Pyrido[4,3-d]pyrimidine compounds

Abstract

This invention relates to compounds of formulas (I) to (III) and their pharmaceutically acceptable salts, their use in pharmaceuticals; compositions containing them; processes for preparing them; and intermediates used in such processes. The compounds of this invention may be useful in the treatment, prevention, suppression, and remission of diseases, disorders, and conditions such as cancer.

Description

[Technical Field]

[0001] This invention relates to a novel pyrido[4,3-d]pyrimidine compound as a Kirsten rat sarcoma virus oncogene homolog (KRAS) inhibitor. The invention also relates to the preparation of the compound, intermediates used in its preparation, compositions containing the compound, and the use of the compound for treating KRAS-related diseases such as cancer. [Background technology]

[0002] KRAS, HRAS (Harvey rat sarcoma virus), and NRAS (neuroblastoma RAS virus oncogene homolog) belong to a group of GTPases that are crucial for cell survival and proliferation through complex signaling cascades. Mutations in the RAS gene 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 in cancer cells (up to 85%), leading to the development of cancers including non-small cell lung cancer (NSCLC), colorectal cancer, and pancreatic cancer, which represent a significant unmet medical need for affected patients, both in populations and individuals. KRAS mutations are widely found in pancreatic tubular adenocarcinoma (PDAC). Mutations in KRAS have been observed in 30% of NSCLC cases, the dominant form of lung cancer (80%). KRAS mutations found in NSCLC include G12C (39%), G12V (18-21%), and G12D (17-18%). KRAS mutations occur in 35-45% of colon cancers and lead to drug resistance.

[0003] KRAS inhibitors have been sought after for decades, and recent advances include the approval of sotrasib and subsequent KRAS G12C-targeting compounds in trials (Palmer et al., 2021 NPJ Precision Oncology, 5, 98). Sotrasib specifically targets KRAS mutations through covalent modification of the mutant cysteine ​​at position 12. Therefore, sotrasib and other currently known KRAS inhibitors that rely on the same mechanism of action may have a narrow therapeutic range and limited use when considering other major KRAS mutations such as G12V and G12D.

[0004] The majority of active KRAS mutant inhibitors capable of targeting mutations other than G12C obtain their proper binding efficacy through phenolic components. Phenols are prone to conjugation metabolism in the intestine and liver, which can lead to reduced bioavailability. Therefore, phenol-less chemiforms may have advantages in terms of bioavailability and drug delivery (Fell et al., 2020 J.Med.Chem 63, 6679-6693). [Overview of the project] [Problems that the invention aims to solve]

[0005] Therefore, there is still a need for new KRAS inhibitors that can be used to treat a wider range of cancers. [Means for solving the problem]

[0006] The present invention provides, in part, compounds of formulas (I), (II), and (III), as well as pharmaceutically acceptable salts thereof. The compounds of the present invention can inhibit all activity of the KRAS G12C, KRAS G12D, and KRAS G12V receptors and may be useful in the treatment, prevention, suppression, and remission of diseases such as cancer, disorders, and pathologies mediated by any of the KRAS G12C, KRAS G12D, and KRAS G12V receptors, or combinations thereof. In particular, the present invention provides compounds containing indazole components that have unexpectedly high cytotoxicity against one or more KRAS G12C, KRAS G12D, and KRAS G12V receptors. Pharmaceutical compositions comprising the compounds or salts of the present invention alone or in combination with additional anticancer agents are also provided. 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 for using the above. This summary is provided to introduce, in a simplified form, selected from the concepts further described below in the detailed description. This summary is not intended to identify any material or essential features of the subject matter described in the claims, nor is it intended to be used in isolation as an aid in determining the scope of the subject matter described in the claims.

[0007] According to one embodiment of the present invention, a compound of formula (I):

[0008] [ka] or to provide a pharmaceutically acceptable salt thereof. [In the formula, R 1 C3~C 10 A 4-12 membered heterocycloalkyl containing 1, 2, or 3 heteroatoms selected from the group consisting of cycloalkyl or N, O, and S, and C3-C 10Cycloalkyl or 4- to 12-member heterocycloalkyl may each be substituted with 1, 2 or 3 substituents independently selected from the group consisting of -OH, -CN, halogen, C1-C3 alkylidenyl, C1-C3 haloalkylidenyl, C1-C3 alkyl (where, if present, two of the C1-C3 alkyls may together with the carbon to which they are attached form a spirocyclic ring), C1-C3 alkoxy, -OC(O)NH2, -OC(O)NHCH3, -OC(O)N(CH3)2, and C1-C3 alkyl, C1-C3 alkoxy, C1-C3 haloalkylidenyl or C1-C3 alkylidenyl may each be further substituted with 1, 2 or 3 11 R substituents, R 2 is H or is -(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, or C1-C6 alkoxy, each of which may be independently substituted with 1, 2 or 3 substituents selected from the group consisting of -OH, -CN, -NH2, -NH(C1-C3 alkyl), -N(C1-C3 alkyl)2, -SH, -(C1-C4 alkylene)-CN, -(C1-C4 alkylene)-OH, halogen, C1-C3 alkyl, C3-C6 cycloalkyl, C1-C3 fluoroalkyl, and C1-C3 alkoxy, R 3 is

[0009]

Chemical formula

[0010] Embodiments of the present invention are described below, but for convenience, Embodiment 1 (E1) is identical to the embodiment of formula (I) presented above.

[0011] It should be understood that both the general description above and the following detailed description are illustrative and descriptive only, and do not limit the invention as described in the claims. [Modes for carrying out the invention]

[0012] The present invention can be more readily understood by referring to the following detailed description of embodiments and examples of the present invention included herein. It should be understood that the present invention is not limited to specific synthesis methods, and that synthesis methods may, of course, vary. It should also be understood that the technical terms used herein are for the sole purpose of describing specific embodiments and are not intended to be limiting.

[0013] E1 A compound of formula (I) as defined above, or a pharmaceutically acceptable salt thereof.

[0014] E2 A compound of Embodiment E1, or a pharmaceutically acceptable salt thereof, wherein the linker L is -(O-CH2)-.

[0015] E3 R 1 The 5-10 member heterocycloalkyl is a 5-10 member heterocycloalkyl containing one or two heteroatoms selected from the group consisting of N and O, and the 5-10 member heterocycloalkyl may be substituted with one, two or three substituents independently selected from the group consisting of -OH, -CN, halogen, C1-C3 alkylidenyl, C1-C3 haloalkylidenyl, C1-C3 alkyl (where, if present, two of the C1-C3 alkyls may form a spirocyclic ring together with the carbons to which they are bonded), C1-C3 alkoxy, -OC(O)NH2, -OC(O)NHCH3, or -OC(O)N(CH3)2, and the C1-C3 alkyl, C1-C3 alkoxy, C1-C3 haloalkylidenyl or C1-C3 alkylidenyl contains one, two or three R 11 Compounds of Embodiment E1 or Embodiment E2, or pharmaceutically acceptable salts thereof, which may be further substituted with substituents, respectively.

[0016] E3a LR 1 However, it may be substituted with one, two, or three substituents independently selected from the group consisting of -OH, -CN, halogen, C1-C3 alkylidenyl, C1-C3 haloalkylidenyl, C1-C3 alkyl (where, if present, two of the C1-C3 alkyls may form a spirocyclic ring together with the carbons to which they are bonded), C1-C3 alkoxy, -OC(O)NH2, -OC(O)NHCH3, and -OC(O)N(CH3)2.

[0017] [ka] The C1-C3 alkyl, C1-C3 alkoxy, C1-C3 haloalkylidenyl, or C1-C3 alkylidenyl is present in 1, 2, or 3 R groups. 11Compounds of Embodiment E1 or Embodiment E2, or pharmaceutically acceptable salts thereof, which may be further substituted with substituents, respectively.

[0018] E3b LR 1 However, it may be substituted with one, two, or three halogens.

[0019] [ka] A compound of Embodiment E1 or Embodiment E2, or a pharmaceutically acceptable salt thereof.

[0020] E3c LR 1 However, it may be substituted with one or two substituents independently selected from the group consisting of -OH, -CN, halogens, and C1-C3 alkyl groups.

[0021] [ka] And alkylidenyl has 1 or 2 R 11 Compounds of Embodiment E1 or Embodiment E2, or pharmaceutically acceptable salts thereof, which may be further substituted with substituents.

[0022] E3d LR 1 However, it may be substituted with one or two substituents independently selected from the group consisting of -OH, -CN, halogens, and C1-C3 alkyl groups.

[0023] [ka] And R 11 However, H, -CN, -OH, methyl, -OCH3, C1~C3 alkoxy, -cyclopropyl, -oxetane, -C(O)NR 7 R 8 , -SO2R 9 and halogens are independently selected from the group, or R 11The compounds of Embodiment E1 or Embodiment E2, or pharmaceutically acceptable salts thereof, wherein they form a C3-C6 cycloalkyl ring or a 3-6 membered heterocycloalkyl ring together with the carbon atoms to which they are bonded.

[0024] E3e LR 1 However, it may be substituted with one or two substituents independently selected from the group consisting of -OH, -CN, halogens, and C1-C3 alkyl groups.

[0025] [ka] The compound of embodiment E3d, or a pharmaceutically acceptable salt thereof.

[0026] E4 LR 1 but,

[0027] [ka] A compound of Embodiment E3, or a pharmaceutically acceptable salt thereof, selected from the group consisting of the following.

[0028] E4b LR 1 but,

[0029] [ka] A compound of Embodiment E3, or a pharmaceutically acceptable salt thereof, selected from the group consisting of the following.

[0030] E5 LR 1 but,

[0031] [ka] A compound of Embodiment E3, or a pharmaceutically acceptable salt thereof, selected from the group consisting of the following.

[0032] E6 LR 1 but,

[0033] [ka] A compound of Embodiment E3, or a pharmaceutically acceptable salt thereof, selected from the group consisting of the following.

[0034] E7 R 2 A compound selected from the group consisting of -(C1-C5 alkylene)-OH and C1-C5 alkyl, or a pharmaceutically acceptable salt thereof, which is H, or which may be substituted with one, two, or three substituents independently selected from the group consisting of -OH, -CN, and halogens.

[0035] E8 R 3 However, 1, 2, or 3 R 10 It may be replaced with

[0036] [ka] A compound from any one of embodiments E1 to E7, or a pharmaceutically acceptable salt thereof.

[0037] E9 R 3 but,

[0038] [ka] A compound from any one of embodiments E1 to E8, or a pharmaceutically acceptable salt thereof.

[0039] E9a R 3 but,

[0040] [ka] A compound from any one of embodiments E1 to E8, or a pharmaceutically acceptable salt thereof.

[0041] E9b R 3 but,

[0042] [ka] And R 10 A compound from any one of Embodiments E1 to E8 or E9a, or a pharmaceutically acceptable salt thereof, which is independently selected from the group consisting of 4-6 membered heterocycloalkyls containing one or two heteroatoms selected from the group consisting of -NH2, halogen, -CN, C1-C3 alkyl, C1-C3 fluoroalkyl, C1-C3 alkoxy, C2-C3 alkynyl, C3-C5 cycloalkyl, and N, O, and S, wherein the C1-C3 alkyl, C3-C5 cycloalkyl, or 4-6 membered heterocycloalkyl may be further substituted with one or two substituents independently selected from the group consisting of -OH, C1-C3 alkoxy, -C1-C3 alkyl, and halogen.

[0043] E10 R 3 but,

[0044] [ka] A compound from any one of embodiments E1 to E9, or a pharmaceutically acceptable salt thereof.

[0045] E10a R 3 but,

[0046] [ka] The compound is one of the embodiments of E9a to E9b, or a pharmaceutically acceptable salt thereof.

[0047] E11 R 3 However, one R 10 Or 1, 2, or 3 R 12 It may be further replaced with

[0048] [ka] A compound from any one of embodiments E1 to E9a, or a pharmaceutically acceptable salt thereof.

[0049] E11a R 3 but,

[0050] [ka] The compound of Embodiment E11, or a pharmaceutically acceptable salt thereof.

[0051] E11b R 3 but,

[0052] [ka] The compound of Embodiment E11, or a pharmaceutically acceptable salt thereof.

[0053] E11c R 3 but,

[0054] [ka] The compound of Embodiment E11, or a pharmaceutically acceptable salt thereof.

[0055] E11d R 3 but,

[0056] [ka] The compound of Embodiment E11, or a pharmaceutically acceptable salt thereof.

[0057] E11e R 3 but,

[0058] [ka] A compound of embodiment E11 or E11b, or a pharmaceutically acceptable salt thereof.

[0059] E11f R 3 but,

[0060] [ka] A compound of embodiment E11 or E11b, or a pharmaceutically acceptable salt thereof.

[0061] E11g R 3 but,

[0062] [ka] A compound of embodiment E11 or E11b, or a pharmaceutically acceptable salt thereof.

[0063] E11h R 3 but,

[0064] [ka] A compound of embodiment E11 or E11b, or a pharmaceutically acceptable salt thereof.

[0065] E12 R 4 The compound is one of any one of embodiments E1 to E11h, or a pharmaceutically acceptable salt thereof, wherein the compound is Cl or F.

[0066] E13 R 5 and R 6 However, each time it appears, it is independently selected from the group consisting of H, -OH, -CN, and halogens, or R 5 and R 6 Each occurrence of -(C1-C5 alkylene)-OH and C1-C5 alkyl may be independently selected from the group consisting of -(C1-C5 alkylene)-OH and C1-C5 alkyl, and each of -(C1-C5 alkylene)-OH and C1-C5 alkyl may be substituted with one, two, or three substituents independently selected from the group consisting of -OH, -CN, and halogen, one of the compounds from Embodiments E1 to E12, or a pharmaceutically acceptable salt thereof.

[0067] E14 Formula (II):

[0068] [ka] [In the formula, Y is selected from the group consisting of CH2, O, N(C1~C6 alkyl), S, (S=O), and (SO2), R 13 Each instance of -OH, -CN, halogen, C1-C3 alkyl, -(C1-C6 alkylene)-CN, and -(C1-C6 alkylene)-OH is independently selected from the group, m and n are independently 0, 1, 2, or 3, y is 1, 2, or 3, and m+n is 1, 2, 3, 4, or 5. A compound from any one of embodiments E1 to E13, or a pharmaceutically acceptable salt thereof, having the above characteristics.

[0069] E15 A compound of Embodiment E14, or a pharmaceutically acceptable salt thereof, wherein Y is -CH2- or O.

[0070] E16 A compound of Embodiment E15, or a pharmaceutically acceptable salt thereof, wherein Y is O.

[0071] E17 R3 but,

[0072] [ka] A compound selected from the group consisting of E1 to E8 or E14 to E16, or a pharmaceutically acceptable salt thereof.

[0073] E17a R 3 but,

[0074] [ka] A compound selected from the group consisting of E1 to E8 or E14 to E16, or a pharmaceutically acceptable salt thereof.

[0075] E17b R 3 but,

[0076] [ka] A compound selected from the group consisting of E1 to E8 or E14 to E16, or a pharmaceutically acceptable salt thereof.

[0077] E18 Formula (III):

[0078] [ka] A compound from any one of embodiments E1 to E17b, or a pharmaceutically acceptable salt thereof, having the above characteristics.

[0079] E18a

[0080] [ka] Not any one of the compounds of Embodiments E1 to E17, or a pharmaceutically acceptable salt thereof.

[0081] E19

[0082] [Chemical formula] A compound of Embodiment E1, or a pharmaceutically acceptable salt thereof, selected from the group consisting of

[0083] E20

[0084] [Chemical formula] A compound of Embodiment E1, or a pharmaceutically acceptable salt thereof, selected from the group consisting of

[0085] E21

[0086] [Chemical formula] A compound of Embodiment E1, or a pharmaceutically acceptable salt thereof, selected from the group consisting of

[0087] E21a

[0088] [Chemical formula] A compound of Embodiment E1, or a pharmaceutically acceptable salt thereof, selected from the group consisting of

[0089] E21b

[0090] [Chemical formula] A compound of Embodiment E1, or a pharmaceutically acceptable salt thereof, selected from the group consisting of

[0091] E21c

[0092] [ka] A compound of Embodiment E1, or a pharmaceutically acceptable salt thereof, selected from the group consisting of the following.

[0093] E21d

[0094] [ka] A compound of Embodiment E1, or a pharmaceutically acceptable salt thereof, selected from the group consisting of the following.

[0095] E21e

[0096] [ka] A compound of Embodiment E1, or a pharmaceutically acceptable salt thereof, selected from the group consisting of the following.

[0097] E22

[0098] [ka] A compound, or a pharmaceutically acceptable salt thereof.

[0099] E23

[0100] [ka] It is a compound.

[0101] E24

[0102] [ka] A pharmaceutically acceptable salt of the compound.

[0103] E25

[0104] [Chemical formula] A compound that is or a pharmaceutically acceptable salt thereof.

[0105] E26

[0106] [Chemical formula] A compound that is.

[0107] E27

[0108] [Chemical formula] A pharmaceutically acceptable salt of a compound that is.

[0109] E28

[0110] [Chemical formula] A compound that is or a pharmaceutically acceptable salt thereof.

[0111] E29

[0112] [Chemical formula] A compound that is.

[0113] E30

[0114] [Chemical formula] A pharmaceutically acceptable salt of a compound that is.

[0115] E31

[0116] [Chemical formula] A compound that is or a pharmaceutically acceptable salt thereof.

[0117] E32

[0118] [ka] It is a compound.

[0119] E33

[0120] [ka] A pharmaceutically acceptable salt of the compound.

[0121] E34

[0122] [ka] A compound, or a pharmaceutically acceptable salt thereof.

[0123] E35

[0124] [ka] It is a compound.

[0125] E36

[0126] [ka] A pharmaceutically acceptable salt of the compound.

[0127] E37

[0128] [ka] A compound, or a pharmaceutically acceptable salt thereof.

[0129] E38

[0130] [ka] It is a compound.

[0131] E39

[0132] [ka] A pharmaceutically acceptable salt of the compound.

[0133] E40

[0134] [ka] A compound, or a pharmaceutically acceptable salt thereof.

[0135] E41

[0136] [ka] It is a compound.

[0137] E42

[0138] [ka] A pharmaceutically acceptable salt of the compound.

[0139] E42a

[0140] [ka] A compound, or a pharmaceutically acceptable salt thereof.

[0141] E42b

[0142] [ka] It is a compound.

[0143] E42c

[0144] [ka] A pharmaceutically acceptable salt of the compound.

[0145] E42d

[0146] [ka] A compound, or a pharmaceutically acceptable salt thereof.

[0147] E42e

[0148] [ka] It is a compound.

[0149] E42f

[0150] [ka] A pharmaceutically acceptable salt of the compound.

[0151] E42g

[0152] [ka] A compound, or a pharmaceutically acceptable salt thereof.

[0153] E42h

[0154] [ka] It is a compound.

[0155] E42i

[0156] [ka] A pharmaceutically acceptable salt of the compound.

[0157] E42j

[0158] [ka] A compound, or a pharmaceutically acceptable salt thereof.

[0159] E42k

[0160] [ka] It is a compound.

[0161] E42l

[0162] [ka] A pharmaceutically acceptable salt of the compound.

[0163] E42m

[0164] [ka] A compound, or a pharmaceutically acceptable salt thereof.

[0165] E42n

[0166] [ka] It is a compound.

[0167] E42o

[0168] [ka] A pharmaceutically acceptable salt of the compound.

[0169] E42p

[0170] [ka] A compound, or a pharmaceutically acceptable salt thereof.

[0171] E42q

[0172] [ka] It is a compound.

[0173] E42r

[0174] [ka] A pharmaceutically acceptable salt of the compound.

[0175] E42s

[0176] [ka] A compound, or a pharmaceutically acceptable salt thereof.

[0177] E42t

[0178] [ka] It is a compound.

[0179] E42u

[0180] [ka] A pharmaceutically acceptable salt of the compound.

[0181] E42v

[0182] [ka] A compound, or a pharmaceutically acceptable salt thereof.

[0183] E42w

[0184] [ka] It is a compound.

[0185] E42x

[0186] [ka] A pharmaceutically acceptable salt of the compound.

[0187] E42y

[0188] [ka] A compound, or a pharmaceutically acceptable salt thereof.

[0189] E42a1

[0190] [ka] It is a compound.

[0191] E42a2

[0192] [ka] A pharmaceutically acceptable salt of the compound.

[0193] E43 A pharmaceutical composition comprising one compound from any one of embodiments E1 to E42, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable additive.

[0194] E44 A method for treating cancer, comprising administering a therapeutically effective dose of one of the compounds from embodiments E1 to E42, or a pharmaceutically acceptable salt thereof, to a subject in need thereof.

[0195] E45 A method for treating cancer, comprising administering a therapeutically effective dose of one of the compounds from embodiments E1 to E42, or a pharmaceutically acceptable salt thereof, as a single agent to a subject in need thereof.

[0196] E46 A method for treating cancer, comprising administering a therapeutically effective amount of one of embodiments E1 to E42, or a pharmaceutically acceptable salt thereof, to a subject in need thereof, and further comprising administering a therapeutically effective amount of an additional anticancer drug.

[0197] E47 A method for treating any one of the cancers of Embodiments E22 to E42, wherein the cancer is small cell lung cancer (NSCLC), pancreatic cancer, or colorectal cancer.

[0198] E48 A compound from any one of embodiments E1 to E42, or a pharmaceutically acceptable salt thereof, for use as a pharmaceutical.

[0199] E49 A compound from any one of embodiments E1 to E42, or a pharmaceutically acceptable salt thereof, for use in the treatment of cancer.

[0200] E50 A compound or a pharmaceutically acceptable salt thereof for use in the treatment of cancer according to any one of Embodiments E22 to E42, wherein the cancer is small cell lung cancer (NSCLC), pancreatic cancer, or colorectal cancer.

[0201] E51 Use of any one compound from Embodiments E1 to E42, or a pharmaceutically acceptable salt thereof, for the manufacture of a pharmaceutical for treating cancer.

[0202] E52 Use of a compound, or a pharmaceutically acceptable salt thereof, for manufacturing a medicament for treating cancer according to any one of Embodiments E22 to E42, wherein the cancer is small cell lung cancer (NSCLC), pancreatic cancer, or colorectal cancer.

[0203] E53 A method for treating a disorder mediated by inhibition of KRAS G12C, KRAS G12D, and KRAS G12V receptors in a subject, comprising administering to a subject in need of such treatment one of the compounds from Embodiments E1 to E42, or a pharmaceutically acceptable salt thereof, in an amount effective for treating the disorder.

[0204] E54 A pharmaceutical combination comprising one compound from any of embodiments E1 to E42, or a pharmaceutically acceptable salt thereof, and at least one additional therapeutic agent or a pharmaceutically acceptable salt thereof, wherein the combination is fixed or unfixed.

[0205] E55 A pharmaceutical composition comprising the pharmaceutical combination of Embodiment E54 and at least one additive.

[0206] Each of the embodiments described herein can be combined with any other embodiment described herein that is not inconsistent with the combined embodiment. In addition, any of the compounds described in the examples, or a pharmaceutically acceptable salt thereof, may be claimed individually, or as a group with one or more other compounds from the examples, or a pharmaceutically acceptable salt thereof.

[0207] Furthermore, each of the embodiments described herein intends, within its scope, to be a pharmaceutically acceptable salt, stereoisomer, hydrate, and pharmaceutically acceptable salt of the compound described herein.

[0208] definition Unless otherwise defined herein, scientific and technical terms used in connection with the present invention have meanings commonly understood by those skilled in the art.

[0209] The present invention as described herein can be adequately carried out even in the absence of any element not specifically disclosed herein.

[0210] The "compounds of the present invention" include compounds of formula (I), (II), or (III) and novel intermediates used in their preparation. Those skilled in the art will recognize that the compounds of the present invention include, where possible, conformational isomers (e.g., cis and trans isomers) and all optical isomers (e.g., enantiomers and diastereoisomers), racemates of such isomers, diastereoisomers and other mixtures, and their tautomers. Those skilled in the art will also recognize that the compounds of the present invention include, where possible, their solvates, hydrates, isomorphs, polymorphs, esters, salt forms, prodrugs, and isotopically labeled versions.

[0211] As used herein, the singular forms "a," "an," and "the" also include plural references unless otherwise indicated. For example, "a" substituents include one or more substituents.

[0212] As used herein, the term “about” means that, when used to modify a numerically defined parameter (e.g., a dose of 5 mg), the parameter may vary by 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.

[0213] When it is stated that substituents are "independently selected" from the group, each substituent is selected independently of the others. Therefore, each substituent may be identical or different from the others.

[0214] "Optional" or "optional" means that the event or situation described later may or may not occur, and that the description includes both instances in which the event or situation occurs and instances in which it does not.

[0215] The terms “may be substituted” and “substituted or unsubstituted” are used interchangeably to indicate that a particular group described may not have non-hydrogen substituents (i.e., is unsubstituted) or that the group may have one or more non-hydrogen substituents (i.e., is substituted). Unless otherwise specified, the total number of possible substituents is equal to the number of H atoms present on the unsubstituted form of the group described. If an optional substituent, such as an oxo (=O) substituent, is bonded via a double bond, the group occupies two available valencies, and therefore the total number of other substituents involved is reduced by two. If the optional substituents are selected independently from a list of options, the selected groups may be the same or different. Throughout this disclosure, it will be understood that the number and nature of optional substituents will be limited insofar as such substitutions have chemical significance to those skilled in the art. For clarity, “may be substituted” means substituted with zero or more substituents.

[0216] "Halogens" refer to fluoro, chloro, bromo, and iodine (F, Cl, Br, I).

[0217] "Cyano" refers to a substituent having a carbon atom bonded to a nitrogen atom by a triple bond, i.e., -C≡N (also denoted as "-CN" in this specification).

[0218] "Hydroxy" refers to the -OH group.

[0219] "Oxo" refers to oxygen (=O) that is double-bonded.

[0220] "Alkyl" refers to a saturated monovalent aliphatic hydrocarbon radical having a specified number of carbon atoms, including a linear or branched group. 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 similar groups.

[0221] "Fluoroalkyl" refers to an alkyl 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, fluoromethyl, difluoromethyl, fluoroethyl, difluoroethyl, trifluoroethyl, and tetrafluoroethyl. Examples of completely substituted fluoroalkyl groups (also called perfluoroalkyl groups) include trifluoromethyl (-CF3) and pentafluoroethyl (-C2F5).

[0222] "Alkylene" refers to a divalent aliphatic hydrocarbon radical having a specified number of carbon atoms. An alkylene group may contain, but is 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).

[0223] "Alkoxy" refers to an alkyl group, as defined herein, that is single-bonded to an oxygen atom. The bonding site of the alkoxy radical to the molecule is via the oxygen atom. The 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"). Alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, and similar groups.

[0224] "Alkynyl" refers to an alkyl group as defined herein, comprising at least two carbon atoms and at least one carbon-carbon triple bond. Alkynnyl may contain two to three carbon atoms ("C2-C3 alkynyl"). Examples include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, and similar groups.

[0225] "Cycloalkyl" refers to a fully saturated hydrocarbon ring system having a specified number of carbon atoms, which may be a monocyclic bridge, condensed bicyclic, or polycyclic ring system linked to the base molecule via the carbon atoms of the cycloalkyl ring. Cycloalkyls are not limited to those having 3 to 10 carbon atoms ("C3-C3"). 10 They may contain cycloalkyl groups ("C3-C8 cycloalkyl"), 3-8 carbon atoms ("C3-C8 cycloalkyl"), 3-6 carbon atoms ("C3-C6 cycloalkyl"), 3-5 carbon atoms ("C3-C5 cycloalkyl"), or 3-4 carbon atoms ("C3-C4 cycloalkyl"). Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantanyl, and similar groups. The cycloalkyl groups may be substituted, unsubstituted, or otherwise substituted, as further defined herein.

[0226] "Alkylidennyl" refers to a linear or branched monovalent hydrocarbon radical having the formula =CR'R'', where R' and R'' may be independently selected from H or alkyl groups. Exemplary alkyllidennyl radicals include, but are not limited to, methylidenyl (=CH2), ethylidenyl (=CHCH3), isopropyridenyl (=C(CH3)2), and propyridenyl (=CH-CH2-CH3). R' and R'' may be further substituted as described above.

[0227] A "haloalkylidenyl" refers to a linear or branched monovalent hydrocarbon radical having the formula =CR'R'', where R' or R'' is as defined for alkylidenyl and further contains at least one halogen atom. Exemplary haloalkylidenyl radicals include, but are not limited to, fluoromethylidenyl (=CHF), difluoromethylidenyl (=CF2), fluoroethylidenyl (=CFCH3), and fluoropropyridenyl (=CF-CH2-CH3). R' and R'' may be further substituted as described above.

[0228] "Alkylidenylcyclopropyl" has the formula =CR'R'', where R' or R'' is as defined for alkylidenyl, and R' and R'', together with the carbons to which they are bonded, refer to linear or branched monovalent hydrocarbon radicals that constitute a cyclopropyl ring.

[0229] "Alkylidennyl oxetane" has the formula =CR'R'', where R' or R'' is as defined for alkylidennyl, and R' and R'', together with the carbons to which they are bonded, refer to the linear or branched monovalent hydrocarbon radicals that make up the oxetane.

[0230] "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 fluoro atoms. Examples include, but are not limited to, fluorocyclopropyl, fluorocyclobutyl, fluorocyclopentyl, and fluorocyclohexyl.

[0231] A "heterocycloalkyl" contains a specified number of ring atoms, and contains at least one heteroatom selected from N, O, and S as a ring member, and the ring S atom may be substituted with one or two oxo groups (i.e., S(O) q A heterocycloalkyl ring (where q is 0, 1, or 2) refers to a fully saturated ring system in which a heterocycloalkyl ring is linked to a base molecule via a ring atom that may be C or N. Heterocycloalkyl rings include monocyclic or polycyclic rings, such as bicyclic rings. Heterocycloalkyl rings also include spirocyclic rings, bridged rings, and / or fused rings to one or more other heterocycloalkyl or carbocyclic rings, and such spirocyclic, bridged, and / or fused rings themselves may be saturated, unsaturated, partially unsaturated to the extent that aromaticity has chemical significance, or aromatic, provided that the bonding site to the base molecule is an atom of the heterocycloalkyl portion of the ring system. Heterocycloalkyl rings have N, O, and S(O) as ring members. q It may contain 1 to 4 heteroatoms, or 1 to 3 ring heteroatoms, or 1 to 2 ring heteroatoms selected from the above, provided that such heterocycloalkyl rings do not contain two adjacent oxygen or sulfur atoms.

[0232] The heterocycloalkyl ring may be substituted, unsubstituted, or otherwise substituted, as further defined herein. Such substituents may be located on a heterocyclic ring bonded to the base molecule, or on a monocyclic, bicyclic, tricyclic, spirocyclic, bridging, or fused ring bonded thereto.

[0233] 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 definition herein. Examples of heterocycloalkyl ring groups of the present invention may include, but is not limited to, azilidinyl, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, azepanyl, oxazepanyl, thieazepanyl, a radical of a hexahydro-1H-pyrrolidine ring, a radical of an 8-oxa-3-azabicyclo[3.2.1]octane ring, a radical of a 3-azabicyclo[3.2.1]octane ring, a radical of a 6-azabicyclo[3.2.1]octane ring, or a radical of a 3-azabicyclo[3.2.0]heptane ring.

[0234] "Aryl" or "aromatic" means that it contains a specified number of ring atoms, and all carbon atoms in the ring are sp 2 This refers to a hybridized, pi-conjugated, monocyclic, bicyclic (e.g., biaryl, condensed), or polycyclic ring system. The aryl group is not limited to this, but consists of 6 to 10 carbon atoms ("C6~C"). 10 The fused aryl group may contain an aryl ring (e.g., a phenyl ring) fused to another aryl ring. Examples include, but are not limited to, phenyl, biphenyl, naphthyl, anthracenyl, phenantrenyl, indanyl, and indenyl. The aryl group may be substituted, unsubstituted, or otherwise substituted, as further defined herein.

[0235] Similarly, "heteroaryl" or "heteroaromatic" contains a specified number of ring atoms, with at least one heteroatom selected from N, O, and S as a ring member in the ring, and all carbon atoms in the ring being sp 2This refers to a hybridized, pi-conjugated, monocyclic, bicyclic (e.g., heterobiaryl, condensed), or polycyclic ring system. 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 bonded to the base molecule via the ring atoms of the heteroaromatic ring. Therefore, either a 5- or 6-membered heteroaryl ring may be bonded to the base molecule alone or in a condensed structure via ring C or N atoms.Examples of heteroaryl groups include, but are not limited to, pyrrolyl, furanil, thiophenyl, pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, isothiazolyl, thiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, pyridinyl, pyridizinyl, pyrimidinyl, pyrazinyl, benzofuranil, benzothiophenyl, indolyl, benzimidazolyl, indazolyl, benzotriazolyl, pyrrolo[2,3-b]pyridinyl, and pyrrolo[2,3-c]pyridinyl. Ridinyl, pyrrolo[3,2-c]pyridinyl, pyrrolo[3,2-b]pyridinyl, quinolinyl, isoquinolinyl, purinyl, triazinyl, naphthilidinyl, 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, indolininyl , imidazo[1,2-a]pyridinyl, imidazo[1,5-a]pyridinyl, pyrazolo[1,5-a]pyridinyl, pyrolo[1,2-b]pyridadinyl, imidazo[1,2-c]pyridinyl, azaquinazolinyl, phthalazinyl, (pyrido[3,2-d]pyridinyl, (pyrido[4,3-d]pyridinyl, (pyrido[3,4-d]pyridinyl, (pyrido[2,3-d]pyridinyl, pyrido[2,3-b]pyridinyl, pyrido[3,4-b]pyridinyl, pyrido[5,4-d]pyridinyl, pyrazino[2, This includes [3-b]pyrazinyl and 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. Heteroaryl groups may be substituted, unsubstituted, or otherwise substituted, as further defined herein.

[0236] "Amino" refers to an unsubstituted -NH2 group. Where it is stated that amino is substituted or may be substituted, the term includes groups of the form -NRxRy, where Rx and Ry are defined as further 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 a specified number of carbon atoms (e.g., -NH(C1~C4 alkyl) or -N(C1~C4 alkyl)2).

[0237] In this disclosure, the wavy line used in the chemical structure is "

[0238] [ka] " refers to the bonding point of the substituent.

[0239] The term "pharmaceutically acceptable" means that a substance (e.g., a compound described herein) or any salt thereof, or a composition containing the substance or salt of the present invention, is suitable for administration to a subject or patient.

[0240] As used herein, the term "deuterium enrichment factor" refers to the ratio of the amount of deuterium to the amount of hydrogen and the natural abundance of deuterium, respectively. Atomic arrangements indicated as having deuterium typically have, in certain embodiments, deuterium enrichment factors of at least 1000 (15% deuterium), at least 2000 (30% deuterium), at least 3000 (45% deuterium), at least 3500 (52.5% deuterium), at least 3500 (52.5% deuterium with each indicated deuterium atom), at least 4000 (60% deuterium), at least 4500 (67.5% deuterium), at least 5000 (75% deuterium), at least 5500 (82.5% deuterium), at least 6000 (90% deuterium), at least 6333.3 (95% deuterium), at least 6466.7 (97% deuterium), at least 6600 (99% deuterium), or at least 6633.3 (99.5% deuterium).

[0241] salt The term "pharmaceutically acceptable salt" generally refers to a salt of the present invention prepared by reacting a free base or a free acid with a suitable organic or inorganic acid, or a suitable organic or inorganic base, respectively, to obtain a salt of the present invention that is suitable for administration to a subject or patient.

[0242] In addition, the compounds of formulas (I) to (III) 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) to prepare the compound of formula (I); 2) to purify the compound of formula (I); 3) to separate the enantiomers of the compound of formula (I); or 4) to separate the diastereoisomers of the compound of formula (I).

[0243] Suitable acid addition salts are formed from acids that form non-toxic salts. Examples include, but are not limited to, acetates, adipines, aspartates, benzoates, besilates, bicarbonates / carbonates, bisulfates / sulfates, borates, cansilates, citrates, cyclamates, edisylates, esylates, formates, fumarates, gluceptates, glucons, glucurons, hexafluorophosphates, hibenzates, hydrochlorides / chlorides, hydrobromites / bromids, hydroiodides / iodides, isethionates, This includes lactates, malates, maleates, malons, mesylates, methylsulfates, naphthylates, 2-napsylates, nicotinates, nitrates, orotates, oxalates, palmitates, pamoates, phosphates / hydrogen phosphates / dihydrogen phosphates, pyroglutamates, saccharates, stearates, succinates, tannates, tartrates, tosylates, trifluoroacetates, 1,5-naphthalenedisulfonic acid, and xinofoates.

[0244] 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.

[0245] Hemisalts of acids and bases, such as hemisulfates and hemicalcium salts, may also be formed.

[0246] For an overview of suitable salts, see Paulekun, GS 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.

[0247] A pharmaceutically acceptable salt of the compound of the present invention can be prepared by methods well known to those skilled in the art, including, but not limited to, the following procedure. (i) A procedure involving reacting the compound of the present invention with a desired acid or base; (ii) a procedure by removing an acid or base instability protecting group from a suitable precursor of the compound of the present invention, or by opening the ring of a suitable cyclic precursor, such as a lactone or lactam, using a desired acid or base; or (iii) A procedure for converting one salt of the compound of the present invention to another salt. This can be achieved by reacting it with a suitable acid or salt, or by using a suitable ion exchange procedure.

[0248] These procedures are typically carried out in solution. The resulting salt can be precipitated and filtered off, or recovered by evaporation of the solvent.

[0249] solvate The compounds of the present invention and their pharmaceutically acceptable salts may exist in non-solvated and solvated forms. The term “solvate” is used herein to describe a molecular complex comprising the compound of the present invention or its pharmaceutically acceptable salt with one or more pharmaceutically acceptable solvent molecules, such as ethanol. The term “hydrate” is used when the solvent is water.

[0250] In addition, the compounds of formulas (I) to (III) may also 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) to prepare the compounds of formulas (I) to (III); 2) to purify the compounds of formulas (I) to (III); 3) to separate the enantiomers of the compound of formula (I); or 4) to separate the diastereoisomers of the compounds of formulas (I) to (III).

[0251] The currently accepted classification system for organic hydrates defines them as isolated site, channel, or metal ion-coordinated hydrates. See KRMorris's *Polymorphism in Pharmaceutical Solids* (HGBrittain, Marcel Dekker, 1995). Isolated site hydrates are hydrates in which water molecules are isolated from direct contact with each other by the interposition of organic molecules. In channel hydrates, water molecules exist within lattice channels, where they are adjacent to other water molecules. In metal ion-coordinated hydrates, water molecules are bound to metal ions.

[0252] When the solvent or water is tightly bound, the complex can have a well-defined stoichiometry, independent of humidity. However, when the solvent or water is weakly bound, as in channel solvates and hygroscopic compounds, the water / solvent content can depend on humidity and dry conditions. In such cases, non-stoichiometry is the norm.

[0253] complex Multicomponent complexes (other than salts and solvates) in which a drug and at least one other component exist 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 cocrystals. The latter are typically defined as crystalline complexes of neutral molecular components bonded together by non-covalent interactions; for example, hydrogen-bonded complexes (cocrystals) can be formed with neutral molecules or salts. Cocrystals can be prepared by melt crystallization, recrystallization from a solvent, or physical grinding of the components. See Chem Commun, 17;1889–1896 by O. Almarsson and MJ Zaworotko (2004). For a general overview of multicomponent complexes, see J Pharm Sci, 64(8), 1269–1288 by Haleblian (August 1975).

[0254] 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 where 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. When heated, 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 where 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").

[0255] 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 the true crystalline state and the true liquid state (either melt or solution), and consists of two-dimensional order at the molecular level. Intermediate states resulting from temperature changes are described as "thermotropic", and those resulting from adding a second component such as water or another solvent are 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).

[0256] Stereoisomers The compounds of the present invention may exist as two or more stereoisomers. Stereoiomers of a compound 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, a compound of the present invention containing one or more chiral carbon atoms may exist as two or more stereoisomers.

[0257] 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).

[0258] The cis / trans isomers can be separated by conventional techniques well known to those skilled in the art, such as chromatography and fractional crystallization.

