PYRIDO[4,3-d]PYRIMIDINE COMPOUNDS

Abstract

To provide a novel KRAS inhibitor that can be used to treat a broader range of cancers.SOLUTION: The present invention provides compounds represented by formulas (V) to (VII), or pharmaceutically acceptable salts thereof, their use in pharmaceuticals, and compositions comprising the same.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to novel pyrido[4,3-d]pyrimidine compounds as inhibitors of Kirsten rat sarcoma virus oncogene homolog (KRAS). The present invention also relates to the preparation of the compounds and intermediates used in their preparation, compositions containing the compounds, and the use of the compounds for treating KRAS-related diseases such as cancer.

Background Art

[0002] KRAS, HRAS (Harvey rat sarcoma virus), and NRAS (neuroblastoma RAS viral oncogene homolog) belong to a group of GTPases that are very important in cell survival and proliferation through complex signaling cascades. Mutated RAS genes are found in approximately 30% of all cancers (Hyun et al., 2021 Int. J. Mol. Sci. 22(22), 12142). KRAS is the most frequently mutated RAS isoform (up to 85%) in cancer cells, leading to the development of cancers including non-small cell lung cancer (NSCLC), colorectal, and pancreatic cancer, which have a significant unmet medical need for both the population and the individual patients. KRAS mutations are widely seen in pancreatic ductal adenocarcinoma (PDAC). Mutations in KRAS are observed in 30% of NSCLC cases, the main form (80%) of lung cancer. The KRAS mutations found in NSCLC include 39% G12C, 18 - 21% G12V, and 17 - 18% G12D. KRAS mutations occur in 35 - 45% of colorectal cancers, leading to drug resistance.

[0003] Inhibitors of KRAS have been sought for decades, and the most recent advance has been the approval in trials of sotorasib and subsequent KRAS G12C-targeted compounds (Palmer et al., 2021 NPJ Precision Oncology, 5, 98). Sotorasib specifically targets the mutation in KRAS by covalent modification of the mutant cysteine at position 12. As such, sotorasib and other currently known KRAS inhibitors that rely on the same mechanism of action may have a narrow therapeutic range and limited use considering other major KRAS mutations such as G12V and G12D.

Summary of the Invention

Problems to be Solved by the Invention

[0004] Therefore, there remains a need for new KRAS inhibitors that can be used to treat a wider range of cancers.

Means for Solving the Problems

[0005] The present invention in part provides compounds of formula (I) to formula (VII), and pharmaceutically acceptable salts thereof. The compounds of the present invention can inhibit all activities of KRAS G12C, KRAS G12D, and KRAS G12V receptors, and can be useful in the treatment, prevention, suppression, and remission of diseases such as cancers, disorders, and pathologies mediated by any of KRAS G12C, KRAS G12D, and KRAS G12V receptors, or combinations thereof. The present invention also provides pharmaceutical compositions containing the compounds or salts of the present invention alone or in combination with additional anti-cancer therapeutic agents. The present invention also in part provides methods for preparing such compounds, pharmaceutically acceptable salts, and compositions of the present invention, as well as methods of using the foregoing. This summary is provided to introduce in a simplified form a selected number of the concepts that are further described below in the detailed description. This summary is not intended to identify key features or essential features of the subject matter recited in the claims, nor is it intended to be used in isolation as an aid in determining the scope of the subject matter recited in the claims.

[0006] According to one embodiment of the present invention, a compound of formula (I)

[0007]

Chemical formula

[0008]

Chemical formula

[0009] [Chemical formula] When it is as such, the piperazinyl ring is substituted with 1, 2 or 3 substituents independently selected from the group consisting of -OH, -CH2OH, -CN, -CH2CN, halogen, C1-C3 alkyl, C1-C3 fluoroalkyl, and C1-C3 alkoxy.

[0010] Embodiments of the present invention are described below. For convenience, Embodiment 1 (E1) is the same as the embodiment of formula (I) presented above.

[0011] It should be understood that both the above general description and the following detailed description are exemplary and explanatory only, and do not limit the present invention as described in the claims.

Brief Description of the Drawings

[0012]

Figure 1

Modes for Carrying Out the Invention

[0013] The present invention can be more easily understood by referring to the following detailed description of embodiments and examples of the present invention included in this specification. It should be understood that the present invention is not limited to the specific synthesis methods to be prepared, and of course the synthesis methods can vary. It should also be understood that the technical terms used in this specification are only for the purpose of describing specific embodiments and are not intended to be limiting.

[0014] E1 A compound of formula (I) as defined above or a pharmaceutically acceptable salt thereof.

[0015] E2 A compound of formula (I-a)

[0016]

Chemical formula

[0017] E3 A compound of embodiment E1 or embodiment E2, or a pharmaceutically acceptable salt thereof, wherein L is -O-CH2-.

[0018] E4 R 1 is 5- to 8-membered heterocycloalkyl containing 1 N as a single heteroatom, and 5- to 8-membered heterocycloalkyl may be substituted with 1, 2 or 3 substituents independently selected from the group consisting of halogen and C1-C3 alkyl, any one compound of embodiments E1 to E3, or a pharmaceutically acceptable salt thereof.

[0019] E5 R 1is

[0020] [Chem.] A compound of any one of Embodiments E1 to E4, or a pharmaceutically acceptable salt thereof, selected from the group consisting of

[0021] E6 R 1 is

[0022] [Chem.] A compound of Embodiment E5, or a pharmaceutically acceptable salt thereof, selected from the group consisting of

[0023] E7 R 2 is

[0024] [Chem.] [Wherein X and Y are each independently O or -CH2-, and R 2 is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of -OH and -CN] A compound of any one of Embodiments E1 to E6, or a pharmaceutically acceptable salt thereof, selected from the group consisting of

[0025] E8 R 2 is

[0026] [Chem.] A compound of Embodiment E7, or a pharmaceutically acceptable salt thereof, selected from the group consisting of

[0027] E9 R 2 is

[0028] [Chem.] A compound of Embodiment E8, or a pharmaceutically acceptable salt thereof, selected from the group consisting of

[0029] E10 R 2 is

[0030]

Chem.

[0031] E11 R 2 is

[0032]

Chem.

[0033] E12 R 2 is

[0034]

Chem.

[0035] E13 R 3 is C6 - C 10 a bicyclic aryl or a 4 - to 12 - membered bicyclic heteroaryl, and R 3 is a compound of any one of Embodiments E1 to E12, or a pharmaceutically acceptable salt thereof, which may be substituted with 1, 2, 3, or 4 substituents independently selected from the group consisting of -OH, halogen, -CN, C1 - C3 alkyl, C1 - C3 fluoroalkyl, and C2 - C3 alkynyl.

[0036] E14 R 3 is naphthyl which may be substituted with 1, 2, 3, or 4 substituents independently selected from the group consisting of -OH, halogen, -CN, C1 - C3 alkyl, C1 - C3 fluoroalkyl, and C2 - C3 alkynyl, a compound of Embodiment E13, or a pharmaceutically acceptable salt thereof.

[0037] E15 R 3 is

[0038]

Chemical formula

[0039] E16 R 4 is Cl or F, a compound of any one of Embodiments E1 to E15, or a pharmaceutically acceptable salt thereof.

[0040] E17

[0041]

Chemical formula

[0042]

Chemical formula

[0043] E18

[0044]

Chem.

[0045]

Chem.

[0046] E19 A pharmaceutical composition comprising a compound of any one of embodiments E1 to E18, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable additive.

[0047] E20 A method for treating cancer comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any one of embodiments E1 to E18, or a pharmaceutically acceptable salt thereof.

[0048] E21 A method for treating cancer comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any one of embodiments E1 to E18, or a pharmaceutically acceptable salt thereof, as a single agent.

[0049] E22 A method for treating cancer comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any one of embodiments E1 to E18, or a pharmaceutically acceptable salt thereof, and further comprising administering a therapeutically effective amount of an additional anti-cancer therapeutic agent.

[0050] E23 A method for treating cancer of any one of embodiments E20 to E22, wherein the cancer is small cell lung cancer (NSCLC), pancreatic cancer, or colorectal cancer.

[0051] E24 A compound of any one of embodiments E1 to E18, or a pharmaceutically acceptable salt thereof, for use as a medicament.

[0052] E25 A compound of any one of embodiments E1 to E18, or a pharmaceutically acceptable salt thereof, for use in the treatment of cancer.

[0053] E26 A compound for use in the treatment of cancer according to embodiment E25, wherein the cancer is small cell lung cancer (NSCLC), pancreatic cancer, or colorectal cancer.

[0054] E27 Use of a compound of any one of embodiments E1 to E18, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating cancer.

[0055] E28 Use of a compound, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating cancer according to embodiment E27, wherein the cancer is small cell lung cancer (NSCLC), pancreatic cancer, or colorectal cancer.

[0056] E29 A method for treating a disorder mediated by inhibition of the KRAS G12C, KRAS G12D, and KRAS G12V receptors in a subject, the method comprising administering to a subject in need thereof a compound of any one of embodiments E1 to E18, or a pharmaceutically acceptable salt thereof, in an amount effective to treat the disorder.

[0057] E30 A pharmaceutical combination comprising a compound of any one of embodiments E1 to E18, or a pharmaceutically acceptable salt thereof, and at least one additional therapeutic agent or a pharmaceutically acceptable salt thereof.

[0058] E31 A pharmaceutical composition comprising the pharmaceutical combination of Embodiment E30 and at least one additive.

[0059] E32 The compound of formula (II):

[0060] [Chemical formula] or a pharmaceutically acceptable salt thereof [wherein, R 1 is C3-C optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of -OH, -CN, halogen, C1-C3 alkyl, C1-C3 fluoroalkyl, and C1-C3 alkoxy 10 cycloalkyl or 4- to 12-membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from the group consisting of N, O, and S, R 2 is H or -(C1-C6 alkylene)-OH, -(C1-C6 alkylene)-CN, -(C1-C6 alkylene)-SH, -(C1-C3 alkylene)-S-(C1-C3 alkyl), -(C1-C3 alkylene)-(S=O)-(C1-C3 alkyl), -(C1-C3 alkylene)-(SO2)-(C1-C3 alkyl), C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 fluoroalkyl, C3-C6 fluorocycloalkyl, and C1-C6 alkoxy optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of -OH, -CN, -NH2, -SH, -(C1-C4 alkylene)-CN, -(C1-C4 alkylene)-OH, halogen, C1-C3 alkyl, C3-C6 cycloalkyl, C1-C3 fluoroalkyl, and C1-C3 alkoxy, and is selected from the group consisting of R 3 is C6-C optionally substituted with 1, 2, 3 or 4 substituents independently selected from the group consisting of -OH, halogen, -CN, C1-C3 alkyl, C1-C3 fluoroalkyl, C1-C3 alkoxy, and C2-C3 alkynyl 10Aryl or a 4- to 12-membered heteroaryl containing 1, 2, 3, or 4 N atoms, and R 3 When is substituted with two C1-C3 alkyl groups, the two C1-C3 alkyl groups may together form a 3- to 6-membered ring fused to the C6-C 10 aryl or 4- to 12-membered heteroaryl, R 4 is H, halogen, C1-C3 alkyl, or C1-C3 fluoroalkyl, R 5 is H, -OH, halogen, -NH2, CN, or is substituted with 1, 2, or 3 substituents independently selected from the group consisting of -OH, -CN, -NH2, -SH, -(C1-C4 alkylene)-CN, halogen, C1-C3 alkyl, C3-C6 cycloalkyl, C1-C3 fluoroalkyl, and C1-C3 alkoxy, and may be -(C1-C6 alkylene)-OH, -(C1-C6 alkylene)-CN, -(C1-C6 alkylene)-SH, -(C1-C3 alkylene)-S-(C1-C3 alkyl), -(C1-C3 alkylene)-(S=O)-(C1-C3 alkyl), -(C1-C3 alkylene)-(SO2)-(C1-C3 alkyl), C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 fluoroalkyl, C3-C6 fluorocycloalkyl, and C1-C6 alkoxy, or alternatively, R 5 and R 5 the carbon to which are attached, and R 2 and R 2 the nitrogen to which are attached together form a 4- to 8-membered heterocycloalkyl containing 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S, or a heteroatom-containing group selected from the group consisting of N(C1-C6 alkyl), -(S=O)-, and -(SO2)-, and the 4- to 8-membered heterocycloalkyl may be substituted with 1, 2, or 3 substituents selected from the group consisting of -OH, -CN, halogen, C1-C3 alkyl, -(C1-C6 alkylene)-CN, and -(C1-C6 alkylene)-OH, R 6is, each time it appears independently, H, -OH, halogen, CN, or is substituted with 1, 2 or 3 substituents independently selected from the group consisting of -OH, -CN, -(C1-C4 alkylene)-CN, halogen, C1-C3 alkyl, C3-C6 cycloalkyl, C1-C3 fluoroalkyl, and C1-C3 alkoxy, and is selected from the group consisting of -(C1-C6 alkylene)-OH, -(C1-C6 alkylene)-CN, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 fluoroalkyl, C3-C6 fluorocycloalkyl, and C1-C6 alkoxy; L is a linker containing 1, 2 or 3 members independently selected from the group consisting of -O-, -S-, -NR 7 -, and -CR 8 R 9 -; R 7 、R 8 、and R 9 are each independently H or C1-C3 alkyl; X is O, N, or S; l is 1 or 2; x is 1 or 2.

[0061] E33 A compound of embodiment E32, or a pharmaceutically acceptable salt thereof, wherein the linker L is -(O-CH2)-.

[0062] E34 R 1 is a 5- to 8-membered heterocycloalkyl containing 1 N as a single heteroatom, and the 5- to 8-membered heterocycloalkyl may be substituted with 1, 2 or 3 substituents independently selected from the group consisting of halogen and C1-C3 alkyl, a compound of embodiment E32 or embodiment E33, or a pharmaceutically acceptable salt thereof.

[0063] E35 R 1 is

[0064]

Chemical formula

[0065] E36 R 1 is

[0066]

Chemical formula

[0067] E37 R 2 is H, or is optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of -OH, -CN, and halogen, and is selected from the group consisting of -(C1-C5 alkylene)-OH and C1-C5 alkyl, a compound of any one of embodiments E32 to E36, or a pharmaceutically acceptable salt thereof.

[0068] E38 R 5 and R 6 are each independently selected from H, -OH, -CN, halogen, or are optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of -OH, -CN, and halogen, and are selected from the group consisting of -(C1-C5 alkylene)-OH, and a compound of any one of embodiments E32 to E37, or a pharmaceutically acceptable salt thereof.

[0069] E39 Formula (III):

[0070]

Chemical formula

[0071] E40 A compound of Embodiment E39, wherein Y is -CH2- or O.

[0072] E41 R 3 is a bicyclic aryl or bicyclic heteroaryl optionally substituted with 1 to 4 substituents each independently selected from the group consisting of -OH, halogen, -CN, C1-C3 alkyl, C1-C3 fluoroalkyl, and C2-C3 alkynyl, and when R 3 is substituted with two C1-C3 alkyl groups, the two C1-C3 alkyl groups may together form a 3- to 6-membered ring. A compound of any one of Embodiments E32 to E40, or a pharmaceutically acceptable salt thereof.

[0073] E42 R 3 is naphthyl optionally substituted with 1 to 4 substituents each independently selected from the group consisting of -OH, halogen, -CN, C1-C3 alkyl, C1-C3 fluoroalkyl, and C2-C3 alkynyl. A compound of Embodiment E41, or a pharmaceutically acceptable salt thereof.

[0074] E43 R 3 is

[0075]

Chemical Structure

[0076] E44 R 4 is a compound of any one of Embodiments E32 to E43, or a pharmaceutically acceptable salt thereof, wherein R is Cl or F.

[0077] E45 Formula (IV):

[0078] [Chemical formula] [wherein, R 3 is naphthyl which may be substituted with 1 to 4 substituents independently selected from the group consisting of -OH, halogen, -CN, C1-C3 alkyl, C1-C3 fluoroalkyl, and C2-C3 alkynyl, and l is 1 or 2] A compound of Embodiment E32 having the following formula, or a pharmaceutically acceptable salt thereof.

[0079] E46

[0080] [Chemical formula] A compound of Embodiment E32 selected from the group consisting of the following formula, or a pharmaceutically acceptable salt thereof.

[0081] E47

[0082] [Chemical formula] Or a compound which is a pharmaceutically acceptable salt thereof.

[0083] E48

[0084] [Chemical formula] Is a compound of the following formula.

[0085] E49

[0086] [Chemical] A pharmaceutically acceptable salt of a compound.

[0087] E50 A pharmaceutical composition comprising a compound of any one of Embodiments E32 to E47, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable additive.

[0088] E51 A method for treating cancer, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any one of Embodiments E32 to E47, or a pharmaceutically acceptable salt thereof.

[0089] E52 A method for treating cancer, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any one of Embodiments E32 to E47, or a pharmaceutically acceptable salt thereof as a single agent.

[0090] E53 A method for treating cancer, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any one of Embodiments E32 to E47, or a pharmaceutically acceptable salt thereof, and further comprising administering a therapeutically effective amount of an additional anti-cancer therapeutic agent.

[0091] E54 A method for treating cancer according to any one of Embodiments E51 to E53, wherein the cancer is non-small cell lung cancer (NSCLC), pancreatic cancer, or colorectal cancer.

[0092] E55 A compound of any one of Embodiments E32 to E47, or a pharmaceutically acceptable salt thereof, for use as a medicament.

[0093] E56 A compound of any one of Embodiments E32 to E47, or a pharmaceutically acceptable salt thereof, for use in the treatment of cancer.

[0094] E57 A compound for use in the treatment of cancer according to embodiment E56, wherein the cancer is non-small cell lung cancer (NSCLC), pancreatic cancer, or colorectal cancer.

[0095] E58 Use of a compound of any one of embodiments E32 to E47, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating cancer.

[0096] E59 Use of a compound, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating cancer according to embodiment E58, wherein the cancer is non-small cell lung cancer (NSCLC), pancreatic cancer, or colorectal cancer.

[0097] E60 A method for treating a disorder mediated by inhibition of the KRAS G12C, KRAS G12D, and KRAS G12V receptors in a subject, the method comprising administering to a subject in need thereof a compound of any one of embodiments E32 to E47, or a pharmaceutically acceptable salt thereof, in an amount effective to treat the disorder.

[0098] E61 A pharmaceutical combination comprising a compound of any one of embodiments E32 to E47, or a pharmaceutically acceptable salt thereof, and at least one additional therapeutic agent or a pharmaceutically acceptable salt thereof, the combination being a fixed or non-fixed combination.

[0099] E62 A pharmaceutical composition comprising the pharmaceutical combination of embodiment E61 and at least one additive.

[0100] E63 A compound of formula (V):

[0101]

Chemical formula

[0102]

Chem.

[0103]

Chem.

[0104] E64 R 1 is

[0105] [Chem.] a compound of Embodiment E63 selected from the group consisting of, or a pharmaceutically acceptable salt thereof.

[0106] E65 R 1 is

[0107] [Chem.] a compound of Embodiment E64, or a pharmaceutically acceptable salt thereof.

[0108] E66 R 2 is C3 alkyl, and R 5 is -(C1 alkylene)-OH, a compound of any one of Embodiments E63 to E65, or a pharmaceutically acceptable salt thereof.

[0109] E67 R 2 is -(C3 alkylene)-OH, and R 5 is C1 alkyl, a compound of any one of Embodiments E63 to E65, or a pharmaceutically acceptable salt thereof.

[0110] E68 R 2 and R 5 together form a 7-membered heterocycloalkyl containing one heteroatom O, a compound of Embodiment E66, or a pharmaceutically acceptable salt thereof.

[0111] E69 R 2 and R 5 together form a 7-membered heterocycloalkyl containing one heteroatom O, a compound of Embodiment E67, or a pharmaceutically acceptable salt thereof.

[0112] E70 R 3 is

[0113]

Chem.

[0114] E71 R 4 is F, a compound of any one of Embodiments E63 to E70, or a pharmaceutically acceptable salt thereof.

[0115] E72 X is O and l is 1, a compound of any one of Embodiments E63 to E71, or a pharmaceutically acceptable salt thereof.

[0116] E73 Formula (VI):

[0117]

Chem.

[0118] E74 R 2 is C3 alkyl, and R 5 is -(C1 alkylene)-OH, a compound of Embodiment E73, or a pharmaceutically acceptable salt thereof.

[0119] E75 R 2 is -(C3 alkylene)-OH, and R 5 is C1 alkyl, a compound of Embodiment E73, or a pharmaceutically acceptable salt thereof.

[0120] E76 R 2 and R 5The compound of embodiment E74, or a pharmaceutically acceptable salt thereof, in which they combine to form a 7-membered heterocycloalkyl containing one heteroatom O.

[0121] E77 R 2 and R 5 The compound of embodiment E75, or a pharmaceutically acceptable salt thereof, in which they combine to form a 7-membered heterocycloalkyl containing one heteroatom O.

[0122] E78 The compound of formula (VII)

[0123]

Chemical formula

[0124] E79 R 11 , R 12 , R 13 , and R 14 The compound of embodiment E78, or a pharmaceutically acceptable salt thereof, wherein they are each independently H or methyl.

[0125] E80

[0126]

Chemical formula

[0127] E81

[0128]

Chemical formula

[0129] E82 A pharmaceutical composition comprising a compound of any one of Embodiments E63 to E81, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable additive.

[0130] E83 A method for treating cancer, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any one of Embodiments E63 to E81, or a pharmaceutically acceptable salt thereof.

[0131] E84 A method for treating cancer, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any one of Embodiments E63 to E81, or a pharmaceutically acceptable salt thereof, as a single agent.

[0132] E85 A method for treating cancer, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any one of Embodiments E63 to E81, or a pharmaceutically acceptable salt thereof, and further comprising administering a therapeutically effective amount of an additional anti-cancer therapeutic agent.

[0133] E86 A method for treating any one of the cancers of Embodiments E83 to E85, wherein the cancer is small cell lung cancer (NSCLC), pancreatic cancer, or colorectal cancer.

[0134] E87 A compound of any one of Embodiments E63 to E81, or a pharmaceutically acceptable salt thereof, for use as a medicament.

