Compounds and uses thereof

Abstract

The present disclosure features compounds for treating BAF complex-associated disorders.

Description

[0001] background

[0002] This invention relates to compounds for regulating the BRG1 or BRM-associated factor (BAF) complex. In particular, this invention relates to compounds that can be used to treat conditions associated with BAF complex function.

[0003] Chromatin regulation is essential for gene expression, and ATP-dependent chromatin remodeling is the mechanism by which this gene expression occurs. The human switch / sucrose non-fermentable (SWI / SNF) chromatin remodeling complex, also known as the BAF complex, contains two SWI2-like ATPases called BRG1 (Brahma-associated gene-1) and BRM (Brahma). The transcriptional activator BRG1, also known as the ATP-dependent chromatin remodeling factor SMARCA4, is encoded by the SMARCA4 gene on chromosome 19. BRG1 is overexpressed in some cancerous tumors and is required for cancer cell proliferation. BRM, also known as the possible total transcriptional activator SNF2L2 and / or the ATP-dependent chromatin remodeling factor SMARCA2, is encoded by the SMARCA2 gene on chromosome 9 and has been shown to be essential for tumor cell growth in cells characterized by loss of BRG1 function mutations. Inactivation of BRG and / or BRM leads to downstream effects in the cell, including cell cycle arrest and tumor suppression.

[0004] Overview

[0005] The present invention is characterized by compounds that can be used to modulate the BAF complex. In some embodiments, said compounds can be used to treat conditions associated with alterations to the BAF complex, such as conditions associated with alterations to one or both of the BRG1 and BRM proteins. The compounds of the present invention, alone or in combination with other pharmaceutically active agents, can be used to treat such conditions.

[0006] In one aspect, the present invention provides compounds having the following structure:

[0007]

[0008] in

[0009] m can be 0, 1, 2, or 3;

[0010] n is 0, 1, 2, 3 or 4;

[0011] X 1 It can be -S-, -SO-, -SO2- or -S(O)(NH)-;

[0012] X 2 For N or CR 8 ;

[0013] R1 It is hydrogen or an optional substituted C1-C6 alkyl group;

[0014] Each R 2 and each R 3 Independently hydrogen, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl;

[0015] L 1 It is a substituted 9- to 10-membered bicyclic heterocyclic group or a substituted 9- to 10-membered bicyclic heteroaryl group;

[0016] L 2 C3-C does not exist and is an optional substitution. 10 cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted 5- to 14-membered heteroaryl or optionally substituted 4- to 14-membered heterocyclic group;

[0017] R 4 It is hydrogen, halogen, optionally substituted C1-C6 alkyl or optionally substituted C3-C 10 cycloalkyl;

[0018] R 5 The C1-C6 alkyl group, the C1-C6 heteroalkyl group, or the amino group may be optionally substituted, and R 6 It is hydrogen, halogen, cyano, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl or optionally substituted C3-C 10 cycloalkyl; or R 5 and R 6 Together with the attached atoms, they merge into optionally substituted 5- to 8-membered heterocyclic groups;

[0019] Each R 7 Independently, it is an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 heteroalkyl group, a halogen, or an optionally substituted C3-C... 10 cycloalkyl, optionally substituted C3-C 10 Cycloalkyl C1-C6 alkyl, optionally substituted 5- to 14-membered heteroaryl, optionally substituted 4- to 14-membered heterocyclic, -N(R 7A )2 or -OR 7A , where each R 7A Independently H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 cycloalkyl, optionally substituted C6-C 10 aryl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 4- to 14-membered heterocyclic, or two twin Rs 7AThe group, together with the attached atom, merges to form an optionally substituted 5- to 10-membered heteroaryl or an optionally substituted 4- to 10-membered heterocyclic group; or two twin R groups. 7 The group and the attached atom combine to form a carbonyl group;

[0020] R 8 It is hydrogen, halogen, optionally substituted C1-C6 alkyl or optionally substituted C3-C 10 cycloalkyl; and

[0021] R 9 It is hydrogen or halogen;

[0022] Or its pharmaceutically acceptable salt.

[0023] In some implementations, the variables of the compound of formula I are as follows:

[0024] m can be 0, 1, 2, or 3;

[0025] n is 0, 1, 2, 3 or 4;

[0026] X 1 It can be S, SO, SO2 or S(O)(NH);

[0027] X 2 For N or CR 8 ;

[0028] R 1 It is hydrogen or an optional substituted C1-C6 alkyl group;

[0029] Each R 2 and each R 3 Independently hydrogen, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl;

[0030] L 1 It is a substituted 9- to 10-membered bicyclic heterocyclic group or a substituted 9- to 10-membered bicyclic heteroaryl group;

[0031] L 2 C3-C does not exist and is an optional substitution. 10 cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted 5- to 10-membered heteroaryl or optionally substituted 4- to 10-membered heterocyclic;

[0032] R 4 It is hydrogen, halogen, optionally substituted C1-C6 alkyl or optionally substituted C3-C 10 cycloalkyl;

[0033] R 5The C1-C6 alkyl group, the C1-C6 heteroalkyl group, or the amino group may be optionally substituted, and R 6 It is hydrogen, halogen, cyano, optionally substituted C1-C6 alkyl or optionally substituted C3-C 10 cycloalkyl; or R 5 and R 6 Together with the attached atoms, they merge into optionally substituted 5- to 8-membered heterocyclic groups;

[0034] Each R 7 Independently, it is an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 heteroalkyl group, a halogen, or an optionally substituted C3-C... 10 Cycloalkyl, optionally substituted 5- to 10-membered heteroaryl, optionally substituted 4- to 10-membered heterocyclic, -N(R 7A )2 or -OR 7A , where each R 7A Independently H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 cycloalkyl, optionally substituted C6-C 10 aryl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 4- to 10-membered heterocyclic, or two twin Rs 7A The group and the attached atom are combined to form an optionally substituted 5- to 10-membered heteroaryl or an optionally substituted 4- to 10-membered heterocyclic group;

[0035] R 8 It is hydrogen, halogen, optionally substituted C1-C6 alkyl or optionally substituted C3-C 10 cycloalkyl; and

[0036] R 9 It is hydrogen;

[0037] Or its pharmaceutically acceptable salt.

[0038] In some implementations, L 2 C3-C does not exist and is an optional substitution. 10 cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 4- to 10-membered heterocyclic group.

[0039] In some implementations, each R 7 Independently, it is an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 heteroalkyl group, a halogen, or an optionally substituted C3-C... 10 Cycloalkyl, optionally substituted 5- to 10-membered heteroaryl, optionally substituted 4- to 10-membered heterocyclic, -N(R 7A )2 or -OR7A , where each R 7A Independently H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 cycloalkyl, optionally substituted C6-C 10 aryl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 4- to 10-membered heterocyclic, or two twin Rs 7A The group and the attached atom are combined to form an optionally substituted 5- to 10-membered heteroaryl or an optionally substituted 4- to 10-membered heterocyclic group.

[0040] In some implementation schemes, R 5 and R 6 Together with the attached atoms, they merge into optionally substituted 5- to 8-membered heterocyclic groups. In some embodiments, R 5 and R 6 It merges with the attached atom to form an optionally substituted 7-membered heterocyclic group.

[0041] In some implementation schemes, R 5 It is an optionally substituted C1-C6 alkyl group. In some embodiments, R 5 The amino group is optionally substituted. In some embodiments, R 6 It is an optionally substituted C1-C6 alkyl group. In some embodiments, R 6 It is a halogen.

[0042] In some implementation schemes, X 1 For SO2. In some implementations, X 2 For CR 8 .

[0043] In some implementation schemes,

[0044] Groups with the following structures in

[0045] Z represents CH2, CO, or C(R) X2 )2;

[0046] Each R X1 Independently, it is an optionally substituted C1-C6 alkyl or halogen, or two twinned Rs. X1 The group and the attached atom combine to form a carbonyl group;

[0047] Each R X2 Independently H or optionally substituted C1-C6 alkyl; and

[0048] p can be 0, 1, 2, 3, or 4.

[0049] In some implementation schemes, Groups with the following structures in

[0050] Z represents CH2, CO, or C(R) X2 )2;

[0051] Each R X1 Independently, it is an optionally substituted C1-C6 alkyl or halogen, or two twinned Rs. X1 The group and the attached atom combine to form a carbonyl group;

[0052] Each R X2 Independently H or optionally substituted C1-C6 alkyl; and

[0053] p can be 0, 1, 2, 3, or 4.

[0054] In some implementation schemes, Groups with the following structures in

[0055] Z represents CH2, CO, or C(R) X2 )2;

[0056] Each R X1 Independently, it is an optionally substituted C1-C6 alkyl or halogen, or two twinned Rs. X1 The group and the attached atom combine to form a carbonyl group;

[0057] Each R X2 Independently hydrogen or optionally substituted C1-C6 alkyl;

[0058] p is 0, 1, 2, 3, or 4; and

[0059] q is 0 or 1.

[0060] In some implementation schemes, Groups with the following structures in

[0061] Z represents CH2, CO, or C(R) X2 )2;

[0062] Each R X1 Independently, it is an optionally substituted C1-C6 alkyl or halogen, or two twinned Rs. X1 The group and the attached atom combine to form a carbonyl group;

[0063] Each R X2 Independently hydrogen or optionally substituted C1-C6 alkyl; and

[0064] p can be 0, 1, 2, 3, or 4.

[0065] In some implementation schemes, Groups with the following structures in

[0066] It can be a single bond or a double bond;

[0067] Each R X1 Independently, it is an optionally substituted C1-C6 alkyl or halogen, or two twinned Rs. X1 The group and the attached atom combine to form a carbonyl group;

[0068] R X2 It is hydrogen or optionally substituted C1-C6 alkyl; and

[0069] p can be 0, 1, 2, 3, or 4.

[0070] In some implementation schemes, Groups with the following structures in

[0071] Each R X1 Independently, it is an optionally substituted C1-C6 alkyl or halogen, or two twinned Rs. X1 The group and the attached atom combine to form a carbonyl group;

[0072] R X2 It is hydrogen or optionally substituted C1-C6 alkyl; and

[0073] p can be 0, 1, 2, 3, or 4.

[0074] In some implementation schemes, R 8 It is hydrogen.

[0075] In some implementation schemes, R 8 It is a halogen.

[0076] In some implementation schemes, R 8 It is an optional substituted C3-C8 cycloalkyl group.

[0077] In some implementation schemes, X 2 Let N be the number of elements in the array.

[0078] In some implementation schemes, Groups with the following structures in

[0079] Z represents CH2, CO, or C(R) X2 )2;

[0080] Each R X1 Independently, it is an optionally substituted C1-C6 alkyl or halogen, or two twinned Rs. X1 The group and the attached atom combine to form a carbonyl group;

[0081] Each R X2 It is independently hydrogen or optionally substituted C1-C6 alkyl; and

[0082] p can be 0, 1, 2, 3, or 4.

[0083] In some implementation schemes, Groups with the following structures in

[0084] Each R X1 Independently, it is an optionally substituted C1-C6 alkyl or halogen, or two twinned Rs. X1 The group and the attached atom combine to form a carbonyl group or a C3-C8 cycloalkyl ring, or two ortho-R groups. X1 The group and the attached atom also merge together to form a C3-C8 cyclic alkyl ring;

[0085] p is 0, 1, 2, 3, or 4; and

[0086] q can be 0, 1, or 2.

[0087] In some implementation schemes, Groups with the following structures in

[0088] Each R X1 Independently, it is an optionally substituted C1-C6 alkyl or halogen, or two twinned Rs. X1 The group and the attached atom combine to form a carbonyl group; and p is 0, 1, 2, 3 or 4.

[0089] In some implementations, at least one R X1 It is an optional substituted C1-C6 alkyl group.

[0090] In some implementations, -L 2 -(R 7 ) n Groups with the following structures:

[0091] In some implementations, at least one R X1 It is a halogen.

[0092] In some implementations, at least two twin R X1 The group and the atom it is attached to combine to form a carbonyl group.

[0093] In some implementations, L 1 It is a 9- to 10-membered bicyclic heteroaryl group with optional substitution.

[0094] In some implementations, L 1 for

[0095]

[0096] in

[0097] X 3 X 4 X 5 X 6 X 7 and X 8 Each is independently N or CR L1 ;

[0098] Each R L1 Independently, it is H, halogen, or optionally substituted C1-C6 alkyl;

[0099] A 1 Bond to –(C(R) 2 (R) 3 )) m -;and

[0100] A 2 Bond to L 2 .

[0101] In some implementations, L 1 for

[0102] In some implementations, L 1 for

[0103] In some implementations, L 1 for

[0104] In some implementations, L 1 for

[0105] In some implementations, L 1 for

[0106] In some implementations, L 1 for Where A 1 Bond to –(C(R) 2 (R) 3 )) m -; and A 2 Bond to L 2 .

[0107] In some implementations, L 2 It is a 5- to 10-membered heteroaryl group that is optionally substituted.

[0108] In some implementations, -L 2 -(R7 ) n Groups with the following structures:

[0109]

[0110]

[0111]

[0112] In some implementations, -L 2 -(R 7 ) n Groups with the following structures:

[0113]

[0114] In some implementations, -L 2 -(R 7 ) n Groups with the following structures:

[0115]

[0116] In some implementations, -L 2 -(R 7 ) n Groups with the following structures:

[0117] In some implementations, -L 2 -(R 7 ) n Groups with the following structures:

[0118] In some implementations, -L 2 -(R 7 ) n Groups with the following structures:

[0119] In some implementations, -L 2 -(R 7 ) n Groups with the following structures:

[0120] In some implementations, -L 2 -(R 7 ) n Groups with the following structures:

[0121] In some implementations, -L 2 -(R 7 ) nGroups with the following structures:

[0122] In some implementations, -L 2 -(R 7 ) n Groups with the following structures:

[0123] In some implementations, L 2 C6-C as an optional substitute 10 Aryl.

[0124] In some implementations, n is 1. In some implementations, n is 2. In some implementations, n is 3.

[0125] In some implementation schemes, R 7 It is an optionally substituted C1-C6 alkyl group. In some embodiments, R 7 It is an optionally substituted C1-C6 heteroalkyl group. In some embodiments, R 7 The substituted 4- to 14-membered heterocyclic group is used. In some embodiments, R 7 It is optionally substituted with an nitrogen-containing heterocyclic butyl group or optionally substituted with a morpholino group. In some embodiments, R 7 C3-C as an optional substitute 10 Cycloalkyl. In some embodiments, R 7 It is an optionally substituted cyclopropyl or optionally substituted cyclobutyl. In some embodiments, R 7 -N(R) 7A )2. In some implementation schemes, R 7 It is optionally substituted N-azacyclobutane or optionally substituted N-morpholino. In some embodiments, the two twin Rs 7 The group, together with the attached atom, merges into an optionally substituted 4- to 10-membered heterocyclic group. In some embodiments, at least one R 7 For -OR 7A In some implementations, R 7A C is an optional replacement 1-6 alkyl.

[0126] In some implementations, n is 0.

[0127] In some implementations, at least one R 7The compounds are cyclopropyl, 2,2-difluorocyclopropyl, difluoromethoxy, 2,6-dimethylmorpholin-4-yl, N-azacyclobutyl, 3-fluorocyclobutyl, 2-methoxyethyl, ethoxy, methoxy, 2,2-difluoroethoxy, 2,2-difluoroethyl, trifluoromethyl, isopropyl, methyl, acetyl, fluorine, chlorine, 1-methylpyrazol-3-yl, dimethylamino, N-methyl-N-(2-methoxyethyl)-amino, N-ethyl-N-(2-methoxyethyl)-amino, N-(2-propyl)-N-(2-methoxyethyl)-amino, 2-methoxyethylamino, 3-aza-8-oxa-bicyclo[4.3]. [0] Non-3-yl, 3-aza-7-oxa-bicyclo[4.3.0] Non-3-yl, 1-fluorocyclobut-1-yl, 3-fluoropyrrolidine-1-yl, 3-methoxypyrrolidine-1-yl, oxacyclobutane-3-yl, N-methyldihydroindole-4-yl, 2,2-difluoro-3-methylcycloprop-1-yl, 3-methoxyazacyclobutane-1-yl, 3-methoxypiperidin-1-yl, 1,2-dimethyl-7-azaindole-4-yl, 1-methyl-7-azaindole-4-yl, 2,3-methylenedioxyphenyl, N-methyl-N-(3-oxacyclobutyl)amino, 3-oxacyclobutyloxy, 1, 1-Difluoro-5-azaspiro[2.3]hex-5-yl, 1-fluoromethyl-cyclopropyl, N-(3-tetrahydrofuranyl)methylamino, N-dihydroindolyl, N-1,4-oxazaheptanylcycloyl, 2-fluoro-2-propyl, 1,1-difluoro-2-propyl, 2,2-difluoro-1-methylcyclopropyl-1-yl, 1-methylcyclopropyl, 4,4-difluoropiperidin-1-yl, 2-methoxyethoxy, 3,3-difluorocyclobut-1-yl, N-methyl-N-1-methoxypropyl-2-ylamino, 1-methoxypropyl-2-ylamino, 1-methoxyethyl, 4-methylpiperazinyl, 3-methylmorpholinyl, 2,2-difluoropropyl Oxygen, 3-methoxycyclobutyl, methylamino, 4-dimethylamino-3,3-difluoropiperidinyl, 4-methylamino-3,3-difluoropiperidinyl, 3,3-difluoropyrrolidinyl, N-methyl-N-3-methoxycyclobutylamino, 1-methylpyrazol-5-yl, 6-oxa-3-azabicyclo[3.1.1]hept-3-yl, cyclopropoxy, 2,6-dimethylpyridin-4-yl, 2-methylpyrrolidinyl, 4-oxabicyclo[4.1.0]hept-1-yl, N-methyl-N-(2,6-dimethyltetrahydropyran-4-yl)amino or N-methyl-N-3-methyloxacyclobutane-3-ylmethylamino.

[0128] In some implementation schemes, R 1 It is hydrogen.

[0129] In another aspect, the present invention provides compounds selected from compounds 1-308 in Table 1A and their pharmaceutically acceptable salts.

[0130] Table 1A. Compounds of the present invention

[0131]

[0132]

[0133]

[0134]

[0135]

[0136]

[0137]

[0138]

[0139]

[0140]

[0141]

[0142]

[0143]

[0144]

[0145]

[0146]

[0147]

[0148]

[0149]

[0150]

[0151]

[0152]

[0153]

[0154]

[0155]

[0156]

[0157]

[0158]

[0159]

[0160]

[0161]

[0162]

[0163]

[0164]

[0165]

[0166]

[0167]

[0168]

[0169]

[0170]

[0171]

[0172]

[0173]

[0174]

[0175]

[0176]

[0177]

[0178]

[0179]

[0180]

[0181]

[0182]

[0183]

[0184]

[0185]

[0186]

[0187]

[0188]

[0189]

[0190] On the other hand, the present invention provides compounds selected from compounds 309-856 in Table 1B and their pharmaceutically acceptable salts.

[0191] Table 1B. Compounds of the present invention (cyclic and non-cyclic sulfones)

[0192]

[0193]

[0194]

[0195]

[0196]

[0197]

[0198]

[0199]

[0200]

[0201]

[0202]

[0203]

[0204]

[0205]

[0206]

[0207]

[0208]

[0209]

[0210]

[0211]

[0212]

[0213]

[0214]

[0215]

[0216]

[0217]

[0218]

[0219]

[0220]

[0221]

[0222]

[0223]

[0224]

[0225]

[0226]

[0227]

[0228]

[0229]

[0230]

[0231]

[0232]

[0233]

[0234]

[0235]

[0236]

[0237]

[0238]

[0239]

[0240]

[0241]

[0242]

[0243]

[0244]

[0245]

[0246]

[0247]

[0248]

[0249]

[0250]

[0251]

[0252]

[0253]

[0254]

[0255]

[0256]

[0257]

[0258]

[0259]

[0260]

[0261]

[0262]

[0263]

[0264]

[0265]

[0266]

[0267]

[0268]

[0269] In some embodiments, the compound has at least 5 BRG1 ICs. 50 With BRM IC 50 The ratio. In some embodiments, the compound has BRG1 IC 50 With BRM IC 50 The ratio is at least 7. In some embodiments, the compound has at least 10 BRG1 ICs. 50 With BRM IC 50 The ratio. In some embodiments, the compound has a BRG1 IC ratio of at least 15. 50 With BRM IC 50 The ratio. In some embodiments, the compound has at least 20 BRG1 IC. 50 With BRM IC 50 The ratio. In some embodiments, the compound has a BRG1 IC ratio of at least 25. 50 With BRMIC 50 The ratio. In some embodiments, the compound has a BRG1IC ratio of at least 30. 50 With BRM IC 50 The ratio.

[0270] In another aspect, the present invention is characterized by a pharmaceutical composition comprising any of the above-described compounds and a pharmaceutically acceptable excipient.

[0271] In another aspect, the present invention is characterized by a method for reducing the activity of the BAF complex in cells, the method comprising exposing the cells to an effective amount of any of the above-described compounds or pharmaceutical compositions thereof.

[0272] In some implementations, the cells are cancer cells.

[0273] In another aspect, the present invention is characterized by a method for treating BAF complex-related conditions in a subject in need, the method comprising administering to the subject an effective amount of any of the above-mentioned compounds or a pharmaceutical composition thereof.

[0274] In some implementations, the BAF complex-related condition is cancer.

[0275] In another aspect, the present invention is characterized by a method for inhibiting BRM, the method comprising exposing cells to an effective amount of any of the above-described compounds or pharmaceutical compositions thereof. In some embodiments, the cells are cancer cells.

[0276] In another aspect, the present invention is characterized by a method for inhibiting BRG1, the method comprising exposing the cells to an effective amount of any of the above-described compounds or pharmaceutical compositions thereof. In some embodiments, the cells are cancer cells.

[0277] In another aspect, the present invention is characterized by a method for inhibiting BRM and BRG1, the method comprising exposing cells to an effective amount of any of the above-described compounds or pharmaceutical compositions thereof. In some embodiments, the cells are cancer cells.

[0278] In another aspect, the present invention is characterized by a method for treating a condition associated with a loss-of-function mutation of BRG1 in a subject in need, the method comprising administering to the subject an effective amount of any of the above-mentioned compounds or a pharmaceutical composition thereof.

[0279] In some implementations, the condition associated with the BRG1 loss-of-function mutation is cancer. In other implementations, the subject is identified as having a BRG1 loss-of-function condition, such as being identified as having BRG1 loss-of-function cancer (e.g., the cancer has been identified as including cancer cells with BRG1 loss-of-function).

[0280] In another aspect, the present invention is characterized by a method for inducing apoptosis in cells, the method comprising contacting the cells with an effective amount of any of the above-described compounds or pharmaceutical compositions thereof. In some embodiments, the cells are cancer cells.

[0281] In another aspect, a method for treating cancer in subjects in need, the method comprising administering to the subject an effective amount of any of the above-mentioned compounds or a pharmaceutical composition thereof.

[0282] In some embodiments of any of the above methods, the cancer is non-small cell lung cancer, colorectal cancer, bladder cancer, primary cancer of unknown cause, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, esophageal and gastric cancer, pancreatic cancer, hepatobiliary cancer, soft tissue sarcoma, ovarian cancer, head and neck cancer, renal cell carcinoma, bone cancer, non-Hodgkin's lymphoma, small cell lung cancer, prostate cancer, embryonal tumor, germ cell tumor, cervical cancer, thyroid cancer, salivary gland cancer, gastrointestinal neuroendocrine tumor, uterine sarcoma, gastrointestinal stromal tumor, CNS cancer, thymic tumor, adrenocortical carcinoma, appendix cancer, small bowel cancer, or penile cancer.

[0283] In some embodiments of any of the above methods, the cancer is non-small cell lung cancer, colorectal cancer, bladder cancer, primary cancer of unknown cause, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, or penile cancer.

[0284] In some embodiments of any of the above methods, the cancer is drug-resistant cancer or cancer unresponsive to prior therapies (e.g., vemurafenib, dacarbazine, CTLA4 inhibitors, PD1 inhibitors, interferon therapy, BRAF inhibitors, MEK inhibitors, radiotherapy, temozolomide, irinotecan, CAR-T therapy, Herceptin, perjeta, tamoxifen, xeloda, docetaxel, platinum-based agents such as carboplatin, taxanes such as paclitaxel and docetaxel, ALK inhibitors, MET inhibitors, alimta, abraxane for injection). It responds to gemcitabine, avastin, halaven mesylate, neratinib, PARP inhibitors, ARN810, mTOR inhibitors, topotecan, gemzar, VERT inhibitors, folate receptor antagonists, demcizumab, fosbretabulin, or PDL1 inhibitors.

[0285] In some embodiments of any of the foregoing methods, the cancer has been or has been identified as having a BRG1 mutation. In some embodiments of any of the foregoing methods, the BRG1 mutation is homozygous. In some embodiments of any of the foregoing methods, the cancer does not have or has been identified as not having an epidermal growth factor receptor (EGFR) mutation. In some embodiments of any of the foregoing methods, the cancer does not have or has been identified as not having an anaplastic lymphoma kinase (ALK) driver mutation. In some embodiments of any of the foregoing methods, the cancer has or has been identified as having a KRAS mutation. In some embodiments of any of the foregoing methods, the BRG1 mutation is in the ATPase catalytic domain of the protein. In some embodiments of any of the foregoing methods, the BRG1 mutation is a deletion at the C-terminus of BRG1.

[0286] In another aspect, this disclosure provides a method for treating BAF-related conditions (e.g., cancer or viral infection) in subjects in need. The method comprises contacting cells with an effective amount of any of the above-described compounds or a pharmaceutically acceptable salt thereof, or any of the above-described pharmaceutical compositions. In some embodiments, the symptom is a viral infection, specifically a virus infection of the Retroviridae family, such as lentiviruses (e.g., human immunodeficiency virus (HIV) and delta retroviruses (e.g., human T-cell leukemia virus I (HTLV-I), human T-cell leukemia virus II (HTLV-II)), Hepadnaviridae family (e.g., hepatitis B virus (HBV)), Flaviviridae family (e.g., hepatitis C virus (HCV)), Adenoviridae family (e.g., human adenovirus), Herpesviridae family (e.g., human cytomegalovirus (HCMV), Epstein-Barr virus, herpes simplex virus 1 (HSV-1), herpes simplex virus 2 (HSV-2), human herpesvirus 6 (HHV-6), herpesvirus K*, CMV, varicella-zoster virus (VZV). The diseases described include those belonging to the Papillomaviridae family (e.g., human papillomavirus (HPV, HPVE1)), Parvoviridae family (e.g., parvovirus B19), Polyomaviridae family (e.g., JC virus and BK virus), Paramyxoviridae family (e.g., measles virus), and Papillomaviridae family (e.g., rubella virus). In some embodiments, the disease is Coffin Siris, neurofibromatosis (e.g., NF-1, NF-2, or Schwannomatosis), or multiple meningioma.

[0287] In another aspect, this disclosure provides a method for treating a viral infection in a subject in need. The method comprises administering to the subject an effective amount of any of the aforementioned compounds or a pharmaceutically acceptable salt thereof, or any of the aforementioned pharmaceutical compositions. In some embodiments, the viral infection is a virus infection of the Retroviridae family, such as lentiviruses (e.g., human immunodeficiency virus (HIV) and delta retroviruses (e.g., human T-cell leukemia virus I (HTLV-I), human T-cell leukemia virus II (HTLV-II)), hepatoviridae (e.g., hepatitis B virus (HBV)), flaviviridae (e.g., hepatitis C virus (HCV)), adenoviridae (e.g., human adenovirus), herpesviruses (e.g., human cytomegalovirus (HCMV), Epstein-Barr virus, herpesvirus), herpes simplex virus 1 (HSV-1), herpes simplex virus 2 (HSV-2), human herpesvirus 6 (HHV-6), herpesvirus K*, CMV, varicella-zoster virus), and papillomaviruses (e.g., human papillomavirus (HPV, HPV...). E1), Parvoviridae (e.g., parvovirus B19), Polyomaviridae (e.g., JC virus and BK virus), Paramyxoviridae (e.g., measles virus), or Clonorchiidae (e.g., rubella virus).

[0288] In another aspect, the present invention is characterized by a method for treating a subject in need of melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or blood cancer, the method comprising administering to the subject an effective amount of any of the above-mentioned compounds or pharmaceutical compositions thereof.

[0289] In another aspect, the present invention is characterized by a method for reducing tumor growth of melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or blood cancer in a subject in need, said method comprising administering to said subject an effective amount of any of the above-mentioned compounds or a pharmaceutical composition thereof.

[0290] In another aspect, the present invention is characterized by a method for inhibiting metastatic progression of melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or blood cancer in a subject, the method comprising administering an effective amount of any of the above-mentioned compounds or a pharmaceutical composition thereof.

[0291] In another aspect, the present invention is characterized by a method for inhibiting metastatic colonization of melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or blood cancer in a subject, the method comprising administering an effective amount of any of the above-mentioned compounds or a pharmaceutical composition thereof.

[0292] In another aspect, the present invention is characterized by a method for reducing the levels and / or activity of BRG1 and / or BRM in melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or hematologic malignancies, the method comprising exposing cells to an effective amount of any of the above-mentioned compounds or pharmaceutical compositions thereof.

[0293] In some implementations of any of the above aspects, melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or blood cancer cells are present in the subject.

[0294] In some embodiments of any of the foregoing aspects, the effective amount of the compound reduces the level and / or activity of BRG1 by at least 5% compared to the reference (e.g., 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%). In some embodiments, the effective amount of the compound reduces the level and / or activity of BRG1 by at least 50% compared to the reference (e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%). In some embodiments, the effective amount of the compound reduces the level and / or activity of BRG1 by at least 90% (e.g., 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%).

[0295] In some embodiments, the effective amount of the compound reduces the level and / or activity of BRG1 by at least 5% (e.g., 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) compared to the reference, for at least 12 hours (e.g., 14 hours, 16 hours, 18 hours, 20 hours, 22 hours, 24 hours, 30 hours, 36 hours, 48 ​​hours, 72 hours, or more). In some embodiments, the effective amount of the compound reduces the level and / or activity of BRG1 by at least 5% (e.g., 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) compared to the reference, for at least 4 days (e.g., 5 days, 6 days, 7 days, 14 days, 28 days, or more).

[0296] In some embodiments of any of the foregoing aspects, the effective amount of the compound reduces the level and / or activity of BRM by at least 5% compared to the reference (e.g., 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%). In some embodiments, the effective amount of the compound reduces the level and / or activity of BRM by at least 50% compared to the reference (e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%). In some embodiments, the effective amount of the compound reduces the level and / or activity of BRM by at least 90% (e.g., 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%).

[0297] In some embodiments, the effective amount of the compound reduces the level and / or activity of the BRM by at least 5% (e.g., 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) compared to the reference, for at least 12 hours (e.g., 14 hours, 16 hours, 18 hours, 20 hours, 22 hours, 24 hours, 30 hours, 36 hours, 48 ​​hours, 72 hours, or more). In some embodiments, the effective amount of the compound reduces the level and / or activity of the BRM by at least 5% (e.g., 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) compared to the reference, for at least 4 days (e.g., 5 days, 6 days, 7 days, 14 days, 28 days, or more).

[0298] In some embodiments, the subject has cancer. In some embodiments, the cancer expresses BRG1 and / or BRM proteins and / or the cells or subject have been identified as expressing BRG1 and / or BRM. In some embodiments, the cancer expresses BRG1 protein and / or the cells or subject have been identified as expressing BRG1. In some embodiments, the cancer expresses BRM protein and / or the cells or subject have been identified as expressing BRM. In some embodiments, the cancer is melanoma (e.g., ocular lentigines melanoma, mucosal melanoma, or cutaneous melanoma). In some embodiments, the cancer is prostate cancer. In some embodiments, the cancer is a hematologic malignancy, such as multiple myeloma, large cell lymphoma, acute T-cell leukemia, acute myeloid leukemia, myelodysplastic syndrome, immunoglobulin Aλ myeloma, diffuse mixed histiocytic and lymphocytic lymphoma, B-cell lymphoma, acute lymphoblastic leukemia (e.g., T-cell acute lymphoblastic leukemia or B-cell acute lymphoblastic leukemia), diffuse large cell lymphoma, or non-Hodgkin's lymphoma. In some embodiments, the cancer is breast cancer (e.g., ER-positive breast cancer, ER-negative breast cancer, triple-positive breast cancer, or triple-negative breast cancer). In some embodiments, the cancer is bone cancer (e.g., Ewing sarcoma). In some embodiments, the cancer is renal cell carcinoma (e.g., microphthalmia transcription factor (MITF) family translocation renal cell carcinoma (tRCC)). In some embodiments, the cancer is metastatic (e.g., the cancer has spread to the liver). Metastatic cancer may include cells exhibiting migration and / or invasion of migratory cells and / or cells exhibiting endothelial recruitment and / or angiogenesis. In other embodiments, the metastatic cancer is a cell-migrating cancer. In other embodiments, the cell-migrating cancer is a non-metastatic cell-migrating cancer. The metastatic cancer may be a cancer that has spread by inoculation of the surface of the peritoneum, pleura, pericardium, or subarachnoid space. Alternatively, the metastatic cancer may be a cancer that has spread via the lymphatic system or a hematogenous cancer. In some embodiments, the effective amount of the compound of the invention is an amount that effectively inhibits the metastatic colonization of the cancer to the liver.

[0299] In some embodiments, the cancer carries a mutation in GNAQ. In some embodiments, the cancer carries a mutation in GNA11. In some embodiments, the cancer carries a mutation in PLCB4. In some embodiments, the cancer carries a mutation in CYSLTR2. In some embodiments, the cancer carries a mutation in BAP1. In some embodiments, the cancer carries a mutation in SF3B1. In some embodiments, the cancer carries a mutation in EIF1AX. In some embodiments, the cancer carries a TFE3 translocation. In some embodiments, the cancer carries a TFEB translocation. In some embodiments, the cancer carries a MITF translocation. In some embodiments, the cancer carries an EZH2 mutation. In some embodiments, the cancer carries a SUZ12 mutation. In some embodiments, the cancer carries an EED mutation.

[0300] In some embodiments of any of the above methods, the method further includes administering an anticancer therapy to the subject or exposing cells to an anticancer therapy, such as a chemotherapeutic agent or cytotoxic agent, immunotherapy, surgery, radiotherapy, thermotherapy, or photocoagulation therapy, or combinations thereof. In some embodiments, the anticancer therapy is a chemotherapeutic agent or cytotoxic agent, such as antimetabolites, antimitotic drugs, antitumor antibiotics, asparagine-specific enzymes, bisphosphonates, antitumor drugs, alkylating agents, DNA repair enzyme inhibitors, histone deacetylase inhibitors, corticosteroids, demethylating agents, immunomodulators, Janus-associated kinase inhibitors, phosphinositide 3-kinase inhibitors, proteasome inhibitors, or tyrosine kinase inhibitors, or combinations thereof.

[0301] In some embodiments of any of the above methods, the compounds of the present invention are used in combination with another anticancer therapy for treating ocular pigmentary melanoma, such as surgery, MEK inhibitors, and / or PKC inhibitors. For example, in some embodiments, the method further includes performing surgery before, after, or simultaneously with the administration of the compounds of the present invention. In some embodiments, the method further includes administering MEK inhibitors and / or PKC inhibitors before, after, or simultaneously with the administration of the compounds of the present invention.

