Pyrazine derivatives and uses thereof

Abstract

The present disclosure features compounds of Formula I or II: # imgabs0 # or a pharmaceutically acceptable salt thereof, and formulations containing the same. Methods of treating BAF complex related disorders such as cancer are also disclosed.

Description

[0001] background

[0002] The present invention relates to compounds useful for modulating the BRG1- or BRM-associated factor (BAF) complex. In particular, the present invention relates to compounds useful for treating disorders associated with the function of the BAF complex.

[0003] Chromatin regulation is essential for gene expression, and ATP-dependent chromatin remodeling is a mechanism by which such gene expression occurs. The human switch / sucrose non-fermentable (SWI / SNF) chromatin remodeling complex (also known as the BAF complex) has two SWI2-like ATPases, called BRG1 (Brahma-related gene-1) and BRM (Brahma). The transcription 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 cancer tumors and is required for cancer cell proliferation. BRM (also known as the possible global transcription activator SNF2L2 and / or 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 BRG1 loss-of-function mutations. Inactivation of BRG and / or BRM leads to downstream effects in cells, including cell cycle arrest and tumor suppression.

[0004] Overview

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

[0006] In one aspect, the invention features a compound having a structure of Formula I or II, or a pharmaceutically acceptable salt thereof:

[0007]

[0008] in

[0009] Ring system A is a 5- to 9-membered heterocyclyl or heteroaryl;

[0010] m is 0, 1, 2, or 3;

[0011] k is 0, 1, or 2;

[0012] Each R 1 are independently halogen, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 Heteroalkyl, optionally substituted C 2 -C 9 Heterocyclic group, optionally substituted C 3 -C 8 Cycloalkyl or optionally substituted CH 2 -C 3 -C 8 Cycloalkyl;

[0013] each X is independently halogen;

[0014] L is a linker; and

[0015] B is the degradation part.

[0016] In some embodiments, the compound has the structure of Formula I or II:

[0017]

[0018] in

[0019] Ring system A is a 5- to 9-membered heterocyclyl or heteroaryl;

[0020] m is 0, 1, 2, or 3;

[0021] k is 0, 1, or 2;

[0022] L is the connector;

[0023] B is the degradation part;

[0024] Each R 1 are independently halogen, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 Heteroalkyl, optionally substituted C 3 -C 8 Cycloalkyl or optionally substituted C 2 -C 9 Heterocyclic group;

[0025] and

[0026] Each X is independently a halogen,

[0027] or a pharmaceutically acceptable salt thereof.

[0028] In some embodiments, the compound has the structure of Formula I or II:

[0029]

[0030] in

[0031] Ring system A is a 5- to 9-membered heterocyclyl or heteroaryl;

[0032] m is 0, 1, 2, or 3;

[0033] k is 0, 1, or 2;

[0034] L is a linker of formula III:

[0035] A 1 -(B 1 ) f -(C 1 ) g -(B 2 ) h -(B 3 ) i -(C 2 ) j -(B 4 ) k –A 2 ,

[0036] Formula III

[0037] in

[0038] A 1 is the bond between the linker and the ring system A;

[0039] A 2 is the bond between the degradable moiety and the linker;

[0040] B 1 , B 2 , B 3 and B 4 are each independently an optionally substituted ethynyl, an optionally substituted C 6 -C 10 Aryl, optionally substituted C 3 -C 10 Cycloalkyl, optionally substituted C 2 -C 9 Heterocyclic group, optionally substituted C 2 -C 9 Heteroaryl, O, S, S(O) 2 or NR N ;

[0041] Each R N are independently H, optionally substituted C 1–4 Alkyl, optionally substituted C 2–4 Alkenyl, optionally substituted C 2–4 Alkynyl, optionally substituted C2–6 Heterocyclic group, optionally substituted C 6–12 Aryl or optionally substituted C 1–7 heteroalkyl;

[0042] C 1 and C 2 are each independently carbonyl, thiocarbonyl, sulfonyl or phosphoryl; and

[0043] f, g, h, i, j and k are each independently 0 or 1.

[0044] B is the degradation part;

[0045] Each R 1 are independently halogen, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 Heteroalkyl, optionally substituted C 3 -C 8 Cycloalkyl or optionally substituted C 2 -C 9 Heterocyclic group;

[0046] and

[0047] Each X is independently a halogen,

[0048] or a pharmaceutically acceptable salt thereof.

[0049] In some embodiments, the compound has the structure of Formula IA or II-A:

[0050]

[0051] The dotted bonds represent single bonds or double bonds.

[0052] or a pharmaceutically acceptable salt thereof.

[0053] In some embodiments, the compound has the structure of Formula IG or II-G:

[0054]

[0055] or a pharmaceutically acceptable salt thereof.

[0056] In some embodiments, the compound has the structure of Formula IH or II-H:

[0057]

[0058] or a pharmaceutically acceptable salt thereof.

[0059] In some embodiments, m is 0 or 1.

[0060] In some embodiments, m is 1.

[0061] In some embodiments, R 1 is halogen, optionally substituted C 1 -C 6 Alkyl or optionally substituted C 3 -C 8 Cycloalkyl.

[0062] In some embodiments, R 1 It's methyl.

[0063] In some embodiments, R 1 It's cyclopropane.

[0064] In some embodiments, m is 0.

[0065] In some embodiments, k is 1.

[0066] In some embodiments, X is Cl.

[0067] In some embodiments, k is 0.

[0068] In some embodiments, the linker has the structure –(L 1 ) n -, where n is 1, 2 or 3, and each L 1 O, NR independently N , ethynyl, optionally substituted C 2 -C 10 Heterocyclic group, optionally substituted C 2 -C 9 Heteroaryl, optionally substituted C 6 -C 10 Aryl or optionally substituted C 3 -C 10 Cycloalkyl.

[0069] In some embodiments, at least one L 1 is an optionally substituted C 2 -C 10 In some embodiments, the optionally substituted C 2 -C 10 The heterocyclyl group is a 4-, 5- or 6-membered monocyclic heterocyclyl group. In some embodiments, the 4-, 5- or 6-membered monocyclic heterocyclyl group is:

[0070]

[0071]

[0072] In some embodiments, the optionally substituted C 2 -C10 The heterocyclic group is a spirocyclic heterocyclic group. In some embodiments, the spirocyclic heterocyclic group is:

[0073]

[0074] In some embodiments, the optionally substituted C 2 -C 10 The heterocyclyl group is a bridged heterocyclyl group. In some embodiments, the bridged heterocyclyl group is:

[0075]

[0076] In some embodiments, the optionally substituted C 2 -C 10 The heterocyclyl is a fused bicyclic heterocyclyl. In some embodiments, the fused bicyclic heterocyclyl is:

[0077]

[0078] In some embodiments, at least one L 1 is an optionally substituted C 2 -C 9 In some embodiments, the linker is -(L 1 ) q -(optionally substituted C 2 -C 9 Heteroaryl)-(L 1 ) q -, wherein each q is independently 0 or 1. In some embodiments, the optionally substituted C 2 -C 9 Heteroaryl is a 6-membered monocyclic heteroaryl. In some embodiments, the 6-membered monocyclic heteroaryl is:

[0079]

[0080] In some embodiments, at least one L 1 is an optionally substituted C 2 -C 9 In some embodiments, the linker is:

[0081]

[0082] In some embodiments, at least one L 1 is an optionally substituted C 6 -C 10 In some embodiments, the optionally substituted C 6 -C 10 Aryl is a 6-membered monocyclic aryl. In some embodiments, the 6-membered monocyclic aryl is an optionally substituted phenyl.

[0083] In some embodiments, at least one L 1 is an optionally substituted C 3 -C 10 In some embodiments, the optionally substituted C 3 -C 10 Cycloalkyl is a monocyclic cycloalkyl. In some embodiments, the 6-membered monocyclic cycloalkyl is:

[0084]

[0085] In some embodiments, the optionally substituted C 3 -C 10 Cycloalkyl is a bridged cycloalkyl. In some embodiments, the bridged cycloalkyl is:

[0086]

[0087] In some embodiments, at least one L 1 It is ethynyl.

[0088] In some embodiments, one and only one L 1 is O. In some embodiments, one and only one L 1 YesNR N In some embodiments, R N is an optionally substituted C 1 -C 4 In some embodiments, R N It's H.

[0089] In some embodiments, the linker has the following structure:

[0090] A 1 -(B 1 ) f -(B 2 ) h -(B 3 ) i -(B 4 ) k –A 2 ,

[0091] Among them B 1 , B 2 , B 3 and B 4 are each independently an optionally substituted ethynyl, an optionally substituted C 6 -C 10 Aryl, optionally substituted C 3 -C 10 Cycloalkyl, optionally substituted C 2-C 10 Heterocyclic group, optionally substituted C 2 -C 9 Heteroaryl, O or NR N .

[0092] In some embodiments, at least one of f, h, i, and k is 1.

[0093] In some embodiments, B 1 , B 2 , B 3 and B 4 are each independently O, ethynyl, optionally substituted C 2 -C 9 Heteroaryl, optionally substituted C 2 -C 10 Heterocyclic group, optionally substituted C 3 -C 10 Cycloalkyl or optionally substituted C 6 -C 10 In some embodiments, B 1 , B 2 , B 3 and B 4 are each independently an optionally substituted C 2 -C 9 Heteroaryl or optionally substituted C 2 -C 10 In some embodiments, B 1 and B 4 Each independently is

[0094]

[0095]

[0096] In some embodiments, B 1 yes:

[0097]

[0098]

[0099] In some embodiments, B 4 yes:

[0100]

[0101]

[0102] In some embodiments, B 2 YesNR N In some embodiments, B 2is NH. In some embodiments, B 2 is an optionally substituted C 2 -C 9 In some embodiments, B 2 yes:

[0103]

[0104] In some embodiments, f is 0. In some embodiments, f is 1. In some embodiments, g is 0. In some embodiments, g is 1. In some embodiments, h is 0. In some embodiments, h is 1. In some embodiments, i is 0. In some embodiments, i is 1. In some embodiments, j is 0. In some embodiments, j is 1. In some embodiments, k is 0. In some embodiments, k is 1.

[0105] In some embodiments, the linker has the structure

[0106]

[0107]

[0108]

[0109]

[0110] In one aspect, the invention features a compound having a structure of Formula I or II, or a pharmaceutically acceptable salt thereof:

[0111]

[0112] in

[0113] Ring system A is a 5- to 9-membered heterocyclyl or heteroaryl;

[0114] m is 0, 1, 2, or 3;

[0115] k is 0, 1, or 2;

[0116] R 1 is halogen, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 3 -C 8 Cycloalkyl or optionally substituted C 3 -C 10 Carbocyclic group;

[0117] X is a halogen;

[0118] L is a linker of formula IIIa:

[0119] A 1 -(B 1 ) f -(C 1 ) g -(B 2 ) h -(D)-(B 3 ) i -(C 2 ) j -(B 4 ) k –A 2 ,

[0120] Formula IIIa

[0121] or a pharmaceutically acceptable salt thereof,

[0122] in

[0123] A 1 is the bond between the linker and the ring system A;

[0124] A 2 is the bond between the degradable moiety and the linker;

[0125] B 1 , B 2 , B 3 and B 4 are each independently an optionally substituted C 1 -C 4 Alkyl, optionally substituted C 6 -C 10 Aryl, optionally substituted C 6 -C 10 Aryl C 1-4 Alkyl, optionally substituted C 1 -C 4 Heteroalkyl, optionally substituted C 3 -C 10 Cycloalkyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 2 -C 8 Heterocyclic group, optionally substituted C 2 -C 6 Heteroaryl, optionally substituted C 6–12 Aryl, O, S, S(O) 2 or NR N ;

[0126] Each R N are independently H, optionally substituted C 1–4 Alkyl, optionally substituted C 2–4 Alkenyl, optionally substituted C 2–4Alkynyl, optionally substituted C 2–10 Heterocyclic group, optionally substituted C 2–6 Heteroaryl or optionally substituted C 1–7 heteroalkyl;

[0127] C 1 and C 2 are each independently carbonyl, thiocarbonyl, sulfonyl or phosphoryl;

[0128] f, g, h, i, j and k are each independently 0 or 1; and

[0129] D is optionally substituted C 1–10 Alkyl, optionally substituted C 2–10 Alkenyl, optionally substituted C 2–10 Alkynyl, optionally substituted C 2–10 Heterocyclic group, optionally substituted C 2–6 Heteroaryl, optionally substituted C 6–12 Aryl, optionally substituted C 2 -C 10 Polyethylene glycol, optionally substituted C 3 -C 10 Cycloalkyl, optionally substituted C 3 -C 10 Carbocyclic or optionally substituted C 1–10 heteroalkyl; or D is absent and the linker is A 1 -(B 1 ) f -(C 1 ) g -(B 2 ) h -(B 3 ) i -(C 2 ) j -(B 4 ) k –A 2 .

[0130] In some embodiments, B 1 , B 2 , B 3 and B 4 are each independently an optionally substituted C 1 -C 2 Alkyl, optionally substituted C 1 -C 3 Heteroalkyl, optionally substituted C 2 -C 10 Heterocyclic group, optionally substituted C 2–6 Heteroaryl, O or NR N ; and D is optionally substituted C 1–10Alkyl, optionally substituted C 2–10 Alkenyl, optionally substituted C 2–10 Alkynyl, optionally substituted C 2–10 Heterocyclic group, optionally substituted C 6–12 Aryl, optionally substituted C 2 -C 10 Polyethylene glycol or optionally substituted C 1–10 Heteroalkyl or A 1 -(B 1 ) f -(C 1 ) g -(B 2 ) h -Connect to-(B 3 ) i -(C 2 ) j -(B 4 ) k –A 2 chemical bonds.

[0131] In some embodiments, B 1 , B 2 , B 3 and B 4 are each independently an optionally substituted C 1 -C 2 Alkyl, optionally substituted C 1 -C 3 Heteroalkyl, optionally substituted C 2 -C 10 Heterocyclic group, optionally substituted C 2–6 Heteroaryl, optionally substituted C 3 -C 10 Cycloalkyl, optionally substituted C 3 -C 10 Carbocyclic group, O or NR N .

[0132] In some embodiments, B 1 and B 4 Each independently is

[0133]

[0134]

[0135]

[0136] In some embodiments, B 1 yes

[0137]

[0138]

[0139]

[0140]

[0141] In some embodiments, B 4 yes

[0142]

[0143]

[0144]

[0145] In some embodiments, C 1 yes

[0146] In some embodiments, B 2 is an optionally substituted C 1 -C 4 alkyl.

[0147] In some embodiments, D is optionally substituted C 1 -C 10 alkyl.

[0148] In some embodiments, f is 1. In some embodiments, g is 0. In some embodiments, g is 1. In some embodiments, h is 0. In some embodiments, h is 1. In some embodiments, i is 0. In some embodiments, i is 1. In some embodiments, j is 0. In some embodiments, j is 1. In some embodiments, k is 0. In some embodiments, k is 1.

[0149] In some embodiments, D is absent and the linker is A 1 -(B 1 ) f -(C 1 ) g -(B 2 ) h -(B 3 ) i -(C 2 ) j -(B 4 ) k –A 2 .