[0135] E88 A compound of any one of Embodiments E63 to E81, or a pharmaceutically acceptable salt thereof, for use in the treatment of cancer.

[0136] E89 A compound for use in the treatment of cancer according to embodiment E88, wherein the cancer is non-small cell lung cancer (NSCLC), pancreatic cancer, or colorectal cancer.

[0137] E90 Use of a compound according to any one of embodiments E63 to E81, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating cancer.

[0138] E91 Use of a compound, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating cancer according to embodiment E90, wherein the cancer is non-small cell lung cancer (NSCLC), pancreatic cancer, or colorectal cancer.

[0139] E92 A method for treating a disorder mediated by inhibition of the KRAS G12C, KRAS G12D, and KRAS G12V receptors in a subject, the method comprising administering to a subject in need thereof a compound according to any one of embodiments E63 to E81, or a pharmaceutically acceptable salt thereof, in an amount effective to treat the disorder.

[0140] E93 A pharmaceutical combination comprising a compound according to any one of embodiments E63 to E81, or a pharmaceutically acceptable salt thereof, and at least one additional therapeutic agent or a pharmaceutically acceptable salt thereof, the combination being a fixed or non-fixed combination.

[0141] E94 A pharmaceutical composition comprising the pharmaceutical combination of embodiment E93 and at least one additive.

[0142] Each of the embodiments described herein can be combined with any other embodiment that is not inconsistent with the combined embodiments described herein. Additionally, any of the compounds described in the examples, or a pharmaceutically acceptable salt thereof, may be claimed individually or as a group together with one or more other compounds of the examples, or a pharmaceutically acceptable salt thereof.

[0143] Furthermore, each of the embodiments described herein contemplates, within its scope, pharmaceutically acceptable salts of the compounds described herein, stereoisomers of the compounds, and pharmaceutically acceptable salts of the stereoisomers.

[0144] Definitions Unless defined otherwise herein, scientific and technical terms used in connection with the present invention have the meanings commonly understood by one of ordinary skill in the art.

[0145] The invention described herein can be practiced appropriately even in the absence of any element not specifically disclosed herein.

[0146] "Compounds of the invention" include compounds of formula (I) and novel intermediates used in their preparation. One of ordinary skill in the art will recognize that the compounds of the invention include, where possible, conformational isomers (e.g., cis and trans isomers) and all optical isomers (e.g., enantiomers and diastereoisomers), racemates, diastereoisomers and other mixtures of such isomers, and tautomers thereof. One of ordinary skill in the art will also recognize that the compounds of the invention include, where formed, solvates, hydrates, polymorphs, pseudopolymorphs, esters, salt forms, prodrugs, and isotopically labeled versions thereof.

[0147] As used herein, the singular forms "a", "an" and "the" include plural references unless the context clearly dictates otherwise. For example, "a" substituent includes one or more substituents.

[0148] As used herein, the term "about", when used to modify a parameter defined by a numerical value (e.g., a dose of 5 mg), means that the parameter can vary up to 10% above or below the numerical value described for that parameter. For example, a dose of about 5 mg means 5 mg ± 10%, i.e., it may vary between 4.5 mg and 5.5 mg.

[0149] When a substituent is described as being "independently selected" from a group, each substituent is selected independently of the others. Thus, each substituent may be the same as or different from the other substituents.

[0150] "Optional" or "optionally" means that the event or circumstance described later may occur, but need not occur, and that the description includes cases where the event or circumstance occurs and cases where it does not occur.

[0151] The terms "optionally substituted" and "substituted or unsubstituted" are used interchangeably to indicate that the particular group described may or may not have a non-hydrogen substituent (i.e., be unsubstituted), or that the group may have one or more non-hydrogen substituents (i.e., be substituted). Unless otherwise specified, the total number of substituents that may be present is equal to the number of H atoms present on the unsubstituted form of the group described. When an optional substituent, such as an oxo (=O) substituent, is attached via a double bond, that group occupies two available valences, and thus the total number of other substituents included is reduced by two. When optional substituents are independently selected from a list of options, the groups selected may be the same or different. Throughout the present disclosure, it will be understood that the number and nature of optional substituents will be limited only insofar as such substitution makes chemical sense to one of ordinary skill in the art.

[0152] "Halogen" refers to fluoro, chloro, bromo and iodo (F, Cl, Br, I).

[0153] "Cyano" refers to a substituent having a carbon atom that is part of a triple bond with a nitrogen atom, i.e., -C≡N (also denoted herein as "-CN").

[0154] "Hydroxy" refers to the -OH group.

[0155] "Oxo" refers to a double-bonded oxygen (=O).

[0156] "Alkyl" refers to a saturated monovalent aliphatic hydrocarbon radical having the specified number of carbon atoms, including straight-chain or branched-chain groups. Alkyl groups may contain, but are not limited to, 1 to 6 carbon atoms ("C1-C6 alkyl"), 1 to 3 carbon atoms ("C1-C3 alkyl"), or 1 to 2 carbon atoms ("C1-C2 alkyl"). Examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, and the like.

[0157] "Fluoroalkyl" refers to an alkyl group as defined herein, in which one or all of the hydrogen atoms of the alkyl group are replaced by fluorine atoms. Examples include, but are not limited to, fluoromethyl, difluoromethyl, fluoroethyl, difluoroethyl, trifluoroethyl, and tetrafluoroethyl. Examples of fully substituted fluoroalkyl groups (also referred to as perfluoroalkyl groups) include trifluoromethyl (-CF3) and pentafluoroethyl (-C2F5).

[0158] "Alkylene" refers to a divalent aliphatic hydrocarbon radical having the specified number of carbon atoms. Alkylene groups may contain, but are not limited to, 1 to 6 carbon atoms ("C1-C6 alkylene"), or 1 to 2 carbon atoms ("C1-C2 alkylene"). Examples include -(CH2)- (methylene) and -(CH2-CH2)- (ethylene).

[0159] "Alkoxy" refers to an alkyl group as defined herein that is singly bonded to an oxygen atom. The point of attachment of the alkoxy radical to the molecule is via the oxygen atom. An alkoxy radical may also be denoted as alkyl-O-. The alkoxy group may contain, but is not limited to, 1 to 6 carbon atoms ("C1-C6 alkoxy"), or 1 to 3 carbon atoms ("C1-C3 alkoxy"). The alkoxy group includes, but is not limited to, methoxy, ethoxy, n-propoxy, and the like.

[0160] "Alkynyl" refers to an alkyl group as defined herein that consists of at least 2 carbon atoms and at least 1 carbon-carbon triple bond. Alkynyl may contain 2 to 3 carbon atoms ("C2-C3 alkynyl"). Examples include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, and the like.

[0161] "Cycloalkyl" refers to a completely saturated hydrocarbon ring system that may be a monocyclic bridged or fused bicyclic or polycyclic ring system having the specified number of carbon atoms and is linked to the base molecule through the carbon atoms of the cycloalkyl ring. The cycloalkyl group may contain, but is not limited to, 3 to 10 carbon atoms ("C3-C 10 Cycloalkyl"), 3 to 8 carbon atoms ("C3-C8 cycloalkyl"), 3 to 6 carbon atoms ("C3-C6 cycloalkyl"), 3 to 5 carbon atoms ("C3-C5 cycloalkyl") or 3 to 4 carbon atoms ("C3-C4 cycloalkyl"). Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantanyl, and the like. The cycloalkyl group may be substituted, unsubstituted, or may be substituted as further defined herein.

[0162] "Fluorocycloalkyl" refers to a cycloalkyl group as defined herein in which one to all of the hydrogen atoms of the alkyl group are replaced by fluorine atoms. Examples include, but are not limited to, fluorocyclopropyl, fluorocyclobutyl, fluorocyclopentyl, and fluorocyclohexyl.

[0163] "Heterocycloalkyl" refers to a fully saturated ring system containing a specified number of ring atoms, containing at least one heteroatom selected from N, O, and S as ring members, where the ring S atom may be substituted by one or two oxo groups (i.e., S(O) q , where q is 0, 1, or 2), and the heterocycloalkyl ring is linked to the base molecule through a ring atom which may be C or N. The heterocycloalkyl ring includes monocyclic or polycyclic rings such as bicyclic rings. The heterocycloalkyl ring may be spirocyclic, bridged, or fused to one or more other heterocycloalkyl or carbocyclic rings, and such spirocyclic, bridged, or fused rings themselves may also be saturated, unsaturated, or partially unsaturated to the extent that the chemical meaning of aromaticity is achieved, or aromatic, provided that the point of attachment to the base molecule is an atom of the heterocycloalkyl portion of the ring system. The heterocycloalkyl ring may contain 1 to 4 heteroatoms selected from N, O, and S(O) q , or 1 to 3 ring heteroatoms, or 1 to 2 ring heteroatoms, provided that such a heterocycloalkyl ring does not contain two adjacent oxygen or sulfur atoms.

[0164] The heterocycloalkyl ring may be further substituted as defined herein, may be unsubstituted, or may be substituted. Such substituents may be present on or attached to the heterocyclic ring attached to the base molecule, or on a monocyclic, bicyclic, tricyclic, spirocyclic, bridged, or fused ring attached thereto.

[0165] The heterocycloalkyl ring may include, but is not limited to, a 4- to 12-membered heterocyclyl group, such as a 5- to 8- or 4- to 6-membered heterocycloalkyl group, according to the definitions herein. Examples of the heterocycloalkyl ring group of the present invention may include, but are not limited to, aziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, azepanyl, oxaazepanyl, thieazepanyl, radicals of a hexahydro-1H-pyrrolidine ring, radicals of an 8-oxa-3-azabicyclo[3.2.1]octane ring, radicals of a 3-azabicyclo[3.2.1]octane ring, radicals of a 6-azabicyclo[3.2.1]octane ring, or radicals of a 3-azabicyclo[3.2.0]heptane ring.

[0166] "Aryl" or "aromatic" contains a specified number of ring atoms, and all carbon atoms in the ring are sp 2 Hybridized, and the pi electrons are conjugated, referring to a monocyclic, bicyclic (e.g., biaryl, fused) or polycyclic ring system. The aryl group may contain, but is not limited to, 6 to 10 carbon atoms ("C6~C 10 Aryl"). The fused aryl group may include an aryl ring (e.g., a phenyl ring) fused to another aryl ring. Examples include, but are not limited to, phenyl, biphenyl, naphthyl, anthracenyl, phenanthrenyl, indanyl, and indenyl. The aryl group may be substituted, unsubstituted, or optionally substituted as further defined herein.

[0167] Similarly, "heteroaryl" or "heteroaromatic" contains a specified number of ring atoms, includes at least one heteroatom selected from N, O, and S as a ring member in the ring, and all carbon atoms in the ring are sp 2Refers to a monocyclic, bicyclic (e.g., heteroaryl, fused), or polycyclic ring system that is hybridized and has conjugated π electrons. The heteroaryl group may contain, but is not limited to, 5 to 14 ring atoms (“5- to 14-membered heteroaryl”), 5 to 12 ring atoms (“5- to 12-membered heteroaryl”), 5 to 10 ring atoms (“5- to 10-membered heteroaryl”), 5 to 9 ring atoms (“5- to 9-membered heteroaryl”), or 5 to 6 ring atoms (“5- to 6-membered heteroaryl”). The heteroaryl ring is attached to the base molecule through the ring atoms of the heteroaromatic ring. Thus, either a 5- or 6-membered heteroaryl ring may be attached to the base molecule alone or in a fused structure through a ring C or N atom.Examples of heteroaryl groups include, but are not limited to, pyrrolyl, furanyl, thiophenyl, pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, isothiazolyl, thiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, pyridinyl, pyridizinyl, pyrimidinyl, pyrazinyl, benzofuranyl, benzothiophenyl, indolyl, benzimidazolyl, indazolyl, benzotriazolyl, pyrrolo[2,3-b]pyridinyl, pyrrolo[2,3-c]pyridinyl, pyrrolo[3,2-c]pyridinyl, pyrrolo[3,2-b]pyridinyl, quinolinyl, isoquinolinyl, purinyl, triazinyl, naphthyridinyl, cinnolinyl, quinazolinyl, quinoxalinyl, carbazolyl, imidazo[4,5-b]pyridinyl, imidazo[4,5-c]pyridinyl, pyrazolo[4,3-d]pyridinyl, pyrazolo[4,3-c]pyridinyl, pyrazolo[3,4-c]pyridinyl, pyrazolo[3,4-b]pyridinyl, isoindolyl, purinyl, indolinyl, imidazo[1,2-a]pyridinyl, imidazo[1,5-a]pyridinyl, pyrazolo[1,5-a]pyridinyl, pyrrolo[1,2-b]pyridazinyl, imidazo[1,2-c]pyrimidinyl, azaquinazolinyl, phthalazinyl, (pyrido[3,2-d]pyrimidinyl, (pyrido[4,3-d]pyrimidinyl, (pyrido[3,4-d]pyrimidinyl, (pyrido[2,3-d]pyrimidinyl, pyrido[2,3-b]pyrazinyl, pyrido[3,4-b]pyrazinyl, pyrimido[5,4-d]pyrimidinyl, pyrazino[2,3-b]pyrazinyl, pyrimido[4,5-d]pyrimidinyl. Examples of 5- or 6-membered heteroaryl groups include, but are not limited to, pyrrolyl, furanyl, thiophenyl, pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, isothiazolyl, thiazolyl, triazolyl, pyridinyl, pyrimidinyl, pyrazinyl and pyridazinyl rings. The heteroaryl group may be optionally substituted, unsubstituted or may be substituted as further defined herein.

[0168] "Amino" refers to an unsubstituted -NH2 group. When amino is described as substituted or optionally substituted, the term includes groups of the form -NRxRy, where each of Rx and Ry is defined as further described herein. For example, "alkylamino" refers to an -NRxRy group in which one of Rx and Ry is an alkyl moiety and the other is H, and "dialkylamino" refers to an -NRxRy in which both Rx and Ry are alkyl moieties and the alkyl moiety has the specified number of carbon atoms (e.g., -NH(C1-C4 alkyl) or -N(C1-C4 alkyl)2).

[0169] In the present disclosure, the wavy line "

[0170]

Chemical formula

[0171] The term "pharmaceutically acceptable" means that a substance (e.g., a compound described herein) and any salt thereof, or a composition containing the substance or salt of the present invention, is suitable for administration to a subject or patient.

[0172] As used herein, the "deuterium enrichment factor" means the ratio between the deuterium abundance and the natural abundance of deuterium relative to the hydrogen abundance, respectively. The atomic arrangements shown to have deuterium typically have a deuterium enrichment factor of at least 1000 (15% deuterium introduced), at least 2000 (30% deuterium introduced), at least 3000 (45% deuterium introduced), at least 3500 (52.5% deuterium introduced), at least 3500 (52.5% deuterium introduced with each indicated deuterium atom), at least 4000 (60% deuterium introduced), at least 4500 (67.5% deuterium introduced), at least 5000 (75% deuterium introduced), at least 5500 (82.5% deuterium introduced), at least 6000 (90% deuterium introduced), at least 6333.3 (95% deuterium introduced), at least 6466.7 (97% deuterium introduced), at least 6600 (99% deuterium introduced), or at least 6633.3 (99.5% deuterium introduced) in certain embodiments.

[0173] Salt Salts included within the scope of the term "pharmaceutically acceptable salts" generally refer to compounds of the present invention prepared by reacting a free base or a free acid, respectively, with a suitable organic or inorganic acid, or a suitable organic or inorganic base, to obtain a salt of the compound of the present invention that is suitable for administration to a subject or patient.

[0174] In addition, the compounds of formula (I) may also include other salts of such compounds, which are not necessarily pharmaceutically acceptable salts and may be useful as intermediates for one or more of the following: 1) for preparing the compounds of formula (I); 2) for purifying the compounds of formula (I); 3) for separating the enantiomers of the compounds of formula (I); or 4) for separating the diastereoisomers of the compounds of formula (I).

[0175] Suitable acid addition salts are formed from acids that form non-toxic salts. Examples include, but are not limited to, acetate, adipate, aspartate, benzoate, besylate, bicarbonate / carbonate, bisulfate / sulfate, borate, camsylate, citrate, cyclamate, edisylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, maleate, malonate, mesylate, methylsulfate, naphthylate, 2-naphthylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogenphosphate / dihydrogenphosphate, pyroglutamate, saccharinate, stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate, 1,5-naphthalenedisulfonic acid, and xinoformate.

[0176] Suitable base salts are formed from bases that form non-toxic salts. Examples include, but are not limited to, aluminum, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine, and zinc salts.

[0177] Half salts of acids and bases, such as half sulfate salts and half calcium salts, can also be formed.

[0178] For an overview of suitable salts, see Paulekun, G.S. et al., Trends in Active Pharmaceutical Ingredient Salt Selection Based on Analysis of the Orange Book Database, J. Med. Chem. 2007;50(26), 6665 - 6672.

[0179] Pharmaceutically acceptable salts of the compounds of the present invention can be prepared by methods well known to those skilled in the art, including, but not limited to, the following procedures. (i) A procedure by reacting the compound of the present invention with a desired acid or base; (ii) A procedure by removing an acid- or base-labile protecting group from a suitable precursor of the compound of the present invention or by ring-opening a suitable cyclic precursor, such as a lactone or lactam, using a desired acid or base; or (iii) A procedure by converting one salt of the compound of the present invention into another salt. This can be achieved by reacting with a suitable acid or base or by using a suitable ion-exchange procedure.

[0180] These procedures are typically carried out in solution. The resulting salt can be precipitated and collected by filtration or recovered by evaporation of the solvent.

[0181] Solvate The compounds of the present invention, and their pharmaceutically acceptable salts, may exist in unsolvated and solvated forms. The term "solvate" is used herein to describe a molecular complex comprising a compound of the present invention, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable solvent molecules, such as ethanol. The term "hydrate" is used when the solvent is water.

[0182] In addition, the compounds of formula (I) may include other solvates of such compounds, which are not necessarily pharmaceutically acceptable solvates and may be useful as intermediates for one or more of the following: 1) for preparing the compounds of formula (I); 2) for purifying the compounds of formula (I); 3) for separating enantiomers of the compounds of formula (I); or 4) for separating diastereoisomers of the compounds of formula (I).

[0183] The currently recognized classification system for organic hydrates is a classification system that defines isolated sites, channels, or metal ion coordination hydrates. See Polymorphism in Pharmaceutical Solids by K.R. Morris (edited by H.G. Brittain, Marcel Dekker, 1995). Isolated site hydrates are hydrates in which water molecules are isolated from direct contact with each other by the intervention of organic molecules. In channel hydrates, water molecules are present within the lattice channels and are adjacent to other water molecules there. In metal ion coordination hydrates, water molecules are bonded to metal ions.

[0184] When the solvent or water is tightly bound, the complex can have a well-defined stoichiometry independent of humidity. However, when the binding of the solvent or water is weak, as in channel solvates and hygroscopic compounds, the water / solvent content can be influenced by humidity and the drying state. In such cases, non-stoichiometry becomes the norm.

[0185] Complex Multi-component complexes (other than salts and solvates) in which the drug and at least one other component are present in stoichiometric or non-stoichiometric amounts are also included within the scope of the present invention. This type of complex includes clathrates (drug-host inclusion complexes) and co-crystals. The latter are typically defined as crystalline complexes of neutral molecular components held together by non-covalent interactions. For example, hydrogen bond complexes (co-crystals) can be formed with neutral molecules or salts. Co-crystals can be prepared by melt crystallization, recrystallization from solvents, or physical grinding of the components. See Chem Commun, 17; 1889 - 1896 by O. Almarsson and M.J. Zaworotko (2004). For a general review of multi-component complexes, see J Pharm Sci, 64(8), 1269 - 1288 by Haleblian (August 1975).

[0186] Solid form The compounds of the present invention can exist in a continuous solid state ranging from completely amorphous to completely crystalline. The term "amorphous" refers to a state in which the material lacks long-range order at the molecular level and can exhibit physical properties of a solid or a liquid depending on temperature. Typically, such materials do not exhibit a characteristic X-ray diffraction pattern and, while showing solid properties, are more formally described as liquids. Upon heating, a change from solid to liquid properties occurs, which is typically characterized by a secondary state change ("glass transition"). The term "crystalline" refers to a solid phase in which the material has an internal structure with regularly arranged molecules at the molecular level and exhibits a characteristic X-ray diffraction pattern with defined peaks. When such materials are heated sufficiently, they also exhibit liquid properties, but the change from solid to liquid is typically characterized by a primary phase change ("melting point").

[0187] The compounds of the present invention can also exist in an intermediate state (mesophase or liquid crystal) when placed under suitable conditions. The intermediate state is intermediate between a true crystalline state and a true liquid state (either a melt or a solution) and consists of two-dimensional order at the molecular level. An intermediate state resulting from a temperature change is described as "thermotropic", and one resulting from the addition of a second component such as water or another solvent is described as "lyotropic". Compounds having the potential to form lyotropic mesophases are described as "amphiphilic" and consist of molecules with polar head groups such as ionic (-COO - Na + 、-COO - K + 、or -SO3 - Na + etc.) or non-ionic (-N - N + (CH3)3 etc.). For further information, see Crystals and the Polarizing Microscope, N.H. Hartshorne and A. Stuart, 4th edition (Edward Arnold, 1970).

[0188] Stereoisomers The compounds of the present invention may exist as two or more stereoisomers. The stereoisomers of the compounds may include cis and trans isomers (geometric isomers), optical isomers such as R and S enantiomers, diastereoisomers, rotational isomers, atropisomers, and conformational isomers. For example, the compounds of the present invention containing one or more asymmetric carbon atoms may exist as two or more stereoisomers.

[0189] The pharmaceutically acceptable salts of the compounds of the present invention may also contain counterions that are optically active (e.g., d-lactate or l-lysine) or racemic (e.g., dl-tartrate or dl-arginine).

[0190] Cis / trans isomers can be separated by conventional techniques well known to those skilled in the art, such as chromatography and fractional crystallization.