[0302] In some embodiments, the anticancer therapy and the compound of the present invention are administered to each other within 28 days, and each is administered in an amount that is effective in treating the subject together.

[0303] In some implementations, the subject or cancer has and / or has been identified as having a BRG1 loss-of-function mutation.

[0304] In some embodiments, the cancer is resistant to one or more chemotherapeutic agents or cytotoxic agents (e.g., the cancer has been identified as resistant to chemotherapeutic agents or cytotoxic agents by genetic markers, or may be resistant to chemotherapeutic agents or cytotoxic agents, such as cancers that do not respond to chemotherapeutic agents or cytotoxic agents). In some embodiments, the cancer does not respond to one or more chemotherapeutic agents or cytotoxic agents. In some embodiments, the cancer is resistant to or unresponsive to dacarbazine, temozolomide, cisplatin, treosulfan, formustin, IMCgp100, CTLA-4 inhibitors (e.g., ipilimumab), PD-1 inhibitors (e.g., nivolumab or pembrolizumab), PD-L1 inhibitors (e.g., atezolizumab, avelumab, or durvalumab), mitogen-activated protein kinase (MEK) inhibitors (e.g., selumetinib, binimetinib, or tametinib), and / or protein kinase C (PKC) inhibitors (e.g., sotrastaurin or IDE196).

[0305] In some embodiments, the cancer is resistant to or unresponsive to previously administered therapeutic agents for treating ocular retinal melanoma, such as MEK inhibitors or PKC inhibitors. For example, in some embodiments, the cancer is resistant to or unresponsive to mitogen-activated protein kinase (MEK) inhibitors (e.g., selmetinib, bimetinib, or trametinib) and / or protein kinase C (PKC) inhibitors (e.g., sotrastuxin or IDE196).

[0306] Chemical terminology

[0307] The terminology used herein is for the purpose of describing a particular implementation and is not intended to be restrictive.

[0308] For any of the following chemical definitions, the number following the atomic symbol indicates the total number of atoms of that element present in a particular chemical moiety. As will be understood, other atoms, such as H atoms or substituents, may be present as required to satisfy the valence of the atoms. For example, an unsubstituted C2 alkyl group has the formula –CH2CH3. When used with groups as defined herein, references to the number of carbon atoms include divalent carbons in acetals and ketals, but exclude carbonyl carbons in acyl, ester, carbonate, or carbamate groups. References to the number of oxygen, nitrogen, or sulfur atoms in heteroaryl groups include only those atoms that form part of the heterocycle.

[0309] As used herein, the term "acyl" means an H or alkyl group connected to a parent molecule group via a carbonyl group as defined herein, and examples include formyl (i.e., carboxylaldehyde), acetyl, trifluoroacetyl, propionyl, and butyryl. Exemplary unsubstituted acyl groups include 1 to 6, 1 to 11, or 1 to 21 carbons.

[0310] As used herein, the term "alkenyl" refers to a branched or straight-chain monovalent saturated aliphatic hydrocarbon group (e.g., 2 to 16 carbon atoms, 2 to 10 carbon atoms, or 2 to 6 carbon atoms) having 2 to 20 carbon atoms. Alkenyl groups can be, for example, monovalent or polyvalent. Those skilled in the art will recognize from the context the number of available valences.

[0311] As used herein, the term "alkyl" refers to a branched or straight-chain monovalent saturated aliphatic hydrocarbon group having 1-20 carbon atoms (e.g., 1-16 carbon atoms, 1-10 carbon atoms, 1-6 carbon atoms, or 1-3 carbon atoms). Alkyl groups can be, for example, monovalent or polyvalent. Those skilled in the art will recognize the number of available valences from the context.

[0312] As used herein, the term "amino" represents –N(R N1 )2, where each R N1 Independently, H, OH, NO2, N(R) N2 2. SO2OR N2 SO2R N2 SOR N2 N-protecting group, alkyl, alkoxy, aryl, aralkyl, cycloalkyl, acyl (e.g., acetyl, trifluoroacetyl or other groups described herein), heteroaryl or heterocyclic group, wherein each of these listed R N1 The group can be optionally substituted; or two R groups can be substituted. N1 They combine with the atoms they are attached to to form heterocyclic groups or heteroaryl groups, and each R in these groups... N2 Independently, it is H, alkyl, or aryl. The amino group of the present invention can be an unsubstituted amino group (i.e., –NH2) or a substituted amino group (i.e., –N(R)). N1 )2).

[0313] As used herein, the term "aryl" refers to an aromatic monocyclic or polycyclic aromatic group having 6 to 12 carbon atoms having at least one aromatic ring. Examples of such groups include, but are not limited to, phenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, 1,2-dihydronaphthyl, indenyl, and 1H-indenyl. Aryl groups can be, for example, monovalent or polyvalent. Those skilled in the art will recognize the number of available valences from the context.

[0314] As used herein, the term "arylalkyl" refers to an alkyl group that has been substituted with an aryl group. Exemplary unsubstituted arylalkyl groups have 7 to 30 carbons (e.g., 7 to 16 or 7 to 20 carbons, such as C1-C6 alkyl C6-C). 10 Aryl, C1-C 10 Alkyl C6-C 10 Aryl or C1-C 20 Alkyl C6-C 10 Aryl groups, such as benzyl and phenethyl. In some embodiments, the alkyl and aryl groups may each be further substituted with one, two, three, or four substituents as defined herein for the respective groups.

[0315] As used in this article, the term "azido" refers to the –N3 group.

[0316] As used herein, the term “bridged polycyclic alkyl” refers to a bridging polycyclic group of 5 to 20 carbons containing 1 to 3 bridges.

[0317] As used in this article, the term "cyano" refers to the –CN group.

[0318] As used in this article, the term "carbocyclic group" refers to a non-aromatic C3-C group. 12 Monocyclic, bicyclic, or tricyclic structures, wherein the rings are formed by carbon atoms. Carbocyclic structures include cycloalkyl and unsaturated carbocyclic groups.

[0319] As used herein, the term "cycloalkyl" refers to a saturated, non-aromatic, and monovalent single- or multi-carbon cyclic group having 3 to 10, preferably 3 to 6, carbon atoms. Further examples of this term include groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, and adamantyl. Cycloalkyl groups can be, for example, monovalent or polyvalent. Those skilled in the art will recognize the number of available valences from the context.

[0320] As used herein, the term "halogen" refers to a fluorine (fluorinated), chlorine (chlorinated), bromine (brominated), or iodine (iodinated) group.

[0321] As used herein, the term "heteroalkyl" refers to an alkyl group as defined herein, wherein one or more constituent carbon atoms have been replaced by nitrogen, oxygen, or sulfur. In some embodiments, the heteroalkyl group may be further substituted with one, two, three, or four substituents as described herein for alkyl groups. An example of a heteroalkyl group is "alkoxy," which, as used herein, refers to an alkyl-O- (e.g., methoxy and ethoxy). Heteroalkyl groups can be, for example, monovalent or polyvalent. Those skilled in the art will recognize from the context the number of available valences.

[0322] As used herein, the term "heteroaryl" refers to a monocyclic or polycyclic group of 5 to 14 atoms (e.g., 5 to 12 or 5 to 10) having at least one aromatic ring and comprising 1, 2, or 3 ring atoms selected from nitrogen, oxygen, and sulfur, with the remaining ring atoms being carbon. In some embodiments, the heteroaryl is a C1-C9 heteroaryl (e.g., a C2-C9 heteroaryl). One or two ring carbon atoms of the heteroaryl may be substituted with a carbonyl group. Examples of heteroaryls are pyridyl, pyrazolyl, benzoxazolyl, benzimidazolyl, benzothiazolyl, imidazolyl, oxazolyl, thiazolyl, benzimorpholinyl, benzopiperidinyl, and dihydroindolyl. Heteroaryls can be, for example, monovalent or polyvalent. Those skilled in the art will recognize the number of available valences from the context.

[0323] As used herein, the term "heteroaryl" refers to an alkyl group substituted with a heteroaryl group. Exemplary unsubstituted heteroaryl groups have 7 to 30 carbons (e.g., 7 to 16 or 7 to 20 carbons, such as C2-C9 heteroaryl C1-C6 alkyl, C2-C9 heteroaryl C1-C6 alkyl, etc.). 10 Alkyl or C2-C9 heteroaryl C1-C 20 Alkyl group). In some embodiments, the alkyl and heteroaryl groups may each be further substituted with one, two, three, or four substituents as defined herein for the respective groups.

[0324] As used herein, the term "heterocyclic group" refers to a monocyclic or polycyclic group having 3 to 14 (e.g., 4 to 12) atoms, having at least one ring comprising 1, 2, 3, or 4 ring atoms selected from N, O, or S, wherein none of the rings is aromatic. In some embodiments, the heterocyclic group is a C2-C9 heterocyclic group. Examples of heterocyclic groups include, but are not limited to, morpholino, thiomorpholino, piperazine, piperidinyl, pyrano, pyrrolidinyl, tetrahydropyrano, tetrahydrofurano, 1,3-dioxane, aziroxybicyclo[4.3.0]nonyl, and aziroxybicyclo[4.4.0]decyl. Heterocyclic groups can be, for example, monovalent or polyvalent. Those skilled in the art will recognize the number of valences available from the context.

[0325] As used herein, the term "heterocyclic alkyl" refers to an alkyl group substituted with a heterocyclic group. Exemplary unsubstituted heterocyclic alkyl groups have 7 to 30 carbons (e.g., 7 to 16 or 7 to 20 carbons, such as C2-C9 heterocyclic C1-C6 alkyl, C2-C9 heterocyclic C1-C6 alkyl, etc.). 10 Alkyl or C2-C9 heterocyclic C1-C 20 Alkyl group). In some embodiments, the alkyl group and the heterocyclic group may each be further substituted with one, two, three or four substituents as defined herein for the respective group.

[0326] As used herein, the term "hydroxyalkyl" refers to an alkyl group in which the alkyl group is replaced by a –OH group.

[0327] As used in this article, the term "hydroxyl group" refers to the –OH group.

[0328] As used herein, the term “N-protecting group” refers to those groups intended to protect amino groups from undesirable reactions during synthetic processes. Commonly used N-protecting groups are disclosed in Greene, “Protective Groups in Organic Synthesis,” 3rd edition (John Wiley & Sons, New York, 1999). N-Protecting groups include, but are not limited to, acyl, aromatic, or carbamoyl groups, such as formyl, acetyl, propionyl, neopentyl, tert-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthaloyl, o-nitrophenoxyacetyl, α-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, 4-nitrobenzoyl, and chiral auxiliaries, such as protected or unprotected D,L, or D,L-amino acids, such as alanine, leucine, and phenylalanine; sulfonyl groups, such as benzylsulfonyl and p-toluenesulfonyl; carbamate-forming groups, such as benzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 3,5-dimethoxybenzyl Oxycarbonyl, 2,4-20-dimethoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl, 3,4,5-trimethoxybenzyloxycarbonyl, 1-(p-biphenyl)-1-methylethoxycarbonyl, α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, diphenylmethoxycarbonyl, tert-butoxycarbonyl, diisopropylmethoxycarbonyl, isopropoxycarbonyl, ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl, 2,2,2,-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl, cyclopentoxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl and phenylthiocarbonyl, arylalkyl such as benzyl, triphenylmethyl and benzyloxymethyl, and silyl such as trimethylsilyl. Preferred N-protecting groups are alloc, formyl, acetyl, benzoyl, neopentyl, tert-butylacetyl, alanyl, benzyl, tert-butoxycarbonyl (Boc), and benzyloxycarbonyl (Cbz).

[0329] As used in this article, the term "nitro" refers to a –NO2 group.

[0330] As used in this article, the term "hydrothioyl" refers to the –SH group.

[0331] The alkyl, heteroalkyl, carbocyclic (e.g., cycloalkyl), aryl, heteroaryl, and heterocyclic groups may be substituted or unsubstituted. When substituted, there are typically 1 to 4 substituents unless otherwise stated. Substituents include, for example: alkyl (e.g., unsubstituted and substituted, wherein the substituents include any group described herein, such as aryl, halogen, hydroxyl), aryl (e.g., substituted and unsubstituted phenyl), carbocyclic (e.g., substituted and unsubstituted cycloalkyl), halogen (e.g., fluorine), hydroxyl, heteroalkyl (e.g., substituted and unsubstituted methoxy, ethoxy, or thioalkoxy), heteroaryl, heterocyclic, amino (e.g., NH2 or monoalkylamino or dialkylamino), azide, cyano, nitro, or thiohydrosulfonyl. Aryl, carbocyclic (e.g., cycloalkyl), heteroaryl, and heterocyclic groups may also be substituted with alkyl (unsubstituted and substituted, e.g., arylalkyl (e.g., substituted and unsubstituted benzyl)).

[0332] The compounds of this invention may have one or more asymmetric carbon atoms and may exist as optically pure enantiomers, mixtures of enantiomers such as racemates, optically pure diastereomers, mixtures of diastereomers, diastereomeric racemates, or mixtures of diastereomeric racemates. Optically active forms may be obtained, for example, by resolving racemates, by asymmetric synthesis, or by asymmetric chromatography (chromatography using chiral adsorbents or eluents). That is, some of the disclosed compounds may exist in a variety of stereoisomeric forms. Stereoisomers are compounds that differ only in their spatial arrangement. Enantiomers are mirror-image, non-overlapping stereoisomer pairs, most commonly because they contain an asymmetrically substituted carbon atom as a chiral center. "Enantiomer" means one of a pair of molecules that are mirror images of each other and non-overlapping. Diastereomers are stereoisomers that are not mirror-image related, most commonly because they contain two or more asymmetrically substituted carbon atoms and represent configurations of substituents surrounding one or more chiral carbon atoms. Enantiomers of compounds can be prepared, for example, by separating enantiomers from racemates using one or more well-known techniques and methods, such as chiral chromatography and separation methods based thereon. Those skilled in the art can readily determine suitable techniques and / or methods for separating enantiomers of the compounds described herein from racemic mixtures. "Racemate" or "racemic mixture" refers to a compound containing two enantiomers, wherein such mixtures do not exhibit optical activity; i.e., they do not rotate the plane of polarized light. "Geometric isomer" refers to isomers in which the substituent atoms are oriented differently relative to a carbon-carbon double bond, a cycloalkyl ring, or a bridged bicyclic system. Atoms on each side of the carbon-carbon double bond (except H) can be in E (substituents on opposite sides of the carbon-carbon double bond) or Z (substituents oriented on the same side) configurations. "R", "S", "S*", "R*", "E", "Z", "cis", and "trans" indicate configurations relative to the core molecule. Some disclosed compounds may exist as trans-blocked isomers. Rotation-restricted isomers are stereoisomers resulting from hindered rotation around a single bond, wherein the steric strain barrier of rotation is sufficiently high to allow for the separation of conformational isomers. The compounds of the present invention can be prepared as individual isomers through isomer-specific synthesis or by resolution from mixtures of isomers. Conventional resolution techniques include using an optically active acid to form a salt of the free base of each isomer in an isomer pair (followed by fractional crystallization and regeneration of the free base), using an optically active amine to form a salt of the acid form of each isomer in an isomer pair (followed by fractional crystallization and regeneration of the free acid), using an optically pure acid, amine, or alcohol to form an ester or amide of each isomer in an isomer pair (followed by chromatographic separation and removal of chiral auxiliaries), or using various well-known chromatographic methods to resolve mixtures of isomers of starting materials or final products.When the stereochemistry of a disclosed compound is named or described by structure, the named or described stereoisomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by weight relative to other stereoisomers. When a single enantiomer is named or described by structure, the described or named enantiomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by weight optical purity. When a single diastereomer is named or described by structure, the described or named diastereomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by weight purity. The percentage of optical purity is the ratio of the weight of the enantiomer to the weight of the enantiomer plus the weight of its optical isomer. The diastereomer purity by weight is the ratio of the weight of one diastereomer to the weight of all diastereomers. When the stereochemistry of a disclosed compound is named or described by structure, the named or described stereoisomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% pure in mole fraction relative to other stereoisomers. When a single enantiomer is named or described by structure, the described or named enantiomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% pure in mole fraction. When a single diastereomer is named or described by structure, the described or named diastereomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% pure in mole fraction. Purity percentage in mole fraction is the ratio of the number of moles of the enantiomer to the sum of the number of moles of its optical isomer. Similarly, mole fraction purity percentage is the ratio of the number of moles of the diastereomer to the sum of the number of moles of its isomer. When a disclosed compound is named or described structurally without indicating its stereochemistry and has at least one chiral center, it should be understood that the name or structure covers enantiomers of the compound that do not contain the corresponding optical isomer, racemic mixtures of the compound, or mixtures rich in one enantiomer relative to their corresponding optical isomers. When a disclosed compound is named or described structurally without indicating its stereochemistry and has two or more chiral centers, it should be understood that the name or structure covers diastereomers that do not contain other diastereomers, a number of diastereomers that do not contain other diastereomer pairs, mixtures of diastereomers, mixtures of diastereomer pairs, mixtures of diastereomers in which one diastereomer is enriched relative to other diastereomers, or mixtures of one or more diastereomers enriched relative to other diastereomers. The invention includes all of these forms.

[0333] The compounds disclosed herein also include all isotopes of the atoms present in the intermediates or final compounds. "Isotope" refers to an atom having the same atomic number but different mass numbers produced by different numbers of neutrons in its nucleus. For example, isotopes of hydrogen include tritium and deuterium.

[0334] Unless otherwise stated, the structures described herein are also intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. Exemplary isotopes that can be incorporated into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, for example... 2 H, 3 H, 11 C 13 C 14 C 13 N、 15 N、 15 O、 17 O、 18 O、 32 P, 33 P, 35 S, 18 F, 36 Cl、 123 I and 125 I. Isotope-labeled compounds (e.g.) 3 H and 14 Those labeled with C can be used for the determination of the tissue distribution of compounds or substrates. Tritium (i.e., 3 H) and carbon-14 (i.e. 14 C) Isotopes can be useful due to their ease of preparation and detectability. Furthermore, replacing, for example, deuterium (i.e.,...) with heavier isotopes... 2 H) Greater metabolic stability can provide certain therapeutic advantages (e.g., increased in vivo half-life or reduced dose requirement). In some embodiments, one or more hydrogen atoms are... 2 H or 3 H substitution or one or more carbon atoms being... 13 C- or 14 C-enriched carbon substitution. Positron-emitting isotopes, such as... 15 O、 13 N、 11 C and 18 F can be used in positron emission tomography (PET) studies to examine substrate acceptor occupancy. The preparation of isotopically labeled compounds is known to those skilled in the art. For example, isotopically labeled compounds can typically be prepared by following procedures similar to those disclosed in the compounds of this invention described herein, by replacing non-isotopically labeled reagents with isotopically labeled reagents.

[0335] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Methods and materials used in this disclosure are described herein; other suitable methods and materials known in the art may also be used. The materials, methods, and embodiments described are exemplary and not limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated herein by reference in their entirety. In the event of conflict, this specification, including the definitions, shall prevail.

[0336] definition

[0337] In this application, unless the context clearly indicates otherwise, (i) the term “a” is to be understood as “at least one”; (ii) the term “or” is to be understood as “and / or”; and (iii) the terms “including” and “comprising” are to be understood as covering the listed components or steps, whether presented alone or together with one or more other components or steps.

[0338] As used herein, the terms “about” and “approximately” refer to values ​​that are within 10% above or below the stated value. For example, the term “about 5 nm” indicates a range of 4.5 to 5.5 nm.

[0339] As used herein, the term "administration" refers to the administration of a composition (e.g., a compound or a formulation comprising a compound as described herein) to a subject or system. Administration may be administered to animal subjects (e.g., to humans) via any suitable route. For example, in some embodiments, administration may be via bronchial (including bronchial infusion), oral cavity, enteric, intradermal, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intratumoral, intravenous, intravenous, intracardiac, mucosal, nasal, oral, rectal, subcutaneous, sublingual, local, tracheal (including tracheal infusion), transdermal, vaginal, and vitreous.

[0340] As used in this article, the term "BAF complex" refers to the BRG1 or HRBM-related factor complex in human cells.

[0341] As used herein, the term “BAF complex-related condition” refers to a condition caused by or affected by the activity level of the BAF complex.

[0342] As used herein, the term "BRG1 loss-of-function mutation" refers to a mutation in BRG1 that results in a reduction in protein activity (e.g., a reduction of at least 1% in BRG1 activity, such as a reduction of 2%, 5%, 10%, 25%, 50%, or 100% in BRG1 activity). Exemplary BRG1 loss-of-function mutations include, but are not limited to, homozygous BRG1 mutations and deletions at the C-terminus of BRG1.

[0343] As used herein, the term “BRG1 deficiency syndrome” refers to a syndrome characterized by reduced BRG1 activity (e.g., a reduction of at least 1%, or a reduction of 2%, 5%, 10%, 25%, 50%, or 100% of BRG1 activity).

[0344] The term "cancer" refers to a condition caused by the proliferation of malignant tumor cells, such as tumors, growths, carcinomas, sarcomas, leukemia, and lymphomas.

[0345] As used herein, “combination therapy” or “combination administration” means administering two (or more) different agents or treatments to a subject as part of a defined treatment regimen for a specific disease or condition. The treatment regimen defines the dosage and administration period of each agent such that the effects of the individual agents overlap on the subject. In some embodiments, the delivery of two or more agents is simultaneous or concurrent, and the agents may be co-formulated. In some embodiments, the two or more agents are not co-formulated and are administered sequentially as part of a prescription regimen. In some embodiments, the combination administration of two or more agents or treatments results in a reduction of symptoms or other parameters related to the condition greater than the reduction would be observed with the individual delivery or in the absence of another agent or treatment. The effects of the two treatments may be partially additive, fully additive, or greater than additive (e.g., synergistic). The sequential or substantially simultaneous administration of each therapeutic agent can be achieved via any suitable route, including but not limited to oral, intravenous, intramuscular, and direct absorption through mucosal tissues. The therapeutic agents may be administered via the same or different routes. For example, the first therapeutic agent in a combination may be administered intravenously, while the second therapeutic agent in a combination may be administered orally.

[0346] The term "determination of protein or RNA levels" refers to the direct or indirect detection of protein or RNA by methods known in the art. "Direct determination" means performing a process (e.g., measuring or testing a sample, or, as defined herein, "analyzing a sample") to obtain a physical entity or value. "Indirect determination" means receiving a physical entity or value from another party or source (e.g., a third-party laboratory that directly obtains the physical entity or value). Methods for measuring protein levels generally include, but are not limited to, Western blotting, immunoblotting, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunoprecipitation, immunofluorescence, surface plasmon resonance, chemiluminescence, fluorescence polarization, phosphorescence, immunohistochemistry, matrix-assisted laser desorption / ionization time-of-flight (MALDI-TOF) mass spectrometry, liquid chromatography (LC)-mass spectrometry, microcytology, microscopy, fluorescence-activated cell sorting (FACS), and flow cytometry, as well as determinations based on protein properties, including but not limited to enzyme activity or interactions with other protein couplers. Methods for measuring RNA levels are known in the art, including but not limited to quantitative polymerase chain reaction (qPCR) and Northern blotting analysis.

[0347] The term "reduced level" or "increased level" for protein or RNA refers to a decrease or increase in protein or RNA level compared to a reference substance (e.g., a decrease or increase of approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 150%, 200%, 300%, 400%, 500%, or more; a decrease or increase greater than approximately 10%, approximately...). 15%, approximately 20%, approximately 50%, approximately 75%, approximately 100%, or approximately 200%; a decrease or increase of less than approximately 0.01-fold, approximately 0.02-fold, approximately 0.1-fold, approximately 0.3-fold, approximately 0.5-fold, or approximately 0.8-fold or less; or an increase of more than approximately 1.2-fold, approximately 1.4-fold, approximately 1.5-fold, approximately 1.8-fold, approximately 2.0-fold, approximately 3.0-fold, approximately 3.5-fold, approximately 4.5-fold, approximately 5.0-fold, approximately 10-fold, approximately 15-fold, approximately 20-fold, approximately 30-fold, approximately 40-fold, approximately 50-fold, approximately 100-fold, or approximately 1000-fold or more. Protein levels are expressed in mass / vol (e.g., g / dL, mg / mL, μg / mL, ng / mL) or as a percentage or mass relative to the total protein in the sample.

[0348] The term "reducing the activity of the BAF complex" refers to reducing the activity level associated with the BAF complex or related downstream effects. A non-limiting example of reducing the activity of the BAF complex is Sox2 activation. The activity level of the BAF complex can be measured using any method known in the art, such as that described in Kadoch et al., Cell, 2013, 153, 71-85, which is incorporated herein by reference.

[0349] As used herein, the term "BRM inhibition" refers to blocking or reducing the level or activity of the ATPase-catalyzing binding domain or bromine domain of a protein. BRM inhibition can be determined using methods known in the art, such as BRM ATPase assays, NanoDSF assays, or BRM luciferase cellular assays.

[0350] As used in this article, the term "LXS196," also known as IDE196, refers to a PKC inhibitor with the following structure:

[0351] Or its pharmaceutically acceptable salt.

[0352] As used herein, the term "pharmaceutical composition" means a composition comprising the compounds described herein formulated together with pharmaceutically acceptable excipients and suitable for administration to mammals, such as humans. Typically, pharmaceutical compositions are manufactured or marketed as part of a treatment regimen for treating diseases in mammals, subject to approval by a government regulatory authority. Pharmaceutical compositions may be formulated for oral administration, for example, in unit dosage forms (e.g., tablets, capsules, capsule-shaped tablets, gel caps, or syrups); for topical administration (e.g., as creams, gels, lotions, or ointments); for intravenous administration (e.g., as a sterile solution without emboli and in a solvent system suitable for intravenous use); or in any other pharmaceutically acceptable formulation.

[0353] As used herein, "pharmaceuticalally acceptable excipient" means any component other than the compounds described herein (e.g., a medium capable of suspending or dissolving the active compound) that has substantially non-toxic and non-inflammatory properties in patients. Excipients may include, for example: anti-adhesion agents, antioxidants, adhesives, coatings, compression aids, disintegrants, dyes (colorants), emollients, emulsifiers, fillers (diluents), film-forming agents or coatings, flavoring agents, fragrances, flow enhancers, lubricants, preservatives, printing inks, adsorbents, suspending or dispersing agents, sweeteners, and hydrating water.

[0354] As used herein, the term "pharmaceuticalally acceptable salt" refers to any pharmaceutically acceptable salt of a compound such as any compound of Formula I. Pharmaceutically acceptable salts of any compound described herein may include those that, within reasonable medical judgment, are suitable for use in tissue contact with humans and animals without excessive toxicity, irritation, anaphylactic reactions, and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in: Berge et al., J. Pharmaceutical Sciences 66:1-19, 1977 and Pharmaceutical Salts: Properties, Selection, and Use, (Eds. P.S. Stahl and C.W. Germuth), Wiley-VCH, 2008. Salts may be prepared in situ during the final isolation and purification of the compound described herein, or separately by reacting the free base group with a suitable organic acid.

[0355] The compounds of the present invention may have ionizable groups to enable their preparation as pharmaceutically acceptable salts. These salts may be acid addition salts involving inorganic or organic acids, or, in the case of the acidic form of the compounds of the present invention, the salts may be prepared from inorganic or organic bases. Typically, the compounds are prepared or used as pharmaceutically acceptable salts, which are prepared as addition products of pharmaceutically acceptable acids or bases. Suitable pharmaceutically acceptable acids and bases, and methods for preparing suitable salts, are well known in the art. Salts may be prepared from pharmaceutically acceptable, non-toxic acids and bases, including inorganic and organic acids and bases.

[0356] The term "reference" refers to any useful reference used for comparing protein or RNA levels. A reference can be any sample, standard, standard curve, or level used for comparative purposes. A reference can be a normal reference sample or a reference standard or level. A "reference sample" can be, for example, a control, such as a predetermined negative control value, such as a "normal control," or a previous sample taken from the same subject; a sample from a normal healthy subject, such as normal cells or normal tissue; a sample (e.g., cells or tissue) from a subject without disease; a sample from a subject diagnosed with a disease but not yet treated with the compounds of the present invention; a sample from a subject already treated with the compounds of the present invention; or a sample of purified protein or RNA (e.g., any of those described herein) at a known normal concentration. A "reference standard or level" refers to a value or number derived from a reference sample. A "normal control value" is a predetermined value indicating a non-disease state, such as a value expected in healthy control subjects. Typically, normal control values ​​are expressed as a range ("between X and Y"), a high threshold ("not higher than X"), or a low threshold ("not lower than X"). Subjects with measurements within the normal control range for a particular biomarker are typically referred to as having the biomarker “within the normal limit.” The normal reference standard or level can be a value or number derived from healthy subjects who do not have a disease or condition (e.g., cancer); or subjects who have been treated with the compounds of the present invention. In a preferred embodiment, the reference sample, standard, or level is matched to the sample subject sample by at least one of the following criteria: age, weight, sex, disease stage, and overall health. A standard curve of the levels of purified protein or RNA (e.g., any protein or RNA described herein) within the normal reference range can also be used as a reference.

[0357] As used herein, the term "subject" means any organism to which the compositions according to the invention may be administered, for example, for experimental, diagnostic, preventive, and / or therapeutic purposes. Typical subjects include any animal (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans). A subject may be a person or animal seeking or needing treatment, requesting treatment, receiving treatment, about to receive treatment, or under the care of a professional trained to treat a particular disease or condition.

[0358] As used herein, the terms “treat,” “treated,” or “treating” refer to a therapeutic treatment or any measure aimed at alleviating (reducing) an undesirable physical condition, symptom, or disease, or achieving a beneficial or desired clinical outcome. Beneficial or desired clinical outcomes include, but are not limited to, relief of symptoms; reduction in the severity of a symptom, symptom, or disease; a stable (i.e., non-worsening) state of a condition, symptom, or disease; a delay in the onset or slowing of the progression of a symptom, disorder, or disease; improvement or relief of a symptom, disorder, or disease state (whether partial or complete); improvement in at least one measurable bodily parameter, which may not necessarily be identifiable to the patient; or enhancement or improvement of a symptom, disorder, or disease. Treatment includes causing a clinically significant response without excessive levels of side effects. Treatment also includes prolonged survival compared to expected survival without treatment. The compounds of this invention can also be used for “preventive treatment” or “prevention” of a condition, for example, in subjects at increased risk of developing a condition.

[0359] As used herein, the terms “variant” and “derivative” are used interchangeably and refer to naturally occurring, synthetic, and semi-synthetic analogs of the compounds, peptides, proteins, or other substances described herein. Variants or derivatives of the compounds, peptides, proteins, or other substances described herein may retain or improve the biological activity of the original material.

[0360] Details of one or more embodiments of the invention are set forth in the following description. Other features, objects, and advantages of the invention will be apparent from the specification and claims. Brief description of the attached diagram

[0362] Figure 1 This is a schematic diagram illustrating the inhibition of cell proliferation in several cancer cell lines by an example BRG1 / BRM inhibitor (compound A).

[0363] Figure 2A This is a schematic diagram illustrating the inhibition of cell proliferation in ocular lentigines melanoma cell line 92-1 by BRG1 / BRM inhibitor (compound A), MEK inhibitor (selmetinib), and PKC inhibitor (LXS196).

[0364] Figure 2B This is a schematic diagram illustrating the inhibition of cell proliferation of the ocular lentigines melanoma cell line MP41 by BRG1 / BRM inhibitor (compound A), MEK inhibitor (selmetinib), and PKC inhibitor (LXS196).

[0365] Figure 3 This is a schematic diagram illustrating how a BRG1 / BRM inhibitor (compound B) inhibits cell proliferation in several cancer cell lines.

[0366] Figure 4 This is a schematic diagram illustrating the area under the curve (AUC) calculated from the dose-response curves of cancer cell lines treated with BRG1 / BRM inhibitors.

[0367] Figure 5 This is a schematic diagram illustrating the inhibition of cell proliferation in ocular melanoma and non-small cell lung cancer cell lines by an example BRG1 / BRM inhibitor (compound B).

[0368] Figure 6A This is a schematic diagram illustrating the inhibition of cell proliferation in ocular lentigines melanoma cell line 92-1 by BRG1 / BRM inhibitor (compound B), MEK inhibitor (selmetinib), and PKC inhibitor (LXS196).

[0369] Figure 6B This is a schematic diagram illustrating the inhibition of cell proliferation of the ocular lentigines melanoma cell line MP41 by BRG1 / BRM inhibitor (compound B), MEK inhibitor (selmetinib), and PKC inhibitor (LXS196).

[0370] Figure 7A This is a schematic diagram illustrating the inhibition of cell proliferation in parental and PKC inhibitor-refractory ocular lentigines melanoma cell lines by an example PKC inhibitor (LXS196).

[0371] Figure 7B This is a schematic diagram illustrating the inhibition of cell proliferation in parental and PKC inhibitor-refractory ocular retinoblastoma cell lines by an example BRG1 / BRM inhibitor (compound B).

[0372] Figure 8A This is a schematic diagram illustrating the inhibition of tumor growth in mice transplanted with ocular uvea melanoma cell lines by an example BRG1 / BRM inhibitor (compound C).

[0373] Figure 8B This is a schematic diagram illustrating the tumor size in mice that received an example of an ocular uvea melanoma cell line transplanted and were given a BRG1 / BRM inhibitor (compound C).

[0374] Figure 8C This is a schematic diagram illustrating the weight changes in mice that have been transplanted with an ocular uvea melanoma cell line and given a BRG1 / BRM inhibitor (compound C).

[0375] Detailed description

[0376] This disclosure is characterized by compounds for inhibiting BRG1 and optionally BRG1. These compounds can be used to modulate the activity of the BAF complex, for example, for treating BAF-related conditions, such as cancer (e.g., BRG1-deficiency disorders). Exemplary compounds described herein include compounds having the structure of Formula I or pharmaceutically acceptable salts thereof:

[0377]

[0378] in

[0379] m can be 0, 1, 2, or 3;

[0380] n is 0, 1, 2, 3 or 4;

[0381] X 1 It can be -S-, -SO-, -SO2- or -S(O)(NH)-;

[0382] X 2 For N or CR 8 ;

[0383] R 1 It is hydrogen or an optional substituted C1-C6 alkyl group;

[0384] Each R 2 and each R 3 Independently hydrogen, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl;

[0385] L 1 It is a substituted 9- to 10-membered bicyclic heterocyclic group or a substituted 9- to 10-membered bicyclic heteroaryl group;

[0386] L 2 C3-C does not exist and is an optional substitution. 10 cycloalkyl, optionally substituted C6-C 10 Aryl, optionally substituted 5- to 14-membered heteroaryl or optionally substituted 4- to 14-membered heterocyclic group;

[0387] R 4 It is hydrogen, halogen, optionally substituted C1-C6 alkyl or optionally substituted C3-C 10 cycloalkyl;

[0388] R 5 It is an optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, or optionally substituted amino, and R 6 It is hydrogen, halogen, cyano, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl or optionally substituted C3-C 10 cycloalkyl; or R 5 and R6 Together with the attached atoms, they merge into optionally substituted 5- to 8-membered heterocyclic groups;

[0389] Each R 7 Independently, it is an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 heteroalkyl group, a halogen, or an optionally substituted C3-C... 10 cycloalkyl, optionally substituted C3-C 10 Cycloalkyl C1-C6 alkyl, optionally substituted 5- to 14-membered heteroaryl, optionally substituted 4- to 14-membered heterocyclic, -N(R 7A )2 or -OR 7A , where each R 7A Independently H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 cycloalkyl, optionally substituted C6-C 10 aryl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 4- to 10-membered heterocyclic, or two twin Rs 7A The group, together with the attached atom, merges to form an optionally substituted 5- to 10-membered heteroaryl or an optionally substituted 4- to 10-membered heterocyclic group; or two twin R groups. 7 The group and the attached atom combine to form a carbonyl group;

[0390] R 8 It is hydrogen, halogen, optionally substituted C1-C6 alkyl or optionally substituted C3-C 10 cycloalkyl; and

[0391] R 9 It is hydrogen or halogen;

[0392] Or its pharmaceutically acceptable salt.