[0259] Conventional techniques for preparing / isolating individual enantiomers include chiral synthesis from suitable optically pure precursors, or separation of racemates (or racemates of salts or derivatives) using, for example, chiral high-pressure liquid chromatography (HPLC). Alternatively, the racemate (or racemic precursor) can be reacted with a suitable optically active compound, such as an alcohol, or, if the compound of the present invention contains an acidic or basic moiety, with 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. Using chromatography, the chiral compound of the present invention (and its chiral precursor) can be obtained in an enantiomerically concentrated form, and typically, the concentrated mixture is obtained by HPLC concentration of the eluent. Chiral chromatography using subcritical and supercritical fluids can be used. Methods for chiral chromatography useful in some embodiments of the present 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), pp. 223-249 and the references cited therein).

[0260] When any racemic mixture crystallizes, two different types of crystals can occur. The first type is the racemic compound (true racemic compound) mentioned above, which produces a single, uniform crystal form containing equimolar amounts of both enantiomers. The second type is a racemic mixture or aggregate in which equimolar amounts of two crystalline forms, each containing a single enantiomer, are produced. Both crystalline forms present in a racemic mixture have the same physical properties, although they may have different physical properties compared to a true racemic mixture. Racemic mixtures can be separated by prior art known to those skilled in the art. See, for example, Stereochemistry of Organic Compounds by ELEEliel and SHWilen (Wiley, New York, 1994).

[0261] Tautomerism When structural isomers are interconvertible across low-energy barriers, tautomeric isomerism ("tautomerism") can occur. This can take the form of proton tautomerism in the compounds of the present invention containing imino / amino, keto / enol, or oxime / nitroso groups, or lactam / lactim, or so-called valence tautomerism in compounds containing aromatic moieties. This means that a single compound may exhibit more than one type of isomer.

[0262] For the sake of brevity, the compounds of the present invention are illustrated herein in a single tautomer, but it should be emphasized that all possible tautomers are included within the scope of the present invention.

[0263] Isotopes The present invention encompasses all pharmaceutically acceptable isotope-labeled compounds of the present invention, in which one or more atoms are replaced by atoms having the same atomic number but having an atomic mass or mass number different from the atomic mass or mass number that is dominant in nature.

[0264] Examples of isotopes suitable for inclusion in the compound of the present invention include:2 H and 3 hydrogen such as H, 11 C, 13 C and 14 carbon such as C, 36 chlorine such as Cl, 18 fluorine such as F, 123 I and 125 iodine such as I, 13 N and 15 nitrogen such as N, 15 O, 17 O and 18 oxygen such as O, 32 phosphorus such as P, and 35 sulfur isotopes such as S may be included.

[0265] Certain isotope-labeled compounds of the present invention, for example, those incorporating radioisotopes, are useful in one or both of drug or substrate tissue distribution studies. Radioisotope tritium, that is, 3 H, and carbon-14, that is, 14 C are particularly useful for this purpose considering their ease of incorporation and rapid means of detection.

[0266] Deuterium, that is, 2 substitution with H can provide certain therapeutic advantages resulting from greater metabolic stability.

[0267] 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.

[0268] In some embodiments, this disclosure provides deuterium-labeled (or deuterated) compounds and salts, the formulas and variants of such compounds and salts being as described herein, respectively, and independently. “Deuterated” means that at least one of the atoms in the compound is deuterium in an abundance higher than the natural abundance of deuterium (typically about 0.015%). Experienced practitioners have recognized that in chemical compounds containing hydrogen atoms, the hydrogen atoms actually exist as a mixture of H and D, with about 0.015% being D. The concentration of deuterium incorporated into the deuterium-labeled compounds and salts of the present invention may be defined by the deuterium concentration factor. It is understood that one or more deuterium atoms may be exchanged for hydrogen under physiological conditions.

[0269] In some embodiments, the deuterium compound is selected from any one of the compounds described in Table 2 shown in the Examples section.

[0270] In some embodiments, one or more hydrogen atoms on a specific metabolic site on the compound of the present invention are deuterated.

[0271] The isotope-labeled compounds of the present invention can generally be prepared by conventional techniques known to those skilled in the art, or by processes similar to those described in the attached examples and preparation examples, using a suitable isotope-labeled reagent instead of a previously used unlabeled reagent.

[0272] The pharmaceutically acceptable solvates according to the present invention include those in which the crystallization solvent may be isotope-substituted, for example, D2O, d6-acetone, or d6-DMSO.

[0273] Prodrug The compounds of the present invention can be administered in the form of prodrugs. Therefore, certain derivatives of the compounds of the present invention that themselves have little or no pharmacological activity may, when administered intracellularly or onto the body, be converted to compounds of the present invention having desired activity, for example, by hydrolytic cleavage, particularly catalyzed 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.).

[0274] The prodrugs according to the present invention can be produced, for example, by replacing a suitable functional group present in the compound of the present invention with a specific part known to those skilled in the art, as a "pro-mole," as described in "Design of Prodrugs" by H. Bundgaard (Elsevier, 1985).

[0275] Therefore, the prodrug according to the present invention may be (a) an ester or amide derivative of a carboxylic acid when present in the compound of the present invention; (b) an ester, carbonate, carbamate, phosphate or ether derivative of a hydroxyl group when present in the compound of the present invention; (c) an amide, imine, carbamate or amine derivative of an amino group when present in the compound of the present invention; (d) a thioester, thiocarbonate, thiocarbamate or sulfide derivative of a thiol group when present in the compound of the present invention; or (e) an oxime or imine derivative of a carbonyl group when present in the compound of the present invention.

[0276] Some specific examples of 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 C1-C8 alkyl (e.g., ethyl) or (C1-C8 alkyl)C(=O)OCH2- (e.g., t Compounds that are replaced by BuC(=O)OCH2-); (ii) When the compound of the present invention contains an alcohol functional group (-OH), an ester thereof, for example, a compound in which the hydrogen of the alcohol functional group of the compound is replaced by -CO(C1~C8 alkyl) (e.g., methyl carbonyl), or in which the alcohol is esterified with an amino acid; (iii) If the compound of the present invention contains an alcohol functional group (-OH), then an ether thereof, for example, a compound in which 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) If the compound of the present invention contains an alcohol functional group (-OH), then the phosphate, for example, the hydrogen of the alcohol functional group of the compound is -P(=O)(OH)2 or -P(=O)(O - Na + )2 or -P(=O)(O - )2Ca 2+ Compounds that have been replaced by; (v) If the compound of the present invention contains a primary or secondary amino functional group (-NH2 or -NHR, where R≠H), then the amide, for example, optionally, one or both of the hydrogen atoms of the amino functional group of the compound are (C1~C 10 ) Compounds in which the amino group is replaced by an alkanoyl group, -COCH2NH2, or the amino group is derivatized with an amino acid; (vi) If the compound of the present invention contains a primary or secondary amino functional group (-NH2 or -NHR, where R≠H), then the amine, for example, a compound in which one or both hydrogens of the amino functional group of the compound are replaced by -CH2OP(=O)(OH)2. It includes.

[0277] Certain compounds of the present invention can act as prodrugs of other compounds of the present invention. It is also possible for two compounds of the present invention to act together as a prodrug. In certain circumstances, a prodrug of a compound of the present invention can be made by internally linking two functional groups in the compound of the present invention, for example, to form a lactone.

[0278] metabolites The active metabolites of the compounds of the present invention, i.e., compounds that are often formed in vivo by oxidation or dealkylation upon administration of the drug, are also included within the scope of the present invention. Some examples of metabolites according to the present invention, but not limited to these: (i) If the compound of the present invention contains an alkyl group, its hydroxyalkyl derivative (-CH > -COH): (ii) If the compound of the present invention contains an alkoxy group, its hydroxy derivative (-OR → -OH); (iii) If the compound of the present invention contains a tertiary amino group, its secondary amino derivative (-NRR' → -NHR or -NHR'); (iv) If the compound of the present invention contains a secondary amino group, its primary derivative (-NHR → -NH2); (v) If the compound of the present invention contains a phenyl moiety, its phenol derivative (-Ph → -PhOH); (vi) If the compound of the present invention contains an amide group, its carboxylic acid derivative (-CONH2 → COOH); and (vii) If the compound contains a hydroxyl or carboxylic acid group, the compound may be metabolized, for example, by conjugation with glucuronic acid to form a glucuronide. Other conjugation metabolic pathways 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 be conjugated.

[0279] Pharmaceutical composition In another embodiment, the present invention includes pharmaceutical compositions. For the purposes of pharmaceutical compositions, the compound itself or a pharmaceutically acceptable salt thereof will simply be referred to as the compound of the present invention.

[0280] "Pharmaceutical composition" refers to a mixture of one or more of the compound of the present invention as an active ingredient, or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug thereof, and at least one pharmaceutically acceptable additive.

[0281] The term "additive" is used herein to describe any component other than the compounds of the present invention. The selection of an additive will depend largely on factors such as the mode of administration, the effect of the additive on solubility and stability, and the properties of the dosage form.

[0282] As used herein, “additives” include all physiologically compatible solvents, dispersions, coatings, antimicrobial and antifungal agents, isotonic and absorption retardants, carriers, diluents, and the like. Examples of additives include one or more of water, saline, phosphate buffer solution, dextrose, glycerol, ethanol, and the like, or combinations thereof, and may also 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). Pharmaceutical compositions may optionally contain additional additives, such as flavorings, binders / binding agents, lubricants, disintegrants, sweeteners or flavorings, colorants or pigments, and the like. For example, for oral administration, tablets containing various additives such as citric acid can be used with various disintegrants such as starch, alginic acid, and certain complex silicates, and binders such as sucrose, gelatin, and gum arabic. Examples of additives, though not limited to them, include calcium carbonate, calcium phosphate, various types of sugars and starches, 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. Similar types of solid compositions can also be used in filled soft and hard gelatin capsules. Therefore, non-limiting examples of additives also include lactose and high molecular weight polyethylene glycol. When aqueous suspensions or elixirs are preferred for oral administration, the active compounds therein can be combined with various sweeteners or flavorings, colorants or pigments, and, if desired, emulsifiers or suspending agents, along with additional additives such as water, ethanol, propylene glycol, glycerin, or combinations thereof.

[0283] Examples of additives include pharmaceutically acceptable substances that enhance the shelf life or efficacy of a compound, such as humectants or small amounts of auxiliary substances, such as humectants or emulsifiers, preservatives, or buffers.

[0284] The compositions of the present invention may take various forms. These include, for example, liquid formulations (e.g., formulations for injection and infusion), dispersants or suspensions, and liquid, semi-solid, and solid dosage forms such as tablets, capsules, pills, powders, liposomes, and suppositories. The form depends on the intended mode of administration and therapeutic application.

[0285] Typical compositions are in the form of injectable or infusion solutions, such as compositions similar to 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.

[0286] Oral administration in solid dosage forms can be provided 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, the oral administration may be in powder or granular form. In another embodiment, the oral dosage form is sublingual, such as lozenges. In such solid dosage forms, the compounds of the present invention are typically combined with one or more adjuvants. Such capsules or tablets may contain controlled-release formulations. In the case of capsules, tablets, and pills, the dosage forms may contain buffers or may be prepared with enteric coatings.

[0287] In another embodiment, oral administration may be in liquid form. Liquid dosage forms for oral administration include, for example, pharmaceutically acceptable emulsions, liquids, suspensions, syrups, and elixirs containing an inert diluent commonly used in the art (e.g., water). Such compositions may contain auxiliary agents such as one or more wetting agents, emulsifiers, suspending agents, flavoring agents (e.g., sweeteners), or fragrances.

[0288] In another embodiment, the present invention includes parenteral dosage forms. "Pareral administration" includes, for example, subcutaneous injections, intravenous injections, intraperitoneal and intramuscular injections, intrasternal injections, and injectable preparations. Preparations for injection (i.e., sterile aqueous or oily suspensions for injection) can be formulated using one or more suitable dispersants, wetting agents, or suspending agents according to known techniques.

[0289] In another embodiment, the present invention includes topical dosage forms. "Topical administration" includes, for example, transdermal and transdermal administration such as transdermal patches or ion electrophoresis devices, 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 area. When the compounds of the present invention are administered by a transdermal device, administration will be achieved using patches of either reservoir and porous membrane type or solid matrix variants. Typical formulations for this purpose include gels, hydrogels, lotions, liquids, creams, ointments, sprays, bandages, foams, films, skin patches, wafers, implants, sponges, fibers, adhesive bandages, and microemulsions. Liposomes may also be used. Typical additives include alcohol, water, mineral oil, liquid petrolatum, white petrolatum, glycerin, polyethylene glycol, and propylene glycol. A permeation enhancer can be incorporated. For example, see BCFinnin and TMMorgan, J.Pharm.Sci., vol.88, pp. 955-958, 1999.

[0290] Formulations suitable for topical administration to the eye include, for example, eye drops in which the compound of the present invention is dissolved or suspended in a suitable additive. Typical formulations suitable for ocular or ocular administration may be in the form of ultrafine-ground suspensions or solution drops in isotonic, pH-adjusted sterile saline. Other formulations suitable for ocular and ocular administration include ointments, biodegradable (i.e., absorbent gel sponges, collagen) and non-biodegradable (i.e., silicone) implants, wafers, lenses, and microparticles or vesicles such as niosomes or liposomes. Cross-linked polyacrylic acid, polyvinyl alcohol, hyaluronic acid, cellulosic polymers, such as hydroxypropyl methylcellulose, hydroxyethylcellulose, or methylcellulose, or heteropolysaccharide polymers, such as geran gum, can be incorporated together with preservatives such as benzalkonium chloride. Such formulations can also be delivered by ion electrophoresis.

[0291] For intranasal administration, the compounds of the present invention are conveniently delivered in the form of a solution or suspension from a pump-type spray container squeezed or pumped by the patient, or as an aerosol spray dispensing from a pressurized container or nebulizer using a suitable propellant. Formulations suitable for intranasal administration are typically administered in the form of a dry powder from a dry powder inhaler (either alone or as a mixture, e.g., a dry blend with lactose, or as mixed component particles, e.g., mixed with phospholipids such as phosphatidylcholine), or as an aerosol spray from a pressurized container, pump, spray, atomizer (preferably an atomizer using electrohydrodynamics to produce a fine mist), or from a nebulizer with or without a suitable propellant such as 1,1,1,2-tetrafluoroethane or 1,1,1,2,3,3,3-heptafluoropropane. For intranasal use, the powder may contain a bioadhesive, e.g., chitosan or cyclodextrin.

[0292] In another embodiment, the present invention includes a rectal dosage form. Such a rectal dosage form may be, for example, a suppository. Cocoa butter is the conventional suppository base, but various alternatives can be used if appropriate.

[0293] Other additives and modes of administration known in the pharmaceutical field may also be used. The pharmaceutical compositions of the present invention can be prepared by any of the well-known pharmaceutical techniques, such as effective formulations and administration procedures. The above considerations regarding effective formulations and administration procedures are well-known in the art and are described in standard textbooks. The formulations of drugs are 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, NY, 1980; and Kibbe et al., Handbook of Pharmaceutical Excipients (Third Edition), American Pharmaceutical Association, Washington, 1999.

[0294] Acceptable additives are nontoxic to the subject at the dosage and concentration used and may include one or more of the following: 1) buffering agents 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 octadecyldimethylbenzylammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl or benzyl alcohol; 5) alkylparabens such as methyl or propylparaben, catechol, resorcinol, cyclohexanol, 3-pentanol, or m-cresol; 6) low molecular weight (less than approximately 10 residues) polypeptides; 7) serum albumin, 1) Proteins such as gelatin or immunoglobulins; 8) Hydrophilic polymers such as polyvinylpyrrolidone; 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 polysorbates (e.g., polysorbate 20 or polysorbate 80), poloxamer, or polyethylene glycol (PEG).

[0295] For oral administration, the composition may 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 for the patient. The pharmacopoeia typically contains about 0.01 mg to about 500 mg of the active ingredient, or in another embodiment, about 1 mg to about 100 mg of the active ingredient. Intravenously, the dose may range from about 0.01 to about 10 mg / kg / min during constant-rate infusion.

[0296] Liposomes containing the compounds of the present invention can be prepared by methods known in the art (see, for example, Chang, HI; Yeh, MK; 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 reverse-phase evaporation using a lipid composition containing phosphatidylcholine, cholesterol, and PEG-derivativeized phosphatidylethanolamine (PEG-PE). The liposomes are extruded through a filter of a specified pore size to obtain liposomes with a desired diameter.

[0297] The compounds of the present invention can also be encapsulated in microcapsules prepared, for example, by coacervation technology or interfacial polymerization, such as hydroxymethylcellulose or gelatin microcapsules and poly-(methyl methacrylate) microcapsules, using colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules) or macroemulsions, respectively. Such techniques are disclosed in Remington, The Science and Practice of Pharmacy, 20th edition, Mack Publishing (2000).

[0298] Sustained-release preparations can be used. A preferred example of a sustained-release preparation comprises a semipermeable matrix of a solid hydrophobic polymer containing the compound of the present invention, the matrix being in the form of a molded article, e.g., a film, 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 (injectable microspheres consisting of lactic acid-glycolic acid copolymer and leuprolide acetate), such as those used in leuprolide acetate for depot suspensions, sucrose acetate isobutyrate, and poly-D-(-)-3-hydroxybutyric acid.

[0299] Preparations used for intravenous administration must be sterile. This can be easily achieved, for example, by filtration through a sterile filtration membrane. The compounds of the present invention are generally placed in containers having a sterile access port, such as intravenous fluid bags or vials with a stopper that can be pierced by a subcutaneous injection needle.

[0300] 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 phospholipids, 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, by other means, in an emulsion formed by mixing oil (e.g., soybean oil, safflower oil, cottonseed oil, sesame oil, corn oil, or tonsil oil) and phospholipids (e.g., egg phospholipids, soybean phospholipids, or soybean lecithin) with water. It will be found that other components, such as glycerol or glucose, can be added to adjust the tonicity of the emulsion. A suitable emulsion should typically contain up to 20% oil, for example, between 5 and 20%. The lipid emulsion contains lipid droplets between 0.1 and 1.0 μm, particularly between 0.1 and 0.5 μm, and may have a pH in the range of 5.5 to 8.0.

[0301] For example, an emulsion composition may be prepared by mixing the compound of the present invention with a lipid emulsion containing soybean oil or its components (soybean oil, egg phospholipids, glycerol, and water).

[0302] Compositions for inhalation or inhalation include liquid and suspension formulations in pharmaceutically acceptable aqueous or organic solvents, or mixtures thereof, as well as powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable additives as presented above. In some embodiments, compositions are administered orally or via nasal respiratory routes for topical or systemic action. Compositions in preferably sterile, pharmaceutically acceptable solvents can be atomized using gas. The atomized solution can be inhaled directly from the atomizing device, or the atomizing device can be attached to a face mask, tent, or intermittent positive pressure ventilator. Solutions, suspensions, or powder compositions can be administered preferably orally or nasally from a device that delivers the formulation in an appropriate manner.

[0303] A formulation intermediate (DPI) is a partially processed substance that requires further processing steps before becoming a bulk formulation. The compounds of the present invention can be formulated into formulation intermediate DPIs containing the active ingredient in a form with a higher free energy than the crystalline form. One reason for using DPIs is to improve the oral absorption characteristics due to low solubility, slow dissolution, improved material transport via the mucin layer adjacent to epithelial cells, and, in some cases, to overcome limitations due to biological barriers such as metabolism and transporters. Other reasons may include improved solid-state stability and downstream manufacturability. In one embodiment, the formulation intermediate contains the compound of the present invention (e.g., an amorphous solid dispersion (ASD)) isolated and stabilized in an amorphous state. Many techniques exist known in the art to produce ASDs that produce substances suitable for incorporation into bulk formulations, e.g., spray-dried dispersions (SDDs), melt extrudes (often referred to as HMEs), co-precipitates, amorphous drug nanoparticles, and nanoadsorbents. In one embodiment, the amorphous solid dispersion contains the compound of the present invention and polymer additives. The concentrations of other additives, as well as the aforementioned additives and the compounds of the present invention, are well known in the art and are described in standard textbooks. For example, see "Amorphous Solid Dispersions Theory and Practice" by Navnit Shah et al.

[0304] Administration and medication As used herein, the terms “to treat,” “to treat,” or “treatment” encompass both preventive, protective, and palliative treatments, i.e., reducing, mitigating, or slowing the progression of any tissue damage to a patient’s disease (or condition) or disease-related condition.

[0305] As used herein, the terms “subject,” “individual,” or “patient,” used interchangeably, refer to any animal, including mammals. Mammals according to the present invention include dogs, cats, cattle, goats, horses, sheep, pigs, rodents, rabbits, primates, humans, and similar animals, and encompass mammals in utero. In one embodiment, humans are preferred subjects. Human subjects may be of any sex and at any developmental stage.

[0306] As used herein, the term “therapeutic dose” means the amount of an active compound or medicinal substance 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, including one or more of the following: (1) To prevent disease; for example, to prevent disease, pathology, or disorder in individuals who are susceptible to disease, pathology, or disorder but have not yet experienced or presented pathology or overall symptoms; (2) inhibiting the disease; for example, inhibiting the disease, condition, or disorder in an individual experiencing or presenting the pathology or overall symptoms of the disease, condition, or disorder (i.e., preventing (or slowing) the further development of the pathology or overall symptoms or both); and (3) Remission of a disease; for example, in an individual experiencing or presenting with the pathology or overall symptoms of a disease, condition, or disorder, remission of the disease, condition, or disorder (i.e., reversal of the pathology or overall symptoms or both).

[0307] Typically, the compounds of the present invention are administered in amounts effective to treat the conditions described herein. The compounds of the present invention may be administered as the compounds themselves, or, by other means, as pharmaceutically acceptable salts. For the purposes of administration and drug delivery, the compounds themselves or their pharmaceutically acceptable salts shall simply be referred to as the compounds of the present invention.

[0308] The compounds of the present invention are administered by any preferred route, in the form of a pharmaceutical composition adapted to that route, and in a dose effective for the intended treatment. The compounds of the present invention can be administered orally, rectally, vaginally, parenterally, topically, intranasally, or by inhalation.

[0309] The compounds of the present invention can be administered orally. Oral administration may involve swallowing, which allows the compound to enter the gastrointestinal tract, or it may be administered buccally or sublingually, allowing the compound to enter the bloodstream directly from the mouth.

[0310] In another embodiment, the compounds of the present invention may be administered parenterally, for example, directly into the bloodstream, muscle, or internal organs. Suitable means for parenteral administration include intravenous, intra-arterial, intraperitoneal, intrathecal, intraventricular, intraurethral, ​​intrasternal, intracranial, intramuscular, and subcutaneous. Suitable devices for parenteral administration include needle (including microneedle) syringes, needleless syringes, and infusion techniques.

[0311] In another embodiment, the compounds of the present invention may be administered topically to the skin or mucous membrane, i.e., cutaneously or transdermally. In another embodiment, the compounds of the present invention may be administered intranasally or by inhalation. In another embodiment, the compounds of the present invention may be administered rectally or vaginally. In another embodiment, the compounds of the present invention may be administered directly to the eyes or ears.

[0312] The administration regimen for the compound of the present invention or a composition containing the compound is based on various factors, including the patient's species, age, weight, sex, and medical condition; the severity of the condition; the route of administration; and the activity of the specific compound used. Therefore, the administration regimen can vary widely. In one embodiment, the total daily dose of the compound 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 indications discussed herein. In another embodiment, the total daily dose of the compound of the present invention is about 0.1 to about 50 mg / kg, and in yet another embodiment, about 0.5 to about 30 mg / kg. It is not uncommon to repeat the administration of the compound of the present invention multiple times a day (typically no more than four times). If desired, multiple doses per day can typically be used to increase the total daily dose.

[0313] Treatment methods and use The compounds of the present invention can inhibit the activity of one or more of the KRAS G12C, KRAS G12D, and KRAS G12V receptors and may be useful in the treatment, prevention, suppression, and remission of diseases such as cancer, disorders, and pathological conditions mediated by any of the KRAS G12C, KRAS G12D, and KRAS G12V receptors, or combinations thereof.

[0314] Cancers 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 non-muscle-invasive bladder cancer, hepatocellular carcinoma, breast cancer, and head and neck cancer.

[0315] Preferably, the compounds of the present invention may be useful for treating lung cancers such as non-small cell lung cancer (NSCLC), pancreatic cancer, colorectal cancer, breast cancer, hematological cancers, gynecological cancers, prostate cancer, or skin cancer. See Mustachio, L., Targeting KRAS in Cancer: Promising Therapeutic Strategies, Cancers, 2021, 13, 1204.

[0316] More preferably, the compounds of the present invention may be useful for treating non-small cell lung cancer (NSCLC), pancreatic cancer, and colorectal cancer.

[0317] 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 use, method or composition as defined herein, of using the compounds of the present invention, or pharmaceutically acceptable salts thereof, in combination with one or more other therapeutic anticancer drugs discussed herein.

[0318] The administration of two or more compounds "in combination" means that all compounds are administered in sufficient time proximity to affect the treatment in question. Two or more compounds may be administered synchronously or sequentially, with or without specific time intervals, via the same or different routes of administration, on the same or different administration schedules, depending on the treatment regimen. In addition, synchronous administration can be achieved by mixing the compounds before administration, or by administering the compounds at the same time, but at the same or different administration sites, in separate dosage forms. Examples of "in combination" include, but are not limited to, "simultaneous administration," "co-administration," "synchronous administration," "sequential administration," and "administered synchronously."

[0319] The compounds of the present invention and one or more other therapeutic agents can be administered as fixed or unfixed combinations 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 "unfixed 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 so that they can be administered synchronously to a subject in need, or at different times with a variable intervening time that allows such administration to bring effective levels of two or more compounds in the subject's body.

[0320] 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-inhibiting plant alkaloids, cytotoxic / antitemonic antibiotics, topoisomerase inhibitors, photosensitizers, anti-estrogen and selective estrogen receptor modulators (SERMs), anti-progesterones, estrogen receptor downregulators (ERDs), estrogen receptor antagonists, luteinizing hormone-releasing hormone agonists; IL-2 receptor agonists (agonists for recombinant cytokines or cytokine receptors); and antisense oligonucleotides or oligonucleotide derivatives that inhibit the expression of genes involved in abnormal cell proliferation or tumor growth.

[0321] Other additional chemotherapy drugs include not only taxanes or platinum-based drugs, but also HER2-targeted drugs, such as trastuzumab.

[0322] In another embodiment, such additional anticancer 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 gedatricib); 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, bioresponse modifiers; enzyme inhibitors; and cytotoxic agents.

[0323] In another embodiment, such additional anticancer agents include, for example, anti-angiogenic agents including tyrosine kinase / vascular endothelial growth factor (VEGF) receptor (VEGFR) inhibitors (including sunitinib, axitinib, sorafenib, and tivozanib), TIE-2 inhibitors, PDGFR inhibitors, angiopoietin inhibitors, PKCβ inhibitors, COX-2 (cyclooxygenase II) inhibitors, integrins (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®), bevacizumab (Avastin®), axitinib (Inlyta®), SU14813 (Pfizer), and AG13958 (Pfizer). Additional anti-angiogenic drugs include batalanib (CGP79787), pegaptanib octasodium (Macugen®), vandetanib (Zactima®), PF-0337210 (Pfizer), SU14843 (Pfizer), AZD2171 (AstraZeneca), ranibizumab (Lucentis®), Neovastat® (AE941), tetrathiomolybdata (Coprexa®), AMG706 (Amgen), and VEGF. These include Trap (AVE0005), CEP7055 (Sanofi-Aventis), XL880 (Exelixis), Teratinib (BAY57-9352), and CP-868,596 (Pfizer). Other anti-angiogenic drugs include Enzastaurin (LY317615), Midostaurin (CGP41251), Perifosine (KRX0401), Teprenone (Selbex®), and UCN 01 (Kyowa Hakko).Other examples of anti-angiogenic drugs include celecoxib (Celebrex®), parecoxib (Dynastat®), delacoxib (SC59046), lumiracoxib (Preige®), valdecoxib (Bextra®), rofecoxib (Vioxx®), iguratimod (Careram®), IP751 (Invedus), SC-58125 (Pharmacia), and etoricoxib (Arcoxia®). Further anti-angiogenic drugs include exisrind (Aptosyn®), sarsalat (Amigesic®), diflunisal (Dolobid®), ibuprofen (Motrin®), ketoprofen (Orudis®), nabumetone (Relafen®), piroxicam (Feldene®), naproxen (Aleve®, Naprosyn®), diclofenac (Voltaren®), indomethacin (Indocin®), sulindac (Clinoril®), tolmetin (Tolectin®), etodolac (Lodine®), ketrolac (Toradol®), and oxaprozin (Daypro®). Further anti-angiogenic drugs include ABT510 (Abbott), aplatastat (TMI005), AZD8955 (AstraZeneca), incyclinide (Metastat®), and PCK3145 (Procyon).Further anti-angiogenic drugs include acitretin (Neotigason®), pritisin (aplidine®), silengutide (EMD121974), combretastatin A4 (CA4P), fenretinide (4HPR), halofuginone (Tempostatin®), Panzem® (2-methoxyestradiol), and PF-03446962 (Pfi This includes zer), Levimast (BMS275291), Katsumakisomab (Removab®), Lenalidomide (Revlimid®), Squalamine (EVIZON®), Thalidomide (Thalomid®), Ukrain® (NSC631570), Vitaxin® (MEDI522), and Zoledronic acid (Zometa®).

[0324] In another embodiment, such additional anticancer agents include compounds derived from hormonal agents and antagonists. Examples include anti-estrogens and selective estrogen receptor modulators (SERMs), as well as selective estrogen receptor degraders (SERDs), where the anti-hormonal agent acts to modulate or inhibit the hormonal action on the tumor, such as tamoxifen, raloxifene, droloxifene, 4-hydroxytamoxifen, trioxyfen, keoxyfen, LY117018, onapristone, toremifene (Fareston), and fulvestrant. Examples include aromatase inhibitors that inhibit aromatase, an enzyme that regulates estrogen production in the adrenal gland, such as 4(5)-imidazole, aminoglutethimide, megestrol acetate, exemestane, formestan, fadrozol, borozole, letrozole, and anastrozole; as well as compounds like antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide, flurisyl, apalutamide, enzalutamide, cimetidine, and goserelin.

[0325] In another embodiment, such additional anticancer therapeutics include signaling inhibitors, such as protein tyrosine kinase and / or serine / threonine kinase inhibitors: compounds derived from signaling inhibitors (e.g., inhibiting the means by which regulatory molecules thereby control fundamental processes of cell proliferation, differentiation, and survival transmitted within a cell). Signaling inhibitors include small molecules, antibodies, and antisense molecules. Signaling inhibitors include, for example, kinase inhibitors (e.g., tyrosine kinase inhibitors or serine / threonine kinase inhibitors) and cell cycle inhibitors. More specifically, signaling inhibitors include, for example, farnesyl protein transferase inhibitors, EGF inhibitors, ErbB-1 (EGFR), ErbB-2, pan erb, IGF1R inhibitors, MEK (including binimetinib (Mektovi®)), c-Kit inhibitors, FLT-3 inhibitors, K-Ras inhibitors, PI3 kinase inhibitors, JAK inhibitors, STAT inhibitors, Raf kinase inhibitors, BRAF (including encorafenib (Braftovi®)), Akt inhibitors, mTOR inhibitors, P70S6 kinase inhibitors, WNT pathway inhibitors, and multitarget kinase inhibitors.

[0326] In another embodiment, such additional anticancer drugs include docetaxel, paclitaxel, paclitaxel protein-binding particles, cisplatin, carboplatin, oxaliplatin, capecitabine, gemcitabine, or vinorelbine.

[0327] In another embodiment, such additional anticancer agents include compounds derived from epigenetic modulators, in which case 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.

[0328] In another embodiment, such additional anticancer agents include compounds that are cancer immunotherapies, including immunomodulators.

[0329] In another embodiment, a combination with pattern recognition receptors (PRRs) is intended. 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 target. There are several groups of PRR molecules, including Toll-like receptors (TLRs), RIG-I-like receptors (RLRs), nucleotide-binding oligomerized domain (NOD)-like receptors (NLRs), C-type lectin receptors (CLRs), and interferon gene-stimulating (STING) proteins.

[0330] STING proteins function as both cytosolic DNA sensors and adapter proteins in type 1 interferon signaling. The terms “STING” and “interferon gene stimulator” refer to any form of the STING protein, as well as variants, isoforms, and species homologs that maintain at least some of the activity of STING. Unless otherwise indicated, such as by a specific reference to human STING, STING includes all mammalian species of the naturally occurring STING sequence, and for example, human, monkey, and mouse STING are also known as -TMEM173.

[0331] As used herein, “STING agonist” means any molecule that, upon binding 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, pharmaceuticals, and uses of the present invention include, for example, nucleic acid ligands that bind to STING.

[0332] Examples of STING agonists useful in the treatment methods, pharmaceuticals, and uses of the present invention include various immunostimulant nucleic acids such as synthetic double-stranded DNA, synthetic cyclic dinucleotides (CDNs) such as cyclic di-GMP, cyclic-GMP-AMP (cGAMP), MK-1454, and ADU-S100 (MIW815), and small molecules such as WO2019027858, WO20180093964, WO2017175156, and WO2017175147.

[0333] Therapeutic antibodies can exhibit specificity for a variety of different antigens. For example, a therapeutic antibody can target tumor-associated antigens, and its binding to the antigen promotes the death of cells expressing the antigen. In other cases, a therapeutic antibody can target antigens on immune cells, and its binding prevents the downregulation of the activity of cells expressing the antigen (and thereby promotes the activity of those cells). In some situations, therapeutic antibodies can function through multiple different mechanisms (e.g., 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 the antigen-expressing cells).

[0334] In another embodiment, such additional anticancer therapies include antibodies that would be blockading or inhibitory at the target: CTLA-4 (including ipilimumab or tremelimumab), PD-1 or PD-L1 (including atezolizumab, avelumab, semiprimab, durvalumab, nivolumab, sasamrimab, or pembrolizumab), LAG-3, TIM-3, or TIGIT.

[0335] In another embodiment, such additional anticancer agents include antibodies that are agonists of 4-1BB, OX40, GITR, ICOS, or CD40.

[0336] In another embodiment, the anti-cancer treatment may be CAR-T cell therapy.

[0337] 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, auristatin, drastatin, combretastatin, duocalmycin, pyrrolobenzodiazepine dimers, indolino-benzodiazepine dimers, engine, geldanamycin, meitansine, puromycin, taxanes, vinca alkaloids, camptothecin, tubulsin, hemiastalin, spriseostatin, prazienolides, and their stereoisomers, isoconforms, analogs, or derivatives. Examples of immunomodulatory agents that can be incorporated into ADCs include ganciclovir, etanercept, tacrolimus, sirolimus, voclosporine, cyclosporine, rapamycin, cyclophosphamide, azathioprine, mycophenolate mofetil, methotrexate, glucocorticoids and their analogues, cytokines, stem cell growth factors, lymphotoxins, tumor necrosis factor (TNF), hematopoietic factors, and interleukins (e.g., interleukin-1 (IL-1), IL-1). This includes 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, interferon-beta, and interferon-gamma), stem cell growth factors referred to as "S1 factors," erythropoietin and thrombopoietin, or combinations thereof.

[0338] Examples of additional therapeutic antibodies may include the following antigens, where exemplary antibodies targeting those antigens are also included below (in parentheses following the antigen). The following antigens may also be referred to herein as “target antigens.” Target antigens for therapeutic antibodies herein include, for example: 4-1BB (e.g., utomirumab); 5T4; A33; alpha-folate receptor 1 (e.g., milbetuximab / solabtansine); Alk-1; BCMA [see, for example, US9969809]; BTN1A1 (see, for example, WO2018222689); CA-125 (e.g., abagomomab); carboanhydrase IX; CCR2; CCR 4 (e.g., mogamulizumab); CCR5 (e.g., leronlimab); CCR8; CD3 [e.g., blinatumomab (CD3 / CD19 bispecificity), CD3 / P-cadherin bispecificity, CD3 / BCMA bispecificity]; CD19 (e.g., blinatumomab, MOR208); CD20 (e.g., ibritumomab / tiuxetan, obinutuzumab, ofatumumab, rituximab, ubrituximab); CD22 (inotuzumab... Ozogamicin, Moxetumomab Pasdotox); 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; C D63; CD79 (e.g., polatuzumab vedotin); CD80; CD123; CD276 / B7-H3 (e.g., omblutamab); CDH17; CEA; ClhCG; CTLA-4 (e.g., ipilimumab, tremelimumab), CXCR4; desmoglein 4; DLL3 (e.g., lovalpituzumab tesirin); DLL4; E-cadherin; EDA; EDB; EFNA4; EGFR (e.g., cetuximab, depatuxizumab mahodotin, nesitumumab, panitumumab); EGFRvIII; endothialin; EpCAM (e.g., oportuzumab monatox); FAP; fetal acetylcholine receptor;FLT3 (see, for example, WO2018 / 220584); GD2 (see, for example, dinutuximab, 3F8); GD3; GITR; GloboH; GM1; GM2; HER2 / neu [see, for example, margetuximab, pertuzumab, trastuzumab; ado-trastuzumab emtansine, trastuzumab duocalmazine, see, US8828401]; HER3; HER4; ICOS; IL-10; ITG-AvB6; LAG-3 (e.g., relatrimab); 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., olaratumab);Plasma cell antigen;PolySA;PSCA;PSMA;PTK7 [See US9409995];Ror1;SAS;SCRx6;SLAMF7 (e.g., elotuzumab);SHH;SIRPa (e.g., This includes 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); and Wue-1.

[0339] Exemplary imaging agents that may be included in ADCs include fluorescein, rhodamine, lanthanide phosphors, and their derivatives, or radioisotopes bound to chelating agents. Examples of fluorophores, but not limited to these, include fluorescein isothiocyanate (FITC) (e.g., 5-FITC), fluorescein amidite (FAM) (e.g., 5-FAM), eosin, carboxyfluorescein, erythrosine, 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 sulforodamine (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).

[0340] Exemplary therapeutic proteins that may be included in ADCs include toxins, hormones, enzymes, and growth factors.

[0341] Exemplary biocompatible polymers that may be incorporated into ADCs include water-soluble polymers such as polyethylene glycol (PEG) or its derivatives, and zwitterion-containing biocompatible polymers (e.g., phosphorylcholine-containing polymers).

[0342] Exemplary biocompatible polymers that may be incorporated into ADCs include antisense oligonucleotides.

[0343] The present invention also relates to the use of radiation in combination with any anticancer drug administered herein. More specifically, the compounds of the present invention can be administered in combination with additional treatments such as radiotherapy and / or chemotherapy.

[0344] These drugs and the compounds of the present invention can be combined with pharmaceutically acceptable vehicles such as physiological saline, Ringer's solution, dextrose solution, and the like. Specific dosing regimens, i.e., dosage, timing, and repetitions, will depend on the specific individual and their medical history.

[0345] kit Another aspect of the present invention provides a kit comprising the compound of the present invention or a pharmaceutical composition comprising the compound of the present invention. The kit may include, in addition to the compound of the present invention or its pharmaceutical composition, a diagnostic or therapeutic agent. The kit may include instructions for use in a diagnostic or therapeutic method. In some embodiments, the kit comprises the compound or its pharmaceutical composition and a diagnostic agent. In other embodiments, the kit comprises the compound or its pharmaceutical composition and one or more therapeutic agents.

[0346] In another embodiment, the present invention includes a kit suitable for use in carrying out the treatment methods described herein. In one embodiment, the kit contains a first dosage form comprising one or more compounds of the present invention in sufficient quantities to carry out the method of the present invention. In another embodiment, the kit comprises one or more compounds of the present invention in sufficient quantities to carry out the method of the present invention, and a container for dosage and a container for dosage.

[0347] Synthesis method The compounds of the present invention can be synthesized by synthetic routes involving processes similar to those well known in the chemical field, particularly in light of the description contained herein. 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 from which one or more scientific importance or commercial needs arise. Thus, such compounds may be one or more of the following: 1) commercially available; 2) reported in the literature; or 3) prepared by those skilled in the art from other commercially available substances using materials reported in the literature.

[0348] For illustrative purposes, the reaction scheme shown below provides a promising route for synthesizing the compounds of the present invention, as well as 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 see that the compounds of the present invention can be synthesized using other synthetic routes. While specific starting materials and reagents are discussed below, they can be substituted with other starting materials and reagents to obtain one or more derivatives or reaction conditions. In addition, many of the compounds prepared by the methods described below can be further modified using conventional chemistry well known to those skilled in the art in light of this disclosure.