[0191] Conventional techniques for preparing / isolating individual enantiomers include chiral synthesis from a suitable optically pure precursor, or resolution of a racemate (or a racemate of a salt or derivative), for example, using chiral high performance liquid chromatography (HPLC). Alternatively, a racemate (or racemic precursor) can be reacted with a suitable optically active compound, such as an alcohol, or, if the compound of the invention contains an acidic or basic moiety, a base or acid such as 1-phenylethylamine or tartaric acid. The resulting mixture of diastereoisomers can be separated by chromatography, fractional crystallization, or by using both of the above techniques, and one or both of the diastereoisomers can be converted to the corresponding pure enantiomers by means well known to those skilled in the art. Chromatography can be used to obtain the chiral compounds of the invention (and their chiral precursors) in enantiomerically enriched form, typically by HPLC concentration of the eluent to give an enriched mixture. Chiral chromatography using subcritical and supercritical fluids can be used. Methods for chiral chromatography useful in some embodiments of the invention are known in the art (see, for example, Smith, Roger M., Loughborough University, Loughborough, UK; Chromatographic Science Series (1998), 75 (Supercritical Fluid Chromatography with Packed Columns), pages 223-249 and the references cited therein).

[0192] When any racemate crystallizes, two different types of crystals can occur. The first type is the above-mentioned racemic compound (true racemic compound) in which one uniform form of crystal containing both enantiomers in equimolar amounts is formed. The second type is a racemic mixture or conglomerate in which two crystal forms each containing a single enantiomer occur in equimolar amounts. Both of the crystal forms present in the racemic mixture have the same physical properties, but they may have different physical properties compared to the true racemate. Racemic mixtures can be separated by conventional techniques known to those skilled in the art. See, for example, Stereochemistry of Organic Compound by E. L. Eliel and S. H. Wilen (Wiley, New York, 1994).

[0193] Tautomerism Tautomeric isomerism ( "tautomerism") can occur when structural isomers are interconvertible via a low energy barrier. This can take the form of, for example, proton tautomerism in compounds of the invention containing imino / amino, keto / enol, or oxime / nitroso groups, lactam / lactim, or so-called valence tautomerism in compounds containing an aromatic moiety. It follows that a single compound can exhibit more than one type of isomerism.

[0194] For the sake of brevity, the compounds of the invention are illustrated herein in a single tautomeric form, but it is important to emphasize that all possible tautomeric forms are included within the scope of the invention.

[0195] Isotope The present invention includes all pharmaceutically acceptable isotopically labeled compounds of the invention in which one or more atoms are replaced by atoms having the same atomic number but a different atomic mass or mass number than the atomic mass or mass number that is predominant in nature.

[0196] Examples of isotopes suitable for inclusion in the compounds of the invention include2 H and 3 hydrogen such as 11 C, 13 C and 14 carbon such as 36 chlorine such as 18 fluorine such as 123 I and 125 iodine such as 13 N and 15 nitrogen such as 15 O, 17 O and 18 oxygen such as 32 phosphorus such as 35 sulfur isotopes may be included.

[0197] Certain isotope-labeled compounds of the present invention, for example, those incorporating a radioisotope, are useful in one or both of drug or substrate tissue distribution studies. Radioisotope tritium, i.e., 3 H, and carbon-14, i.e., 14 C are particularly useful for this purpose considering their ease of incorporation and rapid means of detection.

[0198] Deuterium, i.e., 2 substitution with H can provide certain therapeutic advantages resulting from greater metabolic stability.

[0199] Positron-emitting radioisotopes, for example 11 C, 18 F, 15 O and 13 substitution with N may be useful in positron emission tomography (PET) studies for examining substrate receptor occupancy.

[0200] In some embodiments, the present disclosure provides deuterium-labeled (or deuterated) compounds and salts, wherein the formulas and variables of such compounds and salts are each and independently as described herein. "Deuterated" means that at least one of the atoms in the compound is deuterium at an abundance higher than the natural abundance of deuterium (typically, approximately 0.015%). Skilled practitioners recognize that in chemical compounds containing hydrogen atoms, the hydrogen atoms actually exist as a mixture of H and D, with approximately 0.015% being D. The concentration of deuterium incorporated into the deuterium-labeled compounds and salts of the present invention can be defined by a deuterium enrichment factor. It is understood that one or more deuteriums can exchange with hydrogen under physiological conditions.

[0201] In some embodiments, the deuterium compound is selected from any one of the compounds described in Table 2 shown in the Examples section.

[0202] In some embodiments, one or more hydrogen atoms on certain metabolic sites on the compounds of the present invention are deuterated.

[0203] The isotopically labeled compounds of the present invention can generally be prepared by using appropriate isotopically labeled reagents in place of the previously used unlabeled reagents by conventional techniques known to those skilled in the art or by processes similar to those described in the attached Examples and Preparation Examples.

[0204] Pharmaceutically acceptable solvates according to the present invention include those in which the crystallization solvent may be isotopically substituted, for example, D2O, d6-acetone, d6-DMSO.

[0205] Prodrug The compounds of the present invention can be administered in the form of prodrugs. Thus, certain derivatives of the compounds of the present invention, which may themselves have little or no pharmacological activity, can be converted into the compounds of the present invention having the desired activity when administered in or on the body, for example, by hydrolytic cleavage, particularly hydrolytic cleavage facilitated by esterase or peptidase enzymes. Such derivatives are referred to as "prodrugs". Further information on the use of prodrugs can be found in "The Expanding Role of Prodrugs in Contemporary Drug Design and Development, Nature Reviews Drug Discovery, 17, 559-587 (2018) (J. Rautio et al.).

[0206] The prodrugs according to the present invention can be produced, for example, as described in "Design of Prodrugs" by H. Bundgaard (Elsevier, 1985), by replacing appropriate functional groups present in the compounds of the present invention with certain parts known to those skilled in the art as "pro parts".

[0207] Thus, the prodrugs according to the present invention can be (a) ester or amide derivatives of carboxylic acids when present in the compounds of the present invention; (b) ester, carbonate, carbamate, phosphate or ether derivatives of hydroxyl groups when present in the compounds of the present invention; (c) amide, imine, carbamate or amine derivatives of amino groups when present in the compounds of the present invention; (d) thioester, thiocarbonate, thiocarbamate or sulfide derivatives of thiol groups when present in the compounds of the present invention; or (e) oxime or imine derivatives of carbonyl groups when present in the compounds of the present invention.

[0208] Some specific examples of the prodrugs according to the present invention include: (i) When the compound of the present invention contains a carboxylic acid functional group (-COOH), its ester, for example, the hydrogen of the carboxylic acid functional group of the compound is replaced by C1-C8 alkyl (for example, ethyl) or (C1-C8 alkyl)C(=O)OCH2- (for example, t BuC(=O)OCH2-); (ii) When the compound of the present invention contains an alcohol functional group (-OH), its ester, for example, the hydrogen of the alcohol functional group of the compound is replaced by -CO(C1-C8 alkyl) (for example, methylcarbonyl), or the alcohol is esterified with an amino acid; (iii) When the compound of the present invention contains an alcohol functional group (-OH), its ether, for example, the hydrogen of the alcohol functional group of the compound is replaced by (C1-C8 alkyl)C(=O)OCH2- or -CH2OP(=O)(OH)2; (iv) When the compound of the present invention contains an alcohol functional group (-OH), its phosphate, for example, the hydrogen of the alcohol functional group of the compound is replaced by -P(=O)(OH)2 or -P(=O)(O - Na + )2 or -P(=O)(O - )2Ca 2+ ; (v) When the compound of the present invention contains a primary or secondary amino functional group (-NH2 or -NHR, where R≠H), its amide, for example, in some cases, one or both of the hydrogens of the amino functional group of the compound are replaced by (C1-C 10 ) alkanoyl, -COCH2NH2, or the amino group is derivatized with an amino acid; (vi) When the compound of the present invention contains a primary or secondary amino functional group (-NH2 or -NHR, where R≠H), its amine, for example, in some cases, one or both of the hydrogens of the amino functional group of the compound are replaced by -CH2OP(=O)(OH)2; (vii) When the compound of the present invention contains an alcohol functional group (-OH), replacement of the hydrogen of the alcohol functional group with a group selected from the following set:

[0209] [Chemical formula] [wherein, R, R’, R”, R”’ are (C1-C8) alkyl or (C1-C8) alkoxy, which may be linear, branched or cyclic] is included.

[0210] Some preferred prodrugs can be prepared via -OH on a C6-C 10 bicyclic aryl or 4- to 12-membered bicyclic heteroaryl. Some more preferred prodrugs can be prepared via -OH on naphthyl.

[0211] Certain compounds of the present invention can themselves act as prodrugs of other compounds of the present invention. It is also possible that two compounds of the present invention combine to form a prodrug form. In certain situations, the prodrug of the compound of the present invention can be prepared by internally linking two functional groups in the compound of the present invention, for example, by forming a lactone.

[0212] Metabolites Active metabolites of the compounds of the present invention, that is, compounds that are often formed in vivo by oxidation or dealkylation when the drug is administered, are also included within the scope of the present invention. Some examples of metabolites according to the present invention include, but are not limited to: (i) When the compound of the present invention contains an alkyl group, its hydroxyalkyl derivative (-CH > -COH): (ii) When the compound of the present invention contains an alkoxy group, its hydroxy derivative (-OR → -OH); (iii) When the compound of the present invention contains a tertiary amino group, its secondary amino derivative (-NRR’ → -NHR or -NHR’); (iv) When the compound of the present invention contains a secondary amino group, its primary derivative (-NHR → -NH2); (v) When the compound of the present invention contains a phenyl moiety, its phenol derivative (-Ph → -PhOH); (vi) When the compound of the present invention contains an amide group, its carboxylic acid derivative (-CONH2 → COOH); and (vii) When the compound contains a hydroxy or carboxylic acid group, the compound may be metabolized, for example, by conjugation with glucuronic acid to form a glucuronide. Other pathways of conjugation metabolism exist. These pathways are often known as phase II metabolism and include, for example, sulfation or acetylation. Other functional groups such as NH groups can also undergo conjugation.

[0213] Pharmaceutical composition In another embodiment, the present invention includes a pharmaceutical composition. For the purposes of the pharmaceutical composition, the compound itself or its pharmaceutically acceptable salt will simply be referred to as the compound of the present invention.

[0214] "Pharmaceutical composition" refers to a mixture of one or more of the compound of the present invention, or its pharmaceutically acceptable salt, solvate, hydrate or prodrug, as an active ingredient, and at least one pharmaceutically acceptable additive.

[0215] The term "additive" is used herein to describe any component other than the compound of the present invention. The choice of additive will depend largely on factors such as the mode of administration, the effect of the additive on solubility and stability, and the nature of the dosage form.

[0216] As used herein, "additive" includes all and any solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, carriers, diluents, and the like that are physiologically compatible. Examples of additives include one or more of water, saline, phosphate buffer solutions, dextrose, glycerol, ethanol, and the like, and combinations thereof, and may include isotonic agents such as sugars, sodium chloride, or polyalcohols such as mannitol or sorbitol in the composition. Examples of additives also include various organic solvents (such as hydrates and solvates). The pharmaceutical composition may, if desired, contain additional additives such as flavoring agents, binders / binding agents, lubricants, disintegrants, sweetening or flavoring agents, coloring agents or dyes, and the like. For example, for oral administration, tablets containing various additives such as citric acid can be used together with various disintegrants such as starch, alginic acid, and certain complex silicates, and binders such as sucrose, gelatin, and gum arabic. By way of non-limiting example, additives include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycol. In addition, lubricants such as magnesium stearate, sodium lauryl sulfate, and talc are often useful for tableting purposes. Solid compositions of the same type can also be used within filled soft and hard gelatin capsules. Accordingly, non-limiting examples of additives also include lactose or milk sugar and high molecular weight polyethylene glycol. When an aqueous suspension or elixir is desired for oral administration, the active compound therein can be combined with various sweetening or flavoring agents, coloring agents or dyes, and, if desired, emulsifying or suspending agents, together with additional additives such as water, ethanol, propylene glycol, glycerol, or combinations thereof.

[0217] Examples of additives include pharmaceutically acceptable substances that enhance the shelf life or effectiveness of the compound, such as wetting agents or minor auxiliary substances, such as wetting agents or emulsifiers, preservatives, or buffering agents.

[0218] The compositions of the present invention may be in various forms. These include, for example, liquid formulations (e.g., injection and infusion solutions), dispersions or suspensions, tablets, capsules, pills, powders, liposome formulations, and suppositories, etc., including liquid, semi-solid, and solid dosage forms. The form depends on the intended mode of administration and therapeutic application.

[0219] Typical compositions are in the form of injection or infusion solutions, such as those generally used for passive immunization of humans with antibodies. One mode of administration is parenteral (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular). In another embodiment, the compound is administered by intravenous infusion or injection. In yet another embodiment, the compound is administered by intramuscular or subcutaneous injection.

[0220] Oral administration of solid dosage forms can be provided, for example, in separate units such as hard or soft capsules, pills, cachets, lozenges, or tablets, each containing a predetermined amount of at least one compound of the present invention. In another embodiment, oral administration may be in the form of a powder or granules. In another embodiment, the oral dosage form is sublingual, such as a lozenge. In such solid dosage forms, the compounds of the present invention are usually combined with one or more auxiliary agents. Such capsules or tablets may include controlled release formulations. In the case of capsules, tablets, and pills, the dosage form may include a buffering agent or may be prepared with an enteric coating.

[0221] In another embodiment, oral administration may be in a liquid dosage form. Liquid dosage forms for oral administration include, for example, pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art (such as water). Such compositions may contain adjuvants such as one or more of wetting agents, emulsifying agents, suspending agents, flavoring agents (such as sweetening agents), or fragrances.

[0222] In another embodiment, the present invention includes parenteral dosage forms. "Parenteral administration" includes, for example, subcutaneous injection, intravenous injection, intraperitoneal, intramuscular injection, intracardiac injection, and infusion. Injectable preparations (i.e., sterile aqueous or oily suspensions for injection) can be formulated using one or more suitable dispersing agents, wetting agents, or suspending agents according to known techniques.

[0223] In another embodiment, the present invention includes topical dosage forms. "Topical administration" includes, for example, cutaneous and transdermal administration via a transdermal patch or iontophoresis device, intraocular administration, or intranasal or inhalation administration. Compositions for topical administration also include, for example, topical gels, sprays, ointments, and creams. Topical formulations may contain compounds that enhance the absorption or penetration of the active ingredient through the skin or other affected areas. When administering the compounds of the present invention by a transdermal device, administration will be achieved using a patch of either the reservoir and porous membrane type or the solid matrix variant. Typical formulations for this purpose include gels, hydrogels, lotions, solutions, creams, ointments, powders, dressings, foams, films, skin patches, wafers, implants, sponges, fibers, bandages, and microemulsions. Liposomes can also be used. Typical additives include alcohol, water, mineral oil, liquid petrolatum, white petrolatum, glycerin, polyethylene glycol, and propylene glycol. Penetration enhancers can be incorporated. See, for example, B.C. Finnin and T.M. Morgan, J. Pharm. Sci., vol. 88, pp. 955 - 958, 1999.

[0224] Formulations suitable for topical administration to the eye include, for example, eye drops in which the compounds of the present invention are dissolved or suspended in suitable additives. Typical formulations suitable for eye or ear administration may be in the form of drops of a superfine pulverized suspension or solution in isotonic, pH-adjusted, sterile physiological saline. Other formulations suitable for eye and ear administration include ointments, biodegradable (i.e., absorbable gel sponges, collagen) and non-biodegradable (i.e., silicone) implant agents, wafers, lens agents, and particulate or vesicular systems such as niosomes or liposomes. Polymers such as cross-linked polyacrylic acid, polyvinyl alcohol, hyaluronic acid, cellulose-based polymers, for example, hydroxypropylmethylcellulose, hydroxyethylcellulose, or methylcellulose, or heteropolysaccharide polymers, for example, polymers such as gellan gum, can be incorporated together with preservatives such as benzalkonium chloride. Such formulations can also be delivered by iontophoresis.

[0225] For nasal administration, the compounds of the present invention are preferably delivered as a solution or suspension from a pump-type spray container that the patient squeezes or pumps, or as an aerosol spray delivery from a pressurized container or nebulizer using a suitable propellant. Formulations suitable for nasal administration are typically in the form of a dry powder from a dry powder inhaler (either alone or as a mixture, for example, as a dry blend with lactose or as mixed constituent particles, for example, mixed with a phospholipid such as phosphatidylcholine), or as an aerosol spray from a pressurized container, pump, spray, atomizer (preferably an atomizer that uses electrohydrodynamics to produce a fine mist), or nebulizer with or without a suitable propellant such as 1,1,1,2-tetrafluoroethane or 1,1,1,2,3,3,3-heptafluoropropane. For nasal use, the powder may contain a bioadhesive, for example, chitosan or cyclodextrin.

[0226] In another embodiment, the present invention includes a rectal dosage form. Such a rectal dosage form may be, for example, in the form of a suppository. Although cocoa butter is a conventional suppository base, various alternatives can be used if appropriate.

[0227] Other additives and modes of administration known in the pharmaceutical art can also be used. The pharmaceutical compositions of the present invention can be prepared by any of the well-known techniques of pharmacy, such as effective formulation and administration procedures. The above considerations regarding effective formulation and administration procedures are well known in the art and are described in standard textbooks. The formulation of drugs is discussed, for example, in Hoover, John E., Remington’s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania, 1975; Liberman et al., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y., 1980; and Kibbe et al., Handbook of Pharmaceutical Excipients (Third Edition), American Pharmaceutical Association, Washington, 1999.

[0228] Acceptable additives are non-toxic to the subject at the dosages and concentrations used and may include one or more of the following: 1) buffers such as phosphates, citrates, and other organic acids; 2) salts such as sodium chloride; 3) antioxidants such as ascorbic acid and methionine; 4) preservatives such as octadecyl dimethyl benzyl ammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl or benzyl alcohol; 5) alkyl parabens such as methyl or propyl paraben, catechol, resorcinol, cyclohexanol, 3-pentanol, or m-cresol; 6) low molecular weight (less than about 10 residues) polypeptides; 7) proteins such as serum albumin, gelatin, or immunoglobulins; 8) hydrophilic polymers such as polyvinyl pyrrolidone; 9) amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; 10) monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrin; 11) chelating agents such as EDTA; 12) sugars such as sucrose, mannitol, trehalose or sorbitol; 13) salt-forming counterions such as sodium, metal complexes (e.g., Zn-protein complexes), or 14) nonionic surfactants such as polysorbate (e.g., polysorbate 20 or polysorbate 80), poloxamer or polyethylene glycol (PEG).

[0229] For oral administration, the composition can be provided in the form of tablets or capsules containing 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 75.0, 100, 125, 150, 175, 200, 250, 500 or 1000 milligrams of the active ingredient for symptomatic adjustment of the dosage to the patient. The medicament typically contains from about 0.01 mg to about 500 mg of the active ingredient, or in another embodiment, from about 1 mg to about 100 mg of the active ingredient. Intravenously, the dosage may range from about 0.01 to about 10 mg / kg / min during a constant rate infusion.

[0230] Liposomes containing the compounds of the present invention can be prepared by methods known in the art (see, for example, Chang, H.I.; Yeh, M.K.; Clinical development of liposome-based drugs: formulation, characterization, and therapeutic efficacy; Int J Nanomedicine 2012;7;49 - 60). Particularly useful liposomes can be produced by the reverse phase evaporation method using a lipid composition comprising phosphatidylcholine, cholesterol, and a PEG - derivatized phosphatidylethanolamine (PEG - PE). The liposomes are extruded through a filter of a defined pore size to obtain liposomes having a desired diameter.

[0231] The compounds of the present invention can also be encapsulated, for example, by coacervation techniques or in colloidal drug delivery systems (such as liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules) or in macroemulsions in microcapsules prepared by interfacial polymerization, such as hydroxymethylcellulose or gelatin - microcapsules and poly - (methyl methacrylate) microcapsules. Such techniques are disclosed in Remington, The Science and Practice of Pharmacy, 20th Edition, Mack Publishing (2000).

[0232] Sustained release preparations can be used. Suitable examples of sustained release preparations include semipermeable matrices of solid hydrophobic polymers containing the compounds of the present invention, and the matrices are in the form of molded articles such as films or microcapsules. Examples of sustained release matrices include polyesters, hydrogels (e.g., poly(2-hydroxyethyl-methacrylate) or poly(vinyl alcohol)), polylactides, copolymers of L-glutamic acid and 7-ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as those used in leuprolide acetate for depot suspensions (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), sucrose acetate isobutyrate, and poly-D-(-)-3-hydroxybutyric acid.

[0233] Formulations for intravenous administration must be sterile. This can be readily achieved, for example, by filtration through a sterile filtration membrane. The compounds of the present invention are generally placed in a container having a sterile access port, such as an intravenous fluid bag or vial having a stopper that can be penetrated by a hypodermic needle.

[0234] Suitable emulsions can be prepared using commercially available lipid emulsions, such as lipid emulsions containing soybean oil, lipid emulsions for intravenous administration (e.g., containing safflower oil, soybean oil, egg phospholipid, and glycerin in water), emulsions containing soybean oil and medium-chain triglycerides, and lipid emulsions of cottonseed oil. The active ingredient can be dissolved in a pre-mixed emulsion composition or, alternatively, dissolved in an emulsion formed by mixing an oil (e.g., soybean oil, safflower oil, cottonseed oil, sesame oil, corn oil, or almond oil) and a phospholipid (e.g., egg phospholipid, soybean phospholipid, or soybean lecithin) with water. It will be appreciated that other components, such as glycerol or glucose, can be added to adjust the tonicity of the emulsion. Suitable emulsions should typically contain up to 20%, for example, between 5 - 20% oil. The lipid emulsion contains fat droplets between 0.1 - 1.0 μm, particularly between 0.1 - 0.5 μm, and can have a pH in the range of 5.5 - 8.0.

[0235] For example, the emulsion composition can be prepared by mixing the compound of the invention with a lipid emulsion containing soybean oil or its constituents (soybean oil, egg phospholipid, glycerol, and water).