[0393] In some embodiments, the compound or a pharmaceutically acceptable salt thereof has the structure of any one of compounds 1-308 in Table 1A. In some embodiments, the compound or a pharmaceutically acceptable salt thereof has the structure of any one of compounds 309-856 in Table 1B.

[0394] Other implementation schemes and exemplary methods for synthesizing these compounds are described herein.

[0395] Drug Use

[0396] The compounds described herein can be used in the methods of this invention, and while not bound by theory, are believed to exert their ability to regulate the level, state, and / or activity of the BAF complex, i.e., by inhibiting the activity of BRG1 and / or BRM proteins within the BAF complex in mammals. BAF complex-related conditions include, but are not limited to, conditions associated with BRG1 loss-of-function mutations.

[0397] One aspect of the invention relates to a method of treating a condition associated with a BRG1 loss-of-function mutation, such as cancer (e.g., non-small cell lung cancer, colorectal cancer, bladder cancer, unknown primary cancer, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, or penile cancer), in a subject in need. In some embodiments, the compound is administered in an amount and at a time that effectively results in one or more (e.g., two or more, three or more, four or more) of the following: (a) reduced tumor size, (b) reduced tumor growth rate, (c) increased tumor cell death, (d) reduced tumor progression, (e) reduced number of metastases, (f) reduced metastasis rate, (g) reduced tumor recurrence, (h) increased subject survival, and (i) increased subject progression-free survival.

[0398] Cancer treatment can result in a reduction in the size or volume of a tumor. For example, after treatment, the tumor size may decrease by 5% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more) relative to its pre-treatment size. Tumor size can be measured by any repeatable means. For example, tumor size can be measured as the diameter of the tumor.

[0399] Cancer treatment can further reduce the number of tumors. For example, after treatment, the number of tumors may be reduced by 5% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more) relative to the number before treatment. The number of tumors can be measured by any repeatable means of measurement; for example, the number of tumors can be measured by counting the tumors that are visible to the naked eye or at a specified magnification (e.g., 2x, 3x, 4x, 5x, 10x, or 50x).

[0400] Cancer treatment can lead to a reduction in the number of metastatic nodules in other tissues or organs far from the site of the primary tumor. For example, after treatment, the number of metastatic nodules may be reduced by 5% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more) relative to the number before treatment. The number of metastatic nodules can be measured by any reproducible method. For example, the number of metastatic nodules can be measured by counting the metastatic nodules visible to the naked eye or at a specified magnification (e.g., 2x, 10x, or 50x).

[0401] Cancer treatment can lead to an increase in the mean survival time of the subject group treated according to the present invention, compared to an untreated subject group. For example, the mean survival time may increase by more than 30 days (more than 60, 90, or 120 days). The increase in mean survival time of the population can be measured in any repeatable manner. The increase in mean survival time of the population can be measured, for example, by calculating the mean survival length of the population after initiating treatment with the compound of the present invention. The increase in mean survival time of the population can also be measured, for example, by calculating the mean survival length of the population after completing the first round of treatment with a pharmaceutically acceptable salt of the present invention.

[0402] Cancer treatment can also lead to a reduction in mortality in the treated population compared to the untreated population. For example, the mortality rate reduction may exceed 2% (e.g., exceed 5%, 10%, or 25%). The reduction in mortality in the treated population can be measured by any reproducible means, for example, by calculating the mean number of disease-related deaths per unit time after the population begins treatment with the pharmaceutically acceptable salt of the present invention. The reduction in population mortality can also be measured, for example, by calculating the mean number of disease-related deaths per unit time after the population completes the first round of treatment with the pharmaceutically acceptable salt of the present invention.

[0403] Exemplary cancers that can be treated by this invention include, but are not limited to, non-small cell lung cancer, small cell lung cancer, colorectal cancer, bladder cancer, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, esophageal and gastric cancer, pancreatic cancer, hepatobiliary cancer, soft tissue sarcoma, ovarian cancer, head and neck cancer, renal cell carcinoma, bone cancer, non-Hodgkin lymphoma, prostate cancer, embryonal tumors, germ cell tumors, cervical cancer, thyroid cancer, salivary gland cancer, gastrointestinal neuroendocrine tumors, uterine sarcoma, gastrointestinal stromal tumors, CNS cancer, thymic tumors, adrenocortical carcinoma, appendix cancer, small bowel cancer, and penile cancer.

[0404] Combination formulations and their uses

[0405] The compounds of the present invention can be combined with one or more therapeutic agents. In particular, the therapeutic agents can be therapeutic agents for treating or preventing any of the cancers described herein.

[0406] combination therapy

[0407] The compounds of this invention can be used alone or in combination with other therapeutic agents (e.g., other agents for treating cancer or related symptoms), or in combination with other types of treatments for cancer. In combination therapy, the dose of one or more therapeutic compounds can be reduced from the standard dose when administered alone. For example, the dose can be determined empirically based on the combination and arrangement of drugs, or derived by isobolographic analysis (e.g., Black et al., Neurology 65:S3-S6, 2005). In this case, when combined, the dose of the compounds should provide a therapeutic effect.

[0408] In some implementations, the second therapeutic agent is a chemotherapeutic agent (e.g., a cytotoxic agent or other chemical compound that can be used to treat cancer). These include alkylating agents, antimetabolites, folic acid analogs, pyrimidine analogs, purine analogs and related inhibitors, vinca alkaloids, epipodophyllotoxin, antibiotics, L-asparaginase, topoisomerase inhibitors, interferon, platinum coordination compounds, anthrone-substituted ureas, methylhydrazine derivatives, adrenocorticotropic hormone inhibitors, adrenocorticotropic hormones, progestins, estrogens, antiestrogens, androgens, antiandrogens, and gonadotropin-releasing hormone analogs. Also included are 5-fluorouracil (5-FU), leucovorin (LV), irinotecan, oxaliplatin, capecitabine, paclitaxel, and docetaxel. Non-limiting examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide; alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethyleneimines and methylamelamines, including altretamine, triethylenemelamine, trietylenephosphoramide, triethiylenethiophosphoramide, and trimethylolomelamine; acetogenins (especially...) These include: bulbatacin and bulbatacinone; camptothecin (including its synthetic analogue topotecan); bryostatin; callystatin; CC-1065 (including its synthetic analogues adozelesin, carzelesin, and bizelesin); cryptophycins (especially cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including its synthetic analogues, KW-2189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictyin; and spongistatin.Nitrogen mustards, such as chlornaphazine, chlorphosphamide, estradiol, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novombhichin, phenesterine, prednimustine, trofosfamide, and uracil mustard; nitrosoureas, such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine; antibiotics, such as enediyne antibiotics (e.g., calicheamicin, especially calicheamicin γ1 and calicheamicin ω1 (see, for example, Agnew, Chem. Intl. Ed.)). Engl.33:183-186(1994)); dynemicin, including dynemicin A; bisphosphonates, such as clophosphonate; esperamicin; and neocarzinostatin chromophores and related chromoproteins (enediyne antibiotic chromophores), aclacinomysins, actinomycin, autramycin, azaserine, bleomycins, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-leucine; (Doxorubicin, including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid Drugs containing citric acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, and zorubicin; antimetabolites, such as methotrexate and 5-fluorouracil (5-FU); and folic acid analogues, such as demofolate, methotrexate, and pteroxate. Eropterin and trimetrexate; purine analogs, such as fludarabine, 6-mercaptopurine, thiamiprine, and thioguanine; pyrimidine analogs, such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, and fluorouridine; and androgens, such as calusterone and dromostanolone. Propionate, epitiostanol, mepitiostane, testolactone; anti-adrenergic drugs, such as aminoglutethimide, mitotane, trilostane; folic acid supplements such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine;Bestrabucil; Bisantrene; Edatraxate; Defofamine; Demecolcine; Diaziquone; Elfomithine; Elliptinium acetate acetate); epomycin; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids, such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinicacid; 2-ethylhydrazine; procarbazine; Polysaccharide complex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin; sizofuran; spirogermanium; tenuazonic acid; triaziquone; 2,2',2”-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A) A) and anguidine; urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactalol; pipebroman; gacytosine; arabinoside (“Ara-C”); cyclophosphamide; thiotepa; taxanes, for example Paclitaxel (Bristol-Myers Squibb Oncology, Princeton, NJ) Cremophor-free, albumin-modified paclitaxel nanoparticle formulations (American Pharmaceutical Partners, Schaumberg, Ill.) and Docetaxel (Rhone-Poulenc Rorer, Antony, France); Chloranbucil; Gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum coordination compounds, such as cisplatin, oxaliplatin and carboplatin; vincristine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; Vinorelbine; novantrone; teniposide; idatraxa; daunomycin; aminopterin; xeloda; ibandronate; irinotecan (e.g., CPT-11); topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids, such as retinoic acid; capecitabine; and pharmaceutically acceptable salts, acids, or derivatives of any of the above. Two or more chemotherapeutic agents may be used in combination with the first therapeutic agent described herein. Suitable dosing regimens for combination chemotherapy are known in the art and described, for example, in Saltz et al. (1999) Proc ASCO 18:233a and Douillard et al. (2000) Lancet 355:1041-7.

[0409] In some embodiments, the second therapeutic agent is a biological agent used for cancer treatment, such as a cytokine (e.g., interferon or interleukin (e.g., IL-2)). In some embodiments, the biological agent is an anti-angiogenic agent, such as an anti-VEGF agent, such as bevacizumab. In some implementations, the biologics are immunoglobulin-based biologics, such as monoclonal antibodies (e.g., humanized antibodies, fully human antibodies, Fc fusion proteins or functional fragments thereof), which activate targets to stimulate anticancer responses or antagonize antigens important to cancer.These drugs include Rituxan; Zenapax; Basiliximab; Synagis; Infliximab; Trastuzumab; Gemtuzumab ozogamicin; Camppath; and Ibritumomab. Tiuxetan; HUMIRA (Adalimumab); Xolair (Omalizumab); Bexxar (Tositumomab-I-131); Raptiva (Efalizumab); Erbitux (Cetuximab); Avastin (Bevacizumab); Tysabri (Natalizumab); Actemra (Tocilizumab); Vectib (Panitumumab); Lucentis (Ranibizumab); Soliris (Eculizumab) Citrozumab; Cimzia (Certolizumabpegol); Simponi (Golimumab); LLARIS (canakinumab); Stelara (Ustekinumab); Arzerra (Ofatumumab); Prolia (Denosumab); Numax (Motavizumab); ABThrax (Raxibacumab); Benlysta (Belimumab); Yervoy (Ipilimumab); Adcetris (Brentuximab) Vedotin; Perjeta (pertuzumab); Kadcyla (ado-trastuzumab emtansine); and Gazyva (obinutuzumab). Antibody-drug conjugates are also included.

[0410] The second agent can be a non-pharmacological treatment. For example, the second agent could be radiation therapy, cryotherapy, hyperthermia, and / or surgical removal of the tumor tissue.

[0411] The second agent can be a checkpoint inhibitor. In one embodiment, the checkpoint inhibitor is an inhibitory antibody (e.g., a monospecific antibody, such as a monoclonal antibody). The antibody can be, for example, humanized or fully human. In some embodiments, the checkpoint inhibitor is a fusion protein, such as an Fc receptor fusion protein. In some embodiments, the checkpoint inhibitor is an agent that interacts with a checkpoint protein, such as an antibody. In some embodiments, the checkpoint inhibitor is an agent that interacts with a ligand of a checkpoint protein, such as an antibody. In some embodiments, the checkpoint inhibitor is a CTLA-4 inhibitor (e.g., an inhibitory antibody or a small molecule inhibitor) (e.g., an anti-CTLA4 antibody, such as ipilimumab / Yervoy or trimemumab). In some embodiments, the checkpoint inhibitor is a PD-1 inhibitor (e.g., an inhibitory antibody or a small molecule inhibitor) (e.g., nivolumab / Pembrolizumab / Pidilizumab / CT-011). In some embodiments, the checkpoint inhibitor is an inhibitor of PDL1 (e.g., an inhibitory antibody or a small molecule inhibitor) (e.g., MPDL3280A / RG7446, MEDI4736, MSB0010718C, BMS 936559). In some embodiments, the checkpoint inhibitor is an inhibitor of PDL2 (e.g., a PDL2 / Ig fusion protein such as AMP) (e.g., an inhibitory antibody or an Fc fusion or small molecule inhibitor). In some embodiments, the checkpoint inhibitor is an inhibitor of B7-H3 (e.g., MGA271), B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, B-7 family ligands, or a combination thereof (e.g., an inhibitory antibody or a small molecule inhibitor).

[0412] In any of the combined embodiments described herein, the first and second therapeutic agents are administered simultaneously or sequentially in either order. The first therapeutic agent may be administered immediately before the second therapeutic agent, or up to 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, or up to 1–7, 1–14, 1–21, or 1–30 days before or after it.

[0413] Pharmaceutical Composition

[0414] The compounds of the present invention are preferably formulated into pharmaceutical compositions for administration to mammals, preferably humans, in a biocompatible form suitable for in vivo administration. Therefore, in one aspect, the present invention provides pharmaceutical compositions comprising the compounds of the present invention mixed with a suitable diluent, carrier, or excipient.

[0415] The compounds of the present invention can be used in the form of free bases, salts, solvates, and prodrugs. All forms are within the scope of the invention. According to the method of the present invention, as those skilled in the art will understand, the compounds, or their salts, solvates, or prodrugs, can be administered to a patient in various forms depending on the chosen route of administration. The compounds of the present invention can be administered, for example, orally, parenterally, orally, sublingually, nasally, rectally, via patch, pump, or transdermally, and pharmaceutical compositions are formulated accordingly. Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary, intrathecal, rectal, and local administration modes. Parenteral administration can be carried out by continuous infusion over a selected time period.

[0416] The compounds of the present invention can be administered orally, for example, with an inert diluent or with an assimilated edible carrier, or encapsulated in hard or soft-shell gelatin capsules, or compressed into tablets, or directly incorporated into food. For oral therapeutic administration, the compounds of the present invention can be mixed with excipients and used in the form of ingestible tablets, lozenges, tablets, capsules, elixirs, suspensions, syrups, and rice paper capsules. The compounds of the present invention can also be administered parenterally. Solutions of the compounds of the present invention can be prepared in water with a suitable surfactant. Under normal storage and use conditions, these formulations may contain preservatives to prevent microbial growth. Conventional methods and ingredients for selecting and preparing suitable formulations are described, for example, in Remington's Pharmaceutical Sciences (2003, 20th edition) and The United States Pharmacopeia: The National Formulary (USP24NF19), published in 1999. Suitable forms of the medicine for injection include sterile aqueous solutions or dispersions and sterile powders for the ad hoc preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and fluid enough to be easily administered via a syringe. Compositions for nasal administration can be conveniently formulated as aerosols, drops, gels, and powders. Aerosol formulations typically comprise a solution or fine suspension of the active ingredient in a physiologically acceptable aqueous or non-aqueous solvent and are usually present in a sterile, single- or multiple-dose form in a sealed container, which may be in cartridge or refill form for use with a nebulizer. Alternatively, the sealed container may be an integral dispensing device, such as a single-dose nasal inhaler or an aerosol dispenser equipped with a metering valve, intended for disposal after use. When the dosage form includes an aerosol dispenser, it will contain a propellant, which may be a compressed gas, such as compressed air, or an organic propellant. Aerosol dosage forms can also be in the form of a pump-nebulizer. Compositions suitable for oral or sublingual administration include tablets, lozenges, and soft lozenges, in which the active ingredient is formulated with a carrier. Compositions for rectal administration are conveniently in suppository form containing a conventional suppository base. The compounds described herein can be administered intratumorally, for example, as intratumoral injection. Intratumoral injection involves direct injection into the tumor's vascular system and is particularly suitable for discrete, solid, accessible tumors. Local, regional, or systemic administration may also be appropriate. The compounds described herein can advantageously be administered to the tumor via a single injection or multiple injections (e.g., at intervals of approximately 1 cm). In the case of surgical intervention, the invention can be used preoperatively, for example, to allow inoperable tumors to be resected. Continuous administration can also be applied where appropriate, for example, by inserting a catheter into the tumor or the tumor's vascular system.

[0417] The compounds of the present invention may be administered to animals, such as humans, alone or in combination with pharmaceutically acceptable carriers as described herein, the proportion of which is determined by the solubility and chemical properties of the compound, the chosen route of administration, and standard pharmaceutical practice.

[0418] dose

[0419] The dosage of the compounds of the present invention and / or compositions comprising the compounds of the present invention can vary depending on many factors, such as the pharmacodynamic properties of the compound; the route of administration; the recipient's age, health, and weight; the nature and severity of symptoms; the frequency of treatment and the type of concurrent treatment (if any); and the clearance rate of the compound in the treated animal. Those skilled in the art can determine an appropriate dosage based on the above factors. The compounds of the present invention can be initially administered at a suitable dosage, which can be adjusted as needed based on clinical response. Generally, satisfactory results can be obtained when the compounds of the present invention are administered to humans at daily doses, for example, from 0.05 mg to 3000 mg. Dosage ranges include, for example, 10-1000 mg.

[0420] Alternatively, the dosage can be calculated using the patient's weight. For example, the dosage of a compound or pharmaceutical composition administered to a patient can be 0.1-100 mg / kg. Example

[0421] The definitions used for the following schemes and other parts of this document are as follows:

[0422] MeCN or ACN acetonitrile

[0423] AIBN (Azobisisobutyronitrile)

[0424] Boc tert-butoxycarbonyl

[0425] t-BuOK potassium tert-butoxide

[0426] DAST diethylaminothiotrifluoride

[0427] DCE dichloroethane

[0428] DCM dichloromethane

[0429] DCPP-2HBF4 1,3-Bis(dicyclohexylphosphino)propane bis(tetrafluoroborate)

[0430] DEA N,N-diethylamine

[0431] DMP Dess-Martin periodinane, 1,1,1-trimethylammonium chloride (DMP)

[0432] (acetoxy)-1,1-dihydro-1,2-DIAD diisopropyl azodicarboxylate

[0433] DIBAL-H Diisopropyl Aluminum Hydrate

[0434] DIEA or DIPEA N,N-diisopropylethylamine

[0435] DMA dimethylacetamide

[0436] DMAP 4-(dimethylamino)pyridine

[0437] DME 1,2-dimethoxyethane

[0438] DMF N,N-dimethylamide

[0439] DMSO (dimethyl sulfoxide)

[0440] dppf bis(diphenylphosphine)ferrocene

[0441] EDCl 1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride

[0442] ESI Electrospray Ionization

[0443] Et3N or TEA triethylamine

[0444] EA (ethyl acetate)

[0445] EtOH (ethanol)

[0446] FA Formic acid

[0447] FCC flash column chromatography

[0448] HATU 2-(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yl)-l,1,3,3-tetramethylisourea

[0449] HCl hydrochloric acid

[0450] HOAc acetic acid

[0451] HOBt Hydroxybenzotriazole

[0452] HPLC (High Performance Liquid Chromatography)

[0453] IPA isopropanol

[0454] LCMS (Liquid Chromatography / Mass Spectrometry)

[0455] m-CPBA 3-chloroperoxybenzoic acid

[0456] MeCN Acetonitrile

[0457] MeI iodomethane

[0458] MeOH (methanol)

[0459] mL

[0460] mmol millimole

[0461] mg

[0462] MHz

[0463] MS mass spectrometry

[0464] MTBE (methyl tert-butyl ether)

[0465] m / z mass / charge ratio

[0466] NBS N-bromosuccinimide

[0467] NIS N-iodosuccinimide

[0468] nm nanometer

[0469] NMR (Nuclear Magnetic Resonance)

[0470] PE petroleum ether

[0471] PhMe Toluene

[0472] ppm (parts per million)

[0473] rt room temperature

[0474] RT Retention Time

[0475] SFC Supercritical Fluid Chromatography

[0476] SPhos Pd G3 (2-Dicyclohexylphosphino-2',6'-dimethoxybiphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II)methanesulfonate

[0477] TBS tert-butyldimethylsilyl

[0478] TBSCl tert-butyldimethylsilyl chloride

[0479] TBDMS tert-butyldimethylsilyl chloride

[0480] TFA (trifluoroacetic acid)

[0481] TFAA (trifluoroacetic anhydride)

[0482] THF Tetrahydrofuran

[0483] TMSCN Trimethylsilyl cyanide

[0484] TosMIC Toluenesulfonylmethyl isocyanate

[0485] Ziram Zinc Dimethyl Dithiocarbamate

[0486] Material

[0487] Unless otherwise stated, all materials were obtained from commercial suppliers and are ready for use without further purification. All reactions involving air- or moisture-sensitive reagents were carried out under a nitrogen atmosphere.

[0488] Table 1C lists the compounds of the present invention prepared using the methods described herein.

[0489] Table 1C. Compounds of the present invention

[0490]

[0491]

[0492]

[0493]

[0494]

[0495]

[0496]

[0497] Material

[0498] Unless otherwise stated, all materials were obtained from commercial suppliers and were ready for use without further purification. All reactions involving air- or moisture-sensitive reagents were carried out under a nitrogen atmosphere.

[0499] Example 1. Preparation of intermediates

[0500] Intermediate 1,1-dioxide of 2,3-dihydro-5H-benzo[e][1,4]oxathihepta-8-carboxylic acid

[0501]

[0502] Step 1: Preparation of methyl 4-bromo-2-mercaptobenzoate

[0503] Sodium sulfide (33.49 g, 429.1 mmol, 18.0 mL) was added to a solution of methyl 4-bromo-2-fluorobenzoate (100 g, 429.12 mmol) in DMF (1 L), and the mixture was stirred at 30 °C for 16 h. The mixture was poured into water (6000 mL), and the pH was adjusted to ~3 with 2N HCl. The mixture was extracted with MTBE (3000 mL x 2). The combined organic phases were washed with brine (3000 mL x 3), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum to give methyl 4-bromo-2-mercaptobenzoate (103 g, crude), a yellow oil, which was used for the next step without further purification.

[0504] 1 H NMR (400MHz, DMSO_d6) δ = 7.91 (d, J = 1.6Hz, 1H), 7.83-7.81 (m, 1H), 7.43-7.40 (m, 1H), 5.58 (br s, 1H), 3.83 (s, 3H) ppm.

[0505] Step 2: Preparation of (4-bromo-2-mercaptophenyl)methanol

[0506] LiAlH4 (15.82 g, 416.82 mmol) was added to a mixture of methyl 4-bromo-2-mercaptobenzoate (103 g, 416.82 mmol) in THF (1000 mL) at 0 °C under a N2 atmosphere. The mixture was stirred at 0 °C for 1 hr. The mixture was poured into 1N HCl (2000 mL) and extracted with EtOAc (2000 mL x 2). The combined organic phases were washed with brine (2000 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum to give (4-bromo-2-mercaptophenyl)methanol (88 g, crude), a yellow oil, which was used for the next step without further purification. 1 H NMR (400MHz, DMSO_d6) δ = 7.59 (s, 1H), 7.32 (d, J = 1.2Hz, 2H), 5.56-5.36 (m, 2H), 4.39 (s, 2H) ppm.

[0507] Step 3: Preparation of (4-bromo-2-(vinylthio)phenyl)methanol and (4-bromo-2-((2-bromoethyl)thio)phenyl)methanol

[0508] To a mixture of (4-bromo-2-mercaptophenyl)methanol (85 g, 387.95 mmol) in DMF (1700 mL), K₂CO₃ (160.9 g, 1.16 mol) and 1,2-dibromoethane (218.6 g, 1.16 mol, 87.8 mL) were added, and the mixture was stirred at 25 °C for 1 hour. The mixture was then stirred at 70 °C for another 24 hours. The reaction mixture was poured into saturated NH₄Cl (10 L) and extracted with EA (3000 mL x 2). The combined organic layers were washed with brine (4000 mL x 2), dried over Na₂SO₄, filtered, and the filtrate was evaporated to dryness. The residue was purified by silica gel column chromatography (PE / EA = 50 / 1–5 / 1). Vacuum concentration fractionation yielded (4-bromo-2-(vinylthio)phenyl)methanol (33.5 g, 136.66 mmol, 35% yield) and (4-bromo-2-((2-bromoethyl)thio)phenyl)methanol (10 g, 30.67 mmol, 8% yield), both as yellow oils.

[0509] (4-Bromo-2-(vinylthio)phenyl)methanol: 1 H NMR (400MHz, CDCl3) δ = 7.55 (s, 1H), 7.45 (d, J = 2Hz, 1H), 7.43 (d, J = 2Hz, 1H), 6. 49-6.42(m,1H),5.45(d,J=9.6Hz,1H),5.32(d,J=10.4Hz,1H),4.73(s,2H)ppm;

[0510] (4-Bromo-2-((2-Bromoethyl)thio)phenyl)methanol: 1 H NMR (400MHz, CDCl3) δ = 7.46 (s, 1H), 7.33 (d, J = 2Hz, 1H), 7.09 (d, J = 2Hz, 1H), 6.67 (s, 2H), 3.41-3.38 (m, 2H), 3.25-3.23 (m, 1H) ppm.

[0511] Step 4: Preparation of (4-bromo-2-(vinylsulfonyl)phenyl)methanol

[0512] Add to a mixture of (4-bromo-2-(vinylthio)phenyl)methanol (35.5 g, 144.82 mmol) in MeOH (350 mL) and H₂O (350 mL) (133.54 g, 217.23 mmol) The mixture was stirred at 25 °C for 2 h. Water (1500 mL) was added, and the mixture was extracted with EtOAc (1500 mL x 2). The combined organic phases were washed with brine (1000 mL x 2), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum to give (4-bromo-2-vinylsulfonyl-phenyl)methanol (38.5 g, crude), a yellow solid, which was used for the next step without further purification.

[0513] 1 H NMR(400MHz, DMSO_d6)δ=7.98-7.95(m,2H),7.77-7.75(m,1H),7.22-7.15(m,1H),6. 43-6.39(m,1H),6.31(d,J=10.0Hz,1H),5.62-5.59(m,1H),4.75(d,J=5.2Hz,2H)ppm.

[0514] Step 5: Preparation of 8-bromo-2,3-dihydro-5H-benzo[e][1,4]oxathiheptazone 1,1-dioxide

[0515] NaH (11.11 g, 277.84 mmol, 60% purity) was added to a mixture of (4-bromo-2-vinylsulfonyl-phenyl)methanol (38.5 g, 138.9 mmol) in DMF (1000 mL) at 0 °C under a nitrogen atmosphere. The mixture was stirred at 0 °C for 1 hr. The reaction mixture was poured into saturated NH4Cl (2 L) and extracted with EA (2000 mL * 2). The combined organic phases were washed with (2000 mL) brine, dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue (SiO2, PE:EtOAc = 50:1-5:1) was purified by column chromatography and concentrated under vacuum to give 8-bromo-2,3-dihydro-5H-benzo[e][1,4]oxathihexane 1,1-dioxide (26.5 g, 95.62 mmol, 69% yield) as a white solid. 1 H NMR (400MHz, DMSO_d6) δ = 7.99 (d, J = 2.0Hz, 1H), 7.92-7.90 (m, 1H), 7.55 (d, J = 8.0Hz, 1H), 4.88 (s, 2H), 4.20-4.17 (m, 2H), 3.68-3.66 (m, 2H) ppm.

[0516] Step 6: Preparation of 1,1-dioxide of 2,3-dihydro-5H-benzo[e][1,4]oxathihepta-8-carboxylic acid (intermediate 1)

[0517] To a mixture of 8-bromo-2,3-dihydro-5H-benzo[e][1,4]oxathiheptazone 1,1-dioxide (8.8 g, 31.75 mmol) in DMSO (90 mL) and H₂O (9 mL), 1,3-bis(dicyclohexylphosphino)propane bis(tetrafluoroborate) (3.89 g, 6.35 mmol), K₂CO₃ (6.58 g, 47.63 mmol), and Pd(OAc)₂ (712.90 mg, 3.18 mmol) were added. The mixture was purged three times with CO and then stirred at 100 °C under a CO atmosphere (15 psi) for 4 h. Water (3000 mL) was added, and the mixture was extracted with EtOAc (500 mL x 2), then the organic phase was discarded. The pH of the aqueous layer was adjusted to ~3 with 1 N HCl. The mixture was then extracted with EA (500 mL x 5). The combined organic phases were washed with brine (2000 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under vacuum. The crude product was washed with MTBE (20 mL x 2), filtered, and the filter cake was evaporated to dryness to give 1,1-dioxide of 2,3-dihydro-5H-benzo[e][1,4]oxathiheptacosan-8-carboxylic acid (15 g, 61.92 mmol, 65% yield) as a white solid.

[0518] 1 H NMR (400MHz, DMSO-d6) δ = 8.43 (s, 1H), 8.20-8.18 (m, 1H), 7.72-7.70 (m, 1H), 4.96 (s, 2H), 4.23-4.20 (m, 2H), 3.67-3.66 (m, 2H) ppm.

[0519] Intermediate 2.3,5-dihydro-2H-[1,4]oxathiheptan[6,5-b]pyridine-8-carboxylic acid 1,1-dioxide

[0520]

[0521] Step 1: Preparation of methyl 5-bromo-3-mercaptopyridinecarboxylate

[0522] Na₂S (333.49 mg, 4.27 mmol) was added to a solution of methyl 5-bromo-3-fluoro-pyridine-2-carboxylate (1 g, 4.27 mmol) in DMF (10 mL). The mixture was stirred at 25 °C for 2 h. Three identical batches were combined and purified together. The mixture was diluted with water (50 mL) and adjusted to pH 5 with 1 N HCl aqueous solution. The mixture was extracted with EA (50 mL x 2). The combined organic layers were washed with brine (50 mL x 2), dried over anhydrous Na₂SO₄, and concentrated to give methyl 5-bromo-3-mercaptopyridinecarboxylate (3.3 g, crude), as a brown oil. LCMS (ESI) m / z: [M+H] + =247.8 / 249.8.

[0523] Step 2: Preparation of (5-bromo-3-mercaptopyridin-2-yl)methanol

[0524] LiAlH4 (504.8 mg, 13.30 mmol) was added to a solution of methyl 5-bromo-3-mercaptopyridinecarboxylate (3.3 g, 13.30 mmol) in THF (33 mL) at 0 °C. The mixture was stirred at 25 °C for 2 h. The mixture was diluted with water (100 mL) and adjusted to pH 6 with 1 N HCl aqueous solution. The mixture was then extracted with EA (100 mL x 2). The combined organic layers were dried over anhydrous Na2SO4 and concentrated to give (5-bromo-3-mercaptopyridine-2-yl)methanol (1.66 g, 7.54 mmol) as a brown oil.

[0525] LCMS(ESI)m / z:[M+H] + =219.8 / 221.8.

[0526] Step 3: Preparation of (5-bromo-3-(vinylthio)pyridin-2-yl)methanol

[0527] To a solution of (5-bromo-3-mercaptopyridin-2-yl)methanol (1.66 g, 7.54 mmol) in DMF (15 mL), K₂CO₃ (3.13 g, 22.63 mmol) and 1,2-dibromoethane (7.08 g, 37.71 mmol, 2.85 mL) were added. The mixture was stirred at 60 °C for 12 h. The mixture was diluted with water (100 mL) and extracted with EA (100 mL x 2). The combined starch organic layers were dried over anhydrous Na₂SO₄ and concentrated to give the residue. The residue was purified by silica gel rapid chromatography (…). 20g Silica gel rapid column chromatography, 0–100% ethyl acetate / petroleum ether eluent. Concentrate the eluent to give (5-bromo-3-(vinylthio)pyridin-2-yl)methanol (600 mg, 2.44 mmol, 32% yield) as a brown oil.

[0528] LCMS(ESI)m / z:[M+H] + =245.9 / 247.9;

[0529] 1 HNMR (400MHz, DMSO-d6) δ = 8.53 (d, J = 2.0Hz, 1H), 7.92 (d, J = 2.0Hz, 1H), 6.81- 6.74(m,1H),5.64-5.51(m,2H),5.32-5.29(m,1H),4.54(d,J=6.0Hz,2H)ppm.

[0530] Step 4: Preparation of (5-bromo-3-(vinylsulfinyl)pyridin-2-yl)methanol

[0531] Slowly add (600 mg, 2.44 mmol) of 5-bromo-3-(vinylthio)pyridin-2-yl)methanol (in 6 mL of MeOH) to a solution of 5-bromo-3-(vinylthio)pyridin-2-yl)methanol in 6 mL of water at 0 °C. (824.27 mg, 1.34 mmol). The mixture was stirred at 25 °C for 1 hr. The mixture was quenched with a saturated aqueous solution of Na₂SO₃ (30 mL) and extracted with EA (30 mL x 2). The combined organic layers were dried over anhydrous Na₂SO₄ and concentrated to give the residue. The residue was purified by silica gel rapid chromatography. 12g Silica gel rapid column chromatography, 0–100% ethyl acetate / petroleum ether eluent. Concentrate the eluent to give (5-bromo-3-(vinylsulfinyl)pyridin-2-yl)methanol (500 mg, 1.91 mmol, 78.25% yield) as a colorless oil.

[0532] 1 HNMR (400MHz, DMSO-d6) δ = 8.74 (d, J = 2.4Hz, 1H), 8.17 (d, J = 2.0Hz, 1H), 7.18-7.12 (m, 1 H),6.09-6.02(m,2H),5.95(d,J=9.6Hz,1H),4.85-4.78(m,1H),4.73-4.66(m,1H)ppm.

[0533] Step 5: Preparation of 1-oxide of 8-bromo-3,5-dihydro-2H-[1,4]oxathiheptan[6,5-b]pyridine

[0534] NaH (152.59 mg, 3.81 mmol, 60% purity) was added to a solution of (5-bromo-3-(vinylsulfinyl)pyridin-2-yl)methanol (500 mg, 1.91 mmol) in DMF (5 mL) at 0 °C. The mixture was stirred at 0 °C for 2 h. The mixture was quenched with saturated NH4Cl aqueous solution (30 mL) and extracted with EA (30 mL x 2). The combined organic layers were dried over Na2SO4-free water, concentrated, and the residue was obtained. The residue was purified by silica gel rapid chromatography. 12g Silica gel rapid column chromatography, 0–10% ethyl acetate / petroleum ether eluent. Concentrate the eluent to give 8-bromo-3,5-dihydro-2H-[1,4]oxathiaheptano[6,5-b]pyridine 1-oxide (350 mg, 1.34 mmol, 70% yield), as a colorless oil.

