 2H), 1.87-2.01 (m, 2H), 1.13-1.51 (m, 3H).

[0266] Step 3: 7,8-dihydro-6H-pyrano[3,2-d]pyrimidine-2,4-diol To a solution of ethyl 3-oxotetrahydro-2H-pyran-2-carboxylate (5.0 g, 29.0 mmol) in EtOH (50 mL), urea (1.74 g, 29.0 mmol, Avra) and sodium ethoxide (14.5 mL, 43.6 mmol, 21% solution in EtOH, Symacs) were added, and the reaction mixture was heated at 80°C for 16 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by reverse-phase column chromatography using a C18 column eluting with a gradient of 0-10% MeCN in H2O to obtain 7,8-dihydro-1H-pyrano[3,2-d]pyrimidine-2,4(3H,6H)-dione (4.7 g, 28.0 mmol, yield 96%) as a light brown solid.1 H NMR(400MHz,D2O):δ ppm 3.81-3.50(m,2H),2.20-2.24(m,2H),1.65-1.71(m,2H).m / z(ESI):169.1(M+H) + .

[0267] Step 4: 2,4-Dichloro-7,8-dihydro-6H-pyrano[3,2-d]pyrimidine A solution of 7,8-dihydro-1H-pyrano[3,2-d]pyrimidine-2,4(3H,6H)-dione (500 mg, 2.97 mmol) and N,N-dimethylaniline (360 mg, 2.97 mmol) in POCl3 (10 mL, 107 mmol) was heated at 80°C for 16 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified using a Redi-Sep pre-packed silica gel column (12 g) eluting with a 25-30% siRNA gradient in PE to obtain 2,4-dichloro-7,8-dihydro-6H-pyrano[3,2-d]pyrimidine (150 mg, 0.73 mmol, yield 24%) as a pale yellow liquid. m / z(ESI): 206.9(M+H) + .

[0268] Intermediate 7: 2,4-dichloro-7-methylpyrido[3,2-d]pyrimidine [ka] Step 1: 7-Methylpyrido[3,2-d]pyrimidine-2,4-diol To a degassed solution of 3-amino-5-methylpicolinonitrile (10.0 g, 75.0 mmol, Arbor) in DMF (100 mL), DBU (11.3 mL, 75 mmol, Spectrochem) was added at rt. The reaction mixture was stirred at 105 °C for 5 hours and then at rt for 12 hours under a CO2 atmosphere. Subsequently, the reaction mixture was cooled to 0 °C and acidified with 1.5 N HCl. The precipitated solid was filtered, washed with ethyl acetate, and dried under reduced pressure to obtain 7-methylpyrido[3,2-d]pyrimidine-2,4-diol (7.6 g, 42.9 mmol, yield 57%) as a grayish-white solid, which was then carried to the next step without purification. 1H NMR(400MHz,DMSO-d6):δ ppm 11.40(s,1H),11.17(s,1H),8.30(s,1H),7.33(s,1H),2.37(s,3H).m / z(ESI):178.0(M+H) + .

[0269] Step 2: 2,4-Dichloro-7-methylpyrido[3,2-d]pyrimidine To a solution of 7-methylpyrido[3,2-d]pyrimidine-2,4-diol (4.0 g, 22.6 mmol) in POCl3 (40 mL, 429 mmol), PCl5 (18.8 g, 90.0 mmol) was added, and the reaction mixture was stirred at 135 °C for 15 hours. The reaction mixture was concentrated under vacuum, and the residue was diluted with DCM. Subsequently, ice-cold water was added to the resulting solution, and extraction was performed with DCM. The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified using a Redi-Sep pre-packed silica gel column (120 g) eluted with a 30-50% siRNA gradient in hexane to obtain 2,4-dichloro-7-methylpyrido[3,2-d]pyrimidine (1.7 g, 7.94 mmol, yield 35%) as a grayish-white solid. 1 H NMR(400MHz,DMSO-d6):δ ppm 9.07(d,J=1.6Hz,1H),8.27(d,J=1.6Hz,1H),2.61(s,3H).m / z(ESI):215.9(M+H) + .

[0270] Synthesis of amines: Amine 1:(S)-4-methyl-1-(4H-1,2,4-triazole-3-yl)pentan-2-amine hydrochloride [ka] Step 1: tert-butyl(S)-(1-amino-5-methyl-1-oxohexane-3-yl)carbamate To a solution of (S)-3-((tert-butoxycarbonyl)amino)-5-methylhexanoic acid (5.0 g, 20.4 mmol, Angene) in 1,4-dioxane (40 mL), (Boc)2O (6.15 mL, 26.5 mmol, Spectrochem), pyridine (0.99 mL, 12.2 mmol), and ammonium bicarbonate (1.93 g, 24.5 mmol, Chempure) were added. The reaction mixture was stirred in rt for 16 hours, then diluted with water and extracted with ethyl acetate. The combined organic extract was washed with brine, separated, dried over Na2SO4, filtered, and concentrated under vacuum to obtain tert-butyl(S)-(1-amino-5-methyl-1-oxohexane-3-yl)carbamate (4.8 g, 19.7 mmol, 96% yield), which was used in the next step without purification. m / z(ESI):145.2(M+H-Boc) + .

[0271] Step 2: tert-butyl(S)-(4-methyl-1-(4H-1,2,4-triazole-3-yl)pentan-2-yl)carbamate A solution of tert-butyl(S)-(1-amino-5-methyl-1-oxohexane-3-yl)carbamate (2.0 g, 8.2 mmol) in 1,1-dimethoxy-N,N-dimethylmethaneamine (10 mL, 8.19 mmol, Spectrochem) was heated at 100°C for 1 hour. The reaction mixture was then concentrated under reduced pressure. The residue was redissolved in AcOH (20 mL) and hydrazine hydrate (1.0 mL, 31.9 mmol, Spectrochem) was added. The reaction mixture was heated at 100°C for 1 hour. The reaction mixture was then concentrated under reduced pressure, the residue was diluted with water, and extracted with DCM. The organic extract was washed with saturated NaHCO3 and brine, separated, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain tert-butyl(S)-(4-methyl-1-(1H-1,2,4-triazole-5-yl)pentan-2-yl)carbamate (1.8 g, 6.71 mmol, yield 82%), which was then transferred to the next step without purification. m / z(ESI):269.1(M+H) + .

[0272] Step 3: (S)-4-methyl-1-(4H-1,2,4-triazole-3-yl)pentan-2-amine hydrochloride A solution of tert-butyl(S)-(4-methyl-1-(1H-1,2,4-triazole-5-yl)pentan-2-yl)carbamate (2.3 g, 8.6 mmol) in 1,4-dioxane (10 mL) was treated with 4 M HCl (21.4 mL, 86.0 mmol, Symax) in dioxane. The resulting solution was stirred at rt for 16 hours, then concentrated under reduced pressure and triturated with Et2O to obtain (S)-4-methyl-1-(1H-1,2,4-triazole-5-yl)pentan-2-amine hydrochloride, which was used without further purification. m / z(ESI): 169.2(M+H) + .

[0273] Amine 2: Methyl 3-amino-3-(3-cyanophenyl)propanoate [ka] Step 1: 3-amino-3-(3-cyanophenyl)propanoic acid To a solution of 3-formylbenzonitrile (8.0 g, 61.0 mmol, Combi-Blocks) in EtOH (100 mL), malonic acid (6.35 g, 61.0 mmol) and ammonium acetate (9.41 g, 122 mmol) were added. The resulting mixture was stirred at 90°C for 16 hours, then filtered and dried to obtain 3-amino-3-(3-cyanophenyl)propanoic acid (7.1 g, 37.3 mmol, yield 61%) as a white solid. m / z(ESI): 191.1(M+H) + .

[0274] Step 2: Methyl 3-amino-3-(3-cyanophenyl)propanoate To a solution of 3-amino-3-(3-cyanophenyl)propanoic acid (7.1 g, 37.3 mmol) in MeOH (100 mL), SOCl2 (5.45 mL, 74.7 mmol) was added, and the reaction mixture was stirred at 60°C for 16 hours. Subsequently, the reaction mixture was concentrated and co-distilled with toluene to obtain methyl 3-amino-3-(3-cyanophenyl)propanoate (7.6 g, 37.2 mmol, 100% yield) as a pale yellow solid. m / z(ESI): 205.2(M+H) + .

[0275] Amine 3:methyl(S)-3-amino-5-methylhexanoate hydrochloride [ka] Step 1: Methyl(S)-3-((tert-butoxycarbonyl)amino)-5-methylhexanoate To a solution of (S)-3-((tert-butoxycarbonyl)amino)-5-methylhexanoic acid (20.0 g, 82.0 mmol, Enamine) in DMF (100 mL), cesium carbonate (34.5 g, 106 mmol) and methyl iodide (7.65 mL, 122 mmol, Spectrochem) were added. The reaction mixture was stirred at 50°C for 16 hours, then diluted with ice-cold water and extracted with ethyl acetate. The organic extract was washed with brine, separated, dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified using a Redi-Sep pre-packed silica gel column (40 g) eluting with a 40-45% ethyl acetate gradient in hexane to obtain methyl(S)-3-((tert-butoxycarbonyl)amino)-5-methylhexanoate (21.0 g, 81.0 mmol, 99% yield) as a pale yellow oil. 1 H NMR (400MHz, chloroform-d): δ ppm 4.88(d,J=9.4Hz,1H),4.01(s,1H),3.70(s,3H),2.53(m,2H),1.63-1.72(m,1H), 1.45(s,9H),1.29(m,2H),0.94(dd,J=6.6,4.3Hz,6H).m / z(ESI):160.2(M+H-Boc) + .

[0276] Step 2: Methyl(S)-3-amino-5-methylhexanoate hydrochloride To a solution of methyl(S)-3-((tert-butoxycarbonyl)amino)-5-methylhexanoate (21.0 g, 81.0 mmol) in 1,4-dioxane (100 mL), 4 M HCl (22.3 mL, 89.0 mmol, Spectrochem) was added to the dioxane and the mixture was stirred at rt for 30 minutes. Subsequently, the reaction mixture was concentrated under vacuum to obtain methyl(S)-3-amino-5-methylhexanoate hydrochloride (15.5 g, 79 mmol, yield 98%) as a viscous liquid. 1 H NMR (400MHz, chloroform-d): δ ppm 4.88(d,J=9.4Hz,1H),4.01(s,1H),3.70(s,3H),2.53(m,2H),1.63-1.72(m,1H ),1.45(s,9H),1.29(m,2H),0.94(dd,J=6.6,4.3Hz,6H).m / z(ESI):160.2(M+H) + .