[0236] Compositions for inhalation or insufflation include solutions and suspensions, as well as powders, in a pharmaceutically acceptable aqueous or organic solvent, or mixtures thereof. The liquid or solid composition may contain suitable pharmaceutically acceptable additives as presented above. In some embodiments, the composition is administered orally or via the nasal respiratory route for local or systemic action. Preferably, the composition in a sterile pharmaceutically acceptable solvent can be atomized by the use of a gas. The atomized solution can be breathed directly from the atomizing device or the atomizing device can be attached to a face mask, tent, or intermittent positive pressure ventilator. The solution, suspension, or powder composition can be administered preferably orally or nasally from a device that delivers the formulation in a suitable manner.

[0237] A formulation intermediate (DPI) is a partially processed substance that needs to be subjected to further processing steps before it becomes a bulk formulation. The compounds of the present invention can be formulated into a formulation intermediate DPI that contains the active ingredient in a higher free energy form than the crystalline form. One reason for using DPI is to improve the characteristics of oral absorption due to low solubility, slow dissolution, improved substance transport through the mucin layer adjacent to epithelial cells, and in some cases, limitations caused by biological barriers such as metabolism and transporters. Other reasons can include improved solid state stability and downstream manufacturability. In one embodiment, the formulation intermediate contains the compound of the present invention isolated and stabilized in an amorphous state (e.g., an amorphous solid dispersion (ASD)). There are many techniques known in the art for producing an ASD that produces a substance suitable for incorporation into bulk formulations such as spray dried dispersions (SDD), melt extrudates (often referred to as HME), co-precipitates, amorphous drug nanoparticles, and nano-adsorbates. In one embodiment, the amorphous solid dispersion contains the compound of the present invention and a polymer additive. Other additives, as well as the concentrations of said additives and the compound of the present invention, are well known in the art and are described in standard textbooks. See, for example, "Amorphous Solid Dispersions Theory and Practice" by Navnit Shah et al.

[0238] Administration and Dosage As used herein, the terms "treating," "treat," or "treatment" include both prophylactic, i.e., protective, and palliative treatment, i.e., reducing, alleviating, or slowing the progression of a patient's disease (or condition) or any tissue damage associated with the disease.

[0239] As used herein, the terms "subject", "individual" or "patient", used interchangeably, refer to any animal, including mammals. Mammals according to the invention include dogs, cats, cows, goats, horses, sheep, pigs, rodents, rabbits, primates, humans and the like, including mammals in utero. In one embodiment, a human is a preferred subject. A human subject can be of either sex and at any stage of development.

[0240] As used herein, the phrase "therapeutically effective amount" refers to the amount of an active compound or pharmaceutical agent that elicits a biological or medical response in a tissue, system, animal, individual or human, as determined by a researcher, veterinarian, physician or other clinician, and includes one or more of the following: (1) Preventing a disease; for example, preventing a disease, condition or disorder in an individual who is susceptible to the disease, condition or disorder but has not yet experienced or manifested the pathology or overall symptoms; (2) Inhibiting a disease; for example, inhibiting a disease, condition or disorder in an individual who is experiencing or manifesting the pathology or overall symptoms of the disease, condition or disorder (i.e., preventing (or slowing) further development of the pathology or overall symptoms or both); and (3) Remitting a disease; for example, remitting a disease, condition or disorder in an individual who is experiencing or manifesting the pathology or overall symptoms of the disease, condition or disorder (i.e., reversing the pathology or overall symptoms or both).

[0241] Typically, the compounds of the invention are administered in an amount effective to treat the conditions as described herein. The compounds of the invention can be administered as the compound itself or alternatively as a pharmaceutically acceptable salt. For purposes of administration and dosing, the compound itself or its pharmaceutically acceptable salt will simply be referred to as the compounds of the invention.

[0242] The compounds of the present invention are administered by any suitable route, in the form of a pharmaceutical composition adapted to such a route, and in a dosage effective for the intended treatment. The compounds of the present invention can be administered orally, rectally, vaginally, parenterally, topically, intranasally, or by inhalation.

[0243] The compounds of the present invention can be administered orally. Oral administration may involve swallowing such that the compound enters the gastrointestinal tract, or buccal or sublingual administration such that the compound enters the bloodstream directly from the mouth.

[0244] In another embodiment, the compounds of the present invention may be administered parenterally, for example, directly into the bloodstream, muscle, or viscera. Suitable means for parenteral administration include intravenous, intraarterial, intraperitoneal, intrathecal, intracerebroventricular, intraurethral, intrasternal, intracranial, intramuscular, and subcutaneous. Suitable devices for parenteral administration include needles (including micro needles) syringes, needleless syringes, and infusion techniques.

[0245] In another embodiment, the compounds of the present invention can also be administered topically to the skin or mucosa, i.e., to the skin or transdermally. In another embodiment, the compounds of the present invention can be administered intranasally or by inhalation. In another embodiment, the compounds of the present invention can be administered rectally or vaginally. In another embodiment, the compounds of the present invention can also be administered directly to the eye or ear.

[0246] The dosing regimen with the compounds of the present invention or compositions containing said compounds is based on a variety of factors including the species, age, weight, sex and medical condition of the patient; the severity of the condition; the route of administration; and the activity of the particular compound used. Accordingly, the dosing regimen can vary widely. In one embodiment, the total daily dose of the compounds of the present invention is typically about 0.01 to about 100 mg / kg (i.e., mg of the compound of the present invention per kg of body weight) for the treatment of the indication conditions discussed herein. In another embodiment, the total daily dose of the compounds of the present invention is about 0.1 to about 50 mg / kg, and in another embodiment, about 0.5 to about 30 mg / kg. It is not uncommon to repeat the administration of the compounds of the present invention multiple times (typically within 4 times) per day. If desired, multiple doses per day can typically be used to increase the total daily dose.

[0247] Methods of treatment and uses The compounds of the present invention can inhibit all the activities of the KRAS G12C, KRAS G12D, and KRAS G12V receptors and can be useful in the treatment, prevention, suppression, and remission of diseases such as cancers, disorders, and pathologies mediated by any of the KRAS G12C, KRAS G12D, and KRAS G12V receptors, or combinations thereof.

[0248] Cancers to be treated include squamous cell carcinoma, basal cell carcinoma, myeloma, small cell lung cancer, non-small cell lung cancer, glioma, Hodgkin lymphoma, non-Hodgkin lymphoma, acute myeloid leukemia (AML), multiple myeloma, gastrointestinal (tract) cancer, kidney cancer, ovarian cancer, liver cancer, lymphoblastic leukemia, lymphocytic leukemia, colorectal cancer, endometrial cancer, kidney cancer, prostate cancer, thyroid cancer, melanoma, chondrosarcoma, neuroblastoma, pancreatic cancer, glioblastoma multiforme, cervical cancer, brain tumor, gastric cancer, uterine cancer, bladder cancer including muscle-invasive non-invasive bladder cancer, hepatocellular carcinoma, breast cancer, and head and neck cancer.

[0249] Preferably, the compounds of the present invention may be useful for treating lung cancer such as non-small cell lung cancer (NSCLC), pancreatic cancer, colorectal cancer, breast cancer, blood cancer, gynecological cancer, prostate cancer, or skin cancer. See Mustachio, L., Targeting KRAS in Cancer: Promising Therapeutic Strategies, Cancers, 2021, 13, 1204.

[0250] More preferably, the compounds of the present invention may be useful for treating non-small cell lung cancer (NSCLC), pancreatic cancer, and colorectal cancer.

[0251] Co-administration The compounds of the present invention can be used alone or in combination with one or more other therapeutic agents. The present invention provides any of the uses, methods or compositions as defined herein, which use the compounds of the present invention, or pharmaceutically acceptable salts thereof, in combination with one or more other therapeutic anti-cancer drugs described herein.

[0252] The administration of two or more compounds "in combination" means administering all of the compounds in close temporal proximity to affect the treatment of the subject. Two or more compounds can be administered simultaneously or sequentially, with or without specific time limits, via the same or different routes of administration, according to the treatment regimen, at the same or different dosing schedules. In addition, simultaneous administration can be effected by mixing the compounds prior to administration or by administering the compounds at the same time but in separate dosage forms at the same or different sites of administration. Examples of "in combination" include, but are not limited to, "co-administration", "simultaneous administration", "synchronous administration", "sequential administration" and "administered synchronously".

[0253] The compounds of the present invention and one or more other therapeutic agents can be administered as a fixed or non-fixed combination of active ingredients. The term "fixed combination" means that both the compound of the present invention, or a pharmaceutically acceptable salt thereof, and one or more therapeutic agents are administered to a subject synchronously in a single composition or dosage. The term "non-fixed combination" means that the compound of the present invention, or a pharmaceutically acceptable salt thereof, and one or more therapeutic agents are formulated as separate compositions or dosages and are administered to a subject in need thereof synchronously or at different times with a variable intervening time period such that such administration can result in effective levels of two or more compounds in the subject's body.

[0254] The group of additional chemotherapeutic agents that can be administered in combination with the compounds of the present invention includes, but is not limited to: alkylating agents, antimetabolites, kinase inhibitors, spindle inhibitor plant alkaloids, cytotoxic / antitumor antibiotics, topoisomerase inhibitors, photosensitizers, antiestrogens and selective estrogen receptor modulators (SERMs), antiprogesterones, estrogen receptor downregulators (ERDs), estrogen receptor antagonists, luteinizing hormone releasing hormone agonists; IL-2 receptor agonists (recombinant cytokines or agonists for cytokine receptors); and antisense oligonucleotides or oligonucleotide derivatives that inhibit the expression of genes involved in abnormal cell proliferation or tumor growth.

[0255] Other additional chemotherapeutic agents include not only taxanes or platinum formulations, but also HER2-targeted drugs, such as trastuzumab.

[0256] In another embodiment, such additional anti-cancer therapeutic agents include compounds derived from the following groups: mitotic inhibitors, alkylating agents, antimetabolites, antitumor antibiotics, antiangiogenic agents, topoisomerase I and II inhibitors, plant alkaloids, spindle-inhibiting plant alkaloids, MCT4 inhibitors; MAT2a inhibitors; alk / c-Met / ROS inhibitors (including crizotinib or lorlatinib); mTOR inhibitors (including temsirolimus or gedatolisib); src / abl inhibitors (including bosutinib); cyclin-dependent kinase (CDK) inhibitors (including palbociclib, PF-06873600); erb inhibitors (including dacomitinib); PARP inhibitors (including talazoparib); SMO inhibitors (including glasdegib); EGFR T790M inhibitors; PRMT5 inhibitors; TGFβR1 inhibitors; growth factor inhibitors; cell cycle inhibitors, biological response modifiers; enzyme inhibitors; and cytotoxic agents.

[0257] In another embodiment, such additional anti-cancer therapeutic agents include, for example, anti-angiogenic agents (including sunitinib, axitinib, sorafenib, and tivozanib) that include tyrosine kinase / vascular endothelial growth factor (VEGF) receptor (VEGFR) inhibitors, TIE-2 inhibitors, PDGFR inhibitors, angiopoietin inhibitors, PKCβ inhibitors, COX-2 (cyclooxygenase II) inhibitors, integrin (alpha-v / beta-3), MMP-2 (matrix-metalloproteinase 2) inhibitors, and compounds derived from MMP-9 (matrix-metalloproteinase 9) inhibitors. Preferred anti-angiogenic agents include sunitinib (Sutent (trademark)), bevacizumab (Avastin (trademark)), axitinib (Inlyta (trademark)), SU14813 (Pfizer), and AG13958 (Pfizer). Additional anti-angiogenic agents include batatinib (CGP79787), pegaptanib octasodium (Macugen (trademark)), vandetanib (Zactima (trademark)), PF-0337210 (Pfizer), SU14843 (Pfizer), AZD2171 (AstraZeneca), ranibizumab (Lucentis (trademark)), Neovastat (trademark) (AE941), tetrathiomolybdata (Coprexa (trademark)), AMG706 (Amgen), VEGF Trap (AVE0005), CEP7055 (Sanofi-Aventis), XL880 (Exelixis), teratinib (BAY57-9352), and CP-868,596 (Pfizer). Other anti-angiogenic agents include enzastaurin (LY317615), midostaurin (CGP41251), perifosine (KRX0401), teprenone (Selbex (trademark)) and UCN 01 (Kyowa Hakko).Other examples of anti-angiogenic drugs include celecoxib (Celebrex™), parecoxib (Dynastat™), deracoxib (SC59046), lumiracoxib (Preige™), valdecoxib (Bextra™), rofecoxib (Vioxx™), iguratimod (Careram™), IP751 (Invedus), SC-58125 (Pharmacia), and etoricoxib (Arcoxia™). Still further anti-angiogenic drugs include exisulind (Aptosyn™), salsalate (Amigesic™), diflunisal (Dolobid™), ibuprofen (Motrin™), ketoprofen (Orudis™), nabumetone (Relafen™), piroxicam (Feldene™), naproxen (Aleve™, Naprosyn™), diclofenac (Voltaren™), indomethacin (Indocin™), sulindac (Clinoril™), tolmetin (Tolectin™), etodolac (Lodine™), ketorolac (Toradol™), and oxaprozin (Daypro™). Still further anti-angiogenic drugs include ABT510 (Abbott), aplastat (TMI005), AZD8955 (AstraZeneca), incyclinide (Metastat™), and PCK3145 (Procyon).Additional anti-angiogenic agents further include acitretin (Neotigason (trademark)), plitidepsin (aplidine (trademark)), siramesine (EMD121974), combretastatin A4 (CA4P), fenretinide (4HPR), halofuginone (Tempostatin (trademark)), Panzem (trademark) (2-methoxyestradiol), PF-03446962 (Pfizer), rebimastat (BMS275291), catumaxomab (Removab (trademark)), lenalidomide (Revlimid (trademark)), squalamine (EVIZON (trademark)), thalidomide (Thalomid (trademark)), Ukrain (trademark) (NSC631570), Vitaxin (trademark) (MEDI522), and zoledronic acid (Zometa (trademark)).

[0258] In another embodiment, such additional anti-cancer therapeutic agents include compounds derived from hormonal drugs and antagonists. Examples include anti-hormonal drugs that act to modulate or inhibit the hormonal action on tumors, for example, anti-estrogens and selective estrogen receptor modulators (SERMs), as well as selective estrogen receptor degraders (SERDs), which include tamoxifen, raloxifene, droloxifene, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, toremifene (Fareston), and fulvestrant. Examples also include aromatase inhibitors that inhibit the enzyme aromatase that regulates estrogen production in the adrenal glands, 4(5)-imidazole, aminoglutethimide, megestrol acetate, exemestane, formestane, fadrozole, vorozole, letrozole, and anastrozole; and compounds such as anti-androgens like flutamide, nilutamide, bicalutamide, leuprolide, flururidyl, apalutamide, enzalutamide, cimetidine, and goserelin.

[0259] In another embodiment, such additional anti-cancer therapeutic agents include signal transduction inhibitors such as inhibitors of protein tyrosine kinases and / or serine / threonine kinases: compounds derived from signal transduction inhibitors (e.g., inhibiting the means by which regulatory molecules govern the basic processes of cell proliferation, differentiation, and survival that are transmitted intracellularly). Signal transduction inhibitors include small molecules, antibodies, and antisense molecules. Signal transduction inhibitors include, for example, kinase inhibitors (e.g., tyrosine kinase inhibitors or serine / threonine kinase inhibitors) and cell cycle inhibitors. More specifically, signal transduction inhibitors include, for example, farnesyl protein transferase inhibitors, EGF inhibitors, ErbB-1 (EGFR), ErbB-2, pan erb, IGF1R inhibitors, MEK (including binimetinib (Mektovi (trademark))), c-Kit inhibitors, FLT-3 inhibitors, K-Ras inhibitors, PI3 kinase inhibitors, JAK inhibitors, STAT inhibitors, Raf kinase inhibitors, BRAF (including encorafenib (Braftovi (trademark))), Akt inhibitors, mTOR inhibitors, P70S6 kinase inhibitors, inhibitors of the WNT pathway, and multi-target kinase inhibitors.

[0260] In another embodiment, such additional anti-cancer therapeutic agents include docetaxel, paclitaxel, paclitaxel protein-bound particles, cisplatin, carboplatin, oxaliplatin, capecitabine, gemcitabine, or vinorelbine.

[0261] In another embodiment, such additional anti-cancer therapeutic agents include compounds derived from epigenetic modulators, where examples include inhibitors of EZH2 (including PF-06821497), SMARCA4, PBRM1, ARID1A, ARID2, ARID1B, DNMT3A, TET2, MLL1 / 2 / 3, NSD1 / 2, SETD2, BRD4, DOT1L, HKMTsanti, PRMT1-9, LSD1, UTX, IDH1 / 2, or BCL6.

[0262] In another embodiment, such additional anti-cancer therapeutic agents include compounds that are cancer immunotherapeutics, including immunomodulatory agents.

[0263] In another embodiment, combinations with pattern recognition receptors (PRRs) are contemplated. PRRs are receptors expressed by cells of the immune system that recognize various molecules associated with pathogens and / or cell damage or death. PRRs are involved in both innate and adaptive immune responses. PRR agonists can be used to stimulate an immune response in a subject. There are multiple groups of PRR molecules, including toll-like receptors (TLRs), RIG-I-like receptors (RLRs), nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs), C-type lectin receptors (CLRs), and stimulator of interferon genes (STING) proteins.

[0264] The STING protein functions as both a cytosolic DNA sensor and an adapter protein in type I interferon signaling. The terms "STING" and "stimulator of interferon genes" refer to any form of the STING protein, as well as variants, isoforms, and species homologs that maintain at least a portion of the activity of STING. Unless otherwise indicated, such as by a specific reference to human STING, STING includes all mammalian species of the native sequence STING, for example, human, monkey, and mouse STING, which is also known as TMEM173.

[0265] As used herein, a "STING agonist" means any molecule that, when bound to STING, (1) stimulates or activates STING, (2) enhances, increases, promotes, induces, or prolongs the activity, function, or presence of STING, or (3) enhances, increases, promotes, or induces the expression of STING. STING agonists useful in any of the treatment methods, medicaments, and uses of the present invention include, for example, nucleic acid ligands that bind to STING.

[0266] Examples of STING agonists useful in the treatment methods, medicaments and uses of the present invention include various immunostimulatory nucleic acids such as synthetic double-stranded DNA, cyclic di-GMP, cyclic-GMP-AMP (cGAMP), synthetic cyclic dinucleotides (CDNs) such as MK-1454 and ADU-S100 (MIW815), and small molecules such as WO2019027858, WO20180093964, WO2017175156, WO2017175147.

[0267] Therapeutic antibodies can have specificity for a variety of different antigens. For example, a therapeutic antibody can be directed against a tumor-associated antigen, and binding of the antibody to the antigen promotes the death of cells expressing the antigen. In other examples, a therapeutic antibody can be directed against an antigen on an immune cell, and binding of the antibody prevents downregulation of the activity of cells expressing the antigen (and thereby promotes the activity of cells expressing the antigen). In some situations, a therapeutic antibody can function via multiple different mechanisms (e.g., both i) promoting the death of cells expressing the antigen and ii) preventing the antigen from causing downregulation of the activity of immune cells in contact with cells expressing the antigen are possible).

[0268] In another embodiment, such additional anti-cancer therapeutics include antibodies that would be blocking or inhibitory at the target: CTLA-4 (including ipilimumab or tremelimumab), PD-1 or PD-L1 (including atezolizumab, avelumab, semipilumab, durvalumab, nivolumab, sasanlimab, or pembrolizumab), LAG-3, TIM-3, or TIGIT.

[0269] In another embodiment, such additional anti-cancer therapeutics include antibodies that are agonists of 4-1BB, OX40, GITR, ICOS, or CD40.

[0270] In another embodiment, the anti-cancer treatment may be CAR-T-cell treatment.

[0271] Examples of therapeutic antibodies include anti-OX40 antibodies, anti-4-1BB antibodies, anti-HER2 antibodies (including anti-HER2 antibody-drug conjugates (ADCs)), bispecific anti-CD47 / anti-PD-L1 antibodies, and bispecific anti-P-cadherin / anti-CD3 antibodies. Examples of cytotoxic agents that can be incorporated into ADCs include anthracyclines, auristatins, dolastatin, combretastatin, duocarmycin, pyrrolobenzodiazepine dimers, indolino-benzodiazepine dimers, enediyne, geldanamycin, maytansine, puromycin, taxanes, vinca alkaloids, camptothecin, tubulysin, hemiasterlin, spirostatin, pladienolide, and their stereoisomers, homologs, analogs, or derivatives. Exemplary immunomodulatory agents that can be incorporated into ADCs include ganciclovir, etanercept, tacrolimus, sirolimus, pimecrolimus, cyclosporine, rapamycin, cyclophosphamide, azathioprine, mycophenolate mofetil, methotrexate, glucocorticoids and their analogs, cytokines, stem cell growth factors, lymphotoxin, tumor necrosis factor (TNF), hematopoietic factors, interleukins (e.g., interleukin-1 (IL-1), IL-2, IL-3, IL-6, IL-10, IL-12, IL-15, IL-18, and IL-21), colony-stimulating factors (e.g., granulocyte-colony stimulating factor (G-CSF) and granulocyte macrophage-colony stimulating factor (GM-CSF)), interferons (e.g., interferon-alpha, -beta and -gamma), a stem cell growth factor called "S1 factor", erythropoietin and thrombopoietin, or combinations thereof.