[0535] 1 HNMR (400MHz, DMSO-d6) δ = 8.75 (d, J = 2.4Hz, 1H), 8.19 (d, J = 2.0Hz, 1H), 4.91-4.74 (m ,2H),4.43-4.34(m,1H),4.21-4.18(m,1H),3.65-3.56(m,1H),3.49-3.44(m,1H)ppm.

[0536] Step 6: Preparation of 1-oxide of 3,5-dihydro-2H-[1,4]oxathiheptan[6,5-b]pyridine-8-carboxylic acid

[0537] To a solution of 8-bromo-3,5-dihydro-2H-[1,4]oxathiheptaphyllo[6,5-b]pyridine 1-oxide (350 mg, 1.34 mmol) in DMSO (4 mL) and water (120.27 mg, 6.68 mmol, 120.27 μL), 1,3-bis(dicyclohexylphosphino)propane bis(tetrafluoroborate) (81.75 mg, 133.52 μmol), K₂CO₃ (276.82 mg, 2.00 mmol), and Pd(OAc)₂ (29.98 mg, 133.52 μmol) were added. The mixture was degassed and purged three times with CO. The mixture was stirred at 100 °C under a CO atmosphere (15 psi) for 12 h. The mixture was filtered and washed with DMSO (2 mL) and water (2 mL). The filtrate was then adjusted to pH 6 with 1 N HCl aqueous solution. The filtrate was purified by reversed-phase HPLC (0.1% FA conditions). The eluent was concentrated to remove ACN, and lyophilized to give 3,5-dihydro-2H-[1,4]oxathiaheptano[6,5-b]pyridine-8-carboxylic acid 1-oxide (70 mg, 0.262 mmol, 20% yield) as a white solid.

[0538] LCMS(ESI)m / z:[M+H] + =227.9;

[0539] 1 HNMR (400MHz, DMSO-d6) δ = 9.03 (d, J = 2.0Hz, 1H), 8.51 (d, J = 2.0Hz, 1H), 5.01-4.88 (m ,2H),4.43-4.41(m,1H),4.18-4.15(m,1H),3.63-3.62(m,2H),3.52-3.48(m,2H)ppm.

[0540] Step 7: Preparation of 1,1-dioxide (intermediate 2) of 3,5-dihydro-2H-[1,4]oxathiheptan[6,5-b]pyridine-8-carboxylic acid

[0541] Add 70 mg (0.309 mmol) of 1-oxide of 3,5-dihydro-2H-[1,4]oxathiaheptano[6,5-b]pyridine-8-carboxylic acid (1-oxide) in 0.7 mL of MeOH to a solution of 1-oxide in 0.7 mL of water at 0 °C. (284.07 mg, 462.07 μmol). The mixture was stirred at 25 °C for 1 hr. The mixture was then filtered. The filter cake was washed with MeOH (5 mL). The filtrate was then quenched with saturated Na₂SO₃ solution. The solution was then purified by reversed-phase HPLC (0.1% FA conditions). The eluent was concentrated to remove ACN, and lyophilized to give 1,1-dioxide of 3,5-dihydro-2H-[1,4]oxathiaheptano[6,5-b]pyridine-8-carboxylic acid (36 mg, 136.16 μmol, 44% yield) as a white solid. LCMS (ESI) m / z: [M+H] + =243.9.

[0542] Intermediate 3. 3,4-Dihydro-2H-benzo[b][1,4]oxathiahepta-7-carboxylic acid 5,5-dioxide

[0543]

[0544] Step 1: Preparation of 3-chlorosulfonyl-4-hydroxybenzoic acid

[0545] 4-Hydroxybenzoic acid (5.5 g, 39.82 mmol) was added stepwise to an HSO3Cl solution (31 mL). The mixture was stirred at 20 °C for 16 h. Ice water (300 mL) was slowly added dropwise to the reaction mixture. The mixture was extracted with ethyl acetate (100 mL x 3). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, and concentrated under vacuum to give the residue. The crude product was ground together with PE (30 mL) at 20 °C for 30 min to give 3-chlorosulfonyl-4-hydroxybenzoic acid (4.5 g, 13.72 mmol, 74% yield) as a white solid. 1 HNMR (400MHz, DMSO-d6) δ = 8.09-8.06 (m, 1H), 7.82-7.75 (m, 1H), 6.89-6.81 (m, 1H) ppm.

[0546] Step 2: Preparation of 4-hydroxy-3-mercaptobenzoic acid

[0547] PPh3 (3.88 g, 14.79 mmol) was added fractionally to a solution of 3-chlorosulfonyl-4-hydroxybenzoic acid (1 g, 4.23 mmol) in toluene (20 mL). The mixture was stirred at 90 °C for 2 h. The reaction was stopped by adding 10% NaOH solution (20 mL). The mixture was extracted with ethyl acetate (20 mL x 3). The aqueous phase was adjusted to pH 2 with 1 N HCl. The mixture was extracted with ethyl acetate (20 mL x 3). The combined organic layers were dried over Na2SO4 and concentrated under vacuum to give 4-hydroxy-3-mercaptobenzoic acid (0.62 g, 3.64 mmol, 86.21% yield) as a yellow oil.

[0548] 1 H NMR (400MHz, DMSO-d6) δ = 12.34 (s, 1H), 7.89-7.83 (m, 1H), 7.61-7.52 (m, 1H), 6.90-6.83 (m, 1H), 5.01 (s, 1H) ppm.

[0549] Step 3: Preparation of methyl 4-hydroxy-3-mercaptobenzoate

[0550] H₂SO₄ (352.84 mg, 3.53 mmol, 191.76 μL, 98% purity) was added dropwise to a solution of 4-hydroxy-3-mercaptobenzoic acid (0.6 g, 3.53 mmol) in MeOH (5 mL). The mixture was stirred at 70 °C for 40 h. The reaction was stopped by adding water (20 mL). The mixture was extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na₂SO₄, and concentrated under vacuum to give methyl 4-hydroxy-3-mercaptobenzoate (0.6 g, crude) as a white solid.

[0551] 1 H NMR (400MHz, DMSO-d6) δ = 11.30 (s, 1H), 8.07-8.01 (m, 1H), 7.77-7.71 (m, 1H), 6.99-6.95 (m, 1H), 3.80-3.78 (m, 3H) ppm.

[0552] Step 4: Preparation of methyl 3,4-dihydro-2H-benzo[b][1,4]oxathiheptaphylline-7-carboxylate

[0553] Cs₂CO₃ (884.36 mg, 2.71 mmol) was added to a solution of methyl 4-hydroxy-3-mercaptobenzoate (0.1 g, 542.85 μmol, 1 eq) in DMF (5 mL), followed by dropwise addition of 1,3-dibromopropane (109.6 mg, 0.543 mmol, 55 μL). The mixture was stirred at 20 °C for 2 h. The reaction was stopped by adding water (20 mL). The mixture was extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na₂SO₄, and concentrated under vacuum to give the residue. The residue (SiO2, petroleum ether: ethyl acetate = 1:1) was purified by preparative-TLC, and the eluent was concentrated under vacuum to give methyl 3,4-dihydro-2H-benzo[b][1,4]oxathiaheptacoine-7-carboxylate (65 mg, 0.274 mmol, 51% yield), which was a yellow oil.

[0554] LCMS(ESI)m / z:[M+H] + =225.1;

[0555] 1 H NMR(400MHz, CDCl3)δ=8.09-8.02(m,1H),7.83-7.74(m,1H),7.03-6.94(m,1H), 4.45-4.34(m,2H),3.93-3.84(m,3H),3.10-2.98(m,2H),2.34-2.22(m,2H)ppm.

[0556] Step 5: Preparation of methyl 3,4-dihydro-2H-benzo[b][1,4]oxathiheptaenoic acid-7-carboxylate 5,5-dioxide

[0557] Add to a mixture of methyl 3,4-dihydro-2H-1,5-benzoxazothionein-7-carboxylate (60 mg, 267.53 μmol) in MeOH (5 mL) and H₂O (5 mL) (493.40 mg, 802.58 μmol), and the mixture was then stirred at 20 °C for 16 h. The reaction was quenched by adding saturated Na₂SO₃ (30 mL). The mixture was extracted with DCM (30 mL x 5). The combined organic layers were dried with Na₂SO₄ and concentrated under vacuum to give methyl 3,4-dihydro-2H-benzo[b][1,4]oxathiheptaenoyl-7-carboxylate 5,5-dioxide (66 mg, 0.257 mmol, 96% yield), as a yellow oil.

[0558] 1H NMR(400MHz, CDCl3)δ=8.73-8.60(m,1H),8.32-8.16(m,1H),7.26-7.23(m,1H), 4.43-4.30(m,2H),3.97-3.91(m,3H),3.48-3.36(m,2H),2.53-2.41(m,2H)ppm.

[0559] Step 6: Preparation of 5,5-dioxide of 3,4-dihydro-2H-benzo[b][1,4]oxathihepta-7-carboxylic acid (intermediate 3)

[0560] NaOH (30.44 mg, 0.761 mmol) was gradually added to a mixture of methyl 3,4-dihydro-2H-benzo[b][1,4]oxathihexane-7-carboxylic acid 5,5-dioxide (65 mg, 0.254 mmol) in MeOH (3 mL) and H₂O (3 mL), and the mixture was stirred at 20 °C for 2 h. The reaction system was concentrated under vacuum to obtain a residue. The residue was partitioned with EA (10 mL) and 1N NaOH solution (10 mL). The aqueous layer was adjusted to pH 1 with 1N HCl solution and extracted with EA (10 mL x 3). The combined organic phases were concentrated under vacuum to give 3,4-dihydro-2H-benzo[b][1,4]oxathihexane-7-carboxylic acid 5,5-dioxide (60 mg, 0.248 mmol, 98% yield) as a yellow solid. LCMS (ESI) m / z: [M+Na] + =265.2;

[0561] 1 H NMR (400MHz, DMSO-d6) δ=8.42-8.30(m,1H),8.21-8.10(m,1H),7.41-7.29(m,1H),4.34-4.22(m,2H),2.31-2.22(m,2H),1.81-1.71(m,2H)ppm.

[0562] Intermediate 4. 6-Chloro-2,3-dihydro-5H-benzo[e][1,4]oxathihepta-8-carboxylic acid 1,1-dioxide

[0563] Step 1: Preparation of methyl 4-bromo-2-chloro-6-fluorobenzoate

[0564] To a solution of 4-bromo-2-chloro-6-fluorobenzoic acid (10 g, 39.46 mmol) in MeOH (90 mL), concentrated H₂SO₄ (18.4 g, 187.60 mmol, 10 mL) was slowly added, and the mixture was stirred at 70 °C for 8 h. The mixture was concentrated under vacuum to remove some of the MeOH, then poured into saturated NaHCO₃ (200 mL), and extracted with EA (200 mL x 2). The combined organic layers were washed with brine (100 mL x 2), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give methyl 4-bromo-2-chloro-6-fluorobenzoate (9.2 g, crude), a colorless oil, which was used directly for the next step.

[0565] 1 H NMR (400MHz, DMSO-d6) δ = 7.87-7.76 (m, 2H), 3.91 (s, 3H) ppm.

[0566] Step 2: Preparation of methyl 4-bromo-2-chloro-6-mercaptobenzoate

[0567] To a solution of methyl 4-bromo-2-chloro-6-fluorobenzoate (7.2 g, 26.92 mmol) in DMF (72 mL), Na₂S (2.10 g, 26.92 mmol) was added, and the mixture was stirred at 25 °C for 2 h. The mixture was diluted with water (300 mL), and then acidified to pH 3 with 1 N HCl solution. Extraction was performed with EA (200 mL x 2). The combined organic layers were washed with brine (250 mL x 2), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give methyl 4-bromo-2-chloro-6-mercaptobenzoate (7 g, crude), as a yellow oil. The crude product was used directly in the next step.

[0568] Step 3: Preparation of (4-bromo-2-chloro-6-mercaptophenyl)methanol

[0569] LiAlH4 (1.33 g, 35.16 mmol) was added to a mixture of methyl 4-bromo-2-chloro-6-mercaptobenzoate (9 g, 31.97 mmol) in THF (90 mL) at 0 °C, and the mixture was stirred at 0 °C for 1 hr. The mixture was poured into HCl (1 N, 200 mL) and then extracted with EA (250 mL x 2). The combined organic layers were washed with brine (200 mL x 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to give (4-bromo-2-chloro-6-mercaptophenyl)methanol (5.8 g, crude) as a colorless oil.

[0570] Step 4: Preparation of (4-bromo-2-chloro-6-(vinylthio)phenyl)methanol

[0571] To a mixture of (4-bromo-2-chloro-6-mercaptophenyl)methanol (5.7 g, 22.48 mmol) in DMF (110 mL), K₂CO₃ (9.32 g, 67.44 mmol) and 1,2-dibromoethane (21.12 g, 112.41 mmol, 8.5 mL) were added, and the mixture was stirred at 25 °C for 12 h. The mixture was poured into water (200 mL) and extracted with EA (100 mL). The combined organic layers were dried over Na₂SO₄, filtered, and concentrated under vacuum to give the residue. The residue was purified by column chromatography (PE / EA = 10 / 1-1:1, SiO₂), and the eluent was evaporated to give (4-bromo-2-chloro-6-(vinylthio)phenyl)methanol (2.7 g, 9.66 mmol, 43% yield) as a colorless oil.

[0572] 1 H NMR (400MHz, DMSO-d6) δ = 7.65 (d, J = 2.0Hz, 1H), 7.42 (d, J = 2.0Hz, 1H), 6.78-6.71 (m, 1H), 5.61 -5.52(m,2H),5.25-5.23(m,1H),4.62(d,J=5.2Hz,2H)ppm.

[0573] Step 5: Preparation of methyl 3-chloro-4-(hydroxymethyl)-5-(vinylthio)benzoate

[0574] Pd(OAc)₂ (80.30 mg, 357.68 μmol) and XPhos (341 mg, 0.715 mmol) were added to a mixture of (1000 mg, 3.58 mmol) in MeOH (20 mL) and TEA (10 mL). The mixture was then degassed, purged three times with CO (15 psi), and stirred at 70 °C under a CO atmosphere (15 psi) for 8 h. The mixture was diluted with water (20 mL), extracted with EA (15 mL x 3), washed with brine (30 mL), dried over Na₂SO₄, filtered, and concentrated under vacuum to give the crude product. The residue was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 20 / 1-5 / 1). Vacuum concentration fractionation yielded methyl 3-chloro-4-(hydroxymethyl)-5-(vinylthio)benzoate (650 mg, 2.36 mmol, 66% yield), a white solid.

[0575] 1H NMR (400MHz, DMSO-d6) δ = 7.83 (d, J = 1.6Hz, 1H), 7.80 (d, J = 1.6Hz, 1H), 6.74-6.67 (m ,1H),5.62-5.51(m,2H),5.36-5.33(m,1H),4.70(d,J=5.2Hz,2H),3.87(s,3H)ppm.

[0576] Step 6: Preparation of methyl 3-chloro-4-(hydroxymethyl)-5-(vinylsulfonyl)benzoate

[0577] Add to a mixture of methyl 3-chloro-4-(hydroxymethyl)-5-(vinylthio)benzoate (500 mg, 1.93 mmol) in H₂O (5 mL) and MeOH (5 mL) (3.56 g, 5.80 mmol) The mixture was stirred at 25 °C for 1 hr. The mixture was diluted with water (200 mL), then extracted with EA (250 mL x 2). The combined organic solutions were washed with saturated Na₂SO₃ (150 mL x 2) and brine (100 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give methyl 3-chloro-4-(hydroxymethyl)-5-vinylsulfonylbenzoate (560 mg, crude), as a yellow oil. LCMS (ESI) m / z: [M+H] + =273.0; 1 HNMR (400MHz, DMSO-d6) δ = 8.42 (d, J = 1.6 Hz, 1H), 8.27 ( d, J = 1.6 Hz, 1H), 7.34-7.27 ( m,1H),6.47-6.31(m,2H),5.52-5.49(m,2H),4.98(d,J=5.2Hz,2H),3.91(s,3H)ppm.

[0578] Step 7: Preparation of 1,1-dioxide of 6-chloro-2,3-dihydro-5H-benzo[e][1,4]oxathihepta-8-carboxylic acid (intermediate 4)

[0579] NaH (154.09 mg, 3.85 mmol, 60% purity) was added to a mixture of methyl 3-chloro-4-(hydroxymethyl)-5-vinylsulfonylbenzoate (560 mg, 1.93 mmol) in THF (18 mL) at 0 °C, and the mixture was stirred at 0 °C for 1 hr. The mixture was diluted with water (10 mL) and MeOH (5 mL), and then stirred at 25 °C for 15 min. The resulting mixture was diluted with water (100 mL), acidified to pH 2 with HCl (1N), and the solution was extracted with EA (150 mL x 2). The combined organic layers were washed with brine (200 mL), dried with Na2SO4, filtered, and concentrated under reduced pressure to give 1,1-dioxide of 6-chloro-2,3-dihydro-5H-benzo[e][1,4]oxathiheptacosan-8-carboxylic acid (300 mg, crude) as a white solid.

[0580] 1 H NMR (400MHz, DMSO-d6) δ = 13.91-13.84 (m, 1H), 8.38 (d, J = 1.6Hz, 1H), 8.22 (d ,J=1.6Hz,1H),5.19(s,2H),4.23-4.21(m,2H),3.79-3.77-3.74(m,2H)ppm.

[0581] Intermediate 5: 6-Fluoro-2,3-dihydro-5H-benzo[e][1,4]oxathihepta-8-carboxylic acid 1,1-dioxide

[0582]

[0583] Step 1: Preparation of methyl 4-bromo-2-fluoro-6-((4-methoxybenzyl)thio)benzoate

[0584] Cs₂CO₃ (12.98 g, 39.84 mmol) was added to a mixture of methyl 4-bromo-2,6-difluorobenzoate (5 g, 19.92 mmol, 2.77 mL) in DMF (50 mL), and the mixture was stirred at 60 °C for 2 h. The mixture was diluted with water (400 mL), extracted with EA (200 mL x 3), the combined organic layers were washed with brine (200 mL x 2), dried over Na₂SO₄, filtered, and concentrated under vacuum to give methyl 4-bromo-2-fluoro-6-((4-methoxybenzyl)thio)benzoate (9 g, crude), as a yellow oil, which was used directly for the next step.

[0585] 1H NMR (400MHz, DMSO-d6) δ=7.54-7.51(m,2H),7.29-7.26(m,2H),6.90-6.87(m,2H),4.29(s,2H),3.83(s,3H),3.73-3.72(m,3H)ppm.

[0586] Step 2: Preparation of methyl 4-bromo-2-fluoro-6-mercaptobenzoate

[0587] A mixture of methyl 4-bromo-2-fluoro-6-((4-methoxybenzyl)thio)benzoate (9 g, 23.36 mmol) in TFA (138.60 g, 1.22 mol, 90 mL) was stirred at 60 °C for 2 h. The mixture was evaporated and then neutralized to pH 7 with saturated NaHCO3. The mixture was then extracted with EA (200 mL). The organic layer was separated and dried with anhydrous Na2SO4. The organic phase was concentrated under vacuum to give methyl 4-bromo-2-fluoro-6-mercaptobenzoate (6 g, crude), a yellow oil, which was used directly for the next step.

[0588] 1 H NMR (400MHz, DMSO-d6) δ = 7.77-7.61 (m, 2H), 3.93 (s, 3H) ppm.

[0589] Step 3: Preparation of (4-bromo-2-fluoro-6-mercaptophenyl)methanol

[0590] To a mixture of methyl 4-bromo-2-fluoro-6-mercaptobenzoate (3.4 g, 12.83 mmol) in THF (34 mL), LiAlH4 (535.5 mg, 14.11 mmol) was added, and the mixture was stirred at 0 °C for 1 hr. The mixture was quenched with 1 N HCl (100 mL) and extracted with EA (50 mL). The organic layer was separated, dried over anhydrous Na2SO4, filtered, and concentrated under vacuum to give (4-bromo-2-fluoro-6-mercaptophenyl)methanol (3 g, crude), a yellow oil, which was used directly for the next step.

[0591] 1 H NMR (400MHz, DMSO-d6) δ = 7.51 (s, 1H), 7.32-7.24 (m, 1H), 4.45 (d, J = 1.2Hz, 2H).

[0592] Step 4: Preparation of (4-bromo-2-fluoro-6-(vinylthio)phenyl)methanol

[0593] To a mixture of (4-bromo-2-fluoro-6-mercaptophenyl)methanol (3 g, 12.65 mmol), K₂CO₃ (5.25 g, 37.96 mmol), and DMF (60 mL), 1,2-dibromoethane (11.89 g, 63.27 mmol) was added, and the mixture was stirred at 25 °C for 15 h. The mixture was poured into water (200 mL) and extracted with EA (100 mL). The combined organic layers were dried over Na₂SO₄, filtered, and concentrated under vacuum to obtain the residue. The residue was purified by column chromatography (PE / EA = 10 / 1, SiO₂), and the eluent was evaporated to give (4-bromo-2-fluoro-6-(vinylthio)phenyl)methanol (1.6 g, 6.08 mmol, 48% yield) as a colorless oil.

[0594] 1 H NMR (400MHz, DMSO-d6) δ=7.47-7.44(m,1H),7.30-7.29(m,1H),6.77-6.70(m,1H),5.59-5.52(m,2H),5.22-5.20(m,1H),4.51-4.46(m,2H)ppm.

[0595] Step 5: Preparation of (4-bromo-2-fluoro-6-(vinylsulfinyl)phenyl)methanol

[0596] To a mixture of (4-bromo-2-fluoro-6-(vinylthio)phenyl)methanol (800 mg, 3.04 mmol) in DCM (12 mL), m-CPBA (678.98 mg, 3.34 mmol, 85% purity) was added at 0 °C, and the mixture was stirred at 25 °C for 1 hr. The reaction mixture was quenched at 0 °C by adding 20 mL of saturated aqueous Na₂SO₃ solution, diluted with 20 mL of H₂O, and extracted with EA (100 mL x 3). The combined organic layers were washed with 100 mL of brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 20 / 1-1 / 1). The fraction was concentrated under vacuum to give (4-bromo-2-fluoro-6-(vinylsulfinyl)phenyl)methanol (690 mg, 2.47 mmol, 81% yield) as a yellow solid. LCMS(ESI)m / z:[ 79 BrM+H] + =278.9;

[0597] 1H NMR (400MHz, DMSO-d6)δ=7.74-7.71(m,1H),7.61-7.60(m,1H),7.12-7.06(m,1H) ,6.05-5.92(m,2H),5.85-5.82(m,1H),4.75-4.71(m,1H),4.63-4.58(m,1H)ppm.

[0598] Step 6: Preparation of 8-bromo-6-fluoro-2,3-dihydro-5H-benzo[e][1,4]oxathiheptaine 1-oxide

[0599] NaH (186.3 mg, 4.66 mmol, 60% purity) was added to a mixture of (650 mg, 2.33 mmol) of 40 mL of DMF at 0 °C, and the mixture was stirred at 0 °C for 1 hr. The reaction solution was quenched with 50 mL of saturated NH4Cl aqueous solution and extracted with EA (50 mL x 3). The combined organic layers were washed with brine (60 mL x 3), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1-1 / 1), and the eluent was concentrated under reduced pressure to give 8-bromo-6-fluoro-2,3-dihydro-5H-benzo[e][1,4]oxathihexane 1-oxide (350 mg, 1.25 mmol, 54% yield) as a white solid.

[0600] 1 H NMR (400MHz, CDCl3) δ = 7.79 (m, 1H), 7.38-7.35 (m, 1H), 5.13 (d, J = 14.4Hz, 1H), 4.49-4.34 (m, 3H), 3.47-3.41 (m, 1H), 3.26-3.21 (m, 1H) ppm.

[0601] Step 7: Preparation of 1-oxide of 6-fluoro-2,3-dihydro-5H-benzo[e][1,4]oxathihepta-8-carboxylic acid

[0602] K₂CO₃ (475.33 mg, 3.44 mmol) was added to a mixture of 8-bromo-6-fluoro-2,3-dihydro-5H-benzo[e][1,4]oxathiheptazone 1-oxide (320 mg, 1.15 mmol), Pd(OAc)₂ (12.87 mg, 57.32 μmol), and dicyclohexyl(3-dicyclohexylphosphoniumpropyl)phosphonium; bis(tetrafluoroborate) (70.19 mg, 114.64 μmol) in DMSO (4 mL) and H₂O (0.2 mL). The mixture was then stirred at 100 °C under a CO atmosphere (15 psi) for 4 h. The mixture was diluted with water (50 mL) and extracted with EA (30 mL x 3). The aqueous layer was acidified to pH 3 with HCl solution (2 M) and extracted with EA (100 mL x 2). The combined organic phases were washed with brine, dried over Na₂SO₄, filtered, and concentrated under vacuum to give 6-fluoro-2,3-dihydro-5H-benzo[e][1,4]oxathiheptaenoic acid-8-carboxylic acid 1-oxide (210 mg, crude), a white solid, which was used directly in the next step. LCMS (ESI) m / z: [M+H] + =244.9.

[0603] Step 8: Preparation of 1,1-dioxide of 6-fluoro-2,3-dihydro-5H-benzo[e][1,4]oxathihepta-8-carboxylic acid (intermediate 5)

[0604] Add 210.00 mg, 859.81 μmol of 1-oxide of 6-fluoro-2,3-dihydro-5H-benzo[e][1,4]oxathiheptaenoic acid-8-carboxylic acid to a mixture of MeOH (4 mL) and H₂O (4 mL). (634.30 mg, 1.03 mmol), and the mixture was then stirred at 25 °C for 2 h. The mixture was diluted with water (50 mL), then extracted with EA (30 mL x 3), the combined organic layers were washed with brine (40 mL x 2), dried with Na2SO4, filtered, and concentrated under vacuum to give 1,1-dioxide of 6-fluoro-2,3-dihydro-5H-benzo[e][1,4]oxathiheptacoine-8-carboxylic acid (220 mg, crude), as a white solid, which was used directly for the next step.

[0605] Intermediates 6 and 7. (R)-2-methyl-2,3-dihydro-5H-benzo[e][1,4]oxathiheptacoine-8-carboxylic acid 1,1-dioxide and (S)-2-methyl-2,3-dihydro-5H-benzo[e][1,4]oxathiheptacoine-8-carboxylic acid 1,1-dioxide

[0606] Step 1: Preparation of 8-bromo-2-methyl-3,5-dihydro-2H-benzo[e][1,4]oxathiheptazone 1,1-dioxide

[0607] NaH (65.82 mg, 1.65 mmol, 60% purity) was added to a solution of 8-bromo-2,3-dihydro-5H-benzo[e][1,4]oxathiaheptacorene 1,1-dioxide (380 mg, 1.37 mmol, 641.03 μL) in 5 mL of DMF at 0 °C. The mixture was stirred at 0 °C for 0.5 h. Then, MeI (233.55 mg, 1.65 mmol, 102.43 μL) was slowly added at 0 °C. The mixture was stirred at 25 °C for 1.5 h. The mixture was diluted with saturated NH4Cl solution (30 mL) and extracted with EtOAc (30 mL x 2). The combined starch organic layers were dried over anhydrous Na2SO4 and concentrated to give the residue. The residue was purified by reversed-phase HPLC (0.1% FA conditions). The eluent was concentrated to remove MeCN, and then lyophilized to give 8-bromo-2-methyl-3,5-dihydro-2H-benzo[e][1,4]oxathiaheptacorine 1,1-dioxide (100 mg, 309.11 μmol, 23% yield) as a white solid.

[0608] LCMS(ESI)m / z:[M+H] + =291.0 / 292.9;

[0609] 1 HNMR (400MHz, DMSO-d6) δ = 7.99 (d, J = 2.0Hz, 1H), 7.95-7.92 (m, 1H), 7.56 (d, J = 8.0Hz, 1H), 4.87 (s, 2H), 4.27 -4.23(m,1H),4.00-3.95(m,1H),3.71-3.62(m,1H),1.14(d,J=7.2Hz,3H)ppm.

[0610] Step 2: Preparation of 1,1-dioxide of 2-methyl-3,5-dihydro-2H-benzo[e][1,4]oxathihepta-8-carboxylic acid

[0611] To a solution of 8-bromo-2-methyl-3,5-dihydro-2H-benzo[e][1,4]oxathiheptazone 1,1-dioxide (100 mg, 343.45 μmol) in DMSO (1 mL) and H₂O (30.95 mg, 1.72 mmol, 31 μL), 1,3-bis(dicyclohexylphosphino)propane bis(tetrafluoroborate) (21.03 mg, 34.35 μmol), K₂CO₃ (71.20 mg, 515.18 μmol), and Pd(OAc)₂ (7.71 mg, 34.35 μmol) were added. The flask was degassed and purged three times with CO. The mixture was stirred at 100 °C under a CO atmosphere (15 psi) for 4 h. The mixture was filtered and washed with EA (2 mL) and water (2 mL). The mixture was then diluted with water (5 mL) and extracted with EA (5 mL x 2). Discard the combined organic layers. Adjust the pH of the aqueous phase to 6 with 1N HCl aqueous solution. Then extract the aqueous phase with EA (5 mL x 2). Wash the combined organic layers with brine (5 mL x 2), dry with anhydrous Na2SO4, and concentrate to give 1,1-dioxide of 2-methyl-3,5-dihydro-2H-benzo[e][1,4]oxathiheptacosan-8-carboxylic acid (80 mg, 0.290 mol, 85% yield) as a white solid.

[0612] LCMS(ESI)m / z:[M+H] + =256.9;

[0613] 1 HNMR (400MHz, DMSO-d6) δ = 8.44 (d, J = 1.6Hz, 1H), 8.22-8.19 (m, 1H), 7.72 (d, J = 8.0Hz, 1H), 4.95 (s, 2H), 4.29 -4.25(m,1H),4.03-3.98(m,1H),3.72-3.60(m,1H),1.14(d,J=6.8Hz,3H)ppm.

[0614] Step 3: Preparation of (R)-2-methyl-3,5-dihydro-2H-benzo[e][1,4]oxathiheptacorine-8-carboxylic acid 1,1-dioxide (intermediate 6) and (S)-2-methyl-3,5-dihydro-2H-benzo[e][1,4]oxathiheptacorine-8-carboxylic acid 1,1-dioxide (intermediate 7)

[0615] Racemic 2-methyl-3,5-dihydro-2H-benzo[e][1,4]oxathiaheptaphylline-8-carboxylic acid 1,1-dioxide was separated by SFC (column: Daicel ChiralPak IG (250x30mm, 10µm); mobile phase: [0.1% NH3H2OMEOH]; B%: 30%-30%, 3.0; 85 min). The eluent was concentrated to remove most of the solvent and adjusted to pH 6 with FA. The mixture was then extracted with DCM (20 mL x 2). The combined organic layers were dried over anhydrous Na₂SO₄ and concentrated to give (R)-2-methyl-3,5-dihydro-2H-benzo[e][1,4]oxathiheptacosan-8-carboxylic acid 1,1-dioxide (35 mg, 0.136 mmol, 44% yield) as a white solid; and (S)-2-methyl-3,5-dihydro-2H-benzo[e][1,4]oxathiheptacosan-8-carboxylic acid 1,1-dioxide (40 mg, 0.156 mmol, 50.00% yield) as a white solid. Any partition stereochemistry.

[0616] (R)-2-Methyl-3,5-dihydro-2H-benzo[e][1,4]oxathiheptacoine-8-carboxylic acid 1,1-dioxide (intermediate 6): LCMS (ESI) m / z: [M+Na] + =279.1;

[0617] Chiral SFC: IG-3_5CM_MEOH(DEA)_5_40_3ML_T35.M; Rt=1.729min;

[0618] (S)-2-Methyl-3,5-dihydro-2H-benzo[e][1,4]oxathiheptaenoic acid-8-carboxylic acid 1,1-dioxide (intermediate 7): LCMS (ESI) m / z: [M+Na] + =279.1;

[0619] Chiral SFC: IG-3_5CM_MEOH(DEA)_5_40_3ML_T35.M; Rt=1.897min.

[0620] Intermediate 8,6,7,8,9-tetrahydrothiopheno[3,2-b]pyridine-3-carboxylic acid 5,5-dioxide

[0621]

[0622] Step 1: Preparation of 2-bromo-5-chloro-pyridine-3-thiol

[0623] Na₂S (482.14 mg, 6.18 mmol, 1 eq) was added in a single batch to a mixture of 2-bromo-5-chloro-3-fluoro-pyridine (1.3 g, 6.18 mmol, 1 eq) in DMF (20 mL) under a nitrogen atmosphere at 25 °C. The mixture was stirred at 25 °C for 12 h. The mixture was then poured into a 100 mL container. An aqueous solution of HCl (2 M) was added to the mixture to adjust the pH to 3. The aqueous phase was extracted with ethyl acetate (50 mL x 2). The combined organic phases were washed with brine (50 mL x 1), dried over anhydrous Na₂SO₄, filtered, and concentrated under vacuum to give 2-bromo-5-chloro-pyridine-3-thiol (1.2 g, 5.35 mmol, 86.52% yield) as a yellow solid.

[0624] LCMS(ESI)m / z:[ 79 BrM+H] + =225.8.

[0625] 1 H NMR (400MHz, DMSO-d6) δ = 8.33-8.23 (m, 1H), 8.17-8.08 (m, 1H) ppm.

[0626] Step 2: Preparation of 2-bromo-3-(but-3-en-1-ylthio)-5-chloropyridine

[0627] PPh3 (2.10 g, 8.02 mmol) was added to a mixture of 2-bromo-5-chloro-pyridin-3-thiol (1.2 g, 5.35 mmol) and but-3-en-1-ol (385.41 mg, 5.35 mmol, 459.91 μL) in THF (10 mL), followed by dropwise addition of DEAD (1.40 g, 8.02 mmol, 1.46 mL) at 0 °C under a nitrogen atmosphere. The mixture was stirred at 25 °C for 12 h. The mixture was poured into water (50 mL) and extracted with ethyl acetate (30 mL x 2). The combined organic phases were washed with brine (30 mL x 1), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum to give the residue. The residue was purified by silica gel chromatography (petroleum ether / ethyl acetate = 10 / 1). The eluent was concentrated to give 2-bromo-3-(but-3-en-1-ylthio)-5-chloropyridine (1.2 g, 4.31 mmol, 81% yield) as a yellow oil. LCMS (ESI) m / z: [ 79 BrM+H] + =277.9.

[0628] 1H NMR (400MHz, CDCl3) δ=8.05-7.99(m,1H),7.34-7.28(m,1H),5.88-5.73(m,1H),5.16-5.01(m,2H),2.99-2.86(m,2H),2.48-2.35(m,2H)ppm.