[0150] In some embodiments, the linker is D. In some embodiments, D is an optionally substituted C 1–10 Alkyl, optionally substituted C2–10 Alkenyl, optionally substituted C 2–10 Alkynyl, optionally substituted C 2–10 Heterocyclic group, optionally substituted C 2–6 Heteroaryl, optionally substituted C 6–12 Aryl, optionally substituted C 2 -C 10 Polyethylene glycol or optionally substituted C 1–10 In some embodiments, D is an optionally substituted C 3 -C 10 cycloalkyl, f is 1, g is 0, h is 0, i is 0, j is 0, and k is 1. In some embodiments, D is an optionally substituted C 3 -C 10 cycloalkyl, f is 1, g is 0, h is 0, i is 0, j is 0, and k is 0. In some embodiments, D is an optionally substituted C 3 -C 10 cycloalkyl, f is 0, g is 0, h is 0, i is 0, j is 0, and k is 1. In some embodiments, D is an optionally substituted C 3 -C 10 cycloalkyl, f is 0, g is 0, h is 0, i is 0, j is 0, and k is 0. In some embodiments, D is an optionally substituted C 3 -C 10 carbocyclyl, f is 1, g is 0, h is 0, i is 0, j is 0, and k is 1. In some embodiments, D is an optionally substituted C 3 -C 10 carbocyclyl, f is 1, g is 0, h is 0, i is 0, j is 0, and k is 0. In some embodiments, D is an optionally substituted C 3 -C 10 carbocyclyl, f is 0, g is 0, h is 0, i is 0, j is 0, and k is 1. In some embodiments, D is an optionally substituted C 3 -C 10 carbocyclyl, f is 0, g is 0, h is 0, i is 0, j is 0, and k is 0. In some embodiments, D is:

[0151]

[0152]

[0153]

[0154] In some embodiments, the linker has the structure

[0155]

[0156]

[0157]

[0158] In some embodiments, the compound has the structure of Formula IA or II-A:

[0159]

[0160] The dashed bonds represent single bonds or double bonds.

[0161] In some embodiments, the compound has the structure of Formula IG or II-G:

[0162]

[0163] Where m is 0 or 1;

[0164] R 1 is halogen, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 3 -C 8 Cycloalkyl or optionally substituted C 3 -C 10 Carbocyclic group;

[0165] k is 0 or 1; and

[0166] X is halogen.

[0167] In some embodiments, the compound has the structure of Formula IH or II-H:

[0168]

[0169] in

[0170] m is 0 or 1;

[0171] R 1 is halogen, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 3 -C 8 Cycloalkyl or optionally substituted C 3 -C 10 Carbocyclic group.

[0172] k is 0 or 1; and

[0173] X is halogen.

[0174] In some embodiments, m is 0.

[0175] In some embodiments, m is 1.

[0176] In some embodiments, R 1 is an optionally substituted C 1 -C 6 In some embodiments, R 1 It's methyl.

[0177] In some embodiments, R 1 is an optionally substituted C 3 -C 8 In some embodiments, R 1 is an optionally substituted C 3 -C 10 In some embodiments, R 1 It's cyclopropane.

[0178] In some embodiments, k is 0.

[0179] In some embodiments, k is 1. In some embodiments, X is Cl.

[0180] In some embodiments, the degradation moiety B has the structure of Formula A-1:

[0181]

[0182] in

[0183] Y 1 yes

[0184] R A5 is H, optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0185] R A6 is H or optionally substituted C 1 -C 6 alkyl; and R A7 is H or optionally substituted C 1 -C 6 Alkyl; or R A6 and R A7 are combined together with the carbon atoms to which they are each bonded to form an optionally substituted C 3 -C 6 Carbocyclic or optionally substituted C 2 -C 5 heterocyclic group; or R A6 and R A7 are combined together with the carbon atoms to which they are each bonded to form an optionally substituted C 3 -C 6Carbocyclic or optionally substituted C 2 -C 5 Heterocyclic group;

[0186] R A8 is H, optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0187] R A1 , R A2 , R A3 and R A4 Each independently is H, A 2 , halogen, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 Heteroalkyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 2 -C 9 Heterocyclic group, optionally substituted C 6 -C 10 Aryl, optionally substituted C 2 -C 9 Heteroaryl, optionally substituted C 2 -C 6 Alkenyl, optionally substituted C 2 -C 6 Heteroalkenyl, optionally substituted -OC 3 -C 6 carbocyclyl, hydroxyl, thiol or optionally substituted amino; or R A1 and R A2 , R A2 and R A3 , and / or R A3 and R A4 Combined with the carbon atoms to which they are attached to form and is an optionally substituted C 6 -C 10 Aryl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 2 -C 9 Heteroaryl or C 2 -C 9 Heterocyclic group, any one of which is optionally replaced by A 2 replace,

[0188] Where R A1 , R A2 , RA3 and R A4 One of them is A 2 ,or A 2 Replacement; and

[0189] A 2 is the bond between the degradable moiety and the linker.

[0190] In some embodiments, R A5 is H or methyl. In some embodiments, R A5 It's H.

[0191] In some embodiments, R A1 , R A2 , R A3 and R A4 Each independently is H or A 2 .

[0192] In some embodiments, R A1 Yes A 2 And R A2 , R A3 and R A4 Each is H.

[0193] In some embodiments, R A2 Yes A 2 And R A1 , R A3 and R A4 Each is H.

[0194] In some embodiments, R A3 Yes A 2 And R A1 , R A2 and R A4 Each is H.

[0195] In some embodiments, R A4 Yes A 2 And R A1 , R A2 and R A3 Each is H.

[0196] In some embodiments, Y 1 yes

[0197] In some embodiments, R A6 is H. In some embodiments, R A7 It's H.

[0198] In some embodiments, Y 1 yes

[0199] In some embodiments, R A8 is H or optionally substituted C 1 -C 6 In some embodiments, R A8 is H or methyl. In some embodiments, R A8 It's methyl.

[0200] In some embodiments, the degradation moiety comprises the structure of Formula A2:

[0201]

[0202] In some embodiments, the degradation moiety is

[0203]

[0204] In some embodiments, the degradation moiety comprises the structure of Formula A4:

[0205]

[0206] In some embodiments, the degradation moiety is

[0207]

[0208] In some embodiments, the degradation moiety has the structure of Formula A5:

[0209]

[0210] In some embodiments, the degradation moiety has the structure of Formula A6:

[0211]

[0212] In some embodiments, the degradation moiety has the structure of Formula A8:

[0213]

[0214] In some embodiments, the degradation moiety has the structure of Formula A10:

[0215]

[0216] In some embodiments, the degradation moiety has the structure

[0217]

[0218] In some embodiments, the degradation moiety has the structure

[0219]

[0220] In some embodiments, the degradation moiety has the structure of Formula C:

[0221]

[0222] in

[0223] L 4 Yes-N(R B1 )(R B2 ),

[0224] R B1 It is H, A 2 , optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0225] R B2 is H, optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0226] R B3 Yes A 2 , optionally substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 Heteroalkyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 6 -C 10 Aryl, optionally substituted C 1 -C 6 Alkyl C 3 -C 10 Carbocyclic or optionally substituted C 1 -C 6 Alkyl C 6 -C 10 Aryl;

[0227] R B4 is H, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 6 -C 10 Aryl, optionally substituted C 1 -C 6 Alkyl C 3-C 10 Carbocyclic or optionally substituted C 1 -C 6 Alkyl C 6 -C 10 Aryl;

[0228] R B5 is H, optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0229] v2 is 0, 1, 2, 3, or 4;

[0230] Each R B6 Independently is A 2 , halogen, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 2 -C 6 Alkynyl, optionally substituted C 1 -C 6 Heteroalkyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 2 -C 9 Heterocyclic group, optionally substituted C 6 -C 10 Aryl, optionally substituted C 2 -C 9 Heteroaryl, optionally substituted C 2 -C 6 Alkenyl, optionally substituted C 2 -C 6 heteroalkenyl, hydroxy, thiol, cyano or optionally substituted amino;

[0231] R B7 and R B8 are each independently H, halogen, optionally substituted C 1 -C 6 Alkyl or optionally substituted C 6 -C 10 Aryl;

[0232] R B9 is H or optionally substituted C 1 -C 6 Alkyl; and

[0233] A 2 is the bond between the degradable moiety and the linker;

[0234] Where R B1 , R B3 and RB6 One and only one of them is A 2 ,

[0235] or a pharmaceutically acceptable salt thereof.

[0236] In some embodiments, the degradation moiety has the structure of Formula C:

[0237]

[0238] in

[0239] L 4 Yes-N(R B1 )(R B2 ),

[0240] R B1 It is H, A 2 , optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0241] R B2 is H, optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0242] R B3 Yes A 2 , optionally substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 Heteroalkyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 6 -C 10 Aryl, optionally substituted C 1 -C 6 Alkyl C 3 -C 10 Carbocyclic or optionally substituted C 1 -C 6 Alkyl C 6 -C 10 Aryl;

[0243] R B4 is H, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 6-C 10 Aryl, optionally substituted C 1 -C 6 Alkyl C 3 -C 10 Carbocyclic or optionally substituted C 1 -C 6 Alkyl C 6 -C 10 Aryl;

[0244] R B5 is H, optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0245] v2 is 0, 1, 2, 3, or 4;

[0246] Each R B6 Independently is A 2 , halogen, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 Heteroalkyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 2 -C 9 Heterocyclic group, optionally substituted C 6 -C 10 Aryl, optionally substituted C 2 -C 9 Heteroaryl, optionally substituted C 2 -C 6 Alkenyl, optionally substituted C 2 -C 6 heteroalkenyl, hydroxy, thiol or optionally substituted amino;

[0247] R B7 and R B8 are each independently H, halogen, optionally substituted C 1 -C 6 Alkyl or optionally substituted C 6 -C 10 Aryl;

[0248] R B9 is H or optionally substituted C 1 -C 6 alkyl;

[0249] R B10 is H or F; and

[0250] A 2is the bond between the degradable moiety and the linker;

[0251] Where R B1 , R B3 and R B6 One and only one of them is A 2 ,

[0252] or a pharmaceutically acceptable salt thereof.

[0253] In some embodiments, the degradation moiety has the structure of Formula C3:

[0254]

[0255] In some embodiments, the degradation moiety has the structure of Formula C4:

[0256]

[0257] In some embodiments, the degradation moiety has the structure of Formula C1:

[0258]

[0259] In some embodiments, the degradation moiety is

[0260]

[0261] In some embodiments, the degradation moiety is

[0262]

[0263] In some embodiments, the degradation moiety is

[0264]

[0265] In some embodiments, the degradation moiety is

[0266]

[0267] In some embodiments, the degradation moiety is

[0268]

[0269] In some embodiments, the degradation moiety is

[0270]

[0271] In some embodiments, the degradation moiety has the structure of Formula C2:

[0272]

[0273] In some embodiments, RB9 is an optionally substituted C 1 -C 6 In some embodiments, R B9 It's methyl.

[0274] In some embodiments, R B9 Bonded to the (S)-stereocenter.

[0275] In some embodiments, v2 is 0. In some embodiments, R B4 is H. In some embodiments, R B5 is H. In some embodiments, R B7 is an optionally substituted C 1 -C 6 In some embodiments, R B7 In some embodiments, R B3 is an optionally substituted C 1 -C 6 In some embodiments, R B3 In some embodiments, R B8 is H. In some embodiments, R B2 It's H.

[0276] In some embodiments, the degradation moiety is

[0277]

[0278] In some embodiments, the degradation moiety has the structure of formula Ca2:

[0279]

[0280] In some embodiments, the degradation moiety has the structure of Formula Cb2:

[0281]

[0282] In some embodiments, the degradation moiety has the structure of Formula Cc2:

[0283]

[0284] In some embodiments, the degradation moiety has the structure of formula Cd2:

[0285]

[0286] In some embodiments, the degradation moiety has the structure of formula Ce2:

[0287]

[0288] In some embodiments, the degradation moiety has the structure of Formula Cf2:

[0289]

[0290] In some embodiments, R B9 is an optionally substituted C 1 -C 6 In some embodiments, R B9 It's methyl.

[0291] In some embodiments, R B9 Bonded to the (S)-stereocenter.

[0292] In some embodiments, v2 is 0. In some embodiments, R B4 is H. In some embodiments, R B5 is H. In some embodiments, R B7 is an optionally substituted C 1 -C 6 In some embodiments, R B7 In some embodiments, R B3 is an optionally substituted C 1 -C 6 In some embodiments, R B3 In some embodiments, R B3 is an optionally substituted C 3 -C 10 In some embodiments, R B3 In some embodiments, R B3 In some embodiments, R B3 is fluoro-2-methylpropane. B8 is H. In some embodiments, R B2 It's H.

[0293] In some embodiments, the degradation moiety is

[0294]

[0295] In some embodiments, the degradation moiety is

[0296]

[0297] In some embodiments, the degradation moiety is

[0298]

[0299] In some embodiments, the degradation moiety is

[0300]

[0301] In some embodiments, the degradation moiety is

[0302]

[0303] In some embodiments, the degradation moiety is

[0304]

[0305] In some embodiments, the degradation moiety is

[0306]

[0307] In some embodiments, the degradation moiety is

[0308]

[0309] In some embodiments, the degradation moiety is

[0310]

[0311] In some embodiments, the degradation moiety has the structure of Formula C5:

[0312]

[0313] in

[0314] L 4 Yes-N(R B1 )(R B2 ),

[0315] R B1 It is H, A 2 , optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0316] R B2 is H, optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0317] R B3 Yes A 2 , optionally substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C6 Heteroalkyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 6 -C 10 Aryl, optionally substituted C 1 -C 6 Alkyl C 3 -C 10 Carbocyclic or optionally substituted C 1 -C 6 Alkyl C 6 -C 10 Aryl;

[0318] R B5 is H, optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0319] v2 is 0, 1, 2, 3, or 4;

[0320] Each R B6 Independently is A 2 , halogen, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 2 -C 6 Alkynyl, optionally substituted C 1 -C 6 Heteroalkyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 2 -C 9 Heterocyclic group, optionally substituted C 6 -C 10 Aryl, optionally substituted C 2 -C 9 Heteroaryl, optionally substituted C 2 -C 6 Alkenyl, optionally substituted C 2 -C 6 heteroalkenyl, hydroxy, thiol, cyano or optionally substituted amino;

[0321] R B7 and R B8 are each independently H, halogen, optionally substituted C 1 -C 6 Alkyl or optionally substituted C 6 -C 10 Aryl;

[0322] R B9 is H or optionally substituted C1 -C 6 alkyl;

[0323] R B11 is H, alcohol, boronic acid, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 6 -C 10 Aryl, optionally substituted C 1 -C 6 Alkyl C 3 -C 10 Carbocyclic or optionally substituted C 1 -C 6 Alkyl C 6 -C 10 Aryl;

[0324] and

[0325] A 2 is the bond between the degradable moiety and the linker;

[0326] Where R B1 , R B3 and R B6 One and only one of them is A 2 ,

[0327] or a pharmaceutically acceptable salt thereof.

[0328] In some embodiments, R B11 It's boric acid.

[0329] In some embodiments, the degradation moiety has the structure of Formula C6:

[0330]

[0331] In some embodiments, the degradation moiety has the structure of Formula C1:

[0332]

[0333] In some embodiments, the degradation moiety has the structure of Formula C8:

[0334]

[0335] In some embodiments, R B9 is an optionally substituted C 1 -C 6 In some embodiments, R B9 It's methyl.

[0336] In some embodiments, R B9 Bonded to the (S)-stereocenter.