[0272] Additional examples of therapeutic antibodies can include the following antigens, and exemplary antibodies directed against such antigens are also included below (within the brackets / parentheses after the antigen). The following antigens may also be referred to herein as "target antigens" and the like. Target antigens for therapeutic antibodies herein include, for example: 4-1BB (e.g., utomilumab); 5T4; A33; alpha-folate receptor 1 (e.g., mirvetuximab soravtansine); Alk-1; BCMA [see, e.g., US9969809]; BTN1A1 (see, e.g., WO2018222689); CA-125 (e.g., abagomomab); carbonic anhydrase IX; CCR2; CCR4 (e.g., mogamulizumab); CCR5 (e.g., leronlimab); CCR8; CD3 [e.g., blinatumomab (CD3 / CD19 bispecific), CD3 / P-cadherin bispecific, CD3 / BCMA bispecific] CD19 (e.g., blinatumomab, MOR208); CD20 (e.g., ibritumomab tiuxetan, obinutuzumab, ofatumumab, rituximab, ublituximab); CD22 (inotuzumab ozogamicin, moxetumomab pasudotox); CD25; CD28; CD30 (e.g., brentuximab vedotin); CD33 (e.g., gemtuzumab ozogamicin); CD38 (e.g., daratumumab, isatuximab), CD40; CD-40L; CD44v6; CD47 (e.g., Hu5F9-G4, CC-90002, SRF231, B6H12); CD52 (e.g., alemtuzumab); CD56; CD63; CD79 (e.g., polatuzumab vedotin); CD80; CD123; CD276 / B7-H3 (e.g., omburtamab); CDH17; CEA; ClhCG; CTLA-4 (e.g., ipilimumab, tremelimumab), CXCR4; desmoglein 4; DLL3 (e.g., lobaplizumab tesirine); DLL4; E-cadherin; EDA; EDB; EFNA4; EGFR (e.g., cetuximab, depatuxizumab mafodotin, necitumumab, panitumumab); EGFRvIII; endosialin; EpCAM (e.g., oportuzumab monatox); FAP; fetal acetylcholine receptor;FLT3 (see, e.g., WO2018 / 220584); GD2 (e.g., dinutuximab, 3F8); GD3; GITR; GloboH; GM1; GM2; HER2 / neu [e.g., margetuximab, pertuzumab, trastuzumab; ado-trastuzumab emtansine, trastuzumab duocarmazine, see [US8828401]; HER3; HER4; ICOS; IL-10; ITG-AvB6; LAG-3 (e.g., relatlimab); Lewis-Y; LG; Ly-6; M-CSF (see [US7326414]); MCSP; mesothelin; MUC1; MUC2; MUC3; MUC4; MUC5AC; MUC5B; MUC7; MUC16; Notch1; Notch3; nectin-4 (e.g., enfortumab vedotin); OX40 (see [US7960515]); P-cadherin (see [WO2016 / 001810]); PCDHB2; PDGFRA (e.g., olaparib); plasma cell antigen; polySA; PSCA; PSMA; PTK7 (see [US9409995]); Ror1; SAS; SCRx6; SLAMF7 (e.g., elotuzumab); SHH; SIRPa (e.g., ED9, Effi-DEM); STEAP; TGF-beta; TIGIT; TIM-3; TMPRSS3; TNF-alpha precursor; TROP-2 (e.g., sacituzumab govitecan); TSPAN8; VEGF (e.g., bevacizumab, brolucizumab); VEGFR1 (e.g., ranibizumab); VEGFR2 (e.g., ramucirumab, ranibizumab); Wue-1 is included.;

[0273] Exemplary imaging agents that may be included in the ADC include fluorescein, rhodamine, lanthanide phosphors, and their derivatives, or radioisotopes bound to chelating agents. Examples of fluorophores include, but are not limited to, fluorescein isothiocyanate (FITC) (e.g., 5-FITC), fluorescein amidite (FAM) (e.g., 5-FAM), eosin, carboxyfluorescein, erythrosin, Alexa Fluor® (e.g., Alexa350, 405, 430, 488, 500, 514, 532, 546, 555, 568, 594, 610, 633, 647, 660, 680, 700, or 750), carboxytetramethylrhodamine (TAMRA) (e.g., 5,-TAMRA), tetramethylrhodamine (TMR), and sulforhodamine (SR) (e.g., SR101). Examples of chelating agents include, but are not limited to, 1,4,7,10-tetraazacyclododecane-N,N’,N”,N”’-tetraacetic acid (DOTA), 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA), 1,4,7-triazacyclononane, 1-glutaric acid-4,7-acetic acid (deferoxamine), diethylenetriaminepentaacetic acid (DTPA), and 1,2-bis(o-aminophenoxy)ethane-N,N,N’,N’-tetraacetic acid) (BAPTA).

[0274] Exemplary therapeutic proteins that may be included in the ADC include toxins, hormones, enzymes, and growth factors.

[0275] Exemplary biocompatible polymers that may be incorporated into the ADC include water-soluble polymers such as polyethylene glycol (PEG) or its derivatives, and zwitterion-containing biocompatible polymers (e.g., phosphorylcholine-containing polymers).

[0276] Exemplary biocompatible polymers that may be incorporated into the ADC include antisense oligonucleotides.

[0277] The present invention also relates to the use of radiation in combination with any anti-cancer therapeutic agent administered herein. More specifically, the compounds of the present invention can be administered in combination with additional treatments such as radiotherapy and / or chemotherapy.

[0278] These agents and the compounds of the present invention can be combined with pharmaceutically acceptable vehicles such as saline, Ringer's solution, dextrose solutions, and the like. Specific dosing regimens, i.e., dosage, timing, and frequency, will depend on the particular individual and the medical history of that individual.

[0279] Kit Another aspect of the present invention provides a kit comprising a compound of the present invention or a pharmaceutical composition comprising a compound of the present invention. The kit may further comprise a diagnostic or therapeutic agent in addition to the compound of the present invention or its pharmaceutical composition. The kit may also comprise instructions for use in a diagnostic or therapeutic method. In some embodiments, the kit comprises a compound or its pharmaceutical composition and a diagnostic agent. In other embodiments, the kit comprises a compound or its pharmaceutical composition and one or more therapeutic agents.

[0280] In yet another embodiment, the present invention comprises a kit suitable for use in performing the treatment methods described herein. In one embodiment, the kit contains a first dosage form comprising an amount of one or more compounds of the present invention sufficient to carry out the method of the present invention. In another embodiment, the kit comprises an amount of one or more compounds of the present invention sufficient to carry out the method of the present invention, a container for dosage, and a container for dosage.

[0281] Synthesis method The compounds of the present invention can be synthesized by synthetic routes including processes similar to those well-known in the chemical art, in particular in light of the descriptions contained herein. The starting materials are generally available from commercial suppliers or can be prepared using methods well-known to those skilled in the art. Many of the compounds used herein are related to or can be derived from compounds for which one or more of scientific importance or commercial necessity arise. Thus, such compounds may be one or more of 1) those that are commercially available; 2) those reported in the literature, or 3) those prepared by one skilled in the art from other commercially available substances using materials reported in the literature.

[0282] For purposes of illustration, the reaction schemes shown below provide promising routes for synthesizing the compounds of the present invention and also important intermediates. For a more detailed description of the individual reaction steps, please refer to the Examples section below. Those skilled in the art will appreciate that the compounds of the present invention can be synthesized using other synthetic routes. Although specific starting materials and reagents are discussed below, other starting materials and reagents can be substituted to obtain one or more of various derivatives or reaction conditions. In addition, many of the compounds prepared by the methods below can be further modified using conventional chemical operations well-known to those skilled in the art in light of the present disclosure.

[0283] Those skilled in the art will appreciate that the experimental conditions described in the schemes below are examples of suitable conditions for performing the indicated transformations and that it may be necessary or desirable to vary the exact conditions used to prepare the compounds of the present invention. Further, it will be appreciated that it may be necessary or desirable to perform the transformations in an order different from that described in the schemes or to modify one or more of the transformations to obtain the desired compounds of the present invention.

[0284] In the preparation of the compounds of the present invention, note that some of the preparative methods useful for preparing the compounds described herein may require protection of remote functional groups (e.g., primary amines, secondary amines, carboxyls, etc. in the precursors of the compounds of the present invention). The need for such protection should vary depending on the nature of the remote functional group and the conditions of its preparative method. One of ordinary skill in the art can readily determine the need for such protection. The use of such protection / deprotection methods is also within the scope of skills in the art. For an overview of protecting groups and their use, see March’s Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 8th Edition.

[0285] For example, if a compound contains an amine or carboxylic acid functional group, such a functional group, if left unprotected, may interfere with reactions at other sites of the molecule. Thus, such a functional group can be protected by an appropriate protecting group (PG) that can be removed in a subsequent step. Suitable protecting groups for amine and carboxylic acid protection include those commonly used in peptide synthesis (for amines, N-tert-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz), and 9-fluorenylmethyleneoxycarbonyl (Fmoc) and for carboxylic acids, lower alkyl or benzyl esters, etc.), which are generally not chemically reactive under the described reaction conditions and can typically be removed without chemically altering other functional groups in the compounds of the present invention.

[0286] General Experimental Details 1 H and 19 1H and 19F nuclear magnetic resonance (NMR) spectra were recorded on a Bruker XWIN-NMR (400 or 700 MHz) spectrometer. 1 H and 19 1H and 19F resonances are reported in parts per million (ppm) from tetramethylsilane downfield. 11H NMR data are reported as multiplicities (e.g., s, singlet; d, doublet; t, triplet; q, quartet; quint, quintet; dd, doublet of doublets; dt, doublet of triplets; br s, broad singlet). For spectra obtained in CDCl3, DMSO-d6, and CD3OD, the residual protons (7.27, 2.50, and 3.31 ppm, respectively) were used as internal standards. All observed coupling constants, J, are reported in Hertz (Hz). Exchangeable protons are not always observed.

[0287] Optical rotations were determined on a Jasco P-2000 or Rudolph Autopol IV polarimeter. All final compounds were purified to a purity of ≧95% unless otherwise specified. When the absolute stereochemistry is known, the (R,S) designations are used. When the absolute stereochemistry is not known, the names generated by the software are modified to include the prefixes (+)- and (-)- according to the optical rotation, and the (R * / S * ) designations are used to indicate the relative configuration.

[0288] Mass spectra, MS (m / z), were recorded using either electrospray ionization (ESI) or atmospheric pressure chemical ionization (APCI). When relevant, the m / z data presented are for 19 F, 35 Cl, 79 Br and 127 I, unless otherwise stated.

[0289] The naming is generated and written within PerkinElmer's ChemDraw 18.0.0.231 as described by the IUPAC (International Union of Pure and Applied Chemistry). The naming rules provided by PerkinElmer's ChemDraw 18.0.0.231 are well-known to those skilled in the art, and the naming rules provided by PerkinElmer's ChemDraw 18.0.0.231 are considered to be generally in line with the recommendations of the IUPAC (International Union for Pure and Applied Chemistry) for organic chemical nomenclature and the CAS Index rules.

[0290] Abbreviations aq is aqueous; Bn is benzyl; Boc is tert-butoxycarbonyl; Boc2O is di-tert-butyl dicarbonate; br is broad; tBu is tert-butyl; °C is Celsius temperature; CDCl3 is deuterated chloroform; δ is chemical shift; d is doublet; dd is doublet of doublets; ddd is doublet of doublet of doublets; dt is doublet of triplets; DCM is dichloromethane; methylene chloride; DIPEA is N-ethyldiisopropylamine, also known as N,N-diisopropylethylamine; DMAP is 4-dimethylaminopyridine; DMF is N,N-dimethylformamide; DMSO is dimethyl sulfoxide; DMSO-d6 is deuterated dimethyl sulfoxide; ee is the enantiomeric excess; ESI is electrospray ionization; Et2O is diethyl ether; EtOAc is ethyl acetate; EtOH is ethanol; Et3N is triethylamine; g is gram; HPLC is high performance liquid chromatography; hr is hour; L is liter; LCMS is liquid chromatography mass spectrometry; m is multiplet; M is mole; m-CPBA is 3-chloroperbenzoic acid; MeOD_d4 is deuterated methanol; MeOH is methanol; 2-MeTHF is 2-methyltetrahydrofuran; mg is milligram; MHz is megahertz; min is minute; mL is milliliter; mmol is millimole; mol is mole; MOM is methoxymethyl ether group; MS(m / z) is mass spectrum peak; NMR is nuclear magnetic resonance; Pd / C is palladium on carbon; Pd(dppf)Cl2 is [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II); pH is hydrogen ion exponent; ppm is parts per million; psi is pound per square inch; q is quartet; rpm is revolutions per minute; rt is room temperature; RT is retention time; RuPhos Pd G3 is (2-dicyclohexylphosphino-2’,6’-diisopropoxy-1,1’-biphenyl)[2-(2’-amino-1,1’-biphenyl)]palladium(II) methanesulfonate (CAS Number: 1445085-77-7); s is singlet; SEMCl is 2-(trimethylsilyl)ethoxymethyl chloride; SEM is 2-(trimethylsilyl)ethoxymethyl; SFC is supercritical fluid chromatography; t is triplet; TBAF is tert-butylammonium fluoride; TFA is trifluoroacetic acid; THF is tetrahydrofuran; TLC is thin layer chromatography; TMSCN is trimethylsilyl cyanide; TsCl is p-toluenesulfonyl chloride; μL is microliter; μmol is micromole.

[0291] In the schemes described below, it is intended to provide an overview of the methods used in the preparation of the compounds of the present invention. Some of the compounds of the present invention contain a single chiral center. In the following schemes, the general methods for preparing the compounds are shown in either racemic or enantiomerically enriched form. It will be apparent to those skilled in the art that all synthetic transformations can be carried out in exactly the same manner regardless of whether the substance is enantiomerically enriched or racemic. Further, the resolution to the desired optically active substance can be carried out at any desired point in the sequence using well-known methods such as those described in this specification and in the chemical literature.

[0292] General Method: Unless otherwise stated, the variables in Schemes I to III have the same meanings as those defined in this specification.

[0293]

Chemical formula

[0294] As exemplified in Scheme I, when 4,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidine (CAS#: 2454396-80-4) is treated with an amine in a suitable solvent (such as DCM) in the presence of an effective base (such as DIPEA), an adduct can be obtained by the SnAr reaction at the 4-chloro position. In the Suzuki reaction at the 7-chloro position, a naphthol group is incorporated in a suitable solvent such as dioxane / water using a palladium catalyst (such as CataCXium A Pd G3) and a base (such as K2CO3 or K3PO4). Oxidation of the 2-thiomethyl group to a sulfone can be carried out in a solvent (such as DCM) using an oxidizing agent (such as mCPBA). The resulting sulfone group can be replaced with an alcohol nucleophile (such as ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methanol, CAS# 2097518-76-6, etc.) in a suitable solvent (such as DCM) using a suitable base (such as LHMDS). In some cases, the penultimate intermediate may contain a protecting group, which can be removed using conditions known in the art by additional steps in the synthetic sequence (March’s Advanced Organic Chemistry: Reactions, Mechanisms, and Structure 8th Edition or Protecting Groups, 10 Georg Thieme Verlag, 1994). The compounds at each step can be purified by standard techniques such as column chromatography, crystallization, reverse-phase HPLC or SFC.

[0295]

Chemical formula

[0296] As exemplified in Scheme II, when 2,4,7-trichloro-8-fluoropyrido[4,3-d]pyrimidine (CAS# 2454396-80-4) is treated with an amine in a suitable solvent (such as DCM) in the presence of an effective base (such as DIPEA), an adduct can be obtained by the SnAr reaction at the 4-chloro position. In a second SnAr reaction at the 2-chloro position, in the presence of a base (such as DIPEA), in a suitable solvent (such as 1,4-dioxane), at a high temperature (such as 90 °C), an alcohol nucleophile (((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methanol, CAS# 2097518-76-6, etc.) is incorporated. In the Suzuki reaction at the 7-chloro position, a palladium catalyst (such as Pd(OAc)2 / dppf) and a base (such as NaOH) are used to incorporate a naphthol group in a suitable solvent such as CH3CN / water.

[0297]

Chemical formula

[0298] As exemplified in Scheme III, when 4,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidine (CAS# 2454491-14-4) is treated with a placeholder amine (such as 2-(((tert-butyldimethylsilyl)oxy)methyl)piperidine) in a suitable solvent (such as DCM) in the presence of an effective base (such as DIPEA), an adduct can be obtained by the SnAr reaction at the 4-chloro position. In the Suzuki reaction at the 7-chloro position, a palladium catalyst (such as CataCXium Pd G3) and a base (such as K3PO4) are used to incorporate a naphthol group in a suitable solvent (such as THF / water). Oxidation of the thiomethyl group to a sulfone can be achieved using an oxidizing agent (such as buffered oxone) in a solvent (such as acetone / water). In the second SnAr reaction at the 2-sulfonyl position, an alcohol nucleophile (such as ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methanol) is incorporated at a high temperature (such as 80 °C) in a suitable solvent (such as CH3CN) in the presence of a base (such as LiOTMS). The placeholder amine can be removed at a high temperature (such as 60 °C) using a base (such as NaOH) in the presence of a fluoride source (such as TBAF). Subsequently, the desired C4 amine is added in a suitable solvent (such as 2-MeTHF) in the presence of 2-chloro-1-methylpyridinium iodide and an effective base (such as DIPEA) to obtain an adduct by the SnAr reaction at the 4-oxo position.

[0299]

Chemical Structure

[0300] As illustrated in Scheme IV, when 4,5,7-trichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidine is treated with an amino alcohol (cyclic or acyclic) in a suitable solvent (such as DCM) in the presence of an effective base (such as DIPEA), an adduct can be obtained by the SnAr reaction at the 4-chloro position. In the Suzuki reaction at the 7-chloro position, a naphthol or naphthyl group is incorporated in a suitable solvent such as dioxane / water using a palladium catalyst (such as RuPhos Pd G3) and a base (such as K2CO3 or K3PO4). Oxidation of the 2-thiomethyl group to a sulfone can be carried out using an oxidizing agent (such as Oxone) in a buffered aqueous solvent containing either NaHCO3 and acetone or methyl ethyl ketone. The resulting sulfone group can be replaced with an alcohol nucleophile (such as ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methanol, CAS# 2097518-76-6, etc.) in a suitable solvent (such as CH3CN) using a suitable base (such as LHMDS or LiOTMS). In some cases, the penultimate intermediate may contain a protecting group, which can be removed using conditions known in the art by additional steps in the synthetic sequence (March’s Advanced Organic Chemistry: Reactions, Mechanisms, and Structure 8th Edition or Protecting Groups, 10 Georg Thieme Verlag, 1994). The compounds at each step can be purified by standard techniques such as column chromatography, crystallization, reverse-phase HPLC, or SFC.

[0301] Variable R in Schemes I - III 1 is the same as that defined in Embodiments E1 - E31 herein.

[0302] In Schemes I - III, the variable R represents 1 to 4 substituents selected from the group consisting of -OH, halogen, CN, C1 - C3 alkyl, C1 - C3 fluoroalkyl, C1 - C3 alkoxy, and C2 - C3 alkynyl as defined in the embodiments and claims herein.

[0303] In Schemes I - III, the variable R' may be, but is not limited to, H or C1 - C5 alkyl, and two R's may together form a ring. R' is not part of the definition in the claims herein.

[0304] The amine as presented in Schemes I - III

[0305]

Chemical Formula

[0306]

Chemical Formula

[0307] The variable R in Scheme IV 1 is the same as that defined in Embodiments E32 - E60 herein.

[0308] In Scheme IV, the variable R represents 1 to 4 substituents selected from the group consisting of -OH, halogen, CN, C1 - C3 alkyl, C1 - C3 fluoroalkyl, C1 - C3 alkoxy, and C2 - C3 alkynyl as defined in the embodiments and claims herein.

[0309] The variable R’ in Scheme IV may be, but is not limited to, H or C1-C5 alkyl, and two R’s may together form a ring. R’ is not part of the definition in the claims herein.

[0310] The amino alcohol in Scheme IV is a representative generic moiety that can form the desired tetracyclic ring structure as defined in embodiments such as E32 (wherein R 2 and R 5 may together form a 4- to 8-membered heterocycloalkyl) and E39. The amino alcohol can be an acyclic moiety that can form the desired tricyclic ring structure as defined in embodiments such as E32 where R 2 and R 5 do not together form a 4- to 8-membered heterocycloalkyl.

[0311] The synthetic intermediates generally defined in the above scheme are useful for preparing the compounds of the present invention, and the synthesis of such non-commercially available intermediates is provided as a further aspect of the present invention.

[0312] Preparation Examples: Preparation Example 1: (1R,5R,6R)-3-azabicyclo[3.2.1]octan-6-ol

[0313]

Chemical Formula

[0314] Preparation Example 2-(+) and Preparation Example 2-(-): tert-Butyl tyl-(1R * ,5R * ,6R * )-6-Hydroxy-8-oxa-3-azabicyclo[3.2.1]octane-3-carboxylate and tert-butyl tyl-(1S * ,5S * ,6S * )-6-Hydroxy-8-oxa-3-azabicyclo[3.2.1]octane-3-carboxylate

[0315] [Chemical formula] A racemic mixture of (+ / -) 2a was prepared as described in U.S. Patent 2013 / 0079321. An optical mixture of (+ / -) 2a (3.80 g, 16.6 mmol) was treated with Boc2O (5.7 g, 26 mmol) and Pd(OH)2 on carbon (4 g) in EtOH (40 mL) at 50 °C under H2 30 psi overnight. The reaction mixture was filtered through a pad of celite and the filtrate was concentrated. The residue was suspended in petroleum ether (30 mL), stirred for 1 hour, and filtered. The filter cake was dried in vacuo to give a racemic mixture of (+ / -) 2b (3.4 g, 87%) as a white solid. 1 1H NMR (400 MHz, CDCl3) δ = 4.32 (ddd, J = 4.1, 6.0, 10.6 Hz, 1H), 4.14 (br s, 1H), 4.05 - 3.87 (m, 2H), 3.56 (br s, 1H), 3.08 (br s, 2H), 2.96 (br d, J = 12.9 Hz, 1H), 2.38 (ddd, J = 7.9, 10.8, 12.4 Hz, 1H), 1.36 (s, 9H), 1.21 - 1.13 (m, 1H), MS: 130 [M+H-Boc] + .

[0316] (+ / -)2b's racemic mixture was resolved as follows using chiral SFC: Using chiral SFC (Chiralpak IG SFC 5um 21×250mm column, mobile phase with a constant composition of 90 / 10 CO2 / MeOH, 120 bar, flow rate 70 mL / min), 850 mg of (+ / -)2b was separated into its enantiomers. Peak 1 = Preparation Example 2-(+): [α] D 22 +10.7 (c 0.3, MeOH), 392 mg, >99.0% ee. Peak 2 = Preparation Example 2-(-): [α] D 22 -27.1 (c 0.1, MeOH), 294 mg, 98% ee.