[0629] Step 3: Preparation of 3-(but-3-en-1-ylthio)-5-chloro-2-vinylpyridine

[0630] A mixture of 2-bromo-3-(but-3-en-1-ylthio)-5-chloropyridine (860 mg, 3.09 mmol), potassium vinyltrifluoroborate (1.24 g, 9.26 mmol), Pd(dtbpf)Cl2 (201.19 mg, 308.69 μmol), and K3PO4 (1.97 g, 9.26 mmol) in dioxane (12 mL) and H2O (3 mL) was stirred at 80 °C under a nitrogen atmosphere for 1 hr. The mixture was poured into H2O (100 mL) and extracted with EA (30 mL x 3). The combined organic layers were washed with brine (20 mL x 2), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by silica gel chromatography (PE–PE / EA = 20 / 1). The eluent was concentrated under reduced pressure to give 3-(but-3-en-1-ylthio)-5-chloro-2-vinylpyridine (510 mg, 2.26 mmol, 73.% yield) as a yellow oil. LCMS (ESI) m / z: [M+H] + =226.0.

[0631] 1 H NMR (400MHz, CDCl3) δ = 8.36 (d, J = 2.4Hz, 1H), 7.60 (d, J = 2.4Hz, 1H), 7.26-7.19 (m, 1H), 6.41-6.36 (m, 1 H),5.89-5.81(m,1H),5.57-5.53(m,1H),5.20-5.07(m,2H),2.97-2.93(m,2H),2.44-2.36(m,2H)ppm.

[0632] Step 4: Preparation of 3-chloro-6,7-dihydrothiopheno[3,2-b]pyridine

[0633] A mixture of 3-(but-3-en-1-ylthio)-5-chloro-2-vinylpyridine (250 mg, 1.11 mmol) and benzylidene-[1,3-bis(2,4,6-trimethylphenyl)imidazolidine-2-ylidene]-dichloro-ruthenium; tricyclohexylphosphine (Grubbs II) (94.0 mg, 0.111 mol) in DCM (12 mL) was stirred at 25 °C under N2 atmosphere for 16 h. The solution was concentrated under vacuum. The residue was purified by silica gel chromatography (PE–PE / EA = 20 / 1). The eluent was concentrated under reduced pressure to give 3-chloro-6,7-dihydrothieno[3,2-b]pyridine (110 mg, 556.44 μmol, 50% yield) as a yellow oil. LCMS (ESI) m / z: [M+H] + =198.0.

[0634] 1 H NMR (400MHz, CDCl3) δ = 8.40 (d, J = 2.0Hz, 1H), 7.71 (d, J = 1.6Hz, 1H), 6.81-6.71 (m, 1H), 6.32-6.26 (m, 1H), 3.10-3.05 (m, 2H), 2.88-2.81 (m, 2H).

[0635] Step 5: Preparation of 6,7-dihydrothiopheno[3,2-b]pyridine-3-carboxylic acid

[0636] A mixture of 3-chloro-6,7-dihydrothieno[3,2-b]pyridine (50 mg, 0.253 mol), K₂CO₃ (52.44 mg, 0.379 mol), Pd(OAc)₂ (2.84 mg, 12.65 μmol), 1,3-bis(dicyclohexylphosphino)propanebis(tetrafluoroborate) (15.49 mg, 25.29 μmol), and H₂O (100 μL) in DMSO (1 mL) was stirred at 100 °C under a CO atmosphere (15 psi) for 4 h. The mixture was then poured into H₂O (10 mL) and extracted with EA (10 mL x 2). The organic phase was discarded. The aqueous phase was acidified to pH 3 with HCl (1 M) and extracted with EA (10 mL x 3). The combined organic layers were washed with brine (10 mL x 2), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to give 6,7-dihydrothieno[3,2-b]pyridine-3-carboxylic acid (28 mg, 135.10 μmol, 53.4% ​​yield) as a white solid. LCMS (ESI) m / z: [M+H] + =207.9.

[0637] 1H NMR (400MHz, CDCl3) δ = 8.36 (d, J = 2.4Hz, 1H), 7.60 (d, J = 2.4Hz, 1H), 7.26-7.19 (m, 1H), 6.41-6.36 (m, 1 H),5.89-5.81(m,1H),5.57-5.53(m,1H),5.20-5.07(m,2H),2.97-2.93(m,2H),2.44-2.36(m,2H)ppm.

[0638] Step 6: Preparation of 6,7,8,9-tetrahydrothiopheno[3,2-b]pyridine-3-carboxylic acid

[0639] Pd / C (wet, 50 mg, 10% purity) was added to a mixture of 6,7-dihydrothieno[3,2-b]pyridine-3-carboxylic acid (28 mg, 135.10 μmol) in MeOH (5 mL) at 25 °C. The mixture was purged three times with H2 and stirred for 30 min at 25 °C under H2 atmosphere (15 psi). The mixture was filtered, and the filtrate was concentrated under reduced pressure to give 6,7,8,9-tetrahydrothieno[3,2-b]pyridine-3-carboxylic acid (23 mg, 109.91 μmol, 81.2% yield) as a white solid. LCMS (ESI) m / z: [M+H] + =210.0.

[0640] 1 H NMR (400MHz, DMSO-d6) δ = 8.81 (d, J = 2.0Hz, 1H), 8.20 (d, J = 2.0Hz, 1H), 3.23-3.17 (m, 2H), 2.88-2.78 (m, 2H), 2.11-1.96 (m, 2H), 1.76-1.62 (m, 2H)ppm.

[0641] Step 7: Preparation of 5,5-dioxo-6,7,8,9-tetrahydrothieno[3,2-b]pyridine-3-carboxylic acid (intermediate 8)

[0642] Add 23 mg, 109.91 μmol of 6,7,8,9-tetrahydrothiopheno[3,2-b]pyridine-3-carboxylic acid to a mixture of MeOH (1 mL) and H₂O (1 mL) at 25 °C. (67.57 mg, 109.9 μmol). The mixture was stirred at 25 °C for 4 h. The mixture was quenched with saturated Na₂SO₃ (20 mL), acidified with HCl (1 M) to pH 2, and extracted with EA (20 mL x 2). The combined organic layers were washed with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to give 5,5-dioxo-6,7,8,9-tetrahydrothieno[3,2-b]pyridine-3-carboxylic acid (18 mg, 74.61 μmol, 68% yield) as a white solid.

[0643] LCMS(ESI)m / z:[M+H] + =241.9;

[0644] 1 H NMR (400MHz, DMSO-d6) δ = 9.14 (d, J = 2.0Hz, 1H), 8.55 (d, J = 2.1Hz, 1H), 3.53-3.51 (m, 2H), 3.17 (br d, J = 5.2Hz, 2H), 2.19-2.13 (m, 2H), 1.82 (br d,J=3.2Hz,2H)ppm.

[0645] Intermediate 9: 4,5-dihydro-2H-benzo[d][1,3]oxathihepta-8-carboxylic acid 1,1-dioxide

[0646] Step 1: Preparation of (6-bromobenzo[b]thiophene-2-yl)boronic acid

[0647] LDA (2M, 22.53mL) was added dropwise to a mixture of 6-bromobenzo[b]thiophene (8g, 37.54mmol) in THF (80mL) at -70°C under a nitrogen atmosphere. The mixture was stirred at -70°C for 1 hour. Then, triisopropyl borate (8.47g, 45.05mmol, 10.36mL) was added to the mixture at -70°C, and the mixture was stirred for 1 hour. H₂SO₄ (7.36g, 75.08mmol, 4.00mL) was added to the mixture at -70°C, and the mixture was stirred at 25°C for 1 hour. The mixture was poured into water (300mL) and extracted with ethyl acetate (200mL x 2). The combined organic phases were washed with brine (200mL x 1), dried over anhydrous Na₂SO₄, filtered, and concentrated under vacuum. The residue was ground with PE / MTBE = 10 / 1 (50mL). The suspension was filtered. The filter cake was dried in a pump to give (6-bromobenzo[b]thiophene-2-yl)boronic acid (7.3 g, 28.41 mmol, 76% yield) as a pale yellow solid. 1H NMR (400MHz, DMSO-d6) δ=8.58-8.53(m,2H),8.28-8.24(m,1H),7.96-7.93(m,1H),7.88-7.84(m,1H),7.54-7.46(m,1H)ppm.

[0648] Step 2: Preparation of 6-bromobenzo[b]thiophene-2(3H)-one

[0649] H₂O₂ (38.35 g, 338.24 mmol, 32.50 mL) was added dropwise to a mixture of (6-bromobenzo[b]thiophene-2-yl)boric acid (6.5 g, 25.30 mmol) in EtOH (78 mL) under a nitrogen atmosphere at 25 °C. The mixture was stirred at 25 °C for 1 hr. The mixture was filtered. The filter cake was washed with H₂O (50 mL) and dried under vacuum to give 6-bromobenzo[b]thiophene-2(3H)-one (4.2 g, 18.33 mmol, 72% yield) as a brown solid. LCMS (ESI) m / z: [M+H] + =214.8,216.9.

[0650] 1 H NMR (400MHz, CDCl3) δ = 7.53-7.47 (m, 1H), 7.38-7.32 (m, 1H), 7.16 (d, J = 8.0Hz, 1H), 4.06-3.84 (m, 2H) ppm.

[0651] Step 3: Preparation of 2-(4-bromo-2-mercaptophenyl)ethanol-1-ol

[0652] In a nitrogen atmosphere at 25 °C, NaBH4 (3.47 g, 91.67 mmol) was added fractionally to a mixture of 6-bromobenzo[b]thiophene-2(3H)-one (4.2 g, 18.33 mmol) in EtOH (67 mL). The mixture was stirred at 80 °C for 30 min. The mixture was then cooled to 25 °C. An aqueous solution of HCl (1 M) was slowly added to the mixture to adjust the pH to 2. The mixture was poured into water (200 mL) and extracted with ethyl acetate (100 mL x 2). The combined organic phases were washed with brine (100 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (petroleum ether / ethyl acetate = 2 / 1). The eluent was concentrated to give 2-(4-bromo-2-mercaptophenyl)ethyl-1-ol (3.6 g, 15.44 mmol, 84% yield) as a yellow oil. 1H NMR (400MHz, DMSO-d6) δ = 7.61 (d, J = 2.0Hz, 1H), 7.24-7.22 (m, 1H), 7.13 (d, J = 8. 2Hz,1H),5.58(s,1H),4.97-4.49(m,1H),3.59-3.57(m,2H),2.71-2.69(m,2H).

[0653] Step 4: Preparation of 8-bromo-4,5-dihydrobenzo[d][1,3]oxathiaheptaine

[0654] NaH (257.37 mg, 6.43 mmol) was added fractionally to a mixture of 2-(4-bromo-2-mercaptophenyl)ethyl-1-ol (500 mg, 2.14 mmol) in DMF (50 mL) at 0 °C under a nitrogen atmosphere. The mixture was stirred at 25 °C for 30 min. Then, chloro(iodine)methane (416.13 mg, 2.36 mmol, 171 μL) in DMF (1 mL) was added dropwise to the mixture at 0 °C under a nitrogen atmosphere. The mixture was stirred at 25 °C for 1.5 h. The mixture was poured into saturated NH4Cl (10 mL) and extracted with ethyl acetate (10 mL x 2). The combined organic phases were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (petroleum ether / ethyl acetate = 10 / 1). The eluent was concentrated to give 8-bromo-4,5-dihydrobenzo[d][1,3]oxathiaheptacoine (50 mg, 0.189 mmol, 9% yield) as a yellow oil.

[0655] LCMS(ESI)m / z:[M+H] + =246.2,248.0.

[0656] 1 H NMR (400MHz, DMSO-d6) δ=7.67-7.60(m,1H),7.47-7.38(m,1H),7.31-7.22(m,1H),5.00-4.86(m,2H),3.81-3.67(m,2H),3.12-3.09(m,2H)ppm.

[0657] Step 5: Preparation of 4,5-dihydrobenzo[d][1,3]oxathiaheptaphylline-8-carboxylic acid

[0658] A solution of 8-bromo-4,5-dihydrobenzo[d][1,3]oxathiaheptaecamyne (50 mg, 203.97 μmol), Pd(OAc)₂ (4.58 mg, 20.40 μmol), 1,3-bis(dicyclohexylphosphino)propanebis(tetrafluoroborate) (24.98 mg, 40.79 μmol), and K₂CO₃ (56.38 mg, 0.408 mmol) in DMSO (2 mL) and H₂O (0.2 mL) was degassed under vacuum and purged several times with CO. The mixture was stirred at 100 °C under a CO atmosphere (15 psi) for 2 h. The mixture was poured into water (20 mL) and extracted with ethyl acetate (10 mL x 2). The organic layer was discarded. An aqueous solution of HCl (1 M) was added to the aqueous phase to adjust the pH to 3. The mixture was then extracted with ethyl acetate (10 mL x 2). The combined organic phases were washed with brine (10 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under vacuum to give 4,5-dihydrobenzo[d][1,3]oxathiaheptacoine-8-carboxylic acid (40 mg, 190.25 μmol, 93% yield) as a yellow solid. LCMS (ESI) m / z: [M+H] + =211.1.

[0659] Step 6: Preparation of 1,1-dioxide of 4,5-dihydro-2H-benzo[d][1,3]oxathiheptaenoic acid-8-carboxylic acid (intermediate 9)

[0660] mCPBA (48.28 mg, 237.81 μmol, 85% purity) was added fractionally to a mixture of 4,5-dihydrobenzo[d][1,3]oxathiaheptaphylline-8-carboxylic acid (20 mg, 95.13 μmol) in DCM (1 mL) at 25 °C under a N2 atmosphere. The mixture was stirred at 25 °C for 12 h. The mixture was filtered, and the filtrate was concentrated. The residue was purified directly by reverse-phase column chromatography (FA). The eluent was concentrated to remove MeCN. The aqueous phase was lyophilized to give 1,1-dioxide of 4,5-dihydrobenzo[d][1,3]oxathiaheptaphylline-8-carboxylic acid (20 mg, 82.56 μmol, 86.79% yield) as a white solid. LCMS (ESI) m / z: [M+H2O] + =260.0;

[0661] 1 H NMR (400MHz, DMSO-d6) δ = 8.38 (d, J = 1.6Hz, 1H), 8.16-8.14 (m, 1H), 7.62 (d, J = 7.8Hz, 1H), 4.99 (s, 2H), 4.01-4.00 (m, 2H), 3.42 (s, 2H) ppm.

[0662] Intermediate 10: 4,5-dihydro-2H-benzo[d][1,3]oxathihepta-8-carboxylic acid 1,1-dioxide

[0663] Step 1: Preparation of 4-bromo-2-[(4-methoxyphenyl)methylthioalkyl]benzylnitrile

[0664] Cs₂CO₃ (16.29 g, 50.00 mmol) was added to a solution of 4-bromo-2-fluorobenzyl nitrile (10 g, 50.00 mmol) and (4-methoxyphenyl)methanethiol (7.71 g, 50.00 mmol) in DMF (100 mL), and the mixture was stirred at 60 °C for 2 h. The reaction mixture was poured into water (1000 mL), and the solution was extracted with EA (1000 mL x 3). The combined organic layers were washed with brine (500 mL), dried over Na₂SO₄, filtered, and concentrated to give 4-bromo-2-[(4-methoxyphenyl)methylthioalkyl]benzyl nitrile (13 g, crude) as a white solid.

[0665] Step 2: Preparation of [4-bromo-2-[(4-methoxyphenyl)methylthioalkyl]phenyl]methylamine

[0666] In a nitrogen atmosphere at 0°C, LiAlH4 (1.62 g, 42.78 mmol) was added to a solution of 13 g (38.90 mmol) of 4-bromo-2-[(4-methoxyphenyl)methylthioalkyl]benzylnitrile in 150 mL of THF. The mixture was stirred at 0°C for 1 hour. Water (1.62 g) and 15% NaOH solution (2.5 mL) were added to the mixture. The solution was then poured into EA (500 mL), filtered, and the filtrate was concentrated to obtain [4-bromo-2-[(4-methoxyphenyl)methylthioalkyl]phenyl]methylamine (13 g, crude), which is a yellow oily substance.

[0667] 1H NMR (400MHz, DMSO-d6) δ=7.48-7.47(m,1H),7.21-7.20(m,1H),7.19-7.18(m,3H),6.85-6.82(m,2H),4.08(s,2H),3.80-3.79(m,5H)ppm.

[0668] Step 3: Preparation of [2-[[2-(aminomethyl)-5-bromo-phenyl]dithioalkyl]-4-bromo-phenyl]methylamine

[0669] The mixture of [4-bromo-2-[(4-methoxyphenyl)methylthioalkyl]phenyl]methylamine (13 g, 38.43 mmol) in TFA (130 mL) was stirred at 60 °C for 16 h. The reaction mixture was concentrated to give the residue. The residue was purified by reversed-phase HPLC (0.1% FA conditions). The solution was lyophilized to give [aminomethyl]-5-bromo-phenyl]dithioalkyl]-4-bromo-phenyl]methylamine (3.5 g, 7.20 mmol, 19% yield) as a white solid.

[0670] LCMS(ESI)m / z:[ 79 BrM+H] + =434.8.

[0671] 1H NMR (400MHz, DMSO-d6) δ = 8.35 (br s, 3H), 7.55 (s, 2H), 7.50-7.37 (m, 1H), 4.05 (s, 2H) ppm.

[0672] Step 4: Preparation of 8-bromo-4,5-dihydro-1,4-benzothiazepin-3-one

[0673] To a solution of [aminomethyl]-5-bromo-phenyl]dithio]-4-bromo-phenyl]methylamine (1 g, 2.30 mmol) in THF (15 mL), NaBH4 (261.37 mg, 6.91 mmol) was added, and the mixture was stirred at 30 °C for 2 h. Then, TEA (11.52 mmol, 1.60 mL) and 2-chloroacetyl chloride (312.13 mg, 2.76 mmol) were added to the solution, and the mixture was stirred at 30 °C for 3 h. The reaction mixture was poured into water (100 mL) and extracted with EA (100 mL x 3). The combined organic layers were washed with brine (200 mL), dried over Na2SO4, filtered, and concentrated to obtain the residue. The residue (SiO2, petroleum ether / ethyl acetate = 10:1-0:1) was purified by column chromatography, and the solution was concentrated to give 8-bromo-4,5-dihydro-1,4-benzothiazohepta-3-one (300 mg, 871.29 μmol, 38% yield) as a white solid.

[0674] LCMS(ESI)m / z:[ 79 BrM+H] + =260.0.

[0675] 1H NMR (400MHz, DMSO-d6) δ = 7.37 (d, J = 2.0Hz, 1H), 7.24–7.22 (m, 1H), 7.07 (d, J = 8.0Hz, 1H), 4.45 (s, 2H), 3.89 (s, 2H) ppm.

[0676] Step 5: Preparation of 3-oxo-4,5-dihydro-1,4-benzothiazoheptaenoic acid-8-carboxylic acid (intermediate 10)

[0677] To a solution of 280 mg, 1.08 mmol, of 8-bromo-4,5-dihydro-1,4-benzothiazoheptaeno-3-one in DMSO (5 mL), dicyclohexyl(3-dicyclohexylphosphoniumpropyl)phosphonium; bis(tetrafluoroborate) (66.41 mg, 108.47 μmol), K₂CO₃ (224.88 mg, 1.63 mmol), Pd(OAc)₂ (24.35 mg, 108.47 μmol), and H₂O (3.91 mg, 216.94 μmol) were added. The mixture was stirred at 100 °C for 2 h under a CO atmosphere (15 psi). The reaction mixture was filtered, and the solution was extracted with MTBE (10 mL), discarding the organic layer. The aqueous phase was then adjusted to pH 2 with 1N HCl, and the solution was extracted with EA (50 mL x 5). The combined organic layers were washed with brine (100 mL), dried with Na2SO4, filtered, and concentrated to give 3-oxo-4,5-dihydro-1,4-benzothiazohepta-8-carboxylic acid (120 mg, 0.487 mmol, 45% yield) as a white solid.

[0678] LCMS(ESI)m / z:[M+H] + =224.1.

[0679] 1 H NMR (400MHz, DMSO-d6) δ=13.09–13.06(m,1H),8.18(t,J=6.4Hz,1H),7.64(d,J=1.6Hz ,1H),7.60-7.57(m,1H),7.29(d,J=8.0Hz,1H),4.45(d,J=6.4Hz,2H),3.91(s,2H)ppm.

[0680] Step 6: Preparation of 1,1,3-trioxo-4,5-dihydro-1λ6,4-benzothiazoheptan-8-carboxylic acid

[0681] Oxone (275.37 mg, 447.93 μmol) was added to a solution of 3-oxo-4,5-dihydro-1,4-benzothiazohepta-8-carboxylic acid (50 mg, 223.97 μmol) in MeOH (0.5 mL) and H₂O (0.5 mL). The mixture was stirred at 30 °C for 2 h. The reaction mixture was poured into MeOH (5 mL), filtered, and the filtrate was concentrated to give 1,1,3-trioxo-4,5-dihydro-1λ6,4-benzothiazohepta-8-carboxylic acid (57 mg, 223.31 μmol, 99.71% yield) as a white solid.

[0682] Intermediate 11,4-(2-methoxyethyl)-3-methylsulfonyl-benzoic acid

[0683]

[0684] Step 1: Preparation of 2-(4-chloro-2-methylthioalkyl-phenyl)acetic acid

[0685] A mixture of 2-(2-bromo-4-chlorophenyl)acetic acid (1 g, 4.01 mmol), CuI (763.36 mg, 4.01 mmol), and DABCO (899.20 mg, 8.02 mmol, 881.57 μL) in DMSO (10 mL) was stirred at 145 °C under a N2 atmosphere for 12 h. The reaction mixture was diluted with 1 N HCl (300 mL) and filtered. The filtrate was extracted with DCM (300 mL x 2). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated to give the residue. The residue was purified by column chromatography (petroleum ether / ethyl acetate = 1 / 0-0 / 1). The eluent was concentrated to give 2-(4-chloro-2-methylthioalkyl-phenyl)acetic acid (1.5 g, crude), a yellow solid, which was used directly in the next step.

[0686] Step 2: Preparation of 2-(4-chloro-2-methylsulfonyl-phenyl)acetic acid

[0687] Oxone (4.26 g, 6.92 mmol) in H2O (3 mL) was added to a solution of 2-(4-chloro-2-methylthiophenyl)acetic acid (500 mg, 2.31 mmol) in MeOH (3 mL) and H2O (3 mL) at 0 °C. The mixture was stirred at 25 °C for 12 h. The reaction mixture was diluted with saturated Na2SO3 (100 mL), stirred for 10 min, and then extracted with DCM (100 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated to give the residue. The residue was purified by reverse-phase (0.1% FA). The eluent was concentrated to give 2-(4-chloro-2-methylsulfonylphenyl)acetic acid (200 mg, 0.804 mol, 35% yield) as a white solid. LCMS (ESI) m / z: [M+H] + =248.9.

[0688] 1 H NMR (400MHz, DMSO-d6) δ = 12.62-12.54 (m, 1H), 7.91 (d, J = 2.0Hz, 1H), 7.78-7.76 (m, 1H), 7.55 (d, J = 8.0Hz, 1H), 4.05 (s, 2H), 3.26 (s, 3H) ppm.

[0689] Step 3: Preparation of 2-(4-chloro-2-methylsulfonyl-phenyl)ethanol.

[0690] BH3-Me2 was added to a solution of 2-(4-chloro-2-methylsulfonyl-phenyl)acetic acid (200 mg, 804.24 μmol) in THF (4 mL) at 0 °C. S The mixture (10M, 402.12 μL) was stirred at 25 °C for 2 h. The reaction mixture was diluted with 1N HCl (10 mL) and extracted with DCM (10 mL). The organic layer was dried over Na₂SO₄-free water, filtered, and concentrated to give the residue. The residue was purified by reverse-phase chromatography (0.1% FA). The eluent was concentrated to give 2-(4-chloro-2-methylsulfonyl-phenyl)ethanol (180 mg, 766.94 μmol, 96% yield) as a colorless oil. LCMS (ESI) m / z: [M+H] + =235.0.

[0691] 1 H NMR (400MHz, CDCl3) δ = 8.06 (d, J = 2.4Hz, 1H), 7.58-7.55 (m, 1H), 7.42 (d, J = 8.0Hz, 1H), 3.97-3.94 (m, 2H), 3.28-3.25 (m, 2H), 3.15 (s, 3H) ppm.

[0692] Step 4: Preparation of 4-chloro-1-(2-methoxyethyl)-2-methylsulfonyl-benzene

[0693] Ag₂O (236.97 mg, 1.02 mmol) and MeI (241.91 mg, 1.70 mmol, 10⁶ μL) were added to a solution of 2-(4-chloro-2-methylsulfonyl-phenyl)ethanol (80 mg, 0.341 mmol) in DCM (1 mL). The mixture was stirred at 30 °C for 12 h. The reaction mixture was diluted with H₂O (10 mL) and extracted with DCM (10 mL x 2). The combined organic layers were dried over anhydrous Na₂SO₄, filtered, concentrated, and the residue was purified by reverse-phase (0.1% FA). The eluent was concentrated to give 4-chloro-1-(2-methoxyethyl)-2-methylsulfonyl-phenyl (60 mg, 0.241 mmol, 71% yield) as a yellow solid.

[0694] LCMS(ESI)m / z:[M+H] + =248.9.

[0695] 1 H NMR (400MHz, CDCl3) δ = 8.06 (d, J = 2.4Hz, 1H), 7.55-7.52 (m, 1H), 7.42 (d, J = 8.4Hz, 1H), 3.70-3.67 (m, 2H), 3.33-3.29 (m, 5H), 3.15 (s, 3H) ppm.

[0696] Step 5: Preparation of 4-(2-methoxyethyl)-3-methylsulfonyl-benzoic acid (intermediate 11)

[0697] A mixture of 4-chloro-1-(2-methoxyethyl)-2-methylsulfonylbenzene (60 mg, 0.241 mmol), K₂CO₃ (50.0 mg, 0.362 mmol), dicyclohexyl(3-dicyclohexylphosphoniumpropyl)phosphonium; bis(tetrafluoroborate) (14.77 mg, 24.12 μmol), and Pd(OAc)₂ (2.71 mg, 12.06 μmol) in DMSO (1 mL) and H₂O (0.2 mL) was degassed and purged three times with CO. The mixture was stirred at 100 °C under a CO atmosphere (15 psi) for 3 h. The reaction mixture was diluted with MeOH (10 mL), filtered, and the filtrate was concentrated to give the residue. The residue was purified by reverse-phase (0.1% FA). The eluent was concentrated to remove ACN, and then lyophilized to give 4-(2-methoxyethyl)-3-methylsulfonylbenzoic acid (50 mg, 0.194 mmol, 80% yield) as a white solid. LCMS (ESI) m / z: [M+H] + =259.0.

[0698] 1 H NMR (400MHz, CDCl3) δ = 8.78 (d, J = 1.6Hz, 1H), 8.28-8.26 (m, 1H), 7.61 (d, J = 8. 4Hz,1H),3.77-3.74(m,2H),3.45-3.42(m,2H),3.33(s,3H),3.19(s,3H)ppm.

[0699] Intermediate 12,4-(2-methoxyethyl)-3-methylsulfonyl-benzoic acid

[0700]

[0701] Step 1: Preparation of methyl 3-[allyl(tert-butoxycarbonyl)aminosulfonyl]-4-vinylbenzoate

[0702] TEA (863.24 mg, 8.53 mmol, 1.19 mL) and Boc₂O (1.86 g, 8.53 mmol, 1.96 mL) were added to a solution of methyl 3-(allylaminosulfonyl)-4-vinylbenzoate (1.2 g, 4.27 mmol) (prepared according to the method in FG-A4366) and DMAP (52.11 mg, 426.55 μmol) in DCM (20 mL) at 0 °C. The mixture was stirred at 20 °C for 2 h. It was poured into water (60 mL) and extracted with DCM (40 mL x 3). The combined organic layers were washed with brine (40 mL), dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel rapid chromatography (…). 40g Rapid column chromatography with a silica gel column, eluent gradient of 0–50% ethyl acetate / petroleum ether @ 50 mL / min. Vacuum concentration fractionation yielded methyl 3-[allyl(tert-butoxycarbonyl)aminosulfonyl]-4-vinylbenzoate (1.5 g, 3.93 mmol, 92% yield) as a yellow oil.

[0703] 1 H NMR (400MHz, DMSO-d6) δ = 8.50 (d, J = 2.0Hz, 1H), 8.34-8.15 (m, 1H), 7.92 (d, J = 8.4Hz, 1H), 7.23-7.00 (m, 1H), 6.01-5.86(m,2H),5.75-5.61(m,1H),5.39-5.14(m,2H),4.38(d,J=4.8Hz,2H),3.91(s,3H),1.13(s,9H)ppm.

[0704] Step 2: Preparation of 1,1-dioxide of benzo[f][1,2]thiazoheptan-2,8(3H)-dicarboxylic acid 2-(tert-butyl) ester 8-methyl ester

[0705] The mixture of methyl 3-[allyl(tert-butoxycarbonyl)aminosulfonyl]-4-vinylbenzoate (1.5 g, 3.93 mmol) and benzylidene-[1,3-bis(2,4,6-trimethylphenyl)imidazolidine-2-ylidene]-dichloro-ruthenium; tricyclohexylphosphine (333.85 mg, 393.24 μmol) in DCM (80 mL) was degassed and purged three times with N2. The mixture was stirred at 25 °C under N2 atmosphere for 2 h. The concentration was used to remove DCM. The residue was purified by silica gel rapid chromatography. 40g Silica gel rapid column chromatography, eluent with a 0–50% ethyl acetate / petroleum ether gradient @ 50 mL / min). Vacuum concentration fractionation yielded benzo[f][1,2]thiazohepta-2,8(3H)-dicarboxylic acid 2-(tert-butyl) ester 8-methyl ester 1,1-dioxide (1.1 g, 2.77 mmol, 70% yield), as a yellow solid. LCMS (ESI) m / z: [Br 79 M+H] + =298.0

[0706] 1H NMR (400MHz, DMSO-d6) δ = 8.43 (d, J = 1.6Hz, 1H), 8.32-8.17 (m, 1H), 7.82 (d, J = 8.0Hz, 1H), 6 .75(d,J=12.8Hz,1H),6.39-6.18(m,1H),4.95-4.57(m,2H),3.92(s,3H),1.11(s,9H)ppm.

[0707] Step 3: Preparation of 1,1-dioxide of 8-methyl 2-(tert-butyl)-2,8(3H)-dicarboxylic acid ester 4,5-dihydrobenzo[f][1,2]thiazohepta-2,8(3H)-dicarboxylic acid ester 2-(tert-butyl) ester

[0708] A mixture of benzo[f][1,2]thiaheptaphylline-2,8(3H)-dicarboxylic acid 2-(tert-butyl) ester 8-methyl ester 1,1-dioxide (500 mg, 1.41 mmol) and Pd / C (50 mg, 10% purity) in MeOH (10 mL) was degassed and purged three times with H2. The mixture was stirred at 20 °C under an H2 atmosphere for 16 h. After filtration and concentration, 4,5-dihydrobenzo[f][1,2]thiaheptaphylline-2,8(3H)-dicarboxylic acid 2-(tert-butyl) ester 8-methyl ester 1,1-dioxide (4.1 g, 12.27 mmol, 96% yield) was obtained as a yellow oil.

[0709] LCMS(ESI)m / z:[Br 79 M+H] + =300.0.

[0710] 1 H NMR (400MHz, DMSO-d6) δ = 8.37 (d, J = 2.0Hz, 1H), 8.23-8.11 (m, 1H), 7.66 (d, J = 8.0Hz, 1H) ,4.17-4.06(m,2H),3.90(s,3H),3.32-3.14(m,2H),1.90-1.53(m,2H),1.22(s,9H)ppm.

[0711] Step 4: Preparation of 2-tert-butoxycarbonyl-1,1-dioxo-4,5-dihydro-3H-1λ6,2-benzothiazoheptan-8-carboxylic acid

[0712] LiOH·H2O (118.06 mg, 2.81 mmol) was added to a solution of 2-(tert-butyl) 8-methyl 4,5-dihydrobenzo[f][1,2]thiaheptacoine-2,8(3H)-dicarboxylic acid ester 1,1-dioxide (250 mg, 0.703 mmol) in THF (2.5 mL) and H2O (2.5 mL). The mixture was stirred at 25 °C for 2 h. The pH was adjusted to 5 with aqueous HCl (1 M), and the mixture was extracted with EA (40 mL x 3). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated to give 2-tert-butoxycarbonyl-1,1-dioxo-4,5-dihydro-3H-1λ6,2-benzothiaheptacoine-8-carboxylic acid (190 mg, 0.473 mmol, 67% yield) as a white solid. LCMS(ESI)m / z:[M+H] + =285.9.

[0713] Step 5: Preparation of 1,1-dioxo-2,3,4,5-tetrahydro-1λ6,2-benzothiazoheptan-8-carboxylic acid (intermediate 12)

[0714] A mixture of 2-tert-butoxycarbonyl-1,1-dioxo-4,5-dihydro-3H-1λ6,2-benzothiazohepta-8-carboxylic acid (180 mg, 0.527 mmol) in HCl / dioxane (4 M, 3 mL) was stirred at 25 °C for 2 h. The mixture was concentrated to remove dioxane, yielding 1,1-dioxo-2,3,4,5-tetrahydro-1λ6,2-benzothiazohepta-8-carboxylic acid (130 mg, 0.468 mmol, 89% yield, HCl) as a yellow solid.

[0715] 1 H NMR (400MHz, DMSO-d6) δ = 8.31 (d, J = 1.6Hz, 1H), 8.05-8.00 (m, 1H), 7.57-7.52 (m, 2H), 3.66 (br s, 2H), 3.22 (br d,J=3.2Hz,2H),1.91-1.77(m,1H),1.70(br s,2H)ppm.

[0716] Intermediate 13,2-methyl-1,1-dioxo-4,5-dihydro-3H-1λ6,2-benzothiazoheptan-8-carboxylic acid

[0717] Step 1: Preparation of 4-bromo-3-chlorosulfonylbenzoic acid

[0718] A mixture of 4-bromobenzoic acid (10 g, 49.75 mmol) and HSO3Cl (86.95 g, 0.746 mol, 49.7 mL) was stirred at 100 °C for 16 h. The reaction mixture was then stirred at 120 °C for another 16 h. The mixture was poured into ice water (400 mL). A precipitate formed, and the mixture was filtered. The filter cake was dried under vacuum to give 4-bromo-3-chlorosulfonylbenzoic acid (11 g, 36.72 mmol, 73% yield) as a gray solid.

[0719] LCMS(ESI)m / z:[Br 79 M+H] + =300.0.

[0720] 1 H NMR (400MHz, DMSO-d6) δ = 13.96 (br s, 1H), 8.46 (d, J = 1.6Hz, 1H), 7.79-7.60 (m, 2H) ppm.

[0721] Step 2: Preparation of methyl 4-bromo-3-chlorosulfonylbenzoate

[0722] A mixture of 4-bromo-3-chlorosulfonylbenzoic acid (11 g, 36.72 mmol) and SOCl2 (43.69 g, 367.25 mmol, 26.64 mL) was stirred at 80 °C for 2 h. The mixture was then concentrated to remove SOCl2. MeOH (11 mL) was added. The mixture was stirred at 20 °C for 0.5 h. It was poured into water (600 mL) and extracted with EA (300 mL x 3). The combined organic layers were washed with brine (200 mL), dried over Na2SO4, filtered, and concentrated to give methyl 4-bromo-3-chlorosulfonylbenzoate (10 g, crude) as a yellow solid.