[0277] Amine 4:(S)-4-methyl-1-(3-methyl-1,2,4-oxadiazole-5-yl)pentan-2-amine hydrochloride [ka] Step 1: tert-butyl(S)-(1-(acetimidoamideoxy)-5-methyl-1-oxohexane-3-yl)carbamate A solution of (S)-3-((tert-butoxycarbonyl)amino)-5-methylhexanoic acid (2.0 g, 8.15 mmol), N-hydroxyacetimamide (0.66 g, 8.97 mmol, TCI), EDC-HCl (1.88 g, 9.78 mmol, Chempure), and HOBt (1.37 g, 8.97 mmol, Avra) in 20 mL of DCM was stirred at rt for 16 hours. The reaction mixture was then diluted with water and extracted with DCM. The combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain tert-butyl(S)-(1-(acetimidoamideoxy)-5-methyl-1-oxohexanoyl)carbamate. m / z(ESI): 302.1(M+H) + .

[0278] A solution of tert-butyl(S)-(1-(acetimidoamideoxy)-5-methyl-1-oxohexane-3-yl)carbamate (2.0 g, 6.64 mmol) and KOAc (0.716 g, 7.30 mmol) in DMF (20 mL) was heated in a microwave reactor at 120 °C (Personal Chemistry, Biotage AB, Inc., Upsala, Sweden) for 120 minutes. The reaction mixture was then diluted with water and extracted with SiO2. The combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified using a Redi-Sep pre-packed silica gel column (40 g) eluting with a gradient of 0-15% ethyl acetate in PE to obtain tert-butyl(S)-(4-methyl-1-(3-methyl-1,2,4-oxadiazole-5-yl)pentan-2-yl)carbamate (1.1 g, 3.88 mmol, yield 58%). m / z(ESI): 284.1(M+H) + .

[0279] Step 2: (S)-4-methyl-1-(3-methyl-1,2,4-oxadiazole-5-yl)pentan-2-amine hydrochloride A solution of tert-butyl(S)-(4-methyl-1-(3-methyl-1,2,4-oxadiazole-5-yl)pentan-2-yl)carbamate (0.6 g, 2.12 mmol) in 1,4-dioxane (10 mL) was treated with 4 M HCl (6.0 mL, 2.12 mmol, Symax) in dioxane. The resulting solution was stirred at rt for 16 hours, then concentrated under reduced pressure and triturated with Et2O to obtain (S)-4-methyl-1-(3-methyl-1,2,4-oxadiazole-5-yl)pentan-2-amine hydrochloride, which was used without further purification. m / z(ESI): 184.1(M+H) + .

[0280] Amine 5:tert-butyl 8-fluoro-2,6-diazaspiro[3,4]octane-2-carboxylate [ka] Step 1: tert-butyl6-benzyl-7-oxo-2,6-diazaspiro[3,4]octane-2-carboxylate A solution of tert-butyl 7-oxo-2,6-diazaspiro[3.4]octane-2-carboxylate (500 mg, 2.21 mmol) and benzyl bromide (397 mg, 2.32 mmol) was added to 10 mL of THF at 0°C, to which NaH (60% dispersion, 97 mg, 2.43 mmol) was added. The reaction mixture was stirred at 60°C for 18 hours. After completion, the reaction product was diluted with water and extracted with siRNA. The combined organic extract was washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was stirred in 15 mL of Et2O and filtered to obtain tert-butyl 6-benzyl-7-oxo-2,6-diazaspiro[3.4]octane-2-carboxylate (500 mg, 1.58 mmol, yield 72%) as a white solid. m / z(ESI):317.0(M+H) + .

[0281] Step 2: tert-butyl6-benzyl-8-fluoro-7-oxo-2,6-diazaspiro[3,4]octane-2-carboxylate A stirred solution of tert-butyl 6-benzyl-7-oxo-2,6-diazaspiro[3.4]octane-2-carboxylate (500 mg, 1.58 mmol) in THF (15 mL) was cooled to -15°C, and LHMDS (1 M in THF) (2.05 mL, 2.05 mmol) was added dropwise. The reaction mixture was stirred at -15°C for 45 minutes. Then, a solution of NFSI (997 mg, 3.16 mmol) in THF (5 mL) was added dropwise, and the mixture was stirred at the same temperature for 45 minutes. The reaction mixture was quenched with saturated ammonium chloride solution, extracted with ₹ (2 × 50 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was purified by column chromatography on 230-400 mesh silica gel using 40% toluene in PE to obtain tert-butyl6-benzyl-8-fluoro-7-oxo-2,6-diazaspiro[3.4]octane-2-carboxylate (330 mg, 0.987 mmol, yield 62%) as a colorless solid. m / z(ESI):235.1(M-Boc+H) + .

[0282] Step 3: tert-butyl 6-benzyl-8-fluoro-2,6-diazaspiro[3,4]octane-2-carboxylate To a stirred solution of tert-butyl6-benzyl-8-fluoro-7-oxo-2,6-diazaspiro[3.4]octane-2-carboxylate (100 mg, 0.30 mmol) in THF (6 mL), borane-THF complex (1 M, 1.2 mL, 1.20 mmol) was added dropwise under an N2 atmosphere at 0°C. The reaction mixture was then heated at 65°C for 4 hours. The reaction product was quenched with saturated ammonium chloride solution and extracted with siRNA (2 × 20 mL). The combined organic extract was dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was purified by column chromatography on 230-400 mesh silica gel using 28% siRNA in PE to obtain tert-butyl6-benzyl-8-fluoro-2,6-diazaspiro[3.4]octane-2-carboxylate (25 mg, 0.078 mmol, yield 26%) as a colorless solid. m / z(ESI):321.3(M+H) + .

[0283] Step 4: tert-butyl 8-fluoro-2,6-diazaspiro[3,4]octane-2-carboxylate To a solution of tert-butyl 6-benzyl-8-fluoro-2,6-diazaspiro[3.4]octane-2-carboxylate (400 mg, 1.25 mmol) in MeOH (10 mL), Pd-C (133 mg, 1.25 mmol) was added. The system was evacuated and H2 was refilled. The reaction mixture was stirred at rt for 2 hours. The reaction mixture was filtered and concentrated under vacuum to obtain tert-butyl 8-fluoro-2,6-diazaspiro[3.4]octane-2-carboxylate (270 mg, 1.17 mmol, yield 94%) as a grayish-white solid. m / z(ESI): 231.0(M+H) + .

[0284] Amine 6: (S)-4-methyl-1-(thiazole-2-yl)pentan-2-amine [ka] Step 1: tert-butyl(S)-(1-amino-5-methyl-1-thioxohexane-3-yl)carbamate To a solution of tert-butyl(S)-(1-amino-5-methyl-1-oxohexane-3-yl)carbamate (4.0 g, 16.4 mmol) in 1,4-dioxane (50 mL), Lawson's reagent (3.64 g, 9.0 mmol, Spectrochem) was added. The reaction mixture was stirred at 60°C for 2 hours, followed by 16 hours at rt. Next, the reaction mixture was diluted with water and extracted with DCM. The combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain tert-butyl(S)-(1-amino-5-methyl-1-oxohexane-3-yl)carbamate (3.8 g, 15.6 mmol, 95% yield) as a grayish-white solid. m / z(ESI): 261.0(M+H) + .

[0285] Step 2: Benzyl(S)-(1-amino-5-methyl-1-thioxohexane-3-yl)carbamate To a solution of tert-butyl(S)-(1-amino-5-methyl-1-thioxohexane-3-yl)carbamate (4.0 g, 15.36 mmol) in 1,4-dioxane (10 mL), HCl (38.4 mL, 154 mmol, 4 M solution, Symacs) was added to the dioxane, and the reaction mixture was stirred under rt for 16 hours. Subsequently, the reaction mixture was concentrated under reduced pressure to obtain (S)-3-amino-5-methylhexanethioamide hydrochloride (3.0 g, 15.3 mmol, 99% yield). m / z(ESI): 161.1(M+H) + .

[0286] To a solution of (S)-3-amino-5-methylhexanethioamide hydrochloride (3.0 g, 15.3 mmol) and NaHCO3 (2.82 g, 33.5 mmol) in 1,4-dioxane (21 mL) and water (45 mL), benzyl chloroformate (5.22 mL, 18.30 mmol, Chempure) was added in toluene at 0°C, and the resulting mixture was stirred at rt for 16 hours. Next, the reaction mixture was diluted with water and extracted with DCM. The combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified using a Redi-Sep pre-packed silica gel column (40 g) eluting with a gradient of 70-80% siRNA in hexane to obtain benzyl(S)-(1-amino-5-methyl-1-thioxohexane-3-yl)carbamate (3.0 g, 10.2 mmol, yield 67%) as a pale yellow oil. m / z(ESI):295.0(M+H) + .

[0287] Step 3: Benzyl(S)-(4-methyl-1-(thiazole-2-yl)pentan-2-yl)carbamate Bromoacetaldehyde diethyl acetal (3.85 mL, 25.4 mmol, Chempure) was added to concentrated HCl (5 mL, 165 mmol) and heated at 60°C for 30 minutes. This mixture was then cooled to 10°C. DMF (10 mL), followed by powdered molecular sieve (1 spatula), was added. The solution was decanted and immediately used as described below. The solution of bromoacetaldehyde in DMF prepared as described above was added to benzyl(S)-(1-amino-5-methyl-1-thioxohexane-3-yl)carbamate (3.0 g, 10.2 mmol) and heated at 60°C for 5 hours. Subsequently, the reaction mixture was cooled to rt, diluted with ethyl acetate, and washed with a saturated aqueous solution of sodium bicarbonate. The combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified using a Redi-Sep pre-packed silica gel column (40 g) eluted with a 30-40% siRNA gradient in hexane to obtain benzyl(S)-(4-methyl-1-(thiazole-2-yl)pentan-2-yl)carbamate (1.5 g, 4.71 mmol, yield 46%) as a pale yellow oil. 1 H NMR(400MHz,DMSO-d6)δ ppm 7.70(d,J=3.6Hz,1H),7.57(d,J=3.6Hz,1H),7.23-7.39(m,6H),4.95-5.05(m,2H),3.87(br s,1H),3.01-3.11(m,2H),1.55-1.65(m,1H),1.35-1.45(m,1H),1.15-1.25(m,1H),0.79-0.87(m,6H).m / z(ESI):319.0(M+H) + .

[0288] Step 4: (S)-4-methyl-1-(thiazole-2-yl)pentan-2-amine A mixture of benzyl(S)-(4-methyl-1-(thiazole-2-yl)pentan-2-yl)carbamate (1.8 g, 5.65 mmol) and hydrobromic acid in AcOH (15 mL, 91 mmol) was stirred at rt for 1 hour. The reaction mixture was then quenched with a saturated aqueous solution of NaHCO3 and extracted with 15% MeOH in DCM (3 × 100 mL). The combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain (S)-4-methyl-1-(thiazole-2-yl)pentan-2-amine (1.0 g, 3.91 mmol, yield 69%) as a pale yellow oil. m / z(ESI): 185.1(M+H) + .