[0337] In some embodiments, v2 is 0. In some embodiments, R B5 is H. In some embodiments, R B7 is an optionally substituted C 1 -C 6 In some embodiments, R B7 In some embodiments, R B3 is an optionally substituted C 1 -C 6 In some embodiments, R B3 In some embodiments, R B8 is H. In some embodiments, R B2 It's H.

[0338] In some embodiments, the degradation moiety is

[0339]

[0340] In some embodiments, the degradation moiety has the structure of Formula D:

[0341]

[0342] in

[0343] L 4 Yes-N(R B1 )(R B2 ),

[0344] R B1 It is H, A 2 , optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0345] R B2 is H, optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0346] R B3 Yes A 2 , optionally substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C6 Heteroalkyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 6 -C 10 Aryl, optionally substituted C 1 -C 6 Alkyl C 3 -C 10 Carbocyclic or optionally substituted C 1 -C 6 Alkyl C 6 -C 10 Aryl;

[0347] R B4 is H, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 6 -C 10 Aryl, optionally substituted C 1 -C 6 Alkyl C 3 -C 10 Carbocyclic or optionally substituted C 1 -C 6 Alkyl C 6 -C 10 Aryl;

[0348] R B5 is H, optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0349] v2 is 0, 1, 2, 3, or 4;

[0350] Each R B6 Independently is A 2 , halogen, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 Heteroalkyl, optionally substituted C 2 -C 6 Alkynyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 2 -C 9 Heterocyclic group, optionally substituted C 6 -C 10 Aryl, optionally substituted C 2 -C 9Heteroaryl, optionally substituted C 2 -C 6 Alkenyl, optionally substituted C 2 -C 6 heteroalkenyl, hydroxy, thiol, cyano or optionally substituted amino;

[0351] R B9 is H or optionally substituted C 1 -C 6 Alkyl; and

[0352] A 2 is the bond between the degradable moiety and the linker;

[0353] Where R B1 , R B3 and R B6 One and only one of them is A 2 ,

[0354] or a pharmaceutically acceptable salt thereof.

[0355] In some embodiments, the degradation moiety has the structure of formula D3.

[0356]

[0357] In some embodiments, the degradation moiety has the structure of Formula D1:

[0358]

[0359] In some embodiments, the degradation moiety is

[0360]

[0361] In some embodiments, the degradation moiety is

[0362]

[0363] In some embodiments, the degradation moiety is

[0364]

[0365] In some embodiments, the degradation moiety has the structure of Formula D2:

[0366]

[0367] In some embodiments, R B9 is an optionally substituted C 1 -C 6 In some embodiments, R B9 It's methyl.

[0368] In some embodiments, R B9 Bonded to the (S)-stereocenter. In some embodiments, R B9 It's H.

[0369] In some embodiments, v2 is 0. In some embodiments, v2 is 1. In some embodiments, v2 is 2. In some embodiments, R B4 is H. In some embodiments, R B5 is H. In some embodiments, R B3 is an optionally substituted C 1 -C 6 In some embodiments, R B3 In some embodiments, R B6 is H. In some embodiments, R B6 In some embodiments, R B6 In some embodiments, R B6 In some embodiments, R B6 In some embodiments, R B6 In some embodiments, R B6 is an optionally substituted C 1 -C 6 In some embodiments, R B6 is an optionally substituted C 3 -C 6 In some embodiments, R B6 In some embodiments, R B6 It is 3-methoxy-1-propanoxy.

[0370] In some embodiments, the degradation moiety is

[0371]

[0372] In some embodiments, the degradation moiety is

[0373]

[0374] In some embodiments, the degradation moiety is

[0375]

[0376] In some embodiments, the degradation moiety is

[0377]

[0378] In some embodiments, the degradation moiety is

[0379]

[0380] In some embodiments, the degradation moiety is

[0381]

[0382] In some embodiments, the degradation moiety is

[0383]

[0384] In some embodiments, the degradation moiety is

[0385]

[0386] In some embodiments, the degradation moiety is

[0387]

[0388] In some embodiments, the degradation moiety is

[0389]

[0390] In some embodiments, the degradation moiety is

[0391]

[0392] In some embodiments, the degradation moiety is

[0393]

[0394] In some embodiments, the degradation moiety is

[0395]

[0396] In some embodiments, the degradation moiety is

[0397]

[0398] In some embodiments, the degradation moiety is

[0399]

[0400] In some embodiments, the degradation moiety is

[0401]

[0402] In some embodiments, the degradation moiety is

[0403]

[0404] In some embodiments, the degradation moiety is

[0405]

[0406] In some embodiments, the degradation moiety is

[0407]

[0408] In some embodiments, the degradation moiety is

[0409]

[0410] In some embodiments, the degradation moiety is

[0411]

[0412] In some embodiments, the degradation moiety is

[0413]

[0414] In some embodiments, the degradation moiety is

[0415]

[0416] In some embodiments, the degradation moiety is

[0417]

[0418] In some embodiments, the degradation moiety is

[0419]

[0420] In some embodiments, the degradation moiety has the structure of Formula Da:

[0421]

[0422] in

[0423] L 4 Yes-N(R B1 )(R B2 ),

[0424] R B1 It is H, A 2 , optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0425] R B2is H, optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0426] R B3 Yes A 2 , optionally substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 Heteroalkyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 6 -C 10 Aryl, optionally substituted C 1 -C 6 Alkyl C 3 -C 10 Carbocyclic or optionally substituted C 1 -C 6 Alkyl C 6 -C 10 Aryl;

[0427] R B4 is H, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 6 -C 10 Aryl, optionally substituted C 1 -C 6 Alkyl C 3 -C 10 Carbocyclic or optionally substituted C 1 -C 6 Alkyl C 6 -C 10 Aryl;

[0428] R B5 is H, optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0429] X 1 and X 2 are each independently C, N or O;

[0430] v2 is 0, 1, 2, 3, or 4;

[0431] Each R B6 Each independently is A 2, halogen, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 2 -C 6 Alkynyl, optionally substituted C 1 -C 6 Heteroalkyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 2 -C 9 Heterocyclic group, optionally substituted C 6 -C 10 Aryl, optionally substituted C 2 -C 9 Heteroaryl, optionally substituted C 2 -C 6 Alkenyl, optionally substituted C 2 -C 6 heteroalkenyl, hydroxy, thiol, cyano or optionally substituted amino;

[0432] R B9 is H or optionally substituted C 1 -C 6 Alkyl; and

[0433] A 2 is the bond between the degradable moiety and the linker;

[0434] Where R B1 , R B3 and R B6 One and only one of them is A 2 ,

[0435] or a pharmaceutically acceptable salt thereof.

[0436] In some embodiments, the degradation moiety has the structure of formula Da3:

[0437]

[0438] In some embodiments, the degradation moiety has the structure of Formula Da1:

[0439]

[0440] In some embodiments, the degradation moiety has the structure of formula Da2:

[0441]

[0442] In some embodiments, R B9 is an optionally substituted C 1 -C 6 In some embodiments, R B9 It's methyl.

[0443] In some embodiments, R B9 Bonded to the (S)-stereocenter.

[0444] In some embodiments, v2 is 0. In some embodiments, R B4 is H. In some embodiments, R B5 is H. In some embodiments, R B3 is an optionally substituted C 1 -C 6 In some embodiments, R B3 In some embodiments, R B2 is H. In some embodiments, X 1 is C. In some embodiments, X 2 It is N.

[0445] In some embodiments, the degradation moiety is

[0446]

[0447] In some embodiments, the degradation moiety has the structure of Formula E:

[0448]

[0449] in

[0450] L 4 Yes-N(R B1 )(R B2 ),

[0451] R B1 It is H, A 2 , optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0452] R B2 is H, optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0453] R B3 Yes A 2 , optionally substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 Heteroalkyl, optionally substituted C3 -C 10 Carbocyclic group, optionally substituted C 6 -C 10 Aryl, optionally substituted C 1 -C 6 Alkyl C 3 -C 10 Carbocyclic or optionally substituted C 1 -C 6 Alkyl C 6 -C 10 Aryl;

[0454] R B4 is H, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 6 -C 10 Aryl, optionally substituted C 1 -C 6 Alkyl C 3 -C 10 Carbocyclic or optionally substituted C 1 -C 6 Alkyl C 6 -C 10 Aryl;

[0455] R B5 is H, optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0456] R B9 is H, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 3 -C 6 Alkynyl, optionally substituted C 3 -C 10 Carbocyclic or optionally substituted C 2 -C 10 Heterocyclic group;

[0457] B 10 is H, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 3 -C 6 Alkynyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 2 -C 10heterocyclic group, optionally substituted amino group or cyano group, and

[0458] A 2 is the bond between the degradable moiety and the linker;

[0459] Where R B1 , R B3 and R B6 One and only one of them is A 2 ,

[0460] or a pharmaceutically acceptable salt thereof.

[0461] In some embodiments, the degradation moiety has the structure of Formula E3:

[0462]

[0463] In some embodiments, the degradation moiety has the structure of Formula E1:

[0464]

[0465] In some embodiments, the degradation moiety is

[0466]

[0467] In some embodiments, the degradation moiety is

[0468]

[0469] In some embodiments, the degradation moiety has the structure of Formula E2:

[0470]

[0471] In some embodiments, R B9 is an optionally substituted C 1 -C 6 In some embodiments, R B9 It's methyl.

[0472] In some embodiments, R B9 Bonded to the (S)-stereocenter.

[0473] In some embodiments, v2 is 0. In some embodiments, v2 is 1. In some embodiments, R B4 is H. In some embodiments, R B5 is H. In some embodiments, R B3 is an optionally substituted C 1 -C 6 In some embodiments, R B3 In some embodiments, RB2 is H. In some embodiments, R B9 is an optionally substituted C 1 -C 6 In some embodiments, R B9 In some embodiments, R B9 is H. In some embodiments, R B9 is an optionally substituted C 3 -C 6 In some embodiments, R B10 In some embodiments, R B9 is [1.1.1]pentane. In some embodiments, R B9 In some embodiments, R B9 In some embodiments, R B9 In some embodiments, R B10 is H. In some embodiments, R B10 In some embodiments, R B10 is an optionally substituted C 3 -C 10 In some embodiments, R B10 is an optionally substituted C 1 -C 6 In some embodiments, R B10 It's methyl.

[0474] In some embodiments, the degradation moiety is

[0475]

[0476] In some embodiments, the degradation moiety is

[0477]

[0478] In some embodiments, the degradation moiety is

[0479]

[0480] In some embodiments, the degradation moiety is

[0481]

[0482] In some embodiments, the degradation moiety is

[0483]

[0484] In some embodiments, the degradation moiety is

[0485]

[0486] In some embodiments, the degradation moiety is

[0487]

[0488] In some embodiments, the degradation moiety is

[0489]

[0490] In some embodiments, the degradation moiety is

[0491]

[0492] In some embodiments, the degradation moiety has the structure of Formula F:

[0493]

[0494] in

[0495] L 4 Yes-N(R B1 )(R B2 ),

[0496] R B1 It is H, A 2 , optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0497] R B2 is H, optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0498] R B3 Yes A 2 , optionally substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 Heteroalkyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 6 -C 10 Aryl, optionally substituted C 1 -C 6 Alkyl C 3 -C 10 Carbocyclic or optionally substituted C1 -C 6 Alkyl C 6 -C 10 Aryl;

[0499] R B4 is H, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 6 -C 10 Aryl, optionally substituted C 1 -C 6 Alkyl C 3 -C 10 Carbocyclic or optionally substituted C 1 -C 6 Alkyl C 6 -C 10 Aryl;

[0500] R B5 is H, optionally substituted C 1 -C 6 Alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0501] A 2 is the bond between the degradable moiety and the linker;

[0502] Where R B1 or R B3 One and only one of them is A 2 ,

[0503] or a pharmaceutically acceptable salt thereof.

[0504] In some embodiments, the degradation moiety has the structure of Formula F3:

[0505]

[0506] In some embodiments, the degradation moiety has the structure of Formula F1:

[0507]

[0508] In some embodiments, the degradation moiety is

[0509]

[0510] In some embodiments, the degradation moiety is

[0511]

[0512] In some embodiments, the degradation moiety is

[0513]

[0514] In some embodiments, the degradation moiety has the structure of Formula F2:

[0515]

[0516] In some embodiments, R B9 is an optionally substituted C 1 -C 6 In some embodiments, R B9 It's methyl.

[0517] In some embodiments, R B4 is H. In some embodiments, R B5 is H. In some embodiments, R B3 is an optionally substituted C 1 -C 6 In some embodiments, R B3 In some embodiments, R B2 It's H.

[0518] In some embodiments, the degradation moiety is

[0519]

[0520] In some embodiments, the linker has the structure of Formula II:

[0521] A 1 -(B 1 ) f -(C 1 ) g -(B 2 ) h -(D)-(B 3 ) i -(C 2 ) j -(B 4 ) k –A 2 ,

[0522] Formula II

[0523] or a pharmaceutically acceptable salt thereof,

[0524] in

[0525] A 1 is the bond between the linker and the ring system A;

[0526] A 2is the bond between the degradable moiety and the linker;

[0527] B 1 , B 2 , B 3 and B 4 are each independently an optionally substituted C 1 -C 4 Alkyl, optionally substituted C 6 -C 10 Aryl, optionally substituted C 6 -C 10 Aryl C 1-4 Alkyl, optionally substituted C 1 -C 4 Heteroalkyl, optionally substituted C 3 -C 10 Cycloalkyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 2 -C 8 Heterocyclic group, optionally substituted C 2 -C 6 Heteroaryl, optionally substituted C 6–12 Aryl, O, S, S(O) 2 or NR N ;

[0528] Each R N are independently H, optionally substituted C 1–4 Alkyl, optionally substituted C 2–4 Alkenyl, optionally substituted C 2–4 Alkynyl, optionally substituted C 2–10 Heterocyclic group, optionally substituted C 2–6 Heteroaryl or optionally substituted C 1–7 heteroalkyl;

[0529] C 1 and C 2 are each independently carbonyl, thiocarbonyl, sulfonyl or phosphoryl;

[0530] f, g, h, i, j and k are each independently 0 or 1; and

[0531] D is optionally substituted C 1–10 Alkyl, optionally substituted C 2–10 Alkenyl, optionally substituted C 2–10 Alkynyl, optionally substituted C 2–10 Heterocyclic group, optionally substituted C 2–6 Heteroaryl, optionally substituted C 6–12 Aryl, optionally substituted C 2 -C 10 Polyethylene glycol, optionally substituted C3 -C 10 Cycloalkyl, optionally substituted C 3 -C 10 Carbocyclic or optionally substituted C 1–10 heteroalkyl; or D is absent and the linker is A 1 -(B 1 ) f -(C 1 ) g -(B 2 ) h -(B 3 ) i -(C 2 ) j -(B 4 ) k –A 2 .

[0532] In some embodiments, B 1 , B 2 , B 3 and B 4 are each independently an optionally substituted C 1 -C 2 Alkyl, optionally substituted C 1 -C 3 Heteroalkyl, optionally substituted C 2 -C 10 Heterocyclic group, optionally substituted C 2–6 Heteroaryl, O or NR N ; and D is optionally substituted C 1–10 Alkyl, optionally substituted C 2–10 Alkenyl, optionally substituted C 2–10 Alkynyl, optionally substituted C 2–10 Heterocyclic group, optionally substituted C 6–12 Aryl, optionally substituted C 2 -C 10 Polyethylene glycol or optionally substituted C 1–10 Heteroalkyl or A 1 -(B 1 ) f -(C 1 ) g -(B 2 ) h -Connect to-(B 3 ) i -(C 2 ) j -(B 4 ) k –A 2 chemical bonds.