[0723] LCMS(ESI)m / z:[Br 79 M+H] + =314.8.

[0724] 1 H NMR (400MHz, DMSO-d6) δ = 9.31 (br s, 2H), 8.71-8.31 (m, 1H), 7.89-7.62 (m, 2H), 3.86 (s, 3H).

[0725] Step 3: Preparation of methyl 3-(allylaminosulfonyl)-4-bromobenzoate

[0726] DIEA (6.60 g, 51.03 mmol, 8.89 mL) was added to a solution of methyl 4-bromo-3-chlorosulfonylbenzoate (4 g, 12.76 mmol) and propen-2-en-1-amine (1.31 g, 14.03 mmol, 1.73 mL, HCl) in DCM (40 mL) at 0 °C. The mixture was then stirred at 25 °C for 2 h. It was poured into water (100 mL) and extracted with DCM (60 mL x 3). The combined organic layers were washed with brine (60 mL), dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel rapid chromatography (SLC). 80g Rapid column chromatography using silica gel, eluent gradient of 0–50% ethyl acetate / petroleum ether @ 100 mL / min. Vacuum concentration fractionation yielded methyl 3-(allylaminosulfonyl)-4-bromobenzoate (4.1 g, 12.27 mmol, 96% yield) as a white solid.

[0727] 1 H NMR (400MHz, DMSO-d6) δ = 8.51-8.42 (m, 1H), 8.29 (br s,1H),8.01(d,J=0.8Hz,2H),5.77-5.52(m,1H),5.17-5.06(m,1H),5.03-4.93(m,1H),3.89(s,3H),3.57(brd,J=4.8Hz,2H)ppm.

[0728] Step 4: Preparation of methyl 3-(allylaminosulfonyl)-4-vinylbenzoate

[0729] Methyl 3-(allylaminosulfonyl)-4-bromobenzoate (3.1 g, 9.28 mmol), potassium trifluoro(vinyl)borate (6.21 g, 46.38 mmol), di-tert-butyl(cyclopentyl)phosphine, palladium dichloroisocyanurate, iron (604.6 mg, 0.928 mmol), and K3PO4 (5.91 g, 27.8 mmol) in dioxane (30 mL) and H2O (6 mL) were degassed and purged three times with N2. The mixture was stirred at 60 °C under N2 atmosphere for 16 h. It was poured into water (100 mL) and extracted with EA (60 mL x 3). The combined organic layers were washed with brine (60 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel rapid chromatography (SLC). 40g Silica gel rapid column chromatography, eluent with a 0–50% ethyl acetate / petroleum ether gradient @ 80 mL / min). Vacuum concentration and fractionation yielded methyl 3-(allylaminosulfonyl)-4-vinylbenzoate (1.5 g, 5.33 mmol, 57% yield) as a white solid. LCMS (ESI) m / z: [M+H] + =282.1.

[0730] 1 H NMR (400MHz, CDCl3) δ = 8.62 (d, J = 1.6Hz, 1H), 8.29-8.11 (m, 1H), 7.69 (d, J = 8.0Hz, 1H), 7.62-7.47 (m, 1 H),5.92-5.77(m,1H),5.73-5.53(m,2H),5.22-4.97(m,2H),4.84-4.57(m,1H),3.69-3.41(m,2H)ppm.

[0731] Step 5: Preparation of methyl 3-[allyl(methyl)aminosulfonyl]-4-vinylbenzoate

[0732] MeI (201.81 mg, 1.42 mmol, 88.5 μL) was added to a solution of methyl 3-(allylaminosulfonyl)-4-vinylbenzoate (200 mg, 0.711 mmol) and K₂CO₃ (196.5 mg, 1.42 mmol) in DMF (2 mL). The mixture was stirred at 20 °C for 3 h. It was poured into water (60 mL) and extracted with EA (30 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel rapid chromatography (SLC). 12g Silica gel rapid column chromatography, eluent with a 0–50% ethyl acetate / petroleum ether gradient @ 50 mL / min. Vacuum concentration fractionation yielded methyl 3-[allyl(methyl)aminosulfonyl]-4-vinylbenzoate (190 mg, 0.643 mmol, 90% yield) as a yellow oil. LCMS (ESI) m / z: [M+H] + =296.0

[0733] 1H NMR (400MHz, CDCl3) δ = 8.56 (d, J = 1.6Hz, 1H), 8.26-8.09 (m, 1H), 7.73 (d, J = 8.4Hz, 1H), 7.67-7.53 (m, 1H), 5.88-5.77 (m,1H),5.76-5.64(m,1H),5.59-5.50(m,1H),5.27-5.16(m,2H),3.96(s,3H),3.75(d,J=6.4Hz,2H),2.75(s,3H)ppm.

[0734] Step 6: Preparation of methyl 2-methyl-1,1-dioxo-3H-1λ6,2-benzothiazoheptan-8-carboxylate

[0735] A mixture of methyl 3-[allyl(methyl)aminosulfonyl]-4-vinylbenzoate (190 mg, 0.643 mmol) and benzylene-[1,3-bis(2,4,6-trimethylphenyl)imidazolidin-2-ylidene]-dichloro-ruthenium; tricyclohexylphosphane (54.61 mg, 64.33 μmol) in 10 mL of DCM was degassed and purged with N2 3x. The mixture was stirred at 25 °C under N2 atmosphere for 2 h. The residue was purified by silica gel rapid chromatography. 12g Silica gel rapid column chromatography, eluent with a ~50% ethyl acetate / petroleum ether gradient @ 30 mL / min). Vacuum concentration and fractionation yielded methyl 2-methyl-1,1-dioxo-3H-1λ6,2-benzothiazoheptaenoic acid-8-carboxylate (130 mg, 0.486 mmol, 76% yield), a white solid. LCMS (ESI) m / z: [M+H] + =268.0.

[0736] 1 H NMR (400MHz, DMSO-d6) δ = 8.38 (d, J = 2.0Hz, 1H), 8.26-8.10 (m, 1H), 7.78 (d, J = 8.4Hz, 1H), 6.72 (br d,J=13.2z,1H),6.31-5.96(m,1H),4.45-4.17(m,2H),3.90(s,3H),2.55(s,3H)ppm.

[0737] Step 7: Preparation of methyl 2-methyl-1,1-dioxo-4,5-dihydro-3H-1λ6,2-benzothiazoheptan-8-carboxylate

[0738] A mixture of methyl 2-methyl-1,1-dioxo-3H-1λ6,2-benzothiazaheptaeno-8-carboxylate (130 mg, 0.486 mmol) and Pd / C (13 mg, 10% purity) in MeOH (4 mL) was degassed and purged three times with H₂. The mixture was then stirred at 20 °C under an H₂ atmosphere for 2 h. After filtration and concentration, methyl 2-methyl-1,1-dioxo-4,5-dihydro-3H-1λ6,2-benzothiazaheptaeno-8-carboxylate (110 mg, 0.408 mmol, 84% yield) was given as a white solid. LCMS (ESI) m / z: [M+H] + =270.0.

[0739] 1 H NMR (400MHz, CDCl3) δ = 8.56 (d, J = 2.0Hz, 1H), 8.17-8.03 (m, 1H), 7.38 (d, J = 7.6Hz, 1H), 3.95(s,3H),3.92-3.59(m,2H),3.45-3.23(m,2H),2.65(s,3H),1.91-1.80(m,3H)ppm.

[0740] Step 8: Preparation of 2-methyl-1,1-dioxo-4,5-dihydro-3H-1λ6,2-benzothiazoheptan-8-carboxylic acid (intermediate 13)

[0741] To a solution of methyl 2-methyl-1,1-dioxo-4,5-dihydro-3H-1λ6,2-benzothiazoheptaenoic acid-8-carboxylic acid (110 mg, 0.408 mmol) in THF (1 mL) and H₂O (1 mL), LiOH·H₂O (68.56 mg, 1.63 mmol) was added. The mixture was stirred at 25 °C for 2 h. The solution was adjusted to H = 5 with aqueous HCl (1 M) and extracted with EA (20 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na₂SO₄, filtered, and concentrated to give 2-methyl-1,1-dioxo-4,5-dihydro-3H-1λ6,2-benzothiazoheptaenoic acid-8-carboxylic acid (80 mg, 0.313 mmol, 77% yield) as a white solid.

[0742] LCMS(ESI)m / z:[M+H] + =519.2

[0743] 1H NMR (400MHz, DMSO-d6) δ=8.27(d,J=1.6Hz,1H),8.14-8.01(m,1H),7.59(d,J=8.0Hz,1H),3.75-3.55(m,2H),3.23(br s,3H),2.55(s,3H),1.83-1.71(m,2H)ppm.

[0744] Intermediate 14,4-(difluoromethyl)-3-(methanesulfonyl)benzoic acid

[0745]

[0746] Step 1: Preparation of methyl 3-bromo-4-(difluoromethyl)benzoate

[0747] DAST (596.87 mg, 3.70 mmol, 489.24 μL) was added to a solution of methyl 3-bromo-4-carboxybenzoate (300 mg, 1.23 mmol) in DCM (3 mL). The mixture was stirred at 25 °C for 1 hr. The reaction mixture was diluted with saturated NaHCO3 (20 mL) and extracted with DCM (20 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated to give the residue. The residue was purified by column chromatography (petroleum ether / ethyl acetate = 1 / 0-3 / 1). The eluent was concentrated to give methyl 3-bromo-4-(difluoromethyl)benzoate (190 mg, 716.84 μmol, 58% yield) as a yellow oil.

[0748] 1 H NMR (400MHz, CDCl3) δ = 8.28 (s, 1H), 8.09 (d, J = 8.0Hz, 1H), 7.75 (d, J = 8.0Hz, 1H), 7.06-6.79 (m, 1H), 3.96 (s, 3H) ppm.

[0749] Step 2: Preparation of 3-bromo-4-(difluoromethyl)benzoic acid

[0750] To a solution of methyl 3-bromo-4-(difluoromethyl)benzoate (90 mg, 339.56 μmol) in THF / MeOH / H₂O = 2 / 1 / 1 (1 mL), NaOH (27.16 mg, 679.11 μmol) was added. The mixture was stirred at 30 °C for 2 h. The reaction mixture was diluted with 1 N HCl (10 mL) and extracted with DCM (10 mL x 2). The combined organic layers were dried over anhydrous Na₂SO₄, filtered, and concentrated to give 3-bromo-4-(difluoromethyl)benzoic acid (70 mg, 278.86 μmol, 82% yield) as a yellow oil, which was used directly for the next step.

[0751] Step 3: Preparation of 4-(difluoromethyl)-3-methylthioalkylbenzoic acid

[0752] A mixture of 3-bromo-4-(difluoromethyl)benzoic acid (50 mg, 0.199 mmol), DABCO (44.68 mg, 0.398 mmol, 44 μL), and CuI (37.93 mg, 0.199 mmol) in DMSO (0.5 mL) was stirred at 145 °C for 12 h. 1 N HCl was added to the mixture to adjust the pH to 5. The mixture was filtered. The filtrate was concentrated to give a residue. The residue was purified by reverse-phase chromatography (0.1% FA). The eluent was concentrated to give 4-(difluoromethyl)-3-methylthioalkylbenzoic acid (30 mg, 0.137 mmol, 69% yield) as a yellow solid. LCMS (ESI) m / z: [M+H] + =218.9.

[0753] 1 H NMR (400MHz, CDCl3) δ = 8.10 (s, 1H), 8.02 (d, J = 8.4Hz, 1H), 7.75 (d, J = 8.0Hz, 1H), 7.15-6.87 (m, 1H), 2.58 (s, 3H) ppm.

[0754] Step 4: Preparation of 4-(difluoromethyl)-3-methylsulfonylbenzoic acid (intermediate 14)

[0755] A mixture of Oxone (169.03 mg, 274.95 μmol) in H₂O (0.5 mL) was added to a solution of 4-(difluoromethyl)-3-methylthioalkylbenzoic acid (30 mg, 137.48 μmol) in MeOH (0.5 mL) at 0 °C. The mixture was stirred at 25 °C for 12 h. The reaction mixture was diluted with H₂O (10 mL) and extracted with DCM (10 mL). The organic layer was dried over anhydrous Na₂SO₄, filtered, and concentrated to give the residue. The residue was purified by reverse-phase (0.1% FA). The eluent was concentrated to give 4-(difluoromethyl)-3-(methylsulfonyl)benzoic acid (20 mg, 79.9 μmol, 58% yield) as a white solid.

[0756] LCMS(ESI)m / z:[M+H] + =250.9.

[0757] Intermediate 15,6-methyl-5-(methanesulfonyl)nicotinic acid

[0758]

[0759] Step 1: Preparation of 6-methyl-5-(meththio)nicotinic acid

[0760] Sodium thiomethoxide (320.30 mg, 1.95 mmol) was added to a solution of methyl 5-fluoro-6-methylpyridine-3-carboxylate (300 mg, 1.77 mmol) in DMF (2 mL). The mixture was stirred at 100 °C for 16 h. The reaction mixture was quenched with HCl (1 M) (40 mL) and extracted with EA / MeOH = 15 / 1 (40 mL x 5). The combined organic layers were dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by reverse-phase HPLC (0.1% FA conditions). The solution was concentrated under reduced pressure to remove MeCN and then lyophilized to give 6-methyl-5-(methylthio)nicotinic acid (200 mg, 1.09 mmol, 62% yield) as a yellow solid.

[0761] LCMS(ESI)m / z:[M+H] + =183.9.

[0762] 1 H NMR (400MHz, DMSO-d6) δ = 14.14-12.40 (m, 1H), 8.69 (d, J = 1.6Hz, 1H), 7.94 (d, J = 2.0Hz, 1H), 2.55 (s, 3H), 2.50 (s, 3H) ppm.

[0763] Step 2: Preparation of 6-methyl-5-(methanesulfonyl)nicotinic acid (intermediate 15)

[0764] Add to a solution of 6-methyl-5-(meththio)nicotinic acid (30 mg, 163.73 μmol) in MeOH (1 mL) (150.98 mg, 0.246 mmol) and H₂O (1 mL). The mixture was stirred at 25 °C for 16 h. The reaction mixture was dissolved in DMSO (5 mL), then filtered to obtain the filtrate. The filtrate was purified by reversed-phase HPLC (0.1% FA conditions). The solution was concentrated under reduced pressure to remove MeCN, and then lyophilized to give 6-methyl-5-(methanesulfonyl)nicotinic acid (15 mg, 67.8 μmol, 41% yield) as a white solid. LCMS (ESI) m / z: [M+H] + =216.1.

[0765] 1H NMR (400MHz, DMSO-d6) δ = 15.43-11.63 (m, 1H), 9.17 (d, J = 2.0Hz, 1H), 8.61 (d, J = 2.0Hz, 1H), 3.36 (s, 3H), 2.90 (s, 3H) ppm.

[0766] Intermediate 16,3-chloro-4-methyl-5-methylsulfonyl-benzoic acid

[0767]

[0768] Step 1: Preparation of 3-chloro-5-chlorosulfonyl-4-methylbenzoic acid

[0769] A mixture of 3-chloro-4-methylbenzoic acid (1 g, 5.86 mmol) and chlorosulfonic acid (10.25 g, 87.93 mmol, 5.85 mL) was stirred at 120 °C for 12 h. The reaction mixture was then added to H₂O (20 mL) at 0 °C. A white solid precipitated from the mixture. The solid was collected by filtration and dried under reduced pressure to give 3-chloro-5-chlorosulfonyl-4-methylbenzoic acid (1.2 g, 4.46 mmol, 76% yield) as a white solid.

[0770] 1 H NMR (400MHz, DMSO-d6) δ = 8.30 (d, J = 2.0Hz, 1H), 7.85 (d, J = 2.0Hz, 1H), 2.63 (s, 3H) ppm.

[0771] Step 2: Preparation of 3-chloro-4-methyl-5-methylsulfonyl-benzoic acid (intermediate 16)

[0772] 3-Chloro-5-chlorosulfonyl-4-methylbenzoic acid (300 mg, 1.11 mmol) was added to a solution of Na₂SO₃ (140.51 mg, 1.11 mmol) and NaHCO₃ (280.97 mg, 3.34 mmol, 130.08 μL) in H₂O (1.2 mL) at 80 °C. The mixture was stirred at 80 °C for 1 hr. Then, 2-bromoacetic acid (309.8 mg, 2.23 mmol, 161 μL) and NaOH (89.19 mg, 2.23 mmol) were added, and the mixture was stirred at 110 °C for 12 h. The reaction mixture was diluted with H₂O (10 mL), and then 1 N HCl was added to adjust the pH to 3. A white solid precipitated from the mixture. The solid was collected by filtration and dried under reduced pressure to obtain 3-chloro-4-methyl-5-methylsulfonylbenzoic acid (120 mg, 0.483 mmol, 43% yield), a white solid. LCMS (ESI) m / z: [M+H] +=248.9

[0773] 1 H NMR (400MHz, DMSO-d6) δ = 8.41 (d, J = 1.6Hz, 1H), 8.21 (d, J = 1.6Hz, 1H), 3.32 (s, 3H), 2.74 (s, 3H) ppm.

[0774] Intermediate 17,4-chloro-3-fluoro-5-methylsulfonyl-benzoic acid

[0775]

[0776] Step 1: Preparation of 4-chloro-3-fluoro-5-methylthioalkylbenzoic acid

[0777] A mixture of methyl 3-bromo-4-chloro-5-fluorobenzoate (200 mg, 747.72 μmol), CuI (142.40 mg, 747.72 μmol), and DABCO (167.8 mg, 1.50 mmol, 164 μL) in DMSO (2 mL) was stirred at 145 °C for 12 h under a N2 atmosphere. The reaction mixture was filtered. The filtrate was purified by reversed-phase HPLC (0.1% FA conditions). The desired fraction was lyophilized to give 4-chloro-3-fluoro-5-methylthioalkylbenzoic acid (90 mg, 0.371 mmol, 50% yield) as a white solid. LCMS (ESI) m / z: [M+H] + =220.9. 1 H NMR (400MHz, DMSO-d6) δ = 7.66-7.56 (m, 2H), 2.59 (s, 3H) ppm.

[0778] Step 2: Preparation of 4-chloro-3-fluoro-5-methylsulfonyl-benzoic acid (intermediate 17)

[0779] Add to a solution of 4-chloro-3-fluoro-5-methylthioalkylbenzoic acid (90 mg, 0.408 mmol) in H₂O (1 mL) and MeOH (2 mL) (501.5 mg, 0.816 mmol). The reaction system was stirred at 20 °C under a N2 atmosphere for 12 h. A saturated aqueous solution of Na2SO3 (5 mL) was added to the mixture. The mixture was extracted with EA (5 mL x 3). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, filtered, and concentrated to give 4-chloro-3-fluoro-5-methylsulfonylbenzoic acid (40 mg, 0.158 mmol, 39% yield) as a white solid. LCMS (ESI) m / z: [M+H] + =252.9.

[0780] 1 H NMR (400MHz, DMSO-d6) δ = 8.35 (s, 1H), 8.21-8.19 (m, 1H), 3.45 (s, 3H) ppm.

[0781] Intermediate 18. ((2-chloro-1,6-naphthid-7-yl)methyl)tert-butyl carbamate

[0782]

[0783] Step 1. Preparation of 2-bromo-5-iodo-pyridine-4-amine

[0784] NIS (93.6 g, 416 mmol) was added to a solution of 2-bromopyridine-4-amine (60 g, 347 mmol) in MeCN (1.5 L) at 80 °C. The reaction mixture was stirred at 80 °C for 36 h. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was diluted with saturated Na₂SO₃ (1.5 L) and extracted with EA (1.5 L x 2). The combined organic layers were washed with brine (1 L), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO₂, PE / EA = 20:3) and concentrated under reduced pressure to give 2-bromo-5-iodopyridine-4-amine (65 g, 217 mmol) as a pale yellow solid.

[0785] 1 H NMR (400MHz, CDCl3) δ = 8.31 (s, 1H), 6.79 (s, 1H), 4.75 (br s, 2H) ppm.

[0786] Step 2.3 Preparation of ethyl 4-(4-amino-6-bromo-3-pyridyl)prop-2-enoate

[0787] Ethyl propionic acid (45.1 mL, 415 mmol), Et3N (43.3 mL, 311 mmol), Pd(OAc)2 (2.3 g, 10.4 mmol), and tris-o-tolylphosphane (6.3 g, 20.7 mmol) were added to a solution of 2-bromo-5-iodopyridin-4-amine (62 g, 207 mmol) in DMF (620 mL). The mixture was stirred at 100 °C for 3 h. The reaction mixture was diluted with water (4 L) and extracted with EA (2 L x 2). The combined organic layers were washed with brine (2 L), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, PE / EA = 20:3), and concentrated under reduced pressure to obtain ethyl 3-(4-amino-6-bromo-3-pyridyl)prop-2-enoate (50 g, 170 mmol), which was a pale yellow solid.

[0788] LCMS(ESI)m / z:[79BrM+H]+=271.1.

[0789] 1 H NMR (400MHz, DMSO-d6) δ = 8.23 ​​(s, 1H), 7.73 (d, J = 16.0Hz, 1H), 6.90-6.67 (m, 3H), 6.52 (d, J = 16.0Hz, 1H), 4.18 (d, J = 7.2Hz, 2H), 1.25 (d, J = 7.2Hz, 3H) ppm.

[0790] Step 3.7 Preparation of bromo-1,6-naphthidium-2(1H)-one

[0791] Sodium thiomethoxide (24.2 mL, 380 mmol) was added to a solution of ethyl 3-(4-amino-6-bromo-3-pyridyl)prop-2-enoate (40 g, 148 mmol) in EtOH (200 mL). The reaction mixture was stirred at 60 °C for 2 h. The reaction mixture was diluted with water (400 mL) and then neutralized with 1 N HCl to pH 7.0. The solid was filtered off, and the filter cake was washed with water (50 mL). The filter cake was concentrated under reduced pressure to give 7-bromo-1,6-naphthidium-2(1H)-one (22 g, 96.8 mmol) as a yellowish-white solid.

[0792] LCMS(ESI)m / z:[79BrM+H]+=224.9.

[0793] 1H NMR (400MHz, DMSO-d6) δ = 12.08 (br s, 1H), 8.65 (s, 1H), 7.99 (d, J = 9.6Hz, 1H), 7.36 (s, 1H), 6.62 (d, J = 9.6Hz, 1H) ppm.

[0794] Step 4.2 Preparation of oxo-1,2-dihydro-1,6-naphthyl-7-formonitrile

[0795] Zinc powder (406.80 mg, 6.22 mmol) was added to 7-bromo-1,6-naphthidium-2(1H)-one (7 g, 31.1 mmol), Zn(CN)₂ (3.95 mL, 62.2 mmol), and Pd(dppf)Cl₂. . CH₂Cl₂ (5.08 g, 6.22 mmol) was dissolved in DMA (140 mL). The reaction mixture was degassed, purged three times with N₂, and then stirred at 120 °C for 2 h. The reaction mixture was diluted with water (200 mL) and extracted with DCM / isopropanol (v / v = 3:1) (150 mL x 2). The combined organic layers were filtered, washed with brine (200 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO₂, PE / EA = 1:1), and concentrated under reduced pressure to give 2-oxo-1,2-dihydro-1,6-naphthyl-7-carboxynitrile (3 g, 17.5 mmol) as a yellowish-white solid.

[0796] 1 H NMR (400MHz, DMSO-d6) δ = 12.37 (br s, 1H), 8.97 (s, 1H), 8.09 (d, J = 9.6Hz, 1H), 7.67 (s, 1H), 6.77 (d, J = 9.6Hz, 1H) ppm.

[0797] Step 5. Preparation of 2-chloro-1,6-naphthyl-7-carboxynitrile

[0798] A mixture of 2-oxo-1,2-dihydro-1,6-naphthyl-7-carboxynitrile (3.0 g, 17.5 mmol) and POCl3 (30 mL, 323 mmol) was stirred at 80 °C for 2 h. The reaction mixture was poured into H2O (2 L) and the pH was adjusted to 7 with NaHCO3. The solution was extracted with EA (1.5 L x 2), the combined organic layers were washed with brine (2 L), dried over Na2SO4, filtered, and concentrated under reduced pressure to give 2-chloro-1,6-naphthyl-7-carboxynitrile (1.1 g, 5.76 mmol) as a brown solid. LCMS (ESI) m / z: [M+H]+ = 190.1.

[0799] 1 H NMR (400MHz, DMSO-d6) δ = 9.58 (d, J = 0.8Hz, 1H), 8.79 (d, J = 0.8Hz, 1H), 8.69 (s, 1H), 8.00 (d, J = 8.8Hz, 1H) ppm.

[0800] Step 6. Preparation of (2-chloro-1,6-naphthid-7-yl)methylamine

[0801] DIBAL-H (1M, 329.64mL, 2.5eq) was added dropwise to a solution of 2-chloro-1,6-naphthid-7-carboxynitrile (25g, 131.86mmol) in DCM (1000mL) under a N2 atmosphere at -70°C. The reaction mixture was stirred at -70°C for 2h. The reaction mixture was quenched with water (500mL) and saturated sodium potassium tartrate (1500mL), and stirred for another 30min. The mixture was extracted with DCM:MeOH = 10:1 (6000mL x 3). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give (2-chloro-1,6-naphthid-7-yl)methylamine (51g, crude), a brown solid, which was used directly for the next step. LCMS(ESI) m / z: [ 35 ClM+H] + =194.2.

[0802] Step 7: Preparation of ((2-chloro-1,6-naphthid-7-yl)methyl)tert-butyl carbamate (intermediate 18)

[0803] To a solution of (2-chloro-1,6-naphthid-7-yl)methylamine (51 g, 263.4 mmol) in DCM (1500 mL), (Boc)₂O (172.45 g, 790.16 mmol) and DIEA (102.12 g, 790.16 mmol) were added. The mixture was stirred at 25 °C for 16 h. The reaction mixture was diluted with water (1500 mL) and then filtered. The filtrate was extracted with DCM (1000 mL x 3). The combined organic layers were washed with brine (1500 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue (SiO2, petroleum ether / ethyl acetate = 10 / 1-2 / 1-1 / 3) was purified by column chromatography, concentrated and eluted under reduced pressure to give tert-butyl ((2-chloro-1,6-naphthid-7-yl)methyl)carbamate (21 g, 64.34 mmol, 24% yield) as a pale yellow solid.

[0804] LCMS(ESI)m / z:[M+H]+=293.9.

[0805] 1H NMR (400MHz, DMSO-d6) δ = 9.37 (s, 1H), 8.62 (d, J = 8.4Hz, 1H), 7.71 (d, J = 8.4Hz, 1 H),7.58-7.53(m,2H),4.39(d,J=6.4Hz,2H),4.20–4.25(m,2H),1.41(s,9H)ppm.

[0806] Intermediate 19: [2-[6-(2,2-difluorocyclopropyl)-2-pyridyl]-1,6-naphthid-7-yl]methylamine

[0807]

[0808] Step 1: Preparation of 2-bromo-6-(2,2-difluorocyclopropyl)pyridine

[0809] A solution of TMSCF3 (1.55 g, 10.87 mmol) in THF (1 mL) was added to a mixture of 2-bromo-6-vinylpyridine (500 mg, 2.72 mmol) and NaI (81.45 mg, 0.543 mmol) in THF (4 mL) at 70 °C under a N2 atmosphere for 1 h. The mixture was then stirred at 70 °C under a N2 atmosphere for 1 h. The residue was purified by silica gel chromatography (PE–PE / EA = 50 / 1). The eluent was concentrated under reduced pressure to give 2-bromo-6-(2,2-difluorocyclopropyl)pyridine (570 mg, 2.44 mmol, 90% yield) as a yellow oil. LCMS (ESI) m / z: [M+H] + =233.9.

[0810] 1 H NMR (400MHz, CDCl3) δ=7.52-7.48(m,1H),7.39-7.37(m,1H),7.19(d,J=7.6Hz,1H),2.95-2.84(m,1H),2.21-2.12(m,1H),1.89-1.83(m,1H)ppm.

[0811] Step 2: Preparation of [6-(2,2-difluorocyclopropyl)-2-pyridyl]-trimethylstannane

[0812] A mixture of 2-bromo-6-(2,2-difluorocyclopropyl)pyridine (100 mg, 427.28 μmol), hexamethyldistin (279.97 mg, 854.55 μmol, 177.20 μL), and Pd(PPh3)4 (49.37 mg, 42.73 μmol) in dioxane (2 mL) was stirred at 100 °C under a N2 atmosphere for 2 h. The mixture was filtered and concentrated under reduced pressure to give [6-(2,2-difluorocyclopropyl)-2-pyridyl]-trimethylstanane (170 mg, crude), as a brown oil.

[0813] LCMS(ESI)m / z:[M+H] + =320.1.

[0814] Step 3: Preparation of N-[[2-[6-(2,2-difluorocyclopropyl)-2-pyridyl]-1,6-naphthid-7-yl]methyl]tert-butyl carbamate

[0815] A mixture of ((2-chloro-1,6-naphthid-7-yl)methyl)carbamate tert-butyl ester (50 mg, 170.21 μmol), [6-(2,2-difluorocyclopropyl)-2-pyridyl]trimethylstanane (163 mg, 0.511 mmol), and Pd(PPh3)2Cl2 (11.95 mg, 17.02 μmol) in dioxane (1 mL) was stirred at 100 °C under a nitrogen atmosphere for 16 h. The mixture was then poured into saturated KF (10 mL) and stirred at 20 °C for 30 min. The mixture was extracted with EA (10 mL x 3). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel chromatography (PE / EA = 10 / 1 - EA). The eluent was concentrated under reduced pressure to give N-[[2-[6-(2,2-difluorocyclopropyl)-2-pyridyl]-1,6-naphthid-7-yl]methyl]tert-butyl carbamate (30 mg, 72.74 μmol, 43% yield), as a yellow solid. LCMS (ESI) m / z: [M+H] + =413.3.

[0816] Step 4: Preparation of [2-[6-(2,2-difluorocyclopropyl)-2-pyridyl]-1,6-naphthid-7-yl]methylamine (intermediate 19)

[0817] TFA (462 mg, 4.05 mmol, 0.3 mL) was added to a mixture of N-[[2-[6-(2,2-difluorocyclopropyl)-2-pyridyl]-1,6-naphthid-7-yl]methyl]carbamate (30 mg, 72.74 μmol) in DCM (1 mL) at 0 °C. The mixture was stirred at 30 °C for 1 hr. The mixture was concentrated under reduced pressure to give [2-[6-(2,2-difluorocyclopropyl)-2-pyridyl]-1,6-naphthid-7-yl]methylamine (31 mg, 72.71 μmol, 100% yield, TFA salt) as a yellow solid. LCMS (ESI) m / z: [M+H] + =313.2.

[0818] Intermediate 20. (2-(2-(2,2-difluorocyclopropyl)pyrimidin-4-yl)-1,6-naphthidin-7-yl)methanamine ester

[0819]

[0820] Step 1: Preparation of 2,2-difluorocyclopropanemimide (carboximidamide)

[0821] A solution of Al(CH3)3 (2M, 64.29 mL) was added to a mixture of NH4Cl (6.88 g, 128.59 mmol) in toluene (50 mL) at 0 °C. The mixture was then stirred at 25 °C for 1 hr. Methyl 2,2-difluorocyclopropanecarboxylate (3.5 g, 25.72 mmol) was added to the solution at 0 °C, and the solution was stirred at 80 °C for 12 h. A large amount of white precipitate formed. The reaction mixture was cooled to 0 °C. MeOH (50 mL) was added, and the mixture was stirred for 10 min. The mixture was filtered. The filtrate was concentrated under vacuum to give 2,2-difluorocyclopropanemethylimine amide (3 g, crude), a white solid, which was used directly.

[0822] Step 2: Preparation of 2-(2,2-difluorocyclopropyl)pyrimidin-4-ol

[0823] K₂CO₃ (6.90 g, 49.94 mmol) was added in a single batch to a mixture of 2,2-difluorocyclopropane-methylimine amide (3.00 g, 24.97 mmol) in EtOH (40 mL) under a nitrogen atmosphere at 25 °C. The mixture was stirred at 25 °C for 10 min, and then (E)-3-ethoxyethyl acrylate (1.2 g, 8.32 mmol, 1.20 mL) was added at 25 °C. The mixture was stirred at 75 °C for 6 h. The reaction mixture was filtered, and the filtrate was concentrated under vacuum. The mixture was purified by silica gel chromatography (DCM / MeOH = 20 / 1). The eluent was concentrated to give 2-(2,2-difluorocyclopropyl)pyrimidin-4-ol (500 mg, 2.90 mmol, 35% yield) as a white solid.

[0824] LCMS(ESI)m / z:[M+H] + =173.2.

[0825] 1 H NMR (400MHz, CDCl3) δ=8.04-7.95(m,1H),6.43-6.35(m,1H),2.84-2.69(m,1H),2.51-2.39(m,1H),2.00-1.88(m,1H)ppm.

[0826] Step 3: Preparation of 4-chloro-2-(2,2-difluorocyclopropyl)pyrimidine

[0827] Oxaloyl chloride (516 mg, 4.07 mmol, 356 μL) was added in a single batch to a mixture of 2-(2,2-difluorocyclopropyl)pyrimidin-4-ol (350 mg, 2.03 mmol) and DMF (14.9 mg, 0.203 mmol, 15.6 μL) in DCM (6 mL) under a nitrogen atmosphere at 0 °C. The mixture was stirred at 25 °C for 20 min. The mixture was then added to saturated NaHCO3 (50 mL). The aqueous phase was extracted with DCM (50 mL x 2). The combined organic phases were washed with brine (50 mL x 1), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (PE / EA = 10 / 1). The eluent was concentrated to give 4-chloro-2-(2,2-difluorocyclopropyl)pyrimidin (150 mg, 0.787 mmol, 39% yield) as a pale yellow oil.

[0828] LCMS(ESI)m / z:[M+H] + =190.9,192.9.

[0829] Step 4: Preparation of 2-(2,2-difluorocyclopropyl)-4-(tributylmethyltinyl)pyrimidine

[0830] Pd(PPh3)4 (60.63 mg, 52.47 μmol) was added in a single batch to a mixture of 4-chloro-2-(2,2-difluorocyclopropyl)pyrimidine (100 mg, 0.525 mmol) and trimethyl(trimethylstanyl)stanane (343.8 mg, 1.05 mmol, 218 μL) in dioxane (2 mL) under a nitrogen atmosphere at 25 °C. The mixture was stirred at 100 °C for 2 h. The mixture was then poured into water (10 mL) and extracted with ethyl acetate (10 mL x 2). The combined organic phases were washed with brine (10 mL x 1), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum to give 2-(2,2-difluorocyclopropyl)-4-(tributylstanyl)pyrimidine (150 mg, crude) as a yellow oil.

[0831] LCMS(ESI)m / z:[M+H] + =320.9.