[0289] Amine 7: (S)-4-methyl-1-(5-methyl-1,2,4-oxadiazole-3-yl)pentan-2-amine hydrochloride [ka] Step 1: tert-butyl(S)-(1-cyano-4-methylpentan-2-yl)carbamate To a suspension of tert-butyl(S)-(1-amino-5-methyl-1-oxohexane-3-yl)carbamate (3.0 g, 12.3 mmol) and pyridine (2.0 mL, 24.6 mmol) in DCM (100 mL), TFAA (3.47 mL, 24.6 mmol) was added, and the reaction mixture was stirred under rt for 2 hours. Next, the reaction mixture was diluted with water and extracted with DCM. The combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified using a Redi-Sep pre-packed silica gel column (40 g) eluted with a gradient of 0-15% siRNA in hexane to obtain tert-butyl(S)-(1-cyano-4-methylpentan-2-yl)carbamate (2.4 g, 10.6 mmol, yield 86%). 1¹H NMR (400MHz, chloroform-d): δ ppm 4.63 (br s,1H), 3.91 (m,1H), 2.79 (m,1H), 2.49 (m,1H), 1.63-1.76 (m,1H), 1.58 (m,1H), 1.46 (s,9H), 1.40 (m,1H), 0.96 (m,6H). m / z (ESI): 127.2 (M-Boc+H) + .

[0290] Step 2: tert-butyl(S)-(4-methyl-1-(5-methyl-1,2,4-oxadiazole-3-yl)pentan-2-yl)carbamate A solution of tert-butyl(S)-(1-cyano-4-methylpentan-2-yl)carbamate (2.4 g, 10.6 mmol) and hydroxylamine (5 mL, 50% in water) in EtOH (50 mL) was heated at 60°C for 16 hours. The reaction mixture was then concentrated under reduced pressure. The residue was diluted with water and extracted with ethyl acetate. The combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain tert-butyl(S)-(1-(hydroxyamino)-1-imino-5-methylhexane-3-yl)carbamate, which was then transferred to the next step without purification. m / z(ESI): 260.1(M+H) + .

[0291] A solution of (S)-3-((tert-butoxycarbonyl)amino)-5-methylhexanimidoperoxoic acid (1.5 g, 5.76 mmol) and Ac2O (0.60 mL, 6.34 mmol) in 1,4-dioxane (3 mL) was heated in a microwave reactor (Personal Chemistry, Biotage AB, Inc., Upsala, Sweden) at 150 °C for 60 minutes. Subsequently, the reaction mixture was concentrated under reduced pressure, and the residue was purified using a Redi-Sep pre-packed silica gel column (40 g) eluting with a gradient of 0-15% ethyl acetate in PE to obtain tert-butyl(S)-(4-methyl-1-(3-methyl-1,2,4-oxadiazole-5-yl)pentan-2-yl)carbamate (1.0 g, 3.53 mmol, yield 61%).1 ¹H NMR (400MHz, chloroform-d): δ ppm 4.85(m,1H), 4.04-4.17(m,1H), 2.89(m,2H), 2.59(s,3H), 1.71(m,1H), 1.44(s,9H), 1.33-1.40(m,1H), 1.27(m,1H), 0.93(m,6H). m / z (ESI): 184.1(M-Boc+H) + .

[0292] Step 3: (S)-4-methyl-1-(5-methyl-1,2,4-oxadiazole-3-yl)pentan-2-amine hydrochloride To a solution of tert-butyl(S)-(4-methyl-1-(3-methyl-1,2,4-oxadiazole-5-yl)pentan-2-yl)carbamate (0.50 g, 1.76 mmol) in 1,4-dioxane (5 mL), 4.0 M HCl (5 mL) was added to the dioxane, and the reaction mixture was stirred under rt for 16 hours. Subsequently, the reaction mixture was concentrated under reduced pressure and triturated with Et2O to obtain (S)-4-methyl-1-(5-methyl-1,2,4-oxadiazole-3-yl)pentan-2-amine hydrochloride, which was used directly in the next step. m / z(ESI): 184.1(M+H) + .

[0293] Amine 8: (S)-4-methyl-1-(1H-1,2,4-triazol-1-yl)pentan-2-amine hydrochloride [ka] Step 1: (S)-2-((tert-butoxycarbonyl)amino)-4-methylpentylmethanesulfonate A solution of tert-butyl(S)-(1-hydroxy-4-methylpentan-2-yl)carbamate (7.5 g, 34.5 mmol, Combi-Blocks) and TEA (14.4 mL, 104 mmol) in DCM (75 mL) was mixed with MsCl (5.93 g, 51.8 mmol, Chempure) at 0°C. The resulting mixture was stirred at rt for 1 hour, then diluted with ice-cold water and extracted with DCM. The combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain (S)-2-((tert-butoxycarbonyl)amino)-4-methylpentylmethanesulfonate (10.5 g) as a grayish-white solid, which was used directly in the next step. m / z(ESI): 196.1(M-Boc+H) + .

[0294] Step 2: tert-butyl(S)-(4-methyl-1-(1H-1,2,4-triazol-1-yl)pentan-2-yl)carbamate To a solution of (S)-2-((tert-butoxycarbonyl)amino)-4-methylpentylmethanesulfonate (1.3 g, 4.40 mmol) in DMF (10 mL), Cs2CO3 (2.15 g, 6.60 mmol) and 1H-1,2,4-triazole (0.35 g, 5.07 mmol, Combi-Blocks) were added, and the resulting mixture was stirred at 80°C for 6 hours. Subsequently, the reaction mixture was diluted with ice-cold water, and the precipitated solid was filtered and dried to obtain tert-butyl(S)-(4-methyl-1-(1H-1,2,4-triazole-1-yl)pentan-2-yl)carbamate (1.1 g, 4.10 mmol, yield 93%) as a white solid. 1 H NMR(400MHz,DMSO-d6):δ ppm 8.36(s,1H),7.95(d,J=10.8Hz,1H),6.78(d,J=9.0Hz,1H),3.98-4.22(m,2H),3.72-3.92(m ,1H),1.54-1.70(m,1H),1.36(s,9H),1.08(m,1H),0.78-0.94(m,6H).m / z(ESI):269.1(M+H) + .

[0295] Step 3: (S)-4-methyl-1-(1H-1,2,4-triazol-1-yl)pentan-2-amine hydrochloride To a solution of tert-butyl(S)-(4-methyl-1-(1H-1,2,4-triazole-1-yl)pentan-2-yl)carbamate (1.1 g, 4.10 mmol) in DCM (5 mL), 4.0 M HCl (5.1 mL, 20.4 mmol) in dioxane was added, and the reaction mixture was stirred under rt for 2 hours. Subsequently, the reaction mixture was concentrated under reduced pressure and triturated with Et2O to obtain (S)-4-methyl-1-(1H-1,2,4-triazole-1-yl)pentan-2-amine hydrochloride (0.5 g, 2.44 mmol, yield 59%) as a white solid, which was then transferred to the next step without further purification. m / z(ESI): 169.2(M+H) + .

[0296] [Table 23]

[0297] Amine 11: (S)-1-(isoxazole-3-yl)-4-methylpentan-2-amine hydrochloride [ka] Step 1: tert-butyl(S)-(1-hydroxy-5-methylhexane-3-yl)carbamate To a solution of (S)-3-((tert-butoxycarbonyl)amino)-5-methylhexanoic acid (6.0 g, 24.5 mmol) in THF (90 mL), borane-THF complex (1 M in THF) (73.4 mL, 73.4 mmol, Symax) was added dropwise at 0°C, and the reaction mixture was stirred in rt for 16 hours. Subsequently, the reaction mixture was quenched with 1N HCl at 0°C and extracted with ethyl acetate. The combined organic extracts were washed with brine, separated, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain tert-butyl(S)-(1-hydroxy-5-methylhexane-3-yl)carbamate (4.0 g, 17.3 mmol, yield 71%), which was then carried to the next step without purification.1 H NMR(400MHz,DMSO-d6):δ ppm 6.52(s,2H),4.30(m,1H),3.38(m,4H),1.35-1.53(m,9H),1.25-1.35(m,2H),1.11(m,1H),0.72-0.96(m,6H).m / z(ESI):132.3(M-Boc+H) + .

[0298] Step 2: tert-butyl(S)-(5-methyl-1-oxohexane-3-yl)carbamate To a solution of tert-butyl(S)-(1-hydroxy-5-methylhexane-3-yl)carbamate (2.0 g, 8.65 mmol) in DCM (20 mL), DMP (5.50 g, 12.97 mmol, Spectrochem) was added, and the resulting solution was stirred at rt for 1 hour. Subsequently, the reaction mixture was quenched with a saturated aqueous solution of sodium carbonate and extracted with DCM. The combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified using a Redi-Sep pre-packed silica gel column (40 g) eluting with a gradient of 0-10% siRNA in hexane to obtain tert-butyl(S)-(5-methyl-1-oxohexane-3-yl)carbamate (0.55 g, 2.40 mmol, yield 28%) as a white solid. 1 ¹H NMR (400MHz, chloroform-d): δ ppm 9.78 (s,1H), 4.58-4.64 (m,1H), 4.12 (m,1H), 2.51-2.69 (m,2H), 1.46 (m,2H), 1.44 (s,9H), 1.25-1.36 (m,1H), 0.95 (m,6H). m / z (ESI): 129.0 (M-Boc+H) + .

[0299] Step 3: tert-butyl(S,E)-(1-(hydroxyimino)-5-methylhexane-3-yl)carbamate To a solution of tert-butyl(S)-(5-methyl-1-oxohexane-3-yl)carbamate (0.55 g, 2.40 mmol) in DCM (11 mL), small amounts of TEA (0.33 mL, 2.40 mmol) and hydroxylamine hydrochloride (0.20 g, 2.88 mmol) were added at 0°C. The reaction mixture was stirred at rt for 1 hour, and then concentrated under reduced pressure to obtain tert-butyl(S,E)-(1-(hydroxyimino)-5-methylhexane-3-yl)carbamate (0.80 g) as a white solid, which was then carried to the next step without purification. 1 H NMR(400MHz,DMSO-d6):δ ppm 10.42(s,1H),7.23(t,J=6.2Hz,1H),6.73(m,1H),3.61(m,1H),2.07-2.25(m,2H),1 .51-1.62(m,2H),1.38(s,9H),1.13(m,1H),0.85(m,6H).m / z(ESI):145.2(M-Boc+H) + .