[0317] Preparation Example 3: (S)-2-(1-acetylpiperazin-2-yl)acetonitrile

[0318]

Chemical Structure

[0319] Preparation Example 4: (S)-2-(3-methylpyrrolidin-3-yl)acetonitrile

[0320]

Chem.

[0321] Preparation Example 5: (S)-2-(3-Methylpiperidin-3-yl)acetonitrile

[0322]

Chemical Structure

[0323] Preparation Example 6: rac-(4-Fluoropyrrolidin-3-yl)acetonitrile

[0324]

Chemical Structure

[0325] Preparation Example 7: Trimethyl[2-({[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-{[tri(propan-2-yl)silyl]ethynyl}naphthalen-2-yl]oxy}methoxy)ethyl]silane.

[0326]

Chemical Structure

[0327] Preparation Example 8: (7,8-Difluoro-3-hydroxynaphthalen-1-yl)boronic acid.

[0328] [Chem.] The compound 1-(6-bromo-2,3-difluorophenyl)ethan-1-one (8a) (7.39 g, 31.4 mmol) was dissolved in DMF (79 mL). Pd(OAc)2 (706 mg, 3.14 mmol), (tert-Bu)3P-HBF4 (1.82 g, 6.29 mmol), and LiF (4.89 mg, 189 mmol) were then added, followed by tert-butyl((1-methoxyvinyl)oxy)dimethylsilane (18.4 g, 97.5 mmol). N2 was bubbled through the mixture for 10 minutes, and the reaction was heated at 70 °C for 30 minutes. The reaction was cooled to room temperature, and H2O (50 mL) and EtOAc (100 mL) were added. The layers were separated, and the aqueous layer was extracted with another portion of EtOAc (100 mL). The combined organic extracts were washed with brine (5 × 50 mL) and concentrated. The residue was dissolved in Et2O (100 mL), heptane (200 mL) was added, and then the heptane was removed to azeotropically remove the residual DMF. The resulting residue was purified using flash chromatography eluting with a gradient of 0–100% EtOAc in heptane to afford 8b (6.24 g, 87%) as an oil. 1 H NMR (400 MHz, CDCl3) δ 7.18 (dt, J = 9.5, 8.3 Hz, 1H), 6.97 (ddd, J = 8.4, 4.4, 1.7 Hz, 1H), 3.77 (s, 2H), 3.68 (s, 3H), 2.62 (d, J = 3.8 Hz, 3H). A solution of 8b (6.24 g, 27.3 mmol) in THF (273 mL) at -78 °C was added dropwise with LHMDS (1 M in THF 49.2 mL, 49.2 mmol). The reaction turned red and was stirred for 5 minutes, then the -78 °C bath was replaced with an ice bath. After 15 minutes, based on LCMS analysis, the reaction was complete and it was quenched by the addition of 2 M HCl (100 mL). EtOAc (100 mL) was added and stirring was continued for several minutes. The layers were separated and the aqueous layer was extracted with EtOAc (3 × 50 mL). The combined organic extracts were washed with brine (50 mL) and dried over sodium sulfate to give 8c (4.91 g, 91%) as a red solid, which was used in the next step without further purification. A solution of MeOH (125 mL) was cooled to 0 °C and acetyl chloride (37.3 g, 476 mmol) was added dropwise. After a slight exotherm, the reaction reached 0 °C again. Then, a solution of 8c (4.91 g, 25.0 mmol) in MeOH (50 mL) was added. The reaction was charged to a reflux condenser and heated at 60 °C for 6.5 hours. The reaction was concentrated and purified by flash chromatography using a gradient of 0 - 50% EtOAc in heptane to give 8d (3.78 g, 72%) as an off-white solid. 1 1H NMR (400 MHz, CDCl3) δ 7.43 (ddd, J = 9.2, 4.7, 1.8 Hz, 1H), 7.29 - 7.21 (m, 1H), 6.74 - 6.69 (m, 2H), 6.63 (d, J = 22.8 Hz, 1H), 3.88 (s, 3H). A solution of 8d (3.78 g, 40.2 mmol) in DCM (90 mL) was added with Et3N (3.64 g, 36.0 mmol), N,N-Bis(trifluoromethylsulfonyl)aniline (7.71 g, 21.6 mmol) and DMAP (110 mg, 0.899 mmol). The reaction was stirred at room temperature for 8 h. Additional N,N-Bis(trifluoromethylsulfonyl)aniline (1.29 g, 3.60 mmol) was added and the reaction was stirred for an additional 11 h. The mixture was transferred to a separatory funnel and washed with 1N NaOH (50 mL). The DCM layer was dried over Na2SO4, filtered and purified by flash chromatography eluting with a gradient of 0 - 30% EtOAc in heptane to give 8e (6.7 g) as a solid in yield above theory. Since the NMR showed low purity, the material was re-purified using a 220 g Gold Isco column eluting with a gradient of 0 - 30% EtOAc in heptane to give 8e (5.18 g, 84%) as an off-white solid. 1 H NMR (400 MHz, CDCl3) δ 7.53 (ddd, J = 9.2, 4.6, 1.9 Hz, 1H), 7.39 (td, J = 9.4, 7.3 Hz, 1H), 7.20 (d, J = 2.3 Hz, 1H), 7.14 (t, J = 2.1 Hz, 1H), 3.94 (s, 3H), MS: 210 [M+H-SO2CF3] +Ethanol (56 mL) and DIPEA (7.91 mL, 45.4 mmol) were added to a flask containing 8e (5.18 g, 15.1 mmol). Tetrahydroxydiboron (2.04 g, 22.7 mmol), 1,3-bis(diphenylphosphino)propanenickel(II) chloride (410 mg, 0.757 mmol), and triphenylphosphine (397 mg, 1.51 mmol) were added. Nitrogen was bubbled through the mixture for 5 minutes. The reaction was heated to 50 °C for 17 hours. LCMS showed one new peak that was more polar but did not ionize. The mixture was diluted with EtOAc and the product was extracted with 1N NaOH (2 × 75 mL). The aqueous layer was acidified to pH = 1 with 6N HCl and extracted with EtOAc (2 × 200 mL). The EtOAc was dried over Na2SO4 and concentrated to give (7,8-difluoro-3-hydroxynaphthalen-1-yl)boronic acid, Preparation Example 8 (3.6 g, 99%) as a white solid. 1 H NMR (400 MHz, CDCl3) δ = 7.50 (ddd, J = 1.7, 4.7, 9.0 Hz, 1H), 7.42 (d, J = 2.3 Hz, 1H), 7.32 (dt, J = 7.9, 9.4 Hz, 1H), 7.14 (t, J = 2.1 Hz, 1H), 3.93 (s, 3H). The B(OH)2 proton appeared as a very broad peak between 2 - 3 ppm depending on the concentration. 19 F NMR (376 MHz, CDCl3) δ = -141.77 (d, J = 18 Hz, 1F), -144.32 (d, J = 20 Hz, 1F).

[0329] Preparation Example 9: 4,5,7-Trichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidine

[0330]

Chemical Structure

[0331] Preparation Example 10: (8aS)-5-chloro-4-fluoro-2-(methylsulfanyl)-8,8a,9,10,11,12-hexahydro-7-oxa-1,3,6,12a-tetraazabenzo[4,5]cyclohepta[1,2,3-de]naphthalene

[0332] [Chemical Structure] Suspension of Preparation Example 9 (1.25 g, 3.70 mmol) in CH3CN (24 mL), addition of DIEA (0.668 mL, 3.83 mmol). The mixture was cooled to 0 °C, and (S)-piperidin-2-ylmethanol (421 mg, 3.65 mmol) was added as a solution in THF (18 mL). As observed by LCMS, after 8 minutes, the first nitrogen-carbon bond was formed. LiOtBu (877 mg, 11.0 mmol) was added as a solution in THF (22 mL), and the mixture was warmed to 50 °C. After 4 hours at 50 °C, LCMS analysis showed conversion to Preparation Example 9. Then, the reaction mixture was diluted with 200 mL of water, and the product was extracted with DCM (50 mL × 4). The combined organic extracts were dried over Na2SO4, filtered, and evaporated to obtain Preparation Example 10 as a crude solid. Purification was achieved by flash chromatography eluting with a gradient of 0 - 10% MeOH in DCM to give 1.13 g (91%) of Preparation Example 10. 1 H NMR (chloroform-d, 400 MHz) δ 4.8 - 4.9 (m, 1H), 4.4 - 4.5 (m, 2H), 3.7 - 3.8 (m, 1H), 2.97 (dt, 1H, J = 2.5, 12.8 Hz), 2.7 - 2.7 (m, 1H), 2.6 - 2.7 (m, 2H), 2.0 - 2.1 (m, 1H), 1.7 - 1.8 (m, 3H), 1.5 - 1.7 (m, 2H), MS: 341.1 [M+H] + .

[0333] Preparation Example 11: (8aS)-4-Fluoro-5-[7-fluoro-3-(methoxymethoxy)-8-{[tri(propan-2-yl)silyl]ethynyl}naphthalen-1-yl]-2-(methylsulfanyl)-8,8a,9,10,11,12-hexahydro-7-oxa-1,3,6,12a-tetraazabenzo[4,5]cyclohepta[1,2,3-de]naphthalene

[0334]

Chem.

[0335] Preparation Example 12: (8aS)-4-Fluoro-5-[7-fluoro-3-(methoxymethoxy)-8-{[tri(propan-2-yl)silyl]ethynyl}naphthalen-1-yl]-2-{[(2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl]methoxy}-8,8a,9,10,11,12-hexahydro-7-oxa-1,3,6,12a-tetraazabenzo[4,5]cyclohepta[1,2,3-de]naphthalene

[0336]

Chemical formula

[0337] Preparation Example 13: {[2,3-Difluoro-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-6-{[2-(trimethylsilyl)ethoxy]methoxy}naphthalen-1-yl]ethynyl}tri(propan-2-yl)silane

[0338]

Chemical formula

Examples

[0339] The following examples are described to better understand the present invention. These examples are for illustrative purposes only and should not be construed as limiting the scope of the present invention in any way.

[0340] (Example 1) {(3S)-1-[7-(8-Ethynyl-3-hydroxynaphthalen-1-yl)-8-fluoro-2-{[(2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl]methoxy}pyrido[4,3-d]pyrimidin-4-yl]piperidin-3-yl}acetonitrile.

[0341]

Chemical Formula

[0342] To a solution of CAS# 2454396-80-4 (215 mg, 0.814 mmol) and CAS# 1693757-39-9 (131 mg, 0.819 mmol) in DCM (10 mL), DIPEA (316 mg, 2.44 mmol) was added at -40 °C under argon. The mixture was stirred at 25 °C for 2 h. The mixture was warmed to room temperature and diluted with water (30 mL). The aqueous layer was extracted with DCM (3 × 30 mL). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by flash chromatography using a gradient of 0 - 60% EtOAc in petroleum ether to give 1A (160 mg, 56%) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.88 (s, 1H), 4.48 (d, J = 11.6 Hz, 1H), 4.38 (d, J = 13.4 Hz, 1H), 3.38 (t, J = 10.9 Hz, 1H), 3.27 - 3.20 (m, 1H), 2.63 (dd, J = 6.7, 4.0 Hz, 2H), 2.59 - 2.52 (m, 3H), 2.14 - 2.05 (m, 1H), 1.94 (d, J = 9.8 Hz, 1H), 1.87 - 1.79 (m, 1H), 1.71 - 1.62 (m, 1H), 1.49 - 1.40 (m, 1H), MS: 352 [M+H] + To a solution of 1A (160 mg, 0.455 mmol) and Preparation Example 7 (343 mg, 0.591 mmol) in dioxane (10 mL) and H2O (1 mL), CataCXium A Pd G3 (33.1 mg, 0.0455 mmol) and K2CO3 (189 mg, 1.36 mmol) were added in-tube under N2. The reaction was stirred at 80 °C for 6 h. The mixture was concentrated under reduced pressure and the residue was purified by flash chromatography using a gradient of 0 - 50% EtOAc in petroleum ether to give 1B (207 mg, 59%) as a yellow solid. 11H NMR (400 MHz, DMSO) δ 9.10 (d, J = 4.6 Hz, 1H), 8.04 (d, J = 8.3 Hz, 1H), 7.72 (s, 1H), 7.69 - 7.63 (m, 1H), 7.58 (t, J = 7.7 Hz, 1H), 7.33 (d, J = 2.3 Hz, 1H), 5.46 (s, 2H), 4.57 (d, J = 11.8 Hz, 1H), 4.48 (s, 1H), 3.88 - 3.76 (m, 2H), 3.20 (d, J = 10.8 Hz, 1H), 2.71 (d, J = 6.8 Hz, 2H), 2.60 (d, J = 2.2 Hz, 3H), 2.10 - 1.92 (m, 3H), 1.51 (d, J = 11.7 Hz, 1H), 1.28 (s, 1H), 1.00 - 0.94 (m, 2H), 0.93 - 0.80 (m, 18H), 0.53 (dq, J = 14.4, 7.1 Hz, 3H), 0.05 (s, 1H), -0.00 (s, 9H), MS: 770 [M+H] + .A solution of 1B (207 mg, 0.269 mmol) in DCM (10 mL) was added to mCBPA (69.6 mg, 0.403 mmol). The reaction mixture was stirred at 20 °C for 1 hour. The reaction mixture was diluted with saturated aqueous Na2SO3 (20 mL) and extracted with DCM (3 × 20 mL). The combined organic layers were washed with saturated NaHCO3 (100 mL) and brine (100 mL), and dried over anhydrous Na2SO4. After concentration, crude 1C (211 mg, 99%) was obtained as a yellow solid. 1 1H NMR (400 MHz, DMSO) δ 9.31 (s, 1H), 8.07 (d, J = 8.2 Hz, 1H), 7.76 (s, 1H), 7.67 (s, 1H), 7.60 (t, J = 8.0 Hz, 1H), 7.36 (s, 1H), 5.47 (s, 2H), 4.67 (s, 2H), 3.81 (t, J = 7.8 Hz, 2H), 2.98 (dd, J = 11.1, 6.6 Hz, 3H), 2.76 (s, 2H), 2.04 (d, J = 7.8 Hz, 2H), 1.28 (s, 3H), 0.96 (t, J = 8.2 Hz, 2H), 0.86 (dd, J = 12.3, 5.1 Hz, 18H), 0.54 (dt, J = 14.9, 7.4 Hz, 3H), 0.05 (s, 3H), -0.00 (s, 9H), MS: 786 [M+H] + . A solution of 1C (211 mg, 0.263 mmol) and CAS# 2097518-76-6 (50.3 mg, 0.316 mmol) in anhydrous DCM (10 mL) was added dropwise with LHMDS (1M in THF 0.289 mL, 0.289 mmol) at 0 °C under N2. The reaction was stirred at 0 °C for 1 hour. The reaction mixture was diluted with H2O (15 mL) and extracted with DCM (3 × 10 mL). The combined organic layers were washed with brine (100 mL) and dried over anhydrous Na2SO4. After concentration under reduced pressure, the obtained residue was purified by flash chromatography eluting with a gradient of 0 - 10% MeOH in DCM to give 1D (161 mg, 69%) as a yellow solid. 1 H NMR (400 MHz, DMSO) δ 9.09 (d, J = 3.8 Hz, 1H), 8.04 (d, J = 8.2 Hz, 1H), 7.72 (d, J = 2.5 Hz, 1H), 7.69 - 7.63 (m, 1H), 7.62 - 7.54 (m, 1H), 7.32 (d, J = 1.9 Hz, 1H), 5.46 (s, 2H), 5.27 (s, 1H), 4.48 (dd, J = 42.3, 11.5 Hz, 3H), 4.18 (dd, J = 23.5, 10.5 Hz, 1H), 4.09 - 3.97 (m, 1H), 3.90 - 3.67 (m, 2H), 3.19 (dd, J = 25.0, 11.6 Hz, 3H), 3.07 (s, 1H), 2.89 (s, 1H), 2.77 - 2.67 (m, 2H), 2.11 - 2.01 (m, 2H), 1.96 (d, J = 13.2 Hz, 2H), 1.80 (d, J = 11.7 Hz, 2H), 1.52 (d, J = 13.4 Hz, 2H), 0.97 (dd, J = 18.9, 10.0 Hz, 3H), 0.91 - 0.82 (m, 18H), 0.54 (tt, J = 15.0, 6.0 Hz, 3H), 0.11 - 0.02 (m, 3H), 0.02 - -0.04 (m, 9H), MS: 881 [M+H] + . A solution of 1D (160 mg, 0.182 mmol) in DMF (5 mL) was added with CsF (276 mg, 1.82 mmol). Subsequently, the reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was diluted with H2O (30 mL) and extracted with EtOAc (3 × 20 mL). The combined organic layers were washed with brine (100 mL) and dried over anhydrous Na2SO4. After concentration under reduced pressure, the crude terminal alkyne (132 mg, 99%) was obtained as a yellow solid and used for the next step without further purification. MS: 725 [M+H] + . HCl (4 M 0.20 mL, 0.80 mmol in dioxane) was added to a solution of the crude terminal alkyne (120 mg, 0.166 mmol) in DCM (10 mL). The reaction mixture was stirred at 25 °C for 0.5 hour. The reaction mixture was concentrated, and the crude product was purified by preparative HPLC (column: Xbridge 5μm C18 150×19mm; mobile phase: CH3CN - water (0.1% formic acid); gradient: 23% - 100%; flow rate: 20 mL / min) to obtain {(3S)-1-[7-(8-ethynyl-3-hydroxynaphthalen-1-yl)-8-fluoro-2-{[(2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl]methoxy}pyrido[4,3-d]pyrimidin-4-yl]piperidin-3-yl}acetonitrile, Example 1 (25.2 mg, 24%) as a yellow solid.1 1H NMR (400 MHz, DMSO-d6) δ 9.00 (d, J = 10.0 Hz, 1H), 7.88 (d, J = 8.0 Hz, 1H), 7.50 - 7.39 (m, 2H), 7.34 (d, J = 2.4 Hz, 1H), 7.14 (d, J = 2.3 Hz, 1H), 5.48 - 5.15 (m, 1H), 4.45 (dd, J = 25.6, 13.0 Hz, 3H), 4.25 - 4.10 (m, 1H), 4.10 - 4.00 (m, 1H), 3.62 (d, J = 3.7 Hz, 1H), 3.20 - 2.97 (m, 5H), 2.83 (d, J = 6.6 Hz, 1H), 2.66 (d, J = 7.5 Hz, 2H), 2.13 (d, J = 4.7 Hz, 2H), 2.05 (s, 1H), 2.01 (s, 2H), 1.92 - 1.71 (m, 4H), 1.47 (s, 1H), MS: 595 [M+H] + .

[0343] Examples 2 - 9 reported in Table 1 were prepared according to Scheme I / General Method A with minor modifications that would be recognized by those skilled in the art.

[0344] (Example 10) (1R,5R,6R)-3-[7-(8-Ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-2-{[(2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl]methoxy}pyrido[4,3-d]pyrimidin-4-yl]-3-azabicyclo[3.2.1]octan-6-ol.