[0832] Step 5: Preparation of tert-butyl ((2-(2-(2,2-difluorocyclopropyl)pyrimidin-4-yl)-1,6-naphthidin-7-yl)methyl)carbamate

[0833] Pd(PPh3)2Cl2 (21.51 mg, 30.64 μmol) was added in a single batch to a mixture of 2-(2,2-difluorocyclopropyl)-4-(tributylmethylenetinyl)pyrimidine (147 mg, 0.460 mmol) and ((2-chloro-1,6-naphthid-7-yl)methyl)carbamate (90 mg, 0.306 mmol) in dioxane (2 mL) under a nitrogen atmosphere at 25 °C. The mixture was stirred at 100 °C for 12 h. The mixture was then poured into water (30 mL) and extracted with ethyl acetate (20 mL x 2). The combined organic phases were washed with brine (20 mL x 1), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (PE / EA = 3 / 1). The eluent was concentrated to give tert-butyl ((2-(2-(2,2-difluorocyclopropyl)pyrimidin-4-yl)-1,6-naphthidin-7-yl)methyl)carbamate (90 mg, 0.218 mmol, 71% yield), as a yellow solid. LCMS (ESI) m / z: [M+H] + =414.0.

[0834] 1H NMR (400MHz, DMSO-d6)δ=9.50-9.43(m,1H),9.08-9.01(m,1H),8.84-8.78(m,1H),8.73-8.67(m,1H),8.50-8.44(m,1H) ,7.86-7.80(m,1H),7.72-7.60(m,1H),4.51-4.42(m,2H),2.30-2.13(m,1H),1.52-1.41(m,9H),1.41-1.21(m,2H)ppm.

[0835] Step 6: Preparation of (2-(2-(2,2-difluorocyclopropyl)pyrimidin-4-yl)-1,6-naphthidin-7-yl)carbamate

[0836] TFA (770.0 mg, 6.75 mmol, 500 μL) was added in a single batch to a mixture of ((2-(2-(2,2-difluorocyclopropyl)pyrimidin-4-yl)-1,6-naphthidin-7-yl)methyl)carbamate (90 mg, 0.218 mmol) in DCM (1 mL) at 25 °C under a nitrogen atmosphere. The mixture was stirred at 25 °C for 30 min. The mixture was then poured into ice-water (20 mL) and extracted with ethyl acetate (20 mL x 1). The organic phase was discarded. Saturated NaHCO3 was added to the aqueous phase to adjust the pH to 8. The aqueous phase was then extracted with ethyl acetate (20 mL x 2). The combined organic phases were washed with brine (10 mL*1), dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to obtain (2-(2-(2,2-difluorocyclopropyl)pyrimidin-4-yl)-1,6-naphthidin-7-yl)carbamate (70 mg, crude), which was a pale yellow solid and used directly without purification.

[0837] Intermediate 21. [2-[6-(2,2-difluoro-1-methyl-cyclopropyl)-2-pyridyl]-1,6-naphthid-7-yl]methylamine

[0838] Step 1: Preparation of 2-bromo-6-(2,2-difluoro-1-methyl-cyclopropyl)pyridine

[0839] TMSCF3 (287.19 mg, 2.02 mmol) was added dropwise to a mixture of 2-bromo-6-isopropenylpyridine (100 mg, 504.90 μmol) and NaI (15.14 mg, 100.98 μmol) in THF (0.8 mL) for 30 min at 70 °C under a nitrogen atmosphere. The mixture was then stirred at 70 °C under a nitrogen atmosphere for 30 min. The mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel chromatography (PE–PE / EA = 20 / 1). The eluent was concentrated under reduced pressure to give 2-bromo-6-(2,2-difluoro-1-methyl-cyclopropyl)pyridine (125 mg, 0.504 mmol, 100% yield) as a yellow oil. LCMS (ESI) m / z: [M+H] + =247.9.

[0840] 1 H NMR (400MHz, CDCl3) δ=7.56-7.50(m,1H),7.40-7.37(m,1H),7.30(d,J=7.6Hz,1H),2.28-2.21(m,1H),1.63-1.59(m,3H),1.48-1.41(m,1H)ppm.

[0841] Step 2: Preparation of [6-(2,2-difluoro-1-methylcyclopropyl)-2-pyridyl]-trimethyl-stanane

[0842] A mixture of 2-bromo-6-(2,2-difluoro-1-methylcyclopropyl)pyridine (100 mg, 403.12 μmol), hexamethyldistin (264.15 mg, 0.806 mmol, 167 μL), and Pd(PPh3)4 (46.58 mg, 40.31 μmol) in dioxane (2 mL) was stirred at 100 °C under a nitrogen atmosphere for 2 h. The mixture was filtered, and the filtrate was concentrated under reduced pressure to give [6-(2,2-difluoro-1-methylcyclopropyl)-2-pyridyl]-trimethyl-stanane (210 mg, crude), a dark brown oil. LCMS (ESI) m / z: [M+H] + =334.0.

[0843] Step 3: Preparation of N-[[2-[6-(2,2-difluoro-1-methyl-cyclopropyl)-2-pyridyl]-1,6-naphthid-7-yl]methyl]tert-butyl carbamate

[0844] A mixture of ((2-chloro-1,6-naphthidin-7-yl)methyl)carbamate tert-butyl ester (60 mg, 0.204 mmol), [6-(2,2-difluoro-1-methylcyclopropyl)-2-pyridyl]trimethyl-stanane (203.4 mg, 0.613 mmol), and Pd(PPh3)2Cl2 (14.34 mg, 20.43 μmol) in dioxane (1 mL) was stirred at 100 °C under a nitrogen atmosphere for 16 h. The mixture was then poured into saturated KF (10 mL) and stirred at 20 °C for 30 min. The mixture was extracted with EA (10 mL x 3). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel chromatography (PE / EA = 10 / 1 - EA). The eluent was concentrated under reduced pressure to give N-[[2-[6-(2,2-difluoro-1-methyl-cyclopropyl)-2-pyridyl]-1,6-naphthid-7-yl]methyl] tert-butyl carbamate (42 mg, 98.49 μmol, 48% yield), as a yellow solid.

[0845] LCMS(ESI)m / z:[M+H] + =427.0.

[0846] Step 4: Preparation of [2-[6-(2,2-difluoro-1-methyl-cyclopropyl)-2-pyridyl]-1,6-naphthid-7-yl]methylamine (intermediate 21)

[0847] TFA (462.0 mg, 4.05 mmol, 0.3 mL) was added to a solution of N-[[2-[6-(2,2-difluoro-1-methyl-cyclopropyl)-2-pyridyl]-1,6-naphthid-7-yl]methyl]carbamate (42 mg, 98.49 μmol) in DCM (1 mL) at 0 °C. The mixture was stirred at 25 °C for 1 hr. The mixture was concentrated under reduced pressure to give [2-[6-(2,2-difluoro-1-methyl-cyclopropyl)-2-pyridyl]-1,6-naphthid-7-yl]methylamine (43 mg, 97.65 μmol, 99% yield, TFA salt) as a yellow solid. LCMS (ESI) m / z: [M+H] + =327.0.

[0848] Intermediate 22,1-imino-1-oxo-3,5-dihydro-2H-4,1λ6-benzoxazothioneine-8-carboxylic acid

[0849] Step 1: Preparation of N-(8-bromo-1-oxo-3,5-dihydro-2H-4,1λ6-benzoxazothionein-1-ylidene)-2,2,2-trifluoroacetamide

[0850] A mixture of 8-bromo-3,5-dihydro-2H-4,1λ4-benzoxazothiazine 1-oxide (50 mg, 191.47 μmol), 2,2,2-trifluoroacetamide (64.93 mg, 574.42 μmol), [acetoxy(phenyl)-λ3-iodanyl]acetate (129.51 mg, 402.09 μmol), and MgO (46.30 mg, 1.15 mmol) in DCM (3 mL) was stirred at 25 °C for 5 min. Then, rhodium diacetoxy (8.46 mg, 19.15 μmol) was added to the mixture, and the mixture was stirred at 25 °C under a N2 atmosphere for 16 h. The reaction mixture was diluted with MeOH (3 mL) to give N-(8-bromo-1-oxo-3,5-dihydro-2H-4,1λ6-benzoxazothionein-1-ylidene)-2,2,2-trifluoroacetamide (71 mg, 190.78 μmol, 100% yield), a yellow liquid, which was used directly in the next step. LCMS (ESI) m / z = [M + H +＝ 373.2.

[0851] Step 2: Preparation of 8-bromo-1-imino-3,5-dihydro-2H-4,1λ6-benzoxazine 1-oxide

[0852] K₂CO₃ (181.97 mg, 1.32 mmol) was added to a mixture of N-(8-bromo-1-oxo-3,5-dihydro-2H-4,1λ6-benzoxazothionein-1-ylidene)-2,2,2-trifluoroacetamide (70 mg, 188.09 μmol) in MeOH (3 mL), and the mixture was stirred at 25 °C for 4 h. The mixture was diluted with water (10 mL) and filtered to remove the precipitate. The filtrate was separated, and the aqueous layer was extracted with DCM (10 mL). The combined organic phases were washed with brine (10 mL), dried over anhydrous Na₂SO₄, filtered, and the filtrate was concentrated under vacuum. The residue (SiO2, PE:EtOAc = 20:1-1:1) was purified by column chromatography to give 8-bromo-1-imino-3,5-dihydro-2H-4,1λ6-benzoxazothiazine 1-oxide (40 mg, 137.65 μmol, 73% yield) as a white solid.

[0853] LCMS(ESI)m / z=[M+H] + =277.2.

[0854] 1H NMR (400MHz, DMSO_d6) δ = 8.07 (d, J = 2.0Hz, 1H), 7.81-7.79 (m, 1H), 7.46 (d, J = 8.0Hz, 1H), 4.99-4.81 (m, 3H), 4.21-4.13 (m, 2H), 3.42-3.39 (m, 2H) ppm.

[0855] Step 3: Preparation of 1-imino-1-oxo-3,5-dihydro-2H-4,1λ6-benzoxazheptaphylline-8-carboxylic acid

[0856] To a mixture of 8-bromo-1-imino-3,5-dihydro-2H-4,1λ6-benzoxazothiazine 1-oxide (40 mg, 144.85 μmol) and diacetoxypalladium (3.25 mg, 14.48 μmol) in DMSO (3 mL) and H₂O (0.3 mL), K₂CO₃ (30.03 mg, 217.27 μmol) and dicyclohexyl(3-dicyclohexylphosphoniumpropyl)phosphonium; bis(tetrafluoroborate) (17.74 mg, 28.97 μmol) were added. The mixture was degassed, purged three times with CO, and then stirred at 100 °C under a CO atmosphere (15 psi) for 4 h. The mixture was poured into water (50 mL), extracted with EA (20.0 mL x 2), and the combined organic layers were discarded. The aqueous layer was adjusted to pH 5 with HCl (1M), and then extracted with DCM (20.0 mL * 3). The combined organic phases were washed with brine (50.0 mL * 2), dried over Na2SO4, filtered, and the filtrate was evaporated to dryness to give 1-imino-1-oxo-3,5-dihydro-2H-4,1λ6-benzoxazothioneine-8-carboxylic acid (34 mg, crude), as a yellow solid.

[0857] Example 2. N-[[2-[6-(azacyclobutan-1-yl)-2-pyridyl]-1,6-naphthidin-7-yl]methyl]-1,1-dioxo-3,5-dihydro-2H-4,1λ6-benzoxazothionein-8-carboxamide]

[0858]

[0859] Step 1. Preparation of [6-(azacyclobutane-1-yl)-2-pyridyl]-trimethyl-stanane

[0860] To a solution of 2-(azacyclobutan-1-yl)-6-bromopyridine (150 mg, 703.98 μmol) in dioxane (3 mL), hexamethyldistin (461.28 mg, 1.41 mmol) and Pd(PPh3)4 (81.35 mg, 70.40 μmol) were added. The mixture was purged with N2 3x and then stirred at 100 °C under N2 atmosphere for 2 h. The reaction mixture was diluted with H2O (200 mL) and extracted with EA (150 mL x 3). The combined organic layers were washed with brine (200 mL), dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to give [6-(azacyclobutan-1-yl)-2-pyridyl]-trimethyl-stanane (209 mg, crude), a brown oil, which was used for the next step without further purification. LCMS (ESI) m / z: [M+H] + =299.3.

[0861] Step 2. Preparation of N-[[2-[6-(azacyclobutan-1-yl)-2-pyridyl]-1,6-naphthid-7-yl]methyl]tert-butyl carbamate

[0862] To a solution of N-[(2-chloro-1,6-naphthidin-7-yl)methyl]carbamate tert-butyl (100 mg, 340.43 μmol) in dioxane (2 mL), add [6-(azacyclobutan-1-yl)-2-pyridinyl]trimethyl-stanane (202.2 mg, 680.7 μmol) and Pd(PPh3)2Cl2 (23.9 mg, 34.04 μmol). The mixture was purged three times with N2 and then stirred at 100 °C under N2 atmosphere for 12 h. The reaction mixture was diluted with H2O (20 mL) and extracted with EA (30 mL x 3). The combined organic layers were washed with brine (30 mL), dried over Na₂SO₄, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 10:1–1:1) to give N-((2-(6-(azacyclobutan-1-yl)pyridin-2-yl)-1,6-naphthidin-7-yl)methyl)tert-butyl carbamate (70 mg, 173.45 μmol, 51% yield) as a yellow solid. LCMS (ESI) m / z: [M+H] + =392.4; 1H NMR (400MHz, CDCl3) δ=9.22(s,1H),8.68(d,J=8.8Hz,1H),8.33(d,J=8.4Hz,1H),7.98(d,J=7.2Hz,1H),7.93(s,1H ),7.72-7.61(m,1H),6.43(d,J=8.4Hz,1H),4.68(d,J=4.8Hz,2H),4.18-4.14(m,4H),2.18(s,2H),1.50(s,9H)ppm.

[0863] Step 3. Preparation of [2-[6-(azacyclobutane-1-yl)-2-pyridyl]-1,6-naphthid-7-yl]methylamine

[0864] TFA (1 mL) was added to a solution of N-((2-(6-(azacyclobutan-1-yl)pyridin-2-yl)-1,6-naphthidin-7-yl)methyl)carbamate (70 mg, 178.82 μmol) in DCM (3 mL) at 0 °C. The mixture was stirred at 25 °C for 2 h. The reaction mixture was poured into a saturated aqueous solution of NaHCO3 (30 mL) and extracted with EA (30 mL x 3). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to give [2-[6-(azacyclobutan-1-yl)-2-pyridinyl]-1,6-naphthidin-7-yl]methylamine (60 mg, crude) as a yellow solid, which was used for the next step without further purification.

[0865] LCMS(ESI)m / z:[M+H] + =292.4.

[0866] Step 4. Preparation of N-[[2-[6-(azacyclobutane-1-yl)-2-pyridyl]-1,6-naphthidin-7-yl]methyl]-1,1-dioxo-3,5-dihydro-2H-4,16-benzoxazothionein-8-carboxamide (1)

[0867] To a solution of 1,1-dioxo-3,5-dihydro-2H-4,1λ6-benzoxazothioneine-8-carboxylic acid (24.94 mg, 102.97 μmol) in DCM (1 mL), EDCI (21.38 mg, 111.55 μmol), HOBt (15.07 mg, 111.55 μmol), and DIEA (33.27 mg, 257.42 μmol) were added. Then, [2-[6-(azacyclobutan-1-yl)-2-pyridyl]-1,6-naphthid-7-yl]methylamine (25 mg, 85.81 μmol) was added. The mixture was stirred at 25 °C for 2 h. The reaction mixture was diluted with H2O (20 mL) and extracted with EA (30 mL x 3). The combined organic layers were washed with saline (30 mL) water, dried over Na₂SO₄, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by preparative TLC (SiO₂, DCM:MeOH = 15:1) to obtain the crude product. The crude product was then further purified by preparative HPLC (0.1% FA additive). The eluent was concentrated under reduced pressure to remove MeCN, and the residue was lyophilized to give N-[[2-[6-(azacyclobutan-1-yl)-2-pyridyl]-1,6-naphthid-7-yl]methyl]-1,1-dioxo-3,5-dihydro-2H-4,16-benzoxazothioneine-8-carboxamide (10.21 mg, 19.21 μmol, 22% yield) as a yellow solid. LCMS (ESI) m / z = [M+H] + =261.9.

[0868] 1 H NMR (400MHz, CD3OD) δ=9.33(s,1H),8.69-8.63(m,2H),8.62-8.57(m,1H),8.39(s,1H),8.24(d,J=2.0Hz,1H),7.98(s,1H),7.87(d,J=7.2Hz,1H),7 .73-7.65(m,2H),6.54(d,J=7.6Hz,1H),5.07(s,2H),4.95(s,2H),4.39- 4.34(m,2H),4.17-4.15(m,4H),3.58-3.53(m,2H),2.53-2.40(m,2H)ppm.

[0869] The following compounds in Table 2 were prepared using standard chemical operations and methods similar to those used in the preparation of Example 2.

[0870] Table 2. Compounds of the present invention

[0871]

[0872]

[0873]

[0874]

[0875]

[0876]

[0877]

[0878]

[0879] Example 3. N-[[2-[6-(azacyclobutan-1-yl)-2-pyridyl]-1,6-naphthidin-7-yl]methyl]-1,1-dioxo-3,5-dihydro-2H-4,1λ6-benzoxazothionein-8-carboxamide]

[0880]

[0881] Step 1. Preparation of (6-fluoro-2-pyridyl)-trimethyl-stanane

[0882] Trimethyl(trimethylstanyl)stanane (2.79 g, 8.52 mmol) and Pd(PPh3)4 (328.31 mg, 284.11 μmol) were added to a mixture of 2-bromo-6-fluoropyridine (500 mg, 2.84 mmol) in dioxane (5 mL). The mixture was purged with N2 for 1 min and then stirred at 100 °C for 2 h. Water (20 mL) was added, and the mixture was extracted with EtOAc (20 mL x 2). The combined organic phases were washed with brine (30 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under vacuum to give (6-fluoro-2-pyridyl)-trimethylstanane (730 mg, crude) as a brown oil. LCMS(ESI) m / z = [M + H] + =261.9.

[0883] Step 2. Preparation of N-[[2-(6-fluoro-2-pyridyl)-1,6-naphthid-7-yl]methyl]tert-butyl carbamate

[0884] Pd(PPh3)2Cl2 (71.68 mg, 102.13 μmol) was added to a mixture of N-[(2-chloro-1,6-naphthid-7-yl)methyl]carbamate (300 mg, 1.02 mmol) and 6-fluoro-2-pyridyl)-trimethyl-stanane (530.85 mg, 2.04 mmol) in dioxane (6 mL), and the mixture was purged with N2 for 1 min. The resulting mixture was stirred at 110 °C for 16 h. The reaction mixture was then poured into saturated KF (30 mL), stirred for 30 min, and extracted with EtOAc (30 mL x 2). The combined organic phases were washed with brine (40 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under vacuum. The reaction mixture (SiO2, PE:EtOAc = 20:1-1:1) was purified by column chromatography to give N-[[2-(6-fluoro-2-pyridyl)-1,6-naphthid-7-yl]methyl]carbamate tert-butyl ester (230 mg, 649.03 μmol, 64% yield) as a yellow solid.

[0885] LCMS(ESI)m / z:[M+H] + =355.1.

[0886] 1 H NMR (400MHz, CDCl3) δ = 9.26 (s, 1H), 8.63-8.58 (m, 2H), 8.41-8.39 (m, 1H), 8.04-7.98 (m, 1H), 7.94 (s, 1H), 7.09-7.06 (m, 1H), 5.49 (br s, 1H), 4.69 (br d, J = 5.2Hz, 2H), 1.50 (s, 9H) ppm.

[0887] Step 3. Preparation of [2-(6-fluoro-2-pyridyl)-1,6-naphthid-7-yl]methylamine

[0888] A mixture of N-[[2-(6-fluoro-2-pyridyl)-1,6-naphthid-7-yl]methyl]tert-butyl carbamate (220 mg, 620.81 μmol) in HCl / dioxane (2 mL) was stirred at 25 °C for 1 hr. The mixture was evaporated to dryness, and the residue was ground together with MTBE (20 mL x 2). The mixture was filtered, and the filter cake was evaporated to dryness to give [2-(6-fluoro-2-pyridyl)-1,6-naphthid-7-yl]methylamine (180 mg, crude, HCl) as a yellow solid. LCMS (ESI) m / z: [M+H] + =255.1.

[0889] Step 4. Preparation of N-[[2-(6-fluoro-2-pyridyl)-1,6-naphthid-7-yl]methyl]-1,1-dioxo-3,5-dihydro-2H-4,1λ6-benzoxazothionein-8-carboxamide]

[0890] To a mixture of [2-(6-fluoro-2-pyridyl)-1,6-naphthid-7-yl]methylamine (180 mg, 619.15 μmol) and 1,1-dioxo-3,5-dihydro-2H-4,1λ6-benzoxazothioneine-8-carboxylic acid (180 mg, 0.743 mmol) in DCM (2 mL), DIEA (320.08 mg, 2.48 mmol), EDCI (178 mg, 0.928 mmol), and HOBt (125.49 mg, 928.72 μmol) were added. The mixture was stirred at 25 °C for 1 hr. The mixture was poured into water (20 mL) and extracted with EA (10.0 mL x 3). The combined organic layers were washed with brine (20.0 mL), dried over Na2SO4, filtered, and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (0.1% FA conditions), and the eluent was concentrated under vacuum to remove MeCN. The residue was lyophilized to give N-[[2-(6-fluoro-2-pyridyl)-1,6-naphthid-7-yl]methyl]-1,1-dioxo-3,5-dihydro-2H-4,1λ6-benzoxazothionein-8-carboxamide (253 mg, 0.528 mmol, 85% yield) as a white solid.

[0891] LCMS(ESI)m / z:[M+H] + =479.0.

[0892] 1 H NMR (400MHz, DMSO-d6) δ=9.66-9.61(m,1H),9.45(d,J=0.8Hz,1H),8.75-8.72(m,1H),8.54-8.49(m,3H),8.26-8.26(m,1H),8.25-8.17(m ,1H),7.84(s,1H),7.73(d,J=8.0Hz,1H),7.39-7.36(m,1H),4.97(s,2H),4.83(d,J=5.6Hz,2H),4.23-4.21(m,2H),3.68-3.66(m,2H)ppm.

[0893] Step 5. Preparation of N-[[2-[6-[(2R)-2-methylmorpholin-4-yl]-2-pyridyl]-1,6-naphthid-7-yl]methyl]-1,1-dioxo-3,5-dihydro-2H-4,1λ6-benzoxazothionein-8-carboxamide (37)

[0894] Add DIEA (27.0 mg, 0.209 mmol) to a mixture of N-[[2-(6-fluoro-2-pyridyl)-1,6-naphthid-7-yl]methyl]-1,1-dioxo-3,5-dihydro-2H-4,1λ6-benzoxazothionein-8-carboxamide (20 mg, 0.0418 mmol) and (2R)-2-methylmorpholine hydrochloride (17.26 mg, 125.39 μmol) in DMSO (1 mL). Stir the mixture at 120 °C for 16 h. Then pour the mixture into saturated NaHCO3 (20 mL) and extract with EA (10.0 mL x 3). Wash the combined organic layers with brine (20.0 mL), dry with Na2SO4, filter, and evaporate the filtrate to dryness. The residue was purified by preparative HPLC (column: Shim-pack C18 150*25*10um; mobile phase: [water (0.225% FA)-ACN]; B%: 38%-58%, 10 min), and the eluent was concentrated under vacuum to remove MeCN. The residue was lyophilized to give N-[[2-[6-[(2R)-2-methylmorpholino-4-yl]-2-pyridyl]-1,6-naphthidin-7-yl]methyl]-1,1-dioxo-3,5-dihydro-2H-4,1λ6-benzoxazothioneine-8-carboxamide (15.89 mg, 26.24 μmol, 63% yield, FA) as a yellow solid.

[0895] LCMS(ESI)m / z:[M+H] + =560.3.

[0896] 1 H NMR (400MHz, DMSO-d6) δ = 9.65-9.62 (m, 1H), 9.40 (s, 1H), 8.68-8.61 (m, 2H), 8.54 (d, J = 2.0 Hz,1H),8.46(s,1H),8.28-8.25(m,1H),7.92(d,J=7.2Hz,1H),7.81(s,1H),7.77-7.73(m,2 H),7.03(d,J=8.4Hz,1H),4.98(s,2H),4.82(d,J=5.6Hz,2H),4.30-4.22(m,4H),3.99-3.96 (m,1H),3.70-3.58(m,4H),2.94-2.87(m,1H),2.62-2.56(m,1H),1.22(d,J=6.0Hz,3H)ppm.

[0897] The following examples in Table 3 were prepared using standard chemical operations and methods similar to those used to prepare Example 3.

[0898] Table 3. Compounds of the present invention

[0899]

[0900]

[0901]

[0902]

[0903]

[0904] Example 4. N-((2-(4-methyl-3,4-dihydro-2H-benzo[b][1,4]oxazin-8-yl)-1,6-naphthidin-7-yl)methyl)-3,5-dihydro-2H-benzo[e][1,4]oxathihepta-8-carboxamide 1,1-dioxide

[0905] Step 1: Preparation of 4-methyl-8-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-3,4-dihydro-2H-benzo[b][1,4]oxazine

[0906] Pd(dppf)Cl2 (32.1 mg, 0.0448 mmol) and AcOK (129 mg, 1.32 mmol) were added to a solution of 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-1,3,2-dioxoboronylpentane (134 mg, 0.526 mmol) and 8-bromo-4-methyl-3,4-dihydro-2H-benzo[b][1,4]oxazine (100 mg, 0.438 mmol) in dioxane (2 mL). The reaction mixture was stirred at 80 °C for 2 h. The reaction mixture was diluted with H2O (20 mL) and extracted with EA (20 mL x 3). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give the title compound (125 mg, crude) as a brown oil. LCMS (ESI) m / z: [M+H]+=276.1.

[0907] Step 2: Preparation of tert-butyl carbamate ((2-(4-methyl-3,4-dihydro-2H-benzo[b][1,4]oxazin-8-yl)-1,6-naphthid-7-yl)methyl)carbamate)

[0908] A mixture of N-[(2-chloro-1,6-naphthidin-7-yl)methyl]carbamate tert-butyl ester (100 mg, 0.340 mmol), 4-methyl-8-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-3,4-dihydro-2H-benzo[b][1,4]oxazine (122 mg, 0.443 mmol), K3PO4 (217 mg, 1.02 mmol), and [1,1′-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (22.2 mg, 0.340 mmol) in dioxane (1 mL) and H2O (0.3 mL) was degassed and purged three times with N2. The mixture was stirred at 80 °C for 2 h. The reaction mixture was diluted with H2O (10 mL) and extracted with EA (10 mL x 3). The combined organic layers were dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by reversed-phase HPLC (0.1% FA additive). The fraction was concentrated under reduced pressure to remove MeCN, and then removed with EA (50 mL x 3). The combined organic layers were dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to give the title compound (100 mg, 0.205 mmol) as a yellow solid. LCMS (ESI) m / z: [M+H]⁺ = 407.3. 1 H NMR (400MHz, DMSO-d6) δ = 9.30 (s, 1H), 8.48 (d, J = 8.8Hz, 1H), 7.98 (d, J = 8.4Hz, 1H), 7.69 (s, 1H), 7.63-7.58 (m, 1H), 7.04-7.02 (m, 1 H),6.94-6.90(m,1H),6.87-6.81(m,1H),4.43(d,J=5.6Hz,2H),4.35-4.27(m,2H),3.34-3.33(m,2H),2.91(s,3H),1.43(s,9H)ppm.

[0909] Step 3: Preparation of (2-(4-methyl-3,4-dihydro-2H-benzo[b][1,4]oxazin-8-yl)-1,6-naphthid-7-yl)methylamine hydrochloride

[0910] HCl / dioxane (4N, 750 μL) was added to a solution of ((2-(4-methyl-3,4-dihydro-2H-benzo[b][1,4]oxazin-8-yl)-1,6-naphthid-7-yl)methyl)carbamate tert-butyl ester (90 mg, 0.221 mmol) in dioxane (1 mL). The reaction mixture was stirred at 25 °C for 1 hr. The reaction mixture was concentrated under reduced pressure. The residue was washed with MTBE (5 mL x 2), filtered, and dried under vacuum to give the title compound (70 mg, 0.204 mmol) as a brown solid. LCMS (ESI) m / z: [M+H]+ = 307.2.

[0911] Step 4: Preparation of N-((2-(4-methyl-3,4-dihydro-2H-benzo[b][1,4]oxazin-8-yl)-1,6-naphthidin-7-yl)methyl)-3,5-dihydro-2H-benzo[e][1,4]oxathihepta-8-carboxamide 1,1-dioxide (58)

[0912] EDCI (25.2 mg, 0.131 mol), HOBt (17.7 mg, 0.131 mmol), DIEA (76.2 μL, 0.438 mmol), and (2-(4-methyl-3,4-dihydro-2H-benzo[b][1,4]oxazin-8-yl)-1,6-naphthid-7-yl)methylamine hydrochloride (30 mg, 0.0875 mmol) were added to a solution of 2,3-dihydro-5H-benzo[e][1,4]oxathiaheptaphylline-8-carboxylic acid 1,1-dioxide (25.4 mg, 0.105 mmol) in DCM (0.5 mL). The reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was diluted with H2O (5 mL) and extracted with DCM (5 mL x 3). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by reversed-phase HPLC (0.1% FA conditions). The solution was concentrated under reduced pressure to remove MeCN, and lyophilized to give the title compound (14.2 mg, 0.0257 mmol) as a yellow solid. LCMS (ESI) m / z: [M+H]+ = 531.2. 1H NMR (400MHz, CD3OD) δ = 9.30 (s, 1H), 8.64-8.59 (m, 1H), 8.48 (d, J = 8.8Hz, 1H ),8.22-8.20(m,1H),8.00(d,J=8.4Hz,1H),7.93(s,1H),7.63(d,J=7.6Hz,1 H),7.01-6.92(m,2H),6.88-6.83(m,1H),5.04(s,2H),4.92(s,2H),4.35-4 .31(m,4H),3.53-3.50(m,2H),3.34(d,J=4.4Hz,2H),2.97-2.92(m,3H)ppm.

[0913] The following examples in Table 4A were prepared using standard chemical operations and methods similar to those used in the preparation of Example 4.

[0914] Table 4A. Compounds of the present invention

[0915]

[0916]

[0917]

[0918]

[0919]

[0920] The following examples in Table 4B were prepared using standard chemical procedures and methods similar to those described above.

[0921] Table 4B. Compounds of the present invention

[0922]

[0923]

[0924]

[0925]

[0926]

[0927]

[0928]

[0929]

[0930]

[0931]

[0932]

[0933]

[0934]

[0935]

[0936]

[0937]

[0938]

[0939]

[0940]

[0941]

[0942]

[0943]

[0944]

[0945]

[0946]

[0947]

[0948]

[0949]

[0950]

[0951]

[0952]

[0953]

[0954]

[0955]

[0956]

[0957]

[0958]

[0959]

[0960]

[0961]

[0962]

[0963]

[0964]

[0965]

[0966]

[0967]

[0968]

[0969]

[0970]

[0971]

[0972]

[0973]

[0974]

[0975]

[0976]

[0977]

[0978]

[0979]

[0980]

[0981]

[0982]

[0983]

[0984]

[0985]

[0986]

[0987]

[0988]

[0989]

[0990]

[0991]

[0992]

[0993]

[0994]

[0995]

[0996]

[0997]

[0998]

[0999]

[1000]

[1001]

[1002]

[1003]

[1004]

[1005]

[1006]

[1007]

[1008]

[1009]

[1010]

[1011] Example 5. Determination of ATPase catalytic activity of BRM and BRG-1

[1012] By using ADP-Glo TM (Promega, V9102) was used in vitro biochemical assays to measure the ATPase catalytic activity of BRM or BRG-1. Once the reaction was complete, ADP-Glo ​​was performed in two steps. TM Kinase assay. The first step is to deplete any unused ATP in the reaction system. The second step is to convert the reaction product ADP into ATP, which will be used by luciferase to produce light and be detected by a luminescent reader such as Envision.

[1013] The assay reaction mixture (10 μL) contains 30 nM BRM or BRG-1 and 20 nM salmon sperm DNA (from Invitrogen, UltraPure). TM Salmon Sperm DNA Solution (catalog number 15632011) and 400 μM ATP in an ATPase assay buffer containing 20 mM Tris, pH 8, 20 mM MgCl2, 50 mM NaCl, 0.1% Tween-20, and 1 mM fresh DTT (Pierce) TM DTT (dithiothreitol, catalog number 20290). The reaction was initiated by adding 2.5 μL of ATPase solution to 2.5 μL of ATP / DNA solution on a low-volume white Proxiplate-384+ plate (PerkinElmer, catalog number 6008280) and incubating at room temperature for 1 hour. Then, 5 μL of ADP-Glo ​​provided in the kit was added. TM After adding the reagents, the reaction system was incubated at room temperature for 40 minutes. Then, 10 μL of the kinase assay reagent provided in the kit was added to convert ADP to ATP, and the reaction system was incubated at room temperature for 60 minutes. Finally, luminescence measurements were collected using a plate-reading photometer such as Envision.

[1014] BRM and BRG-1 were synthesized from the high five insect cell line with a purity greater than 90%. The IC50 values ​​from the ATPase catalytic activity assay described herein... 50 The data is shown in Tables 5A and 5B below.

[1015] Table 5A. BRM and BRG-1 inhibition data of the compounds of the present invention

[1016]

[1017]

[1018]

[1019]

[1020] *Ratio is BRG1 IC 50 (μM) divided by BRM IC 50 The value generated by (μM).

[1021] Table 5B. BRM and BRG-1 inhibition data of the compounds of the present invention

[1022]

[1023]

[1024]

[1025]

[1026]

[1027]

[1028]

[1029] Example 6. Synthesis of Compound A

[1030] BRG1 / BRM inhibitor compound A has the following structure:

[1031]

[1032] Compound A is synthesized as shown in Scheme 1 below.

[1033] Scheme 1. Synthesis of Compound A

[1034]

[1035] By using the above ADP-Glo TM An in vitro biochemical assay using Promega (V9102) was used to determine the ATPase catalytic activity of BRM or BRG-1 in the presence of compound A. Compound A was found to have an IC50 of 10.4 nM for BRM in this assay. 50IC with 19.3nM for BRG1 50 .

[1036] Example 7. Effects of BRG1 / BRM ATPase inhibition on the growth of ocular melanoma and hematologic malignancies cell lines.