[0300] Step 4: tert-butyl(S,Z)-(1-chloro-1-(hydroxyimino)-5-methylhexane-3-yl)carbamate To a solution of tert-butyl(S,E)-(1-(hydroxyimino)-5-methylhexane-3-yl)carbamate (0.80 g, 3.27 mmol) in DMF (5 mL), NCS (0.53 g, 3.93 mmol, Avra) was added, and the reaction mixture was heated at 50°C for 2 hours. Subsequently, the reaction mixture was cooled to rt, quenched with ice water, and extracted with ethyl acetate. The combined organic extracts were washed with brine, separated, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain tert-butyl(S,Z)-(1-chloro-1-(hydroxyimino)-5-methylhexane-3-yl)carbamate (0.85 g), which was then directly transferred to the next step. m / z(ESI): 179.1(M-Boc+H) + .

[0301] Step 5: tert-butyl(S)-(4-methyl-1-(5-(trimethylsilyl)isoxazole-3-yl)pentan-2-yl)carbamate A solution of tert-butyl(S,Z)-(1-chloro-1-(hydroxyimino)-5-methylhexane-3-yl)carbamate (0.85 g, 3.05 mmol), trimethylsilylacetylene (0.428 mL, 3.05 mmol), and TEA (0.29 mL, 3.05 mmol) in DCM (4 mL) was stirred at rt for 16 hours. The reaction mixture was then diluted with water and extracted with DCM. The combined organic extract was washed with brine, separated, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified using a Redi-Sep pre-packed silica gel column (12 g) eluted with 0-5% ethyl acetate in hexane to obtain tert-butyl(S)-(4-methyl-1-(5-(trimethylsilyl)isoxazole-3-yl)pentan-2-yl)carbamate (0.50 g, 1.47 mmol, yield 48%) as a colorless liquid. 1 H NMR(400MHz,DMSO-d6):δ ppm 6.73(d,J=9.3Hz,1H),6.55(d,J=1.9Hz,1H),3.78(m,1H),2.61-2.78(m,2H),1.60( m,1H),1.32(m,10H),1.19(m,1H),0.85(m,6H),0.28(s,9H).m / z(ESI):341.0(M+H) + .

[0302] Step 6: tert-butyl(S)-(1-(isoxazole-3-yl)-4-methylpentan-2-yl)carbamate A solution of tert-butyl(S)-(4-methyl-1-(5-(trimethylsilyl)isoxazole-3-yl)pentan-2-yl)carbamate (0.50 g, 1.47 mmol) and potassium carbonate (0.41 g, 2.94 mmol) in MeOH (10 mL) was stirred at rt for 1 hour. Next, the reaction mixture was diluted with water and extracted by DCM. The combined organic extracts were washed with brine, separated, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified using a Redi-Sep pre-packed silica gel column (12 g) eluted with 0-15% ELISA in hexane to obtain tert-butyl(S)-(1-(isoxazole-3-yl)-4-methylpentan-2-yl)carbamate (0.32 g, 1.19 mmol, yield 81%) as a white solid. 1 H NMR(400MHz,DMSO-d6):δ ppm 8.77(s,1H),6.74(d,J=8.4Hz,1H),3.75(m,1H),2.67-2.74(m,2H),1.58-1.61(m,1 H),1.34(m,10H),1.12-1.18(m,1H),0.83-0.86(m,6H).m / z(ESI):169.1(M-Boc+H) + .

[0303] Step 7: (S)-1-(isoxazole-3-yl)-4-methylpentan-2-amine hydrochloride To a solution of tert-butyl(S)-(1-(isoxazole-3-yl)-4-methylpentan-2-yl)carbamate (0.32 g, 1.19 mmol) in 1,4-dioxane (3.2 mL), HCl (4 M in dioxane) (1.5 mL, 6.0 mmol) was added, and the reaction mixture was stirred under rt for 1 hour. Subsequently, the reaction mixture was concentrated under reduced pressure and triturated with Et2O to obtain (S)-1-(isoxazole-3-yl)-4-methylpentan-2-amine hydrochloride (0.35 g) as a white solid. 1H NMR(400MHz,DMSO-d6):δ ppm 8.91(s,1H),8.22(br s,3H),6.64(s,1H),3.46-3.54(m,1H),3.07(dd,J=14.9,5.5Hz,1H),2.97(dd,J=14.9, 7.6Hz,1H),1.74(m,1H),1.50(m,1H),1.28(m,1H),0.84(m,6H).m / z(ESI):169.2(M+H) + .

[0304] Amine 12: (S)-4-methyl-1-(3-methylisoxazol-5-yl)pentan-2-amine hydrochloride [ka] Step 1: tert-butyl(S)-(6-methylhept-1-in-4-yl)carbamate To a solution of tert-butyl(S)-(5-methyl-1-oxohexane-3-yl)carbamate (1.9 g, 8.29 mmol) and potassium carbonate (2.29 g, 16.6 mmol) in MeOH (20 mL), dimethyl(1-diazo-2-oxopropyl)phosphonate (1.91 g, 9.94 mmol, Chemimpex) was added dropwise at 0°C. The reaction mixture was slowly heated to rt and stirred for 16 hours. Subsequently, the reaction mixture was quenched with ice-cold water and extracted with Et2O. The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified using a Redi-Sep pre-packed silica gel column (40 g) eluted with an 8-10% siRNA gradient in hexane to obtain tert-butyl(S)-(6-methylhepto-1-in-4-yl)carbamate (0.70 g, 3.11 mmol, yield 37%) as a colorless liquid. m / z(ESI): 126.2(M-Boc+H) + .

[0305] Step 2: tert-butyl(S)-(4-methyl-1-(3-methylisoxazol-5-yl)pentan-2-yl)carbamate To a solution of acetaldehyde (1.37 g, 31.1 mmol) in tert-butanol (15 mL) and water (15 mL), hydroxylamine hydrochloride (2.16 g, 31.1 mmol) and NaOH (1.24 g, 31.1 mmol) were added. After stirring the above solution for 30 minutes, chloramine-T trihydrate (8.75 g, 31.1 mmol), copper(II) sulfate pentahydrate (0.39 g, 1.55 mmol), copper (0.04 g, 0.621 mmol), and tert-butyl(S)-(6-methylhepto-1-in-4-yl)carbamate (0.70 g, 3.11 mmol) were added to the reaction mixture. The resulting mixture was stirred in rt for 20 hours, then diluted with saturated NH4Cl and extracted with phenylethylamine. The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified using a Redi-Sep pre-packed silica gel column (40 g) eluted with a 20-30% siRNA gradient in hexane to obtain tert-butyl(S)-(4-methyl-1-(3-methylisoxazole-5-yl)pentan-2-yl)carbamate (0.32 g, 1.13 mmol, yield 36%) as a pale yellow oil. m / z(ESI): 283.0(M+H) + .

[0306] Step 3: (S)-4-methyl-1-(3-methylisoxazole-5-yl)pentan-2-amine hydrochloride To a solution of tert-butyl(S)-(4-methyl-1-(3-methylisoxazole-5-yl)pentan-2-yl)carbamate (0.32 g, 1.13 mmol) in DCM (1 mL), HCl (4 M in dioxane) (0.85 mL, 3.40 mmol) was added, and the reaction mixture was stirred under rt for 3 hours. Subsequently, the reaction mixture was concentrated under reduced pressure and triturated with Et2O to obtain (S)-4-methyl-1-(3-methylisoxazole-5-yl)pentan-2-amine hydrochloride (0.20 g, 0.91 mmol, yield 81%) as a white solid. m / z(ESI): 183.1(M+H) + .

[0307] Amine 13: (S)-3-amino-N-methyl-4-phenylbutanamide hydrochloride [ka] Step 1: tert-butyl(S)-(4-(methylamino)-4-oxo-1-phenylbutan-2-yl)carbamate A solution of (S)-3-((tert-butoxycarbonyl)amino)-4-phenylbutanoic acid (1.5 g, 5.37 mmol, Combi-Blocks), methaneamine hydrochloride (0.471 g, 6.98 mmol), DIPEA (2.81 mL, 16.1 mmol), EDC-HCl (1.24 g, 6.44 mmol), and HOBt (0.82 g, 5.37 mmol) in DMF (7.5 mL) was stirred at rt for 16 hours. Subsequently, the reaction mixture was treated with ice-cold water, and the precipitated solid was filtered and dried to obtain tert-butyl(S)-(4-(methylamino)-4-oxo-1-phenylbutan-2-yl)carbamate (0.70 g, 2.39 mmol, yield 45%) as a white solid. 1 H NMR(400MHz,DMSO-d6):δ ppm 7.71(d,J=4.0Hz,1H),7.26(m,2H),7.17(m,3H),6.68(d,J=8.4Hz 1H),3.93(m,1H),2.67(m,2H),2.54(s,3H),2.20(m,2H),1.30(s,9H).m / z(ESI)m / z:293.0(M+H) + .

[0308] Step 2: (S)-3-amino-N-methyl-4-phenylbutanamide hydrochloride To a solution of tert-butyl(S)-(4-(methylamino)-4-oxo-1-phenylbutan-2-yl)carbamate (0.70 g, 2.39 mmol) in DCM (5 mL), HCl (4 M in dioxane) (1.0 mL, 4.0 mmol) was added dropwise at 0°C. The resulting reaction mixture was stirred at rt for 6 hours, then concentrated under reduced pressure and triturated with Et2O to obtain (S)-3-amino-N-methyl-4-phenylbutanamide hydrochloride (0.50 g, 2.19 mmol, yield 91%) as a grayish-white solid. 1 H NMR(400MHz,DMSO-d6)δ ppm 8.07-8.13(m,3H),7.31-7.40(m,2H),7.20-7.31(m,3H),3.65(m,1H),3. 17(s,1H),2.77(m,1H),2.56(s,3H),2.37(m,2H).m / z(ESI):193.1(M+H) + .

[0309] Amine 14: (S)-4-methyl-1-(5-methyl-1,3,4-thiadiazole-2-yl)pentan-2-amine hydrochloride [ka] Step 1: tert-butyl(S)-(1-(2-acetylhydrazineyl)-5-methyl-1-oxohexane-3-yl)carbamate A solution of (S)-3-((tert-butoxycarbonyl)amino)-5-methylhexanoic acid (5.0 g, 20.4 mmol) in DMF (50 mL), along with EDC.HCl (4.69 g, 24.5 mmol, Chempure), HOBt (3.75 g, 24.5 mmol, Chempure), acetohydrazide (1.81 g, 24.5 mmol, Combi-Blocks), and DIPEA (7.1 mL, 40.8 mmol) was stirred at rt for 16 hours. The reaction mixture was then diluted with water and extracted with ethyl acetate. The residue was triturated with 20% ethyl acetate in PE to obtain tert-butyl(S)-(1-(2-acetylhydrazineyl)-5-methyl-1-oxohexane-3-yl)carbamate (4.5 g, 14.9 mmol, yield 73%) as a white solid. m / z(ESI):202.1(M-Boc+H) + .