[0533] In some embodiments, B1 , B 2 , B 3 and B 4 are each independently an optionally substituted C 1 -C 2 Alkyl, optionally substituted C 1 -C 3 Heteroalkyl, optionally substituted C 2 -C 10 Heterocyclic group, optionally substituted C 2–6 Heteroaryl, optionally substituted C 3 -C 10 Cycloalkyl, optionally substituted C 3 -C 10 Carbocyclic group, O or NR N .

[0534] In some embodiments, B 1 and B 4 Each independently is

[0535]

[0536]

[0537]

[0538] In some embodiments, B 1 yes

[0539]

[0540]

[0541]

[0542] In some embodiments, B 4 yes

[0543]

[0544]

[0545] In some embodiments, C 1 yes

[0546] In some embodiments, B 2 is an optionally substituted C 1 -C 4 alkyl.

[0547] In some embodiments, D is optionally substituted C 1 -C 10 alkyl.

[0548] In some embodiments, f is 1. In some embodiments, g is 0. In some embodiments, g is 1. In some embodiments, h is 0. In some embodiments, h is 1. In some embodiments, i is 0. In some embodiments, i is 1. In some embodiments, j is 0. In some embodiments, j is 1. In some embodiments, k is 0. In some embodiments, k is 1.

[0549] In some embodiments, D is absent and the linker is A 1 -(B 1 ) f -(C 1 ) g -(B 2 ) h -(B 3 ) i -(C 2 ) j -(B 4 ) k –A 2 .

[0550] In some embodiments, the linker is D. In some embodiments, D is an optionally substituted C 1–10 Alkyl, optionally substituted C 2–10 Alkenyl, optionally substituted C 2–10 Alkynyl, optionally substituted C 2–10 Heterocyclic group, optionally substituted C 2–6 Heteroaryl, optionally substituted C 6–12 Aryl, optionally substituted C 2 -C 10 Polyethylene glycol or optionally substituted C 1–10 In some embodiments, D is an optionally substituted C 3 -C 10 cycloalkyl, f is 1, g is 0, h is 0, i is 0, j is 0, and k is 1. In some embodiments, D is an optionally substituted C 3 -C 10 cycloalkyl, f is 1, g is 0, h is 0, i is 0, j is 0, and k is 0. In some embodiments, D is an optionally substituted C 3 -C 10 cycloalkyl, f is 0, g is 0, h is 0, i is 0, j is 0, and k is 1. In some embodiments, D is an optionally substituted C 3 -C 10 cycloalkyl, f is 0, g is 0, h is 0, i is 0, j is 0, and k is 0. In some embodiments, D is an optionally substituted C 3-C 10 carbocyclyl, f is 1, g is 0, h is 0, i is 0, j is 0, and k is 1. In some embodiments, D is an optionally substituted C 3 -C 10 carbocyclyl, f is 1, g is 0, h is 0, i is 0, j is 0, and k is 0. In some embodiments, D is an optionally substituted C 3 -C 10 carbocyclyl, f is 0, g is 0, h is 0, i is 0, j is 0, and k is 1. In some embodiments, D is an optionally substituted C 3 -C 10 carbocyclyl, f is 0, g is 0, h is 0, i is 0, j is 0, and k is 0. In some embodiments, D is:

[0551]

[0552]

[0553]

[0554] In some embodiments, the linker has the following structure:

[0555]

[0556]

[0557]

[0558] In some embodiments, the linker has the structure of Formula III:

[0559] A 1 -(B 1 ) f -(C 1 ) g -(B 2 ) h -(B 3 ) i -(C 2 ) j -(B 4 ) k –A 2

[0560] Formula III

[0561] in

[0562] A 1 is the bond between the linker and the ring system A;

[0563] A 2is the bond between the degradable moiety and the linker;

[0564] B 1 , B 2 , B 3 and B 4 are each independently an optionally substituted ethynyl, an optionally substituted C 6 -C 10 Aryl, optionally substituted C 3 -C 10 Cycloalkyl, optionally substituted C 3 -C 10 Carbocyclic group, optionally substituted C 2 -C 10 Heterocyclic group, optionally substituted C 2 -C 9 Heteroaryl, O, S, S(O) 2 or NR N ;

[0565] Each R N are independently H, optionally substituted C 1–4 Alkyl, optionally substituted C 2–4 Alkenyl, optionally substituted C 2–4 Alkynyl, optionally substituted C 2–10 Heterocyclic group, optionally substituted C 6–12 Aryl or optionally substituted C 1–7 heteroalkyl;

[0566] C 1 and C 2 are each independently carbonyl, thiocarbonyl, sulfonyl or phosphoryl; and

[0567] f, g, h, i, j and k are each independently 0 or 1;

[0568] B is the degradation part;

[0569] Each R 1 are independently halogen, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 Heteroalkyl, optionally substituted C 3 -C 8 Cycloalkyl or optionally substituted C 2 -C 10 Heterocyclic group;

[0570] and

[0571] Each X is independently halogen.

[0572] In some embodiments, the linker has the structure –(L 1 )n -, where n is 1, 2 or 3, and each L 1 O, NR independently N , ethynyl, optionally substituted C 2 -C 10 Heterocyclic group, optionally substituted C 2 -C 9 Heteroaryl, optionally substituted C 6 -C 10 Aryl or optionally substituted C 3 -C 10 Cycloalkyl.

[0573] In some embodiments, at least one L 1 is an optionally substituted C 2 -C 10 In some embodiments, the optionally substituted C 2 -C 10 The heterocyclyl group is a 4-, 5- or 6-membered monocyclic heterocyclyl group. In some embodiments, the 4-, 5- or 6-membered monocyclic heterocyclyl group is:

[0574]

[0575]

[0576] In some embodiments, the optionally substituted C 2 -C 10 The heterocyclic group is a spirocyclic heterocyclic group. In some embodiments, the spirocyclic heterocyclic group is:

[0577]

[0578] In some embodiments, the optionally substituted C 2 -C 10 The heterocyclyl group is a bridged heterocyclyl group. In some embodiments, the bridged heterocyclyl group is:

[0579]

[0580] In some embodiments, the optionally substituted C 2 -C 10 The heterocyclyl is a fused bicyclic heterocyclyl. In some embodiments, the fused bicyclic heterocyclyl is:

[0581]

[0582] In some embodiments, at least one L 1 is an optionally substituted C 2 -C 9 In some embodiments, the linker is -(L 1) q -(optionally substituted C 2 -C 9 Heteroaryl)-(L 1 ) q -, wherein each q is independently 0 or 1. In some embodiments, the optionally substituted C 2 -C 9 Heteroaryl is a 6-membered monocyclic heteroaryl. In some embodiments, the 6-membered monocyclic heteroaryl is:

[0583]

[0584] In some embodiments, at least one L 1 is an optionally substituted C 2 -C 9 In some embodiments, the linker is:

[0585]

[0586] In some embodiments, at least one L 1 is an optionally substituted C 6 -C 10 In some embodiments, the optionally substituted C 6 -C 10 Aryl is a 6-membered monocyclic aryl. In some embodiments, the 6-membered monocyclic aryl is an optionally substituted phenyl.

[0587] In some embodiments, at least one L 1 is an optionally substituted C 3 -C 10 In some embodiments, the optionally substituted C 3 -C 10 Cycloalkyl is a monocyclic cycloalkyl. In some embodiments, the 6-membered monocyclic cycloalkyl is:

[0588]

[0589] In some embodiments, the optionally substituted C 3 -C 10 Cycloalkyl is a bridged cycloalkyl. In some embodiments, the bridged cycloalkyl is:

[0590]

[0591] In some embodiments, at least one L 1 It is ethynyl.

[0592] In some embodiments, one and only one L 1 is O. In some embodiments, one and only one L1 YesNR N In some embodiments, R N is an optionally substituted C 1 -C 4 In some embodiments, R N It's H.

[0593] In some embodiments, the linker has the following structure:

[0594] A 1 -(B 1 ) f -(B 2 ) h -(B 3 ) i -(B 4 ) k –A 2 ,

[0595] Among them B 1 , B 2 , B 3 and B 4 are each independently an optionally substituted ethynyl, an optionally substituted C 6 -C 10 Aryl, optionally substituted C 3 -C 10 Cycloalkyl, optionally substituted C 2 -C 10 Heterocyclic group, optionally substituted C 2 -C 9 Heteroaryl, O or NR N .

[0596] In some embodiments, at least one of f, h, i, and k is 1.

[0597] In some embodiments, B 1 , B 2 , B 3 and B 4 are each independently O, ethynyl, optionally substituted C 2 -C 9 Heteroaryl, optionally substituted C 2 -C 10 Heterocyclic group, optionally substituted C 3 -C 10 Cycloalkyl or optionally substituted C 6 -C 10 In some embodiments, B 1 , B 2 , B 3 and B 4 are each independently an optionally substituted C2 -C 9 Heteroaryl or optionally substituted C 2 -C 10 In some embodiments, B 1 and B 4 Each independently is

[0598]

[0599]

[0600] In some embodiments, B 1 yes:

[0601]

[0602]

[0603] In some embodiments, B 4 yes:

[0604]

[0605]

[0606]

[0607] In some embodiments, B 2 YesNR N In some embodiments, B 2 is NH. In some embodiments, B 2 is an optionally substituted C 2 -C 9 In some embodiments, B 2 yes:

[0608]

[0609] In some embodiments, f is 0. In some embodiments, f is 1. In some embodiments, g is 0. In some embodiments, g is 1. In some embodiments, h is 0. In some embodiments, h is 1. In some embodiments, i is 0. In some embodiments, i is 1. In some embodiments, j is 0. In some embodiments, j is 1. In some embodiments, k is 0. In some embodiments, k is 1.

[0610] In some embodiments, the linker has the structure

[0611]

[0612]

[0613]

[0614] In some embodiments, the shortest chain of atoms connecting two valences of a linker is 2 to 10 atoms long. In some embodiments, the shortest chain of atoms connecting two valences of a linker is 6 atoms long.

[0615] In some embodiments, the linker has any of compounds 1-121 in Table 1 (eg, having a BRG1 IC of at least 5 (eg, at least 7, 10, 15, 20, 25, or 30). 50 With BRM IC 50 In some embodiments, the linker has any one of compounds 1-121 in Table 1 (e.g., having a BRM IC of ++ or better (e.g., +++ or ++++ (e.g., ++++)). 50 In some embodiments, the linker has any one of compounds 1-121 in Table 1 (e.g., having a BRM IC of ++ or better (e.g., +++ or ++++ (e.g., ++++)). 50 and a BRG1 IC of at least 5 (e.g., at least 7, 10, 15, 20, 25, or 30) 50 With BRM IC 50 The structure of the linker in any one of the compounds having a ratio of .

[0616] In one aspect, the invention features a compound selected from 1-121 in Table 1 and a pharmaceutically acceptable salt thereof. In some embodiments, the compound is a BRG1 IC of at least 5 (e.g., at least 7, 10, 15, 20, 25, or 30). 50 With BRMIC 50 Any one of compounds 1-121 in Table 1 or a pharmaceutically acceptable salt thereof having a ratio of . In some embodiments, the compound is a BRM IC having ++ or better (e.g., +++ or ++++ (e.g., ++++)) as seen in Table 15. 50 Any one of compounds 1-121 in Table 1 or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is a BRM IC having ++ or better (e.g., +++ or ++++ (e.g., ++++)) as seen in Table 15. 50 and a BRG1 IC of at least 5 (e.g., at least 7, 10, 15, 20, 25, or 30) 50 With BRM IC 50 Any one of Compounds 1 to 121 in Table 1 or a pharmaceutically acceptable salt thereof.

[0617] In one aspect, the invention features a compound selected from 1-103 in Table 1 and pharmaceutically acceptable salts thereof.

[0618] Table 1. Compounds of the present invention

[0619]

[0620]

[0621]

[0622]

[0623]

[0624]

[0625]

[0626]

[0627]

[0628]

[0629]

[0630]

[0631]

[0632]

[0633]

[0634]

[0635]

[0636]

[0637]

[0638]

[0639]

[0640]

[0641]

[0642]

[0643]

[0644]

[0645]

[0646]

[0647] In some embodiments, the compound has a BRG1 IC of at least 5 50 With BRM IC 50 In some embodiments, the compound has a BRG1 IC of at least 7. 50 With BRM IC 50 In some embodiments, the compound has a BRG1 IC of at least 10. 50 With BRM IC 50 In some embodiments, the compound has a BRG1IC of at least 15. 50 With BRM IC 50 In some embodiments, the compound has a BRG1 IC of at least 20. 50 With BRMIC 50 In some embodiments, the compound has a BRG1 IC of at least 25. 50 With BRM IC 50 In some embodiments, the compound has a BRG1 IC of at least 30. 50 With BRM IC 50 ratio.

[0648] In one aspect, the invention features a pharmaceutical composition including any of the foregoing compounds and a pharmaceutically acceptable excipient.

[0649] In another aspect, the invention features a method of reducing the activity of a BAF complex in a cell, the method involving contacting the cell with an effective amount of any one of the aforementioned compounds or a pharmaceutical composition thereof.

[0650] In some embodiments, the cell is a cancer cell.

[0651] In another aspect, the invention features a method of treating a BAF complex-associated disorder in a subject in need thereof, the method involving administering to the subject an effective amount of any of the aforementioned compounds (e.g., a BRM / BRG1 dual inhibitor compound or a BRM selective compound) or a pharmaceutical composition thereof.

[0652] In some embodiments, the BAF complex-associated disorder is cancer.

[0653] In a further aspect, the invention features a method of inhibiting BRM, the method involving contacting a cell with an effective amount of any of the aforementioned compounds (eg, a BRM / BRG1 dual inhibitor compound or a BRM selective compound) or a pharmaceutical composition thereof.

[0654] In some embodiments, the cell is a cancer cell.

[0655] In another aspect, the invention features a method of inhibiting BRG1, the method involving contacting a cell with an effective amount of any one of the aforementioned compounds or a pharmaceutical composition thereof.

[0656] In some embodiments, the cell is a cancer cell.

[0657] In a further aspect, the invention features a method of inhibiting BRM and BRG1, the method involving contacting a cell with an effective amount of any one of the foregoing compounds or a pharmaceutical composition thereof.

[0658] In some embodiments, the cell is a cancer cell.

[0659] In another aspect, the invention features a method of treating a disorder associated with a BRG1 loss-of-function mutation in a subject in need thereof, the method involving administering to the subject an effective amount of any of the aforementioned compounds (e.g., a BRM / BRG1 dual inhibitor compound or a BRM selective compound) or a pharmaceutical composition thereof.

[0660] In some embodiments, the disorder associated with a BRG1 loss-of-function mutation is cancer. In other embodiments, the subject is determined to have a BRG1 loss-of-function disorder, such as a BRG1 loss-of-function cancer (e.g., the cancer has been determined to include cancer cells with BRG1 loss-of-function).

[0661] In another aspect, the invention features a method of inducing apoptosis in a cell, the method involving contacting the cell with an effective amount of any of the aforementioned compounds (eg, a BRM / BRG1 dual inhibitor compound or a BRM selective compound) or a pharmaceutical composition thereof.

[0662] In some embodiments, the cell is a cancer cell.

[0663] In a further aspect, the invention features a method of treating cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of any of the aforementioned compounds (e.g., a BRM / BRG1 dual inhibitor compound or a BRM selective compound) or a pharmaceutical composition thereof.