[0349] Those skilled in the art will see that the experimental conditions described in the following scheme are examples of suitable conditions for performing the transformations shown, and that it may be necessary or desirable to change the exact conditions used to prepare the compounds of the present invention. Furthermore, they will see that it may be necessary or desirable to perform the transformations in a different order than those described in the scheme, or to change one or more of the transformations, in order to obtain the desired compounds of the present invention.

[0350] In preparing the compounds of the present invention, it should be noted that some of the preparation methods useful for preparing the compounds described herein may require the protection of distant 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 properties of the distant functional groups and the conditions of the preparation method. Those skilled in the art will readily determine the need for such protection. The use of such protection / deprotection methods is also within the scope of skill in the art. For an overview of protecting groups and their uses, see March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 8th edition.

[0351] 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 parts of the molecule. Therefore, such functional groups can be protected with a suitable protecting group (PG) that can be removed in a later step. Suitable protecting groups for amine and carboxylic acid protection include protecting groups commonly used in peptide synthesis (such as N-tert-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz), and 9-fluorenylmethyleneoxycarbonyl (Fmoc) for amines, and lower alkyl or benzyl esters for carboxylic acids), which are generally not chemically reactive under the reaction conditions described and can typically be removed without chemically altering other functional groups in the compounds of the present invention.

[0352] General experimental details 1 H and 19 The 1F nuclear magnetic resonance (NMR) spectra were recorded using a Bruker XWIN-NMR (400 or 700 MHz) spectrometer. 1 H and 19 F resonance has been reported in parts per million (ppm) from tetramethylsilane to low magnetic fields. 1¹H NMR data are reported as multiplexing (e.g., s, singlet; d, doublet; t, triplet; q, quadruplet; quint, quintlet; dd, doubledublet; dt, tripledublet; br s, broad singlet). For spectra obtained with CDCl3, DMSO-d6, and CD3OD, residual protons (7.27, 2.50, and 3.31 ppm, respectively) were used as an internal reference. All observed coupling constants, J, are reported in Hertz (Hz). Exchangeable protons are not always observed.

[0353] Optical activity was determined using a Jasco P-2000 or Rudolph Autopol IV polarimeter. All final compounds were purified to a purity of ≥95% unless otherwise specified. Where absolute stereochemistry was unknown, the software-generated names were modified to include the prefixes (+)- and (-)- according to the optical activity, and (R) to indicate relative stereochemistry. * / S * ) Signs are used.

[0354] Mass spectra, MS(m / z), were recorded using either electrospray ionization (ESI) or atmospheric pressure chemical ionization (APCI). Unless otherwise stated, the presented m / z data are isotope-based. 19 F, 35 Cl, 79 Br and 127 This concerns I.

[0355] The nomenclature is generated and written within Perkin Elmers Chemdraw 18.0.0.231, as specified by IUPAC (International Union for Pure and Applied Chemistry). The nomenclature rules provided in Perkin Elmers Chemdraw 18.0.0.231 are well known to those skilled in the art, and are considered to be in general conformity with the IUPAC (International Union for Pure and Applied Chemistry) recommendations and CAS Index rules regarding organic chemical nomenclature.

[0356] Abbreviation: CH3CN is acetonitrile, AQ is water-based; Bn is benzyl; Boc is a tert-butoxycarbonyl; Boc2O is ditert-butyl dicarbonate; br is broad; tBu is tert-butyl; °C is the unit of temperature in Celsius; CDCl3 is deuterated chloroform; δ is a chemical shift; d is a double line; dd is a double double line; ddd is a double dt is a double line with a triple line; DCM is dichloromethane; it is methylene chloride; DHP is dihydropyran, DIPEA is an 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; EA is ethyl acetate, ee is the enantiomer excess; ESI is electrospray ionization; Et2O is diethyl ether;  is ethyl acetate; EtOH is ethanol; Et3N is triethylamine; g stands for grams; HPLC is high-pressure liquid chromatography; hr is time; L stands for liter; LCMS is a liquid chromatography-mass spectrometry method; m is a multiline; M is a mole; m-CPBA is 3-chloroperbenzoic acid; MeOD_d4 is deuterated methanol; MeOH is methanol; 2-MeTHF is 2-methyltetrahydrofuran; mg stands for milligram; MHz stands for megahertz; min is minutes; mL stands for milliliter; An mmol is a millimolecule; mol is a mole; MOM is a methoxymethyl ether group; MS(m / z) is the mass spectral peak; NMR is nuclear magnetic resonance; Pd / C is palladium on carbon; Pd(dppf)Cl2 is [1,1'-bis(diphenylphosphino)ferrocene]di It is chloropalladium(II); PE stands for petroleum ether; pH is the hydrogen ion concentration; ppm is parts per million; psi is pounds per square inch; q is a quadruple line; rpm stands for revolutions per minute; rt is room temperature; RT is the retention time; RuPhos Pd G3 is (2-dicyclohexylphosphin-2',6'-diisopropoxy-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) methanesulfonate (CAS Number: 1445085-77-7); s is a single line; SEMCl is 2-(trimethylsilyl)ethoxymethyl chloride; SEM is 2-(trimethylsilyl)ethoxymethyl; SFC is supercritical fluid chromatography; t is a triple line; TBAF is tert-butylammonium fluoride; TFA is trifluoroacetic acid; THF is tetrahydrofuran; THP is tetrahydropyran; TLC is thin-layer chromatography; TMSCN is a trimethylsilyl cyanide; TsOH is p-toluenesulfonic acid; TsCl is p-toluenesulfonyl chloride; μL is a microliter; μmol stands for micromoles.

[0357] The scheme described below 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 scheme, the general methods for preparing the compounds are shown in either racemic or enantioenriched forms. It will be apparent to those skilled in the art that all synthetic transformations can be carried out in exactly the same manner, whether the substance is enantioenriched or racemic. Furthermore, the resolution into the desired optically active substance can be carried out at any desired point in the sequence using well-known methods, such as those described herein and in the chemical literature.

[0358] General method:

[0359] [ka]

[0360] As illustrated in Scheme 1, 4,5,7-trichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidine can be treated with an amino alcohol (cyclic or acyclic) in a suitable solvent (such as THF) in the presence of an effective base (such as DIPEA) to obtain an adduct via a SnAr reaction at the 4-chloro position, followed by treatment with a strong base such as LiOtBu to form a tetracycle. Stannane formation can be achieved using dialkyltin reagents and palladium catalysis. Still reactions with suitable protected indazole bromides using a palladium catalyst (such as tetrakis) and copper iodine in a suitable solvent such as dioxane introduce an indazole group. Oxidation from the 2-thiomethyl group to a sulfone can be achieved using an oxidizing agent (such as Oxone) in a buffered aqueous solvent containing NaHCO3 and either acetone or methyl ethyl ketone. The resulting sulfone group can be replaced with an alcohol nucleophile (((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol, CAS# 2097518-76-6, etc.) in a suitable solvent (CH3CN or DCM, etc.) using a suitable base (LiHMDS or LiOTMS, etc.). Alternatively, a thioether or amino product can be obtained using a thiol or amine. Deprotection is achieved by standard methods known in the art (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. This scheme is representative and illustrates a general exemplary route.

[0361] [ka]

[0362] As illustrated in Scheme 2, an alternative method achieves coupling of indazole and the nucleus using a one-pot procedure that avoids the Stannane reaction. The chloronucleus and indazole are treated with a boron reagent (such as B2pin2) and a suitable base (such as CsF) in a solvent (such as DMF) at high temperature with a suitable catalyst (such as CataCXium A Pd G3) to obtain a directly coupled product. This can then be included in the examples according to Scheme 1.

[0363] [ka]

[0364] Alternatively, the Suzuki method is used to achieve coupling of indazole with the tetracyclic nucleus, as illustrated in Scheme 3. Conversion of aryl bromide to boronic acid ester is achieved under standard conditions known in the art, such as treatment with B2Pin2 in a suitable solvent (such as tert-amyl alcohol) together with a palladium catalyst (such as palladium acetate), a base (such as potassium phosphate), and an additional phosphine ligand (such as triphenylphosphine). Coupling of the boronic acid ester with the chlorotetracyclic ester is achieved using a palladium catalyst together with a phosphine ligand and a suitable base in a suitable solvent containing water. The resulting intermediate can then be incorporated into the corresponding examples as detailed in Scheme 1.

[0365] [ka]

[0366] As illustrated in Scheme 4, the indazole intermediate is treated with a chlorinated oxidizing agent (such as N-chlorosuccinimide, NCS) to obtain a 3-chloroindazole derivative as a mixture of an oxidized thioether and a 2-chloro nucleoproduct. This crude mixture can then be incorporated into the examples according to Scheme 1 by deprotection with SnAr as described.

[0367] [ka]

[0368] As illustrated in Scheme 5, a thioether was oxidized with either oxone or mCPBA to obtain a sulfone, which was then treated with a nucleophile (alcohol) and a suitable base (LiOTMS) in a solvent (such as acetonitrile) to obtain an adduct via a SnAr reaction. Subsequent coupling of indazoleboronic acid or boronate was achieved by Suzuki Chemical using palladium catalysis under conditions known in the art.

[0369] The variables and substituents in Scheme 1, Scheme 2, Scheme 3, Scheme 4, and Scheme 5 are the same as those defined in the embodiments of formulas (I), (II), and (III) herein.

[0370] 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-commercial intermediates is provided as a further embodiment of the present invention.

[0371] Synthesis of intermediates: Intermediate 1(I-1): 4,5,7-Trichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidine

[0372] [ka]

[0373] Step 1: Synthesis of 4-bromo-2,6-dichloro-5-fluoropyridine-3-carboxylic acid, I-1a. Diisopropylamine (44.1 mL, 314 mmol) was dissolved in THF (300 mL), and the solution was cooled to -78°C. n-BuLi (2.5 M in hexane, 114 mL, 286 mmol) was added over 15 minutes. The mixture was stirred for 45 minutes, and 2,6-dichloro-5-fluoropyridine-3-carboxylic acid (CAS: 82671-06-5, 30 g, 143 mmol) was added as a solution in THF (75 mL) over 6 minutes. The mixture was stirred at -78°C for 30 minutes. 1,2-dibromo-1,1,2,2-tetrachloroethane (CAS: 630-25-1, 69.8 g, 214 mmol) was added as a solution in THF (120 mL) over 10 minutes. The reaction mixture was maintained at -78°C for 2 hours, and the reaction mixture was checked by LC-MS. A new peak with MH=242 (product - CO2H) was observed (negative ion mode). The mixture was quenched by adding water (120 mL). After stirring at -78°C for 10 minutes, the cooling bath was removed and 6N HCl (90 mL) was added. The pH=1 aqueous layer was extracted with RINKAN (×3). The combined organic extract was washed with brine (×2) and dried over MgSO4. The solvent was removed to obtain a solid, which was stirred in heptane (250 mL) for 1 hour to remove the tetrachloroethylene byproduct. After filtration, the solid was washed with heptane (3 × 100 mL) and dried to obtain 30.3 g (73%) of 4-bromo-2,6-dichloro-5-fluoropyridine-3-carboxylic acid, I-1a, as a cream-colored solid. 19 F NMR (376 MHz, DMSO) d -114.17.

[0374] Step 2: Synthesis of methyl N-(4-bromo-2,6-dichloro-5-fluoropyridine-3-carbonyl)carbamimidothioate, I-1b. A solution of 4-bromo-2,6-dichloro-5-fluoropyridine-3-carboxylic acid, I-1a (30.2 g, 104 mmol) was suspended in DCM (420 mL). Oxalyl chloride (25.0 mL, 300 mmol), followed by DMF (40 mg), was added. After stirring for 2 hours, a solid was still present and bubbles could still be observed forming. The mixture was then stirred overnight (16 hours). After stirring at room temperature for 16 hours, the solid had dissolved and the mixture had become a yellow solution. The solvent was removed under vacuum to obtain 33.3 g of the acid chloride as a light brown solid. In another 500 mL round-bottom flask, methylimidothiocarbamate sulfate (33.2 g, 177 mmol) was stirred with semi-saturated Na2CO3 (80 mL) to obtain a clear solution. Et2O (60 mL) was added to this solution and it was cooled to 10°C. Next, the I-1a acid chloride was slowly added as a solution in Depositphotos (120 mL), and the temperature was monitored with an internal thermometer. A very slight exothermic reaction was observed, and after the addition was complete, the ice bath was removed. After warming to room temperature, the mixture was stirred for 30 minutes, during which time the consumption of the acid chloride was monitored using negative mode ionization, and we waited for any further I-1a to be gone (hydrolysis occurs during LCMS, yielding acid). After the reaction was complete, the formation of a clean product was observed, and a new peak was observed at M+H=360 with multiple halogen patterns. The mixture was partitioned between water (100 mL) and Depositphotos (150 mL), and the aqueous layer was extracted with Depositphotos (×2). The combined organic extracts were washed with saturated NaHCO3 (×1), dried on MgSO4, and concentrated to obtain 35 g (93%) of methyl N-(4-bromo-2,6-dichloro-5-fluoropyridine-3-carbonyl)carbamimidothioate, I-1b, as a light brown solid, which was used in the next reaction without further purification. 1 H NMR (400 MHz, DMSO-d6) δ 9.03 (br s, 1 H), 9.53 (br s, 1 H), 2.40 (s, 3 H).

[0375] Step 3: Synthesis of 5,7-dichloro-8-fluoro-2-(methylsulfanyl)pyrido[4,3-d]pyrimidine-4(3H)-one, I-1c. N-(4-bromo-2,6-dichloro-5-fluoropyridine-3-carbonyl)carbamimidothioate, I-1b (14.7 g, 40.7 mmol) was dissolved in DMF (45 mL), and DIEA (14.2 mL, 81.4 mmol) was added. The reaction mixture was heated to 95 °C for 3 hours under N2, at which point LC-MS analysis showed a clean transformation of the product. After cooling to room temperature, the mixture solution was poured into aqueous pH 5 buffer and 100 g of ice, and the resulting solution was adjusted to pH 3 using 6N HCl. After addition to the cold aqueous solution, a pale yellow solid precipitated from the solution. This precipitate was collected in a Buchner funnel, washed with water (×3), and dried to obtain 5,7-dichloro-8-fluoro-2-(methylsulfanyl)pyrido[4,3-d]pyrimidine-4(3H)-one, I-1c 9.6 g (84%). 19 F NMR (376 MHz, DMSO) d 135.4.

[0376] Step 4: Synthesis of 4,5,7-trichloro-8-fluoro-2-(methylsulfanyl)pyrido[4,3-d]pyrimidine, I-1. 5,7-Dichloro-8-fluoro-2-(methylsulfanyl)pyrido[4,3-d]pyrimidine-4(3H)-one, I-1c (5.6 g, 20 mmol) was added to a flask, and DIEA (7.1 mL, 28.6 mmol) was added, and the suspension was cooled to 0°C under N2. POCl3 (30 mL, 320 mmol) was added all at once, and the ice bath was removed. The mixture was then heated to 90°C for 4 hours. LCMS analysis (of the sample dissolved in MeOH) showed M+H=294 and two monomethanol adducts with a Cl2 isotope pattern. Excess POCl3 was removed under vacuum by displacing it with a mixture of toluene and DCM (×2). After removing all volatile substances, the resulting orange solid was dry-loaded onto an 80g ISCO silica column and purified using a gradient of 0–100% siRNA in heptane, maintaining 100% siRNA over 7 column volumes as the product slowly leached out of the column. The fraction was concentrated to obtain 4,5,7-trichloro-8-fluoro-2-(methylsulfanyl)pyrido[4,3-d]pyrimidine, I-1 (5.7g, 95%) as an orange solid. 13 C NMR (101 MHz, DMSO-d6) δ ppm 165.5, 157.2, 148.6, 146.7, 146.0, 143.2, 137.4, 137.3, 114.8, 12.9; 19 F NMR (376 MHz, DMSO) d -135.5.

[0377] Intermediate 2(I-2): (S)-2-chloro-1-fluoro-12-(methylthio)-5a,6,9,10-tetrahydro-5H,8H-4,7-dioxa-3,10a,11,13-tetraazanaphtho[1,8-ab]heptalene

[0378] [ka] 4,5,7-Trichloro-8-fluoro-2-(methylsulfanyl)pyrido[4,3-d]pyrimidine (I-1, 450 mg, 1.51 mmol) was suspended in CH3CN (10 mL). DIPEA (276 μL, 1.59 mmol) was added, and the suspension was cooled to 0°C under N2. In a separate vial, [(3R)-1,4-Oxazepan-3-yl]methanol hydrochloride (CAS1262409-55-1-HCl salt, 232 mg, 1.39 mmol) was suspended in DCM (1 mL), and DIPEA (276 μL, 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 a flask containing the cold solution of I-1. After approximately 45 minutes at 0°C, LCMS analysis indicated that the initial reaction was complete. LiOtBu (1M 4.5 mL, 4.5 mmol in THF) was added dropwise, and the ice bath was removed. The ice bath was replaced with an oil bath, and the reaction mixture was heated at 50°C for 30 minutes. LC-MS analysis indicated that the cyclization step was complete. The solution was cooled to room temperature and evaporated. A saturated aqueous solution of NaHCO3 (10 mL) was added, and the mixture was extracted with DCM (3 × 30 mL). The combined organic extract was dried over Na2SO4 and evaporated. The process described above was repeated on the same scale, with the same observations and results. The crudes from both reactions were combined and purified using flash chromatography with elution in a gradient of 0–100% siRNA in heptane, and DCM was used to load the crude onto a silica cartridge. The fraction containing the desired product was stored and concentrated to obtain (S)-2-chloro-1-fluoro-12-(methylthio)-5a,6,9,10-tetrahydro-5H,8H-4,7-dioxa-3,10a,11,13-tetraazanaphtho[1,8-ab]heptalene, I-2 483 mg (70%) as a light brown solid. 1 ¹H 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).

[0379] Intermediate 3(I-3): (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

[0380] [ka] 4,5,7-Trichloro-8-fluoro-2-(methylsulfanyl)pyrido[4,3-d]pyrimidine (I-1, 1.25 g, 3.70 mmol) was suspended in CH3CN (24 mL), and DIEA (0.668 mL, 3.83 mmol) was added. The mixture was cooled to 0°C, and (S)-piperidine-2-ylmethanol (421 mg, 3.65 mmol) was added as a solution in THF (18 mL). As observed by LC-MS, the first nitrogen-carbon bond was formed after 8 minutes. LiOtBu (877 mg, 11.0 mmol) was added as a solution in THF (22 mL), and the mixture was heated to 50°C. After 4 hours at 50°C, LC-MS analysis showed conversion to the product. The reaction mixture was then diluted with 200 mL of water, and the product was extracted with DCM (50 mL x 4). The combined organic extracts were dried over Na2SO4, filtered, and evaporated to obtain a crude solid. Purification was achieved by flash chromatography with elution under a gradient of 0-10% MeOH in DCM to obtain (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, I-3 (1.13 g, 91%). 1 H NMR (chloroform-d, 400 MHz) d 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] + .

[0381] Intermediate 4(I-4): (S)-4-fluoro-2-(methylthio)-5-(tributylstannyl)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene

[0382] [ka] To a solution of (S)-2-chloro-1-fluoro-12-(methylthio)-5a,6,9,10-tetrahydro-5H,8H-4,7-dioxa-3,10a,11,13-tetraazanaphtho[1,8-ab]heptalene (I-2, 7.0 g, 19.6 mmol) in toluene (980 mL), 1,1,1,2,2,2-hexabutyldistannanane (11.38 g, 9.9 mL, 19.6 mmol), CataCXium A (880 mg, 2.454 mmol), and Pd(OAc)2 (276 mg, 1.226 mmol) were added at 20 °C. The reaction mixture was stirred at 90 °C for 16 hours under an N2 atmosphere. LC-MS showed that the desired product had formed. The reaction mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography (Combi-Flash, 80g silica gel, 0%-30% phenylethylamine in petroleum ether) to obtain (S)-4-fluoro-2-(methylthio)-5-(tributylstannyl)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene, I-4 (5.65g, 47.1%) as a yellow oily substance. 1 H NMR (400 MHz, CDCl3) δ 5.20 (ddd, J = 13.7, 6.9, 2.6 Hz, 1H), 4.64 (dd, J = 13.4, 4.4 Hz, 1H), 4.37 (d, J = 13.3 Hz, 1H), 4.20 (dd, J = 12.6, 3.5 Hz, 1H), 4.06 - 3.97 (m, 2H), 3.74 (dd, J = 12.6, 9.9 Hz, 1H), 3.39 - 3.24 (m, 2H), 2.60 (s, 3H), 2.26 - 2.14 (m, 1H), 2.03 - 1.90 (m, 1H), 1.63 - 1.51 (m, 6H), 1.38 - 1.19 (m, 12H), 0.87 (t, J = 7.3Hz, 9H). 19 F NMR (376 MHz, CDCl3) δ -132.84. MS: 613.2 [M+H] + .

[0383] Intermediates 5i and 5ii (I-5i and I-5ii): 4-bromo-6-chloro-5-cyclopropyl-2-(tetrahydro-2H-pyran-2-yl)-2H-indazole and 4-bromo-6-chloro-5-cyclopropyl-1-(tetrahydro-2H-pyran-2-yl)-3a,7a-dihydro-1H-indazole

[0384] [ka]

[0385] Step 1: Synthesis of 4-chloro-5-cyclopropyl-2-fluoroaniline (I-5a) To a solution of 5-bromo-4-chloro-2-fluoroaniline (CAS111010-07-2, 8.06 g 35.9 mmol) in 1,4-dioxane (200 mL), cyclopropylboronic acid (4.63 g 53.9 mmol), K3PO4 (15.2 g 71.8 mmol), and PdCl2 (dppf) (3.15 g 4.31 mmol) were added. The mixture was stirred at 90°C under N2 for 8 hours. TLC analysis indicated that the reaction was complete. Water (100 mL) was added, and the product was extracted in ELISA (2 × 150 mL). The combined organic extract was washed with brine, and the organic solvent was concentrated. The crude product was purified by flash chromatography using a 0-40% ethyl acetate gradient in petroleum ether to obtain 4-chloro-5-cyclopropyl-2-fluoroaniline, I-5a (5.4 g, 81%) as a yellow oil. 1 H NMR (400MHz, CDCl3) δ 7.02 (d, J=10.5 Hz, 1H), 6.38 (d, J=9.3 Hz, 1H), 4.24 - 2.95 (m, 2H), 2.13 - 2.01 (m, 1H), 1.01 - 0.89 (m, 2H), 0.64 - 0.48 (m, 2H).

[0386] Step 2: Synthesis of 3-bromo-1-chloro-2-cyclopropyl-5-fluoro-4-iodobenzene (I-5b) A solution of 4-chloro-5-cyclopropyl-2-fluoroaniline (I-5a, 5.40 g 29.1 mmol) in DMF (50 mL) was cooled to 0°C, and NBS (5.18 g 29.1 mmol) was gradually added under N2. The mixture was warmed to room temperature and stirred for 1 hour. TLC analysis showed the consumption of the starting material. The mixture was diluted with saturated NaHCO3 aqueous solution, and the product was extracted with HCl (2 × 150 mL). The combined organic extract was washed with brine, dried, and filtered to obtain a crude bromoaniline intermediate, which was purified by flash chromatography with elution under a 0-40% HCl gradient in petroleum ether to obtain the bromoaniline intermediate (7.2 g, 93.6%) as a brown oily substance. Bromoaniline (6.55 g, 24.8 mmol) was added to a solution of concentrated H2SO4 (17 mL) in water (65 mL), and the solution was cooled to 5°C. NaNO2 (1.88 g, 27.2 mmol) was added dropwise to water (6 mL). The resulting mixture was stirred at 5°C for 20 minutes. The resulting diazo solution was added at 5°C to a solution of KI (16.4 g, 99.0 mmol) in water (14 mL). The reaction mixture was stirred at 5°C for 20 minutes, and then warmed to room temperature. Stirring was continued at room temperature for 18 hours. TLC analysis showed that bromoaniline had been almost completely consumed. The mixture was quenched with water (100 mL), and the product was extracted with ELISA (2 × 110 mL). The combined organic extracts were washed with saturated aqueous Na2SO3 and brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. Purification by flash chromatography yielded 3-bromo-1-chloro-2-cyclopropyl-5-fluoro-4-iodobenzene, I-5b (6.03 g, 65%) as an oil. 1¹H NMR (400MHz, chloroform-d) d 7.14 (d, J=7.5 Hz, 1H), 1.86 - 1.74 (m, 1H), 1.30 - 1.21 (m, 2H), 0.78 - 0.72 (m, 2H).

[0387] Step 3: Synthesis of 2-bromo-4-chloro-3-cyclopropyl-6-fluorobenzaldehyde (I-5c) To a solution of 3-bromo-1-chloro-2-cyclopropyl-5-fluoro-4-iodobenzene (I-5b, 5.0 g 13 mmol) in THF (25 mL), n-BuLi (2.5 M, 5.3 mL, 13.3 mmol) was added dropwise under argon at -78°C. The mixture was stirred for 30 minutes, and anhydrous DMF (1.07 g 14.7 mmol) was added, while stirring was continued at -78°C for 20 minutes. TLC analysis showed consumption of the starting material and the formation of new spots. The mixture was quenched with 1N HCl (2 mL) and slowly warmed to room temperature. The reaction mixture was diluted with water (20 mL), and the product was extracted with RINKAN (2 × 150 mL). The combined organic extract was washed with water, dried, filtered, and concentrated. Purification by flash chromatography using a gradient of 0-40% siRNA in petroleum ether yielded 2-bromo-4-chloro-3-cyclopropyl-6-fluorobenzaldehyde, I-5c (2.55 g, 69%) as a yellow solid, and in addition, 3-bromo-1-chloro-2-cyclopropyl-5-fluoro-4-iodobenzene (I-5b, 1.03 g, 21%) was recovered. 1 H NMR (400MHz, DMSO-d6) δ 10.19 (s, 1H), 7.71 (d, J=10.3 Hz, 1H), 1.88 - 1.76 (m, 1H), 1.28 - 1.19 (m, 2H), 0.74 - 0.65 (m, 2H); LCMS (ESI) m / z: 277.0 [M+H]+.

[0388] Step 4: Synthesis of 4-bromo-6-chloro-5-cyclopropyl-2H-indazole (I-5d) A mixture of 2-bromo-4-chloro-3-cyclopropyl-6-fluorobenzaldehyde (I-5c, 2.05 g, 7.38 mmol) in 1,4-dioxane (25 mL) was mixed with 85% N2H4-H2O (2.21 g, 37.6 mmol) at 10 °C. The reaction mixture was heated to 90 °C for 16 hours. The mixture was combined with the crude product of the same reaction carried out separately. The combined reaction mixture was slowly added to water (50 mL) to obtain a creamy suspension, which was filtered and washed with water. The filtrate was dissolved in MTBE (150 mL), and the organic layer was dried over Na2SO4. After concentration, the substance was purified by flash chromatography eluting under a gradient of 0-30% siRNA in petroleum ether to obtain 4-bromo-6-chloro-5-cyclopropyl-2H-indazole, I-5d (1.88 g, 83%) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ 13.43 (br s, 1H), 8.02 (s, 1H), 7.69 (s, 1H), 1.84 (tt, J=5.6, 8.3 Hz, 1H), 1.22 - 1.17 (m, 2H), 0.73 - 0.68 (m, 2H); LCMS (ESI) m / z: 271.9 [M+H]+.

[0389] Step 5: Synthesis of 4-bromo-6-chloro-5-cyclopropyl-2-(oxan-2-yl)-2H-indazole (I-5i) and 4-bromo-6-chloro-5-cyclopropyl-2-(oxan-2-yl)-2H-indazole (I-5ii) To a solution of 4-bromo-6-chloro-5-cyclopropyl-2H-indazole I-5d (2.1 g, 7.9 mmol) in DCM (40 mL), dihydropyran (999 mg, 11.9 mmol) and p-TsOH (136 mg, 0.792 mmol) were added. The reaction mixture was stirred at 30°C for 16 hours. LC-MS analysis showed the formation of two products with the same mass. The reaction mixture was diluted with DCM (40 mL) and washed with saturated NaHCO3 aqueous solution (50 mL) and brine (50 mL). The organic extract was dried and concentrated to obtain a crude mixture, which was purified by flash chromatography with elution in a 0-10% siRNA gradient in petroleum ether to obtain 4-bromo-6-chloro-5-cyclopropyl-2-(oxan-2-yl)-2H-indazole (I-5i, 1.9 g, 68%) as a yellow rubbery substance, and 4-bromo-6-chloro-5-cyclopropyl-2-(oxan-2-yl)-2H-indazole (I-5ii, 335 mg, 12%) as a yellow rubbery substance. 1 ¹H NMR I-5i (400MHz, chloroform-d) d 8.21 - 8.12 (m, 1H), 7.73 (s, 1H), 5.65 (dd, J=3.0, 9.0 Hz, 1H), 4.20 - 4.12 (m, 1H), 3.80 (dt, J=3.1, 11.0 Hz, 1H), 2.29 - 2.12 (m, 2H), 2.09 - 2.01 (m, 1H), 1.88 - 1.82 (m, 1H), 1.80 - 1.69 (m, 3H), 1.25 - 1.19 (m, 2H), 0.85 - 0.79 (m, 2H), MS: 313.9 [M+H-cyclopropyl] + . 1 1H NMR I-5ii (400 MHz, CDCl3) δ 7.98 (s, 1H), 7.62 (s, 1H), 5.62 (dd, J = 9.1, 2.7 Hz, 1H), 4.06 - 3.95 (m, 1H), 3.80 - 3.67 (m, 1H), 2.54 - 2.41 (m, 1H), 2.19 - 2.01 (m, 2H), 1.89 - 1.65 (m, 5H), 1.24 - 1.17 (m, 2H), 0.81 - 0.72 (m, 2H). MS: 354.9, 356.9 [M+H] + .

[0390] Intermediate 6(I6):4-bromo-6-chloro-5-isopropyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole

[0391] [ka]

[0392] Step 1: Synthesis of 3-bromo-5-chloro-4-iodo-2-methylaniline (I-6b) To a stirred solution of 3-bromo-5-chloro-2-methylaniline (10 g, 45 mmol) in CH3CN (113 mL), indium(III) trifluoromethanesulfonate (12.7 g, 22.7 mmol), followed by ICl (7.36 g, 45.4 mmol), was added dropwise at 0°C. The reaction mixture was stirred at 0°C for 2 hours. Then, the mixture was heated to 25°C and stirred at 25°C for 16 hours. LC-MS showed the detection of the desired product. The reaction mixture was quenched with TEA (9.48 mL, 68 mmol) and then stirred for 15 minutes. The solid was removed by filtration on Celite. The filtrate cake was rinsed with siRNA (100 mL). The filtrate was collected, washed with saturated NaCl (70 mL), dried on Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by flash column chromatography (Combi-Flash, 120 g silica gel, 0-5% siRNA in petroleum ether) to obtain the title intermediate (9.5 g, 60%) as a solid. 1 H NMR (400 MHz, CDCl3) δ 6.85 (s, 1H), 3.82 (s, 2H), 2.38 (s, 3H). MS: 345.9, 347.8 [M+H]+ .

[0393] Step 2: Synthesis of 3-bromo-5-chloro-2-methyl-4-(propa-1-en-2-yl)aniline (I-6c) Pd(dppf)Cl2 (296 mg, 0.404 mmol) was added to a mixture of 3-bromo-5-chloro-4-iodo-2-methylaniline (1.4 g, 4.0 mmol), 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (1.02 g, 6.06 mmol), and K3PO4 (2.57 g, 12.1 mmol) in 1,4-dioxane (20 mL) and H2O (4 mL). The mixture was stirred at 80°C under nitrogen for 3 hours. LCMS showed that the starting materials were consumed and the desired compound was found. The mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography (Combi-Flash, 10 g silica gel, 0-7% siRNA in petroleum ether) to obtain the title intermediate (820 mg, 78%) as a yellow solid. 1 H NMR (400 MHz, DMSO) δ 6.73 (s, 1H), 5.41 (s, 2H), 5.31 - 5.23 (m, 1H), 4.80 - 4.68 (m, 1H), 2.15 (s, 3H), 1.90 (s, 3H). MS: 260.0, 262.0 [M+H] + .

[0394] Step 3: Synthesis of 3-bromo-5-chloro-4-isopropyl-2-methylaniline (I-6d) To a solution of 3-bromo-5-chloro-2-methyl-4-(prop-1-en-2-yl)aniline (500 mg, 1.92 mmol) in siRNA (14 mL) and isopropyl alcohol (14 mL), PtO2 (218 mg, 0.959 mmol) was added at 25°C. The reaction mixture was stirred at 45°C under a hydrogen atmosphere at 4 atm for 24 hours. LC-MS showed that 40% of the desired compound was found. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (Combi-Flash, 10 g silica gel, 0-12% siRNA in petroleum ether) to obtain the title intermediate (300 mg, 60% crude) as a brown oily substance. 1 H NMR (400 MHz, CDCl3) δ 6.67 (s, 1H), 3.95 - 3.82 (m, 3H), 2.28 (s, 3H), 1.37 (d, J = 7.1 Hz, 6H). MS: 262.0, 264.0 [M+H] + .

[0395] Step 4: Synthesis of 3-bromo-5-chloro-4-isopropyl-2-methylbenzenediazonium tetrafluoroborate (I-6e) BF3·Et2O (243 mg, 1.71 mmol, 0.217 mL) was dissolved in DCM (8 mL) and cooled to -5°C under a nitrogen atmosphere. A solution of 3-bromo-5-chloro-4-isopropyl-2-methylaniline (300 mg, 1.14 mmol) in DCM (1 mL) was added to the above reaction mixture and stirred at -5°C for 0.5 hours. Tert-butyl nitrite (177 mg, 1.71 mmol) in DCM (1 mL) was added dropwise. The reaction mixture was slowly heated to 25°C and stirred at the same temperature for 16 hours. LC-MS showed that the starting material was consumed and the desired product was detected. The reaction mixture was concentrated to 0.5 mL, and then MTBE (10 mL) was added. The suspension was filtered to obtain the title intermediate (290 mg, 70% crude) as a white solid. MS: 273.0, 275.0 [M] + .

[0396] Step 5: Synthesis of 4-bromo-6-chloro-5-isopropyl-1H-indazole (I-6f) To a mixture of 18-crown-6 (63.6 mg, 0.241 mmol) in chloroform (5 mL), KOAc (110 mg, 1.12 mmol) was added at 25°C. Then, 3-bromo-5-chloro-4-isopropyl-2-methylbenzenediazonium tetrafluoroborate (290 mg, 0.803 mmol) was slowly added. The reaction mixture was then stirred at 25°C for 1 hour. LC-MS detected the desired product and indicated that the starting material had been consumed. The reaction mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography (Combi-Flash, 4 g silica gel, 0-70% ethyl ether in petroleum ether) to obtain the title intermediate (120 mg, 55%) as a yellow, rubbery substance. 1 H NMR (400 MHz, CDCl3) δ 8.05 (s, 1H), 7.51 (s, 1H), 4.07 - 3.96 (m, 1H), 1.47 (d, J = 6.6 Hz, 6H). MS:273.0, 274.9 [M+H] + .

[0397] Step 6: Synthesis of 4-bromo-6-chloro-5-isopropyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole (I-6) To a solution of 4-bromo-6-chloro-5-isopropyl-1H-indazole (120 mg, 0.439 mmol) in DCM (7 mL), DHP (55.3 mg, 0.658 mmol) was added, followed by p-toluenesulfonic acid monohydrate (8.34 mg, 0.0439 mmol). The reaction mixture was stirred at 25°C for 12 hours. LC-MS showed that the starting material was consumed and the desired product was detected. The reaction mixture was diluted with DCM (20 mL) and washed with saturated NaHCO3 (30 mL) and brine (30 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (Combi-Flash, 4 g silica gel, 0-4% Â in petroleum ether) to obtain the title intermediate (130 mg, 83%) as a yellow solid. 1 H NMR (400 MHz, CDCl3) δ 7.96 (s, 1H), 7.60 (s, 1H), 5.62 (dd, J = 9.1, 2.5 Hz, 1H), 4.08 - 3.92 (m, 2H), 3.80 - 3.69 (m, 1H), 2.54 - 2.44 (m, 1H), 2.18 - 2.01 (m, 3H), 1.77 - 1.71 (m, 2H), 1.46 (d, J = 6.9 Hz, 6H).MS: 357.0, 359.0 [M+H] + .

[0398] Intermediate 7(I-7):(2S,7aR)-7a-(hydroxymethyl)-6-methylenehexahydro-1H-pyrrolidine-2-ol

[0399] [ka]

[0400] Step 1: Synthesis of 1-(tert-butyl)2-methyl(2R,4R)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-1,2-dicarboxylate (I-7b) To a solution of 1-(tert-butyl)2-methyl(2R,4R)-4-hydroxypyrrolidine-1,2-dicarboxylate (I-7a, 22 g, 90 mmol) in DMF (90 mL), imidazole (9.16 g, 135 mmol) and TBSCl (17.6 g, 117 mmol) were added at 0°C. The reaction mixture was stirred at 20°C for 16 hours. LC-MS showed that the starting material was consumed and the desired product was detected. The reaction mixture was concentrated under vacuum. The residue was diluted with water (200 mL), extracted with ELISA (100 mL), and washed with water (20 mL x 3). The combined organic layers were concentrated under reduced pressure. The residue was purified by flash column chromatography (Biotage, 120 g silica gel, 0-8% ethyl ether) to obtain 1-(tert-butyl)2-methyl(2R,4R)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-1,2-dicarboxylate (32 g, 99%) as a colorless oil. 1 H NMR (400 MHz, CDCl3) δ 4.43 - 4.25 (m, 2H), 3.70 (s, 3H), 3.60 (ddd, J = 27.1, 11.1, 5.4 Hz, 1H), 3.30 (ddd, J = 21.3, 11.1, 3.4 Hz, 1H), 2.36 - 2.20 (m, 1H), 2.14 - 2.04 (m, 1H), 1.47 and 1.41 (2s, 9H, subrotational and major rotational isomers, respectively), 0.85 and 0.84 (2s, 9H, subrotational and major rotational isomers, respectively), 0.06 - 0.01 (m, 6H). MS: 382.1 [M+Na] + .

[0401] Step 2: Synthesis of 1-(tert-butyl)2-methyl(2R,4R)-4-((tert-butyldimethylsilyl)oxy)-2-(2-(chloromethyl)allyl)pyrrolidine-1,2-dicarboxylate(I-7c) To a solution of 1-(tert-butyl)2-methyl(2R,4R)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-1,2-dicarboxylate (I-7b, 5.00 g, 13.9 mmol) in THF (15 mL), LiHMDS (16.7 mL, 1 M in hexane, 16.7 mmol) was added dropwise at -78°C. The mixture was stirred at -78°C for 30 minutes, and then 3-chloro-2-(chloromethyl)propa-1-ene (4.35 g, 34.8 mmol) in THF (17.3 mL) was added. The reaction mixture was slowly warmed to 20°C and stirred at 20°C for 16 hours. LC-MS showed that the starting material was consumed and the desired product was detected. The reaction mixture was quenched with saturated NH4Cl (aqueous solution) (10 mL) at 0°C. The resulting mixture was extracted with SiO2 (30 mL x 3), the combined organic layers were washed with water and brine, and dried over Na2SO4. After filtration, the filtrate was concentrated under vacuum. The residue was purified by flash column chromatography (Biotage, 80 g silica gel, 0-10% SiO2 in petroleum ether) to obtain 1-(tert-butyl)2-methyl(2R,4R)-4-((tert-butyldimethylsilyl)oxy)-2-(2-(chloromethyl)allyl)pyrrolidine-1,2-dicarboxylate (I-7c, 5.0 g, 80%) as a colorless oil. 1 ¹H NMR (400 MHz, CDCl3) δ 5.40 (d, J = 1.0 Hz, 1H), 5.11 and 5.10 (2 s, 1H, (principal and secondary isomers), 4.29 - 4.20 (m, 1H), 4.05 - 3.93 (m, 2H), 3.75 - 3.69 (m, 3H), 3.67 - 3.54 (m, 1H), 3.35 - 3.13 (m, 2H), 2.65 (dd, J = 14.4, 3.0 Hz, 1H), 2.34 - 2.16 (m, 1H), 2.14 - 2.01 (m, 1H), 1.45 and 1.44 (2 s, 9H, sub-rotational isomer and major rotational isomer, respectively), 0.86 and 0.85 (2 s, 9H, sub-rotational isomer and major rotational isomer, respectively), 0.04 - 0.00 (m, 6H). MS: 470.1 [M+Na] + .