[0310] Step 2: tert-butyl(S)-(4-methyl-1-(5-methyl-1,3,4-thiadiazole-2-yl)pentan-2-yl)carbamate A solution of tert-butyl(S)-(4-methyl-1-(5-methyl-1,3,4-thiadiazole-2-yl)pentan-2-yl)carbamate (0.80 g, 2.67 mmol) and Lawson's reagent (0.74 g, 1.83 mmol) in 1,4-dioxane (10 mL) was heated at 100°C for 2 hours. The reaction mixture was then stirred in rt for 16 hours, diluted with water, and extracted with DCM. The combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain tert-butyl(S)-(4-methyl-1-(5-methyl-1,3,4-thiadiazole-2-yl)pentan-2-yl)carbamate (0.80 g, 2.67 mmol, yield 81%) as a grayish-white solid. m / z(ESI): 300.2(M+H) + .

[0311] Step 3: (S)-4-methyl-1-(5-methyl-1,3,4-thiadiazole-2-yl)pentan-2-amine hydrochloride To a solution of tert-butyl(S)-(4-methyl-1-(5-methyl-1,3,4-thiadiazole-2-yl)pentan-2-yl)carbamate (0.80 g, 2.67 mmol)) in 1,4-dioxane (2 mL), HCl (4.0 M solution in dioxane) (4 mL, 16.0 mmol) was added dropwise at 0°C, and the reaction mixture was stirred under rt for 2 hours. Subsequently, the reaction mixture was concentrated under reduced pressure and triturated with Et2O to obtain (S)-4-methyl-1-(5-methyl-1,3,4-thiadiazole-2-yl)pentan-2-amine hydrochloride (0.65 g), which was then carried to the next step without purification. 1 H NMR(400MHz,DMSO-d6):δ ppm 8.31-8.35(brs,3H),6.68(br s,3H),3.58(m,1H),3.34-3.49(m,2H),2.71(s,3H),1.76(m,1H),1.56(m,1H),1.36(m,1H),0.85(m,6H).m / z(ESI):200.1(M+H) + .

[0312] Amine 15: (S)-4-methyl-1-(5-methyl-1,3,4-oxadiazole-2-yl)pentan-2-amine hydrochloride [ka] Step 1: tert-butyl(S)-(4-methyl-1-(5-methyl-1,3,4-oxadiazole-2-yl)pentan-2-yl)carbamate A solution of tert-butyl(S)-(1-hydrazineyl-5-methyl-1-oxohexane-3-yl)carbamate (0.5 g, 1.93 mmol) in triethyl orthoacetate (10 mL, 54.2 mmol, Avra) was heated at 130°C for 16 hours. The reaction mixture was then concentrated under reduced pressure, the residue was treated with an aqueous solution of K2CO3, and extracted with ELISA. The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain tert-butyl(S)-(4-methyl-1-(5-methyl-1,3,4-oxadiazole-2-yl)pentan-2-yl)carbamate (0.80 g), which was then transferred to the next step without purification. m / z(ESI): 284.0(M+H) + .

[0313] Step 2: (S)-4-methyl-1-(5-methyl-1,3,4-oxadiazole-2-yl)pentan-2-amine hydrochloride To a solution of tert-butyl(S)-(4-methyl-1-(5-methyl-1,3,4-oxadiazole-2-yl)pentan-2-yl)carbamate (0.80 g, 2.82 mmol)) in 1,4-dioxane (2 mL), HCl in dioxane (4.0 M solution in dioxane) (2 mL, 8.0 mmol) was added dropwise, and the reaction mixture was stirred under rt for 2 hours. Subsequently, the reaction mixture was concentrated under reduced pressure and triturated with Et2O to obtain (S)-4-methyl-1-(5-methyl-1,3,4-oxadiazole-2-yl)pentan-2-amine hydrochloride. m / z(ESI): 184.1(M+H) + .

[0314] Amine 16: (S)-4-methyl-1-(5-methylisoxazol-3-yl)pentan-2-amine hydrochloride [ka] Step 1: tert-butyl(S)-(4-methyl-1-(5-methylisoxazol-3-yl)pentan-2-yl)carbamate To a solution of tert-butyl(S)-(5-methyl-1-oxohexane-3-yl)carbamate (1.1 g, 4.80 mmol) and TEA (1.34 mL, 9.59 mmol) in DCM (20 mL), hydroxylamine hydrochloride (0.67 g, 9.59 mmol) was added in small increments at 0°C. The reaction mixture was stirred at rt for 1 hour, then concentrated under reduced pressure to obtain tert-butyl(S,E)-(1-(hydroxyimino)-5-methylhexane-3-yl)carbamate (1.3 g) as a white solid, which was then transferred to the next step without purification. m / z(ESI): 145.2(M-Boc+H) + .

[0315] To a solution of tert-butyl(S,E)-(1-(hydroxyimino)-5-methylhexane-3-yl)carbamate (1.3g) in tert-butanol (10mL) and water (10mL), chloramine-T trihydrate (2.70g, 9.59 mmol), copper (0.03g, 0.48 mmol), copper(II) sulfate pentahydrate (0.12g, 0.48 mmol), and propane-1-yne (5% in THF) (19.2g, 24.0 mmol) were added, and the resulting mixture was stirred at rt for 16 hours. Subsequently, the reaction mixture was diluted with saturated NH4Cl aqueous solution and extracted with Et2O. The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified using a Redi-Sep pre-packed silica gel column (12 g) eluted with a 15-20% siRNA gradient in hexane to obtain tert-butyl(S)-(4-methyl-1-(5-methylisoxazole-3-yl)pentan-2-yl)carbamate (0.50 g, 1.77 mmol, yield 37%) as a pale yellow oil. m / z(ESI): 283.0(M+H) + .

[0316] Step 2: (S)-4-methyl-1-(5-methylisoxazol-3-yl)pentan-2-amine hydrochloride To a solution of tert-butyl(S)-(4-methyl-1-(5-methylisoxazole-3-yl)pentan-2-yl)carbamate (0.50 g, 1.77 mmol) in DCM (1 mL), HCl (4 M in dioxane) (1.33 mL, 5.3 mmol) was added, and the reaction mixture was stirred under rt for 3 hours. Subsequently, the reaction mixture was concentrated under reduced pressure and triturated with Et2O to obtain (S)-4-methyl-1-(5-methylisoxazole-3-yl)pentan-2-amine hydrochloride (0.35 g, 1.60 mmol, 90% yield) as a white solid. m / z(ESI): 183.1(M+H) + .

[0317] Amine 17: (S)-1-(1H-imidazole-2-yl)-4-methylpentan-2-amine hydrochloride [ka] Step 1: tert-butyl(S)-(1-(1H-imidazole-2-yl)-4-methylpentan-2-yl)carbamate A solution of tert-butyl(S)-(5-methyl-1-oxohexane-3-yl)carbamate (0.80 g, 3.49 mmol), oxalaldehyde (0.66 g, 4.54 mmol), and 7M ammonia (1.30 mL, 9.07 mmol) in MeOH (0.8 mL) was stirred in a sealed tube of rt for 16 hours. The reaction mixture was then concentrated under reduced pressure. The residue was dissolved in water and extracted with ELISA. The combined organic extracts were washed with brine, separated, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was triturated with Et2O to obtain tert-butyl(S)-(1-(1H-imidazole-2-yl)-4-methylpentan-2-yl)carbamate (0.30 g, 1.12 mmol, yield 32%) as a grayish-white solid. 1H NMR(300MHz,DMSO-d6):δ ppm 11.69(s,1H),6.85(s,2H),6.69(d,J=9.0Hz,1H),3.83(m,1H),2.65(m,2H),1. 55(m,1H),1.20-1.46(m,10H),1.04(m,1H),0.80(m,6H).m / z(ESI):268.0(M+H) + .

[0318] Step 2: (S)-1-(1H-imidazole-2-yl)-4-methylpentan-2-amine hydrochloride To a solution of tert-butyl(S)-(1-(1H-imidazole-2-yl)-4-methylpentan-2-yl)carbamate (0.30 g, 1.12 mmol) in 1,4-dioxane (3 mL), HCl (4 M in dioxane) (1.40 mL, 5.61 mmol) was added, and the reaction mixture was stirred under rt for 4 hours. Subsequently, the reaction mixture was concentrated under reduced pressure and triturated with Et2O to obtain (S)-1-(1H-imidazole-2-yl)-4-methylpentan-2-amine hydrochloride (0.50 g). 1 H NMR(400MHz,DMSO-d6):δ ppm 8.49(s,3H),7.63(s,2H),3.85(m,1H),3.27-3.44(m,2H),1.59(m,1H),1 .51-1.68(m,1H),1.17-1.27(m,1H),0.85(m,6H).m / z(ESI):168.1(M+H) + .

[0319] Amine 18: (S)-4-methyl-1-(1H-1,2,3-triazol-5-yl)pentan-2-amine hydrochloride [ka] Step 1: tert-butyl(S)-(4-methyl-1-(1H-1,2,3-triazole-5-yl)pentan-2-yl)carbamate A solution of tert-butyl(S)-(6-methylhept-1-in-4-yl)carbamate (0.50 g, 2.22 mmol), sodium azide (0.144 g, 2.22 mmol), TBTA (0.12 g, 0.22 mmol, Combi-Blocks), L(+)-sodium ascorbate (0.088 g, 0.44 mmol), and copper(II) sulfate (0.071 g, 0.44 mmol) in THF (7.5 mL) and water (2.5 mL) was stirred at rt for 16 hours, followed by 2 hours at 50°C. The reaction mixture was then diluted with water and extracted with SiO2. The combined organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified using a Redi-Sep pre-packed silica gel column (12 g) eluted with a gradient of 0-50% siRNA in hexane to obtain tert-butyl(S)-(4-methyl-1-(1H-1,2,3-triazole-5-yl)pentan-2-yl)carbamate (0.10 g, 0.37 mmol, yield 17%) as a grayish-white solid. m / z(ESI): 269.1(M+H) + .

[0320] Step 2: (S)-4-methyl-1-(1H-1,2,3-triazol-5-yl)pentan-2-amine hydrochloride To a solution of tert-butyl(S)-(4-methyl-1-(1H-1,2,3-triazole-5-yl)pentan-2-yl)carbamate (0.10 g, 0.37 mmol) in 1,4-dioxane (1 mL), HCl (4 M in dioxane) (0.47 mL, 1.86 mmol) was added, and the reaction mixture was stirred under rt for 3 hours. Subsequently, the reaction mixture was concentrated under reduced pressure and triturated with Et2O to obtain (S)-4-methyl-1-(1H-1,2,3-triazole-5-yl)pentan-2-amine hydrochloride (0.15 g). 1 H NMR(400MHz,DMSO-d6):δ ppm 7.94(br s,3H),7.78(s,1H),3.42(m,1H),2.96(d,J=6.3Hz,2H),1.73(m,1H),1.42(m,1H),1.29(m,1H),0.84(m,6H).m / z(ESI):169.2(M+H) + .