[0664] In some embodiments of any of the foregoing methods, the cancer is non-small cell lung cancer, colorectal cancer, bladder cancer, cancer of unknown primary, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, esophageal-gastric cancer, pancreatic cancer, hepatobiliary cancer, soft tissue sarcoma, ovarian cancer, head and neck cancer, renal cell carcinoma, bone cancer, non-Hodgkin lymphoma, small cell lung cancer, prostate cancer, embryonal tumor, germ cell tumor, cervical cancer, thyroid cancer, salivary gland cancer, gastrointestinal neuroendocrine tumors, uterine sarcoma, gastrointestinal stromal tumors, CNS cancer, thymic tumor, adrenocortical carcinoma, appendix cancer, small intestine cancer, or penile cancer.

[0665] In some embodiments of any of the foregoing methods, the cancer is non-small cell lung cancer, colorectal cancer, bladder cancer, cancer of unknown primary, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, or penile cancer.

[0666] In some embodiments of any of the foregoing methods, the cancer is a drug-resistant cancer or has failed to respond to a previous therapy (e.g., vemurafenib, dacarbazine, CTLA4 inhibitors, PD1 inhibitors, interferon therapy, BRAF inhibitors, MEK inhibitors, radiation therapy, temozolomide, irinotecan, CAR-T therapy, Tamoxifen, docetaxel, platinum agents such as carboplatin, taxanes such as paclitaxel and docetaxel, ALK inhibitors, MET inhibitors, Gemcitabine, Neratinib, PARP inhibitors, ARN810, mTOR inhibitors, topotecan, VEGFR2 inhibitors, folate receptor antagonists, demcizumab, fosbretabulin, or PDL1 inhibitors).

[0667] In some embodiments of any of the foregoing methods, the cancer has or has been determined to have 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 determined to have an epidermal growth factor receptor (EGFR) mutation. In some embodiments of any of the foregoing methods, the cancer does not have or has been determined to have an anaplastic lymphoma kinase (ALK) driver mutation. In some embodiments of any of the foregoing methods, the cancer has or has been determined to have 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.

[0668] In another aspect, the present disclosure provides a method for treating a BAF-related disorder (e.g., cancer or viral infection) in a subject in need thereof. The method comprises contacting a cell with an effective amount of any of the aforementioned compounds (e.g., a BRM / BRG1 dual inhibitor compound or a BRM selective compound) or a pharmaceutically acceptable salt thereof, or any of the aforementioned pharmaceutical compositions. In some embodiments, the disorder is a viral infection, which is a retroviridae 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), herpesviridae (e.g., human cytomegalovirus (HCMV), Epstein-Barr virus, herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2), human herpes virus type 6 (HHV-6), herpes virus K*, CMV, varicella-zoster virus), papillomaviridae (e.g., human papillomavirus (HPV, HPV- In some embodiments, the disorder is Coffin-Siris syndrome, neurofibromatosis (e.g., NF-1, NF-2, or schwannomatosis), or multiple meningiomas.

[0669] In another aspect, the present disclosure provides a method for treating a viral infection in a subject in need thereof. The method comprises administering to the subject an effective amount of any one of the aforementioned compounds (e.g., a BRM / BRG1 dual inhibitor compound or a BRM selective compound) or a pharmaceutically acceptable salt thereof, or any one of the aforementioned pharmaceutical compositions. In some embodiments, the viral infection is a retroviridae such as a lentivirus (e.g., human immunodeficiency virus (HIV)) and a delta retrovirus (e.g., human T-cell leukemia virus I (HTLV-I), human T-cell leukemia virus II (HTLV-II)), a hepatoviridae (e.g., hepatitis B virus (HBV)), a flaviviridae (e.g., hepatitis C virus (HCV)), adenoviridae (e.g., human adenovirus), herpesviridae (e.g., human cytomegalovirus (HCMV), Epstein-Barr virus, herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2), human herpes virus type 6 (HHV-6), herpes virus K*, CMV, varicella-zoster virus), a papillomaviridae (e.g., human papillomavirus (HPV, HPV- E1)), infection with a virus of the Parvoviridae family (e.g., Parvovirus B19), the Polyomaviridae family (e.g., JC virus and BK virus), the Paramyxoviridae family (e.g., measles virus), or the Togaviridae family (e.g., Rubella virus).

[0670] In some embodiments of any of the foregoing aspects, the compound is a BRM selective compound. In some embodiments, the BRM selective compound inhibits the level and / or activity of BRM by at least 10 times greater than the level and / or activity of BRG1, and / or the compound binds to BRM by at least 10 times greater than the binding of the compound to BRG1. For example, in some embodiments, the IC of a BRM selective compound is 50 or IP 50 It is the IC of BRG1 50 or IP 50 In some embodiments of any of the foregoing aspects, the compound is a BRM / BRG1 dual inhibitor compound. In some embodiments, the BRM / BRG1 dual inhibitor compound has similar activity against BRM and BRG1 (e.g., the compound has an activity against BRM and BRG1 within 10 times (e.g., less than 5 times, less than 2 times)). In some embodiments, the BRM / BRG1 dual inhibitor compound has greater activity against BRM. In some embodiments, the BRM / BRG1 dual inhibitor compound has greater activity against BRG1. For example, in some embodiments, the IC of a BRM / BRG1 dual inhibitor compound against BRM is 50 or IP 50 In the IC of BRG1 50 or IP 50Within 10 times of.

[0671] In another aspect, the invention features a method of treating melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or a blood cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of any one of the foregoing compounds or a pharmaceutical composition thereof.

[0672] In another aspect, the invention features a method of reducing tumor growth of melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or blood cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of any one of the foregoing compounds or a pharmaceutical composition thereof.

[0673] In another aspect, the invention features a method of inhibiting metastatic progression of melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or a hematological cancer in a subject, the method comprising administering an effective amount of any one of the foregoing compounds or a pharmaceutical composition thereof.

[0674] In another aspect, the invention features a method of inhibiting metastatic colonization of a melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or a hematological cancer in a subject, the method comprising administering an effective amount of any one of the foregoing compounds or a pharmaceutical composition thereof.

[0675] In another aspect, the invention features a method of reducing the level and / or activity of BRG1 and / or BRM in melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or hematological cancer cells, the method comprising contacting the cells with an effective amount of any one of the foregoing compounds or a pharmaceutical composition thereof.

[0676] In some embodiments of any of the above aspects, melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or a hematological cancer cell is in the subject.

[0677] In some embodiments of any of the above aspects, 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 a reference. In some embodiments, the effective amount of the compound reduces the level and / or activity of BRG1 by at least 50% (e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95%) compared to a reference. 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%).

[0678] In some embodiments, an 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 a 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, an 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 a reference for at least 4 days (e.g., 5 days, 6 days, 7 days, 14 days, 28 days or more).

[0679] In some embodiments of any of the above aspects, the effective amount of the compound reduces the level and / or activity of 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 a reference. In some embodiments, the effective amount of the compound reduces the level and / or activity of BRM by at least 50% (e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) compared to a reference. 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%).

[0680] In some embodiments, an effective amount of the compound reduces the level and / or activity of a 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 a 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, an effective amount of the compound reduces the level and / or activity of 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 a reference for at least 4 days (e.g., 5 days, 6 days, 7 days, 14 days, 28 days or more).

[0681] In some embodiments, the subject has cancer. In some embodiments, the cancer expresses BRG1 and / or BRM protein, and / or the cell or subject has been identified as expressing BRG1 and / or BRM. In some embodiments, the cancer expresses BRG1 protein, and / or the cell or subject has been identified as expressing BRG1. In some embodiments, the cancer expresses BRM protein, and / or the cell or subject has been identified as expressing BRM. In some embodiments, the cancer is melanoma (e.g., uveal melanoma, mucosal melanoma, or cutaneous melanoma). In some embodiments, the cancer is prostate cancer. In some embodiments, the cancer is a blood cancer, such as multiple myeloma, large cell lymphoma, acute T-cell leukemia, acute myeloid leukemia, myelodysplastic syndrome, immunoglobulin A lambda 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's 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., cancer has spread to the liver). The metastatic cancer may include cells that exhibit migration and / or invasion of migratory cells, and / or include cells that exhibit endothelial recruitment and / or angiogenesis. In other embodiments, the migratory cancer is a cell migration cancer. In yet other embodiments, the cell migration cancer is a non-metastatic cell migration cancer. The metastatic cancer may be a cancer that spreads by surface seeding of the peritoneum, pleura, pericardium, or subarachnoid space. Alternatively, the metastatic cancer may be a cancer that spreads via the lymphatic system, or a cancer that spreads by the bloodstream. In some embodiments, the effective amount of the agent that reduces the level and / or activity of BRG1 and / or BRM is an amount that effectively inhibits the metastatic colonization of cancer to the liver.

[0682] In some embodiments, the cancer has a mutation in GNAQ. In some embodiments, the cancer has a mutation in GNA11. In some embodiments, the cancer has a mutation in PLCB4. In some embodiments, the cancer has a mutation in CYSLTR2. In some embodiments, the cancer has a mutation in BAP1. In some embodiments, the cancer has a mutation in SF3B1. In some embodiments, the cancer has a mutation in EIF1AX. In some embodiments, the cancer has a TFE3 translocation. In some embodiments, the cancer has a TFEB translocation. In some embodiments, the cancer has a MITF translocation. In some embodiments, the cancer has an EZH2 mutation. In some embodiments, the cancer has a SUZ12 mutation. In some embodiments, the cancer has an EED mutation.

[0683] In some embodiments, the method further comprises administering to the subject an anti-cancer therapy or contacting the cell with an anti-cancer therapy, such as a chemotherapeutic or cytotoxic agent, immunotherapy, surgery, radiotherapy, thermotherapy, or photocoagulation. In some embodiments, the anti-cancer therapy is a chemotherapeutic or cytotoxic agent, such as an antimetabolite, an antimitotic agent, an antitumor antibiotic, an asparagine-specific enzyme, a bisphosphonate, an antitumor agent, an alkylating agent, a DNA repair enzyme inhibitor, a histone deacetylase inhibitor, a corticosteroid, a demethylating agent, an immunomodulator, a janus-related kinase inhibitor, a phosphatidylinositol 3-kinase inhibitor, a proteasome inhibitor, or a tyrosine kinase inhibitor.

[0684] In some embodiments, the compounds of the invention are used in combination with another anticancer therapy for treating uveal melanoma, such as surgery, a MEK inhibitor, and / or a PKC inhibitor. For example, in some embodiments, the method further comprises performing surgery before, after, or simultaneously with the administration of the compounds of the invention. In some embodiments, the method further comprises administering a MEK inhibitor and / or a PKC inhibitor before, after, or simultaneously with the administration of the compounds of the invention.

[0685] In some embodiments, the anti-cancer therapy and the compound of the invention are administered within 28 days of each other, and the respective amounts are together effective to treat the subject.

[0686] In some embodiments, the subject or cancer has and / or has been identified as having a BRG1 loss-of-function mutation.

[0687] In some embodiments, the cancer is resistant to one or more chemotherapeutic or cytotoxic agents (e.g., the cancer has been identified as resistant to chemotherapeutic or cytotoxic agents, such as by a genetic marker, or is potentially resistant to chemotherapeutic or cytotoxic agents, such as a cancer that fails to respond to chemotherapeutic or cytotoxic agents). In some embodiments, the cancer fails to respond to one or more chemotherapeutic or cytotoxic agents. In some embodiments, the cancer is resistant or fails to respond to dacarbazine, temozolomide, cisplatin, treosulfan, fotemustine, IMCgp100, a CTLA-4 inhibitor (e.g., ipilimumab), a PD-1 inhibitor (e.g., nivolumab or pembrolizumab), a PD-L1 inhibitor (e.g., atezolizumab, avelumab, or durvalumab), a mitogen-activated protein kinase (MEK) inhibitor (e.g., selumetinib, binumetinib, or trametinib), and / or a protein kinase C (PKC) inhibitor (e.g., sotrastaurin or IDE196).

[0688] In some embodiments, the cancer is resistant or unresponsive to a previously administered therapeutic agent for treating uveal melanoma, such as a MEK inhibitor or a PKC inhibitor. For example, in some embodiments, the cancer is resistant or unresponsive to a mitogen-activated protein kinase (MEK) inhibitor (e.g., selumetinib, bimetinib, or trametinib) and / or a protein kinase C (PKC) inhibitor (e.g., sotrastaurin or IDE196).

[0689] In one aspect, the present invention provides a compound disclosed herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition disclosed herein, for use in therapy.

[0690] In one aspect, the present invention provides a compound disclosed herein or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition disclosed herein, for use in reducing the activity of a BAF complex in a cell.

[0691] In some embodiments, the BAF complex is in a cancer cell.

[0692] In one aspect, the present invention provides a compound disclosed herein or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition disclosed herein, for use in treating a BAF complex-related disorder.

[0693] In some embodiments, the BAF complex-associated disorder is cancer or a viral infection.

[0694] In one aspect, the present invention provides a compound disclosed herein or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition disclosed herein, for use in inhibiting a BRM in a cell.

[0695] In some embodiments, the cell is a cancer cell.

[0696] In one aspect, the present invention provides a compound disclosed herein or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition disclosed herein, for use in treating a disorder associated with a BRG1 loss-of-function mutation.

[0697] In some embodiments, the disorder associated with a BRG1 loss-of-function mutation is cancer.

[0698] In one aspect, the present invention provides a compound disclosed herein or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition disclosed herein, for use in inducing apoptosis in a cell.

[0699] In some embodiments, the cell is a cancer cell.

[0700] In one aspect, the present invention provides a compound disclosed herein or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition disclosed herein, for use in treating cancer.

[0701] In some embodiments, the cancer is non-small cell lung cancer, colorectal cancer, bladder cancer, cancer of unknown primary, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, esophageal-gastric cancer, pancreatic cancer, hepatobiliary cancer, soft tissue sarcoma, ovarian cancer, head and neck cancer, renal cell carcinoma, bone cancer, non-Hodgkin 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 intestine cancer, or penile cancer.

[0702] In some embodiments, the cancer is non-small cell lung cancer, colorectal cancer, bladder cancer, cancer of unknown primary, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, or penile cancer.

[0703] In some embodiments, the cancer is non-small cell lung cancer.In some embodiments, the cancer is soft tissue sarcoma.

[0704] In one aspect, the present invention provides a compound disclosed herein or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition disclosed herein, for use in treating a cancer selected from melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, and blood cancer.

[0705] In one aspect, the present invention provides a compound disclosed herein or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition disclosed herein, for use in reducing tumor growth of a cancer selected from melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, and blood cancer.

[0706] In one aspect, the present invention provides a compound disclosed herein or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition disclosed herein, for use in inhibiting metastatic progression of a cancer selected from melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, and blood cancer.

[0707] In one aspect, the present invention provides a compound disclosed herein or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition disclosed herein, for use in inhibiting metastatic colonization of a cancer selected from melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, and hematological cancer.

[0708] In one aspect, the present invention provides a compound disclosed herein or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition disclosed herein, for use in reducing the level and / or activity of a BRM in a cancer cell selected from melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, and a blood cancer.

[0709] In some embodiments, the cell is in a subject.

[0710] In some embodiments, the cancer is metastatic.

[0711] In some embodiments, the use further comprises anti-cancer therapy.

[0712] In some embodiments, the anti-cancer therapy is a chemotherapeutic or cytotoxic agent, immunotherapy, surgery, radiation therapy, thermotherapy, or photocoagulation.