[0345]

Chemical formula

[0346] A solution of CAS# 2454396-80-4 (913 mg, 3.61 mmol) and Preparation Example 1 (591 mg, 3.61 mmol) was dissolved in DCM (72 mL). The solution was cooled to -78 °C and DIPEA (1.57 mL, 9.04 mmol) was added. The reaction was stirred at -78 °C for 1 hour and the cold bath was removed. The reaction was warmed to room temperature over 1 hour. The solvent was removed and the residue was purified by flash chromatography eluting with a gradient of 0 - 100% EtOAc in heptane. After concentration of the pure fractions, 10A (1.09 g, 88%) was obtained as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.17 - 1.24 (m, 1 H) 1.69 (s, 1 H) 1.79 (s, 1 H) 2.05 - 2.15 (m, 1 H) 2.16 - 2.22 (m, 1 H) 2.33 - 2.38 (m, 1 H) 3.45 (br d, J = 12.6 Hz, 1 H) 3.74 (br d, J = 12.5 Hz, 1 H) 4.15 - 4.22 (m, 1 H) 4.46 (br s, 1 H) 4.58 (br d, J = 13.3 Hz, 1 H) 4.66 - 4.73 (m, 1 H) 9.21 (s, 1 H), MS: 343, 345 [M+H] + A solution of 10A (1.22 g, 3.55 mmol) in dioxane (20 mL) was treated with DIPEA (1.24 mL, 7.11 mmol) and ((2R,7AS)-2-fluorohexahydro-1H-pyrrolidin-7A-yl)methanol (1.13 g, 7.11 mmol) and the mixture was heated to 90 °C for 48 hours. The reaction was diluted with EtOAc (100 mL) and washed with water and brine. The organic extract was dried over sodium sulfate and concentrated to give an orange oil. The crude orange oil was purified by SFC to give 10B (890 mg, 54%). 11H NMR (400 MHz, DMSO-d6) δ ppm 9.08 (s, 1 H) 5.16 - 5.41 (m, 1 H) 4.62 - 4.75 (m, 2 H) 4.47 (br d, J = 12.1 Hz, 1 H) 4.08 - 4.19 (m, 2 H) 4.00 (d, J = 10.3 Hz, 1 H) 3.63 - 3.74 (m, 1 H) 3.33 (s, 1 H) 2.98 - 3.14 (m, 3 H) 2.73 - 2.91 (m, 1 H) 2.31 (br s, 1 H) 1.93 - 2.20 (m, 5 H) 1.72 - 1.91 (m, 4 H) 1.57 - 1.69 (m, 1 H) 1.11 - 1.26 (m, 1 H), MS 466 [M+H] + . To a mixture of 10B (100 mg, 0.215 mmol) and CAS# 2621932-37-2 (132 mg, 0.258 mmol) in THF (2.5 mL) were added K3PO4 (150 mg, 0.708 mmol) and water (0.25 mL). Nitrogen was bubbled through the solution for 10 minutes, and CataCXium A Pd G3 (15.6 mg, 0.0215 mmol) was added. The vial was sealed and heated at 60 °C for 2 hours. LCMS showed a clean reaction and a conversion rate of approximately 50% to the Suzuki product. Heating was continued at 60 °C for an additional 18 hours. The mixture was concentrated under vacuum, and the resulting residue was purified by flash chromatography eluting with a gradient of 0 - 20% iPrOH in DCM. After concentrating the pure fractions, 10C (130 mg, 74%) was obtained as a yellow powder. 1 1H NMR (400 MHz, DMSO-d6) δ = 9.62, 9.26 (2s, 1H, each for the rotational enantiomer and the minor rotational enantiomer), 8.10 (dd, J = 5.9, 9.3 Hz, 1H), 7.73 (d, J = 2.4 Hz, 1H), 7.56 (t, J = 8.9 Hz, 1H), 7.39 - 7.31 (m, 1H), 5.40 - 5.34 (m, 2H), 5.23 (br s, 1H), 4.97 - 4.84 (m, 1H), 4.68, 4.50 (2d, J = 3.4 Hz, br d, J = 11.5 Hz, 1H, each for the rotational main isomer and the minor rotational main isomer), 4.32 - 4.05 (m, 3H), 4.01 - 3.91 (m, 1H), 3.82 - 3.73 and 3.60 - 3.53 (2m, 1H, each for the rotational main isomer and the minor rotational main isomer), 3.44 (s, 2H), 3.16 - 2.99 (m, 2H), 2.91 - 2.79 (m, 1H), 2.52 - 2.35 (m, 4H are hidden in the DMSO peak), 2.40 - 2.30 (m, 1H), 2.26 - 1.96 (m, 4H), 1.91 - 1.61 (m, 4H), 1.36 - 1.22 (m, 1H), 0.87 - 0.78 (m, 18H), 0.56 - 0.42 (m, 3H); MS: 816 [M+H] +A solution of 10C (126 mg, 0.154 mmol) in CH3CN (5 mL) was treated with CsF (235 mg, 1.55 mmol). The reaction was stirred at room temperature for 4 h. The mixture was neutralized with acetic acid (22 mL, 0.386 mmol) and concentrated in vacuo. The residue was taken up in EtOAc and the salts were removed by filtration. The filtrate was concentrated to give the terminal alkyne as a pink-colored powder (102 mg) which was carried on to the next step without further purification. A suspension of the terminal alkyne (102 mg, 0.154 mmol) from the previous step in CH3CN (2 mL) was cooled in an ice bath. 4N HCl (2 mL of 4N in dioxane, 8.0 mmol) was added. The reaction was stirred at 0 °C for 1 h. The solvent was evaporated to give an orange solid. This residue was dissolved in MeOH (1 mL) and purified by preparative HPLC using acetic acid as an additive to the CH3CN / water eluent. The pure fractions were combined and concentrated to 15 mL of water. The aqueous solution was frozen at -78 °C and lyophilized overnight to give (1R,5R,6R)-3-[7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-2-{[(2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl]methoxy}pyrido[4,3-d]pyrimidin-4-yl]-3-azabicyclo[3.2.1]octan-6-ol, Example 10 (104 mg, 64%) as a pale orange powder. 1 H NMR (400 MHz, methanol-d4) δ = 9.28 - 9.04 (m, 1H), 7.90 - 7.81 (m, 1H), 7.35 (d, J = 2.4 Hz, 1H), 7.34 - 7.28 (m, 1H), 7.27 - 7.19 (m, 1H), 5.49 - 5.30 (m, 1H), 5.21 (br d, J = 11.9 Hz, 1H), 4.67 (br d, J = 12.1 Hz, 1H), 4.50 - 4.42 (m, 1H), 4.38 - 4.26 (m, 2H), 3.98 - 3.72 (m, 1H), 3.58 - 3.34 (m, 5H), 3.21 - 3.11 (m, 1H), 2.51 - 2.05 (m, 9H), 2.03 - 1.89 (m, 5H), 1.86 - 1.75 (m, 1H), 1.46 - 1.36 (m, 1H), HRMS: 616.25385 [M+H] + .

[0347] Examples 11 - 17 reported in Table 1 were prepared according to General Method B / Example 10 with minor modifications that would be recognized by those skilled in the art.

[0348] (Example 18) (1R * ,5R * ,6R * )-3-[7-(8-Ethynyl-3-hydroxynaphthalen-1-yl)-8-fluoro-2-{[(2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl]methoxy}pyrido[4,3-d]pyrimidin-4-yl]-8-oxa-3-azabicyclo[3.2.1]octan-6-ol.

[0349]

Chemical Structure

[0350] Example 18 was prepared according to the above scheme, which represents General Method C.

[0351] Compound CAS# 2454491-14-4 (3.66 g, 13.8 mmol) was combined with CAS# 135938-63-5 (3.18 g, 13.9 mmol) in DCM (69 mL). The resulting solution was cooled to -40 °C and DIPEA (7.0 mL, 42 mmol) was added. The mixture was gradually warmed from -40 °C to room temperature over 3.5 hours. The mixture was partitioned between water and DCM, and the DCM layer was washed with water (3×). The organic layer was dried over sodium sulfate, and fresh CAS# 135938-63-5 (1.6 g, 6.9 mmol) and further fresh DIPEA (7.0 mL, 42 mmol) were added and the reaction conditions were applied again. After stirring for 1 hour, a third portion of CAS# 135938-63-5 (1.6 g, 6.9 mmol) was added and the reaction was stirred at room temperature for 18 hours. The reaction mixture was partitioned between water and DCM, and the DCM layer was washed with water (3×). The organic layer was dried over sodium sulfate and evaporated. Purification by flash chromatography using a gradient of 0 - 100% EtOAc in heptane gave 18A (4.26 g, 67%) as a glassy solid. 1 H NMR (400 MHz, CDCl3) δ 8.94 (s, 1H), 4.84 (s, 1H), 4.46 (d, J = 13.5 Hz, 1H), 4.12 (dd, J = 10.5, 8.6 Hz, 1H), 3.76 (dd, J = 10.5, 5.7 Hz, 1H), 3.34 (t, J = 12.3 Hz, 1H), 2.60 (s, 3H), 1.88 - 1.64 (m, 9H), 0.79 (s, 8H), 0.02 (d, J = 2.3 Hz, 6H), 19 F NMR (376 MHz, CDCl3) δ -134.84, MS: 457.1 [M+H] +0.18 A (197 mg, 0.430 mmol) was combined with Preparation Example 7 (500 mg, 0.861 mmol) in THF (4.3 mL) and aqueous K3PO4 solution (1.5 M 1.43 mL, 2.15 mmol). The mixture was purged with nitrogen for 3 minutes. Then, CataCXium A Pd G3 (31.3 mg, 0.0430 mmol) was added and the mixture was further purged with nitrogen for 3 minutes. The reaction was heated at 70 °C for 4.5 hours and the formation of the product was observed by TLC. The mixture was then evaporated directly onto celite. The celite was packed into an Isco cartridge and the product was purified by flash chromatography eluting with a gradient of 0 - 100% EtOAc in heptane. The fractions were analyzed using a non-polar / high mass LCMS method which picked up the target mass as a late eluting peak. The pure fractions were concentrated to give Compound 18B (367 mg, 49%) as a glassy orange solid. 1 H NMR (400 MHz, CDCl3) δ 9.11 (d, J = 20.0 Hz, 1H), 7.81 (dt, J = 8.3, 1.5 Hz, 1H), 7.68 (dd, J = 7.2, 1.2 Hz, 1H), 7.54 (t, J = 3.0 Hz, 1H), 7.42 - 7.38 (m, 1H), 7.30 (dd, J = 4.3, 2.6 Hz, 1H), 5.37 (d, J = 7.0 Hz, 1H), 5.35 - 5.31 (m, 1H), 4.87 (s, 1H), 4.38 (dd, J = 38.0, 13.8 Hz, 1H), 4.05 - 3.96 (m, 1H), 3.84 - 3.74 (m, 3H), 3.51 - 3.32 (m, 1H), 2.61 (d, J = 1.3 Hz, 3H), 1.27 (q, J = 2.2 Hz, 4H), 1.19 - 1.16 (m, 4H), 1.05 (d, J = 1.1 Hz, 3H), 1.00 - 0.95 (m, 3H), 0.90 (d, J = 1.6 Hz, 9H), 0.88 (t, J = 1.8 Hz, 9H), 0.77 (s, 6H), 0.58 - 0.49 (m, 3H), -0.00 (t, J = 1.2 Hz, 9H), -0.03 (d, J = 2.5 Hz, 3H), MS: 875.4 [M+H] + . Compound 18B (367 mg, 0.419 mmol) was dissolved in a mixture of acetone (20 mL) and saturated aqueous sodium bicarbonate solution (10 mL). Oxone (322 mg, 0.524 mmol) was added and the mixture was stirred at room temperature for 45 minutes. When the pH was examined with pH paper, it was observed to be between 7 and 8. The reaction was quenched by the addition of saturated aqueous sodium sulfite solution (20 mL) and the reaction mixture was stirred for 5 minutes. The reaction mixture was extracted using 50 / 50 heptane:EtOAc (1×40 mL), the organic extract was washed with brine (1×25 mL) and dried over sodium sulfate. After filtration and concentration, 18C (341 mg, 90%) was obtained. Note: The masses of both the sulfoxide and the sulfone were observed under a single broad peak. MS: Sulfoxide: 891.4, Sulfone: 907.4 [M+H] + . This material was taken on to the next step without further purification. Compound 18C (341 mg, 0.376 mmol) was combined with CAS# 2097518-76-6 (89.7 mg, 0.564 mmol) and lithium trimethylsilanolate (108 mg, 1.13 mmol) in CH3CN (4.18 mL). The mixture was heated at 80 °C for 30 minutes. After cooling, the solid was removed by filtration. The filtrate was diluted with water (40 mL) and extracted using a solvent mixture of 10% EtOAc in heptane (3×10 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated. The residue was purified by flash chromatography by dissolving in 5 mL of heptane and loading directly onto a silica column. Gradient elution was carried out using 0 - 100% EtOAc in heptane to give 18D (236 mg, 64%). MS: 986.5 [M+H] +Compound 18D (236 mg, 0.239 mmol) was dissolved in THF (2.39 mL), and an aqueous NaOH solution (1 M 1.20 mL, 1.20 mmol) was added, followed by TBAF (1 M 1.20 mL in THF, 1.20 mmol). The mixture was heated to 60 °C with stirring. After 2.3 hours, the reaction mixture was cooled to room temperature and diluted with EtOAc (30 mL). The organic layer was washed with brine (6 × 25 mL) to remove excess TBAF. The organic layer was dried over sodium sulfate, filtered, and evaporated. The residue was purified by flash chromatography eluting with a gradient of 0 - 100% EtOAc in heptane (containing 10% by volume of 7N ammonia in methanol). The pure fractions were collected to give (50 mg, 27%), and the column was further flashed with 75% methanol in DCM to elute the remaining target substance remaining on the column to give an additional 100 mg of product. The total yield of the C4 deprotection step was 150 mg, 81%, MS: 775.3 [M+H] + Half of this material was taken directly into the amine coupling step. The material from the TBAF / NaOH treatment (50 mg, 0.065 mmol) was combined with CMPI (26.4 mg, 0.103 mmol) and DIPEA (57.4 μL, 0.323 mmol) in DCM (1.0 mL). The mixture was stirred at room temperature for 1 hour. Then, Preparation Example 2-(-) (17.1 mg, 0.103 mmol) was added, and the mixture was stirred at room temperature for 3 hours. The reaction mixture was diluted with EtOAc (30 mL), and the organic layer was washed with brine (3 × 10 mL). The organic layer was dried over sodium sulfate, filtered, and evaporated to give 18E (52.0 mg, 91%). MS 886.4 [M+H] +18E (52 mg, 0.059 mmol) was dissolved in CH3CN (1.0 mL), and CsF (89.1 mg, 0.587 mmol) was added. The reaction mixture was stirred at room temperature for 6 hours to complete the removal of the TiPS group. Then, HCl (4.0 M in dioxane 0.293 mL, 1.17 mmol) was added. The reaction was stirred for 8 hours. The reaction mixture was diluted with EtOAc (30 mL), and the organic layer was washed with brine and a 50 / 50 solution of 1 M aqueous NaOH (3 × 20 mL). The organic layer was dried over sodium sulfate, filtered, and evaporated to obtain 46 mg of crude Example 18. Purification by reverse-phase HPLC gave (1R * ,5R * ,6R * )-3-[7-(8-ethynyl-3-hydroxynaphthalen-1-yl)-8-fluoro-2-{[(2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl]methoxy}pyrido[4,3-d]pyrimidin-4-yl]-8-oxa-3-azabicyclo[3.2.1]octan-6-ol, Example 18. 1 1H NMR (400 MHz, DMSO) δ 9.07 and 9.31 (2 s, 1H, each for the rotamer and the minor rotamer), 7.87 (dt, J = 8.0, 2.3 Hz, 1H), 7.46 - 7.39 (m, 2H), 7.34 (t, J = 2.5 Hz, 1H), 7.17 and 7.11 (2 d, J = 2.6 Hz, 1H, each for the rotamer and the minor rotamer), 5.26 (d, J = 54.3 Hz, 1H), 4.42 (d, J = 7.3 Hz, 1H), 4.33 - 4.19 (m, 2H), 4.16 - 3.98 (m, 4H), 3.44 (d, J = 11.9 Hz, 1H), 3.07 (d, J = 9.5 Hz, 2H), 2.99 (s, 1H), 2.81 (t, J = 7.9 Hz, 1H), 2.34 - 2.25 (m, 1H), 2.11 - 2.02 (m, 2H), 1.77 (d, J = 8.2 Hz, 8H),19 19F NMR (376 MHz, DMSO) δ -141.48, -172.18 (reported for major rotamer only), MS: 600.2 [M+H] + .

[0352] Examples 19 - 22 reported in Table 1 were prepared according to General Method C / Example 18 with minor modifications that would be recognized by one of ordinary skill in the art.

[0353] Additional compounds of the invention were prepared by modifying the methods exemplified herein. Unless otherwise indicated, all compounds having chiral centers were prepared and / or isolated as a single enantiomer having a known relative stereochemistry. Compounds labeled "absolute stereochemistry unknown" were typically prepared from racemic intermediates and resolved into single enantiomers by appropriate chiral preparative SFC methods prior to characterization and testing.

[0354] Examples 1 - 22 and their corresponding characterization data are all presented in Table 1 below.

[0355]

Table 1 - 1

[0356]

Table 1 - 2

[0357]

Table 1 - 3

[0358]

Table 1 - 4

[0359]

Table 1 - 5

[0360] [Table 1-6]

[0361] [Table 1-7]

[0362] [Table 1-8]

[0363] [Table 1-9]

[0364] [Table 1-10]

[0365] (Example 23) 5-Ethynyl-6-fluoro-4-[(8aS)-4-fluoro-2-{[(2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl]methoxy}-8,8a,9,10,11,12-hexahydro-7-oxa-1,3,6,12a-tetraazabenzo[4,5]cyclohepta[1,2,3-de]naphthalen-5-yl]naphthalen-2-ol

[0366] [Chemical Structure] Modulation Example 12 (910 mg, 1.2 mmol) was dissolved in CH3CN (12 mL), and CsF (1.1 g, 7.2 mmol) was added. The reaction mixture was stirred at 35 °C for 16 hours, and the reaction mixture was partitioned between water (15 mL) and EtOAc (30 mL). The aqueous layer was extracted with EtOAc (20 mL × 3), and the combined organic extracts were dried over Na2SO4, filtered, and concentrated. The resulting solid was purified by SFC using a ZymorSPHER HADP 150×21.2 column and a gradient of 15 - 40% MeOH in CO2, 100 mL / min, 110 bar to obtain 493 mg (68%) of Example 23 as a yellow solid. 1 By 1H NMR, 29 / 30 protons were observed (exchangeable phenols were not observed). 1 1H NMR (400 MHz, DMSO) δ 7.89 (dd, J = 8.8, 6.4 Hz, 1H), 7.40 (t, J = 9.6 Hz, 1H), 7.30 (d, J = 2.6 Hz, 1H), 7.13 (dd, J = 14.8, 2.5 Hz, 1H), 5.28 (d, J = 52.9 Hz, 1H), 5.20 - 5.06 (m, 1H), 4.53 - 4.36 (m, 2H), 4.12 (dd, J = 10.4, 3.2 Hz, 1H), 4.05 - 3.97 (m, 2H), 3.92 (dd, J = 9.5, 5.1 Hz, 1H), 3.13 - 3.06 (m, 2H), 3.01 (d, J = 9.2 Hz, 2H), 2.83 (q, J = 8.6 Hz, 1H), 2.20 - 2.09 (m, 1H), 2.03 (dd, J = 20.6, 3.4 Hz, 2H), 1.92 - 1.47 (m, 9H), 19 19F NMR (377 MHz, DMSO) δ -111.58, -145.15, -172.11.

[0367] The steps and reaction conditions outlined above for Example 23 are hereby defined as General Method D.

[0368] Examples 24 to 31 reported in Table 1A were prepared according to General Method D / Example 23 with minor modifications that would be recognized by those skilled in the art.

[0369] [Table 2-1]

[0370] [Table 2-2]

[0371] [Table 2-3]

[0372] [Table 2-4]

[0373] (Example 32) 5-Ethynyl-6,7-difluoro-4-[(8aS)-4-fluoro-2-{[(2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl]methoxy}-8,8a,9,10,11,12-hexahydro-7-oxa-1,3,6,12a-tetraazabenzo[4,5]cyclohepta[1,2,3-de]naphthalen-5-yl]naphthalen-2-ol

[0374] [Chemical Structure] Using General Method D / Example 23 with minor modifications that would be recognized by those skilled in the art, Boronic Acid Ester Preparation Example 13 was coupled with Preparation Example 10 to obtain Example 32 in 5 steps. The final SEM deprotection step for this is provided below.

[0375] The SEM-protected intermediate resulting in Example 32 (90 mg, 0.12 mmol) was dissolved in DCM (5 mL). To the DCM solution, HCl in dioxane (4 M 0.45 mL, 1.8 mmol) was added at 15 °C. The reaction mixture was stirred at 15 °C under N2 for 30 minutes. LCMS analysis indicated that the starting material was almost consumed. The mixture was concentrated under vacuum to give the crude product, which was purified using preparative HPLC (Waters MS triggered Prep-LC equipped with an SQD2 detector; column: Welch 10m C18 250×21.2 mm; flow rate: 25 mL / min; wavelength: 214 nm; 50% - 70% ACN in H20 (0.1% NH3)) to afford Example 32 (24 mg, 30%) as a yellow solid. 1 1H NMR (methanol-d4, 400 MHz) δ 7.6 - 7.7 (m, 1H), 7.25 (d, 1H, J = 2.6 Hz), 7.15 (dd, 1H, J = 2.5, 16.7 Hz), 5.2 - 5.4 (m, 2H), 4.4 - 4.6 (m, 2H), 4.2 - 4.4 (m, 1H), 4.1 - 4.2 (m, 1H), 3.9 - 4.0 (m, 1H), 3.68 (d, 1H, J = 9.8 Hz), 3.1 - 3.3 (m, 3H), 3.0 - 3.2 (m, 1H), 3.0 - 3.0 (m, 1H), 2.3 - 2.4 (m, 1H), 2.2 - 2.4 (m, 1H), 2.1 - 2.2 (m, 1H), 1.7 - 2.1 (m, 8H), 19 19F NMR (methanol-d4, 376 MHz) δ - 135.94 - - 136.00 (m, 1F), - 139.86 - - 139.91 (m, 1F), - 145.47 - - 145.64 (m, 1F), - 173.61 - - 173.67 (m, 1F), MS: 620 [M+H] + .