[0321] Analysis

[0322] Table 24

[0323] Table 25

[0324] Table 26

[0325] Table 27

[0326] Table 28

[0327] Table 29

[0328] Table 30

[0329] Table 31

[0330] Table 32

[0331] Table 33

[0332] Biological evaluation This section provides a biological evaluation of the specific examples provided herein. See Table 6.

[0333] Binding nucleotide exchange assay: Purified GDP-bound KRAS proteins (aa 1-169) containing both G12C and C118A amino acid substitutions and an N-terminal His-tag were pre-incubated for either 2 or 20 hours in assay buffer (25 mM HEPES pH 7.4, 10 mM MgCl2, and 0.01% Triton X-100) with serially diluted compounds. In all subsequent steps, DTT was added to the reaction buffer at a final concentration of 1 mM. Following the pre-incubation of the compounds, purified SOS proteins (aa 564-1049) and GTP (Roche 10106399001) were added to the assay wells and incubated for a further 30 minutes. To determine the degree of inhibition of SOS-mediated nucleotide exchange, purified GST-tagged cRAF (aa1-149), nickel chelate AlphaLISA acceptor beads (PerkinElmer AL108R), and AlphaScreen glutathione donor beads (PerkinElmer 6765302) were added to assay wells and incubated for 5 minutes. Subsequently, the assay plates were read using a plate reader that measures luminescence signals. The signal intensity of the compound-containing wells was normalized to a DMSO control, and the data were analyzed using a 4-parameter logistic regression model to determine IC50. 50 The value was calculated.

[0334] Cell viability assay: MIA PaCa-2 (human pancreatic cancer; ATCC CRL-1420) or A549 (human lung cancer; ATCC CCL-185) cells were cultured in RPMI1640 medium containing 10% fetal bovine serum and 1× penicillin / streptomycin / L-glutamine. Cells were seeded at a density of 1.67E+04 cells / mL into 384-well plates and incubated overnight at 37°C and 5% CO2. Serially diluted compounds or DMSO were added to the cells, and the plates were incubated at 37°C and 5% CO2 for 72 hours. Cell viability was measured using the CellTiter-Glo® Luminescent Cell Viability Assay kit (Promega) according to the manufacturer's protocol. The luminescence signal of the treated samples was normalized relative to the DMSO control, and the data were analyzed using a 4-parameter logistic model to determine IC50. 50 The value was calculated.

[0335] [Table 34]

[0336] [Table 35]

[0337] [Table 36]

[0338] The results shown in Table 6 were generated by the in vitro assays described above. These assays can be used to test any of the compounds described herein and to evaluate and characterize their biological activity.

[0339] Compounds that show activity in binding-exchange assays are useful in the methods provided herein (see the "Instructions for Use" section). See, for example, Lanman et al., 2020; Hong et al., 2020. The inhibitory effects of the compounds provided herein on tumor growth can be demonstrated, for example, using the following animal models.

[0340] Tumor cells are cultured, harvested, and subcutaneously transplanted into the right flank of a female athymus nude mouse. The tumor is approximately 200 mm. 3 Once the target is reached, mice are randomized to the treatment group (n=10 / group) and treatment is started (on the days shown in the graph). Tumor size and body weight are measured 2-3 times per week. Tumor volume is measured using a digital caliper and calculated as L×W×H in mm². 3 The units are expressed as follows. The statistical significance of the differences observed between growth curves can be assessed by repeated measures analysis of covariance (RMANOVA) of log-transformed tumor volume data, and the control group can be compared to the treatment group using Dunnett-adjusted multiple comparisons. For combination trials, RMANOVA can be performed on the combination group compared one-to-one with each monotherapy group.

[0341] References Caunt,CJ;Sale,MJ;Smith,PD;Cook,SJNat.Rev.Cancer 2015,15,577. Cerami,E.;Gao,J.;Dogrusoz,U.;Gross,BE;Sumer,SO;Aksoy,BA;Jacobsen,A.;Byrne,CJ;Heuer,ML;Larsson,E.;Antipin,Y.;Reva,B.;Goldberg,AP;Sander,C.;Schultz N.Cancer Discov.2012,2(5),401. Canon , J ; Rex , K ; Saiki , AY ; Mohr , C ; Cooke , K ; Bagal , D ; Gaida , K ; Holt , T ; Knutson , CG ; Koppada , N ; Lanman , BA ; Werner , J ; Rapaport , AS ; San Miguel , T ; Ortiz , R ; Osgood , T ; Sun , JR ; Zhu , X ; McCarter , JD ; Volak , LP ; Houk , BE ; Fakih , MG ; O'Neil , BH ; Price , TJ ; F alchook , GS ; Desai , J ; Kuo , J ; Govindan , R ; Hong , DS ; Ouyang , W ; Henary , H ; Arvedson , T ; Cee , VJ ; Lipford , JRNature 2019,575(7781),2 Cox,AD;Physics,SW;Kimmelman,AC;Luo,J.;There,CJNat.Rev.Drug Discov.2014,13,828. Der,CJ;Krontiris,TG;Cooper,GMProc.Natl.Acad.Sci.USA 1982,79,3637. Gao,J.;Aksoy,BA;Dogrusoz,U;Dresdner,G;Gross,B;Sumer,SO;Sun,Y;Jacobsen,A;Sinha,R;Larsson,E;Cerami,E;Sander,C;Schultz,N.Science Signaling 2013,6(269),pl1. Holderfield ,M.;Deuker,MM;McCormick,F.;McMahon,M.Nat.Rev.Cancer 2014,14,455. Hong , DS ; Fakih , MG ; Strickler , JH ; Desai , J ; Durm , GA ; Shapiro , GI ; Falchook , GS ; Price , TJ ; Sacher ,A.;Denlinger,CS;Bang,Y-J;Dy,GK;Krauss,JC;Kuboki,Y;Kuo,JC;Coveler,AL;Park,K;Kim,TW ;Barlesi , F. ;Munster , PN ;Ramalingam , SS ;Burns , TF , Meric-Bernstam , F ; T.Li,BTNEngl.J.Med.2020,383,1207. Lanman , BA ; Allen , JR ; Allen , JG ; Amegadzie , AK ; Ashton , KS ; Booker , SK ; Chen , JJ ; Chen , N ; Frohn , MG ; oodman , G ; Kopecky , DJ ; Liu , L ; Lopez , P ; Low , JD ; Ma , V ; Minatti , E ; Nguyen , TT ; Nishimura , N ; Pick rell , AJ ; Reed , AB ; Shin , Y ; Siegmund , AC ; Tamayo , NA ; Tegley , CM ; Walton , MC ; Wang , H.-L ; Wurz , RP ; Xue , M. ; AY;St.Miguel,T;Volak,LP;Wang,KH;Whittington,DA;Zech,SG;Lipford,JR;Cee,VJJMed.Chem.2020,63,52. Malumbres,M.;Barbacid,M.Nat.Rev.Cancer 2003,3,459. O'Bryan,JPPharmacol.Res.2019,139,503. Ostrem, JM; Peters, U.; Sos, ML; Wells, JA; Shokat, KMNature 2013,503,548. Simanshu,DK;Nissley,DV;McCormick,F.Cell 2017,170,17. Sridhar, SS; Seymour, L.; Shepherd, FALancet Oncol. 2003, 4, 397. Vojtek,AB;Der,CJJBiol.Chem.1998,273,19925.

[0342] All references cited herein, such as scientific papers or patent application publications, are incorporated herein by reference in their entirety for all purposes to the same extent that each reference is specifically and individually indicated as being incorporated in its entirety for all purposes.

Claims

1. Compound of formula I 【Chemistry 1】 or a pharmaceutically acceptable salt thereof, in the formula, R １ Each instance of appearance is independent of H and C. １～４ Alkoxy, -(CH ２ )-C １～４ Dialkylamino, aziridine-1-ylmethyl, azetidine-1-ylmethyl, pyrrolidine-1-ylmethyl, piperidine-1-ylmethyl, or morpholine-1-ylmethyl, R ２ is H, halogen, -CN, C １～４ alkyl, C １～４ haloalkyl, -CH ２ CN, -CH ２ OH, C １～４ alkoxy, or C １～４ haloalkoxy, and Selectively, one R １ and R ２ These, along with the carbon atoms to which they are bonded, 【Chemistry 2】 Forming a base, R ３ Each instance is independently represented as H, halogen, CN, OH, -CH ２ OH, C １～４ Alkyl, C １～４ Haloalkyl, -CH ２ CN, or C １～４ It is an alkoxy compound, and has two substituents R bonded to the same carbon atom. ３ C is optionally selected together with the carbon atom. ３～６ Forming a cycloalkyl or carbonyl group, A is independent each time it appears, CR ３ R ３ Or absent, R ４ is, Z １ -CH(Z ２ -R ５ ) - CH ２ -R ６ And, Z １ is O, NH, N(C １～４ Alkyl), or CH ２ And, Z ２ is absent or CH ２ And, R ５ C １～４ Alkyl, C １～４ Haloalkyl, C ３～６ Cycloalkyl, C ３～６ It is a heterocycloalkyl, phenyl, or 5-6 member heteroaryl, The phenyl may optionally contain a halogen, -CN, or C. １～３ Alkyl, C １～３ Haloalkyl, C １～３ Alkoxy and C １～３ Substituted with 1 to 3 substituents selected from haloalkoxys, The heteroaryl can be optionally composed of -CN,C １～４ Alkyl, C １～４ Haloalkyl, C １～４ Alkoxy and C １～４ Substituted with 1 to 3 substituents selected from haloalkoxys, R ６ is -CO(NR ７ R ７ ), phenyl, 5,5-dimethyl-3,5-dihydro-4H-imidazole-4-on-2-yl, or a 5-6 membered heteroaryl, wherein the heteroaryl is optionally -CN,C １～４ Alkyl, C １～４ Haloalkyl, C １～４ Alkoxy and C １～４ Substituted with 1 to 3 substituents selected from haloalkoxys, R ７ Each instance is independently H or C １～４ It is alkyl, X １ CR ８ or N, X ２ is CH or N, X ３ is C or N, X ４ is C or N, R ８ H, halogen, CN, C １～４ Alkyl, C １～４ Haloalkyl, C １～４ Alkoxy, C １～４ Haloalkoxy, C ３～５ Cycloalkyl, or C ３～５ It is a cyclohaloalkyl, B, together with the atom to which it is bonded, forms a 4- to 7-membered fully saturated, fully unsaturated, or partially unsaturated carbon ring or heterocyclic system. The aforementioned heterocyclic system comprises 1 to 5 heteroatoms selected from N, O, and S. The aforementioned ring system optionally comprises 1 to 5 substituents R ９ Replaced by, R ９ Each instance is independently of halogen, OH, -CN, and -NH. ２ , C(=O)C １～６ Alkyl, C １～６ Alkyl, C １～６ Haloalkyl, C １～４ Alkoxy, C １～４ Haloalkoxy, C ３～５ Cycloalkyl, C ３～５ It is a cyclohaloalkyl, phenyl, or 5-6 member heteroaryl, Said C １～６ Alkyl is optionally -CO(C) １～４ Alkylamino) or -CO(C １～４ Substituted with dialkylamino, The phenyl is optionally substituted with one to three independently selected halogens. The heteroaryl can optionally be a halogen, C １～４ Alkyl and C １～４ A compound of formula I, or a pharmaceutically acceptable salt thereof, substituted with one to three substituents selected from haloalkyl groups.