[0713] In some embodiments, the anti-cancer therapy is surgery.

[0714] In some embodiments, the anti-cancer therapy is a chemotherapeutic agent or a cytotoxic agent.

[0715] In some embodiments, the chemotherapeutic or cytotoxic agent is an antimetabolite, an antimitotic agent, an antitumor antibiotic, an asparagine-specific enzyme, a bisphosphonate, an antineoplastic agent, an alkylating agent, a DNA repair enzyme inhibitor, a histone deacetylase inhibitor, a corticosteroid, a demethylating agent, an immunomodulator, a janus-related kinase inhibitor, a phosphatidylinositol 3-kinase inhibitor, a proteasome inhibitor, or a tyrosine kinase inhibitor.

[0716] In some embodiments, the one or more chemotherapeutic or cytotoxic agents are dacarbazine, temozolomide, cisplatin, trosulfan, fotemustine, IMCgp100, CTLA-4 inhibitors, PD-1 inhibitors, PD-L1 inhibitors, mitogen-activated protein kinase inhibitors, and / or protein kinase C inhibitors.

[0717] In some embodiments, the anti-cancer therapy and the compound or pharmaceutical composition thereof are administered within 28 days of each other, and the respective amounts are together effective to treat the subject.

[0718] In some embodiments, the subject or cancer has and / or has been identified as having a BRG1 loss-of-function mutation.

[0719] In some embodiments, the cancer has failed to respond or has progressed following administration of one or more chemotherapeutic or cytotoxic agents.

[0720] In some embodiments, the cancer is resistant or predicted to be resistant to one or more chemotherapeutic agents.

[0721] In some embodiments, the one or more chemotherapeutic or cytotoxic agents are dacarbazine, temozolomide, cisplatin, trosulfan, fotemustine, IMCgp100, CTLA-4 inhibitors, PD-1 inhibitors, PD-L1 inhibitors, mitogen-activated protein kinase inhibitors, and / or protein kinase C inhibitors.

[0722] In some embodiments, the cancer is melanoma. In some embodiments, the melanoma is uveal melanoma. In some embodiments, the melanoma is mucosal melanoma. In some embodiments, the melanoma is cutaneous melanoma. In some embodiments, the cancer is a blood cancer. In some embodiments, the blood cancer is multiple myeloma, large cell lymphoma, acute T-cell leukemia, acute myeloid leukemia, myelodysplastic syndrome, immunoglobulin A lambda myeloma, diffuse mixed histiocytic and lymphocytic lymphoma, B-cell lymphoma, acute lymphoblastic leukemia, diffuse large cell lymphoma or non-Hodgkin lymphoma. In some embodiments, the cancer is prostate cancer. In some embodiments, the cancer is breast cancer. In some embodiments, the breast cancer is 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. In some embodiments, the bone cancer is Ewing's sarcoma. In some embodiments, the cancer is renal cell carcinoma. In some embodiments, the renal cell carcinoma is a microphthalmia transcription factor (MITF) family translocation renal cell carcinoma.

[0723] In one aspect, the present invention provides a compound disclosed herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition disclosed herein for use in treating a viral infection.

[0724] In some embodiments, the viral infection is a Retroviridae, Hepadnaviridae, Flaviviridae, Adenoviridae, Herpesviridae, Papillomaviridae, Parvoviridae, Polyomaviridae, Paramyxoviridae, or Togaviridae viral infection.

[0725] In one aspect, the present invention provides the use of any of the aforementioned compounds (eg, BRM / BRG1 dual inhibitor compounds or BRM selective compounds) or their pharmaceutically acceptable salts or any of the aforementioned pharmaceutical compositions in the manufacture of a medicament. In some embodiments, the use is as described for the methods described herein.

[0726] Chemical terms

[0727] The terminology used herein is for the purpose of describing particular embodiments and is not intended to be limiting.

[0728] For any of the following chemical definitions, the number following the atomic symbol indicates the total number of atoms of that element present in the particular chemical moiety. As will be appreciated, other atoms (such as H atoms) or substituents as described herein may be present as necessary to satisfy the valence of the atom. For example, unsubstituted C 2 Alkyl groups have the formula –CH 2 CH 3 When used for groups defined herein, references to the number of carbon atoms include the divalent carbon in acetal and ketal groups, but exclude the carbonyl carbon in acyl, ester, carbonate or carbamate groups. References to the number of oxygen, nitrogen or sulfur atoms in a heteroaryl group include only those atoms that form part of the heterocyclic ring.

[0729] As used herein, term " acyl " represents H or alkyl connected to the parent molecular group via carbonyl as defined herein, such as formyl (i.e., formaldehyde), acetyl, trifluoroacetyl, propionyl and butyryl. Exemplary unsubstituted acyl includes 1 to 6, 1 to 11, or 1 to 21 carbons.

[0730] The term "alkyl" as used herein refers to a branched or straight-chain monovalent saturated aliphatic hydrocarbon group of 1 to 20 carbon atoms (eg, 1 to 16 carbon atoms, 1 to 10 carbon atoms, 1 to 6 carbon atoms, or 1 to 3 carbon atoms).

[0731] Alkylene is a divalent alkyl group. The term "alkenyl" as used herein, alone or in combination with other groups, refers to a straight or branched hydrocarbon residue having a carbon-carbon double bond and having 2 to 20 carbon atoms (e.g., 2 to 16 carbon atoms, 2 to 10 carbon atoms, 2 to 6 carbon atoms, or 2 carbon atoms).

[0732] The term "alkynyl" as used herein, alone or in combination with other groups, refers to a straight or branched hydrocarbon residue having a carbon-carbon triple bond and having 2 to 20 carbon atoms (e.g., 2 to 16 carbon atoms, 2 to 10 carbon atoms, 2 to 6 carbon atoms, or 2 carbon atoms).

[0733] As used herein, the term "amino" refers to -N(R N1 ) 2 , where each RN1 H, OH, NO independently 2 、N(R N2 ) 2 、SO 2 OR N2 、SO 2 R N2 , SOR N2 , N-protecting group, alkyl, alkoxy, aryl, aralkyl, cycloalkyl, acyl (e.g., acetyl, trifluoroacetyl, or other acyl described herein), wherein these listed R N1 Each of the groups may be optionally substituted; or two R N1 to form an alkylene group or a heteroalkylene group, and wherein each R N2 are independently H, alkyl or aryl. The amino group of the present invention may be an unsubstituted amino group (i.e., -NH 2 ) or substituted amino (i.e. –N(R N1 ) 2 ).

[0734] The term "aryl" as used herein refers to an aromatic mono- or polycarbocyclic group of 6 to 12 carbon atoms having at least one aromatic ring. In the case of polycyclics, the aryl contains 2 or 3 rings. Examples of such groups include, but are not limited to, phenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, 1,2-dihydronaphthyl, indanyl and 1H-indenyl.

[0735] As used herein, the term "aralkyl" refers to an alkyl group substituted with an aryl group. Unsubstituted aralkyl groups contain 7 to 30 carbon atoms (e.g., 7 to 16 or 7 to 20 carbon atoms, such as C 1 -C 6 Alkyl C 6 -C 10 Aryl, C 1 -C 10 Alkyl C 6 -C 10 Aryl, or C 1 -C 20 Alkyl C 6 -C 10 In some embodiments, the alkyl and aryl groups are each further substituted with 1, 2, 3, or 4 substituents as defined herein for the corresponding groups, depending on valency permitting.

[0736] The term "azido" as used herein represents -N 3 Group.

[0737] The term "bridged polycycloalkyl" as used herein refers to a bridged polycyclic group of 5 to 20 carbons containing 1 to 3 bridges. The bridged polycycloalkyl may be unsubstituted or substituted as defined herein for cycloalkyl.

[0738] The term "cyano" as used herein represents a -CN group.

[0739] As used herein, the term "carbocyclyl" refers to a non-aromatic C 3 -C 12 Monocyclic, bicyclic or tricyclic structures. Carbocyclic structures include cycloalkyl and unsaturated carbocyclic groups.

[0740] As used herein, the term "cycloalkyl" refers to a saturated, non-aromatic, monovalent, monocyclic, bicyclic or tricyclic radical of 3 to 10, preferably 3 to 6 carbon atoms. A cycloalkyl radical may be fully saturated, or contain one or more double or triple bonds, provided that no ring is aromatic. The term is further exemplified by radicals such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl and adamantyl. As used herein, the term "CH 2 "-cycloalkyl" refers to cycloalkyl-CH 2 - groups (e.g. cyclopropylmethyl and cyclobutylmethyl).

[0741] As used herein, the term "halo" refers to a fluorine (fluoro), chlorine (chloro), bromine (bromo) or iodine (iodo) group.

[0742] As used herein, the term "heteroalkyl" refers to an alkyl group as defined herein in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen or sulfur. In some embodiments, the heteroalkyl group is further substituted by 1, 2, 3 or 4 substituents as described herein for the alkyl group. An example of a heteroalkyl group is an "alkoxy group", which refers to an alkyl group-O- (e.g., methoxy and ethoxy) as used herein. A heteroalkylene group is a divalent heteroalkyl group. As used herein, the term "heteroalkenyl" refers to an alkenyl group as defined herein in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen or sulfur. In some embodiments, as valence permits, a heteroalkenyl group is further substituted by 1, 2, 3 or 4 substituents as described herein for the alkenyl group. An example of a heteroalkenyl group is an "alkenyloxy group", which refers to an alkenyl group-O- as used herein. A heteroalkenylene group is a divalent heteroalkenyl group. As used herein, the term "heteroalkynyl" refers to an alkynyl group as defined herein in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen or sulfur. In some embodiments, the heteroalkynyl group is further substituted with 1, 2, 3, or 4 substituents as described herein for alkynyl groups, as valency permits. An example of a heteroalkynyl group is "alkynyloxy," which as used herein refers to alkynyl-O-. A heteroalkynylene group is a divalent heteroalkynyl group.

[0743] As used herein, the term "heteroaryl" refers to a monocyclic, bicyclic or tricyclic group of 5 to 12 atoms having at least one aromatic ring and containing 1, 2 or 3 ring atoms selected from nitrogen, oxygen and sulphur and the remaining ring atoms being carbon. One or two ring carbon atoms of the heteroaryl can be replaced by a carbonyl group. Examples of heteroaryl are pyridyl, pyrazolyl, benzoxazolyl, benzimidazolyl, benzothiazolyl, imidazolyl, oxazolyl and thiazolyl.

[0744] As used herein, the term "heteroarylalkyl" refers to an alkyl group substituted with a heteroaryl group. Unsubstituted heteroarylalkyl groups contain 7 to 30 carbon atoms (e.g., 7 to 16 or 7 to 20 carbon atoms, such as C 1 -C 6 Alkyl C 2 -C 9 Heteroaryl, C 1 -C 10 Alkyl C 2 -C 9 Heteroaryl, or C 1 -C 20 Alkyl C 2 -C 9 In some embodiments, alkyl and heteroaryl are each further substituted with 1, 2, 3, or 4 substituents as defined herein for the corresponding groups, as valency permits.

[0745] As used herein, the term "heterocyclyl" refers to a monocyclic, bicyclic or tricyclic group having 3 to 12 atoms, having at least one ring containing 1, 2, 3 or 4 ring atoms selected from N, O or S, wherein no ring is aromatic. Examples of heterocyclyl include, but are not limited to, morpholinyl, thiomorpholinyl, furanyl, piperazinyl, piperidinyl, pyranyl, pyrrolidinyl, tetrahydropyranyl, tetrahydrofuranyl and 1,3-dioxanyl.

[0746] As used herein, the term "heterocyclylalkyl" represents an alkyl group substituted with a heterocyclyl group. Unsubstituted heterocyclylalkyl contains 7 to 30 carbon atoms (e.g., 7 to 16 or 7 to 20 carbon atoms, such as C 1 -C 6 Alkyl C 2 -C 9 Heterocyclic group, C 1 -C 10 Alkyl C 2 -C 9 Heterocyclic group, or C 1 -C 20 Alkyl C 2 -C 9 In some embodiments, alkyl and heterocyclyl are each further substituted with 1, 2, 3, or 4 substituents as defined herein for the corresponding groups.

[0747] The term "hydroxyalkyl" as used herein represents an alkyl group substituted with an -OH group.

[0748] The term "hydroxyl" as used herein represents an -OH group.

[0749] The term "N-protecting group" as used herein represents those groups intended to protect an amino group from undesirable reactions during synthetic procedures. 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, aroyl or carbamoyl groups such as formyl, acetyl, propionyl, pivaloyl, 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-containing groups such as benzenesulfonyl and p-toluenesulfonyl; carbamate-forming groups such as benzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 3, 5-dimethoxybenzyloxycarbonyl, 2,4-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, isopropyloxycarbonyl, ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl, cyclopentyloxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl and phenylthiocarbonyl, arylalkyl groups such as benzyl, triphenylmethyl and benzyloxymethyl, and silyl groups such as trimethylsilyl. Preferred N-protecting groups are allyloxycarbonyl (alloc), formyl, acetyl, benzoyl, pivaloyl, tert-butylacetyl, alanyl, phenylsulfonyl, benzyl, tert-butoxycarbonyl (Boc) and benzyloxycarbonyl (Cbz).

[0750] As used herein, the term "nitro" represents -NO 2 Group.

[0751] As used herein, the term "oxo" represents a divalent oxygen atom (e.g., the structure of oxo can be shown as =O). For example, a carbonyl group is a carbon (e.g., an alkyl carbon, an alkenyl carbon, an alkynyl carbon, a heteroalkyl carbon, a heteroalkenyl carbon, a heteroalkynyl carbon, a carbocyclyl carbon, etc.) substituted with oxo. Alternatively, sulfur can be substituted with one or two oxo groups (e.g., -SO- or -SO- in a substituted heteroalkyl, heteroalkenyl, heteroalkynyl, or heterocyclyl group). 2 -).

[0752] The term "thiol" as used herein represents a -SH group.

[0753] Alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl (e.g., cycloalkyl), aryl, heteroaryl, and heterocyclyl can be substituted or unsubstituted. When substituted, unless otherwise indicated, 1, 2, 3, 4, or 5 substituents will be present as valence permits. The 1 to 5 substituents are each independently selected from acyl, alkyl (e.g., unsubstituted and substituted, wherein the substituents include any of the groups described herein, such as aryl, halogen, hydroxy), alkenyl, alkynyl, aryl (e.g., substituted and unsubstituted phenyl), carbocyclyl (e.g., substituted and unsubstituted cycloalkyl), halogen (e.g., fluorine), hydroxy, heteroalkyl (e.g., substituted and unsubstituted methoxy, ethoxy, or thioalkoxy), heteroalkenyl, heteroalkynyl, heteroaryl, heterocyclyl, amino (e.g., NH 2 or mono- or di-alkylamino), azido, cyano, nitro, thiol, and oxo. Each substituent is unsubstituted or substituted with an unsubstituted substituent as defined herein for each corresponding group. In some embodiments, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl are optionally substituted with 1, 2, 3, 4, or 5 independently selected from aryl (e.g., substituted and unsubstituted phenyl), carbocyclyl (e.g., substituted and unsubstituted cycloalkyl), halogen (e.g., fluorine), hydroxy, heteroaryl, heterocyclyl, amino (e.g., NH 2 In some embodiments, the substituents are substituted with substituents selected from the group consisting of alkyl, alkyl, styryl, thiol, alkylamino ...