[1037] Methods: Ocular melanoma cell lines (92-1, MP41, MP38, MP46), prostate cancer cell line (LNCAP), lung cancer cell line (NCI-H1299), and indefinitely proliferating embryonic kidney cell line (HEK293T) were plated in 96-well plates containing growth medium (see Table 6). BRG1 / BRM ATPase inhibitor compound A was dissolved in DMSO and added to the cells at a concentration gradient of 0 to 10 μmol during plate formation. Cells were incubated at 37°C for 3 days. After 3 days of treatment, the culture medium was removed from the cells, and 30 μL of TrypLE (Gibco) was added to the cells for 10 minutes. The cells were then separated from the plate and resuspended by adding 170 μL of growth medium. Cells from the two DMSO-treated control wells were counted, and the initial cell count was replated at 37°C in plates containing fresh compound for 4 days. On day 7, cells were harvested as described above. Relative cell growth was measured on days 3 and 7 by adding Cell-titer glo (Promega), and luminescence was measured on an Envision plate reader (Perkin Elmer). The concentration at which growth was inhibited by 50% for each cell line was calculated using GraphpadPrism. 50 ), and plotted as follows. For multiple myeloma cell lines (OPM2, MM1S, LP1), ALL cell lines (TALL1, JURKAT, RS411), DLBCL cell lines (SUDHL6, SUDHL4, DB, WSUDLCL2, PFEIFFER), AML cell lines (OCIAML5), MDS cell lines (SKM1), ovarian cancer cell lines (OV7, TYKNU), esophageal cancer cell lines (KYSE150), rhabdomyosarcoma cell lines (RD, G402, G401, HS729, A204), liver cancer cell lines (HLF, HLE, PLCRPF5), and lung cancer cell lines (SW1573, NCIH2444), the above method can be implemented with the following modifications: cells are plated in 96-well plates, and the next day, BRG1 / BRM ATPase inhibitor compound A is dissolved in DMSO and added to the cells in a concentration gradient from 0 to 10 μmol. During cell division on days 3 and 7, cells were transferred into new 96-well plates, and fresh compound was added 4 hours after repotting.

[1038] Table 6 lists the cell lines tested and the growth media used.

[1039] Table 6. Cell lines and growth media

[1040] cell lines source growth medium 92-1 SIGMA RPMI 1640 + 20% FBS A204 ATCC McCoy's 5A+ 10% FBS DB ATCC RPMI 1640 + 10% FBS G401 ATCC McCoy's 5A+ 10% FBS G402 ATCC McCoy's 5A+ 10% FBS HEK293T ATCC DMEM + 10% FBS HLE JCRB DMEM + 10% FBS HLF JCRB DMEM + 10% FBS HS729 ATCC DMEM + 10% FBS JURKAT ATCC RPMI 1640 + 10% FBS KYSE150 DSMZ RPMI1640 / Ham's F12 + 10% FBS LNCAP ATCC RPMI 1640 + 10% FBS LP1 DSMZ IMDM + 20% FBS MM1S ATCC RPMI 1640 + 10% FBS MP38 ATCC RPMI 1640 + 20% FBS MP41 ATCC RPMI 1640 + 20% FBS MP46 ATCC RPMI 1640 + 20% FBS NCIH1299 ATCC RPMI 1640 + 10% FBS NCIH2444 ATCC RPMI 1640 + 20% FBS OCIAML5 DSMZ alpha-MEM+20%FBS+10ng / mlGM-CSF OPM2 DSMZ RPMI 1640 + 10% FBS OV7 ECACC DMEM / Ham's F12 (1:1) + 2mM glutamine + 10% FBS + 0.5ug / ml hydrocortisone + 10ug / ml insulin PFEIFFER ATCC RPMI 1640 + 10% FBS PLCPRF5 ATCC EMEM + 10% FBS RD ATCC DMEM + 10% FBS RS411 ATCC RPMI 1640 + 10% FBS SKM1 JCRB RPMI 1640 + 10% FBS SUDHL4 DSMZ RPMI 1640 + 10% FBS SUDHL6 ATCC RPMI 1640 + 20% FBS SW1573 ATCC DMEM + 10% FBS TALL1 JCRB RPMI 1640 + 10% FBS TYKNU JCRB EMEM + 20% FBS WSUDLCL2 DSMZ RPMI 1640 + 10% FBS

[1041] Result: As Figure 1 As shown, ocular melanoma and hematologic malignancies cell lines were more sensitive to BRG1 / BRM inhibition than other tested cell lines. Inhibition against ocular melanoma and hematologic malignancies cell lines was maintained until day 7.

[1042] Example 8. Comparison of BRG1 / BRM inhibitors with clinical PKC and MEK inhibitors in ocular lentigines melanoma cell lines.

[1043] Methods: Ocular melanoma cell lines 92-1 or MP41 were plated in 96-well plates in the presence of growth medium (see Table 5). BAF ATPase inhibitors (compound A), PKC inhibitors (LXS196; MedChemExpress), or MEK inhibitors (selutinib; Selleck Chemicals) were dissolved in DMSO and added to the cells at concentration gradients from 0 to 10 μmol during plate formation. Cells were incubated at 37°C for 3 days. After 3 days of treatment, cell growth was measured using a promega luminescence assay, and the luminescence was read on an Envision plate reader (Perkin Elmer).

[1044] Result: As Figure 2A and Figure 2B As shown, compound A exhibited growth inhibition comparable to clinical PKC and MEK inhibitors against ocular retinal melanoma cells. Furthermore, compound A was found to produce an inhibitory effect with a faster onset of action than clinical PKC and MEK inhibitors.

[1045] Example 9. Synthesis of Compound B

[1046] BRG1 / BRM inhibitor compound B has the following structure:

[1047]

[1048] The following two schemes show the synthesis of compound B.

[1049] Option 2: Synthesis of Compound B

[1050]

[1051] Preparation of (S)-1-(methylsulfonyl)-N-(4-(methylthio)-1-oxo-1-((4-(3-(pyridin-4-yl)phenyl)thiazo-2-yl)amino)but-2-yl)-1H-pyrrole-3-carboxylate (compound B)

[1052]

[1053] Add EDCI (1.37 g, 7.13 mmol), HOBt (962.92 mg, 7.13 mmol), and DIEA (2.46 g, 19.00 mmol, 3.31 mL) to a mixture of (2S)-2-amino-4-methylthioalkyl-N-[4-[3-(4-pyridyl)phenyl]thiazolyl-2-yl]butyramide (2 g, 4.75 mmol, HCl salt) and 1-methylsulfonylpyrrole-3-carboxylic acid (898.81 mg, 4.75 mmol) in DMF (20 mL), and stir the mixture at 25 °C for 3 hours. Pour the mixture into H2O (100 mL) and collect the precipitate by filtration. Grind the solid in MeOH (20 mL) and collect the precipitate by filtration. The solid was dissolved in DMSO (10 mL), and the mixture was then poured into MeOH (50 mL). The precipitate was collected by filtration and lyophilized to give compound B (2.05 g, 3.66 mmol, 77.01% yield) as a white solid. LCMS (ESI) m / z [M+H] + =555.9. 1 H NMR (400MHz, DMSO) δ12.49 (s, 1H), 8.68-8.66 (m, 2H), 8.46 (d, J = 7.2Hz, 1H), 8.31-8.30 (m ,1H),8.02-8.00(m,1H),7.94-7.96(m,1H),7.83(s,1H),7.73-7.74(m,3H),7.61-7.57(m ,1H),7.31-7.29(m,1H),6.79-6.77(m,1H),4.74-4.69(m,1H),3.57(s,3H),2.67-2.53(m ,2H),2.13-2.01(m,5H).SFC:AS-3-MeOH(DEA)-40-3mL-35T.lcm,t=0.932min,ee%=100%.

[1054] Example 10. Effects of BRG1 / BRM ATPase inhibition on the growth of ocular melanoma, hematologic malignancies, prostate cancer, breast cancer, and Ewing sarcoma cell lines.

[1055] Methods: All cell lines described in Example 7 above were tested with compound B as described above. In addition, the following cell lines were also tested: Ewing sarcoma cell lines (CADOES1, RDES, SKES1), retinoblastoma cell lines (WERIRB1), ALL cell lines (REH), AML cell lines (KASUMI1), prostate cancer cell lines (PC3, DU145, 22RV1), melanoma cell lines (SH4, SKMEL28, WM115, COLO829, SKMEL3, A375), and breast cancer cell lines (MDAMB415, CAMA1, MCF7, BT). The following methods were modified for the use of BRG1 / BRM ATPase inhibitor compound B in 96-well plates: 474, HCC1419, DU4475, BT549, B-ALL cell line (SUPB15), CML cell line (K562, MEG01), Burkitt lymphoma cell line (RAMOS2G64C10, DAUDI), mantle cell lymphoma cell line (JEKO1, REC1), bladder cancer cell line (HT1197), and lung cancer cell line (SBC5). Cells were seeded in 96-well plates. On the second day, BRG1 / BRM ATPase inhibitor compound B was dissolved in DMSO and added to the cells at concentration gradients from 0 to 10 μmol. On days 3 and 7, cells were divided and transferred to new 96-well plates, with fresh compound added 4 hours after reseeding.

[1056] Table 7 lists the cell lines tested and the growth media used.

[1057] Table 7. Cell lines and growth media

[1058] cell lines source growth medium 22RV1 ATCC RPMI 1640 + 10% FBS A375 ATCC DMEM + 10% FBS BT474 ATCC Hybricare medium + 1.5g / L sodium bicarbonate + 10% FBS BT549 ATCC RPMI 1640 + 0.023 IU / ml insulin + 10% FBS CADOES1 DSMZ RPMI 1640 + 10% FBS CAMA1 ATCC EMEM + 10% FBS COLO829 ATCC RPMI 1640 + 10% FBS DAUDI ATCC RPMI 1640 + 10% FBS DU145 ATCC EMEM + 10% FBS DU4475 ATCC RPMI 1640 + 10% FBS HCC1419 ATCC RPMI 1640 + 10% FBS HT1197 ATCC EMEM + 10% FBS JEKO1 ATCC RPMI 1640 + 20% FBS K562 ATCC IMDM + 10% FBS KASUMI1 ATCC RPMI 1640 + 10% FBS MCF7 ATCC EMEM + 0.01 mg / ml bovine insulin + 10% FBS MDAMB415 ATCC Leibovitz's L-15 + 2mM L-glutamine + 10mcg / ml insulin + 10mg / ml glutathione + 15% FBS MEG01 ATCC RPMI 1640 + 10% FBS PC3 ATCC F-12K+10% FBS RAMOS2G64C10 ATCC RPMI 1640 + 10% FBS RDES ATCC RPMI 1640 + 15% FBS REC1 ATCC RPMI 1640 + 10% FBS REH ATCC RPMI 1640 + 10% FBS SBC5 JCRB EMEM + 10% FBS SH4 ATCC DMEM + 10% FBS SKES1 ATCC McCoy's 5A+15% FBS SKMEL28 ATCC EMEM + 10% FBS SKMEL3 ATCC McCoy's 5A+15% FBS SUPB15 ATCC IMDM + 4mM L-glutamine + 1.5g / L sodium bicarbonate + 0.05mM 2-mercaptoethanol + 20% FBS WERIRB1 ATCC RPMI 1640 + 10% FBS WM115 ATCC EMEM + 10% FBS

[1059] Result: As Figure 3 As shown, ocular melanoma, hematologic malignancies, prostate cancer, breast cancer, and Ewing sarcoma cell lines were more sensitive to BRG1 / BRM inhibition than other tested cell lines. The inhibitory effect on ocular melanoma, hematologic malignancies, prostate cancer, breast cancer, and Ewing sarcoma cell lines persisted until day 7.

[1060] Example 11. Effect of BRG1 / BRM ATPase inhibition on cancer cell line growth

[1061] Methods: As previously described (“High-throughput identification of genotype-specific cancer vulnerabilities in mixtures of barcoded tumor cell lines”, Yu et al., Nature Biotechnology 34, 419-423, 2016), pooled cell viability assays were performed using the following modifications. Cell lines were obtained from the Cancer Cell Line Encyclopedia (CCLE) collection and adapted to phenol red-free RPMI-1640 medium supplemented with 10% heat-inactivated fetal bovine serum (FBS) to allow for a unique infection and pooling protocol applicable to such a large range of cell lines. A lentiviral rotation infection protocol was performed to introduce a 24-nucleotide barcode in each cell line, using blasticidin as a selection marker, with an estimated fold increase (MOI) of 1 for all cell lines. Over 750 stably barcoded PRISM cancer cell lines were then pooled together according to doubling time, 25 per pool. For screening, instead of using the previously described (Yu et al.) method of plate-capping 25 cell lines in each well, all adherent cells or all suspension cell lines were plated together in T25 flasks (100,000 cells / flask) or 6-well plates (50,000 cells / well). Cells were treated with DMSO or a compound at a maximum concentration of 10 μM, in triplicate at 8 3-fold dose responses. As controls for stability assessment, cells were treated in parallel with two previously validated compounds, the pan-Raf inhibitor AZ-628 and the proteasome inhibitor bortezomib, at maximum concentrations of 2.5 μM and 0.039 μM, respectively.

[1062] After treatment with the compound for 3 days, cells were lysed, genomic DNA was extracted, barcodes were amplified by PCR, and detected by Next-Generation Sequencing. Cell viability was determined by comparing the count of cell line-specific barcodes in the treated samples with the counts in the DMSO control and the day 0 control. Dose-response curves were fitted for each cell line, and the corresponding area under the curve (AUC) was calculated and compared with the median AUC of all cell lines. Figure 4 Cell lines with an AUC less than the median value are considered the most sensitive.

[1063] Example 12. Effects of BRG1 / BRM ATPase inhibitors on ocular uveal melanoma cell lines.

[1064] Methods: Ocular melanoma cell lines (92-1, MP41, MP38, MP46) and non-small cell lung cancer cells (NCIH1299) were plated in 96-well plates containing growth medium (see Table 6). BRG1 / BRM ATPase inhibitor compound 67 was dissolved in DMSO and added to the cells at concentration gradients from 0 to 10 μmol during plate formation. Cells were incubated at 37°C for 3 days. After 3 days of treatment, cell growth was measured using a promega luminescence assay, and the luminescence was read on an Envision plate reader (Perkin Elmer).

[1065] Result: As Figure 5 As shown, compound B resulted in effective growth inhibition in ocular pigment melanoma cell lines.

[1066] Example 13. Comparison of BRG1 / BRM inhibitors with clinical PKC and MEK inhibitors in ocular uveal melanoma cells.

[1067] Methods: Ocular melanoma cell lines 92-1 or MP41 were plated in 96-well plates in the presence of growth medium (see Table 6). BAF ATPase inhibitor (compound B), PKC inhibitor (LXS196; MedChemExpress), and MEK inhibitor (selmetinib; Selleck Chemicals) were dissolved in DMSO and added to the cells at concentration gradients from 0 to 10 μmol during plate formation. Cells were incubated at 37°C for 3 days. After 3 days of treatment, cell growth was measured using a promega luminescence assay, and the luminescence was read on an Envision plate reader (Perkin Elmer).

[1068] Result: As Figure 6A and Figure 6B As shown, compound B exhibited a more potent inhibitory effect on the growth of ocular lentigines melanoma cells compared to clinical PKC and MEK inhibitors. Furthermore, compound B was found to initiate growth inhibition more rapidly than clinical PKC and MEK inhibitors.

[1069] Example 14. BRG1 / BRM ATPase inhibitors effectively inhibit the growth of PKC inhibitor-resistant cells.

[1070] Methods: MP41 ocular retinal melanoma cells were induced to develop resistance to the PKC inhibitor (LXS196; MedChemExpress) by long-term culture in a growth medium containing incremental concentrations of up to 1 μmol of the compound (see Table 6). After 3 months, the sensitivity of parental MP41 cells and PKC inhibitor (PKCi)-resistant cells to either the PKC inhibitor (LXS196) or the BRG1 / BRM ATPase inhibitor (compound B) was tested in a 7-day growth inhibition assay as described in Example 9 above.

[1071] Results: Although PKCi resistant cells could grow more tolerantly to higher concentrations of LXS196 than the parental MP41 cell line ( Figure 7A However, the BRG1 / BRM ATPase inhibitor (compound B) still resulted in strong growth inhibition of PKCi resistance and parental cell lines. Figure 7B PKCi resistant cells are more sensitive to compound B than parental MP41 cells. Figure 7B ).

[1072] Example 15. Synthesis of Compound C

[1073]

[1074] Step 1. Preparation of 6-fluoropyridine-2-carbonyl chloride (intermediate B)

[1075]

[1076] Oxaloyl chloride (155.10 mL, 1.77 mol) was added to a solution of cooled (0 °C) 6-fluoropyridine-2-carboxylic acid (50.00 g, 354.36 mmol) in dichloromethane (500 mL) and N,N-dimethylformamide (0.26 mL, 3.54 mmol). After the addition of oxalloyl chloride was complete, the reaction mixture was warmed to room temperature and stirred for 0.5 h. The mixture was concentrated under vacuum to give intermediate B (56.50 g) as a white solid, which was used for the next step without further purification.

[1077] Step 2. Preparation of 2-chloro-1-(6-fluoro-2-pyridyl)acetone (intermediate C)

[1078]

[1079] A 2M trimethylsilyldiazomethane solution in hexane (351 mL) was added dropwise to a mixture of cooled (0 °C) intermediate B (56.00 g, 351.00 mmol) in 1,4-dioxane (800 mL). The resulting reaction mixture was stirred at 25 °C for 10 h. The reaction mixture was then quenched with a 4M HCl solution in 1,4-dioxane (500 mL). After stirring for 2 h, the reaction solution was concentrated under vacuum to give an oily substance. The residue was diluted with a saturated aqueous solution of NaHCO3 (500 mL) and extracted with ethyl acetate (200 mL x 3). The combined organic layers were washed with brine (300 mL x 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to give intermediate C (35.50 g) as a white solid, which was used directly in the next step. LCMS (ESI) m / z: [M+H] + =173.8.

[1080] Step 3.4 Preparation of (6-fluoro-2-pyridyl)thiazole-2-amine (intermediate E)

[1081]

[1082] NaF (3.56 g, 84.82 mmol) was added to a solution of intermediate C (35.50 g, 204.53 mmol) and thiourea (14.01 g, 184.07 mmol) in a mixture of MeOH (250 mL) and H₂O (250 mL) at room temperature. After stirring for 0.5 h, the reaction mixture was partially concentrated under vacuum to remove MeOH, and the resulting solution was acidified to pH ~3 with 2 M HCl aqueous solution. After 15 min, the solution was extracted with ethyl acetate (200 mL x 3), the organic layer was discarded, the aqueous phase was alkalized with NaHCO₃ (500 mL), stirred for 30 min, and then extracted with ethyl acetate (325 mL x 3). The combined organic layers were washed with brine (225 mL x 3), dried with Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was ground with petroleum ether (300 mL), stirred at 25 °C for 10 min, and filtered. The solid obtained by vacuum drying yielded intermediate E (28.00 g, 143.43 mmol, 70.13% yield, 100% purity), a white solid. LCMS (ESI) m / z: [M+H] + =195.8; 1 H NMR (400MHz, DMSO-d6) δ8.00-7.96 (m, 1H), 7.72 (d, J = 7.2Hz, 1H), 7.24 (s, 1H), 7.16 (s, 2H), 7.02 (d, J = 8.0Hz, 1H).

[1083] Step 4. Preparation of N-[2-[[4-(6-fluoro-2-pyridyl)thiazolyl-2-yl]amino]-2-oxo-ethyl]tert-butyl carbamate (intermediate G)

[1084]

[1085] Intermediate E (6.00 g, 30.74 mmol) was added to a solution of N-Boc-glycine (5.92 g, 33.81 mmol), HATU (12.86 g, 33.81 mmol), and DIEA (15.89 g, 122.94 mmol, 21.41 mL) in dichloromethane (100 mL). After stirring for 2 h, the reaction mixture was concentrated, diluted with water (100 mL), and extracted with ethyl acetate (60 mL x 4). The combined organic layers were washed with brine (100 mL x 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was ground together with a 1:1 mixture of petroleum ether and MeOH (40 mL). After stirring at 25°C for 20 min, the suspension was filtered, the filter cake was washed with MTBE (20 mL), and dried under vacuum to give intermediate G (7.7 g, 21.63 mmol, 70.4% yield, 99.0% purity), as a white solid. LCMS (ESI) m / z: [M+H] + =353.1.

[1086] Step 5. Preparation of 2-((4-(6-fluoropyridin-2-yl)thiazolyl)amino)-2-oxoethyl-1-ammonium chloride (intermediate H)

[1087]

[1088] The solution of intermediate G (5.40 g, 15.32 mmol) in 35 mL of 4 M HCl in 1,4-dioxane was stirred at 25 °C for 1.5 h. The mixture was then concentrated under vacuum to give intermediate H (4.42 g) as a white solid, which was used directly for the next step without further purification. LCMS (ESI) m / z: [M+H] + =252.9.

[1089] Step 6. Preparation of 1-tert-butyl-N-[2-[[4-(6-fluoro-2-pyridyl)thiazolyl-2-yl]amino]-2-oxo-ethyl]pyrrole-3-carboxamide (intermediate J)

[1090]

[1091] HOBt (1.68 g, 12.47 mmol), EDCI (2.39 g, 12.47 mmol), and DIEA (6.71 g, 51.95 mmol, 9.05 mL) were added sequentially to a solution of intermediate H (3.00 g, 10.39 mmol), 1-tert-butylpyrrole-3-carboxylic acid (1.74 g, 10.39 mmol), and DIEA (6.71 g, 51.95 mmol, 9.05 mL) in dichloromethane (40 mL). After stirring for 4 h, the mixture was concentrated under vacuum. The residue was diluted with water (250 mL) and extracted with ethyl acetate (200 mL x 3). The combined organic layers were washed with brine (300 mL x 3), dried over Na2SO4, filtered, and concentrated under reduced pressure. The resulting solid was ground together with a 1:1 MTBE / ethyl acetate mixture (400 mL), and the suspension was filtered after 30 min. The solid was washed with MTBE (85 mL x 3) and then dried under vacuum to give intermediate J (3.10 g, 7.64 mmol, 73.6% yield, 99.0% purity) as a white solid. LCMS (ESI) m / z: [M+H] + =402.3. 1 H NMR(400MHz,DMSO-d6)δ12.40(s,1H),8.18-8.15(m,1H),8.09-8.08(m,1H),7.87-7.83(m,2H),7 .52(s,1H),7.11(d,J=8.0Hz,1H),6.97(m,1H),6.47(s,1H),4.10(d,J=5.6Hz,2H),1.49(s,9H).

[1092] Step 7. Preparation of 1-(tert-butyl)-N-(2-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazo-2-yl)amino)-2-oxoethyl)-1H-pyrrole-3-carboxylate (compound C)

[1093]

[1094] To a solution of intermediate J (0.100 g, 0.249 mmol) in DMSO (1 mL), DIEA (0.130 mL, 0.747 mmol) and cis-2,6-dimethylmorpholine (0.057 g, 0.498 mmol) were added, and the mixture was stirred at 120 °C. After 12 h, the solution was cooled to room temperature, and the reaction mixture was diluted with MeOH (3 mL). The residue was purified by preparative HPLC (0.1% TFA; column: Luna C18 150*25 5 u; mobile phase: [water (0.075% TFA)-ACN]; B%: 30%-60%, 2 min). Suitable fractions were collected, lyophilized, and compound C (0.079 g, 0.129 mmol, 51.94% yield, 100% purity) was given as a white solid. LCMS (ESI) m / z: [M+H] + =497.5;

[1095] 1 H NMR(400MHz,DMSO-d6)δ12.27(s,1H),8.17-8.14(m,1H),7.75(s,1H),7. 63-7.59(m,1H),7.51(s,1H),7.25(d,J=7.2Hz,1H),6.96(s,1H),6.79(d, J=8.8Hz,1H),6.47(s,1H),4.24(d,J=12.4Hz,2H),4.08(d,J=5.6Hz,2H), 3.64-3.61(m,2H),2.44-2.38(m,2H),1.49(s,9H),1.18(d,J=5.6Hz,6H).

[1096] Example 16. BRG1 / BRM ATPase inhibitors induced growth inhibition of ocular retinal melanoma in vivo.

[1097] Methods: 5x10 cm² subcutaneous implants were placed in the axillary region of nude mice (Envigo). 6 92-1 melanoma cells from the retina pigmentosa. This caused the tumor to grow to approximately 200 mm. 3 At this point, mice were grouped and administration began. Mice were administered the medium (20% 2-hydroxypropyl-β-cyclodextrin) or escalating doses of compound C once daily via oral tube feeding. Tumor volume and body weight were measured over 3 weeks, and the dose was adjusted accordingly to achieve an appropriate dose in mg / kg. At this point, the animals were euthanized, the tumors were dissected, and imaging was performed.

[1098] Results: Compound C induced tumor growth inhibition in a dose-dependent manner, with tumor regression observed at the highest dose (50 mg / kg). Figure 8A and Figure 8B All treatments were well tolerated, and no weight loss was observed. Figure 8C ).

[1099] Other implementation plans

[1100] Although the invention has been described in conjunction with specific embodiments thereof, it should be understood that the invention is capable of further modifications, and this application is intended to cover any variations, uses, or alterations to the principles generally followed by the invention and to include such deviations from the disclosure within the scope of known or customary practice in the field to which the invention pertains, which may apply to the basic features described above and within the scope of the claims.

[1101] Other embodiments are described in the claims.

Claims

1. Compounds having the following structure: in m is 1; n is 0, 1, 2, 3 or 4; R 1 It is hydrogen or C1-C6 alkyl; Each R 2 and each R 3 Independently hydrogen or C1-C6 alkyl, wherein the alkyl group is unsubstituted or substituted with 1 to 4 substituents, the substituents being independently selected from C1-C6 alkyl, C6 ... 10 Aryl-C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxyalkyl, phenyl, C3-C6 cycloalkyl, halogen, hydroxyl, methoxy, ethoxy, C1-C6 thioalkoxy, 5-10 membered heteroaryl, 4-12 membered heterocyclic, NH2, or mono- or bis-C1-C6 alkylamino, azide, cyano, nitro and thiol; L 1 for in X 3 X 4 X 5 X 6 X 7 and X 8 Each is independently N or CR L1 ; Each R L1 Independently, it can be H, halogen, or C1-C6 alkyl. A 1 It is bonded to –(C(R) 2 (R) 3 )) m - the key; and A 2 Is it bonded to L 2 The key; L 2 It does not exist; it is C3-C. 10 cycloalkyl, C6-C 10 Aryl, 5- to 14-membered heteroaryl, or 4- to 14-membered heterocyclic, wherein the cycloalkyl, aryl, heteroaryl, or heterocyclic group is independently unsubstituted or substituted by 1 to 4 substituents, wherein the substituents are independently selected from C1-C6 alkyl, C6 ... 10 Aryl-C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxyalkyl, phenyl, C3-C6 cycloalkyl, halogen, hydroxyl, methoxy, ethoxy, C1-C6 thioalkoxy, 5-10 heteroaryl, 4-12 membered heterocyclic, NH2, or mono- or bis-C1-C6 alkylamino, azide, cyano, nitro and thiol; in Groups with the following structures in Z represents CH2 or CO; Each R X1 Independently C1-C6 alkyl or halogen, or two twin Rs X1 Groups and the atoms they are attached to combine to form carbonyl groups; p is 0, 1, 2, 3 or 4; It can be a single bond or a double bond; R 4 It is hydrogen, halogen, C1-C6 alkyl or C3-C 10 cycloalkyl; Each R 7 Independently C1-C6 alkyl, C1-C6 heteroalkyl, halogen, C3-C 10 cycloalkyl, C3-C 10 Cycloalkyl C1-C6 alkyl, 5- to 14-membered heteroaryl, 4- to 14-membered heterocyclic, -N(R) 7A )2 or -OR 7A The alkyl, heteroalkyl, cycloalkyl, heteroaryl, or heterocyclic groups therein are each independently unsubstituted or substituted by 1 to 4 substituents, wherein the substituents are independently selected from C1-C6 alkyl, C6 ... 10 Aryl-C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxyalkyl, phenyl, C3-C6 cycloalkyl, halogen, hydroxyl, methoxy, ethoxy, C1-C6 thioalkoxy, 5-10 heteroaryl, 4-12 membered heterocyclic, NH2, or mono- or bis-C1-C6 alkylamino, azide, cyano, nitro and thiol; Each R 7A Independently H, or C1-C6 alkyl, C3-C 10 cycloalkyl, C6-C 10 Aryl, 5- to 10-membered heteroaryl, or 4- to 10-membered heterocyclic, wherein the alkyl, cycloalkyl, aryl, heteroaryl, or heterocyclic group is independently unsubstituted or substituted by 1 to 4 substituents, wherein the substituents are independently selected from C1-C6 alkyl, C6 ... 10 aryl-C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxyalkyl, phenyl, C3-C6 cycloalkyl, halogen, hydroxyl, methoxy, ethoxy, C1-C6 thioalkoxy, 5-10 heteroaryl, 4-12 membered heterocyclic, NH2, or mono- or bis-C1-C6 alkylamino, azide, cyano, nitro, and thiol; or two twin R 7 The group and the attached atom combine to form a carbonyl group; R 8 It is hydrogen, halogen, C1-C6 alkyl or C3-C 10 Cycloalkyl, wherein each of the alkyl or cycloalkyl groups is independently unsubstituted or substituted by 1 to 4 substituents, wherein the substituents are independently selected from C1-C6 alkyl, C6 ... 10 Aryl-C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxyalkyl, phenyl, C3-C6 cycloalkyl, halogen, hydroxyl, methoxy, ethoxy, C1-C6 thioalkoxy, 5-10 heteroaryl, 4-12 membered heterocyclic, NH2, or mono- or bis-C1-C6 alkylamino, azide, cyano, nitro, and thiol; and R 9 It is hydrogen; The heteroaryl group comprises 1, 2 or 3 ring atoms selected from nitrogen, oxygen and sulfur, and the heterocyclic group comprises 1, 2, 3 or 4 ring atoms selected from nitrogen, oxygen and sulfur; Or its pharmaceutically acceptable salt.

2. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R 8 It is hydrogen or halogen.

3. The compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein at least one R X1 It is a C1-C6 alkyl group, or at least one R X1 It is a halogen, or at least two of its twins R X1 The group and the atom it is attached to combine to form a carbonyl group.

4. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein L 1 for 5. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein -L 2 -(R 7 ) n Groups with the following structures:

6. The compound of claim 5 or a pharmaceutically acceptable salt thereof, wherein -L 2 -(R 7 ) n Groups with the following structures:

7. The compound of claim 5 or a pharmaceutically acceptable salt thereof, wherein -L 2 -(R 7 ) n Groups with the following structures:

8. The compound of claim 5 or a pharmaceutically acceptable salt thereof, wherein -L 2 -(R 7 ) n Groups with the following structures:

9. The compound of claim 5 or a pharmaceutically acceptable salt thereof, wherein L 2 For C6-C 10 Aryl.

10. The compound of claim 5 or a pharmaceutically acceptable salt thereof, wherein at least one R 7 The compounds are cyclopropyl, 2,2-difluorocyclopropyl, difluoromethoxy, 2,6-dimethylmorpholin-4-yl, N-azacyclobutyl, 3-fluorocyclobutyl, 2-methoxyethyl, ethoxy, methoxy, 2,2-difluoroethoxy, 2,2-difluoroethyl, trifluoromethyl, isopropyl, methyl, acetyl, fluorine, chlorine, 1-methylpyrazol-3-yl, dimethylamino, N-methyl-N-(2-methoxyethyl)-amino, N-ethyl-N-(2-methoxyethyl)-amino, N-(2-propyl)-N-(2-methoxyethyl)-amino, 2-methoxyethylamino, 3-aza-8-oxa-bicyclo[4.3]. [0] Non-3-yl, 3-aza-7-oxa-bicyclo[4.3.0] Non-3-yl, 1-fluorocyclobut-1-yl, 3-fluoropyrrolidine-1-yl, 3-methoxypyrrolidine-1-yl, oxacyclobutane-3-yl, N-methyldihydroindole-4-yl, 2,2-difluoro-3-methylcycloprop-1-yl, 3-methoxyazacyclobutane-1-yl, 3-methoxypiperidin-1-yl, 1,2-dimethyl-7-azaindole-4-yl, 1-methyl-7-azaindole-4-yl, 2,3-methylenedioxyphenyl, N-methyl-N-(3-oxacyclobutyl)amino, 3-oxacyclobutoxy, 1,1 -difluoro-5-azaspiro[2.3]hex-5-yl, 1-fluoromethyl-cyclopropyl, N-(3-tetrahydrofuranyl)methylamino, N-dihydroindolyl, N-1,4-oxazaheptanylcycloyl, 2-fluoro-2-propyl, 1,1-difluoro-2-propyl, 2,2-difluoro-1-methylcyclopropyl-1-yl, 1-methylcyclopropyl, 4,4-difluoropiperidin-1-yl, 2-methoxyethoxy, 3,3-difluorocyclobut-1-yl, N-methyl-N-1-methoxypropyl-2-ylamino, 1-methoxypropyl-2-ylamino, 1-methoxyethyl, 4-methylpiperazinyl, 3-methylmorpholinyl, 2,2-difluoropropyl Oxygen, 3-methoxycyclobutyl, methylamino, 4-dimethylamino-3,3-difluoropiperidinyl, 4-methylamino-3,3-difluoropiperidinyl, 3,3-difluoropyrrolidinyl, N-methyl-N-3-methoxycyclobutylamino, 1-methylpyrazol-5-yl, 6-oxa-3-azabicyclo[3.1.1]hept-3-yl, cyclopropoxy, 2,6-dimethylpyridin-4-yl, 2-methylpyrrolidinyl, 4-oxabicyclo[4.1.0]hept-1-yl, N-methyl-N-(2,6-dimethyltetrahydropyran-4-yl)amino or N-methyl-N-3-methyloxacyclobutane-3-ylmethylamino.

11. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R 1 It is hydrogen.

12. Compounds selected from the group consisting of compounds and their pharmaceutically acceptable salts.

13. Compounds selected from the group consisting of compounds and their pharmaceutically acceptable salts.

14. A pharmaceutical composition comprising a compound of any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

15. Use of the compound of any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating conditions related to the BAF complex.

16. The use of claim 15, wherein the condition associated with the BAF complex is cancer or a viral infection.

17. Use of a compound of any one of claims 1-13 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating cancer in a subject in need.

18. The use of claim 17, wherein the cancer is non-small cell lung cancer, colorectal cancer, bladder cancer, primary cancer of unknown cause, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, esophageal and gastric cancer, pancreatic cancer, hepatobiliary cancer, soft tissue sarcoma, ovarian cancer, head and neck cancer, renal cell carcinoma, bone cancer, non-Hodgkin's lymphoma, small cell lung cancer, prostate cancer, embryonal carcinoma, germ cell tumor, cervical cancer, thyroid cancer, salivary gland cancer, gastrointestinal neuroendocrine tumor, uterine sarcoma, gastrointestinal stromal tumor, CNS cancer, thymoma, adrenocortical carcinoma, appendix cancer, small bowel cancer, or penile cancer.

19. Use of a compound of any one of claims 1-13 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma or blood cancer in subjects in need.

20. Use of any compound of claims 1-13 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for reducing the levels and / or activity of BRG1 and / or BRM in melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma or hematologic malignancies.

21. The use of claim 17 or 20, further comprising administering the anticancer therapy to the subject or exposing the cells to the anticancer therapy.

22. The use of claim 21, wherein the anticancer therapy is a chemotherapeutic agent or cytotoxic agent, immunotherapy, surgery, radiotherapy, thermotherapy or photocoagulation or a combination thereof.

23. The use of claim 22, wherein one or more chemotherapeutic agents or cytotoxic agents are dacarbazine, temozolomide, cisplatin, treoxazine, formustin, IMCgp100, CTLA-4 inhibitors, PD-1 inhibitors, PD-L1 inhibitors, mitogen-activated protein kinase inhibitors and / or protein kinase C inhibitors.

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