2. The compound according to Claim 1, or a pharmaceutically acceptable salt thereof, wherein R3 is not -CN.

3. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R5 is not pyrazole-3-yl, 2-methyl.

4. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein Z 2 is absent and R 5 is 2-cyanophenyl.

5. B is, 【Transformation 3】 Not the compound described in any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof.

6. The aforementioned compound, (3R)-3-(3-cyanophenyl)-N-methyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)propanamide; (3S)-3-((2-((7S)-7-(hydroxymethyl)-2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; 1-(6-(4-((((2S)-4-methyl-1-(1H-pyrazole-3-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; (3S)-3-((2-(8-cyano-2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-3-(2-cyanophenyl)-N-methyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)propanamide; (3R)-3-(2-cyanophenyl)-N-methyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)propanamide; (3S)-N,5-dimethyl-3-((8-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7,8-dihydro-6H-pyrimido[5,4-b][1,4]oxazine-4-yl)amino)hexanamide; (3S)-N,5-dimethyl-3-((7-(2-propanyl)-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7H-pyrrolo[2,3-d]pyrimidine-4-yl)amino)hexanamide; or The compound according to claim 1, which is not (3S)-N,5-dimethyl-3-((7-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7H-purine-6-yl)amino)hexanamide, or a pharmaceutically acceptable salt thereof.

7. Each R １ A compound according to any one of claims 1 to 6, wherein is H, or a pharmaceutically acceptable salt thereof.

8. One R １ and R ２ These, along with the carbon atoms to which they are bonded, 【Chemistry 4】 A compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, which forms a group.

9. R ２ is H, or C １～４ A compound according to any one of claims 1 to 7, which is a haloalkyl compound, or a pharmaceutically acceptable salt thereof.

10. R ２ is H, or CF ３ and is a compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof.

11. R ２ A compound according to any one of claims 1 to 7, wherein is H, or a pharmaceutically acceptable salt thereof.

12. R ３ The compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, wherein is H or a halogen.

13. R ３ The compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, wherein is H or F.

14. R ３ A compound according to any one of claims 1 to 11, wherein is H, or a pharmaceutically acceptable salt thereof.

15. One A is absent, and the other A is CR ３ R ３ The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof.

16. A compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, wherein both A's are absent. 【Request Item 17】 【Chemistry 5】 teeth, 【Transformation 6】 The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof. 【Request Item 18】 【Chemistry 7】 teeth, 【Transformation 8】 The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof. 【Request Item 19】 【Chemistry 9】 teeth, 【Chemistry 10】 The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof.

20. Z １ is NH, a compound according to any one of claims 1 to 19, or a pharmaceutically acceptable salt thereof.

21. Z ２ CH ２ The compound according to any one of claims 1 to 20, or a pharmaceutically acceptable salt thereof.

22. Z ２ The compound according to any one of claims 1 to 20, or a pharmaceutically acceptable salt thereof, is absent.

23. R ５ C １～４ A compound according to any one of claims 1 to 22, wherein the phenyl is optionally substituted with -CN, or a pharmaceutically acceptable salt thereof.

24. R ５ is -CH(CH ３ ) ２ A compound according to any one of claims 1 to 22, which is phenyl or 3-cyanophenyl, or a pharmaceutically acceptable salt thereof.

25. R ５ is -CH(CH ３ ) ２ A compound according to any one of claims 1 to 22, or a pharmaceutically acceptable salt thereof.

26. R ６ is -CO(NR ７ R ７ ), 5,5-dimethyl-3,5-dihydro-4H-imidazole-4-on-2-yl, or a 5-membered heteroaryl, wherein the heteroaryl optionally comprises 1 to 3 C １～４ Substituted with alkyl substituents, R ７ Each instance is independently H or C １～４ A compound according to any one of claims 1 to 25, which is alkyl, or a pharmaceutically acceptable salt thereof.

27. R ６ is -CO(NHR ７ ), or a five-membered heteroaryl, wherein the heteroaryl optionally contains one C １～４ Substituted with alkyl substituents, R ７ C １～４ A compound according to any one of claims 1 to 25, which is alkyl, or a pharmaceutically acceptable salt thereof.

28. R ６ is -CO(NHCH ３ ), 5,5-dimethyl-3,5-dihydro-4H-imidazole-4-on-2-yl, or a five-membered heteroaryl, wherein the heteroaryl is pyrazole, imidazole, 1,2,3-triazole, 1,2,4-triazole, 1,2-oxazole, 1,3-oxazole, 1,3,4-oxadiazole, 1,2,4-oxadiazole, 1,3-thiazole, or 1,3,4-thiadiazole, and the heteroaryl optionally contains one C １～４ A compound according to any one of claims 1 to 25, or a pharmaceutically acceptable salt thereof, substituted with an alkyl substituent.

29. R ６ is -CO(NHCH ３ A compound according to any one of claims 1 to 25, or a pharmaceutically acceptable salt thereof, wherein the heteroaryl is imidazole, 1,2-oxazole, 1,3,4-oxadiazole, 1,3,4-thiadiazole, or 1,2,3-triazole, and the heteroaryl is optionally substituted with one methyl group.

30. R ４ teeth, 【Chemistry 11】 The compound according to any one of claims 1 to 19, or a pharmaceutically acceptable salt thereof.

31. R ４ teeth, 【Chemistry 12】 The compound according to any one of claims 1 to 19, or a pharmaceutically acceptable salt thereof.

32. X １ CR ８ The compound according to any one of claims 1 to 31, or a pharmaceutically acceptable salt thereof.

33. X １ The compound according to any one of claims 1 to 31, or a pharmaceutically acceptable salt thereof, wherein is N.

34. X ２ The compound according to any one of claims 1 to 33, or a pharmaceutically acceptable salt thereof, wherein is CH.

35. X ２ The compound according to any one of claims 1 to 33, or a pharmaceutically acceptable salt thereof, wherein is N.

36. X ３ The compound according to any one of claims 1 to 35, or a pharmaceutically acceptable salt thereof, wherein C is C.

37. X ３ The compound according to any one of claims 1 to 35, or a pharmaceutically acceptable salt thereof, wherein is N.

38. X ４ The compound according to any one of claims 1 to 37, or a pharmaceutically acceptable salt thereof, wherein C is C.

39. X ４ The compound according to any one of claims 1 to 37, or a pharmaceutically acceptable salt thereof, wherein is N.

40. X １ is N, X ２ is N, X ３ C is X ４ Is it C, or X １ is N, X ２ CH is X ３ C is X ４ Is it C, or X １ is N, X ２ is N, X ３ is N, X ４ Is it C, or X １ is N, X ２ CH is X ３ C is X ４ A compound according to any one of claims 1 to 31, or a pharmaceutically acceptable salt thereof, wherein is N.

41. X １ is N, X ２ is N, X ３ C is X ４ Is it C, or X １ is N, X ２ CH is X ３ C is X ４ A compound according to any one of claims 1 to 31, wherein C is C, or a pharmaceutically acceptable salt thereof.

42. X １ is N, X ２ is N, X ３ C is X ４ A compound according to any one of claims 1 to 31, wherein C is C, or a pharmaceutically acceptable salt thereof.

43. B, along with the atom it is bonded to, 【Chemistry 13】 Form a ring system selected from, The aforementioned ring system optionally comprises 1 to 5 substituents R ９ A compound according to any one of claims 1 to 36, 38, and 40 to 42, or a pharmaceutically acceptable salt thereof, substituted with.

44. B, along with the atom it is bonded to, 【Chemistry 14】 Form a ring system selected from, The aforementioned ring system optionally comprises 1 to 5 substituents R ９ A compound according to any one of claims 1 to 36, 38, and 40 to 42, or a pharmaceutically acceptable salt thereof, substituted with.

45. B, along with the atom it is bonded to, 【Chemistry 15】 A compound according to any one of claims 1 to 36, 38, and 40 to 42, or a pharmaceutically acceptable salt thereof, which forms a ring system selected from the above.

46. The aforementioned ring system optionally includes one or two substituents R ９ Replaced by, R ９ Each instance is independently represented as halogen, -CN, and C(=O)C. １～６ Alkyl, C １～６ Alkyl, C １～６ Haloalkyl, C １～４ Alkoxy, C ３～５ A compound according to any one of claims 1 to 44, which is a cycloalkyl or a 5-6 membered heteroaryl, or a pharmaceutically acceptable salt thereof.

47. The aforementioned ring system optionally includes one or two substituents R ９ Replaced by, R ９ Each instance is independent of C １～６ Alkyl, C １～６ Haloalkyl, C １～４ Alkoxy, C ３～５ A compound according to any one of claims 1 to 44, which is a cycloalkyl or a five-membered heteroaryl, or a pharmaceutically acceptable salt thereof.

48. The aforementioned ring system optionally includes one or two substituents R ９ Replaced by, R ９ The compound according to any one of claims 1 to 44, or a pharmaceutically acceptable salt thereof, wherein each occurrence is independently Cl, -CN, acetyl, methyl, isopropyl, trifluoromethyl, methoxy, cyclopropyl, or 1,3-thiazolyl.

49. The aforementioned ring system optionally includes one or two substituents R ９ Replaced by, R ９ The compound according to any one of claims 1 to 44, or a pharmaceutically acceptable salt thereof, wherein each occurrence is independently methyl, isopropyl, trifluoromethyl, methoxy, cyclopropyl, or 1,3-thiazolyl.