[0376] (Example 33) 5-Ethynyl-6,7-difluoro-4-[(8aS)-4-fluoro-2-{[(2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl]methoxy}-8,8a,9,10,11,12-hexahydro-7-oxa-1,3,6,12a-tetraazabenzo[4,5]cyclohepta[1,2,3-de]naphthalen-5-yl]naphthalen-2-ol

[0377] [Chemical Structure]

[0378] Preparation Example 9 (450 mg, 1.51 mmol) was suspended in CH3CN (10 mL). DIPEA (276 uL, 1.59 mmol) was added and the suspension was cooled to 0 °C under N2. In a separate vial, CAS 1262409-55-1-HCl salt (232 mg, 1.39 mmol) was suspended in DCM (1 mL), and DIPEA (276 uL, 1.59 mmol) was added to dissolve the amine-HCl salt. THF (6 mL) was added to the resulting solution to obtain an emulsion mixture. This solution was added to the flask containing the cold solution of Preparation Example 9. After about 45 minutes at 0 °C, LCMS analysis indicated that the initial reaction was complete. LiOtBu (1 M in THF 4.5 mL, 4.5 mmol) was added dropwise and the ice bath was removed. The ice bath was replaced with an oil bath and the reaction was heated at 50 °C for 30 minutes. LCMS analysis indicated that the cyclization step was complete. The solution was cooled to room temperature and evaporated. Saturated aqueous NaHCO3 (10 mL) was added and the mixture was extracted with DCM (3 × 30 mL). The combined organic extracts were dried over Na2SO4 and evaporated. The process described above was repeated again on the same scale with the same observations and results. The crude materials from both reactions were combined and purified using flash chromatography eluting with a gradient of 0 - 100% EtOAc in heptane, loading the crude material onto a silica cartridge using DCM. The fractions containing the desired product were pooled and concentrated to give 483 mg (70%) of 14a as a light brown solid. 1 H NMR (400 MHz, chloroform-d) δ = 5.22 (ddd, J = 2.9, 6.8, 13.8 Hz, 1H), 4.65 (dd, J = 4.6, 13.4 Hz, 1H), 4.42 (d, J = 13.4 Hz, 1H), 4.22 - 4.17 (m, 1H), 4.09 - 3.98 (m, 2H), 3.72 (dd, J = 9.8, 12.6 Hz, 1H), 3.43 - 3.24 (m, 2H), 2.62 (s, 3H), 2.26 - 2.12 (m, 1H), 2.05 - 1.92 (m, 1H). 19 F NMR (376 MHz, chloroform-d) δ = -140.51 (s, 1F). Analysis of this substance by chiral SFC showed an optical purity of 50% ee, indicating that the starting amino alcohol (CAS 1262409-55-1-HCl salt) was not optically pure. In a subsequent sequence, the minor enantiomer was removed using preparative chiral SFC (see below). To a vial equipped with a stir bar was added 14a (400 mg, 1.12 mmol), CAS 2621932-37-2 (689 mg, 1.35 mmol), K2CO3 (465 mg, 3.4 mmol) added as a solution in 1,4-dioxane (5.6 mL) and water (0.6 mL). The mixture was purged with N2 for 3 minutes and RuPhos Pd G3 (94 mg, 0.11 mmol) was added. The vial was sealed and heated at 90 °C for 4.5 hours. LCMS indicated consumption of the starting material and water (5 mL) was added. The dark mixture was extracted with EtOAc (3 × 30 mL). The combined organic extracts were dried over Na2SO4, filtered, evaporated, and subjected to chiral SFC purification to remove the minor enantiomer. After chiral SFC, 480 mg (62%) of 14b was obtained at 90% ee. 1 1H NMR (400 MHz, methanol-d4 ) δ = 7.95 (dd, J = 5.8, 9.1 Hz, 1H), 7.64 (d, J = 2.5 Hz, 1H), 7.40 (t, J = 8.9 Hz, 1H), 7.34 (dd, J = 2.3, 19.4 Hz, 1H), 5.39 - 5.30 (m, 2H), 5.26 - 5.14 (m, 1H), 4.71 (ddd, J = 4.7, 13.6, 18.6 Hz, 1H), 4.57 - 4.48 (m, 1H), 4.31 (tt, J = 4.9, 9.4 Hz, 1H), 4.24 - 4.14 (m, 1H), 4.06 - 3.79 (m, 2H), 3.71 - 3.52 (m, 2H), 3.51 - 3.48 (m, 3H), 2.61 (s, 3H), 2.28 - 2.12 (m, 1H), 2.07 - 1.88 (m, 1H), 0.98 - 0.90 (m, 18H), 0.77 - 0.61 (m, 3H), 19 19F NMR (376 MHz, methanol-d4) δ = -108.33 (d, J = 69.4 Hz, 1F), -141.66 - -144.80 (m, 1F), MS: 707 [M+H] + 14b (402 mg, 0.57 mmol) was dissolved in DMF (9.5 mL). Anhydrous CsF (518 mg, 3.42 mmol) was added and the reaction was heated to 35 °C for 45 minutes. Most of the DMF was removed in vacuo (5 mm Hg, 30 °C) and water (5 mL) was added to the resulting residue. A solid precipitated and it was extracted with DCM (3 × 20 mL). The combined organic extracts were dried over Na2SO4, filtered and evaporated to give a crude product which was subjected to chiral SFC to further enrich the desired enantiomer. After chiral SFC, 257 mg (82%) of 14c was obtained as a light brown solid. 1 1H NMR (400 MHz, chloroform-d) δ = 7.87 - 7.74 (m, 1H), 7.55 - 7.50 (m, 1H), 7.46 - 7.35 (m, 1H), 5.37 - 5.16 (m, 3H), 4.71 - 4.59 (m, 1H), 4.54 - 4.41 (m, 1H), 4.21 (dt, J = 3.8, 12.2 Hz, 1H), 4.14 - 3.97 (m, 2H), 3.83 (td, J = 9.1, 12.6 Hz, 1H), 3.52 (d, J = 2.8 Hz, 3H), 3.47 - 3.23 (m, 2H), 2.92 (d, J = 19.6 Hz, 1H), 2.64 (s, 3H), 2.36 - 2.17 (m, 1H), 2.13 - 1.93 (m, 1H), 1.50 (s, 1H), 1919F NMR (376 MHz, chloroform-d) δ = -106.89 - -109.75 (m, 1F), -143.05 (d, J = 363.4 Hz, 1F). A mixture of 14c (252 mg, 0.46 mmol) in MEK (7 mL) was added to solid oxone (646 mg, 1.03 mmol) and saturated aqueous NaHCO3 solution (7 mL). The mixture was stirred at 1500 rpm for 20 minutes at room temperature. The reaction was diluted with EtOAc (20 mL) and the aqueous layer was further extracted twice with EtOAc (2 × 20 mL). The combined organic extracts were washed with 10% aqueous Na2S2O3 solution (10 mL) and brine (10 mL). After drying over Na2SO4, the mixture was filtered and evaporated to give 301 mg of the sulfone as a pale yellow rubbery solid. MS: 583 [M+H] + This material was taken to the next step without further purification. The sulfone (267 mg, 0.458 mmol) and CAS 2097518-76-6 (100 mg, 0.64 mmol) were dissolved in CH3CN (0.9 mL). Lithium trimethylsilanolate (130 mg, 1.4 mmol) was added, the vial was sealed, stirred and heated to 50 °C for 30 minutes to give 14d, which was taken to the MOM deprotection step in the same pot. After cooling to room temperature, MeOH (2 mL) was added to the 14d solution, followed by HCl (4 M in 1,4-dioxane 3.5 mL, 14 mmol) and the reaction was stirred at room temperature for 30 minutes. The solvent was removed under vacuum and saturated aqueous NaHCO3 solution (10 mL) and water (10 mL) were added. A solid precipitated from this mixture, which was insoluble in EtOAc but soluble in DCM. The aqueous mixture was extracted with DCM 1 × 150 mL, then 2 × 50 mL. The combined organic extracts were dried over Na2SO4, filtered, evaporated and subjected to purification. Purification was achieved using SFC eluting with CO2 / MeOH 10 - 50% at 120 bar, 100 mL / min on a Princeton HA-Morpholine 5um 21.2 × 150 mm column over 5.0 minutes to give 193 mg (68%) of Example 33 as a white solid. 11H NMR (400 MHz, methanol-d4) δ = 7.83 (dd, J = 5.7, 9.1 Hz, 1H), 7.32 (d, J = 2.5 Hz, 1H), 7.32 - 7.27 (m, 1H), 7.25 - 7.15 (m, 1H), 5.41 - 5.17 (m, 2H), 4.72 (td, J = 4.6, 13.5 Hz, 1H), 4.61 - 4.53 (m, 1H), 4.41 - 4.22 (m, 3H), 4.22 - 4.11 (m, 1H), 4.04 - 3.92 (m, 1H), 3.82 (ddd, J = 5.3, 9.8, 12.3 Hz, 1H), 3.71 - 3.51 (m, 1H), 3.50 and 3.36 (alkyne H has two chemical shifts, both doublets, J = 0.8 Hz, 1H), 3.49 - 3.38 (m, 1H), 3.28 - 2.95 (m, 4H), 2.42 - 2.09 (m, 4H), 2.06 - 1.81 (m, 4H); 19 19F NMR (377 MHz, methanol-d4) δ = - 111.68 (qd, J = 4.6, 61.0 Hz, 1F), - 143.97 - - 147.51 (m, 1F), - 173.14 - - 174.37 (m, 1F), MS: 618 [M+H] + .

[0379] (Example 34) ({5-Ethynyl-6-fluoro-4-[(8aS)-4-fluoro-2-{[(2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl]methoxy}-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazaphenanthro[1,8-ab]heptalen-5-yl]naphthalen-2-yl}oxy)methyl dihydrogen phosphate.

[0380]

Chem.

[0381] Example 34 was developed as a prodrug of Example 33 to improve the unbound exposure of Example 33 in plasma.

[0382] The solution formulation of Example 33 was an aqueous solution containing 2.5% (w / v) of Pluronic F-68 (Poloxamer 188). The amorphous suspension formulations of Example 33 and Example 34 were prepared using 0.5% (w / v) methylcellulose in water.

[0383] As demonstrated in Figure 1 and Table 1-A, after a single oral dose of 100 mg (active) / kg in female NSG mice, the mean systemic exposure of Example 33 (evaluated by AUC and / or Cav) was approximately 10 - 17 times higher after administration of Example 34 compared to the oral suspension dose of Example 33 (free base).

[0384]

Table 3

[0385] Predicted deuterated analog (PDA) of Example 33 The compounds presented in Table 2 are the predicted deuterated analogs (PDA) of Example 33. Formula (V) is the general formula of deuterated Example 33, where Y 1a , Y 1b , Y 2a , Y 2b , Y 3a , Y 3b , Y 4 and Y 5 are each independently H or D. The deuterated analogs of Example 33 in Table 2 were predicted based on the metabolic profile of Example 33 using MetaSite (moldiscovery.com / software / metasite / ). Y 1a , Y 1b , Y 2a , Y 2b , Y 3a , Y 3b , Y 4 and Y 5 are most likely to be the positions where metabolism occurs based on the MetaSite prediction.

[0386]

Chemical formula

[0387]

Table 4

[0388] General methods / outlines for obtaining metabolite profiles of compounds and identifying metabolites are described in Dalvie et al., "Assessment of Three Human in Vitro Systems in the Generation of Major Human Excretory and Circulating Metabolites", Chemical Research in Toxicology, 2009, 22, 2, 357-368, tx8004357 (acs.org); King, R., "Biotransformations in Drug Metabolism", Ch.3, Drug Metabolism Handbook Introduction, https: / / doi.org / 10.1002 / 9781119851042.ch3; Wu, Y. et al., "Metabolite Identification in the Preclinical and Clinical Phase of Drug Development", Current Drug Metabolish, 2021, 22, 11, 838-857, 10.2174 / 1389200222666211006104502; Godzien, J. et al., "Chapter Fifteen - Metabolite Annotation and Identification".

[0389] To assist in predicting the metabolic pathways and metabolites of compounds, a number of publicly available commercial software tools are available. Examples of such tools include BioTransformer 3.0 (biotransformer.ca / new) (which predicts the in vivo biotransformation of small molecules using a database of known metabolic reactions); MetaSite (moldiscovery.com / software / metasite / ) (which predicts cytochrome P450-related metabolic conversions and flavin-containing monooxygenase-mediated reactions in phase I metabolism); and Lhasa Meteor Nexus (lhasalimited.org / products / meteor-nexus.htm) (which uses a wide range of machine learning models to propose predictions of metabolic pathways and metabolite structures and covers phase I and phase II in vivo biotransformations of small molecules).

[0390] The predicted deuterated analogs V-1 to V-18 of Example 33 in Table 2 may result in certain therapeutic advantages arising from greater metabolic stability, such as an increase in in vivo half-life, a reduction in dosing requirements, a decrease in CYP450 inhibition (competitive or time-dependent), or an improvement in the therapeutic index or tolerance.

[0391] One of ordinary skill in the art would 1a , Y 1b , Y 2a , Y 2b , Y 3a , Y 3b , Y 4 and Y 5 Additional deuterated analogs of Example 33 having various combinations of Y

[0392] KRAS surface plasma resonance (SPR) binding assay The kinetic binding constant (K D ) of the examples of the present invention was measured using an SPR assay.

[0393] The binding affinity and kinetics of the examples of the present invention were measured by surface plasmon resonance (SPR) using a Biacore 8K or 8K+ (Cytiva, Marlborough, MA) instrument. In these experiments, recombinant C-terminal site-specific biotinylated wild-type (WT) KRAS (aa1-185), G12D KRAS (aa1-185), G12C KRAS (aa2-184), G12V KRAS (aa2-184), WT HRAS (aa2-184), and WT NRAS (aa2-185) proteins purified in the presence of 1 μM GDP were used. Binding measurements were performed in parallel in either set of WT / G12D / G12C / G12V KRAS or WT K / H / N RAS proteins.

[0394] The Biacore instrument was detached and docked with a Series S Sensor Chip SA. The protein was diluted to 50 μg / mL with assay buffer (50 mM HEPES, 150 mM NaCl, 10 μM GDP, 5 mM MgCl2, 0.5 mM TCEP, 5% glycerol, 0.02% Tween-20, 2% DMSO, pH 7.2) and immobilized at a flow rate of 3 μL / min at 10 °C for a contact time of 15 minutes to capture approximately 3000 - 4000 RU of the protein on the surface. The functionalized surface was then equilibrated with assay buffer for approximately 1 hour. The non-functionalized SA surface without immobilized protein served as a reference for binding kinetic analysis. Compound binding kinetics were measured in either multi-cycle or single-cycle kinetic formats.

[0395] Multi-cycle kinetic analysis (MCK) A two-fold 10-point serial dilution of the test compound was prepared in a 96-well microplate (Greiner; Cat # 650101) at either a maximum concentration of 10 μM or 100 μM. At 10 °C, the serial dilutions of the compound were injected into both the reference and RAS immobilized channels at a flow rate of 100 μL / min and an association time of 90 seconds to measure the binding kinetics. Compound dissociation was monitored for at least 400 seconds during each cycle. No additional regeneration was used. DMSO calibration curves were obtained by injecting 0 - 4% DMSO in the assay buffer before and after compound analysis. As a positive control to assess the activity of the protein captured on the surface, a suitable compound with known affinity and kinetics was tested once in each experiment.

[0396] Single Cycle Kinetic Analysis (SCK) A three-fold 6-point serial dilution of the compound was prepared in a deep well 96-well microplate (Greiner Bio; Cat # 780201) at a maximum concentration of 1 μM (concentration range: 0.004 - 1 μM). At 10 °C, the serial dilutions of the compound were injected into the reference and further into the RAS immobilized channel in ascending order at a flow rate of 100 μL / min and an association time of 120 seconds to measure the binding kinetics. Compound dissociation was monitored for at least 3600 seconds. Two buffer blanks were also run in single cycle kinetics format, followed by the compound for double referencing. No additional regeneration was used. DMSO calibration curves were obtained by injecting 0 - 4% DMSO in the assay buffer before and after compound analysis. As a positive control to assess the activity of the protein captured on the surface, a suitable compound with known affinity and kinetics was tested once in each experiment.

[0397] Both MCK and SCK data were processed and analyzed using Biacore Insight Evaluation Software (Cytiva, Marlborough, MA). Double-reference and solvent-corrected data were fitted to a 1:1 Langmuir model to measure the kinetic binding constant (K D ), the association rate (k on ), and the dissociation rate (k off ). The dissociation half-life (t 1 / 2 ) was calculated from the measured k off using the standard equation (t 1 / 2 = 0.693 / k off ). The validity of the fit was judged by the c 2 value and the randomness of the residue distribution.

[0398] The SPR binding assay results in Examples 1 to 31 are presented in Table 3. When an example was tested more than once, the geometric mean of the binding constant K D was presented (n is the number of assay replicates). The blanks in Table 3 indicate that no data were obtained for that example in a particular assay.

[0399] The binding constant K D indicates that the exemplified compounds can have strong binding ability to all of the KRAS G12C, KRAS G12D, and KRAS G12V receptors and selectivity over the HRAS and NRAS receptors.

[0400]

Table 5

[0401] KRAS Cell Titer Glo (CTG) assay The CellTiter-Glo® (CTG) Luminescent Cell Viability Assay is a homogeneous method for determining the number of viable cells in a culture based on the quantification of existing ATP, which is an indicator of metabolically active cells. CTG is designed for use in multi-well formats and is ideal for automated high-throughput screening (HTS), cell proliferation, and cytotoxicity assays. The homogeneous assay procedure involves directly adding a single reagent (CellTiter-Glo® reagent) to cells cultured in serum-supplemented media. No cell washing, medium removal, or multiple pipetting steps are required. This system can detect as few as 15 cells per well in a 384-well format within 10 minutes of reagent addition and mixing.

[0402] The cells are grown in a humidified 5% CO2 incubator at 37°C using the culture conditions outlined below. All cell culture media reagents were purchased from Gibco. The cell lines were purchased from ATCC: H358 (non-small cell lung cancer cell line), SW620 (colorectal cancer cell line), PANC08.13 (RPMI1640 + 10% FBS + insulin 10 units / ml, pancreatic cancer cell line). Test and control compounds are dispensed onto 384 assay plates (Corning, Cat#3764) as nanoliter drops at the desired final concentration in 0.1% DMSO using an Echo Acoustic Dispenser, and then the cells are seeded. The cells are seeded at the following cell densities (number of cells per well) in a volume of 40 μL per well: H358 (300), SW620 (750), PANC 08.13 (600). The cells are incubated for 7 days in the presence of the compounds. The viability is determined on day 7 using the CellTiter-Glo® (CTG) Luminescent Cell Viability Assay (Promega). CTG is added to a final volume of 20 μl per well and incubated at room temperature for 15 minutes, and then the luminescence is captured using an EnVision Reader with the LUM384 US protocol. The data are analyzed using Activity Base to determine the compound response and presented as either percent activity (PCTEFF) or percent control (PCTOCTL) as described: zero percent activity control (ZPE) (negative control) - DMSO 100%. Hundred percent activity (HPE) (positive control) - 1 μM trametinib (GSK1120212, MEK inhibitor) (10 mM 4 nl per well and DMSO 36 nl). The following formula / nomenclature is used (percent activity; PCTEFF) and (percent of control; PCTOCTL): PCTEFF: 100×(raw data value - HPE / ZPE - HPE), PCTOCTL: 100×raw data value / User_Defined_Array), where User_Defined_Array is either the aggregated HPE or ZPE.

[0403] The CTG assay results for some of the illustrated examples are presented in Table 4. When the examples were tested more than once, the geometric mean of IC 50 (nM) was presented (n is the number of test replicates).

[0404] The CTG assay indicates that the exemplary compounds of the present invention selected have anti-cancer activity demonstrated for pancreatic cancer, non-small cell lung cancer, and colorectal cancer.

[0405] [Table 6]

[0406] It will be apparent to those skilled in the art that the present invention can be variously modified and varied without departing from the scope or spirit of the present invention. By considering the specification and implementation of the present invention disclosed herein, other embodiments of the present invention will be apparent to those skilled in the art. The specification and examples are to be considered as illustrative only, and it is intended that the true scope and spirit of the present invention be indicated by the following claims.

[0407] All references cited herein, including patents, patent applications, documents, textbooks, and the like, as well as references cited therein, are hereby incorporated by reference in their entirety herein to the extent not already incorporated. To the extent that one or more of the incorporated documents and the like, including defined terms, usage of terms, described techniques, or the like, are different from or conflict with the present application, the present application shall control.

Claims

1. A compound of formula (V): 【Chemical Formula 1】 Or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable additive, a pharmaceutical composition [Wherein, R １ is 【Chemical 2】 Selected from the group consisting of R ２ is C １ alkyl, C ３ alkyl, -(C １ alkylene)-OH, or -(C ３ alkylene)-OH, and R ３ is 【Chemical Formula 3】 Selected from the group consisting of R ４ is Cl or F, R ５ is -(C １ alkylene)-OH, or C １ alkyl, and R ２ and R ５ may together form a 7-membered heterocycloalkyl containing one heteroatom O R ６ represents one or two substituents selected from the group consisting of H, -OH, halogen, -(C １ ~C ６ alkylene)-OH, -CN, -(C １ ~C ６ alkylene)-CN, C １ ~C ６ alkyl, C ３ ~C ６ cycloalkyl, C １ ~C ６ fluoroalkyl, C ３ ~C ６ fluorocycloalkyl, and C １ ~C ６ alkoxy, and L is a linker containing 1, 2 or 3 members independently selected from the group consisting of -O-, -S-, -NR ７ -, and -CR ８ R ９ -, and is a linker containing 1, 2 or 3 members independently selected from the group consisting of R ７ 、 R ８ 、 and R ９ are each independently H or C １ ~C ３ alkyl, X is O, N, or S, l is 1 or 2].

2. R １ is [Chemical Formula 4] Selected from the group consisting of, the pharmaceutical composition according to claim 1.

3. R １ is 【Chemical 5】 Is, the pharmaceutical composition according to claim 2.

4. R ２ is C ３ alkyl, and R ５ is -(C １ alkylene)-OH, the pharmaceutical composition according to claim 1.

5. R ２ is -(C ３ alkylene)-OH, and R ５ is C １ alkyl, the pharmaceutical composition according to claim 1.

6. R ２ and R ５ The pharmaceutical composition according to claim 4, wherein R and R together form a 7-membered heterocycloalkyl containing one heteroatom O.

7. R ３ is 【Chemical Formula 6】 Is, the pharmaceutical composition according to claim 1.

8. R ４ The pharmaceutical composition according to claim 1, wherein R is F.

9. Where X is O and l is 1, the pharmaceutical composition according to claim 1.

10. The compound of formula (V) is a compound of formula (VI): 【Chemical Formula 7】 Having the compound, the pharmaceutical composition according to claim 1.

11. R ２ is C ３ alkyl, and R ５ is -(C １ alkylene)-OH, the pharmaceutical composition according to claim 10.

12. R ２ is -(C ３ alkylene)-OH, and R ５ is C １ alkyl, the pharmaceutical composition according to claim 10.

13. R ２ and R ５ The pharmaceutical composition according to claim 10, wherein R and R together form a 7-membered heterocycloalkyl containing one heteroatom O.

14. A compound of formula (VII) [Chemical Formula 8] Or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable additive, a pharmaceutical composition [wherein, R １１ , R １２ , R １３ , and R １４ are each independently H or C １ -C ３ alkyl].

15. R １１ 、 R １２ 、 R １３ 、 and R １４ is, independently of one another, H or methyl, the pharmaceutical composition according to claim 14.

16. The compound of formula (VII) is the following formula: 【Chemical Formula 9】 Having the compound, the pharmaceutical composition according to claim 14.

17. 【Fig. 10】 A pharmaceutical composition comprising a compound selected from the group consisting of or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable additive.

18. For use in treating cancer, the pharmaceutical composition according to any one of claims 1 to 17.

19. For use in combination with an additional anti-cancer therapeutic agent, the pharmaceutical composition according to claim 18.

20. Wherein the cancer is non-small cell lung cancer (NSCLC), pancreatic cancer, or colorectal cancer, the pharmaceutical composition according to claim 18.

21. A pharmaceutical composition according to claim 19 for use in treating cancer, wherein the cancer is non-small cell lung cancer (NSCLC), pancreatic cancer, or colorectal cancer, a pharmaceutical composition.

22. For use in treating disorders mediated by inhibition of the KRAS G12C, KRAS G12D, and KRAS G12V receptors in a subject, the pharmaceutical composition according to any one of claims 1 to 17.

23. Further comprising at least one additional therapeutic agent or a pharmaceutically acceptable salt thereof, the pharmaceutical composition according to claim 18.

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