[0402] Step 3: Synthesis of 1-(tert-butyl)2-methyl(2R,4R)-2-(2-(chloromethyl)allyl)-4-hydroxypyrrolidine-1,2-dicarboxylate (I-7d) To a solution of 1-(tert-butyl)2-methyl(2R,4R)-4-((tert-butyldimethylsilyl)oxy)-2-(2-(chloromethyl)allyl)pyrrolidine-1,2-dicarboxylate (I-7c, 5.0 g, 11 mmol) in THF (37.2 mL), TBAF (13.4 mL, 1 M in THF, 13.4 mmol) was added dropwise at 20°C. The reaction mixture was stirred at 25°C for 20 hours. LC-MS showed that the starting material was consumed and the desired product was detected. The reaction mixture was concentrated. The residue was dissolved in ELISA (100 mL) and washed with saturated NaHCO3 (aqueous solution) (30 mL x 7). The organic layer was dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by flash column chromatography (Biotage, 80 g silica gel, 45-55% ethyl ether in petroleum ether) to obtain 1-(tert-butyl)2-methyl(2R,4R)-2-(2-(chloromethyl)allyl)-4-hydroxypyrrolidine-1,2-dicarboxylate (I-7d, 3.0 g, 80%) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 5.42 (s, 1H), 5.06 (s, 1H), 4.21 (s, 1H), 4.04 - 3.92 (m, 2H), 3.90 - 3.73 (m, 4H), 3.43 - 3.24 (m, 2H), 2.58 (t, J = 13.6 Hz, 1H), 2.44 (ddd, J = 29.5, 14.5, 5.2 Hz, 1H), 2.11 (d, J = 15.2 Hz, 1H), 1.47 (s, 9H). MS: 356.5 [M+Na] + .

[0403] Step 4: Synthesis of 1-(tert-butyl)2-methyl(2R,4S)-2-(2-(chloromethyl)allyl)-4-((4-nitrobenzoyl)oxy)pyrrolidine-1,2-dicarboxylate(I-7e) DEAD (4.13 g, 23.7 mmol) was added to a solution of 1-(tert-butyl)2-methyl(2R,4R)-2-(2-(chloromethyl)allyl)-4-hydroxypyrrolidine-1,2-dicarboxylate (I-7d, 6.6 g, 20 mmol), 4-nitrobenzoic acid (3.97 g, 23.7 mmol), and PPh3 (6.22 g, 23.7 mmol) in THF (66 mL) under nitrogen at 0°C. The mixture was stirred at 20°C for 3 hours. LC-MS showed that the starting materials were consumed and the desired product was detected. The mixture was quenched with saturated NaHCO3 (water) solution (15 mL) and extracted with SiO2 (50 mL x 3). The combined organic layers were washed with water (100 mL) and brine (100 mL), dried over Na2SO4, filtered, and the filtrate was concentrated. The residue was purified by flash column chromatography (Biotage, 120 g silica gel, 15-25% ethyl ether in petroleum ether) to obtain methyl(2R,4S)-2-(2-(chloromethyl)allyl)-4-((4-nitrobenzoyl)oxy)pyrrolidine-2-carboxylate (I-7e, 9.0 g, 94%) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 8.31 (d, J = 8.8 Hz, 2H), 8.23 ​​- 8.14 (m, 2H), 5.55 - 5.41 (m, 1H), 5.32 and 5.24 (2 s, 1H, sub-rotational isomer and major rotational isomer, respectively), 5.07 and 5.04 (2 s, 1H, sub-rotational isomer and major rotational isomer, respectively), 4.16 - 4.05 (m, 3H), 3.78 and 3.76 (2 s, 3H, (main and sub-rotational isomers, respectively), 3.65 - 3.51 (m, 1H), 3.44 - 3.21 (m, 1H), 2.81 (dd, J = 22.0, 14.7 Hz, 1H), 2.64 - 2.43 (m, 2H), 1.47 (s, 9H). MS: 505.1 [M+Na] + .

[0404] Step 5: Synthesis of methyl(2R,4S)-2-(2-(chloromethyl)allyl)-4-((4-nitrobenzoyl)oxy)pyrrolidine-2-carboxylate(I-7f) To a solution of methyl(2R,4S)-2-(2-(chloromethyl)allyl)-4-((4-nitrobenzoyl)oxy)pyrrolidine-2-carboxylate (I-7e, 6.0 g, 12 mmol) in DCM (41 mL), TFA (9.5 mL) was added at 0°C. The mixture was stirred at 20°C for 16 hours. LC-MS showed that the starting material was consumed and the desired product was detected. The reaction mixture was then concentrated to obtain the crude product methyl(2R,4S)-2-(2-(chloromethyl)allyl)-4-((4-nitrobenzoyl)oxy)pyrrolidine-2-carboxylate (I7f, 8.2 g, crude) as a yellow oil. MS: 383.1 [M+H] + .

[0405] Step 6: Synthesis of methyl(6S,7aR)-2-methylene-6-((4-nitrobenzoyl)oxy)tetrahydro-1H-pyrrolidine-7a(5H)-carboxylate(I-7g) To a solution of methyl(2R,4S)-2-(2-(chloromethyl)allyl)-4-((4-nitrobenzoyl)oxy)pyrrolidine-2-carboxylate (I-7f, 200 mg, 0.522 mmol) in CH3CN (5 mL), K2CO3 (361 mg, 2.61 mmol) and KI (8.67 mg, 0.0522 mmol) were added at 25°C. The reaction mixture was stirred at 25°C for 1 hour. LC-MS showed that the starting material was consumed and the desired product was detected. The reaction mixture was filtered, and the filtrate was concentrated to obtain the product methyl(6S,7aR)-2-methylene-6-((4-nitrobenzoyl)oxy)tetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (I-7 g, 170 mg, 94%) as a pale yellow oil. 1 H NMR (400 MHz, CDCl3) δ 8.30 (d, J = 8.6 Hz, 2H), 8.16 (d, J = 8.6 Hz, 2H), 5.58 (s, 1H), 5.02 (s, 1H), 4.98 (s, 1H), 3.94 (d, J = 14.4 Hz, 1H), 3.77 (s, 3H), 3.62 (dd, J = 12.6, 5.1 Hz, 1H), 3.52 (d, J = 14.3 Hz, 1H), 3.15 (d, J = 16.2 Hz, 2H), 3.08 (dd, J = 12.6, 1.8 Hz, 1H), 2.87 (dd, J = 14.6, 6.7 Hz, 1H), 2.75 (d, J = 16.2 Hz, 1H), 2.20 (dd, J = 14.6, 2.7 Hz, 1H). MS: 347.1 [M+H] + .

[0406] Step 7: Synthesis of (2S,7aR)-2-hydroxy-6-methylenetetrahydro-1H-pyrrolidine-7a(5H)-carboxylic acid (I-7h) A solution of methyl (6S,7aR)-2-methylene-6-((4-nitrobenzoyl)oxy)tetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (I-7g, 500mg, 1.44 mmol) and K2CO3 (2000mg, 14.4 mmol) in methanol (10 mL) was stirred at 20°C for 2 hours. LC-MS showed that the starting material was consumed and the desired product was detected. The reaction mixture was filtered, and the filtrate was concentrated to obtain the crude product (2S,7aR)-2-hydroxy-6-methylenetetrahydro-1H-pyrrolidine-7a(5H)-carboxylic acid (I-7h, 300 mg, crude) as a pale yellow oil. MS: 184.1 [M+H] + .

[0407] Step 8: Synthesis of (2S,7aR)-7a-(hydroxymethyl)-6-methylenehexahydro-1H-pyrrolidine-2-ol (I-7) To a stirred solution of (2S,7aR)-2-hydroxy-6-methylenetetrahydro-1H-pyrrolidine-7a(5H)-carboxylic acid (I-7h, 250 mg, 1.36 mmol) in THF (6.8 mL), LiAlH4 (8.19 mL, 8.19 mmol, 1 M in THF) was added dropwise at 0°C. The resulting mixture was stirred at 60°C for 3 hours. LC-MS showed that the starting material was consumed and the desired product was formed. The reaction mixture was quenched with Na2SO4·10H2O solid (0.5 g) and stirred at 20°C for 2 hours. The suspension was filtered through Celite, and the filtrate was concentrated under vacuum. The residue was purified by flash column chromatography (Biotage, 4 g, 15-30% MeOH in DCM (1% NH4OH in MeOH)) to obtain (2S,7aR)-7a-(hydroxymethyl)-6-methylenehexahydro-1H-pyrrolidine-2-ol (I-7, 70 mg, 30%) as a pale yellow oil. 1 H NMR (400 MHz, CDCl3) δ 4.98 - 4.90 (m, 2H), 4.49 (dq, J = 10.5, 5.2 Hz, 1H), 3.68 (d, J = 15.4 Hz, 1H), 3.46 (dd, J = 14.6, 1.3 Hz, 1H), 3.31 (dd, J = 10.8, 5.3 Hz, 1H), 3.25 (s, 2H), 2.73 (dd, J = 10.8, 5.2 Hz, 1H), 2.63 (dd, J = 15.8, 1.4 Hz, 1H), 2.41 (dd, J = 15.8, 0.9 Hz, 1H), 2.29 (dd, J = 13.4, 6.7 Hz, 1H), 1.77 (dd, J = 13.4, 4.9 Hz, 1H). MS: 170.2 [M+H] + .

[0408] Intermediate 8(I-8): [(2Z,7aS)-2-(2-methoxyethylidene)tetrahydro-1H-pyrrolidine-7a(5H)-yl]methanol

[0409] [ka]

[0410] Step 1: Synthesis of ethyl(S)-2-(2-(tert-butoxy)-2-oxoethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate(I-8a) To a stirred solution of tert-butyl 2-(dimethoxyphosphoryl)acetate (2340 mg, 10.4 mmol) in THF (40 mL), NaH (417 mg, 10.4 mmol) was slowly added at 0°C. The suspension was stirred at 0°C for 10 minutes. Then, ethyl(S)-2,5-dioxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (2000 mg, 9.47 mmol) in THF (5 mL) was added dropwise at 0°C. The resulting mixture was gradually heated to 25°C and stirred for a further 2 hours. LC-MS showed that the starting material was consumed and the desired product was formed. The reaction mixture was quenched with water (50 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with brine (150 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (Biotage, 80g silica gel, 0%-30% phenylethyl in petroleum ether) to obtain crude ethyl(S)-2-(2-(tert-butoxy)-2-oxoethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (1870mg, crude product) as a colorless oil. LCMS (ESI) m / z: 310.1 [M+H]+.

[0411] Step 2: Synthesis of (S)-2-(7a-(ethoxycarbonyl)-5-oxotetrahydro-1H-pyrrolidine-2(3H)-ylidene)acetic acid (I-8b) To a solution of ethyl(S)-2-(2-(tert-butoxy)-2-oxoethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (700 mg, 2.26 mmol) in DCM (8 mL), HCl (6 mL, 4.0 M in dioxane) was added. The mixture was then stirred at 25°C for 16 hours. LC-MS showed that the starting material was consumed and the desired product was formed. The solution was concentrated under reduced pressure to obtain (S)-2-(7a-(ethoxycarbonyl)-5-oxotetrahydro-1H-pyrrolidine-2(3H)-ylidene)acetic acid (550 mg, crude) as a pale yellow oil. LC-MS (ESI) m / z: 254.1 [M+H] + .

[0412] Step 3: Synthesis of ethyl(S)-2-(2-hydroxyethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate(I-8c) To a solution of (S)-2-(7a-(ethoxycarbonyl)-5-oxotetrahydro-1H-pyrrolidine-2(3H)-ylidene)acetic acid (970 mg, 3.83 mmol) and NMM (774 mg, 7.65 mmol) in THF (33 mL), isobutyl chloroformate (784 mg, 5.74 mmol) in THF (4 mL) was added dropwise at 0°C. The mixture was stirred at 0°C for 40 minutes. A precipitate formed, and LC-MS indicated the formation of a mixed anhydrous intermediate. The solid was filtered. NaBH4 (247 mg, 6.53 mmol) in water (5 mL) was added to the filtrate at 0°C. The resulting mixture was stirred at 0°C for 15 minutes. LC-MS indicated the formation of the desired product. The solution was concentrated under reduced pressure. The residue was purified by flash column chromatography (Biotage, 25 g silica gel, 0-100% ethyl in petroleum ether) to obtain ethyl(S)-2-(2-hydroxyethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (220 mg, 24%, E / Z mixture) as a colorless oil. 1 H NMR (400 MHz, CDCl3) δ 5.71 - 5.57 (m, 1H), 4.33 (d, J = 16.0 Hz, 1H), 4.21 (q, J = 7.1 Hz, 2H), 4.16 - 4.08 (m, 2H), 3.78 (d, J = 17.0 Hz, 1H), 3.24 (d, J = 15.7 Hz, 0.2H, minor isomer), 3.07 (d, J = 15.8 Hz, 0.8H, main isomer), 2.84 - 2.73 (m, 1H), 2.67 - 2.57 (m, 1H), 2.55 - 2.37 (m, 2H), 2.18 - 2.06 (m, 1H), 1.28 (t, J = 7.1 Hz, 3H). LCMS (ESI) m / z 240.1 [M+H] + .

[0413] Step 4: Synthesis of ethyl(S)-2-(2-methoxyethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate(I-8d) Iodomethane (1070 mg, 7.52 mmol) was added to a solution of ethyl(S)-2-(2-hydroxyethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (180 mg, 0.752 mmol) and Ag2O (872 mg, 3.76 mmol) in CH3CN (10 mL). The reaction mixture was stirred at 25°C for 16 hours. LC-MS showed that approximately 50% of the starting material remained and approximately 50% of the desired product was formed. Another portion of Ag2O (436 mg, 1.88 mmol) was added, and the mixture was stirred for a further 16 hours. LC-MS showed that the starting material was consumed and the desired product was formed. The mixture was filtered. The filtrate was concentrated under reduced pressure to obtain crude ethyl(S)-2-(2-methoxyethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (200 mg, crude product, E / Z mixture) as a pale yellow solid. 1 H NMR (400 MHz, CDCl3) δ 5.54 - 5.45 (m, 1H), 4.31 - 4.21 (m, 1H), 4.13 (q, J = 7.1 Hz, 2H), 3.86 - 3.77 (m, 2H), 3.71 (d, J = 15.6 Hz, 1H), 3.24 (s, 3H), 3.15 (d, J = 16.7 Hz, 0.2H, minor isomer), 3.00 (d, J = 15.8 Hz, 0.8H, main isomer), 2.79 - 2.66 (m, 1H), 2.61 - 2.50 (m, 1H), 2.49 - 2.33 (m, 2H), 2.11 - 2.00 (m, 1H), 1.20 (t, J = 7.1 Hz, 3H). LCMS (ESI) m / z: 254.1 [M+H] + .

[0414] Step 5: Separation of ethyl(S,Z)-2-(2-methoxyethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate(I-8e) An E / Z mixture of ethyl(S)-2-(2-methoxyethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (200 mg, 0.79 mmol) was separated by chiral SFC (instrument: SFC 150; column: Daicel CHIRALPAK IC, 250 mm × 30 mm ID, 10 μm; mobile phase: CO2 / MeOH [0.2% NH3 (7M solution in MeOH)] = 90 / 10; flow rate: 120 g / min; wavelength: UV 214 nm; temperature: 35 °C) to obtain ethyl(S,Z)-2-(2-methoxyethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (91 mg, 46%, Rt: 2.406 min, peak 1) as a pale yellow oil. 1 H NMR (400 MHz, CDCl3) δ 5.63 - 5.52 (m, 1H), 4.32 (d, J = 16.0 Hz, 1H), 4.20 (q, J = 7.1 Hz, 2H), 3.87 (d, J = 6.5 Hz, 2H), 3.78 (d, J = 15.9 Hz, 1H), 3.31 (s, 3H), 3.07 (d, J = 15.8 Hz, 1H), 2.78 (dt, J = 16.7, 9.7 Hz, 1H), 2.61 (ddd, J = 13.2, 9.1, 1.8 Hz, 1H), 2.56 - 2.40 (m, 2H), 2.18 - 2.05 (m, 1H), 1.27 (t, J = 7.1 Hz, 3H). LCMS (ESI) m / z: 254.1 [M+H] + .

[0415] Step 6: Synthesis of (S,Z)-(2-(2-methoxyethylidene)tetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol (Compound I-8) To a solution of ethyl(S,Z)-2-(2-methoxyethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (80 mg, 0.32 mmol) in THF (3.2 mL), LAH (0.63 mL, 1 M in THF) was added under nitrogen at 0°C. The resulting mixture was heated to 70°C and stirred at 70°C for 1 hour. TLC (DCM / MeOH = 10:1, iodine staining) showed that the starting material had been consumed and new points were observed. The mixture was quenched with Na2SO4.10H2O and the suspension was stirred at 25°C for 2 hours. The mixture was filtered and the filtrate was concentrated to obtain crude (S,Z)-(2-(2-methoxyethylidene)tetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol (58 mg, 93%) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 5.52 - 5.42 (m, 1H), 3.88 - 3.81 (m, 2H), 3.68 (d, J = 15.6 Hz, 1H), 3.36 - 3.21 (m, 6H), 3.15 - 3.05 (m, 1H), 2.67 (dt, J = 10.1, 7.1 Hz, 1H), 2.50 (d, J = 16.1 Hz, 1H), 2.38 (d, J = 16.3 Hz, 1H), 1.93 - 1.78 (m, 3H), 1.72 - 1.65 (m, 1H). LCMS (ESI) m / z 198.3 [M+H] + . (Note: Separately, E isomers were also prepared from pure E-esters by LAH reduction using a similar method.)

[0416] Intermediate 9(I9-i and I9-ii): (5R)-4-bromo-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-5,6,7,8-tetrahydro-1H-benzo[f]indazole and (5S)-4-bromo-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-5,6,7,8-tetrahydro-1H-benzo[f]indazole

[0417] [ka]

[0418] Step 1: Synthesis of (E)-6-fluoro-3,4-dihydronaphthalene-1(2H)-one O-methyloxime (I-9a) A solution of 6-fluoro-1-tetralone (4.00 g, 24.4 mmol), methoxylamine hydrochloride (3.05 g, 36.5 mmol), and pyridine (2.89 g, 36.5 mmol) in CH3CN (20 mL) was stirred at 25°C for 2 hours. LC-MS showed that the desired product was formed. The reaction mixture was diluted with water (100 mL) and extracted with ELISA (100 mL x 3). The organic layers were combined, washed with 2 M aqueous HCl (100 mL x 2) and brine (200 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain (E)-6-fluoro-3,4-dihydronaphthalene-1(2H)-one O-methyloxime (4.90 g, crude) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 7.96 (dd, J = 8.8, 6.0 Hz, 1H), 6.88 (td, J = 8.6, 2.7 Hz, 1H), 6.82 (dd, J = 9.3, 2.6 Hz, 1H), 3.97 (s, 3H), 2.76 - 2.67 (m, 4H), 1.88 - 1.77 (m, 2H). LCMS (ESI) m / z: 194.1 [M+H] + .

[0419] Step 2: Synthesis of (E)-8-bromo-6-fluoro-3,4-dihydronaphthalene-1(2H)-one O-methyloxime (I-9b) Palladium acetate (422 mg, 2.54 mmol) was added under nitrogen to a mixture of (E)-6-fluoro-3,4-dihydronaphthalene-1(2H)-one O-methyloxime (4.90 g, 25.4 mmol) and NBS (5.42 g, 30.4 mmol) in acetic acid (121 mL). The reaction mixture was then stirred at 90°C for 3 hours. LC-MS showed that the starting materials had been consumed and the desired compound was found. The mixture was filtered through a Celite pad, and the filtrate was concentrated to obtain the crude product. The crude product was purified by flash column chromatography (Combi-Flash, 80 g silica gel, 0-5% ethyl phosphate in petroleum ether) to obtain (E)-8-bromo-6-fluoro-3,4-dihydronaphthalene-1(2H)-one O-methyloxime (5.60 g, 81%) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 7.27 (dd, J = 8.5, 2.7 Hz, 1H), 6.84 (dd, J = 8.3, 2.6 Hz, 1H), 4.02 (s, 3H), 2.75 (t, J = 6.9 Hz, 2H), 2.65 - 2.57 (m, 2H), 1.81 - 1.70 (m, 2H). LCMS (ESI) m / z: 272.1, 274.0 [M+H] + .

[0420] Step 3: Synthesis of 8-bromo-6-fluoro-3,4-dihydronaphthalene-1(2H)-one (I-9c) To a solution of (E)-8-bromo-6-fluoro-3,4-dihydronaphthalene-1(2H)-one O-methyloxime (5.00 g, 18.3 mmol) in 1,4-dioxane (80 mL), dilute sulfuric acid (92 mL, 4 M) was added at 25 °C. The mixture was stirred at 110 °C for 20 hours. LC-MS showed that the starting material was consumed and the desired product was formed. The reaction mixture was made basic to pH 8 with aqueous NaOH (1 M) and extracted with ethyl acetate (100 mL). The organic layers were combined, washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by flash column chromatography (Combi-Flash, 80 g silica gel, 0-5% gradient of siRNA in petroleum ether) to obtain 8-bromo-6-fluoro-3,4-dihydronaphthalene-1(2H)-one (3.00 g, 67%) as a yellow oily substance. 1 H NMR (400 MHz, CDCl3) δ 7.31 (dd, J = 8.3, 2.5 Hz, 1H), 6.96 - 6.91 (m, 1H), 2.98 (t, J = 6.2 Hz, 2H), 2.72 - 2.65 (m, 2H), 2.15 - 2.06 (m, 2H). LCMS (ESI) m / z: 243.0, 245.0 [M+H] + .

[0421] Step 4: Synthesis of 8-bromo-6-fluoro-1-methyl-1,2,3,4-tetrahydronaphthalene (I-9d) A solution of MeMgBr in THF (3M, 12.3 mL, 37.0 mmol) was added dropwise at 0°C to a solution of 8-bromo-6-fluoro-3,4-dihydronaphthalene-1(2H)-one (3.00 g, 12.3 mmol) in THF (50 mL). After the addition was complete, the solution was stirred at 25°C for a further 30 minutes. LC-MS showed that most of the starting material had been consumed and an alcohol intermediate was detected. The reaction mixture was quenched with saturated aqueous NH4Cl (25 mL). The mixture was extracted with ELISA (100 mL). The combined organic extract was dried over MgSO4 and concentrated. The residue was dissolved in DCM (50 mL). Triethylsilane (5.01 g, 43.2 mmol) and trifluoroacetic acid (3.3 mL) were added at -60°C. The reaction mixture was stirred at 25°C for 3 hours. LC-MS indicated that the intermediate was consumed and the desired MS was detected. The reaction mixture was quenched by adding saturated NaHCO3 aqueous solution and extracted with DCM (100 mL). The combined organic extract was dried over MgSO4 and concentrated under vacuum. The residue was purified by flash column chromatography (Combi-Flash, 80 g silica gel, 0-2% gradient of siRNA in petroleum ether) to obtain 8-bromo-6-fluoro-1-methyl-1,2,3,4-tetrahydronaphthalene (2.25 g, 75%) as a colorless oil. 1 H NMR (400 MHz, CDCl3) δ 7.12 (dd, J = 8.1, 2.6 Hz, 1H), 6.77 - 6.72 (m, 1H), 3.23 - 3.14 (m, 1H), 2.84 - 2.67 (m, 2H), 1.91 - 1.72 (m, 4H), 1.21 (d, J = 7.0 Hz, 3H).

[0422] Step 5: Synthesis of 1-bromo-3-fluoro-8-methyl-5,6,7,8-tetrahydronaphthalene-2-carboaldehyde (I-9e) A solution of 2,2,6,6-tetramethylpiperidine (2.09 g, 14.8 mmol) in THF (20 mL) was cooled to -78°C. Then, n-BuLi (2.5 M, 4.3 mL, 10.9 mmol) was added to the above solution. The reaction mixture was stirred at -60°C for 30 minutes. Then, a solution of 8-bromo-6-fluoro-1-methyl-1,2,3,4-tetrahydronaphthalene (1.2 g, 4.9 mmol) in THF (20 mL) was added to the above solution, and stirring was continued at -60°C for 30 minutes. Then, DMF (1.1 mL, 14.8 mmol) was added to the above solution at -60°C. The mixture was warmed to 15°C and stirred at 15°C for 3 hours. LC-MS showed that the starting material was consumed and the desired MS was detected. The mixture was quenched with saturated NH4Cl aqueous solution (50 mL) and extracted with Â1 (50 mL x 2). The combined organic layer was washed with brine (50 mL), dried, and concentrated to obtain the crude product. The crude product was combined with another batch and purified by flash column chromatography (Combi-Flash, 12 g silica gel, 0-15% petroleum ether in DCM) to obtain 1-bromo-3-fluoro-8-methyl-5,6,7,8-tetrahydronaphthalene-2-carboaldehyde (900 mg, 40%) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 10.38 (s, 1H), 6.86 (d, J = 11.0 Hz, 1H), 3.41 - 3.32 (m, 1H), 2.93 - 2.73 (m, 2H), 1.96 - 1.76 (m, 4H), 1.24 (d, J = 7.0 Hz, 3H). LCMS (ESI) m / z: 271.1, 273.0 [M+H] + .

[0423] Step 6: Synthesis of 4-bromo-5-methyl-5,6,7,8-tetrahydro-1H-benzo[f]indazole (I-9f) To a mixture of 1-bromo-3-fluoro-8-methyl-5,6,7,8-tetrahydronaphthalene-2-carboaldehyde (700 mg, 2.58 mmol) in ethylene glycol (10 mL), hydrazine hydrate (1.62 g, 25.8 mmol) was added at 10 °C, and the mixture was then stirred at 130 °C for 48 hours. LC-MS showed that the starting material was consumed and the desired product was formed. Water (10 mL) was added. The aqueous phase was extracted with toluene (20 mL), dried over Na₂SO₄, and filtered. The solvent was evaporated, and the residue was purified by flash column chromatography (Combi-Flash, silica gel 4 g, petroleum ether / toluene = 90:10) to obtain 4-bromo-5-methyl-5,6,7,8-tetrahydro-1H-benzo[f]indazole (330 mg, 48%) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 8.02 (s, 1H), 7.16 (s, 1H), 3.48 - 3.40 (m, 1H), 3.04 - 2.88 (m, 2H), 1.90 - 1.80 (m, 4H), 1.28 (d, J = 7.0 Hz, 3H). LCMS (ESI) m / z: 265.1, 267.0 [M+H] + .

[0424] Step 7: Chiral separation of 4-bromo-5-methyl-5,6,7,8-tetrahydro-1H-benzo[f]indazole (to obtain I-9f-i and I-9f-ii) Racemic 4-bromo-5-methyl-5,6,7,8-tetrahydro-1H-benzo[f]indazole (300 mg, 1.13 mmol) was separated by chiral SFC (instrument: SFC 80; column: Daicel CHIRALCEL AD, 250 mm × 30 mm ID, 10 μm; mobile phase: CO2 / MeOH [0.2% NH3 (7 M solution in MeOH)] = 80 / 20; flow rate: 70 g / min; wavelength: UV 214 nm; temperature: 35 °C) to obtain two isomers: (R)-4-bromo-5-methyl-5,6,7,8-tetrahydro-1H-benzo[f]indazole (I9f-i) (90 mg, 30%, Rt: 1.487 mins, peak 1) as a white solid. 1 H NMR (400 MHz, CDCl3) δ 8.03 (s, 1H), 7.18 (s, 1H), 3.50 - 3.40 (m, 1H), 3.04 - 2.84 (m, 2H), 2.01 - 1.74 (m, 4H), 1.28 (d, J = 7.0 Hz, 3H). LCMS (ESI) m / z: 265.0, 267.0 [M+H] + .[α] D 25 = -29.09 (c 0.11, MeOH). This isomer was taken up to obtain Example 29. (S)-4-bromo-5-methyl-5,6,7,8-tetrahydro-1H-benzo[f]indazole (I9f-ii) (80 mg, 27%, Rt: 1.860 min, peak 2) was taken as a white solid. 1 H NMR (400 MHz, CDCl3) δ 8.01 (s, 1H), 7.16 (s, 1H), 3.48 - 3.39 (m, 1H), 3.04 - 2.82 (m, 2H), 1.98 - 1.79 (m, 4H), 1.27 (d, J = 7.0 Hz, 3H). LCMS (ESI) m / z: 265.0, 266.9 [M+H] + .[α] D 25 = +17.00 (c 0.1, MeOH). This isomer was selected to obtain Example 32.

[0425] Step 8: Synthesis of (5R)-4-bromo-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-5,6,7,8-tetrahydro-1H-benzo[f]indazole (I-9-i) To a solution of (R)-4-bromo-5-methyl-5,6,7,8-tetrahydro-1H-benzo[f]indazole (I-9f-i) (50 mg, 0.19 mmol) in THF (3.0 mL), DHP (47 mg, 0.56 mmol), followed by p-toluenesulfonic acid monohydrate (36 mg, 0.19 mmol), was added. The reaction mixture was stirred at 60°C for 16 hours. LC-MS showed that the desired product had formed. The mixture was cooled and washed with saturated NaHCO3 aqueous solution (10 mL) and brine (10 mL). The organic layer was dried and concentrated. The residue was combined with the crude product from another batch and purified by flash column chromatography (Combi-Flash, 4 g silica gel, siRNA in petroleum ether from 0 to 10%) to obtain (5R)-4-bromo-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-5,6,7,8-tetrahydro-1H-benzo[f]indazole (70 mg, 89%) as a pale yellow oil. 1 H NMR (400 MHz, CDCl3) δ 7.96 (s, 1H), 7.25 (s, 1H), 5.63 (dt, J = 9.3, 2.8 Hz, 1H), 4.04 - 3.94 (m, 1H), 3.77 - 3.69 (m, 1H), 3.54 - 3.41 (m, 1H), 3.07 - 2.88 (m, 2H), 2.60 - 2.46 (m, 1H), 2.20 - 1.75 (m, 9H), 1.26 - 1.24 (m, 3H). LCMS (ESI) m / z: 349.1, 351.1 [M+H] + .

[0426] Step 9: Synthesis of (5S)-4-bromo-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-5,6,7,8-tetrahydro-1H-benzo[f]indazole (I-9-ii) To a solution of (S)-4-bromo-5-methyl-5,6,7,8-tetrahydro-1H-benzo[f]indazole (I9f-ii) (80 mg 0.30 mmol) in THF (4.0 mL), DHP (76 mg 0.905 mmol), followed by p-toluenesulfonic acid monohydrate (57 mg 0.302 mmol), was added. The reaction mixture was stirred at 60°C for 16 hours. LC-MS showed that the desired product had formed. The mixture was cooled and washed with saturated NaHCO3 aqueous solution (10 mL) and brine (10 mL). The organic layer was dried, concentrated, and the residue was purified by flash column chromatography (Combi-Flash, silica gel 4 g, ethyl ether in petroleum ether from 0 to 10%) to obtain (5S * )-4-bromo-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-5,6,7,8-tetrahydro-1Hbenzo[f]indazole) (72 mg, 68%) was obtained as a pale yellow oily substance. 1 H NMR (400 MHz, CDCl3) δ 7.96 (s, 1H), 7.25 (s, 1H), 5.64 (dt, J = 9.3, 2.9 Hz, 1H), 4.04 - 3.99 (m, 1H), 3.76 - 3.69 (m, 1H), 3.46 - 3.39 (m, 1H), 3.04 - 2.91 (m, 2H), 2.59 - 2.45 (m, 1H), 2.18 - 1.77 (m, 9H), 1.26 - 1.24 (m, 3H). LCMS (ESI) m / z: 349.0, 351.0 [M+H] + .

[0427] Intermediate 10(I-10):rac-4-bromo-6-chloro-5-(cis-2-methylcyclopropyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole

[0428] [ka]

[0429] Step 1: Synthesis of 3-bromo-5-chloro-4-iodo-2-methylaniline (I-10a) To a stirred solution of 3-bromo-5-chloro-2-methylaniline (10 g, 45 mmol) in CH3CN (113 mL), indium(III) trifluoromethanesulfonate (12.7 g, 22.7 mmol), followed by iodine monochloride (7.36 g, 45.4 mmol), was added dropwise at 0°C. The reaction mixture was stirred at 0°C for 2 hours. Then, it was heated to 25°C and stirred at 25°C for 16 hours. LC-MS showed that most of the starting material had been consumed and the desired product had been formed. The reaction mixture was quenched with triethylamine (9.5 mL, 68 mmol) and then stirred for 15 minutes. The solid was removed by filtration through Celite. The filtrate cake was rinsed with siRNA (100 mL). The filtrate was collected, washed with brine (70 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (120 g silica gel, 0-5% siRNA in petroleum ether) to obtain 3-bromo-5-chloro-4-iodo-2-methylaniline (9.5 g, 60%) as a solid. 1 H NMR (400 MHz, CDCl3) δ 6.85 (s, 1H), 3.82 (s, 2H), 2.38 (s, 3H). LCMS (ESI) m / z: 345.9, 347.8 [M+H] + .

[0430] Step 2: Synthesis of 3-bromo-5-chloro-4-iodo-2-methylbenzenediazonium tetrafluoroborate (I-10b) Diethyl boron trifluoride etherate (2.46 g, 17.3 mmol) was dissolved in DCM (35 mL) and cooled to -5 to -10°C under a nitrogen atmosphere. A solution of 3-bromo-5-chloro-4-iodo-2-methylaniline (4 g, 11.6 mmol) in DCM (5 mL) was added to the above reaction mixture and stirred for 0.5 hours. Then, tert-butyl nitrite (1.79 g, 17.3 mmol) was added dropwise, and the reaction mixture was stirred at -5 to -10°C for 1.5 hours. TLC (petroleum ether / ethyl ether = 5:1, UV 254 nm) showed that the starting material (Rf = 0.15) was completely consumed. MTBE (50 mL) was added to the reaction mixture to form a precipitate, which was then filtered under vacuum and washed with cold MTBE (10 mL x 2) to obtain 3-bromo-5-chloro-4-iodo-2-methylbenzenediazonium tetrafluoroborate (4.6 g, 90%) as a white solid. 1 H NMR (400 MHz, MeOD) δ 8.79 (s, 1H), 2.94 (s, 3H).

[0431] Step 3: Synthesis of 4-bromo-6-chloro-5-iodo-1H-indazole (I-10c) To a solution of 18-crown-6 (819 mg, 3.1 mmol) in chloroform (50 mL), KOAc (1.42 g, 14.5 mmol) was added, and the mixture was cooled to 0°C. Then, 3-bromo-5-chloro-4-iodo-2-methylbenzenediazonium tetrafluoroborate (4.6 g, 10.3 mmol) was slowly added. The reaction mixture was then stirred at 25°C for 2 hours. LC-MS showed that the desired product had formed. The reaction mixture was poured into ice-cold water (100 mL) and extracted with DCM (50 mL x 3). The combined organic layers were washed with saturated NaHCO3 aqueous solution (50 mL) and brine (50 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by flash column chromatography (40 g silica gel, 9% siRNA in petroleum ether) to obtain 4-bromo-6-chloro-5-iodo-1H-indazole (1.95 g, 53%) as a yellow solid. 1H NMR (400 MHz, CDCl3) δ 8.02 (s, 1H), 7.68 (d, J = 0.9 Hz, 1H). LCMS (ESI) m / z: 356.8, 358.7 [M+H]+.

[0432] Step 4: Synthesis of 4-bromo-6-chloro-5-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole (I-10d) To a solution of 4-bromo-6-chloro-5-iodo-1H-indazole (1.95 g, 5.45 mmol) in DCM (50 mL), DHP (688 mg, 8.18 mmol), followed by p-toluenesulfonic acid monohydrate (104 mg, 0.546 mmol), the reaction mixture was stirred at 30°C for 16 hours. LC-MS showed that the starting material had been consumed. The mixture was successively washed with saturated NaHCO3 aqueous solution (30 mL) and brine (30 mL). The organic layer was dried, concentrated, and purified by flash column chromatography (12 g silica gel, ethyl phosphate in 0-5% PE) to obtain 4-bromo-6-chloro-5-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole (2.1 g, 87%) as a yellow solid. 1 H NMR (400 MHz, CDCl3) δ 7.92 (s, 1H), 7.78 (d, J = 0.7 Hz, 1H), 5.65 (dd, J = 8.9, 2.7 Hz, 1H), 4.04 - 3.92 (m, 1H), 3.84 - 3.62 (m, 1H), 2.55 - 2.38 (m, 1H), 2.16 - 2.02 (m, 2H), 1.80 - 1.67 (m, 3H). LCMS (ESI) m / z: 356.8 and 358.9 [M-THP+H]+.

[0433] Step 5: Synthesis of rac-4-bromo-6-chloro-5-((1R,2S)-2-methylcyclopropyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole(I-10) Pd(dppf)Cl2 (36 mg, 0.05 mmol) was added to a mixture of 4-bromo-6-chloro-5-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole (217 mg, 0.49 mmol), tripotassium phosphate (209 mg, 0.98 mmol), and rac-cis-4,4,5,5-tetramethyl-2-(cis-2-methylcyclopropyl)-1,3,2-dioxaborolane (268 mg, 1.47 mmol) in 1,4-dioxane (5 mL) and water (0.5 mL). The mixture was degassed with nitrogen for 2 minutes and then stirred at 110°C for 16 hours. Three other reactions were set up in parallel on the same scale. LC-MS showed that the starting materials were consumed and approximately 25% of the desired product was detected (UV 214 nm). The mixtures of all four reactions were combined, diluted with water (100 mL), and extracted with siRNA (50 mL x 3). The combined organic layer was dried over Na₂SO₄ and concentrated under reduced pressure. The residue was purified by flash column chromatography (elution with 3% siRNA in 12 g silica gel and petroleum ether), followed by flash reversed-phase column chromatography (elution with a gradient of 0-70% CH₃CN in 40 g C₁₄ water (0.1% FA)) to obtain rac-4-bromo-6-chloro-5-(cis)-2-methylcyclopropyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole (200 mg, 27%) as an oil. 1 ¹H NMR (400 MHz, CDCl3) δ 7.98 (s, 1H), 7.64 (s, 1H), 5.63 and 5.62 (2 dd, J = 9.2, 2.7 Hz, 1H, (main diastereomer and secondary diastereomer, respectively), 4.08 - 3.93 (m, 1H), 3.81 - 3.66 (m, 1H), 2.59 - 2.40 (m, 1H), 2.18 - 2.03 (m, 2H), 1.95 (td, J = 8.5, 6.7 Hz, 1H), 1.82 - 1.63 (m, 3H), 1.46 - 1.32 (m, 2H), 1.31 - 1.21 (m, 1H), 0.89 - 0.84 (m, 1H), 0.81 and 0.81 (2 d, J = 6.0 Hz, 3H (primary and secondary diastereomers, respectively). LCMS (ESI) m / z: 369.0, 370.9 [M+H]+. Note: This intermediate was incorporated into the final product according to Scheme A. Separation of the final analogue was achieved by chiral-HPLC (column: CHIRALPAK IK 2.5cm ID × 25cm L, 10 μm; mobile phase: CH3CN / DEA = 100 / 0.1 (V / V); flow rate: 30 mL / min), followed by preparative HPLC purification to obtain Examples 30 and 31.

[0434] Intermediate 11(I-11):4-bromo-6-chloro-5-ethyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole

[0435] [ka]

[0436] Step 1: Synthesis of 4-bromo-6-chloro-1-(tetrahydro-2H-pyran-2-yl)-5-vinyl-1H-indazole (I-11a) To a solution of 4-bromo-6-chloro-5-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole (450 mg, 1.02 mmol) and 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (314 mg, 2.04 mmol) in 1,4-dioxane (10 mL) and water (1 mL), Pd(dppf)Cl2 (75 mg, 0.102 mmol) and tripotassium phosphate (649 mg, 3.06 mmol) were added. The mixture was then degassed with nitrogen and stirred at 60°C for 2 hours. LC-MS showed that the starting material was consumed and the desired product was formed. The reaction mixture was poured into  (50 mL) and the layers were partitioned. The organic layer was washed with water (30 mL) and dried over Na2SO4. After filtration, the filtrate was concentrated under vacuum. The residue was purified by flash column chromatography (Combi-Flash, 12g silica gel, 0-10% siRNA in petroleum ether) to obtain 4-bromo-6-chloro-1-(tetrahydro-2H-pyran-2-yl)-5-vinyl-1H-indazole (220mg, 63%) as a white solid. LCMS (ESI) m / z: 340.9, 343.0 [M+H] + .