[0754] The compounds of the present invention may have one or more asymmetric carbon atoms, and may exist in the form of an optically pure enantiomer, a mixture of enantiomers such as a racemate, an optically pure diastereomer, a mixture of diastereomers, a diastereomeric racemate, or a mixture of diastereomeric racemates. The optically active form may be obtained, for example, by resolving the racemate, by asymmetric synthesis or asymmetric chromatography (chromatography using a chiral adsorbent or eluent). That is, some disclosed compounds may exist in various stereoisomeric forms. Stereoisomers are compounds that differ only in their spatial arrangement. Enantiomers are stereoisomer pairs whose mirror images are non-superimposable, and the most common reason is that they contain asymmetrically substituted carbon atoms that serve as chiral centers. "Enantiomers" refer to one of a pair of molecules that are mirror images and non-superimposable. Diastereomers are stereoisomers that are not related as mirror images, and the most common reason is that they contain two or more asymmetrically substituted carbon atoms, and represent the configuration of substituents around one or more chiral carbon atoms. The enantiomers of the compound can be prepared, for example, by separating the enantiomers from the racemate using one or more well-known techniques and methods (e.g., chiral chromatography and separation methods based thereon). One skilled in the art can easily determine the appropriate techniques and / or methods for separating the enantiomers of the compounds described herein from the racemic mixture. "Racemate" or "racemic mixture" refers to a compound containing two enantiomers, wherein such a mixture does not exhibit optical activity; i.e., they do not rotate the plane of polarized light. "Geometric isomers" refers to isomers with different orientations of substituent atoms associated with a carbon-carbon double bond, a cycloalkyl ring, or a bridged bicyclic system. The atoms (non-H) on each side of the carbon-carbon double bond can be in E (substituents on opposite sides of the carbon-carbon double bond) or Z (substituents oriented on the same side) configuration. "R", "S", "S*", "R*", "E", "Z", "cis" and "trans" indicate configurations relative to the core molecule. Certain disclosed compounds may exist in atropisomeric forms. Atropisomers are stereoisomers resulting from hindered rotation about a single bond, where the steric strain barrier to rotation is high enough to separate the conformers. The compounds of the present invention can be prepared as single isomers by isomer-specific synthesis or resolution from an isomeric mixture. Conventional resolution techniques include forming salts of the free bases of each isomer of an isomeric pair using an optically active acid (followed by fractional crystallization and regeneration of the free base), forming salts of the acid form of each isomer of an isomeric pair using an optically active amine (followed by fractional crystallization and regeneration of the free acid), forming esters or amides of each isomer of an isomeric pair using an optically pure acid, amine or alcohol (followed by chromatographic separation and removal of the chiral auxiliary), or resolving isomeric mixtures of the starting material or final product using various well-known chromatographic methods.When the stereochemistry of a disclosed compound is named or depicted by structure, the stereoisomer named or depicted is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by weight relative to the other stereoisomers. When a single enantiomer is named or depicted by structure, the enantiomer depicted or named is at least 60%, 70%, 80%, 90%, 99%, or 99.9% optically pure. When a single diastereomer is named or depicted by structure, the diastereomer depicted or named is at least 60%, 70%, 80%, 90%, 99%, or 99.9% pure by weight. The optical purity percentage is the ratio of the weight of an enantiomer to the weight of the enantiomer plus the weight of its optical isomers. Diastereomeric 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 depicted by structure, the named or depicted stereoisomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% pure by mole fraction relative to the other stereoisomers. When a single enantiomer is named or depicted by structure, the depicted or named enantiomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% pure by mole fraction. When a single diastereomer is named or depicted by structure, the depicted or named diastereomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% pure by mole fraction. The percent purity by mole fraction is the ratio of the number of moles of an enantiomer to the number of moles of that enantiomer plus the number of moles of its optical isomers. Similarly, the percent purity on a molar basis is the ratio of the number of moles of a diastereomer to the number of moles of that diastereomer plus the number of moles of its isomers. When a disclosed compound is named or depicted by a structure without indicating stereochemistry and the compound has at least one chiral center, it is understood that the name or structure encompasses either enantiomer of the compound free of the corresponding optical isomer, a racemic mixture of the compound, or a mixture enriched in one enantiomer relative to its corresponding optical isomer. When a disclosed compound is named or depicted by structure without indicating stereochemistry and has two or more chiral centers, it is understood that the name or structure encompasses one diastereomer free of other diastereomers, a plurality of diastereomers free of other diastereomeric pairs, a mixture of diastereomers, a mixture of diastereomeric pairs, a diastereomeric mixture in which one diastereomer is enriched relative to the other diastereomers, or a diastereomeric mixture in which one or more diastereomers are enriched relative to the other diastereomers. The present invention encompasses all of these forms.

[0755] Compounds of the present disclosure also include all isotopes of atoms that occur in intermediates or final compounds. "Isotopes" refer to atoms with the same atomic number but different mass numbers due to different numbers of neutrons in the nucleus. For example, isotopes of hydrogen include tritium and deuterium.

[0756] Unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. Exemplary isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, such as 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 C-labeled) can be used in compound or substrate tissue distribution assays. Due to their ease of preparation and detectability, tritiated (i.e. 3 H) and carbon-14 (i.e. 14 C) isotopes may be useful. In addition, heavier isotopes such as deuterium (i.e. 2 H) substitution may provide certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements). In some embodiments, one or more hydrogen atoms are replaced by 2 H or 3 H is replaced, or one or more carbon atoms are 13 C- or 14 C-enriched carbon substitution. Positron emitting isotopes such as 15 O. 13 N. 11 C and 18F can be used for positron emission tomography (PET) studies to examine substrate receptor occupancy. The preparation of isotope-labeled compounds is known to those skilled in the art. For example, isotope-labeled compounds can generally be prepared by replacing non-isotope-labeled reagents with isotope-labeled reagents following procedures similar to those disclosed for the compounds of the present invention described herein. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as commonly understood by those of ordinary skill in the art to which the present invention belongs. Methods and materials for the present disclosure are described herein; other suitable methods and materials known in the art may also be used. These materials, methods and examples are exemplary only and are not intended to be 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 a conflict, this specification (including definitions) shall prevail.

[0757] definition

[0758] In this application, unless the context clearly indicates otherwise, (i) the term "a" or "an" may be understood to mean "at least one"; (ii) the term "or" may be understood to mean "and / or"; and (iii) the terms "comprising" and "including" may be understood to cover the components or steps listed itemized, whether presented alone or together with one or more additional components or steps.

[0759] As used herein, the terms "about" and "approximately" refer to values ​​within 10% above or below the value being described. For example, the term "about 5 nM" indicates a range of 4.5 to 5.5 nM.

[0760] The term "administering" as used herein refers to administering a composition (e.g., a compound as described herein or a formulation comprising a compound as described herein) to a subject or system. Administration to an animal subject (e.g., to a human) can be by any appropriate route. For example, in some embodiments, administration can be intrabronchial (including by bronchial instillation), buccal, enteral, intradermal, intraarterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intratumoral, intravenous, intraventricular, transmucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (including by intratracheal instillation), transdermal, vaginal, and vitreous.

[0761] The term "BAF complex" as used herein refers to a BRG1- or HRBM-associated factor complex in human cells.

[0762] As used herein, the term "BAF complex-associated disorder" refers to a disorder caused or affected by the activity level of the BAF complex.

[0763] As used herein, the term "BRG1 loss-of-function mutation" refers to a mutation in BRG1 that results in a decrease in the activity of the protein (e.g., a decrease in BRG1 activity of at least 1%, e.g., a decrease in BRG1 activity of 2%, 5%, 10%, 25%, 50%, or 100%). Exemplary BRG1 loss-of-function mutations include, but are not limited to, homozygous BRG1 mutations and deletions at the C-terminus of BRG1.

[0764] As used herein, the term "BRG1 loss-of-function disorder" refers to a disorder (e.g., cancer) that exhibits reduced BRG1 activity (e.g., at least 1% reduced BRG1 activity, e.g., 2%, 5%, 10%, 25%, 50% or 100% reduced BRG1 activity).

[0765] The term "cancer" refers to conditions caused by the proliferation of malignant neoplastic cells, such as tumors, neoplasms, carcinomas, sarcomas, leukemias and lymphomas.

[0766] As used herein, "combination therapy" or "combination administration" refers to the administration of 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 cycle of each agent so that the effects of a single agent on the subject overlap. In some embodiments, the delivery of two or more agents is simultaneous or concurrent, and these agents can be co-formulated. In some embodiments, two or more agents are not co-formulated, and are sequentially administered as part of a prescribed regimen. In some embodiments, the combined administration of two or more agents or treatments results in a greater reduction in the symptoms or other parameters associated with the disorder than observed in the delivery of a single agent or treatment or in the absence of another agent or treatment. The effects of the two treatments can be partially cumulative, completely cumulative, or greater than cumulative (e.g., synergistic). The sequential or substantially simultaneous administration of each therapeutic agent can be achieved by any appropriate route, including but not limited to oral routes, intravenous routes, intramuscular routes, and direct absorption by mucosal tissues. Therapeutic agents can be administered by the same route or by different routes. For example, a first therapeutic agent of the combination may be administered by intravenous injection, while a second therapeutic agent of the combination may be administered orally.

[0767] "Determining the level of a protein or RNA" refers to detecting a protein or RNA directly or indirectly by methods known in the art. "Direct determination" refers to performing a process (e.g., performing an assay or test on a sample, or "analyzing a sample," as the term is defined herein) to obtain a physical entity or value. "Indirect determination" refers to receiving a physical entity or value from another party or source (e.g., a third-party laboratory that directly obtains a 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, immunohistochemical analysis, matrix-assisted laser desorption / ionization time of flight (MALDI-TOF) mass spectrometry, liquid chromatography (LC)-mass spectrometry, microcytometry, microscopy, fluorescence activated cell sorting (FACS) and flow cytometry, as well as assays based on protein properties (including but not limited to enzyme activity or interactions with other protein partners). Methods of measuring RNA levels are known in the art and include, but are not limited to, quantitative polymerase chain reaction (qPCR) and Northern blot analysis.

[0768] "Reducing the activity of the BAF complex" refers to reducing the level of activity associated with the BAF complex or an associated downstream effect. 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 the method described in Kadoch et al., Cell, 2013, 153, 71-85, which method is incorporated herein by reference.

[0769] The term "degrader" as used herein refers to a small molecule compound that includes a degradation moiety, wherein the compound interacts with a protein (e.g., BRG1 and / or BRM) in a manner that results in degradation of the protein, e.g., binding of the compound results in a decrease in the level of the protein in, e.g., a cell or subject, by at least 5%.

[0770] As used herein, the term "degradation moiety" refers to a moiety whose binding results in degradation of a protein, such as BRG1 and / or BRM. In one example, the moiety binds to a protease or ubiquitin ligase that metabolizes the protein, such as BRG1 and / or BRM.

[0771] "Modulating the activity of a BAF complex" refers to changing the level of activity associated with a BAF complex (e.g., GBAF) or an associated downstream effect. The level of activity of a BAF complex can be measured using any method known in the art, such as the method described in Kadoch et al., Cell 153:71-85 (2013), which is incorporated herein by reference.

[0772] "Reducing the activity of BRG1 and / or BRM" refers to reducing the activity level associated with BRG1 and / or BRM or the associated downstream effects. A non-limiting example of inhibiting the activity of BRG1 and / or BRM is reducing the level of BAF complex in a cell. The activity level of BRG1 and / or BRM can be measured using any method known in the art. In some embodiments, the agent that reduces the activity of BRG1 and / or BRM is a small molecule BRG1 and / or BRM degrader.

[0773] "Decreasing the level of BRG1 and / or BRM" refers to decreasing the level of BRG1 and / or BRM in a cell or a subject. The level of BRG1 and / or BRM can be measured using any method known in the art.

[0774] "Level" refers to the level of a protein or mRNA encoding the protein compared to a reference. A reference can be any available reference as defined herein. A "reduced level" or "increased level" of a protein refers to a reduction or increase in protein levels compared to a reference (e.g., a reduction or increase of about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500% or more; a reduction or increase of more than about 1% compared to a reference. The level of a protein can be expressed as a percentage of mass / volume (e.g., g / dL, mg / mL, μg / mL, ng / mL) or relative to the total protein or mRNA in the sample.

[0775] As used herein, the term "inhibit BRM" refers to blocking or reducing the level or activity of the ATPase catalytic binding domain or bromodomain of a protein. BRM inhibition can be determined using methods known in the art, such as BRM ATPase assays, Nano DSF assays, or BRM luciferase cell assays.

[0776] The term "pharmaceutical composition" as used herein represents a composition containing a compound as described herein formulated with a pharmaceutically acceptable excipient and suitable for administration to a mammal, such as a human. Typically, a pharmaceutical composition is manufactured or sold under the approval of a governmental regulatory agency as part of a therapeutic regimen for treating a mammalian disease. The pharmaceutical composition can be formulated, for example, for oral administration in a unit dosage form (e.g., tablets, capsules, caplets, gel caps, or syrups); for topical administration (e.g., as a cream, gel, lotion, or ointment); for intravenous administration (e.g., as a sterile solution in a solvent system suitable for intravenous use without particulate plugs); or in any other pharmaceutically acceptable formulation.

[0777] As used herein, "pharmaceutically acceptable excipients" refers to any ingredient other than the compounds described herein (e.g., a vehicle capable of suspending or dissolving the active compound) and has substantially non-toxic and non-inflammatory properties to the patient. Excipients may include, for example, anti-adherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colorants), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavoring agents, fragrances, glidants (flow promoters), lubricants, preservatives, printing inks, sorbents, suspending or dispersing agents, sweeteners, and hydration water.

[0778] As used herein, the term "pharmaceutically acceptable salt" refers to any pharmaceutically acceptable salt of a compound (e.g., any compound of Formula I or II). Pharmaceutically acceptable salts of any compound described herein may include those suitable for use in contact with human and animal tissues without undue toxicity, irritation, allergic reactions, and commensurate with a reasonable benefit / risk ratio within the scope of reasonable medical judgment. 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, (ed. P. H. Stahl and C. G. Wermuth), Wiley-VCH, 2008. The salt can be prepared in situ during the final separation and purification of the compounds described herein, or prepared separately by reacting the free base group with a suitable organic acid.

[0779] The compounds of the present invention may have ionizable groups so as to be prepared into pharmaceutically acceptable salts. These salts may be acid addition salts involving inorganic or organic acids, or in the case of acidic forms of the compounds of the present invention, the salts may be prepared from inorganic or organic bases. The compounds are often prepared or used as pharmaceutically acceptable salts 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.

[0780] "Reference" refers to any available reference for comparing protein or RNA levels. A reference can be any sample, standard, standard curve or level for comparison purposes. A reference can be a normal reference sample or 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 a normal cell or normal tissue; a sample (such as a cell or tissue) from a subject without a 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 treated with the compounds of the present invention; or a sample of a purified protein or RNA (such as any described herein) at a known normal concentration. "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 a healthy control subject. Typically, a normal control value is expressed as a range ("between X and Y"), a high threshold ("not higher than X"), or a low threshold ("not lower than X"). A subject whose measured value for a particular biomarker is within the normal control value is often referred to as being "within normal limits" for that biomarker. A normal reference standard or level can be a value or number derived from a normal subject without a disease or disorder (e.g., cancer); a subject who has been treated with a compound of the 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 levels of purified protein or RNA (e.g., any of those described herein) within the normal reference range can also be used as a reference.