50. The aforementioned compound, (S)-3-((2-(2-acryloyl-2,6-diazaspiro[3,4]octan-6-yl)-5,6,7,8-tetrahydroquinazoline-4-yl)amino)-N,5-dimethylhexaneamide; (3S)-3-((2-(8,8-difluoro-2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7-methyl-5,6,7,8-tetrahydro-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-N,5-dimethyl-3-((7-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)hexanamide; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-6,7-dihydro-5H-cyclopenta[d]pyrimidine-4-yl)amino)hexanamide; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)hexanamide; (3S)-N,5-dimethyl-3-((3-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-1-isoquinolinyl)amino)hexanamide; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)flo[3,2-d]pyrimidine-4-yl)amino)hexanamide; (3S)-N,5-dimethyl-3-(((8R)-8-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)hexanamide; (3S)-3-((6-acetyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3,4]octan-6-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidine-4-yl)amino)-N,5-dimethylhexaneamide; (3S)-N,5-dimethyl-3-((2-methyl-5-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-2H-pyrazolo[4,3-d]pyrimidine-7-yl)amino)hexanamide; (3S)-N,5-dimethyl-3-((5-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)[1,3]thiazolo[5,4-d]pyrimidine-7-yl)amino)hexanamide; 5,5-dimethyl-2-((2S)-4-methyl-2-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)pentyl)-3,5-dihydro-4H-imidazole-4-one; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,7-dihydrofl[3,4-d]pyrimidine-4-yl)amino)hexanamide; (3S)-N,5-dimethyl-3-(((8S)-8-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)hexanamide; (3S)-N,5-dimethyl-3-((7-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7H-pyrrolo[2,3-d]pyrimidine-4-yl)amino)hexanamide; (3S)-N,5-dimethyl-3-((9-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-9H-purine-6-yl)amino)hexanamide; (3S)-3-((2-(8,8-difluoro-2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; 1-(6-(4-((((2S)-4-methyl-1-(4H-1,2,4-triazole-3-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; (3S)-N,5-dimethyl-3-((7-(2-propanyl)-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)hexanamide; (3S)-3-(3-cyanophenyl)-N-methyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)propanamide; 1-(6-(4-((((2S)-4-methyl-1-(5-methyl-1,3,4-oxadiazole-2-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3,4]octan-2-yl)-2-propen-1-one; 1-(6-(4-((((2S)-1-(1H-imidazole-2-yl)-4-methyl-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; (3S)-3-((2-((7R)-7-(hydroxymethyl)-2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-N,5-dimethyl-3-((7-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)hexanamide; (3S)-3-((7-cyano-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-3-((2-(8-fluoro-2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-3-((2-(8-fluoro-2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7-methyl-5,6,7,8-tetrahydro-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-3-((7,7-dimethyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3,4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-3-((7-chloro-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7,8-dihydro-6H-pyrano[3,2-d]pyrimidine-4-yl)amino)hexanamide; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7-(trifluoromethyl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)hexanamide; (3S)-N,5-dimethyl-3-((6-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)hexanamide; (3S)-N,5-dimethyl-3-((7-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)pyrido[3,2-d]pyrimidine-4-yl)amino)hexanamide; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7-(trifluoromethyl)-4-quinazolinyl)amino)hexanamide; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7-(1,3-thiazole-2-yl)pyrido[3,2-d]pyrimidine-4-yl)amino)hexanamide; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)pyrido[3,2-d]pyrimidine-4-yl)amino)hexanamide; (3S)-3-((7-cyclopropyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)pyrido[3,2-d]pyrimidine-4-yl)amino)-N,5-dimethylhexaneamide; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(3-methyl-1,2,4-oxadiazole-5-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; (3S)-3-((7-cyclopropyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-N,5-dimethyl-3-((6-methyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)hexanamide; (3S)-3-((7-methoxy-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)pyrido[3,2-d]pyrimidine-4-yl)amino)-N,5-dimethylhexaneamide; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(1,3-thiazole-2-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(5-methyl-1,2,4-oxadiazole-3-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(1H-1,2,4-triazole-1-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(1H-pyrazole-1-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(1H-1,2,3-triazole-1-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(1,2-oxazole-3-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(3-methyl-1,2-oxazole-5-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; (3S)-3-((7,7-dimethyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3,4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)-N-methyl-4-phenylbutanamide; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(5-methyl-1,3,4-thiadiazole-2-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(5-methyl-1,3,4-oxadiazole-2-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(5-methyl-1,2-oxazole-3-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(4-((((2S)-1-(1H-imidazole-2-yl)-4-methyl-2-pentanyl)amino)-7,7-dimethyl-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(1H-1,2,3-triazole-4-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)pyrido[3,4-d]pyrimidine-4-yl)amino)hexanamide; 1-(6-(4-((((2S)-4-methyl-1-(1,3-oxazole-2-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; (3S)-N,5-dimethyl-3-((7-methyl-2-(2-(2-(trifluoromethyl)-2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)hexanamide; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7-(1,3-thiazole-2-yl)-4-quinazolinyl)amino)hexanamide; or The compound according to claim 1, which is (S)-2-((2-(2-acryloyl-2,6-diazaspiro[3.4]octan-6-yl)-5,6,7,8-tetrahydroquinazoline-4-yl)amino)-N,4-dimethylpentanamide, or a pharmaceutically acceptable salt thereof.

51. The aforementioned compound, (3S)-3-((2-(8,8-difluoro-2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7-methyl-5,6,7,8-tetrahydro-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-N,5-dimethyl-3-((7-(2-propanyl)-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)hexanamide; (3S)-3-((7,7-dimethyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3,4]octan-6-yl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7-(trifluoromethyl)-5,6,7,8-tetrahydro-4-quinazolinyl)amino)hexanamide; (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7-(1,3-thiazole-2-yl)pyrido[3,2-d]pyrimidine-4-yl)amino)hexanamide; (3S)-3-((7-cyclopropyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)pyrido[3,2-d]pyrimidine-4-yl)amino)-N,5-dimethylhexaneamide; (3S)-3-((7-cyclopropyl-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-4-quinazolinyl)amino)-N,5-dimethylhexaneamide; (3S)-3-((7-methoxy-2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)pyrido[3,2-d]pyrimidine-4-yl)amino)-N,5-dimethylhexaneamide; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(1,2-oxazole-3-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(3-methyl-1,2-oxazole-5-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(5-methyl-1,3,4-thiadiazole-2-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(5-methyl-1,3,4-oxadiazole-2-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(5-methyl-1,2-oxazole-3-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(4-((((2S)-1-(1H-imidazole-2-yl)-4-methyl-2-pentanyl)amino)-7,7-dimethyl-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; 1-(6-(7,7-dimethyl-4-(((2S)-4-methyl-1-(1H-1,2,3-triazole-4-yl)-2-pentanyl)amino)-5,6,7,8-tetrahydro-2-quinazolinyl)-2,6-diazaspiro[3.4]octan-2-yl)-2-propen-1-one; or The compound according to claim 1, which is (3S)-N,5-dimethyl-3-((2-(2-(2-propenoyl)-2,6-diazaspiro[3.4]octan-6-yl)-7-(1,3-thiazole-2-yl)-4-quinazolinyl)amino)hexaamide, or a pharmaceutically acceptable salt thereof.

52. A pharmaceutical composition comprising a compound according to any one of claims 1 to 51, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

53. A compound according to any one of claims 1 to 51, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 52, for use as a pharmaceutical agent.

54. A compound according to any one of claims 1 to 51, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 52, for use in the treatment of cancer.

55. A compound according to any one of claims 1 to 51, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 52, for use in the treatment of cancer in which one or more cells express the KRAS G12C mutant protein.

56. A compound or pharmaceutical composition for use according to claim 54 or 55, wherein the cancer is non-small cell lung cancer, small intestine cancer, appendiceal cancer, colorectal cancer, cancer of unknown primary origin, endometrial cancer, mixed-type cancer of unknown primary origin, pancreatic cancer, hepatobiliary tract cancer, small cell lung cancer, cervical cancer, germ cell cancer, ovarian cancer, gastrointestinal neuroendocrine cancer, bladder cancer, myelodysplastic / myeloproliferative neoplasm, head and neck cancer, esophageal and gastric cancer, soft tissue sarcoma, mesothelioma, thyroid cancer, leukemia, or melanoma.

57. Use of a compound according to any one of claims 1 to 51, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 52, in the preparation of a drug for treating cancer.

58. Use of a compound according to any one of claims 1 to 51, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 52, in the preparation of a drug for treating cancer, wherein one or more cells express the KRAS G12C mutant protein.

59. The use according to claim 57 or 58, wherein the cancer is non-small cell lung cancer, small intestine cancer, appendiceal cancer, colorectal cancer, cancer of unknown primary origin, endometrial cancer, mixed-type cancer of unknown primary origin, pancreatic cancer, hepatobiliary tract cancer, small cell lung cancer, cervical cancer, germ cell cancer, ovarian cancer, gastrointestinal neuroendocrine cancer, bladder cancer, myelodysplastic / myeloproliferative neoplasm, head and neck cancer, esophageal and gastric cancer, soft tissue sarcoma, mesothelioma, thyroid cancer, leukemia, or melanoma.

60. A pharmaceutical composition for use in a method of treating cancer in a subject requiring cancer treatment, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1 to 51, or a pharmaceutically acceptable salt thereof.

61. A pharmaceutical composition for use in a method of treating cancer in a subject requiring cancer treatment, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1 to 51, or a pharmaceutically acceptable salt thereof, wherein one or more cells express the KRAS G12C mutant protein.

62. The pharmaceutical composition according to claim 60 or 61, wherein the cancer is non-small cell lung cancer, small intestine cancer, appendiceal cancer, colorectal cancer, cancer of unknown primary origin, endometrial cancer, mixed-type cancer of unknown primary origin, pancreatic cancer, hepatobiliary tract cancer, small cell lung cancer, cervical cancer, germ cell cancer, ovarian cancer, gastrointestinal neuroendocrine cancer, bladder cancer, myelodysplastic / myeloproliferative neoplasm, head and neck cancer, esophageal and gastric cancer, soft tissue sarcoma, mesothelioma, thyroid cancer, leukemia, or melanoma.

63. The pharmaceutical composition according to claim 60 or 61, wherein the cancer is non-small cell lung cancer, colorectal cancer, pancreatic cancer, appendiceal cancer, endometrial cancer, esophageal cancer, cancer of unknown primary origin, ampulla cancer, gastric cancer, small intestine cancer, paranasal sinus cancer, bile duct cancer, or melanoma.

64. The pharmaceutical composition according to claim 63, wherein the cancer is non-small cell lung cancer.

65. The pharmaceutical composition according to claim 63, wherein the cancer is colorectal cancer.

66. The pharmaceutical composition according to claim 63, wherein the cancer is pancreatic cancer.

67. The pharmaceutical composition according to any one of claims 60 to 66, wherein the subject has cancer that has been determined to have one or more cells expressing the KRAS G12C mutant protein before administration of the compound or a pharmaceutically acceptable salt thereof.

68. A pharmaceutical composition according to any one of claims 60 to 67, further comprising simultaneous, individual, or sequential administration of an effective amount of a second compound, wherein the second compound is an aurora kinase A inhibitor, an AKT inhibitor, an arginase inhibitor, a CDK4 / 6 inhibitor, an ErbB family inhibitor, an ERK inhibitor, a FAK inhibitor, an FGFR inhibitor, a glutaminase inhibitor, an IGF-1R inhibitor, a KIF18A inhibitor, an MCL-1 inhibitor, a MEK inhibitor, an mTOR inhibitor, a PD-1 inhibitor, a PD-L1 inhibitor, a PI3K inhibitor, a Raf kinase inhibitor, a SHP2 inhibitor, a SOS1 inhibitor, a Src kinase inhibitor, or one or more chemotherapeutic agents.

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