[0437] Step 2: Synthesis of 4-bromo-6-chloro-5-ethyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole (I-11) To a solution of 4-bromo-6-chloro-1-(tetrahydro-2H-pyran-2-yl)-5-vinyl-1H-indazole (220 mg, 0.644 mmol) in  (5 mL) and isopropanol (5 mL), platinum(IV) oxide (146 mg, 0.644 mmol) was added. The reaction mixture was then stirred at 45 °C under a hydrogen (15 psi) atmosphere for 3 hours. LC-MS showed that the starting material was consumed and the desired product was formed. The mixture was filtered, and the filter cake was washed with  (30 mL). The filtrate was concentrated under vacuum. The residue was purified by preparative TLC ( / petroleum ether = 1 / 15, UV 254 nm) to obtain 4-bromo-6-chloro-5-ethyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole (80 mg, 36%) as a white solid. 1 H NMR (400 MHz, CDCl3) δ 7.89 (d, J = 3.0 Hz, 1H), 7.56 (d, J = 3.0 Hz, 1H), 5.61 - 5.53 (m, 1H), 3.93 (d, J = 8.2 Hz, 1H), 3.67 (t, J = 10.0 Hz, 1H), 3.00 (dt, J = 14.7, 5.4 Hz, 2H), 2.41 (dd, J = 11.1, 8.5 Hz, 1H), 2.07 (s, 1H), 1.77 - 1.60 (m, 4H), 1.12 (td, J = 7.4, 3.8 Hz, 3H). LCMS (ESI) m / z: 343.0, 345.0 [M+H] + .

[0438] Intermediate 12(I-12):(8aS,13S)-5-chloro-4-fluoro-13-methyl-2-(methylthio)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene

[0439] [ka]

[0440] Step 1: Synthesis of (S)-3-(benzylamino)butan-1-ol (I-12a) Benzaldehyde (2.10 g, 2.01 mL, 1 equivalent, 19.8 mmol) was added to a solution of (3S)-3-aminobutan-1-ol (1.765 g, 1.90 mL, 1.00 equivalent, 19.80 mmol) in methanol (30 mL). The mixture was stirred at room temperature for 1 hour. The reaction mixture was then cooled in an ice bath, and sodium borohydride (749 mg, 1 equivalent, 19.8 mmol) was added gradually while maintaining controlled effervescence. After 20 minutes, the reaction mixture was removed from the ice bath and stirred at room temperature for 2 hours. LC-MS showed the desired mass and no residual benzaldehyde peak. Water was added, and the reaction mixture was concentrated under vacuum. The white solid was placed in water (40 mL) and extracted with DCM (2 × 40 mL). The combined DCM layers were dried on Na2SO4, filtered, and concentrated to obtain (S)-3-(benzylamino)butan-1-ol (3.41 g, 96%) as a clear oil. 1 ¹H NMR (400 MHz, methanol-d4) δ = 7.38 - 7.28 (m, 4H), 7.28 - 7.21 (m, 1H), 3.84 - 3.79 (m, 1H), 3.75 - 3.58 (m, 3H), 2.87 (sxt, J = 6.4 Hz, 1H), 1.76 (tdd, J = 6.2, 7.8, 14.0 Hz, 1H), 1.56 (qd, J = 5.8, 14.0 Hz, 1H), 1.13 (d, J = 6.3 Hz, 3H). LCMS (ES-API) m / z: 180.1 [M+H] + .

[0441] Step 2: Synthesis of (S)-3-(benzyl(oxetan-3-yl)amino)butan-1-ol(I-12b) A solution of (S)-3-(benzylamino)butan-1-ol (3.28 g, 1 equivalent, 18.3 mmol) in toluene (30 mL) was treated with oxetane-3-one (1.52 g, 1.35 mL, 1.15 equivalents, 21.1 mmol) and heated to 105 °C for 3 hours. The reaction mixture was cooled to 35 °C and sodium triacetoxyborohydride (6.21 g, 1.60 equivalents, 29.3 mmol) was added. The reaction was continued overnight at 35 °C. The reaction mixture was quenched with 2N Na₂CO₃ (40 mL) and vigorously stirred at room temperature for 1 hour. The layers were separated. The aqueous layer was extracted again with toluene. The combined toluene layers were concentrated under vacuum at 40°C to obtain (S)-3-(benzyl(oxetan-3-yl)amino)butan-1-ol (4.14 g, 96%) as a transparent oil. 1 ¹H NMR (400 MHz, methanol-d4) δ = 7.37 - 7.33 (m, 2H), 7.32 - 7.26 (m, 2H), 7.25 - 7.20 (m, 1H), 4.68 - 4.63 (m, 1H), 4.56 (t, J = 6.6 Hz,1H), 4.45 - 4.35 (m, 2H), 4.30 - 4.20 (m, 1H), 3.92 (d, J = 14.0 Hz, 1H), 3.66 - 3.52 (m, 3H), 2.89 - 2.78 (m, 1H), 1.79 (qd, J = 7.1, 14.5 Hz, 1H),1.41 (qd, J = 6.1, 13.9 Hz, 1H), 0.99 (d, J = 6.6 Hz, 3H). LCMS (APCI) m / z: 236.1 [M+H] + .

[0442] Step 3: Synthesis of ((3R,5S)-4-benzyl-5-methyl-1,4-oxazepan-3-yl)methanol (I-12c-ii) To a solution of (S)-3-(benzyl(oxetan-3-yl)amino)butan-1-ol (3.916 g, 1 equivalent, 16.64 mmol) in toluene (40 mL), dibutyl hydrogen phosphate (3.8 g, 3.5 mL, 1.1 equivalents, 18 mmol) was added. The reaction mixture was heated to 108 °C (oil bath temperature) for 20 hours. The reaction mixture was cooled to room temperature, and 2N Na₂CO₃ (40 mL) was added. The mixture was vigorously stirred for 1 hour. The layers were separated, and the toluene layer was concentrated under vacuum. Crude 1 The 1H NMR spectrum shows a 2.1:1 mixture of diastereoisomers as a pale yellow oily substance (3.61 g, 15.3 mmol, 92%).

[0443] Diastereoisomers were separated by chiral SFC (column: Chiralpak IG SFC 5um 30mm × 250mm, mobile phase A: CO2, mobile phase B: MeOH + 10mm NH3 12%B fixed composition, 120 bar, 120 mL / min) to ((3S,5S)-4-benzyl-5-methyl-1,4-oxazepan-3-yl)methanol (1.53g, 6.50 mmol, 39%) (I-12c-i, trans product) >99.0% de, [a]D22 = -70.7° (C 0.1, MeOH). 1 ¹H NMR (400 MHz, methanol-d4) δ = 7.38 - 7.33 (m, 2H), 7.32 - 7.25 (m, 2H), 7.23 - 7.17 (m, 1H), 4.06 (dd, J = 5.0, 12.9 Hz, 1H), 4.00 - 3.90 (m, 2H), 3.70 (d, J = 14.0 Hz, 1H), 3.67 - 3.59 (m, 2H), 3.43 - 3.33 (m, 2H), 2.94 (dq, J = 5.1, 7.5 Hz, 1H), 1.97 (dtd, J = 5.3, 9.3, 14.7 Hz, 1H), 1.79 - 1.70 (m, 1H), 1.24 (d, J = 6.9 Hz, 3H). The 1H peak is hidden by the solvent peak. LCMS (APCI): 236.1 [M+H]+ And ((3R,5S)-4-benzyl-5-methyl-1,4-oxazepan-3-yl)methanol (0.739 g, 3.14 mmol, 19%) (I-12c-ii, cis product) >99.0% de, [a]D22 = -14.8° (C 0.1, MeOH) was obtained. 1 ¹H NMR (400 MHz, methanol-d4) δ = 7.39 (d, J = 7.6 Hz) 2H), 7.30 - 7.23 (m, 2H), 7.20 - 7.13 (m, 1H), 3.90 - 3.76 (m, 5H), 3.68 (ddd, J = 2.6, 9.8, 12.6 Hz, 1H), 3.40 - 3.34 (m, 1H), 3.29 - 3.21 (m, 2H), 3.20 - 3.12 (m, 1H), 1.97 - 1.86 (m, 1H), 1.75 (qdd, J = 2.6, 3.5, 15.0 Hz, 1H), 1.12 (d, J = 6.6 Hz, 3H). LCMS (APCI): 236.1 [M+H] + .

[0444] Step 4: Synthesis of ((3R,5S)-5-methyl-1,4-oxazepan-3-yl)methanol (I-12d) To a solution of ((3R,5S)-4-benzyl-5-methyl-1,4-oxazepan-3-yl)methanol (737 mg, 1 equivalent, 3.13 mmol) in methanol (20 mL), palladium(II) dihydrate (44.0 mg, 0.1 equivalent, 313 μmol) was added. The reaction mixture was vigorously stirred under H2 for 18 hours. The reaction mixture was filtered and concentrated under vacuum. The resulting clear oily substance, ((3R,5S)-5-methyl-1,4-oxazepan-3-yl)methanol (433 mg, 2.98 mmol, 95%), was used without further purification. 1 ¹H NMR (400 MHz, methanol-d4) δ = 5.41 (t, J = 5.4 Hz) 1H), 4.50 (dd, J = 3.4, 11.7 Hz, 1H), 4.47 - 4.42 (m, 2H), 4.07 - 3.91 (m, 3H), 3.73 - 3.58 (m, 2H), 2.61 - 2.50 (m, 2H), 2.25 (dddd, J = 5.7, 7.1, 9.4, 14.2 Hz, 1H), 1.85 (d, J = 6.4 Hz, 3H). LCMS (APCI): 146.2 [M+H] + .

[0445] Step 5: Synthesis of ((3R,5S)-4-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidine-4-yl)-5-methyl-1,4-oxazepan-3-yl)methanol (I-12e) To a solution of ((3R,5S)-5-methyl-1,4-oxazepan-3-yl)methanol (430 mg, 1 equivalent, 2.96 mmol) in acetonitrile (50 mL) in an ice bath, Hünig base (1.53 g, 2.06 mL, 4 equivalents, 11.8 mmol) was added, followed by 4,5,7-trichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidine (I-1, 884 mg, 1 equivalent, 2.96 mmol). The reaction mixture was stirred in the ice bath and then warmed to room temperature. After 3 hours, the reaction was complete according to LC-MS. CH3CN was removed under vacuum, and the resulting bright orange solid was purified by flash column chromatography (Isco Gold, 80 g silica gel column, gradient of 0-5.5% MeOH in DCM, then held in 5.5% MeOH until the product eluted) to obtain ((3R,5S)-4-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidine-4-yl)-5-methyl-1,4-oxazepan-3-yl)methanol (1.23 g, 94%) as an orange solid. 1 ¹H NMR (400 MHz, methanol-d4) δ = 4.57 (dd, J = 5.5, 11.0 Hz, 1H), 4.45 (dd, J = 6.6, 11.0 Hz, 1H), 4.01 (dd, J = 3.8, 12.3 Hz, 1H), 3.88 - 3.65 (m, 4H), 3.54 - 3.46 (m, 1H), 3.19 - 3.08 (m, 1H), 2.66 (s, 3H), 1.93 - 1.83 (m, 1H), 1.76 - 1.64 (m, 1H), 1.20 (d, J = 6.4 Hz, 3H). 19 F NMR (377 MHz, chloroform (d) δ = -137.17 (s, 1F). LCMS (APCI): 407.0 [M+H] + .

[0446] Step 6: Synthesis of (8aS,13S)-5-chloro-4-fluoro-13-methyl-2-(methylthio)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene(I-12) To a turbid solution of ((3R,5S)-4-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidine-4-yl)-5-methyl-1,4-oxazepan-3-yl)methanol (1.07 g, 1 equivalent, 2.62 mmol) in acetonitrile (65 mL), DBU (1.198 g, 1.186 mL, 3.0 equivalents, 7.867 mmol) was added. The reaction mixture was stirred at 50°C for 20 hours. CH3CN was removed under vacuum to obtain a concentrated oily substance, which solidified upon standing. The solid was ground with CH3CN at 50°C. After stirring the slurry for 1 hour, it was removed from the oil bath and cooled to room temperature. The solid was filtered, washed with MTBE, and dried under vacuum to obtain (8aS,13S)-5-chloro-4-fluoro-13-methyl-2-(methylthio)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene (0.614 g, 63%) as an off-white powder. 1¹H NMR (400 MHz, chloroform-d) δ = 4.60 (dd, J = 5.1, 13.3 Hz, 1H), 4.38 (dd, J = 1.1, 13.3 Hz, 1H), 4.32 - 4.19 (m, 2H), 4.14 - 4.02 (m, 2H), 3.65 (dd, J = 10.4, 12.8 Hz, 1H), 3.28 (ddd, J = 4.6, 11.6, 12.8 Hz, 1H), 2.63 (s, 3H), 2.53 - 2.43 (m, 1H), 2.10 (dddd, J = 5.5, 6.5, 11.7, 14.0 Hz, 1H), 1.76 (d, J = 6.9 Hz, 3H). 19 F NMR (377 MHz, chloroform-d) δ = -140.42 (s, 1F). LCMS (APCI): 371.0 [M+H] + .

[0447] Intermediate 13(I-13):4-bromo-6-chloro-5-(1-methylcyclopropyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole

[0448] [ka]

[0449] Step 1: Synthesis of 4-chloro-2-fluoro-5-(propa-1-en-2-yl)aniline (I-13a) 5-bromo-4-chloro-2-fluoroaniline (2500 mg, 11.14 mmol), 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (2810 mg, 16.7 mmol), and K3PO4 (7090 mg, 33.4 mmol) were added to 1,4-dioxane (90 mL) and water (9 mL), to which Pd(dppf)Cl2 (815 mg, 1.11 mmol) was added. The mixture was stirred under a nitrogen atmosphere at 80 °C for 16 hours. LC-MS showed that the starting materials were consumed. The reaction product was diluted with water (30 mL) and extracted with ethyl acetate (3 × 20 mL). The organic layers were combined, washed with brine (15 mL), dried over Na2SO4, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by flash column chromatography (40 g silica gel, 0-5% gradient of ethyl acetate in PE) to obtain the title intermediate (1500 mg, 73%) as a yellow oily substance. 1 H NMR (400 MHz, CDCl3) δ 7.00 (d, J = 10.7 Hz, 1H), 6.62 (d, J = 9.4 Hz, 1H), 5.19 - 5.17 (m, 1H), 4.94 - 4.91 (m, 1H), 3.65 (br, 2H), 2.06 - 2.05 (m, 3H). LCMS (ESI) m / z: 186.1 [M+H]+.

[0450] Step 2: Synthesis of N-(4-chloro-2-fluoro-5-(propa-1-en-2-yl)phenyl)acetamide (I-13b) To a solution of 4-chloro-2-fluoro-5-(propa-1-en-2-yl)aniline (1500 mg, 8.08 mmol) in 1,4-dioxane (40 mL), acetic anhydride (1150 mg, 11.3 mmol) and DIPEA (1040 mg, 8.08 mmol) were added at 25 °C. The mixture was stirred at 50 °C for 20 hours. LC-MS showed that SM was consumed. The mixture was extracted with  (3 × 40 mL), the organic layers were combined, washed with brine (15 mL), dried over Na₂SO₄, filtered, and concentrated under vacuum to obtain N-(4-chloro-2-fluoro-5-(propa-1-en-2-yl)phenyl)acetamide (1700 mg, 92%) as a yellow solid. 1 H NMR (400 MHz, CDCl3) δ 8.20 (d, J = 8.5 Hz, 1H), 7.12 (d, J = 10.7 Hz, 1H), 5.26 - 5.21 (m, 1H), 4.97 (d, J = 0.7 Hz, 1H),2.22 (s, 3H), 2.08 - 2.05 (m, 3H). LCMS (ESI) m / z: 228 [M+H]+.

[0451] Step 3: Synthesis of N-(4-chloro-2-fluoro-5-(1-methylcyclopropyl)phenyl)acetamide (compound I-13c) A solution of diethylzinc (8300 mg, 67.2 mmol, 67.2 mL, 1.0 M) in DCE (40 mL) was mixed with a solution of TFA (7660 mg, 67.2 mmol, 5.15 mL) in DCE (15 mL) under a nitrogen atmosphere at 0°C. After 30 minutes, a solution of CH2I2 (18000 mg, 67.2 mmol, 5.41 mL) in DCE (10 mL) was added to the mixture and stirred for 30 minutes. Next, a solution of N-(4-chloro-2-fluoro-5-(propa-1-en-2-yl)phenyl)acetamide (1700 mg, 7.467 mmol) in DCE (15 mL) was added to the mixture. The reaction mixture was stirred at 25°C for 4 hours. LC-MS showed that the starting materials were almost completely consumed. The solvent was removed under reduced pressure. The crude product was purified by flash column chromatography (40 g silica gel, 0-10% ethyl ether in petroleum ether) to obtain N-(4-chloro-2-fluoro-5-(1-methylcyclopropyl)phenyl)acetamide (1500 mg, 83%) as a pale yellow solid. 1 H NMR (400 MHz, CDCl3) δ 8.28 (d, J = 8.6 Hz, 1H), 7.09 (d, J = 10.6 Hz, 1H), 2.24 (s, 3H), 1.32 (s, 3H), 0.83 - 0.79 (m, 2H),0.78 - 0.73 (m, 2H). LCMS (ESI) m / z: 242.1 [M+H]+.

[0452] Step 4: Synthesis of 4-chloro-2-fluoro-5-(1-methylcyclopropyl)aniline (I-13d) To a solution of N-(4-chloro-2-fluoro-5-(1-methylcyclopropyl)phenyl)acetamide (1500 mg, 6.206 mmol) in EtOH (50 mL), NaOH (9190 mg, 230 mmol) was added, and the mixture was stirred at 75°C for 16 hours. LC-MS showed that the starting material had been consumed. Most of the EtOH was removed under reduced pressure. The residue was diluted with ice water (50 mL) and adjusted to pH=5 with 2 M HCl. The mixture was extracted with siRNA (3 × 30 mL). The organic layers were combined, washed with brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by flash column chromatography (25 g silica gel, 0-10% siRNA in petroleum ether) to obtain 4-chloro-2-fluoro-5-(1-methylcyclopropyl)aniline (870 mg, 70%) as a pale yellow oil. 1 H NMR (400 MHz, CDCl3) δ 6.97 (d, J = 10.6 Hz, 1H), 6.77 (d, J = 9.5 Hz, 1H), 1.30 (s, 3H), 0.78 - 0.74 (m, 2H), 0.72 - 0.68 (m, 2H). LCMS (ESI) m / z: 200.1 [M+H]+.

[0453] Step 5: Synthesis of 2-bromo-4-chloro-6-fluoro-3-(1-methylcyclopropyl)aniline (I-13e) To a solution of 4-chloro-2-fluoro-5-(1-methylcyclopropyl)aniline (1040 mg, 5.209 mmol) in DMF (35 mL), NBS (927 mg, 5.21 mmol) was gradually added under a nitrogen atmosphere at 0°C. The mixture was stirred at 20°C for 16 hours. LC-MS showed that the starting material had been consumed. The mixture was diluted with saturated NaHCO3 aqueous solution (5 mL) and extracted with ethyl acetate (25 mL x 3). The organic layers were combined, washed with brine (10 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain 2-bromo-4-chloro-6-fluoro-3-(1-methylcyclopropyl)aniline (1320 mg, 91%) as a brown oil. 1H NMR (400 MHz, CDCl3) δ 7.02 (d, J = 10.4 Hz, 1H), 1.29 (s, 3H), 0.96 - 0.87 (m, 4H). LCMS (ESI) m / z: 278 [M+H]+.

[0454] Step 6: Synthesis of 3-bromo-1-chloro-5-fluoro-4-iodo-2-(1-methylcyclopropyl)benzene(I-13f) CuI (1130 mg, 5.92 mmol) and tert-butyl nitrite (611 mg, 5.92 mmol) were added to a flask containing CH3CN (20 mL). The mixture was stirred at 25°C for 30 minutes. 2-bromo-4-chloro-6-fluoro-3-(1-methylcyclopropyl)aniline (1100 mg, 3.95 mmol) was added dropwise to CH3CN (10 mL), and the mixture was stirred at 5°C for 20 minutes, then heated to 25°C for 40 hours. LC-MS showed that the starting material was almost completely consumed. The mixture was quenched with HCl (5 mL, 1 M) and extracted with RINKAN (3 × 20 mL). The combined organic layers were washed with saturated aqueous Na2SO3 (20 mL) and brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (4 g silica gel, petroleum ether) to obtain 3-bromo-1-chloro-5-fluoro-4-iodo-2-(1-methylcyclopropyl)benzene (1150 mg, 75%) as a pale yellow oil. 1 H NMR (400 MHz, CDCl3) δ 7.11 (d, J = 7.4 Hz, 1H), 1.31 (s, 3H), 0.98 - 0.85 (m, 4H).

[0455] Step 7: Synthesis of 2-bromo-4-chloro-6-fluoro-3-(1-methylcyclopropyl)benzaldehyde (I-13g) To a solution of 3-bromo-1-chloro-5-fluoro-4-iodo-2-(1-methylcyclopropyl)benzene (1050 mg, 2.696 mmol) in THF (40 mL), n-BuLi (225 mg, 3.51 mmol) was added dropwise at -78 °C under a nitrogen atmosphere. The mixture was stirred at that temperature for 10 minutes. Then, anhydrous DMF (394 mg, 5.39 mmol) was added to the mixture, and the mixture was stirred at -78 °C for 1 hour. LC-MS showed that the starting material was consumed and a new peak was formed. The mixture was quenched with 1N HCl (3 mL) and slowly warmed to 20 °C. The reaction mixture was then diluted with water and extracted with Â(3 × 15 mL). The organic layers were combined, washed with water, dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by flash column chromatography (12 g silica gel, PE) to obtain 2-bromo-4-chloro-6-fluoro-3-(1-methylcyclopropyl)benzaldehyde (440 mg, 56%) as a pale yellow oil. 1 H NMR (400 MHz, DMSO-d6) δ 10.18 (s, 1H), 7.70 (d, J = 10.4 Hz, 1H), 1.30 (s, 3H), 1.06 - 1.00 (m, 2H), 0.92 - 0.84 (m, 2H).

[0456] Step 8: Synthesis of 4-bromo-6-chloro-5-(1-methylcyclopropyl)-1H-indazole(I-13h) To a mixture of 1,4-dioxane (16 mL) and 2-bromo-4-chloro-6-fluoro-3-(1-methylcyclopropyl)benzaldehyde (480 mg, 1.65 mmol), 85% N2H4.H2O (291 mg, 4.94 mmol) was added at 0°C, and the mixture was then stirred at 95°C for 90 hours. LC-MS showed that the starting material had been consumed. The reaction mixture was cooled to 25°C and then slowly poured into water (10 mL). The mixture was extracted with MTBE (3 × 10 mL). The organic layer was dried over Na2SO4, filtered, and concentrated to obtain the crude product. The crude product was purified by flash column chromatography (silica gel 4 g, PE) to obtain 4-bromo-6-chloro-5-(1-methylcyclopropyl)-1H-indazole (300 mg, 58%) as a pale yellow oil. 1 H NMR (400 MHz, CDCl3) δ 8.06 (s, 1H), 7.51 (s, 1H), 1.37 (s, 3H), 1.02 - 0.90 (m, 4H). LCMS (ESI) m / z: 285.0 [M+H]+.

[0457] Step 9: Synthesis of 4-bromo-6-chloro-5-(1-methylcyclopropyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole (intermediate I-13) To a solution of 4-bromo-6-chloro-5-(1-methylcyclopropyl)-1H-indazole (270 mg 0.945 mmol) in DCM (9 mL), DHP (119 mg 1.42 mmol), followed by p-toluenesulfonic acid monohydrate (18 mg 0.0945 mmol), was added. The reaction mixture was stirred at 30°C for 2 hours. LC-MS showed that the starting materials were consumed. The reaction mixture was washed with saturated NaHCO3 aqueous solution (10 mL) and brine (10 mL). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (4 g silica gel, 0% to 5% PE with siRNA) to obtain 4-bromo-6-chloro-5-(1-methylcyclopropyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole (300 mg, 77%) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 7.98 (s, 1H), 7.63 - 7.60 (m, 1H), 5.63 - 5.61 (m, 1H), 4.03 - 3.95 (m, 1H), 3.73 (ddd, J = 15.2, 7.2, 3.9 Hz, 1H), 2.53 - 2.42 (m, 1H), 2.18 - 2.05 (m, 2H), 1.78 - 1.68 (m, 3H), 1.36 (s, 3H), 1.00 - 0.88 (m, 4H). LCMS (ESI) m / z: 369.0 [M+H]+.

[0458] Intermediate 14(I-14): Ethyl(S)-(2-cyclopropyridenetetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol

[0459] [ka]

[0460] Step 1: Synthesis of 1-(tert-butyl)-1,4-dihydro-5H-tetrazole-5-thion (I-14a) To a solution of 2-isothiocyanato-2-methylpropane (2300 mg, 19.97 mmol) in 2-propanol (30 mL), sodium azide (1630 mg, 24.0 mmol) in H2O (20 mL) was added at 25 °C. The reaction mixture was stirred under a nitrogen atmosphere at 110 °C for 16 hours. LC-MS showed that the desired product had been formed. The reaction mixture was quenched by dropwise addition of concentrated HCl and stirred at 25 °C for 1 hour. The mixture was neutralized by the addition of solid NaHCO3, extracted with siRNA (5 × 50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (12 g silica gel, 0-20% siRNA in petroleum ether) to obtain 1-(tert-butyl)-1,4-dihydro-5H-tetrazole-5-thion (850 mg, 27%) as a colorless oil. 1 H NMR (400 MHz, DMSO-d6) δ 5.62 (s, 1H), 1.76 (s, 9H). LCMS (ESI) m / z 159.1 (M+H) + .

[0461] Step 2: Synthesis of 5-((3-bromopropyl)thio)-1-(tert-butyl)-1H-tetrazole (I-14b) To a solution of 1-(tert-butyl)-1,4-dihydro-5H-tetrazole-5-thion (700 mg, 4.42 mmol) in THF (40 mL), DEAD (1160 mg, 6.64 mmol) and PPh3 (1740 mg, 6.64 mmol) were added. The mixture was cooled to 0°C, and 3-bromopropan-1-ol (738 mg, 6.64 mmol) was added. The resulting reaction mixture was stirred at 25°C for 3 hours. LCMS showed that the desired product had been formed. The reaction mixture was concentrated under reduced pressure and purified by flash column chromatography (12 g silica gel, 0-10% ethyl ether) to obtain 5-((3-bromopropyl)thio)-1-(tert-butyl)-1H-tetrazole (500 mg, 35%) as a colorless oil. 1H NMR (400 MHz, CDCl3) δ 3.59 - 3.49 (m, 4H), 2.49 - 2.36 (m, 2H), 1.73 (s, 9H). LCMS (ESI) m / z 279.0, 281.0 (M+H) + .

[0462] Step 3: Synthesis of 5-((3-bromopropyl)sulfonyl)-1-(tert-butyl)-1H-tetrazole(I-14c) A solution of 5-((3-bromopropyl)thio)-1-(tert-butyl)-1H-tetrazole (400 mg 1.43 mmol) in EtOH (10 mL) is mixed with Mo7O in H2O2 (30%, 5 mL). 24 A solution of (NH4)6.×H2O (334 mg, 0.287 mmol) was added at 0°C. The resulting reaction mixture was stirred at 25°C for 3 hours. LC-MS showed that the desired product had been formed. The reaction mixture was poured into a saturated aqueous solution of Na2SO3 (30 mL) and extracted with EA (30 mL × 3). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (12 g silica gel, 0-10% ethyl ether) to obtain 5-((3-bromopropyl)sulfonyl)-1-(tert-butyl)-1H-tetrazole (500 mg, 90%) as a colorless oil. 1 H NMR (400 MHz, CDCl3) δ 4.04 - 3.96 (m, 2H), 3.59 (t, J = 6.3 Hz, 2H), 2.63 - 2.53 (m, 2H), 1.86 (s, 9H). LCMS (ESI) m / z 311.0, 313.0 (M+H)+.

[0463] Step 4: Synthesis of 1-(tert-butyl)-5-(cyclopropylsulfonyl)-1H-tetrazole (I-14d) To a solution of 5-((3-bromopropyl)sulfonyl)-1-(tert-butyl)-1H-tetrazole (450 mg, 1.45 mmol) in THF (15 mL), NaHMDS (1.08 mL, 2.0 M in THF, 2.17 mmol) was added at -78°C. The resulting reaction mixture was stirred at -78°C for 1 hour. LC-MS showed that the desired product had formed. The reaction mixture was poured into a saturated aqueous solution of NH4Cl (30 mL) and extracted with EA (30 mL x 3). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (12 g silica gel, 0-2% MeOH in DCM) to obtain 1-(tert-butyl)-5-(cyclopropylsulfonyl)-1H-tetrazole (340 mg, 92%) as a colorless oil. 1 H NMR (400 MHz, CDCl3) δ 3.28 (tt, J = 7.9, 4.9 Hz, 1H), 1.85 (s, 9H), 1.53 - 1.48 (m, 2H), 1.42 - 1.35 (m, 2H). LCMS (ESI) m / z 231.1 (M+H) + .

[0464] Step 5: Synthesis of ethyl(S)-2-cyclopropyridene-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate(I-14e) To a solution of 1-(tert-butyl)-5-(cyclopropylsulfonyl)-1H-tetrazole (260 mg, 1.13 mmol) and ethyl(S)-2,5-dioxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (238 mg, 1.13 mmol) in THF (15 mL), KHMDS (1.69 mL, 1.0 M in THF, 1.69 mmol) was added at -78°C. The resulting reaction mixture was stirred at 25°C for 16 hours. LC-MS showed that the desired product had formed. The reaction mixture was poured into saturated NH4Cl aqueous solution (30 mL), extracted with EA (3 × 30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (12 g silica gel, 0-2% MeOH in DCM) to obtain (S)-2-cyclopropylidene-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (160 mg, 52%) as a colorless oil. 1 H NMR (400 MHz, CDCl3) δ 4.36 (d, J = 15.2 Hz, 1H), 4.27 - 4.13 (m, 3H), 3.78 (dd, J = 14.9, 7.7 Hz, 1H), 3.17 (d, J = 15.5 Hz, 1H), 3.04 - 2.94 (m, 1H), 2.86 - 2.75 (m, 1H), 2.67 - 2.41 (m, 4H), 2.25 - 2.10 (m, 2H), 1.33 - 1.26 (m, 3H). LCMS (ESI) m / z 236.1 (M+H) + .

[0465] Step 6: Synthesis of ethyl(S)-(2-cyclopropyridenetetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol (intermediate I-14) To a solution of (S)-2-cyclopropyridene-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (140 mg, 0.595 mmol) in THF (10 mL), LiAlH4 (90 mg, 2.38 mmol) was added at 25 °C. The resulting reaction mixture was stirred at 70 °C for 2 hours. LC-MS showed that the desired product had been formed. Na2SO4.10H2O (2 g) was added to the reaction mixture, stirred at 25 °C for 10 minutes, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (12 g silica gel, 0-10% MeOH in DCM) to obtain (S)-(2-cyclopropyridenetetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol (40 mg, 33%) as a colorless oil. 1 H NMR (400 MHz, CDCl3) δ 4.33 (d, J = 14.0 Hz, 1H), 3.94 - 3.79 (m, 3H), 3.59 (d, J = 13.8 Hz, 1H), 2.95 (d, J = 15.3 Hz, 1H), 2.91 - 2.79 (m, 1H), 2.56 (d, J = 15.2 Hz, 1H), 2.40 - 2.27 (m, 1H), 2.24 - 1.99 (m, 6H), 1.91 - 1.77 (m, 1H). LCMS (ESI) m / z 180.1 (M+H) + .

[0466] Intermediate 15(I-15): (S)-(2-(oxetane-3-ylidene)tetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol

[0467] [ka]

[0468] Step 1: Synthesis of 5-(oxetane-3-ylthio)-1-phenyl-1H-tetrazole (I-15a) To a solution of 1-phenyl-1,4-dihydro-5H-tetrazole-5-thion (500 mg, 2.81 mmol) in DMF (10 mL), K2CO3 (968 mg, 7.01 mmol) and 3-iodooxetane (1040 mg, 14.0 mmol) were added. The resulting reaction mixture was stirred at 90°C for 2 hours. The reaction mixture was poured into H2O (10 mL) and extracted with EA (2 × 10 mL). The combined organic layer was concentrated under reduced pressure and purified by flash column chromatography (24 g silica gel, 0-30% siRNA in petroleum ether) to obtain 5-(oxetane-3-ylthio)-1-phenyl-1H-tetrazole (710 mg, 98%) as a yellow solid. 1 H NMR (400 MHz, CDCl3) δ 7.66 - 7.51 (m, 5H), 5.22 (t, J = 7.3 Hz, 2H), 5.08 - 4.97 (m, 1H), 4.76 - 4.64 (m, 2H). LCMS (ESI) m / z: 235.2 [M+H] + .

[0469] Step 2: Synthesis of 5-(oxetane-3-ylsulfonyl)-1-phenyl-1H-tetrazole (I-15b) A solution of 5-(oxetane-3-ylthio)-1-phenyl-1H-tetrazole (650 mg, 2.77 mmol) in EtOH (36 mL) is added at 0°C to Mo7O in H2O2 (18 mL). 24 A solution of (NH4)6 (596 mg, 0.56 mmol) was added over 5 minutes. The reaction mixture was stirred at 20°C for 12 hours. The reaction mixture was quenched with an aqueous solution of Na2SO3 (10 mL) and then extracted with EA (20 mL x 3). The combined organic phase was washed with brine (3 x 20 mL), dried on sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (24 g silica gel, 0-20% ethyl ethyl ether in petroleum ether) to obtain 5-(oxetane-3-ylsulfonyl)-1-phenyl-1H-tetrazole (650 mg, 88%) as a white solid. 1H NMR (400 MHz, CDCl3) δ 7.74 (dd, J = 4.0, 2.0 Hz, 2H), 7.67 - 7.61 (m, 3H), 5.30 - 5.22 (m, 1H), 5.09 (d, J = 6.8 Hz, 4H). LCMS (ESI) m / z: 267.0 [M+H] + .

[0470] Step 3: Synthesis of ethyl(S)-2-(oxetane-3-ylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate(I-15c) A solution of 5-(oxetane-3-ylsulfonyl)-1-phenyl-1H-tetrazole (250 mg, 0.94 mmol) in THF (15 mL) was added to KHMDS (1.13 mL, 1.13 mmol, 1 M in THF) over 5 minutes at -78°C. The reaction mixture was stirred under a nitrogen atmosphere at -78°C for 50 minutes. Then, ethyl(S)-2,5-dioxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (198 mg, 0.94 mmol) in THF (5 mL) was added to the mixture and stirred at 0°C for 2 hours. The reaction mixture was quenched with saturated NH4Cl aqueous solution (30 mL) and then extracted with  (3 × 50 mL). The combined organic phase was washed with brine (3 × 20 mL), dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (12 g silica gel, 0-80% ethyl in petroleum ether) to obtain ethyl(S)-2-(oxetane-3-ylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (155 mg, 66%) as a colorless oil. 1 H NMR (400 MHz, CDCl3) δ 5.14 (s, 4H), 4.23 (q, J = 7.1 Hz, 2H), 4.15 (d, J = 15.3 Hz, 1H), 3.54 (d, J = 15.4 Hz, 1H), 2.88 (d, J = 15.7 Hz, 1H), 2.83 - 2.74 (m, 1H), 2.66 - 2.57 (m, 1H), 2.46 (dd, J = 16.7, 9.4 Hz, 1H), 2.27 (d, J = 15.9 Hz, 1H), 2.18 - 2.07 (m, 1H), 1.29 (t, J = 7.1 Hz, 3H). LCMS (ESI) m / z: 252.1 [M+H] + .

[0471] Step 4: Synthesis of (S)-(2-(oxetane-3-ylidene)tetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol(I-15) To a solution of ethyl(S)-2-(oxetane-3-ylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (110 mg, 0.44 mmol) in THF (8 mL), LiAlH4 (0.88 mL, 0.88 mmol, 1 M in THF) was added. The mixture was stirred under a nitrogen atmosphere at 70°C for 1 hour. The reaction product was then quenched with Na2SO4·10H2O and filtered. The filtrate was concentrated under reduced pressure to obtain (S)-(2-(oxetane-3-ylidene)tetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol (85 mg, 99%) as a white oily substance. 1 H NMR (400 MHz, MeOD) δ 5.18 - 5.14 (m, 4H), 3.58 - 3.55 (m, 2H), 3.48 - 3.40 (m, 1H), 3.18 - 3.10 (m, 1H), 3.05 - 2.98 (m, 1H), 2.67 - 2.60 (m, 1H), 2.49 - 2.37 (m, 1H), 2.20 - 2.08 (m, 1H), 1.98 - 1.87 (m, 2H), 1.78 - 1.67 (m, 2H). LCMS (ESI) m / z :196.1 [M+H] + .

[0472] Intermediate 16(I-16): (S)-(2-(tetrahydro-4H-pyran-4-ylidene)tetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol

[0473] [ka]

[0474] Step 1: Synthesis of 1-phenyl-5-((tetrahydro-2H-pyran-4-yl)thio)-1H-tetrazole (I-16a) To a solution of 1-phenyl-1,4-dihydro-5H-tetrazole-5-thion (500 mg, 2.81 mmol) in DMF (5 mL), K2CO3 (968 mg, 7.01 mmol) and 4-bromotetrahydro-2H-pyran (1160 mg, 7.01 mmol) were added. The resulting mixture was stirred at 90°C for 3 hours. LC-MS showed that the desired product had been formed. The reaction mixture was combined with a batch of 50 mg of starting material 1, poured into H2O (20 mL), and extracted with EA (2 × 10 mL). The combined organic layer was concentrated under reduced pressure and purified by flash column chromatography (24 g silica gel, 0-30% gradient of siRNA in petroleum ether) to obtain 1-phenyl-5-((tetrahydro-2H-pyran-4-yl)thio)-1H-tetrazole (630 mg, 78%) as a yellow solid. 1 H NMR (400 MHz, CDCl3) δ 7.63 - 7.49 (m, 5H), 4.25 - 4.16 (m, 1H), 3.99 (dt, J = 12.0, 3.8 Hz, 2H), 3.59 (td, J = 11.9, 2.3 Hz, 2H), 2.23 (dd, J = 11.6, 1.6 Hz, 2H), 1.91 - 1.77 (m, 2H). LCMS (ESI) m / z: 263.2 [M+H] + .

[0475] Step 2: Synthesis of 1-phenyl-5-((tetrahydro-2H-pyran-4-yl)sulfonyl)-1H-tetrazole (I-16b) A solution of 1-phenyl-5-((tetrahydro-2H-pyran-4-yl)thio)-1H-tetrazole (630 mg 2.4 mmol) in EtOH (16 mL) is mixed with Mo7O in H2O2 (30%, 8 mL). 24 A solution of (NH4)6 × H2O (559 mg, 0.480 mmol) was added at 0°C. The resulting mixture was stirred at 25°C for 3 hours. LC-MS showed that most of the starting material had been consumed and the desired product had been formed. The reaction mixture was poured into a saturated aqueous solution of Na2SO3 (30 mL) and extracted with ethyl acetate (3 × 30 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (12 g silica gel, 0-10% ethyl acetate in petroleum ether) to obtain 1-phenyl-5-((tetrahydro-2H-pyran-4-yl)sulfonyl)-1H-tetrazole (524 mg, 74%) as a white solid. 1 H NMR (400 MHz, CDCl3) δ 7.72 - 7.56 (m, 5H), 4.22 - 4.10 (m, 3H), 3.52 (t, J = 11.8 Hz, 2H), 2.21 - 2.13 (m, 2H), 2.05 (td, J = 12.1, 4.5 Hz, 2H). LCMS (ESI) m / z: 295.2 [M+H] + .

[0476] Step 3: Synthesis of ethyl(S)-5-oxo-2-(tetrahydro-4H-pyran-4-ylidene)tetrahydro-1H-pyrrolidine-7a(5H)-carboxylate(I16c) To a solution of 1-phenyl-5-((tetrahydro-2H-pyran-4-yl)sulfonyl)-1H-tetrazole (200 mg, 0.680 mmol) in THF (20 mL), LiHMDS (1 M in THF, 1.02 mL, 1.02 mmol) was added over 5 minutes at -78°C. The reaction mixture was stirred under a nitrogen atmosphere at -78°C for 50 minutes. Then, ethyl(S)-2,5-dioxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (172 mg, 0.815 mmol) in THF (5 mL) was added to the mixture and stirred at 0°C for 2 hours. LC-MS showed that most of the starting material was consumed and the desired product was formed. The reaction mixture was quenched with saturated NH4Cl aqueous solution (30 mL) and then extracted with EA (3 × 50 mL). The combined organic phases were washed with brine (3 × 20 mL), dried on sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (4 g silica gel, 0-100% ethyl in petroleum ether) to obtain ethyl(S)-5-oxo-2-(tetrahydro-4H-pyran-4-ylidene)tetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (52 mg, crude product) as a colorless oil. 1 H NMR (400 MHz, CDCl3) δ 4.32 (d, J = 15.3 Hz, 1H), 4.25 - 4.18 (m, 2H), 3.74 - 3.66 (m, 3H), 3.65 - 3.58 (m, 2H), 3.14 (d, J = 15.8 Hz, 1H), 2.83 - 2.75 (m, 1H), 2.66 - 2.58 (m, 1H), 2.50 - 2.42 (m, 2H), 2.35 (d, J = 16.0 Hz, 1H), 2.23 - 2.14 (m, 4H), 1.30 - 1.27 (m, 3H). LCMS (ESI) m / z: 280.3 [M+H] + .

[0477] Step 4: Synthesis of (S)-(2-(tetrahydro-4H-pyran-4-ylidene)tetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol (intermediate I-16) To a solution of ethyl(S)-5-oxo-2-(tetrahydro-4H-pyran-4-ylidene)tetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (52 mg, 0.19 mmol) in THF (5 mL), LiAlH4 (0.372 mL, 0.372 mmol) was added under a nitrogen atmosphere. The reaction mixture was then stirred at 70°C for 2 hours. LC-MS showed that the desired product was formed. The mixture was quenched with Na2SO4.10H2O and stirred at 25°C for 1 hour. The mixture was filtered and concentrated to obtain (S)-(2-(tetrahydro-4H-pyran-4-ylidene)tetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol (43 mg) as an oil, which was used for the next step without further purification. LC-MS (ESI) m / z: 224.3 [M+H] + .

[0478] Intermediate 17(I-17): (S,Z)-2-(7a-(hydroxymethyl)tetrahydro-1H-pyrrolidine-2(3H)-ylidene)-N,N-dimethylacetamide

[0479] [ka]

[0480] Step 1: Synthesis of (S)-7a-(hydroxymethyl)tetrahydro-1H-pyrrolidine-2(3H)-one (I-17a) (S)-(2-methylenetetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol (800 mg, 5.22 mmol) was dissolved in a mixture of TFA (22 mL) and DCM (22 mL, c=0.2 M) and cooled to -20°C. Ozone was bubbling into the mixture at -20°C for 20 minutes. -20°C was achieved with NaCl and ice water in a 1:3 ratio. LCMS showed that most of the starting material had been consumed and the desired product had been formed. The reaction was then quenched by bubbling nitrogen into the mixture for 30 minutes. The solution was concentrated under reduced pressure to obtain crude (S)-7a-(hydroxymethyl)tetrahydro-1H-pyrrolidine-2(3H)-one (2.4 g, crude) as a pale yellow oil. 1 H NMR (400 MHz, CDCl3) δ 4.31 (d, J = 17.8 Hz, 1H), 4.17 - 4.04 (m, 2H), 3.84 (d, J = 12.7 Hz, 1H), 3.52 (d, J = 17.9 Hz, 1H), 3.21 - 3.08 (m, 2H), 2.63 (d, J = 18.9 Hz, 1H), 2.34 - 2.23 (m, 3H), 2.18 - 2.10 (m, 1H). LCMS(ESI) m / z: 156.1 [M+H] + .

[0481] Step 2: Synthesis of (S)-7a-(((tert-butyldimethylsilyl)oxy)methyl)tetrahydro-1H-pyrrolidine-2(3H)-one (I-17b) To a solution of (S)-7a-(hydroxymethyl)tetrahydro-1H-pyrrolidine-2(3H)-one (2.4 g, 8.915 mmol) and DIPEA (6.9 g, 53.5 mmol) in DCM (40 mL), TBSCl (1.75 g, 11.6 mmol) in DCM (10 mL) was added dropwise at 0°C. The mixture was then stirred at 25°C for 2 hours. LC-MS showed that the desired product had been formed. The reaction mixture was diluted with DCM (50 mL), washed with water, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (40 g silica gel, 0-60% ethyl ether) to obtain (S)-7a-(((tert-butyldimethylsilyl)oxy)methyl)tetrahydro-1H-pyrrolidine-2(3H)-one (925 mg, 38.5%) as a pale yellow oil. 1 H NMR (400 MHz, CDCl3) δ 3.60 - 3.39 (m, 3H), 3.33 - 3.22 (m, 1H), 3.07 (d, J = 18.7 Hz, 1H), 2.69 - 2.57 (m, 1H), 2.47 (d, J = 18.2 Hz, 1H), 2.20 (dd, J = 18.2, 1.1 Hz, 1H), 1.85 - 1.83 (m, 4H), 0.83 (s, 9H), 0.02 - 0.00 (m, 6H). LCMS(ESI) m / z: 270.2 [M+H] +

[0482] Step 3: Synthesis of methyl(S)-2-(7a-(((tert-butyldimethylsilyl)oxy)methyl)tetrahydro-1H-pyrrolidine-2(3H)-ylidene)acetate(I-17c) To a solution of methyl 2-(dimethoxyphosphoryl)acetate (1050 mg, 5.79 mmol) in THF (20 mL), n-BuLi (1.74 mL, 4.34 mmol, 1.2 M in hexane) was added at 0°C. The mixture was stirred at 0°C for 10 minutes. Then, (S)-7a-(((tert-butyldimethylsilyl)oxy)methyl)tetrahydro-1H-pyrrolidine-2(3H)-one (780 mg, 2.89 mmol) in THF (5 mL) was added dropwise at 0°C. The resulting mixture was stirred at 20°C for 2 hours. LC-MS showed that the desired product had been formed. The reaction mixture was quenched with water (2 mL) and diluted with EA (80 mL). The solution was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (40 g silica gel, 0-60% butyl in petroleum ether) to obtain methyl(S)-2-(7a-(((tert-butyldimethylsilyl)oxy)methyl)tetrahydro-1H-pyrrolidine-2(3H)ylidene) acetate (560 mg, 59%) as a pale yellow oil. 1 ¹H NMR (400 MHz, CDCl3) δ 5.82 and 5.74 (2s, 1H, subisomer and main isomer), 3.71 and 3.69 (2s, 3H, main isomer and subisomer), 3.48 - 3.39 (m, 2H), 3.33 - 3.03 (m, 2H), 2.88 - 2.52 (m, 2H), 1.96 - 1.82 (m, 3H), 1.78 - 1.57 (m, 3H), 0.87 and 0.86 (2s, 9H, subisomer and main isomer), 0.03 and 0.02 (2s, 6H, major isomer and subisomer). LC-MS (ESI) m / z: 326.1 [M+H] +

[0483] Step 4: Synthesis of (S)-2-(7a-(((tert-butyldimethylsilyl)oxy)methyl)tetrahydro-1H-pyrrolidine-2(3H)-ylidene)acetic acid (I-17d) A solution of methyl(S)-2-(7a-(((tert-butyldimethylsilyl)oxy)methyl)tetrahydro-1H-pyrrolidine-2(3H)-ylidene) acetate (480 mg, 1.47 mmol) and LiOH (106 mg, 4.42 mmol) in THF / MeOH / H2O (6 mL / 6 mL / 6 mL) was stirred at 25°C for 16 hours. LC-MS showed that the desired product was formed. The reaction mixture was diluted with water (30 mL) and the pH was adjusted to 6 with 0.3 M HCl. The solution was lyophilized to obtain crude (S)-2-(7a-(((tert-butyldimethylsilyl)oxy)methyl)tetrahydro-1H-pyrrolidine-2(3H)-ylidene)acetic acid (480 mg, crude) as a white solid. LC-MS (ESI) m / z: 312.1 [M+H] +

[0484] Step 5: Synthesis of (S)-2-(7a-(((tert-butyldimethylsilyl)oxy)methyl)tetrahydro-1H-pyrrolidine-2(3H)-ylidene)-N,N-dimethylacetamide(I-17e) (S)-2-(7a-(((tert-butyldimethylsilyl)oxy)methyl)tetrahydro-1H-pyrrolidine-2(3H)-ylidene)acetic acid (480 mg, 1.54 mmol) and DIPEA (996 mg, 7.70 mmol) in DCM (30 mL) were mixed with HATU (703 mg, 1.85 mmol) and dimethylamine hydrochloride (251 mg, 3.08 mmol). The mixture was then stirred at 25°C for 2 hours. LC-MS showed that the desired product had formed. The reaction mixture was diluted with DCM (20 mL) and washed with water (40 mL x 3). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (12 g silica gel, 0-50% siRNA in petroleum ether for 15 minutes, held for 5 minutes, then 0-20% MeOH in DCM for 15 minutes) to obtain (S)-2-(7a-(((tert-butyldimethylsilyl)oxy)methyl)tetrahydro-1H-pyrrolidine-2(3H)-ylidene)-N,N-dimethylacetamide (197 mg, 38%) as a yellow oil.1 H NMR (400 MHz, CDCl3) δ 5.48 (s, 1H), 3.73 - 3.70 (m, 4H), 3.30 (d, J = 16.5 Hz, 1H), 3.11 - 3.01 (m, 5H), 2.99 - 2.95 (m, 4H), 2.09 - 1.99 (m, 3H), 1.86 (s, 1H), 0.89 (d, J = 2.9 Hz, 9H), 0.10 (d, J = 4.0 Hz, 6H). LCMS(ESI) m / z: 339.2 [M+H] +

[0485] Step 6: Chiral separation of (S,Z)-2-(7a-(((tert-butyldimethylsilyl)oxy)methyl)tetrahydro-1H-pyrrolidine-2(3H)-ylidene)-N,N-dimethylacetamide(I-17f) (S)-2-(7a-(((tert-butyldimethylsilyl)oxy)methyl)tetrahydro-1H-pyrrolidine-2(3H)-ylidene)-N,N-dimethylacetamide (197 mg, 0.270 mmol) was subjected to chiral separation (SFC150; column: Daicel CHIRALCEL IC, 250 mm × 30 mm). The mixture was purified using an ID of 10 μM; mobile phase: CO2 / MeOH [0.2% NH3 (7M solution in MeOH)] = 70 / 30; flow rate: 780 g / min; wavelength: UV 214 nm; temperature: 35 °C) to obtain (S,Z)-2-(7a-(((tert-butyldimethylsilyl)oxy)methyl)tetrahydro-1H-pyrrolidine-2(3H)-ylidene)-N,N-dimethylacetamide (95 mg, 48%, Rt: 2.997 min, peak 2) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 5.50 (s, 1H), 3.81 (d, J = 15.2 Hz, 1H), 3.47 - 3.39 (m, 2H), 3.34 (d, J = 15.2 Hz, 1H), 3.19 (d, J = 15.8 Hz, 1H), 3.16 - 3.10 (m, 1H), 3.07 (d, J = 15.3 Hz, 1H), 2.98 (s, 3H), 2.93 (s, 3H), 2.56 (dt, J = 9.7, 7.1 Hz, 1H), 1.91 - 1.83 (m, 1H), 1.78 - 1.69 (m, 2H), 1.64 - 1.53 (m, 1H), 0.86 (d, J = 2.7 Hz, 9H), 0.00 (d, J = 2.8 Hz, 6H). LCMS(ESI) m / z: 339.3 [M+H] +

[0486] Step 7: Synthesis of (S,Z)-2-(7a-(hydroxymethyl)tetrahydro-1H-pyrrolidine-2(3H)-ylidene)-N,N-dimethylacetamide (intermediate I-17) To a solution of (S,Z)-2-(7a-(((tert-butyldimethylsilyl)oxy)methyl)tetrahydro-1H-pyrrolidine-2(3H)-ylidene)-N,N-dimethylacetamide (70 mg, 0.21 mmol) in DCM (5 mL), HCl / dioxane (4.0 M, 2 mL) was added dropwise, and the mixture was stirred at 25°C for 30 minutes. LC-MS showed that the desired product was formed. The mixture was diluted with DCM (5 mL), washed with water (10 mL), and subsequently extracted with water (10 mL × 2). The aqueous phase was freeze-dried to obtain (S,Z)-2-(7a-(hydroxymethyl)tetrahydro-1H-pyrrolidine-2(3H)-ylidene)-N,N-dimethylacetamide (55 mg, crude) as a yellow oil. 1 H NMR (400 MHz, MeOD) δ 5.55 (s, 1H), 4.39 (d, J = 14.4 Hz, 1H), 3.95 (d, J = 15.3 Hz, 1H), 3.75 (dd, J = 29.2, 12.1 Hz, 2H), 3.68 - 3.56 (m, 1H), 3.45 - 3.34 (m, 2H), 3.29 - 3.25 (m, 1H), 3.07 (s, 3H), 2.95 (d, J = 3.8 Hz, 3H), 2.23 - 1.97 (m, 4H). LCMS (ESI) m / z 225.1 [M+H] + .

[0487] Intermediate 18(I-18): (S,Z)-(2-(2-((tert-butyldimethylsilyl)oxy)ethylidene)tetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol

[0488] [ka]

[0489] Step 1: Synthesis of ethyl(S)-2-(2-acetoxyethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate(I-18a) A solution of ethyl(S)-2-methylene-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (450 mg, 2.15 mmol) and (Z)-buta-2-ene-1,4-diyldiacetate (741 mg, 4.30 mmol) in DCM (13 mL) was mixed with a second-generation Grubbs catalyst (183 mg, 0.215 mmol). The mixture was then stirred under a nitrogen atmosphere at 40 °C for 16 hours. LC-MS showed that the starting materials were consumed and the desired product was formed. The solution was concentrated under reduced pressure. The residue was purified by flash column chromatography (12 g silica gel, 0-80% ethyl in petroleum ether) to obtain ethyl(S)-2-(2-acetoxyethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (373 mg, 62%) as a black oil. 1 H NMR (400 MHz, CDCl3) δ 5.58 (ddd, J = 6.8, 5.6, 2.6 Hz, 1H), 4.57 - 4.45 (m, 2H), 4.42 - 4.31 (m, 1H), 4.25 - 4.08 (m, 2H), 3.83 (d, J = 16.1 Hz, 0.7H, major isomer), 3.74 (d, J = 12.8 Hz, 0.3H, minor isomer), 3.27 (d, J = 16.4 Hz, 0.3H, minor isomer), 3.07 (d, J = 16.0 Hz, 0.7H, main isomer), 2.85 - 2.73 (m, 1H), 2.70 - 2.57 (m, 1H), 2.56 - 2.37 (m, 2H), 2.19 - 2.09 (m, 1H), 2.06 - 2.04 (m, 3H), 1.27 (t, J = 7.1 Hz, 3H). LCMS (ESI) m / z: 282.1

[0490] Step 2: Synthesis of ethyl(S)-2-(2-hydroxyethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate(I-18b) To a solution of ethyl(S)-2-(2-acetoxyethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (350 mg, 1.24 mmol) in EtOH (10 mL), K2CO3 (344 mg, 2.49 mmol) was added. The reaction mixture was then stirred at 25°C for 1 hour. LC-MS showed that the starting material was consumed and the desired product was formed. The reaction mixture was then filtered to remove the base, and the filtrate was diluted with H2O (50 mL) and extracted with ELISA (50 mL x 3). The organic layers were then combined and concentrated under reduced pressure to obtain crude ethyl(S)-2-(2-hydroxyethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (185 mg, crude) as a black oil, which was used for the next step without further purification. 1 H NMR (400 MHz, CDCl3) δ 5.70 - 5.55 (m, 1H), 4.33 (d, J = 16.2 Hz, 1H), 4.21 (q, J = 7.1 Hz, 2H), 4.17 - 4.06 (m, 2H), 3.85 - 3.69 (m, 1H), 3.23 (d, J = 16.0 Hz, 0.3H, minor isomer), 3.07 (d, J = 15.8 Hz, 0.7H, main isomer), 2.85 - 2.73 (m, 1H), 2.67 - 2.57 (m, 1H), 2.55 - 2.41 (m, 2H), 2.18 - 2.07 (m, 1H), 1.28 (t, J = 7.1 Hz, 3H). LCMS (ESI) m / z: 240.7 [M+H] + .

[0491] Step 3: Synthesis of ethyl(S)-2-(2-((tert-butyldimethylsilyl)oxy)ethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate(I-18c) To a solution of ethyl(S)-2-(2-hydroxyethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (100 mg, 0.418 mmol) in DCM (5 mL), TBSCl (126 mg, 0.836 mmol), TEA (84.6 mg, 0.836 mmol), and DMAP (21.4 mg, 0.176 mmol) were added. The reaction mixture was then stirred under a nitrogen atmosphere at 25°C for 48 hours. LC-MS showed that the starting materials were consumed and the desired product was formed. The reaction mixture was diluted with H2O (20 mL) and extracted with DCM (20 mL x 3). The combined organic layers were washed with brine (100 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (4.0 g silica gel, 0% to 100% DCM in PE) to obtain ethyl(S)-2-(2-((tert-butyldimethylsilyl)oxy)ethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (141 mg, 95%) as a yellow oil. 1H NMR (400 MHz, CDCl3) δ 5.52 - 5.44 (m, 1H), 4.22 (d, J = 16.7 Hz, 1H), 4.18 - 4.04 (m, 4H), 3.73 - 3.63 (m, 1H), 3.12 (d, J = 16.0 Hz, 0.3H, minor isomer), 2.98 (d, J = 15.6 Hz, 0.7H, main isomer), 2.72 (dt, J = 16.6, 10.0 Hz, 1H), 2.60 - 2.50 (m, 1H), 2.46 - 2.34 (m, 2H), 2.11 - 2.03 (m, 1H), 1.21 (t, J = 7.1 Hz, 3H), 0.83 (s, 9H), 0.00 (s, 6H). LCMS (ESI) m / z: 354.3 [M+H] + .

[0492] Step 4: Synthesis of ethyl(S,Z)-2-(2-((tert-butyldimethylsilyl)oxy)ethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate(I-18d) Ethyl(S)-2-(2-((tert-butyldimethylsilyl)oxy)ethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (120 mg) was purified by chiral separation (instrument: SFC 150; column: Daicel CHIRALCEL AS, 250 mm × 30 mm ID, 10 μm; mobile phase: CO2 / MeOH [0.2% NH3 (7 M solution in MeOH)] = 90 / 10; flow rate: 80 g / min; wavelength: UV 214 nm; temperature: 35 °C) to obtain ethyl(S,Z)-2-(2-((tert-butyldimethylsilyl)oxy)ethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (50 mg, peak 1 of 2, Rt: 1.584 min, yield: 43%). 1H NMR (400 MHz, CDCl3) δ 5.52 - 5.43 (m, 1H), 4.21 (d, J = 15.9 Hz, 1H), 4.13 (q, J = 7.1 Hz, 2H), 4.06 (dd, J = 3.8, 1.2 Hz, 2H), 3.69 (d, J = 15.8 Hz, 1H), 2.98 (d, J = 15.6 Hz, 1H), 2.71 (dt, J = 16.7, 10.0 Hz, 1H), 2.53 (ddd, J = 13.1, 9.1, 1.7 Hz, 1H), 2.47 - 2.33 (m, 2H), 2.12 - 2.01 (m, 1H), 0.83 (s, 9H), -0.00 (s, 6H).

[0493] Step 5: Synthesis of (S,Z)-(2-(2-((tert-butyldimethylsilyl)oxy)ethylidene)tetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol(I-18) To a solution of ethyl(S,Z)-2-(2-((tert-butyldimethylsilyl)oxy)ethylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (50 mg, 0.14 mmol) in THF (5 mL), LAH (0.283 mL, 1 M in THF) was added at 0°C under a nitrogen atmosphere. The resulting mixture was heated to 70°C and stirred at 70°C for 1 hour. LC-MS showed that the starting material was consumed and the desired product was detected. The mixture was quenched with Na2SO4.10H2O and stirred at 25°C for 2 hours. The mixture was filtered and concentrated under reduced pressure to obtain the product (S,Z)-(2-(2-((tert-butyldimethylsilyl)oxy)ethylidene)tetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol (60 mg, crude) as a pale yellow oil, which was used for the next step without further purification. LCMS(ESI) m / z: 298.3 [M+H] + This was carried out according to the steps of general scheme A, with a final overall deprotection step using HCl in dioxane at 25°C.

[0494] Intermediate 19(I-19):(1R,7aS * )-7a-(hydroxymethyl)-6-methylenehexahydro-1H-pyrrolidine-1-ol

[0495] [ka]

[0496] Step 1: Synthesis of 2-(((tert-butyldimethylsilyl)oxy)methyl)propa-2-en-1-ol (I-19a) Sodium hydride (4.99 g, 125 mmol) was added at 0°C to a mixture of 2-methylene-1,3-propanediol (10 g, 113.5 mmol) in THF (400 mL). The mixture was stirred under a nitrogen atmosphere at 0°C for 1 hour. Tert-butyldimethylsilyl chloride (18.0 g, 119 mmol) was added in one batch. The mixture was stirred at 25°C for 16 hours. LCMS was performed to obtain the desired product (UV214, R) after the starting material was consumed. t The formation of (1.360 min) was indicated. Water (200 mL) was added. The mixture was extracted with siRNA (400 mL x 3). The combined organic layer was dried over Na2SO4, filtered, and concentrated to dryness. The crude product was purified by flash column chromatography (CH2Cl2 in 0%~30% PE) to obtain 2-(((tert-butyldimethylsilyl)oxy)methyl)propa-2-en-1-ol (23.2 g, crude product) as a colorless oil. 1 H NMR (400 MHz, CDCl3) δ 5.11 - 5.09 (m, 1H), 5.08 (dd, J = 2.4, 1.2 Hz, 1H), 4.25 (s, 2H), 4.17 (s, 2H), 0.91 (d, J = 2.6 Hz, 9H), 0.10 - 0.08 (m, 6H). LCMS(m / z) =203.2 (M+H) +

[0497] Step 2: Synthesis of tert-butyl((2-(iodomethyl)allyl)oxy)dimethylsilane(I-19b) Imidazole (5.37 g, 78.9 mmol) and PPh3 (20.7 g, 78.9 mmol) were added at 0°C to a mixture of 2-(((tert-butyldimethylsilyl)oxy)methyl)propa-2-en-1-ol (13.3 g, 65.72 mmol) in CH2Cl2 (300 mL). I2 (20 g, 78.9 mmol) was added all at once. The reaction mixture was stirred at 0°C, and then stirred at 25°C for 3 hours. TLC (PE=100%, I 2、 A Rf of 0.8 indicated that the starting material had been consumed and new spots had been detected. The mixture was concentrated under reduced pressure. The residue was diluted with CH2Cl2 (50 mL). Then, hexane (300 mL) was added. The mixture was filtered through Celite. The filtrate was concentrated to dryness. The residue was purified by flash column chromatography (220 g silica gel, PE=100%) to obtain tert-butyl((2-(iodomethyl)allyl)oxy)dimethylsilane (15.3 g, 75%) as a pale yellow oil. 1 H NMR (400 MHz, CDCl3) δ 5.31 (d, J = 0.8 Hz, 1H), 5.19 (dd, J = 2.8, 1.4 Hz, 1H), 4.31 (s, 2H), 3.95 (s, 2H), 0.92 (s, 9H), 0.10 (d, J = 3.1 Hz, 6H).

[0498] Step 3: Synthesis of 1-(tert-butyl)2-methyl(2R,3R)-3-hydroxypyrrolidine-1,2-dicarboxylate(I-19c) To a solution of (2R,3R)-1-(tert-butoxycarbonyl)-3-hydroxypyrrolidine-2-carboxylic acid (2 g, 8.65 mmol) in DMF (50 mL), K2CO3 (3.6 g, 25.9 mmol) and CH3I (3.68 g, 25.9 mmol) were added at 0°C. The reaction mixture was stirred at 25°C for 16 hours. LC-MS showed that the starting material was consumed and the desired product was detected. The reaction mixture was diluted with saturated LiCl aqueous solution (100 mL) and extracted with  (50 mL x 3). The combined organic layers were washed with water (100 mL) and brine (100 mL) and dried over Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (elution with phenyl in 0%-35% PE on 24 g of silica gel) to obtain 1-(tert-butyl)2-methyl(2R,3R)-3-hydroxypyrrolidine-1,2-dicarboxylate (2.3 g, crude product) as a pale yellow oil. 1 H NMR (400 MHz, CDCl3) δ 4.44 (s, 1H), 4.31 - 4.22 (m, 1H), 3.74 (s, 3H), 3.69 - 3.52 (m, 2H), 2.16 - 2.04 (m, 1H), 1.92 - 1.90 (m, 1H), 1.44 (m, 9H). LCMS (ESI) m / z 268.1 (M+Na) + .

[0499] Step 4: Synthesis of 1-(tert-butyl)2-methyl(2R,3R)-3-methoxypyrrolidine-1,2-dicarboxylate (I-19d) A mixture of 1-(tert-butyl)2-methyl(2R,3R)-3-hydroxypyrrolidine-1,2-dicarboxylate (4.4 g, 17.9 mmol), CH3I (25.2 g, 179 mmol), and Ag2O (24.9 g, 108 mmol) in CH3CN (75 mL) was stirred at 20°C for 20 hours. LC-MS showed that some of the starting material remained and the desired product was detected. Ag2O (6.24 g, 26.9 mmol) was added to the above solution and stirred for a further 24 hours. LC-MS showed that most of the starting material was consumed and the desired product was detected. The reaction mixture was filtered, and the filter cake was washed with CH3CN (30 mL). The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (elution with ethyl in 0%-25% PE on 24 g of silica gel) to obtain 1-(tert-butyl)2-methyl(2R,3R)-3-methoxypyrrolidine-1,2-dicarboxylate (3.95 g, 85%) as a colorless oil. 1 H NMR (400 MHz, CDCl3) δ 4.42 - 4.26 (m, 1H), 3.96 - 3.84 (m, 1H), 3.75 (d, J = 2.8 Hz, 3H), 3.70 3.42 (m, 2H), 3.38 (s, 3H), 2.10 - 1.94 (m, 2H), 1.45 - 1.43 (m, 9H). LCMS (ESI) m / z 282.0 (M+Na).

[0500] Step 5: Synthesis of 1-(tert-butyl)2-methyl(3R)-2-(2-(((tert-butyldimethylsilyl)oxy)methyl)allyl)-3-methoxypyrrolidine-1,2-dicarboxylate(I-19e) To a mixture of 1-(tert-butyl)2-methyl(2S,3S)-3-methoxypyrrolidine-1,2-dicarboxylate (3.9 g, 15.04 mmol) in THF (65 mL), LiHMDS (30.1 mL, 30.1 mmol, 1 M in THF) was added at -65°C. The mixture was stirred at -65°C for 1 hour. Tert-butyl((2-(iodomethyl)allyl)oxy)dimethylsilane (9.39 g, 30.1 mmol) in THF (5 mL) was added dropwise to the above solution at -65°C. The mixture was warmed to 20°C and stirred under a nitrogen atmosphere at 20°C for 16 hours. LCMS showed that the starting material was consumed and the desired product was formed. The reaction mixture was quenched with saturated NH4Cl aqueous solution (60 mL) and then extracted with ELISA (50 mL x 3). The combined organic phases were washed with brine (60 mL), dried on sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (elution with ethyl in 40 g silica gel and 0%-5% PE) to obtain 1-(tert-butyl)2-methyl(3R)-2-(2-(((tert-butyldimethylsilyl)oxy)methyl)allyl)-3-methoxypyrrolidine-1,2-dicarboxylate (2.2 g, 33%) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 5.39 - 5.25 (m, 1H), 4.97 (d, J = 12.2 Hz, 1H), 4.17 - 3.97 (m, 3H), 3.75 - 3.68 (m, 3H), 3.51 - 3.28 (m, 4H), 3.26 - 3.08 (m, 1H), 3.02 - 2.77 (m, 1H), 2.61 - 2.54 (m, 1H), 2.13 - 1.87 (m, 2H), 1.45 - 1.41 (m, 9H), 0.91 (s, 9H), 0.05 (s, 6H). LCMS(ESI) m / z = 466.1 (M+Na) +

[0501] Step 6: Synthesis of methyl(3R)-2-(2-(chloromethyl)allyl)-3-methoxypyrrolidine-2-carboxylate(I-19f) SOCl2 (4.2 mL) was slowly added to a solution of 1-(tert-butyl)2-methyl(3R)-2-(2-(((tert-butyldimethylsilyl)oxy)methyl)allyl)-3-methoxypyrrolidine-1,2-dicarboxylate (0.3 g, 0.67 mmol) in CH2Cl2 (10 mL). The reaction mixture was stirred at 20°C for 16 hours. LC-MS indicated that the reaction was complete. The mixture was concentrated under reduced pressure to obtain methyl(3R)-2-(2-(chloromethyl)allyl)-3-methoxypyrrolidine-2-carboxylate (230 mg, crude) as a brown oil, which was used directly for the next step. LC-MS (ESI) m / z 248.1 (M+H) + .

[0502] Step 7: Methyl(1R,7aS) * Synthesis of )-1-methoxy-6-methylenetetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (I-19g and I-19h) Crude methyl(3R)-2-(2-(chloromethyl)allyl)-3-methoxypyrrolidine-2-carboxylate (1600 mg, 6.45 mmol), sodium bicarbonate (2.71 g, 32.2 mmol), and potassium iodide (107 mg, 0.64 mmol) in CH3CN (70 mL) were added to a 25 mL round-bottom flask. The mixture was stirred at 20 °C for 1.5 hours. LC-MS showed that the starting material was consumed and the desired product was detected. The mixture was concentrated to remove most of the CAN, then diluted with H2O (50 mL) and extracted with i-PrOH / CH2Cl2 (1 / 5, 50 mL × 3). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (elution with siRNA in 24 g silica gel, 0%-40% PE (0.1% NH4OH)) to obtain methyl(1R,7aS) *)-1-methoxy-6-methylenetetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (intermediate I-19g) (150mg, 15%) was obtained as a yellow oily substance. 1 H NMR (400 MHz, MeOD) δ 5.06 - 4.91 (m, 2H), 4.13 - 4.07 (m, 1H), 3.92 - 3.86 (m, 1H), 3.75 (s, 3H), 3.46 - 3.41 (m, 1H), 3.35 (s, J = 3.7 Hz, 3H), 3.24 - 3.18 (m, 1H), 3.16 - 3.09 (m, 1H), 2.88 - 2.78 (m, 1H), 2.75 - 2.68 (m, 1H), 2.11 - 2.01 (m, 2H).[α] D 25 = +2.31(c 0.13, MeOH). And methyl(1R,7aR) * )-1-methoxy-6-methylenetetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (intermediate I-19h) (370 mg, 21%) was obtained as a yellow oily substance. 1 H NMR (400 MHz, MeOD) δ 5.06 - 4.93 (m, 2H), 3.97 - 3.91 (m, 1H), 3.73 (s, 3H), 3.70 - 3.65 (m, 1H), 3.41 - 3.34 (m, 2H), 3.30 (s, 3H), 3.09 - 3.00 (m, 1H), 2.78 - 2.68 (m, 1H), 2.62 - 2.56 (m, 1H), 2.24 - 2.14 (m, 1H), 2.05 - 1.96 (m, 1H). LCMS(ESI) m / z = 212 (M+1).α] D 25 = -24.62 (c 0.13, MeOH).

[0503] Step 8: ((1R, 7aS *Synthesis of )-1-methoxy-6-methylenetetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol(I-19j) Methyl(1R,7aS) in THF (15mL) * To a solution of 19g of 1-1-methoxy-6-methylenetetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (intermediate I) (150 mg, 0.71 mmol), DIBALH (4.73 mL, 1.5 M in toluene) was added at 0°C under a nitrogen atmosphere. The resulting mixture was heated to 25°C and stirred for 45 minutes at 25°C. LC-MS showed that the starting material was consumed and the desired product was detected. The mixture was quenched with MeOH (1 mL) at -50°C, then heated to 20°C and stirred for 30 minutes at 20°C. The mixture was then poured into an aqueous solution of sodium potassium tartrate tetrahydrate (10 mL, 2 M) at 20°C and stirred for 2 hours at 20°C. The organic layer was separated, and the aqueous layer was extracted with i-PrOH / CH2Cl2 (1 / 5, 15 mL × 4). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain ((1R,7aS * )-1-methoxy-6-methylenetetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol (compound 8-1) (130 mg, crude product) was obtained as an oily substance and used for the next step without further purification. LCMS (ESI) m / z 184.1 (M+H) + .

[0504] (1R,7aS * Synthesis of )-7a-(hydroxymethyl)-6-methylenehexahydro-1H-pyrrolidine-1-ol (intermediate I-19) CH2Cl2 (5 mL) contains ((1R, 7aS *To a solution of )-1-methoxy-6-methylenetetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol (compound 10-1) (130 mg, 0.71 mmol), BBr3 (1.78 g, 7.09 mmol) was added at 0°C. The mixture was heated to 20°C and stirred at 20°C for 16 hours. LC-MS showed that the starting material had been consumed and a major peak with the desired MS was detected. The mixture was concentrated under reduced pressure. The residue was diluted with CH2Cl2 (10 mL), quenched with MeOH (1 mL), and then basicized with NaHCO3 at 0°C. The separated aqueous layer was concentrated and then diluted with MeOH / CH2Cl2 (50 mL, 1 / 20). The mixture was filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography (elution with MeOH in 4g silica gel, 15%-100% CH2Cl2 (0.1% NH4OH)) to obtain the crude product. The crude product was diluted with MeOH / CH2Cl2 (20 mL, 1 / 10), then filtered, and concentrated under reduced pressure. The residue was diluted with CH3CN (50 mL), then ultrasonically washed, filtered, and concentrated under reduced pressure to obtain (1R, 7aR). * )-7a-(hydroxymethyl)-6-methylenehexahydro-1H-pyrrolidine-1-ol (compound 11-1) (75 mg, crude product) was obtained as a yellow oily substance. LCMS(ESI) m / z: 170.1 [M+H] + .

[0505] Intermediate 20(I-20): 6-Chloro-5-cyclopropyl-1-(oxan-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole

[0506] [ka] To a solution of 4-bromo-6-chloro-5-cyclopropyl-1-(oxan-2-yl)-1H-indazole (I-5ii, 30.0 g, 81.8 mmol) in tert-amyl alcohol (401 mL), bis(pinacolato)diborane (41.6 g, 163 mmol), palladium acetate (1.15 g, 5.11 mmol), triphenylphosphine (2.68 g, 10.2 mmol), and K3PO4 (52.1 g, 245 mmol) were added. The resulting mixture was degassed three times with Ar and then heated in an oil bath at 90°C for 16 hours. Analysis by LC-MS showed that the desired product had been formed. The mixture was filtered through Celite and rinsed with EtOac. The crude product was purified by flash column chromatography (ISCO, 120 g silica gel, acetone in 0-10% heptane) to obtain the title intermediate, I-20 (26.3 g, 80%). 1 H NMR (400 MHz, CDCl3) δ 8.14 (d, J = 1.0 Hz, 1H), 7.67 (d, J = 1.0 Hz, 1H), 5.64 (dd, J = 9.0, 2.9 Hz, 1H), 3.98 (dtd, J = 11.5, 3.7, 1.4 Hz, 1H), 3.77 - 3.60 (m, 1H), 2.57 - 2.39 (m, 1H), 2.13 (tt, J = 8.4, 5.6 Hz, 2H), 2.07 - 1.97 (m, 1H), 1.83 - 1.59 (m, 3H), 1.45 (s, 12H), 1.11 - 1.01 (m, 2H), 0.61 - 0.50 (m, 2H). MS: 403.2, [M+H] + . (Note: This procedure outlines a general method for converting indazole bromide to boronate, and can be applied with minor modifications and / or substitutions to obtain the desired boronate from the indazole bromide described therein.)

[0507] Intermediate 21(I-21): (8aS)-5-chloro-4-fluoro-2-(methanesulfonyl)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene

[0508] [ka] (S)-2-chloro-1-fluoro-12-(methylthio)-5a,6,9,10-tetrahydro-5H,8H-4,7-dioxa-3,10a,11,13-tetraazanaphtho[1,8-ab]heptalene (I-2, 3.0 g, 8.4 mmol, 1 equivalent) was dissolved in methyl ethyl ketone (84 mL) and saturated NaHCO3 aqueous solution (42 mL), and then oxone (11.6 g, 18.9 mmol) was added at room temperature. An additional oxone (14.1 g) was added gradually over 2 hours to achieve complete conversion, at which point the layers were separated, the aqueous layer was extracted with SiO2 (3 × 5 mL), the combined organic layers were dried over sodium sulfate, filtered, and concentrated. The residue was azeotropically mixed with CH3CN (2 × 5 mL) and concentrated again to obtain the desired product, which was collected as a crude product without purification.

[0509] Synthesis of intermediate 22(I-22):4-bromo-1-(tetrahydro-2H-pyran-2-yl)-5,6,7,8-tetrahydro-1H-benzo[f]indazole

[0510] [ka]

[0511] Step 1: Synthesis of 5-bromo-7-fluoro-1,2,3,4-tetrahydronaphthalene (I-22a) To a stirred solution of 8-bromo-6-fluoro-3,4-dihydronaphthalene-1(2H)-one (500 mg, 2.06 mmol) in TFA (2 mL), Et3SiH (1200 mg, 10.3 mmol) was added dropwise under nitrogen at 0°C. The mixture was stirred at 20°C for 16 hours. LC-MS showed that the starting material had been consumed. The mixture was concentrated under reduced pressure, quenched with saturated NH4Cl aqueous solution (10 mL), and extracted with siRNA (10 mL x 2). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (12 g silica gel, 0-2% siRNA in petroleum ether) to obtain 5-bromo-7-fluoro-1,2,3,4-tetrahydronaphthalene (460 mg, 97.6%) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 7.13 (dd, J = 8.1, 2.6 Hz, 1H), 6.76 (dd, J = 9.1, 2.5 Hz, 1H), 2.75 (t, J = 6.2 Hz, 2H), 2.68 (t, J = 6.2 Hz, 2H), 1.87 - 1.78 (m, 2H), 1.77 - 1.67 (m, 2H).

[0512] Step 2: Synthesis of 1-bromo-3-fluoro-5,6,7,8-tetrahydronaphthalene-2-carboaldehyde (I-22b) In an oven-dried vial equipped with a magnetic stirring rod under a nitrogen atmosphere, 2,2,6,6-tetramethylpiperidine (425 mg, 3.01 mmol) dissolved in THF (8 mL) was added, and the solution was cooled to -65°C. Then, n-BuLi (0.80 mL, 2.0 mmol) was added to the above solution. The reaction mixture was stirred at -40°C for 30 minutes. A solution of 5-bromo-7-fluoro-1,2,3,4-tetrahydronaphthalene (230 mg, 1.0 mmol) in THF (5 mL) was added to the above solution at -60°C, and stirring was continued at -60°C for 1 hour. Then, DMF (147 mg, 2.0 mmol) was added to the above solution at -60°C. The reaction mixture was stirred at -60°C for 1 hour. LC-MS showed that the starting material had been consumed and a new peak was detected. The mixture was quenched with saturated NH4Cl aqueous solution (2 mL) and extracted with siRNA (10 mL x 2). The organic layer was washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (12 g silica gel, 0-25% siRNA in petroleum ether) to obtain 1-bromo-3-fluoro-5,6,7,8-tetrahydronaphthalene-2-carboaldehyde (230 mg, 89.1%) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 10.38 (s, 1H), 6.88 (d, J = 11.0 Hz, 1H), 2.82 (t, J = 6.2 Hz, 2H), 2.76 (t, J = 6.2 Hz, 2H), 1.91 - 1.81 (m, 2H), 1.80 - 1.69 (m, 2H).

[0513] Step 3: Synthesis of 4-bromo-5,6,7,8-tetrahydro-1H-benzo[f]indazole (I-22c) To a stirred solution of 1-bromo-3-fluoro-5,6,7,8-tetrahydronaphthalene-2-carboaldehyde (230 mg, 0.895 mmol) in 1,4-dioxane (5 mL), N2H4.H2O (158 mg, 2.68 mmol) was added, and the mixture was stirred in a sealed tube at 120°C for 16 hours. LC-MS showed that the starting material was consumed and the desired product was detected. The mixture was quenched with saturated NH4Cl aqueous solution (2 mL) and extracted with siRNA (10 mL x 2). The organic layer was washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (12 g silica gel, 0-25% siRNA in petroleum ether) to obtain 4-bromo-5,6,7,8-tetrahydro-1H-benzo[f]indazole (95 mg, 42%) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 8.03 (s, 1H), 7.18 (s, 1H), 2.94 (t, J = 6.2 Hz, 2H), 2.89 (t, J = 6.6 Hz, 2H), 1.90 - 1.84 (m, 2H), 1.83 - 1.77 (m, 2H). LCMS (ESI) m / z 251, 253 [M+H] + .

[0514] Synthesis of 4-bromo-1-(tetrahydro-2H-pyran-2-yl)-5,6,7,8-tetrahydro-1H-benzo[f]indazole (I-22) To a stirred solution of 4-bromo-5,6,7,8-tetrahydro-1H-benzo[f]indazole (95 mg, 0.38 mmol) in DCM (2 mL), p-toluenesulfonic acid (7.2 mg, 0.038 mmol) and DHP (95 mg, 1.13 mmol) were added at 0°C under a nitrogen atmosphere. The mixture was stirred at 20°C for 1 hour. LC-MS showed that the starting material was consumed and the desired product was detected. The reaction mixture was poured into  (5 mL), washed with water (5 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (12 g silica gel, 0-30%  in petroleum ether) to obtain 4-bromo-1-(tetrahydro-2H-pyran-2-yl)-5,6,7,8-tetrahydro-1H-benzo[f]indazole (95 mg, yield: 75%) as a yellow oil. 1 HNMR (400 MHz, CDCl3) δ 8.10 (s, 1H), 7.40 (s, 1H), 5.70 (dd, J = 9.4, 2.8 Hz, 1H), 3.86 (dd, J = 10.7, 8.0 Hz, 1H), 3.55 - 3.46 (m, 1H), 2.91 - 2.87 (m, 4H), 1.92 - 1.80 (m, 4H), 1.80 - 1.70 (m, 6H). LCMS (ESI) m / z 335, 337 [M+H] + .

[0515] Synthesis of intermediates 23(I-23i and I-23ii):rel-4-bromo-6-chloro-5-((1R,2R)-2-methylcyclopropyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole

[0516] [ka]

[0517] Step 1: Synthesis of (E)-4-chloro-2-fluoro-5-(propa-1-en-1-yl)aniline (I-23a) Pd(dppf)Cl2 (1110 mg, 1.51 mmol) was added to a mixture of 5-bromo-4-chloro-2-fluoroaniline (3400 mg, 15.1 mmol), (E)-propa-1-en-1-ylboronic acid (2600 mg, 30.27 mmol), and K3PO4 (9640 mg, 45.4 mmol) in 1,4-dioxane (70 mL) and H2O (14 mL). The mixture was stirred under a nitrogen atmosphere at 110 °C for 16 hours. LC-MS detected the desired product and indicated that no starting materials remained. The reaction mixture was filtered. The filtrate was diluted with ELISA (200 mL) and washed with brine (100 mL x 2). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (80 g silica gel, 0-10% ethyl ethanol in petroleum ether) to obtain (E)-4-chloro-2-fluoro-5-(propa-1-en-1-yl)aniline (3140 mg, yield 97%) as a yellow oil. 1 H NMR (400 MHz, DMSO) δ 7.11 (d, J = 11.0 Hz, 1H), 6.97 (d, J = 9.6 Hz, 1H), 6.53 (dd, J = 15.7, 1.7 Hz, 1H), 6.09 - 6.05 (m, 1H), 5.41 - 5.07 (br, 2H), 1.86 (dd, J = 6.7, 1.6 Hz, 3H). LCMS (ESI) m / z: 186.0 [M+H] + .

[0518] Step 2: Synthesis of (E)-N-(4-chloro-2-fluoro-5-(propa-1-en-1-yl)phenyl)acetamide (I-23b) To a solution of (E)-4-chloro-2-fluoro-5-(propa-1-en-1-yl)aniline (2600 mg, 14.01 mmol) in dioxane (75 mL), acetic anhydride (2140 mg, 21.0 mmol) and DIPEA (2720 mg, 21.0 mmol) were added at 25 °C. The mixture was stirred at 60 °C for 16 hours. LC-MS showed that the starting material was consumed and the desired product was detected. The reaction mixture was diluted with ethyl acetate (150 mL) and washed with brine (150 mL x 2). The organic matter was dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was dissolved in ethyl acetate. Petroleum ether was then added dropwise until no more solids were formed. The mixture was filtered to obtain a white solid as (E)-N-(4-chloro-2-fluoro-5-(propa-1-en-1-yl)phenyl)acetamide (2200 mg, 58%). 1 H NMR (400 MHz, DMSO-d6) δ 9.81 (s, 1H), 8.14 (d, J = 8.4 Hz, 1H), 7.44 (d, J = 10.5 Hz, 1H), 6.61 (dd, J = 15.7, 1.7 Hz, 1H), 6.30 - 6.10 (m, 1H), 2.09 (s, 3H), 1.88 (dd, J = 6.6, 1.6 Hz, 3H). LCMS(ESI) (m / z): 228.1 [M+H] + .

[0519] Step 3: Synthesis of rac-N-(4-chloro-2-fluoro-5-((1R,2R)-2-methylcyclopropyl)phenyl)acetamide(I-23c) A solution of ZnEt2 (67.2 mmol, 67.2 mL, 1.0 M) in DCE (15 mL) was added to a solution of TFA (67.2 mmol, 5.15 mL) in DCE (20 mL) under a nitrogen atmosphere at 0°C. After 30 minutes, a solution of CH2I2 (67.2 mmol, 5.41 mL) in DCE (15 mL) was added to the mixture. After stirring for 30 minutes, a solution of (E)-N-(4-chloro-2-fluoro-5-(propa-1-en-1-yl)phenyl)acetamide (1700 mg, 7.467 mmol) in DCE (57 mL) was added to the mixture. The reaction mixture was stirred at 0°C for 2 hours. LC-MS showed that the starting material was consumed and the desired product was detected. The mixture was concentrated under reduced pressure. The crude product was purified by flash column chromatography (80 g silica gel, 0-10% phenylethylamine in petroleum ether) to obtain rac-N-(4-chloro-2-fluoro-5-((1R,2R)-2-methylcyclopropyl)phenyl)acetamide (1800 mg, 100%) as a pale yellow oily substance. 1 H NMR (400 MHz, CDCl3) δ 7.78 (d, J = 8.3 Hz, 1H), 7.12 (d, J = 10.4 Hz, 1H), 2.29 (s, 3H), 1.80 - 1.74 (m, 1H), 1.22 (d, J =5.7 Hz, 3H), 0.99 - 0.93 (m, 2H), 0.77 - 0.72 (m, 1H). LCMS(ESI) (m / z): 241.9 [M+H] + .

[0520] Step 4: Synthesis of rac-4-chloro-2-fluoro-5-((1R,2R)-2-methylcyclopropyl)aniline (I-23d) To a solution of rac-N-(4-chloro-2-fluoro-5-((1R,2R)-2-methylcyclopropyl)phenyl)acetamide (3200 mg, 13.24 mmol) in EtOH (130 mL), NaOH (21200 mg, 530 mmol) was added, and the mixture was stirred at 75°C for 16 hours. LC-MS showed that the starting material was consumed and the desired product was detected. Most of the EtOH was evaporated under reduced pressure. The residue was then diluted with ice water (50 mL) and adjusted to pH=5 with 2 M HCl. The mixture was extracted with siRNA (40 mL x 3). The organic layers were combined, washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by flash column chromatography (40 g silica gel, 0-10% ethyl ether in petroleum ether) to obtain rac-4-chloro-2-fluoro-5-((1R,2R)-2-methylcyclopropyl)aniline (1100 mg, 42%) as a pale yellow oil. 1 H NMR (400 MHz, CDCl3) δ 6.99 (d, J = 10.6 Hz, 1H), 6.34 (d, J = 9.3 Hz, 1H), 1.76 - 1.70 (m, 1H), 1.21 (d, J = 5.8 Hz, 3H),0.92 - 0.85 (m, 1H), 0.83 - 0.78 (m, 1H), 0.72 - 0.66 (m, 1H). LCMS (ESI) m / z: 200.2 [M +H] + .

[0521] Step 5: Synthesis of rac-2-bromo-4-chloro-6-fluoro-3-((1R,2R)-2-methylcyclopropyl)aniline (I-23e) To a solution of rac-4-chloro-2-fluoro-5-((1R,2R)-2-methylcyclopropyl)aniline (1100 mg, 5.51 mmol) in DMF (27 mL), NBS (981 mg, 5.51 mmol) was added under a nitrogen atmosphere at 0°C. The mixture was stirred at 20°C for 16 hours. LC-MS showed that the starting material was consumed and the desired product was detected. The mixture was diluted with saturated aqueous NaHCO3 (15 mL) and extracted with ethyl acetate (25 mL x 3). The combined organic layers were washed with brine (15 mL), dried over Na2SO4, filtered, and concentrated to obtain rac-2-bromo-4-chloro-6-fluoro-3-((1R,2R)-2-methylcyclopropyl)aniline (1400 mg, 91%) as a brown oil. 1 H NMR (400 MHz, CDCl3) δ 7.02 (d, J = 10.4 Hz, 1H), 1.38 - 1.32 (m, 1H), 1.28 (d, J = 5.9 Hz, 3H), 1.06 - 0.99 (m, 1H), 0.96 - 0.85 (m, 2H). LCMS (ESI) m / z: 278.0, 280.0 [M+H] + .

[0522] Step 6: Synthesis of rac-3-bromo-1-chloro-5-fluoro-4-iodo-2-((1R,2R)-2-methylcyclopropyl)benzene (I-23f) To a solution of CH3CN (20 mL), CuI (1440 mg, 7.54 mmol) and tert-butyl nitrite (778 mg, 7.54 mmol) were added. The mixture was stirred at 25°C for 30 minutes. rac-2-bromo-4-chloro-6-fluoro-3-((1R,2R)-2-methylcyclopropyl)aniline (1400 mg, 5.026 mmol) from CAN (20 mL) was added dropwise, and the mixture was stirred at 5°C for 20 minutes, then heated to 25°C for 40 hours. LC-MS showed that the starting material had been consumed and a new peak had formed. The mixture was quenched with HCl (10 mL, 1 M) and extracted with ethyl acetate (3 × 25 mL). The combined organic layers were washed with saturated aqueous Na2SO3 (30 mL) and brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (4 g silica gel, petroleum ether) to obtain rac-3-bromo-1-chloro-5-fluoro-4-iodo-2-((1R,2R)-2-methylcyclopropyl)benzene (1300 mg, 66%) as a pale yellow oil. 1 H NMR (400 MHz, CDCl3) δ 7.11 (d, J = 7.4 Hz, 1H), 1.46 - 1.40 (m, 1H), 1.29 (d, J = 5.6 Hz, 3H), 1.07 - 0.98 (m, 2H), 0.90 - 0.84 (m, 1H).

[0523] Step 7: Synthesis of rac-2-bromo-4-chloro-6-fluoro-3-((1R,2R)-2-methylcyclopropyl)benzaldehyde (I-23g) To a solution of rac-3-bromo-1-chloro-5-fluoro-4-iodo-2-((1R,2R)-2-methylcyclopropyl)benzene (1500 mg 3.852 mmol) in THF (40 mL), n-BuLi (321 mg 5.01 mmol) was added dropwise under nitrogen at -78 °C and stirred for 10 minutes. Then, anhydrous DMF (563 mg 7.7 mmol) was added to the above mixture and the mixture was stirred at -78 °C for 2 hours. LCMS showed that the starting material was consumed and a new peak was formed. The mixture was quenched with 1N HCl (10 mL) and slowly warmed to 20 °C. Then the mixture was diluted with water and extracted with EA (20 × 3 mL). The combined organic layers were washed with H2O, dried over Na2SO4, filtered, and concentrated to obtain the crude product. The crude product was purified by flash column chromatography (12 g silica gel, petroleum ether) to obtain rac-2-bromo-4-chloro-6-fluoro-3-((1R,2R)-2-methylcyclopropyl)benzaldehyde (510 mg, 45%) as a pale yellow oil. 1 H NMR (400 MHz, CDCl3) δ 10.33 (s, 1H), 7.20 (d, J = 10.0 Hz, 1H), 1.41 - 1.39 (m, 1H), 1.32 (d, J = 4.8 Hz, 3H), 1.10 - 1.01 (m, 2H), 0.95 - 0.87 (m, 1H).

[0524] Step 8: Synthesis of rac-4-bromo-6-chloro-5-((1R,2R)-2-methylcyclopropyl)-1H-indazole(I-23h) A mixture of rac-2-bromo-4-chloro-6-fluoro-3-((1R,2R)-2-methylcyclopropyl)benzaldehyde (510 mg, 1.75 mmol) in 1,4-dioxane (17 mL) was to be mixed with 85% N2H4.H2O (309 mg, 5.25 mmol) at 20 °C, and the mixture was then stirred at 95 °C for 24 hours. LC-MS showed that the starting material was consumed and the desired product was detected. The reaction mixture was cooled to 25 °C and slowly added to water (10 mL). The mixture was extracted with MTBE (30 mL). The organic layer was dried over Na2SO4, filtered, and concentrated to obtain the crude product. The crude product was purified by flash column chromatography (4 g silica gel, PE) to obtain rac-4-bromo-6-chloro-5-((1R,2R)-2-methylcyclopropyl)-1H-indazole (270 mg, 54%) as a pale yellow solid. LCMS (ESI) m / z: 285.0, 287.0 [M+H] + .

[0525] Step 9: Synthesis of rel-4-bromo-6-chloro-5-((1R,2R)-2-methylcyclopropyl)-1H-indazole (I-23i-1 and I-23i-2) rac-4-bromo-6-chloro-5-((1R,2R)-2-methylcyclopropyl)-1H-indazole (350 mg, 1.23 mmol) was purified by chiral separation (instrument: SFC 150; column: CHIRALPAK IK, 0.46 cm ID × 25 cm L, 0.5 μl; mobile phase: A / B: hexane / IPA = 95 / 5; flow rate: 1 mL / min; wavelength: UV 214 nm; temperature: 35°C) to obtain rel-4-bromo-6-chloro-5-((1R,2R)-2-methylcyclopropyl)-1H-indazole. Peak 1 was a colorless oily substance (135 mg, 38.6%), with [α]25D = +288.00 (c = 0.05, MeOH). LCMS (ESI) m / z: 285.0, 287.0 [M+H] + . 1 1H NMR (400 MHz, CDCl3) δ 8.06 (s, 1H), 7.52 (d, J = 0.8 Hz, 1H), 1.49 (dt, J = 8.4, 5.4 Hz, 1H), 1.33 (d, J = 5.9 Hz, 3H), 1.14 - Peak 1.06 (m, 1H), 1.03 - 0.94 (m, 2H). Peak 2 was a colorless oily substance (142 mg, 40.6%), [α]25D = -226.00 (c = 0.05, MeOH). LCMS (ESI) m / z: 285.0, 287.0 [M+H] + . 1 1H NMR (400 MHz, CDCl3) δ 8.06 (s, 1H), 7.52 (d, J = 0.7 Hz, 1H), 1.49 (dt, J = 8.4, 5.4 Hz, 1H), 1.33 (d, J = 5.9 Hz, 3H), 1.14 - 1.06 (m, 1H), 1.03 - 0.93 (m, 2H).

[0526] Step 10: Synthesis of rel-4-bromo-6-chloro-5-((1R,2R)-2-methylcyclopropyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole (I-23i and I-23ii) To a solution of rel-4-bromo-6-chloro-5-((1R,2R)-2-methylcyclopropyl)-1H-indazole (100 mg 0.35 mmol, I-23i-1) in DCM (4 mL), DHP (44.2 mg 0.525 mmol), followed by p-toluenesulfonic acid monohydrate (6.66 mg 0.0350 mmol), the reaction mixture was stirred at 25°C for 2 hours. LC-MS showed that the starting material was consumed and the desired product was detected. The reaction mixture was washed with saturated NaHCO3 aqueous solution (10 mL) and brine (10 mL). The organic layer was dried over Na2SO4, filtered, and concentrated to obtain rel-4-bromo-6-chloro-5-((1R * ,2R *)-2-methylcyclopropyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole (I-23i, 120 mg, Y: 93%) was obtained as a yellow oily substance. 1 H NMR (400 MHz, CDCl3) δ 7.97 (s, 1H), 7.62 (d, J = 0.6 Hz, 1H), 5.62 (dd, J = 9.1, 2.8 Hz, 1H), 4.04 - 3.95 (m, 1H), 3.78 - 3.69 (m, 1H), 2.53 - 2.42 (m, 1H), 2.18 - 2.10 (m, 1H), 2.09 - 2.02 (m, 1H), 1.81 - 1.68 (m, 4H), 1.32 (d, J = 5.9 Hz, 3H), 1.11 - 1.04 (m, 1H), 1.00 - 0.92 (m, 2H). LCMS (ESI) m / z: 369 [M+H] + .

[0527] Similarly, treat I-23i-2 as described above to obtain rel-4-bromo-6-chloro-5-((1R * ,2R * )-2-methylcyclopropyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole (I-23ii, 90 mg, 70%) was obtained as a colorless oil. 1 H NMR (400 MHz, CDCl3) δ 7.97 (s, 1H), 7.61 (d, J = 0.7 Hz, 1H), 5.62 (dd, J = 9.1, 2.8 Hz, 1H), 4.04 - 3.95 (m, 1H), 3.77 - 3.68 (m, 1H), 2.53 - 2.41 (m, 1H), 2.17 - 2.02 (m, 2H), 1.76 - 1.65 (m, 3H), 1.42 (d, J = 3.1 Hz, 1H), 1.33 (d, J = 6.1 Hz, 3H), 1.13 - 1.03 (m, 1H), 1.00 - 0.92 (m, 2H). LCMS (ESI) m / z: 369 [M+H] + .

[0528] Intermediate 24 (I-24i and I-24ii): (R * Synthesis of )-4-bromo-5-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-5,6,7,8-tetrahydro-1H-benzo[f]indazole

[0529] [ka]

[0530] Step 1: Synthesis of 6-fluoro-3,4-dihydronaphthalene-1(2H)-one O-methyloxime (I-24a) A mixture of 6-fluoro-1-tetralone (90 g, 548.19 mmol), NH2OMe·HCl (68.7 g, 822 mmol), and pyridine (65 g, 822 mmol) in CH3CN (500 mL) was stirred at 25°C for 2 hours. TLC (PE, UV) showed that the starting materials were consumed and the desired spots were detected. H2O (1000 mL) was added to the reaction mixture and extracted with MTBE (500 mL x 2). The combined organic layer was then washed with H2O (500 mL) and brine, dried over Na2SO4, filtered, and concentrated under vacuum. The brown foam was ground with toluene (40 mL) to obtain 6-fluoro-3,4-dihydronaphthalene-1(2H)-one O-methyloxime (98 g, 92.5%) as a light brown oil. 1 H NMR (400 MHz, CDCl3) δ 7.96 (dd, J = 8.8, 6.0 Hz, 1H), 6.88 (td, J = 8.6, 2.7 Hz, 1H), 6.82 (dd, J = 9.2, 2.6 Hz, 1H), 3.97 (s, 3H), 2.77 - 2.65 (m, 4H), 1.88 - 1.79 (m, 2H). LCMS (ESI) m / z: 194.1 [M+H] + .

[0531] Step 2: Synthesis of 8-bromo-6-fluoro-3,4-dihydronaphthalene-1(2H)-one O-methyloxime (I-24b) Palladium acetate (8.44 g, 50.7 mmol) was added under N2 to a solution of 6-fluoro-3,4-dihydronaphthalene-1(2H)-one O-methyloxime (98 g, 510 mmol) and NBS (108 g, 609 mmol) in acetic acid (800 mL). The reaction mixture was then stirred at 30°C for 12 hours. LC-MS showed that most of the starting material was consumed and a major peak with the desired mass was detected. The mixture was diluted with water (1000 mL) and TBME (800 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated under vacuum. The brown foam was ground with CH3CN (100 mL) to obtain the crude product 8-bromo-6-fluoro-3,4-dihydronaphthalene-1(2H)-one O-methyloxime (130 g, 94%) as a brown oil, which was used as is for the next step. 1 H NMR (400 MHz, CDCl3) δ 7.30 - 7.26 (m, 1H), 6.84 (dd, J = 8.3, 2.6 Hz, 1H), 4.03 (s, 3H), 2.75 (t, J = 6.9 Hz, 2H), 2.65 - 2.58 (m, 2H), 1.79 - 1.71 (m, 2H). LCMS (ESI) m / z: 272.0 [M+H] +

[0532] Step 3: Synthesis of 8-bromo-6-fluoro-3,4-dihydronaphthalene-1(2H)-one (I-24c) Crude 8-bromo-6-fluoro-3,4-dihydronaphthalene-1(2H)-one O-methyloxime (130 g, 478 mmol) in 1,4-dioxane (2000 mL) was mixed with H2SO4 (2460 mL, 4 M) at 25 °C. The mixture was stirred at 110 °C for 3 hours. LC-MS showed that the starting material was consumed and the desired product was detected. The reaction mixture was made basic to pH=8 with aqueous NaOH (1 M, 10 L). The mixture was extracted with ELISA (1000 mL x 2). The organic layers were combined, washed with brine (1000 mL), dried over Na2SO4, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by column chromatography (SiO2, 0-3% siRNA in petroleum ether) to obtain 8-bromo-6-fluoro-3,4-dihydronaphthalene-1(2H)-one (66 g, 56.8%) as a yellow solid. 1 H NMR (400 MHz, CDCl3) δ 7.31 (dd, J = 8.3, 2.5 Hz, 1H), 6.94 (dd, J = 8.4, 2.5 Hz, 1H), 2.98 (t, J = 6.2 Hz, 2H), 2.73 - 2.66 (m, 2H), 2.10 (dt, J = 12.8, 6.5 Hz, 2H). LCMS (ESI) m / z: 243 [M+H] + .

[0533] Step 4: Synthesis of 8-bromo-6-fluoro-1,2,3,4-tetrahydronaphthalene-1-ol (I-24d) To a solution of 8-bromo-6-fluoro-3,4-dihydronaphthalene-1(2H)-one (1 g, 4.114 mmol) in MeOH (20 mL), NaBH4 (311 mg, 8.23 ​​mmol) was added at 0°C. The mixture was stirred at 25°C for 12 hours. LC-MS showed that the starting material was consumed and the desired product was detected. The mixture was quenched with water (10 mL). The residue was extracted with DCM (20 mL x 2). The combined organic phase was dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by flash column chromatography (25 g silica gel column, 0-10% siRNA in petroleum ether) to obtain 8-bromo-6-fluoro-1,2,3,4-tetrahydronaphthalene-1-ol (846 mg, 83.9%) as a white solid. 1 H NMR (400 MHz, CDCl3) δ 7.18 (dd, J = 8.0, 2.5 Hz, 1H), 6.82 (dd, J = 9.0, 2.5 Hz, 1H), 4.99 (t, J = 3.2 Hz, 1H), 2.84 (dd, J = 17.2, 5.2 Hz, 1H), 2.76 - 2.65 (m, 1H), 2.28 (s, 1H), 2.24 - 2.16 (m, 1H), 2.04 - 1.91 (m, 1H), 1.81 - 1.69 (m, 2H). LCMS (ESI) m / z: 227 [M-17] + .

[0534] Step 5: Synthesis of 8-bromo-6-fluoro-1-methoxy-1,2,3,4-tetrahydronaphthalene (I-24e) 8-bromo-6-fluoro-1,2,3,4-tetrahydronaphthalene-1-ol (745 mg, 3.04 mmol) was added at 0°C to a solution of NaH (122 mg, 3.04 mmol) in THF (15 mL). The mixture was stirred at 0°C for 1 hour. Then, CH3I (647 mg, 4.56 mmol) was added at 0°C. The mixture was stirred at 25°C for 12 hours. LC-MS showed that the starting material was consumed and the desired product was detected. The mixture was quenched with aqueous NH4Cl (5 mL) and concentrated under reduced pressure. The residue was extracted with siRNA (10 mL x 2). The combined organic phase was dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by flash column chromatography (12 g silica gel column, 0-10% ethyl ether in petroleum ether) to obtain 8-bromo-6-fluoro-1-methoxy-1,2,3,4-tetrahydronaphthalene (700 mg, 89%) as a yellow oily substance. 1 H NMR (400 MHz, DMSO) δ 7.41 (dd, J = 8.4, 2.6 Hz, 1H), 7.04 (dd, J = 9.5, 2.6 Hz, 1H), 4.29 (t, J = 2.8 Hz, 1H), 3.36 (s, 3H), 2.84 - 2.76 (m, 1H), 2.65 (ddd, J = 17.5, 11.1, 6.6 Hz, 1H), 2.30 - 2.21 (m, 1H), 1.77 - 1.63 (m, 2H), 1.52 - 1.42 (m, 1H). LCMS (ESI) m / z :227[M-31] + .

[0535] Step 6: Synthesis of 1-bromo-3-fluoro-8-methoxy-5,6,7,8-tetrahydronaphthalene-2-carboaldehyde (I-24f) In an oven-dried vial equipped with a magnetic stirring rod under a nitrogen atmosphere, 2,2,6,6-tetramethylpiperidine (3590 mg, 25.5 mmol) dissolved in THF (10 mL) was added, and the solution was cooled to -65°C. Then, n-BuLi (6.79 mL, 17 mmol) was added to the above solution. The reaction mixture was stirred at -40°C for 30 minutes. Next, a solution of 8-bromo-6-fluoro-1-methoxy-1,2,3,4-tetrahydronaphthalene (2200 mg, 8.49 mmol) in THF (40 mL) was added to the above solution. Stirring was then continued at -60°C for 1 hour. Then, DMF (1240 mg, 17 mmol) was added to the above solution at -60°C. The resulting mixture was stirred at -60°C for 1 hour. LC-MS showed that the starting material was consumed and a small peak with the desired mass was detected. The mixture was quenched with saturated NH4Cl (10 mL) and extracted with EA (30 mL x 2). The organic layer was washed with brine (40 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (45 g silica gel, 0-25% EA in petroleum ether) to obtain 1-bromo-3-fluoro-8-methoxy-5,6,7,8-tetrahydronaphthalene-2-carboaldehyde (1300 mg, 53.3%) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 10.42 - 10.36 (m, 1H), 6.90 (d, J = 10.9 Hz, 1H), 4.54 (t, J = 2.7 Hz, 1H), 3.50 (s, 3H), 2.89 (dd, J = 17.8, 5.3 Hz, 1H), 2.81 - 2.66 (m, 1H), 2.46 - 2.35 (m, 1H), 1.99 - 1.86 (m, 1H), 1.80 - 1.69 (m, 1H), 1.58 - 1.44 (m, 1H). LCMS (ESI) m / z :287 [M+H] + .

[0536] Step 7: Synthesis of 4-bromo-5-methoxy-5,6,7,8-tetrahydro-1H-benzo[f]indazole (I-24g) To a solution of 1-bromo-3-fluoro-8-methoxy-5,6,7,8-tetrahydronaphthalene-2-carboaldehyde (1300 mg, 4.528 mmol) in DMSO (10 mL), hydrazine hydrate (2180 mg, 67.9 mmol) was added at 25 °C. The reaction mixture was then stirred at 125 °C for 12 hours. LC-MS showed that the desired product had been formed. The mixture was quenched with water (5 mL). The residue was extracted with EA (10 mL x 2). The combined organic phase was dried over Na2SO4, filtered, and concentrated. The residue was purified by flash column chromatography (25 g silica gel, 0-40% Â in petroleum ether) to obtain 4-bromo-5-methoxy-5,6,7,8-tetrahydro-1H-benzo[f]indazole (700 mg, 55%) as a white solid. 1 H NMR (400 MHz, CDCl3) δ 8.11 (s, 1H), 7.21 (s, 1H), 4.71 (t, J = 2.8 Hz, 1H), 3.49 (s, 3H), 3.10 - 2.99 (m, 1H), 2.89 - 2.78 (m, 1H), 2.39 (dd, J = 14.2, 3.8 Hz, 1H), 2.10 - 1.94 (m, 1H), 1.79 - 1.70 (m, 1H), 1.68 - 1.56 (m, 1H). LCMS (ESI) m / z 281[M+H] + .

[0537] Step 8: SFC chiral separation of 4-bromo-5-methoxy-5,6,7,8-tetrahydro-1H-benzo[f]indazole (I-24hi and I-24h-ii) 800 mg of the crude diastereoisomer mixture was purified by SFC (separation conditions: column: Daicel CHIRALCEL AD, 250 mm × 30 mm ID, 10 μm; mobile phase: CO2 / MeOH [0.2% NH3 (7 M solution in MeOH)] = 80 / 20; flow rate: 80 g / min; wavelength: UV 214 nm; temperature: 35 °C), and the unknown absolute configuration (R * Two isomers were obtained having (as shown). (R * )-4-bromo-5-methoxy-5,6,7,8-tetrahydro-1H-benzo[f]indazole (I-24h-i) (380 mg, 47.5%, Rt: 1.418 mins, peak 1) as a white solid. 1 H NMR (400 MHz, MeOD) δ 7.99 (s, 1H), 7.27 (s, 1H), 4.73 (s, 1H), 3.48 (d, J = 0.8 Hz, 3H), 3.04 (d, J = 16.5 Hz, 1H), 2.86 (ddd, J = 16.9, 11.3, 5.9 Hz, 1H), 2.46 - 2.37 (m, 1H), 1.95 (td, J = 12.6, 6.3 Hz, 1H), 1.80 - 1.70 (m, 1H), 1.68 - 1.58 (m, 1H). LCMS (ESI) m / z: 281 [M+H] + .[α] D 25 = + 5.50 (c=0.20, CH3OH). (R * )-4-bromo-5-methoxy-5,6,7,8-tetrahydro-1H-benzo[f]indazole (I-24h-ii) (400 mg, 50%, Rt: 1.731 min, peak 2) as a white solid. 1 H NMR (400 MHz, MeOD) δ 7.99 (s, 1H), 7.27 (s, 1H), 4.73 (s, 1H), 3.48 (s, 3H), 3.04 (d, J = 16.5 Hz, 1H), 2.91 - 2.80 (m, 1H), 2.46 - 2.37 (m, 1H), 2.02 - 1.88 (m, 1H), 1.79 - 1.71 (m, 1H), 1.69 - 1.58 (m, 1H). LCMS (ESI) m / z: 281 [M+H] + ,[α] D 25 = -5.50 (c=0.20, CH3OH).

[0538] Step 9: (R * Synthesis of )-4-bromo-5-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-5,6,7,8-tetrahydro-1H-benzo[f]indazole (I-24i and I-24ii) (R) in DMF (3 mL) * To a solution of )-4-bromo-5-methoxy-5,6,7,8-tetrahydro-1H-benzo[f]indazole (80 mg, 0.28 mmol), NaH (22.8 mg, 0.569 mmol) was added under N2 conditions at 0°C. The mixture was stirred at 0°C for 1 hour. Then, a solution of SEMCl (94.9 mg, 0.569 mmol) in DMF (1 mL) was added to the above solution at 0°C. The resulting mixture was stirred at 20°C for 1 hour. LC-MS showed that the starting material had been consumed and the desired product had been detected. The mixture was quenched with aqueous NH4Cl solution (0.5 mL) and concentrated under reduced pressure. The residue was purified by flash column chromatography (5 g silica gel, 0-20% siRNA in petroleum ether) and (R * )-4-bromo-5-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-5,6,7,8-tetrahydro-1H-benzo[f]indazole (100 mg, 85%) was obtained as an oily substance. 1 H NMR (400 MHz, CDCl3) δ 8.06 (s, 1H), 7.33 (s, 1H), 5.74 (s, 2H), 4.77 (t, J = 2.8 Hz, 1H), 3.61 - 3.50 (m, 5H), 3.16 - 3.07 (m, 1H), 2.95 - 2.84 (m, 1H), 2.47 - 2.39 (m, 1H), 2.06 (ddd, J = 8.9, 8.2, 2.7 Hz, 1H), 1.90 - 1.76 (m, 1H), 1.72 - 1.63 (m, 1H), 0.97 - 0.89 (m, 2H), 0.01 - -0.02 (m, 9H). LCMS (ESI) m / z 411 [M+H] + .

[0539] Synthesis of intermediate 25(I-25): ((7aS,Z)-2-(2-((tert-butyldimethylsilyl)oxy)propyridene)tetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol

[0540] [ka]

[0541] Step 1: Synthesis of (S)-5-oxo-2-(2-oxopropyridene)tetrahydro-1H-pyrrolidine-7a(5H)-carboxylate(I-25a) To a solution of ethyl(S)-2-methylene-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (1 g, 4.779 mmol) and (E)-penta-3-en-2-one (2010 mg, 23.9 mmol) in DMC (20 mL), 1,4-benzoquinone (52 mg, 0.478 mmol) and a second-generation Grubbs catalyst (406 mg, 0.478 mmol) were added. The mixture was then stirred at 40°C for 14 hours. LC-MS showed that the starting materials were consumed and the desired product was formed. The reaction mixture was filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel, petroleum ether with 0-100% ethyl) to obtain ethyl(S)-5-oxo-2-(2-oxopropyridene)tetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (250 mg, 21%) as a brown oily substance. 1 H NMR (400 MHz, CDCl3) δ 6.33 - 6.26 (m, 1H), 4.80 (d, J = 19.9 Hz, 1H), 4.20 (dd, J = 8.9, 5.4 Hz, 2H), 4.13 (d, J = 20.1 Hz, 1H), 3.21 (d, J = 17.3 Hz, 1H), 2.88 - 2.79 (m, 1H), 2.71 - 2.60 (m, 2H), 2.54 - 2.46 (m, 1H), 2.23 (s, 3H), 2.13 (dd, J = 10.1, 2.9 Hz, 1H), 1.26 (t, J = 7.1 Hz, 3H). LCMS (ESI) m / z: 252.1 [M+H] + .

[0542] Step 2: Synthesis of ethyl(S,Z)-5-oxo-2-(2-oxopropyridene)tetrahydro-1H-pyrrolidine-7a(5H)-carboxylate(I-25b) Ethyl(S)-5-oxo-2-(2-oxopropyridene)tetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (250 mg, 0.995 mmol) was purified by SFC (instrument: SFC80; column: CHIRALPAK IE-3 3 mm × 150 mm, 3 μm; flow rate: 80 g / min) at 35°C with elution at CO2 / MeOH [0.2% NH3 (7 M solution in MeOH)] = 70 / 30 to obtain ethyl(S,Z)-5-oxo-2-(2-oxopropyridene)tetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (75 mg, 30%) as a yellow oily substance. 1 H NMR (400 MHz, CDCl3) δ 6.29 (s, 1H), 4.81 (d, J = 19.9 Hz, 1H), 4.23 - 4.17 (m, 2H), 4.11 (s, 1H), 3.21 (d, J = 16.7 Hz, 1H), 2.93 - 2.78 (m, 1H), 2.73 - 2.57 (m, 2H), 2.56 - 2.44 (m, 1H), 2.22 (s, 3H), 2.18 - 2.09 (m, 1H), 1.26 (t, J = 7.1 Hz, 3H). LCMS (ESI) m / z: 252.2 [M+H] + .

[0543] Step 3: Synthesis of ethyl(7aS,Z)-2-(2-hydroxypropyridene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate(I-25c) To a solution of ethyl(S,Z)-5-oxo-2-(2-oxopropyridene)tetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (65 mg, 0.26 mmol) in methanol (3 mL), cerium(III) chloride (77 mg, 0.31 mmol), followed by NaBH4 (11 mg, 0.285 mmol), was added at 0°C. The reaction mixture was heated to 20°C and stirred for 15 minutes at 20°C. The reaction mixture was quenched with H2O (10 mL) and then extracted with DCM (20 mL x 5). The combined organic phase was washed with brine (20 mL), dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain crude ethyl(7aS,Z)-2-(2-hydroxypropylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (60 mg, 92%) as a colorless oil. LCMS(ESI) m / z: 254.2 [M+H] +

[0544] Step 4: Synthesis of ethyl(7aS,Z)-2-(2-((tert-butyldimethylsilyl)oxy)propyridene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate(I-25d) To a solution of ethyl(7aS,Z)-2-(2-hydroxypropylidene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (55 mg, 0.22 mmol) in DCM (3 mL), TBSCl (43 mg, 0.282 mmol), TEA (66 mg, 0.651 mmol), and DMAP (13 mg, 0.109 mmol) were added. The reaction mixture was stirred under N2 at 20°C for 14 hours. LC-MS showed that DP was formed. The reaction mixture was filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel, petroleum ether with 0-30% ethyl) to obtain ethyl(7aS,Z)-2-(2-((tert-butyldimethylsilyl)oxy)propyridene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (45 mg, 56%) as a yellow oil. 1H NMR (400 MHz, CDCl3) δ 5.52 - 5.31 (m, 1H), 4.27 (d, J = 14.6 Hz, 1H), 4.19 - 4.13 (m, 3H), 3.78 - 3.66 (m, 1H), 2.95 (dd, J = 22.8, 15.5 Hz, 1H), 2.79 - 2.69 (m, 1H), 2.63 - 2.53 (m, 1H), 2.49 - 2.38 (m, 2H), 2.09 (ddd, J = 9.6, 8.3, 2.9 Hz, 1H), 1.24 - 1.23 (m, 3H), 1.15 (dd, J = 6.2, 4.3 Hz, 3H), 0.84 (d, J = 2.3 Hz, 9H), 0.02 - -0.02 (m, 6H). LCMS (ESI) m / z: 390.13 [M +Na] + .

[0545] Step 5: Synthesis of ((7aS,Z)-2-(2-((tert-butyldimethylsilyl)oxy)propyridene)tetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol(I-25) To a solution of ethyl (7aS,Z)-2-(2-((tert-butyldimethylsilyl)oxy)propyridene)-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (40 mg, 0.11 mmol) in THF (2 mL), 1 M LiAlH4 (0.22 mL, 0.218 mmol) was added. The reaction mixture was stirred under N2 at 70°C for 1 hour. LC-MS showed that the starting material was consumed and the desired product was formed. Sodium sulfate decahydrate was added to the mixture and filtered. The filtrate was concentrated under reduced pressure to obtain crude ((7aS,Z)-2-(2-((tert-butyldimethylsilyl)oxy)propyridene)tetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol (35 mg) as a yellow oil. LC-MS (ESI) m / z: 312.2 [M+H] + .

[0546] Synthesis of intermediates 26i and 26ii (I-26i and I-26ii): [(2Z,7aS)-2-ethylidenetetrahydro-1H-pyrrolidine-7a(5H)-yl]methanol and [(2E,7aS)-2-ethylidenetetrahydro-1H-pyrrolidine-7a(5H)-yl]methanol

[0547] [ka]

[0548] Step 1: Synthesis of ethyl(7aS)-2-ethylidene-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (intermediate I-26a) Propylene (15 mL, 15.0 mmol) was added to a solution of ethyl(S)-2-methylene-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (CAS# 2820536-98-7, 627 mg, 3.00 mmol) and a second-generation Grubbs catalyst (254 mg, 0.300 mmol) in DCM (5 mL). The mixture was then stirred at 0°C for 20 hours. The solution was concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain ethyl(7aS)-2-ethylidene-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (intermediate I-26a, 230 mg, 34%) as a brown oil. 1 H NMR (400 MHz, DMSO) δ 5.43 - 539 (m, 1H), 4.16 - 3.98 (m, 3H), 3.66 - 3.48 (m, 1H), 3.06 - 2.81 (m, 1H), 2.60 - 2.56 (m, 1H), 2.45 - 2.14 (m, 4H), 1.61 - 1.49 (m, 3H), 1.23 - 1.14 (m, 3H). LCMS (ESI) m / z: 224.1

[0549] Step 2: Synthesis of ethyl(2Z,7aS)-2-ethylidene-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (intermediate I-26b) and ethyl(2E,7aS)-2-ethylidene-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (intermediate I-26c) Ethyl(7aS)-2-ethylidene-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (intermediate I-26a, 450 mg) was purified by chiral separation (instrument: SFC 150; column: Daicel CHIRALCEL AS, 250 mm × 30 mm ID, 10 μm; mobile phase: CO2 / MeOH [0.2% NH3 (7M solution in MeOH)] = 90 / 10; flow rate: 80 g / min; wavelength: UV 214 nm; temperature: 35 °C) to obtain (2Z,7aS)-2-ethylidene-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (intermediate I-26b, 180 mg, peak 1 of 2, Rt: 1.902 min, yield: 40%). 1 H NMR (400 MHz, CDCl3) δ 5.48 - 5.42 (m, 1H), 4.30 - 4.11 (m, 3H), 3.72 (d, J = 15.6 Hz, 1H), 3.00 (d, J = 15.3 Hz, 1H), 2.86 - 2.71 (m, 1H), 2.63 - 2.53 (m, 1H), 2.51 - 2.40 (m, 2H), 2.11 (ddd, J = 13.2, 10.8, 9.6 Hz, 1H), 1.63 - 1.55 (m, 3H), 1.31 - 1.23 (m, 3H) and ethyl(2E,7aS)-2-ethylidene-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (intermediate I-26c, 230 mg, peak 2 of 2, Rt: 2.415 min, yield: 51%) 1 H NMR (400 MHz, CDCl3) δ 5.49 - 5.39 (m, 1H), 4.32 - 4.15 (m, 3H), 3.69 (d, J = 15.0 Hz, 1H), 3.18 - 3.08 (m, 1H), 2.77 (dt, J = 16.5, 10.0 Hz, 1H), 2.62 (ddd, J = 13.0, 9.1, 1.4 Hz, 1H), 2.49 - 2.38 (m, 1H), 2.35 - 2.24 (m, 1H), 2.18 - 2.12 The values ​​obtained were (m, 1H), 1.67 - 1.58 and (m, 3H), 1.27 (t, J = 7.1 Hz, 3H).

[0550] Step 3: Synthesis of [(2Z,7aS)-2-ethylidenetetrahydro-1H-pyrrolidine-7a(5H)-yl]methanol (intermediate I-26i) To a solution of ethyl(2Z,7aS)-2-ethylidene-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (intermediate I-26b, 150 mg, 0.672 mmol) in THF (20 mL), LiAlH4 (1.3 mL, 1.3 mmol, 1 M in THF) was added at 20 °C under a nitrogen atmosphere. The resulting mixture was heated to 70 °C and stirred for 1 hour. The mixture was quenched with Na2SO4·10H2O (2 g), filtered, and concentrated under reduced pressure to obtain [(2Z,7aS)-2-ethylidenetetrahydro-1H-pyrrolidine-7a(5H)-yl]methanol (intermediate I-26i, 120 mg, 89%) as a colorless oil. 1 H NMR (400 MHz, CDCl3) δ 4.93 - 4.83 (m, 2H), 4.80 (d, J = 3.8 Hz, 1H), 4.21 (d, J = 3.2 Hz, 1H), 3.51 (d, J = 13.9 Hz, 1H), 3.34 (d, J = 13.9 Hz, 2H), 3.07 (dd, J = 9.8, 5.3 Hz, 1H), 3.02 (s, 2H), 2.46 (dd, J = 9.7, 8.4 Hz, 2H), 2.10 (dd, J = 12.8, 6.6 Hz, 1H), 1.52 (dd, J = 12.8, 6.3 Hz, 1H). LCMS (ESI) m / z: 168.3 [M+H] + .

[0551] Step 3': Synthesis of [(2E,7aS)-2-ethylidenetetrahydro-1H-pyrrolidine-7a(5H)-yl]methanol (intermediate I-26ii) To a solution of ethyl(2E,7aS)-2-ethylidene-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (intermediate I-26c, 180 mg, 0.806 mmol) in THF (20 mL), LiAlH4 (1.6 mL, 1.6 mmol, 1 M in THF) was added at 20°C under a nitrogen atmosphere. The resulting mixture was heated to 70°C and stirred for 1 hour. The mixture was quenched with Na2SO4·10H2O (2 g), filtered, and concentrated under reduced pressure. The crude mixture was purified by flash column chromatography (12 g silica gel, 0-20% MeOH in DCM (5% NH4OH in MeOH)) to obtain [(2Z,7aS)-2-ethylidenetetrahydro-1H-pyrrolidine-7a(5H)-yl]methanol (intermediate I-26ii, 70 mg, 41%) as a colorless oil. LCMS (ESI) m / z: 168.3 [M+H] + . [Examples]

[0552] Examples and general methods. Important preparation examples and procedures are illustrated to illustrate methods for synthesizing the examples.

[0553] (Example 1 (General Method A)) (8aS)-5-(6-chloro-5-cyclopropyl-1H-indazole-4-yl)-4-fluoro-2-{[(2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl]methoxy}-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene

[0554] [ka]

[0555] Preparation of Example 1:

[0556] [ka]

[0557] Step 1: Preparation of (8aS)-5-[6-chloro-5-cyclopropyl-2-(oxan-2-yl)-2H-indazole-4-yl]-4-fluoro-2-(methylsulfanyl)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene (1a). To a solution of (S)-4-fluoro-2-(methylthio)-5-(tributylstannyl)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxane (3 mL) and 4-bromo-6-chloro-5-cyclopropyl-2-(oxan-2-yl)-2H-indazole (I-5i, 172 mg, 0.482 mmol), CuI (91.9 mg, 0.482 mmol) and Pd(PPh3)4 (55.8 mg, 0.0482 mmol) were added. The reaction mixture was stirred under N2 at 110°C for 3 hours, concentrated, and the residue was purified by silica gel chromatography using a gradient of 0-100% siRNA in petroleum ether to separate 1a from 1b. (8aS)-5-[6-chloro-5-cyclopropyl-2-(oxan-2-yl)-2H-indazole-4-yl]-4-fluoro-2-(methylsulfanyl)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene (1a, 38 mg, 13%) was obtained as a yellow solid. 1H NMR (400 MHz, CDCl3) δ 7.86 (dd, J = 28.4, 5.9 Hz, 1H), 7.77 (d, J = 17.1 Hz, 1H), 5.69 (dd, J = 18.8, 7.1 Hz, 1H), 5.29 - 5.17 (m, 1H), 4.60 (dd, J = 36.6, 20.6 Hz, 2H), 4.22 (d, J = 11.4 Hz, 2H), 4.03 (s, 2H), 3.91 - 3.67 (m, 2H), 3.49 (dd, J = 22.9, 9.5 Hz, 2H), 2.62 (d, J = 5.4 Hz, 2H), 2.24 (s, 1H), 2.05 (s, 3H), 1.83 - 1.61 (m, 3H), 0.89 (ddd, J = 17.5, 11.4, 6.3 Hz, 1H), 0.69 (s, 1H), 0.33 - 0.08 (m, 2H), MS: 597 [M+H]+.(8aS)-5-(6-chloro-5-cyclopropyl-2H-indazole-4-yl)-4-fluoro-2-(methylsulfanyl)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene (1b, 45 mg, 18%) was also recovered as a yellow solid, but it was not included in the next step.

[0558] Step 2: Preparation of (8aS)-5-[6-chloro-5-cyclopropyl-2-(oxan-2-yl)-2H-indazole-4-yl]-4-fluoro-2-{[(2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl]methoxy}-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene(1c) To a solution of (8aS)-5-[6-chloro-5-cyclopropyl-2-(oxan-2-yl)-2H-indazole-4-yl]-4-fluoro-2-(methylsulfanyl)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene (1a, 38 mg, 0.046 mmol), mCPBA (12 mg, 0.069 mmol) was added. The reaction mixture was stirred at room temperature for 2 hours. LC-MS analysis showed that the starting material was consumed and the sulfoxide was detected. The mixture was quenched with aqueous Na2SO3 (10 mL) and diluted with DCM (20 mL). The organic layer was washed with aqueous NaHCO3 (20 mL), followed by saturated aqueous NaCl (20 mL). After drying and concentrating on NaSO4, 32 mg of sulfoxide was converted into a yellow solid, MS 613 [M+H]. + The following was obtained: A solution of sulfoxide (32 mg, 0.052 mmol) and ((2S,7aR)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol, CAS 2097518-76-6 (16.6 mg, 0.104 mmol) in DCM (2 mL) was cooled to 0°C. LiHMDS (1 M 0.0626 mL, 0.0626 mmol) was added, and the mixture was stirred at 25°C for 1 hour. LCMS analysis showed product formation. The reaction mixture was extracted with EA (2 × 20 mL), and the combined organic extract was washed with water (10 mL). After concentration, the residue was purified by preparative TLC using DCM / MeOH 10 / 1 to obtain (8aS)-5-[6-chloro-5-cyclopropyl-2-(oxan-2-yl)-2H-indazole-4-yl]-4-fluoro-2-{[(2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl]methoxy}8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene (1c, 20 mg) as a yellow solid, MS:708 [M+H]. + It was obtained as such.

[0559] Step 3: Synthesis of (8aS)-5-(6-chloro-5-cyclopropyl-1H-indazole-4-yl)-4-fluoro-2-{[(2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl]methoxy}-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene, Example 1. A solution of (8aS)-5-[6-chloro-5-cyclopropyl-2-(oxan-2-yl)-2H-indazole-4-yl]-4-fluoro-2-{[(2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl]methoxy}-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene (1c, 15 mg, 0.021 mmol) in DCM (1 mL) was treated with HCl (4 M, 0.5 mL, 2.0 mmol) in 1,4-dioxane, and the mixture was stirred at room temperature for 1 hour. The mixture was concentrated, and the residue was purified by preparative HPLC to obtain Example 1 as a white solid (5.5 mg). 1 ¹H NMR (400 MHz, MeOD) δ 8.53 (s, 1H), 7.87 and 7.81 (2 s, major and minor rotational isomers, 1H), 7.75 (s, 1H), 5.38 (d, J = 53.6 Hz, 1H), 5.24 (dtd, J = 9.4, 6.8, 2.8 Hz, 1H), 4.76 (ddd, J = 13.4, 4.1, 2.1 Hz, 1H), 4.61 (t, J = 13.0 Hz, 1H), 4.50 - 4.28 (m, 3H), 4.26 - 4.15 (m, 1H), 4.00 (dt, J = 10.9, 5.1 Hz, 1H), 3.83 (ddd, J = 34.2, 12.3, 10.0 Hz, 1H), 3.64 - 3.55 (m, 1H), 3.50 - 3.37 (m, 4H), 3.19 - 3.08 (m, 1H), 2.51 - 2.28 (m, 2H), 2.28 - 2.15 (m, 2H), 2.14 - 1.89 (m, 5H), 0.99 - 0.81 (m, 1H), 0.74 - 0.62 (m, 1H), 0.34 - 0.10 (m, 2H), MS 624 [M+H] + .

[0560] (Example 24 (General Method B)) (8aS)-5-[6-chloro-5-(propan-2-yl)-1H-indazole-4-yl]-4-fluoro-2-{[(7aS)-2-methylidenetetrahydro-1H-pyrrolidine-7a(5H)-yl]methoxy}-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene

[0561] [ka] Example 24 was prepared in the same manner as in Example 1, however, the one-pot Suzuki coupling method was used instead of Stannan chemistry. The one-pot procedure is illustrated in General Scheme 2 (Method B).

[0562] [ka]

[0563] Synthesis of (8aS)-5-(6-chloro-5-isopropyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-4-yl)-4-fluoro-2-(methylthio)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene (I-24a) To a solution of 4-bromo-6-chloro-5-isopropyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole (I-6, 45.3 mg, 0.127 mmol) in DMF (3.2 mL), (S)-5-chloro-4-fluoro-2-(methylthio)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene (I-2, 40 mg, 0.11 mmol), bis(pinacorato)diborone (41.9 mg, 0.165 mmol), CsF (66.8 mg, 0.44 mmol), and cataCXium A Pd G3 (8.16 mg, 0.0112 mmol) were added at 25°C under nitrogen. The resulting mixture was stirred at 70°C for 20 hours. LC-MS detected the desired product, indicating that the starting material had been consumed. The mixture was concentrated under vacuum. The residue was purified by flash column chromatography (Combi-Flash, 12 g silica gel, 0-100% siRNA in petroleum ether) to obtain the title intermediate (45 mg, 59%) as a yellow solid. 1 ¹H NMR (400 MHz, CDCl3) δ 7.76 and 7.72 (2 s, 1H, major and minor rotational isomers, respectively), 7.59 - 7.46 (m, 1H), 5.73 - 5.62 (m, 1H), 5.31 - 5.19 (m, 1H), 4.74 - 4.47 (m, 2H), 4.31 - 4.13 (m, 2H), 4.07 - 3.96 (m, 2H), 3.85 - 3.70 (m, 2H), 3.56 - 3.14 (m, 3H), 2.63 (s, 3H), 2.52 - 2.43 (m, 1H), 2.31 - 2.11 (m, 3H), 1.78 - 1.65 (m, 4H), 1.47 - 1.39 (m, 3H), 1.37 - 1.29 (m, 3H). 19 F NMR (376 MHz, CDCl3) δ -140.59. MS:599.2 [M+H] + .

[0564] Compound I-24a was converted to Example 24 by General Method A, which involves non-material modifications and / or substitutions, as outlined and illustrated in Example 1. See Table 1 for characterization of the examples.

[0565] (Example 40 (General Method C)) (8aS)-5-(6-chloro-5-cyclopropyl-1H-indazole-4-yl)-2-{[(7aS)-2-cyclopropyridenetetrahydro-1H-pyrrolidine-7a(5H)-yl]methoxy}4-fluoro-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene

[0566] [ka] Similar to Example 1, but using the Suzuki coupling method with an indazole boronate derivative, Example 40 was prepared. The procedure is illustrated in General Scheme 3 (Method c).

[0567] [ka]

[0568] Synthesis of (8aS)-5-(6-chloro-5-cyclopropyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-4-yl)-4-fluoro-2-(methylthio)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene In a 100 mL vial, add 6-chloro-5-cyclopropyl-1-(tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole (I-20, 4.38 g, 9.8 mmol, 1 equivalent), (S)-5-chloro-4-fluoro-2-(methylthio)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3, 6,13a-tetraazanaphtho[1,8-ab]heptalene (3.84 g, 10.8 mmol, 1.1 equivalents), Pd(OAc)2 (137 mg, 0.6 mmol, 0.0625 equivalents), CsF (5.95 g, 39.2 mmol, 4 equivalents), Ph2P(2-MeO)Ph (358 mg, 1.22 mmol, 0.125 equivalents), water (1760 mg, 97.9 mmol, 10 equivalents), and t-amyl OH (50 mL) were charged. The reaction mixture was purged three times with nitrogen and heated at 70°C under nitrogen for 16 hours. LC-MS showed that most of the starting material was consumed and a major peak with the desired MS was detected. The reaction mixture was filtered. The filtrate was diluted with water (200 mL) and extracted with CH2Cl2 (3 × 100 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (120 silica gel, 0-25% ethyl phosphate in DCM) to obtain (8aS)-5-(6-chloro-5-cyclopropyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-4-yl)-4-fluoro-2-(methylthio)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene (40a, 1.8g, 31%) as a gray solid. 1H NMR (400 MHz, MeOD) δ 7.94 - 7.89 (m, 1H), 7.85 and 7.80 (2 s, 1H, major and minor isomers, respectively), 5.90 - 5.74 (m, 1H), 5.32 - 5.20 (m, 1H), 4.78 - 4.69 (m, 1H), 4.67 - 4.55 (m, 1H), 4.38 - 4.26 (m, 1H), 4.21 and 4.19 (2 dd, J = 12.6, 4.2 Hz, 1H, principal and secondary isomers, respectively), 3.99 (dt, J = 12.5, 5.1 Hz, 2H), 3.92 - 3.73 (m, 2H), 3.62 - 3.53 (m, 1H), 3.51 - 3.38 (m, 1H), 2.62 (s, 3H), 2.51 - 2.38 (m, 1H), 2.28 - 1.95 (m, 5H), 1.90 - 1.61 (m, 3H), 0.97 - 0.83 (m, 1H), 0.78 - 0.65 (m, 1H), 0.34 - 0.09 (m, 2H). LCMS (ESI) m / z: 597.3 (M+H) +.

[0569] Compound 40a was converted to Example 40 by General Method A, which involves non-material modifications and / or substitutions, as outlined and illustrated in Example 1. See Table 1 for characterization of the examples.

[0570] (Example 46 (General Method D)) (8aS)-5-(3,6-dichloro-5-cyclopropyl-1H-indazole-4-yl)-4-fluoro-2-{[(7aS)-2-methylidenetetrahydro-1H-pyrrolidine-7a(5H)-yl]methoxy}8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene

[0571] [ka] Example 46 was prepared in the same manner as Example 1, however, chlorination and oxidation were achieved by treating the indazole precursor with N-chlorosuccinimide. The procedure is illustrated in General Scheme 4 (Method D).

[0572] [ka] (8aS)-5-(6-chloro-5-cyclopropyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-4-yl)-4-fluoro-2-(methylthio)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene intermediate (40a, 300 mg, 0.50 mmol) was dissolved in DMF (2.5 mL). N-chlorosuccinamide (168 mg, 1.26 mmol) was added, and the reaction mixture was stirred at room temperature for 3 hours. By LC-MS, a mixture of products was obtained. The reaction mixture was diluted with Depositphotos (30 mL) and water (5 mL). After separation of the layers, the organic extract was washed with water (3 × 10 mL) to remove all DMF. After drying over Na2SO4, the extract was concentrated, and the crude was placed in the next step without purification. The crude from the chlorination step and (S)-(2-methylenetetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol (97.0 mg, 0.63 mmol) were dissolved in CH3CN (4.52 mL). The solution was cooled to 0°C, and LiOtBu (2.2 M, 0.31 mL, 0.68 mmol) was rapidly added over 30 seconds to obtain a suspension. After stirring for 15 minutes, the reaction mixture was removed from the ice bath and stirred at room temperature for 20 minutes. LCMS analysis showed that THP-protected Example 46 had been formed, and one drop of HOAc was added. After stirring for 5 minutes, TFA (2 mL) was added to the mixture, and the reaction mixture was stirred at room temperature for 16 hours. The mixture was heated to 50°C for 4 hours until partial deprotection occurred and the deprotection of the THP group was complete. The mixture was concentrated and preparatively purified by flash chromatography with elution under a gradient of 0–10% MeOH in DCM. The pure fraction was collected and concentrated to obtain a white foam, which was further purified using HPLC (Phenemonex Gemini NX C18, 150 × 21.2 mm, 5 μm, AXIA Pack; mobile phase A: water + 10 mM ammonium acetate, mobile phase B: 20–70% acetonitrile B, 40 mL / min at 8.0 min). Evidence of atropisomers was observed on a chiral column, and attempts to separate the atropisomers resulted in equilibration back into the mixture at room temperature.(8aS)-5-(3,6-dichloro-5-cyclopropyl-1H-indazole-4-yl)-4-fluoro-2-{[(7aS)-2-methylidenetetrahydro-1H-pyrrolidine-7a(5H)-yl]methoxy}8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene (Example 46) was presented as a single compound (19 mg, 5.8%). 1 H NMR (400 MHz, DMSO-d6) δ 7.82 (s, 1H), 7.33 (d, J = 4.3 Hz, 1H), 5.09 - 4.95 (m, 1H), 4.92 (d, J = 3.7 Hz, 2H), 4.66 (dd, J = 13.1, 4.5 Hz, 1H), 4.55 (dd, J = 13.2, 7.0 Hz, 1H), 4.33 (dt, J = 9.8, 5.1 Hz, 1H), 4.09 (ddd, J = 16.6, 8.4, 4.7 Hz, 4H), 3.95 - 3.83 (m, 1H), 3.66 - 3.55 (m, 3H), 3.03 (dt, J = 10.7, 5.2 Hz, 2H), 2.69 - 2.53 (m, 2H), 2.42 - 2.28 (m, 2H), 2.10 - 1.94 (m, 3H), 1.84 - 1.63 (m, 3H), 0.88 - 0.57 (m, 2H), 0.37 - 0.12 (m, 2H); LCMS m / z: 652.2 [M+H] + .

[0573] (Example 3 (General Method E)) (8aS)-4-fluoro-2-{[(2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl]methoxy}5-(1H-indazole-4-yl)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene

[0574] [ka] Example 3 was prepared according to General Scheme 5 (Method E).

[0575] [ka]

[0576] Step 1: Synthesis of (8aS)-5-chloro-4-fluoro-2-{[(2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl]methoxy}-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene(3b) To a solution of crude (8aS)-5-chloro-4-fluoro-2-(methanesulfonyl)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene (3.2 g, 8.97 mmol, 1 equivalent) in acetonitrile (29.9 mL, 0.3 M), ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol, CAS# 2097518-76-6 (1.71 g, 10.7 mmol), followed by lithium trimethylsilanolate (2.5 g, 26.8 mmol) was added. The mixture was heated to 60°C until complete conversion was achieved, then cooled to 0°C in an ice bath and quenched with water (100 mL). The aqueous mixture was extracted with toluene (3 × 100 mL), the combined organic layer was washed with brine, dried over sodium sulfate, filtered, and concentrated to obtain the desired product (3.88 g, 92% in two steps) as a solid, which was used crudely without purification.

[0577] Step 2: Synthesis of (8aS)-4-fluoro-2-{[(2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl]methoxy}5-(1H-indazole-4-yl)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene (Example 3). (8aS)-5-chloro-4-fluoro-2-{[(2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl]methoxy}8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene (70 mg, 0.15 mmol, 1.0 equivalent), 1H-indazole-4-boronic acid (31.5 mg, 0.194 mmol, 1.3 equivalents), RuPhos PdG4 (25.4 mg, 0.03 mmol) A mixture of 0.2 equivalents of 0.45 mmol of K₂CO₃ (62.0 mg, 0.45 mmol, 3.0 equivalents) was dissolved in dioxane (0.748 mL) and H₂O (0.0748 mL). The mixture was then degassed by blowing nitrogen into it for 5 minutes while stirring. The reaction container was capped and heated to 90°C until the reaction was complete. The resulting slurry was cooled to room temperature, at which point H₂O (2 mL) was added dropwise. The slurry was filtered, rinsed with H₂O, and the solid was purified by chromatography to obtain the title product (24.5 mg) as a solid. 1 H NMR (400 MHz, DMSO) δ 13.16 (s, 1H), 8.26 (d, J = 1.5 Hz, 1H), 7.61 (d, J = 8.1 Hz, 1H), 7.59 - 7.54 (m, 1H), 7.43 (dd, J = 8.3, 7.2 Hz, 1H), 5.34 - 5.12 (m, 1H), 4.93 (ddd, J = 13.7, 6.4, 3.3 Hz, 1H), 4.64 (dd, J = 13.4, 4.4 Hz, 1H), 4.47 (d, J = 13.2 Hz, 1H), 4.24 (dt, J = 9.1, 4.2 Hz, 1H), 4.11 (d, J = 10.3 Hz, 1H), 4.06 (dd, J = 12.3, 4.0 Hz, 1H), 4.03 - 3.98 (m, 1H), 3.82 (dt, J = 12.6, 5.0 Hz, 1H), 3.59 (dd, J = 12.3, 9.8 Hz, 1H), 3.34 (dddd, J = 28.2, 13.9, 9.1, 4.7 Hz, 2H), 3.10 (d, J = 5.3 Hz, 1H), 3.06 - 3.00 (m, 2H), 2.96 (d, J = 2.2 Hz, 1H), 2.77 (q, J = 8.1 Hz, 1H), 2.10 - 1.89 (m, 4H), 1.88 - 1.66 (m, 4H). 19F NMR (377 MHz, DMSO) δ -145.28, -171.96, -171.99, -172.05, -172.10, -172.12, -172.15, -172.20, -172.22, -172.26, -172.29.

[0578] Examples 56 and 57 were prepared according to general scheme A; however, an additional deprotection step was required to obtain the product (scheme A+):

[0579] [ka]

[0580] (Example 56) (8aS)-4-fluoro-5-((R *Synthesis of )-5-methoxy-5,6,7,8-tetrahydro-1H-benzo[f]indazole-4-yl)-2-(((S)-2-methylenetetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene (8aS)-4-fluoro-5-((R) in TBAF (0.858 mL, 0.858 mmol) * A solution of )-5-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-5,6,7,8-tetrahydro-1H-benzo[f]indazole-4-yl)-2-(((S)-2-methylenetetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene (obtained by coupling intermediate I-24i or I-24ii with intermediate I-4 according to Scheme A and Example 1 (50 mg, 0.066 mmol)) was stirred at 50°C for 3 hours. LCMS showed that the starting material was consumed and the desired product was detected, and the reaction mixture was subjected to preparative HPLC (column: WELCH Xtimate C18 21.2×250 mm). The product was purified using 10 μm; mobile phase: CH3CN-H2O (0.1% NH4HCO3); gradient: 30% CH3CN to 100% CH3CN; flow rate: 30 mL / min) to obtain (8aS)-4-fluoro-5-((R*)-5-methoxy-5,6,7,8-tetrahydro-1H-benzo[f]indazole-4-yl)-2-(((S)-2-methylenetetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene (9.04 mg, Y: 22%) as a white solid. 1¹H NMR (400 MHz, MeOD) δ 7.78 and 7.68 (2 s, 1H, sub-rotational isomer and major rotational isomer, respectively), 7.42 (s, 1H), 5.29 - 5.18 (m, 1H), 5.00 (s, 2H), 4.86 - 4.71 (m, 2H), 4.63 and 4.59 (2 d, J = 13.1 Hz, 1H, sub-rotational isomer and principal rotational isomer, respectively), 4.59 (d, J = 13.1 Hz, 1H), 4.40 - 4.29 (m, 3H), 4.18 (dd, J = 12.4, 4.0 Hz, 1H), 4.04 - 3.95 (m, 1H), 3.85 - 3.71 (m, 2H), 3.61 - 3.52 (m, 1H), 3.51 - 3.40 (m, 1H), 3.39 - 3.35 (m, 1H), 3.21 - 3.07 (m, 2H), 2.98 - 2.85 (m, 4H), 2.81 (d, J = 16.3 Hz, 1H), 2.77 - 2.70 (m, 1H), 2.50 (d, J = 15.8 Hz, 1H), 2.24 - 2.13 (m, 3H), 2.05 - 1.83 (m, 6H), 1.80 - 1.67 (m, 1H).LCMS (ESI) m / z 628 [M+1] + .

[0581] (Example 62) (8aS)-5-[(5R)-3-chloro-5-methyl-5,6,7,8-tetrahydro-1H-benzo[f]indazole-4-yl]-4-fluoro-2-{[(7aS)-2-methylidenetetrahydro-1H-pyrrolidine-7a(5H)-yl]methoxy}-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene

[0582] [ka] A solution of (8aS)-4-fluoro-2-{[(7aS)-2-methylidenetetrahydro-1H-pyrrolidine-7a(5H)-yl]methoxy}-5-[(5R)-5-methyl-5,6,7,8-tetrahydro-1H-benzo[f]indazole-4-yl]-8a,9,12,13-tetrahydro-8H,11H-7,10-dioxa-1,3,6,13a-tetraazanaphtho[1,8-ab]heptalene (Example 29, 50 mg, 0.049 mmol) and N-chlorosuccinimide (13 mg, 0.098 mmol) was mixed with a 3M potassium hydroxide solution in water (0.065 mL, 0.2 mmol). The mixture was stirred for 14 hours. LC-MS showed that the starting material was consumed and the desired product was formed as a mixture of two atropisomers. The reaction mixture was diluted with ethyl acetate (6 mL). It was washed with water (5 mL) and brine (5 mL), then dried over sodium sulfate and concentrated under reduced pressure. The residue was separated by preparative HPLC (Waters XSELECT CSH Prep C18). OBD, 100 × 30 mm, 5 μm. Mobile phase A: Water + 10 mM ammonium acetate. Mobile phase B: Acetonitrile. Purified over 8.5 minutes by gradient from 35% B to 100% B, (8aS)-5-[(5R)-3-chloro-5-methyl-5,6,7,8-tetrahydro-1H-benzo[f]indazole-4-y...

Claims

1. Compound of formula (III): 【Chemistry 1】 or a pharmaceutically acceptable salt thereof [In the formula, R １ is C ３ -C １０ cycloalkyl or a 4- to 12-member heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from the group consisting of N, O, and S, and the C ３ -C １０ cycloalkyl or 4- to 12-member heterocycloalkyl is each independently substituted with 1, 2 or 3 substituents selected from the group consisting of -OH, -CN, halogen, C １ -C ３ alkylidene, C １ -C ３ haloalkylidene, C １ -C ３ alkyl (wherein, when present, two of the C １ -C ３ alkyl may together with the carbon to which they are attached form a spirocyclic ring), C １ -C ３ alkoxy, -OC(O)NH ２ -, -OC(O)NHCH ３ -, -OC(O)N(CH ３ ) ２ and may be independently substituted with 1, 2 or 3 substituents selected from the group consisting of, and the C １ -C ３ alkyl, C １ -C ３ alkoxy, C １ -C ３ haloalkylidene or C １ -C ３ alkylidene may each be further substituted with 1, 2 or 3 R １１ . R ３ teeth, 【Chemistry 2】 And, R ３ is 1, 2, or 3 R １０ It may also be replaced with L is L1-L2-L3, and each of L1, L2, and L3 is a bond, -O(CH ２ ) ｎ - (where n is 0 or 1), -S-, -NR ７ -, and -CR ８ R ９ —are independently selected from the group consisting of L1, L2, and L3, provided that at least one of L1, L2, and L3 is not a combination. R ７ , R ８ , and R ９ Each of these is independently H or C １ ~C ３ It is alkyl, R １０ is, -NH ２ , halogen, -CN, C １ ~C ３ Alkyl, C １ ~C ３ Fluoroalkyl, C １ ~C ３ Alkoxy, C ２ ~C ３ Alkinyl, C ３ ~C ５ The C is independently selected from the group consisting of cycloalkyls and 4- to 6-membered heterocycloalkyls containing one or two heteroatoms selected from the group consisting of N, O, and S, and １ ~C ３ Alkyl, C ３ ~C ５ Cycloalkyl or 4- to 6-membered heterocycloalkyl groups are -OH, C １ ~C ３ Alkoxy, C １ ~C ３ It may be further substituted with one or two substituents independently selected from the group consisting of alkyls and halogens, or with two R １０ Along with the C atom to which they are bonded, there are 1, 2, or 3 R atoms. １２ C may be further replaced by ３ ~C ６ It may also form a cycloalkyl ring. R １１ -CN, -OH, methyl, -OCH ３ , C １ ~C ３ Alkoxy,-cyclopropyl,-oxetane,-C(O)NR ７ R ８ , -SO ２ R ９ and halogens are independently selected from the group, or two R １１ C ３ ~C ６ It forms a cycloalkyl ring or a 3-6 membered heterocycloalkyl ring, R １２ is -OH, C １ ~C ３ Alkoxy, -C １ ~C ３ [Selected independently from the group consisting of alkyls and halogens.]

2. The linker L is -(O-CH ２ ) - the compound according to claim 1, or a pharmaceutically acceptable salt thereof.

3. R １ is a 5- to 10-membered heterocycloalkyl containing 1 or 2 heteroatoms selected from the group consisting of N and O, and the 5- to 10-membered heterocycloalkyl is —OH, —CN, halogen, C １ ~C ３ alkylidenyl, C １ ~C ３ haloalkylidenyl, C １ ~C ３ alkyl (wherein, when present, two of the C １ ~C ３ alkyl may together with the carbon to which they are attached form a spirocyclic ring), C １ ~C ３ alkoxy, —OC(O)NH ２ , —OC(O)NHCH ３ , —OC(O)N(CH ３ )([[]] ２ [[]]) ２ and may be independently substituted with 1, 2 or 3 substituents selected from the group consisting of, and the C １ ~C ３ alkyl, C １ ~C ３ alkoxy, C １ ~C ３ haloalkylidenyl or C １ ~C ３ alkylidenyl may each be further substituted with 1, 2 or 3 R １１ substituents, the compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof.

4. L-R １ However, -OH, -CN, halogen, C １ ~C ３ Alkyridenyl, C １ ~C ３ Haloalkylidenyl, C １ ~C ３ Alkyl (wherein C if present) １ ~C ３ Two of the alkyl groups may form a spirocyclic ring together with the carbon atoms to which they are bonded. １ ~C ３ Alkoxy, -OC(O)NH ２ , -OC(O)NHCH ３ , -OC(O)N(CH ３ ) ２ It may be substituted with one, two, or three substituents independently selected from the group consisting of the following: 【Transformation 3】 And the above C １ ~C ３ Alkyl, C １ ~C ３ Alkoxy, C １ ~C ３ Haloalkylidenyl or C １ ~C ３ Alkylidenyl has 1, 2, or 3 R １１ The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, which may be further substituted with substituents.

5. L-R １ However, it may be substituted with one, two, or three halogens. 【Chemistry 4】 The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof.

6. L-R １ However, -OH, -CN, halogen, C １ ~C ３ It may be substituted with one or two substituents independently selected from the group consisting of alkyl groups. 【Transformation 5】 And the above C １ ~C ３ Alkylidenyl has 1 or 2 R １１ The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, which may be further substituted with substituents.

7. L-R １ However, -OH, -CN, halogen, C １ ~C ３ It may be substituted with one or two substituents independently selected from the group consisting of alkyl groups. 【Transformation 6】 And R １１ However, H, -CN, -OH, methyl, -OCH ３ , C １ ~C ３ Alkoxy,-cyclopropyl,-oxetane,-C(O)NR ７ R ８ , -SO ２ R ９ and halogens are independently selected from the group, or the R １１ However, along with the carbon atoms to which they are bonded, C ３ ~C ６ A compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, which forms a cycloalkyl ring or a 3- to 6-membered heterocycloalkyl ring.

8. L-R １ However, -OH, -CN, halogen, C １ ~C ３ Alkyridenyl, C １ ~C ３ Haloalkylidenyl and C １ ~C ３ It may be substituted with one, two, or three substituents independently selected from the group consisting of alkyl groups. 【Transformation 7】 The compound according to claim 7, or a pharmaceutically acceptable salt thereof.

9. L-R １ but, 【Transformation 8】 A compound according to claim 4, or a pharmaceutically acceptable salt thereof, selected from the group consisting of the above.

10. L-R １ but, 【Chemistry 9】 A compound according to claim 4, or a pharmaceutically acceptable salt thereof, selected from the group consisting of the above.

11. L-R １ but, 【Chemistry 10】 A compound according to claim 4, or a pharmaceutically acceptable salt thereof, selected from the group consisting of the above.

12. L-R １ but, 【Chemistry 11】 A compound according to claim 4, or a pharmaceutically acceptable salt thereof, selected from the group consisting of the above.

13. R ３ However, 1, 2, or 3 R １０ It may be replaced with 【Chemistry 12】 The compound according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof.

14. R ３ but, 【Chemistry 13】 The compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof.

15. R ３ but, 【Chemistry 14】 And, R １０ However, -NH ２ , halogen, -CN, C １ ~C ３ Alkyl, C １ ~C ３ Fluoroalkyl, C １ ~C ３ Alkoxy, C ２ ~C ３ Alkinyl, C ３ ~C ５ The C is independently selected from the group consisting of cycloalkyls and 4- to 6-membered heterocycloalkyls containing one or two heteroatoms selected from the group consisting of N, O, and S, and １ ~C ３ Alkyl, C ３ ~C ５ Cycloalkyl or 4-6 member heterocycloalkyl groups -OH, C １ ~C ３ Alkoxy, -C １ ~C ３ A compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, which may be further substituted with one or two substituents independently selected from the group consisting of alkyls and halogens.

16. R ３ but, 【Chemistry 15】 The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof.

17. R ３ but, 【Chemistry 16】 The compound according to claim 15, or a pharmaceutically acceptable salt thereof.

18. R ３ However, one R １０ Or one, two, or three R １２ It may be replaced with 【Chemistry 17】 The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof.

19. R ３ but, [Chemistry 18] The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof.

20. R ３ but, 【Chemistry 19】 The compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof.

21. R ３ but, 【Chemistry 20】 The compound according to claim 13, or a pharmaceutically acceptable salt thereof.

22. R ３ but, 【Chemistry 21】 The compound according to claim 20, or a pharmaceutically acceptable salt thereof.

23. R ３ but, 【Chemistry 22】 The compound according to claim 20, or a pharmaceutically acceptable salt thereof.

24. R ３ but, 【Chemistry 23】 The compound according to claim 20, or a pharmaceutically acceptable salt thereof.

25. R ３ but, 【Chemistry 24】 The compound according to claim 20, or a pharmaceutically acceptable salt thereof.

26. R ３ but, 【Chemistry 25】 The compound according to claim 20, or a pharmaceutically acceptable salt thereof.

27. R ３ but, 【Chemistry 26】 A compound according to any one of claims 1 to 14, selected from the group consisting of the above, or a pharmaceutically acceptable salt thereof.

28. R ３ but, 【Chemistry 27】 A compound according to any one of claims 1 to 13 or 15, selected from the group consisting of the above, or a pharmaceutically acceptable salt thereof. 【Request Item 29】 【Chemistry 28】 A compound or a pharmaceutically acceptable salt thereof, selected from the group consisting of the following. 【Request Item 30】 【Chemistry 29】 A compound or a pharmaceutically acceptable salt thereof, selected from the group consisting of the following. 【Request Item 31】 【Chemistry 30】 A compound or a pharmaceutically acceptable salt thereof, selected from the group consisting of the following. 【Request Item 32】 【Chemistry 31】 A compound or a pharmaceutically acceptable salt thereof, selected from the group consisting of the following. 【Request Item 33】 【Chemistry 32】 A compound selected from the group consisting of the following, or a pharmaceutically acceptable salt thereof.

34. A pharmaceutical composition comprising a compound according to any one of claims 1 to 33, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable additive.

35. A method for treating cancer, comprising administering a therapeutically effective amount of a compound according to any one of claims 1 to 33, or a pharmaceutically acceptable salt thereof, to a subject in need thereof.

36. A method for treating cancer, comprising administering a therapeutically effective amount of a compound according to any one of claims 1 to 33, or a pharmaceutically acceptable salt thereof, as a single agent to a subject in need thereof.

37. A method for treating cancer, comprising administering a therapeutically effective amount of a compound according to any one of claims 1 to 33, or a pharmaceutically acceptable salt thereof, to a subject in need thereof, and further comprising administering a therapeutically effective amount of an additional anticancer drug.

38. A method for treating cancer according to any one of claims 1 to 33, wherein the cancer is small cell lung cancer (NSCLC), pancreatic cancer, or colorectal cancer.

39. A compound according to any one of claims 1 to 33, or a pharmaceutically acceptable salt thereof, for use as a pharmaceutical.

40. A compound according to any one of claims 1 to 33, or a pharmaceutically acceptable salt thereof, for use in the treatment of cancer.

41. A compound or a pharmaceutically acceptable salt thereof for use in the treatment of a cancer according to any one of claims 1 to 33, wherein the cancer is small cell lung cancer (NSCLC), pancreatic cancer, or colorectal cancer.

42. Use of a compound according to any one of claims 1 to 33, or a pharmaceutically acceptable salt thereof, for the manufacture of a pharmacopoeia for treating cancer.

43. A method for treating a disorder mediated by inhibition of KRAS G12C, KRAS G12D, and KRAS G12V receptors in a subject, comprising administering to a subject in need of such treatment a compound according to any one of claims 1 to 33, or a pharmaceutically acceptable salt thereof, in an amount effective for treating the disorder.

44. A pharmaceutical combination comprising a compound according to any one of claims 1 to 33, or a pharmaceutically acceptable salt thereof, and at least one additional therapeutic agent or a pharmaceutically acceptable salt thereof, wherein the combination is fixed or unfixed.

45. A pharmaceutical composition comprising the pharmaceutical combination described in claim 44 and at least one additive.

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