[0781] The term "subject" as used herein refers to any organism to which a composition according to the invention can be administered, e.g., 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 seeking or in need of treatment, requiring treatment, being treated, will be treated in the future, or is a person or animal under the care of a trained professional for a particular disease or condition.

[0782] As used herein, the terms "treat," "treated," or "treating" refer to therapeutic treatment or any measure thereof aimed at alleviating (relieving) an undesirable physiological condition, disorder, or disease, or obtaining a beneficial or desired clinical result. Beneficial or desired clinical results include, but are not limited to: alleviation of symptoms; reduction in the extent of the condition, disorder, or disease; a stable (i.e., non-worsening) state of the condition, disorder, or disease; a delay or slowing of the onset of the progression of the condition, disorder, or disease; an improvement or remission (whether partial or complete) of the condition, disorder, or disease state; an improvement in at least one measurable physical parameter, not necessarily discernible by the patient; or an improvement or improvement of the condition, disorder, or disease. Treatment includes inducing a clinically significant response without undue side effects. Treatment also includes prolonging survival compared to expected survival if not receiving treatment. The compounds of the invention may also be used for "prophylactic treatment" or "prevention" of disorders, for example, in subjects at increased risk of developing the disorder.

[0783] The details of one or more embodiments of the invention are set forth in the description that follows. Other features, objects, and advantages of the invention will be apparent from the description and from the claims.

[0784] Details

[0785] The present disclosure features compounds that can be used to inhibit BRG1 and optionally BRM. These compounds can be used to modulate the activity of the BAF complex, for example, for treating BAF-related disorders such as cancer (e.g., BRG1 loss of function disorder). Exemplary compounds described herein include compounds having a structure according to Formula I or II, or a pharmaceutically acceptable salt thereof:

[0786]

[0787] in

[0788] Ring system A is a 5- to 9-membered heterocyclyl or heteroaryl;

[0789] m is 0, 1, 2, or 3;

[0790] k is 0, 1, or 2;

[0791] Each R 1 are independently halogen, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 Heteroalkyl, optionally substituted C 2 -C 9 Heterocyclic group, optionally substituted C 3 -C 8 Cycloalkyl or optionally substituted CH 2-C 3 -C 8 Cycloalkyl;

[0792] each X is independently halogen;

[0793] L is a linker; and

[0794] B is the degradation part.

[0795] Exemplary compounds described herein include compounds having a structure according to Formula I or II, or a pharmaceutically acceptable salt thereof:

[0796]

[0797] in

[0798] Ring system A is a 5- to 9-membered heterocyclyl or heteroaryl;

[0799] m is 0, 1, 2, or 3;

[0800] k is 0, 1, or 2;

[0801] L is the connector;

[0802] B is the degradation part;

[0803] Each R 1 are independently halogen, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 Heteroalkyl, optionally substituted C 3 -C 8 Cycloalkyl or optionally substituted C 2 -C 9 Heterocyclic group;

[0804] and

[0805] Each X is independently halogen.

[0806] Exemplary compounds described herein include compounds having a structure according to Formula I or II, or a pharmaceutically acceptable salt thereof:

[0807]

[0808] in

[0809] Ring system A is a 5- to 9-membered heterocyclyl or heteroaryl;

[0810] m is 0, 1, 2, or 3;

[0811] k is 0, 1, or 2;

[0812] L is a linker of formula III:

[0813] A 1 -(B 1 ) f -(C 1 ) g -(B 2 ) h -(B 3 ) i -(C 2 ) j -(B 4 ) k –A 2 ,

[0814] Formula III

[0815] in

[0816] A 1 is the bond between the linker and the ring system A;

[0817] A 2 is the bond between the degradable moiety and the linker;

[0818] B 1 , B 2 , B 3 and B 4 are each independently an optionally substituted ethynyl, an optionally substituted C 6 -C 10 Aryl, optionally substituted C 3 -C 10 Cycloalkyl, optionally substituted C 2 -C 9 Heterocyclic group, optionally substituted C 2 -C 9 Heteroaryl, O, S, S(O) 2 or NR N ;

[0819] Each R N are independently H, optionally substituted C 1–4 Alkyl, optionally substituted C 2–4 Alkenyl, optionally substituted C 2–4 Alkynyl, optionally substituted C 2–6 Heterocyclic group, optionally substituted C 6–12 Aryl or optionally substituted C 1–7 heteroalkyl;

[0820] C 1 and C 2 are each independently carbonyl, thiocarbonyl, sulfonyl or phosphoryl; and

[0821] f, g, h, i, j and k are each independently 0 or 1;

[0822] B is the degradation part;

[0823] Each R 1 are independently halogen, optionally substituted C 1 -C 6 Alkyl, optionally substituted C 1 -C 6 Heteroalkyl, optionally substituted C 3 -C 8 Cycloalkyl or optionally substituted C 2 -C 9 Heterocyclic group;

[0824] and

[0825] Each X is independently halogen.

[0826] In some embodiments, the compound has the structure of any one of Compounds 1-103 in Table 1, or a pharmaceutically acceptable salt thereof.

[0827] Other embodiments are described herein, as well as exemplary methods for synthesizing or producing these compounds.

[0828] Drug Use

[0829] The compounds described herein can be used in the methods of the invention and, while not being bound by theory, are believed to exert their ability to modulate the level, state and / or activity of the BAF complex by inhibiting the activity of BRG1 and / or BRM proteins within the BAF complex in mammals. BAF complex-associated disorders include, but are not limited to, disorders associated with BRG1 loss-of-function mutations.

[0830] One aspect of the invention relates to a method of treating a disorder associated with a BRG1 loss-of-function mutation, such as cancer (e.g., non-small cell lung cancer, colorectal cancer, bladder cancer, cancer of unknown primary site, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, or penile cancer) in a subject in need thereof. In some embodiments, the compound is administered in an amount and for a time effective to result 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) improved subject survival, (i) increased subject progression-free survival.

[0831] Treating cancer can result in a decrease in the size or volume of a tumor. For example, after treatment, the size of a tumor decreases by 5% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more) relative to its size before treatment. The size of a tumor can be measured by any reproducible measurement means. For example, the size of a tumor can be measured as a tumor diameter.

[0832] Treating cancer may further result in a reduction in the number of tumors. For example, after treatment, the number of tumors is 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 reproducible measurement means, for example, the number of tumors can be measured by counting tumors visible to the naked eye or visible under a specified magnification (e.g., 2x, 3x, 4x, 5x, 10x, or 50x).

[0833] Treatment of cancer can result in a reduction in the number of metastatic nodules in other tissues or organs away from the primary tumor site. For example, after treatment, the number of metastatic nodules is 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 measurement means. For example, the number of metastatic nodules can be measured by counting metastatic nodules visible to the naked eye or visible at a specified magnification (e.g., 2x, 10x, or 50x).

[0834] Compared to an untreated subject population, treating cancer can result in an increase in the average survival time of a subject population treated according to the present invention. For example, the average survival time increases by more than 30 days (more than 60 days, 90 days, or 120 days). The increase in the average survival time of a population can be measured by any reproducible means. For example, the increase in the average survival time of a population can be measured by calculating the average survival time after the group begins to be treated with the compound of the present invention. For example, the increase in the average survival time of a population can also be measured by calculating the average survival time after the group completes the first round of treatment with a pharmaceutically acceptable salt of the present invention.

[0835] Treatment of cancer can also result in a reduction in mortality in the treated subject population compared to an untreated population. For example, the mortality rate is reduced by more than 2% (e.g., more than 5%, 10%, or 25%). The reduction in mortality in the treated subject population can be measured by any reproducible means, such as by calculating the average number of disease-related deaths per unit time after the population begins treatment with a pharmaceutically acceptable salt of the present invention. For example, the reduction in population mortality can also be measured by calculating the average number of disease-related deaths per unit time after the first round of treatment with a pharmaceutically acceptable salt of the present invention.

[0836] Exemplary cancers that may be treated by the present 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-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 intestine cancer, and penile cancer.

[0837] Combination preparations and their uses

[0838] The compounds of the invention may be combined with one or more therapeutic agents. In particular, the therapeutic agent may be a therapeutic agent for the treatment or prophylactic treatment of any cancer described herein.

[0839] Combination therapy

[0840] The compounds of the present invention can be used alone, or in combination with additional therapeutic agents, such as other agents for treating cancer or symptoms associated therewith, or in combination with other treatment types for treating cancer. In combined therapy, the dosage of one or more therapeutic compounds can be reduced compared to the standard dosage when applied alone. For example, the dosage can be determined empirically by drug combination and arrangement, or can be inferred by isobolographic analysis (e.g., Black et al., Neurology 65: S3-S6, 2005). In this case, the dosage of these compounds should provide a therapeutic effect when combined.

[0841] In some embodiments, 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, epipodopyyllotoxins, antibiotics, L-asparaginase, topoisomerase inhibitors, interferons, platinum coordination complexes, anthracenedione-substituted ureas, methylhydrazine derivatives, adrenocortical inhibitors, adrenocortical steroids, progestins, estrogens, antiestrogens, androgens, antiandrogens, and gonadotropin-releasing hormone analogs. Also included are 5-fluorouracil (5-FU), folinic acid (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, carboquinone, meturedopa and uredopa; ethyleneimines and methylamelamines, including hexamethylmelamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethylolmelamine; acetogenins (particularly bullatacin and bullatacinone); camptothecins (including the synthetic analogue topotecan); bryostatin; callystatin; CC-1065 (including its synthetic analogues adolesin, carzeresin and biszeresin); cryptophycins (particularly cryptophycin 1 and cryptophycin 8); 8)); dolastatin; duocarmycin (including synthetic analogs KW-2189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil, naphthyl mustard, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard;Nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine and ranimnustine; antibiotics such as enediyne antibiotics (e.g., calicheamicins, especially calicheamicin gamma II and calicheamicin omega II (see, e.g., Agnew, Chem. Intl. Ed Engl. 33:183-186 (1994)); dynemicins, including dynemicin A; bisphosphonates such as clodronate; escitalopram, e.g., clodronate; esperamicin; and the new carcinogen chromophores and related chromoprotein enediyne antibiotic chromophores), aclacinomycins, actinomycins, authramycins, azaserine, bleomycin, actinomycin C, carabicin, caminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detoxibacin, 6-diazo-5-oxo-L-norleucine,; (doxorubicin, including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, mexilomycin, mitomycins such as mitomycin C, mycophenolic acid, nogamycin, olive mycin, peplomycin, porfibrinocin, puromycin, triferon-doxorubicin, rhodorubicin, streptomycin, streptozotocin, tuberculin, ubenimex, zorubicin, antimetabolites such as mefenamicin, fenvalerate, doxycycline, oxazolidinone ... Aminopterin and 5-fluorouracil (5-FU); folic acid analogs such as dimethoate, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiopurine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as caprotestosterone, drostanolone propionate, cyclothiocarb, melastosane, testolactone; anti-adrenal adenosines such as aminoglutethimide, mitotane, and trilostane; folic acid supplements such as folinic acid; aceglucuronosone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; colcemid; diacronil; eflomicin thine); elliptinium acetate; epothilone; etoglu; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansines such as maytansine and ansamitocin; mitoguanidine; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllic acid; 2-ethylhydrazide; methylbenzylhydrazine; Polysaccharide complex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizobactin; sizofuran; spirogermanamine; tricholomanic acid; triazoquinone; 2,2',2"-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, baculocin A, and anguidine); urethan; vindesine; dacarbazine; mannomustine; dibromomannitol; dibromodulanol; pipobroman; gacytosine; cytarabine ("Ara-C"); cyclophosphamide; thiotepa; taxanes, such as Paclitaxel (Bristol-Myers Squibb Oncology, Princeton, NJ), Cremophor-free, albumin-engineered paclitaxel nanoparticle formulation (American Pharmaceutical Partners, Schaumberg, Ill.) and doxetaxel (Rhone-Poulenc Rorer, Antony, France); chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum coordination complexes such as cisplatin, oxaliplatin, and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; Vinorelbine; novantrone; teniposide; edatrexate; daunomycin; aminopterin; xeloda; ibandronate; irinotecan (e.g., CPT-11); topoisomerase inhibitor RFS2000; difluoromethylornithine (DMFO); retinoic acid such as retinoic acid; capecitabine; and pharmaceutically acceptable salts, acids, or derivatives of any of the above. Two or more chemotherapeutic agents can be used in a mixture for administration in combination with the first therapeutic agent described herein. Suitable dosing regimens for combined chemotherapy are known in the art and are described in, for example, Saltz et al., (1999) Proc ASCO 18:233a and Douillard et al., (2000) Lancet 355:1041-7.

[0842] In some embodiments, the second therapeutic agent is a therapeutic agent that is a biologic, such as a cytokine used in cancer treatment (e.g., an interferon or an interleukin (e.g., IL-2)). In some embodiments, the biologic is an anti-angiogenic agent, such as an anti-VEGF agent, for example, bevacizumab In some embodiments, the biologic is an immunoglobulin-based biologic, such as a monoclonal antibody (e.g., a humanized antibody, a fully human antibody, an Fc fusion protein, or a functional fragment thereof) that agonizes a target to stimulate an anti-cancer response or antagonizes an antigen important for cancer. Such agents include Rituxan (rituximab); Zenapax (daclizumab); Sule (basiliximab); Synagis (palivizumab); Remicade (infliximab); Herceptin (trastuzumab); Mylotarg (gemtuzumab ozogamicin); Campath (alemtuzumab); Zevalin (ibritumomab tiuxetan); Humira (adalimumab); Xolair (omalizumab); Bexxar (tositumomab-I-131); Raptiva (efalizumab); Erbitux (cetuximab); Avastin (bevacizumab); Tysabri (natalizumab); Actemra (tocilizumab); Vectis (episib); Ibixi (panitumumab); Lucentis (ranibizumab); Soliris (eculizumab); Cimicifuga (becelimumab); Simponi (golimumab); Ilaris (canakinumab); Sedano (ustekinumab); Arzerra (ofatumumab); Proli (denosumab); Numax (motavizumab); ABThrax (ranibizumab); Belimumab; Yervoy (ipilimumab); Ansari (brentuximab); Perjeta (pertuzumab); Kadcyla (enmetuzumab); and Gazyva (otuzumab). Antibody-drug conjugates are also included.

[0843] The second agent can be a non-drug therapeutic agent. For example, the second therapeutic agent is radiation therapy, cryotherapy, hyperthermia and / or surgical removal of tumor tissue.

[0844] 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 an inhibitor (e.g., an inhibitory antibody or a small molecule inhibitor) of CTLA-4 (e.g., an anti-CTLA4 antibody such as ipilimumab / Yervoy or tremelimumab). In some embodiments, the checkpoint inhibitor is an inhibitor (e.g., an inhibitory antibody or a small molecule inhibitor) of PD-1 (e.g., nivolumab / Pembrolizumab / 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., an inhibitory antibody or Fc fusion or a small molecule inhibitor) (e.g., PDL2 / Ig fusion protein such as AMP 224). 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).

[0845] In any combination embodiments described herein, the first and second therapeutic agents are administered simultaneously or sequentially in either order. The first therapeutic agent can be administered immediately, up to 1 hour, up to 2 hours, up to 3 hours, up to 4 hours, up to 5 hours, up to 6 hours, up to 7 hours, up to 8 hours, up to 9 hours, up to 10 hours, up to 11 hours, up to 12 hours, up to 13 hours, 14 hours, up to 16 hours, up to 17 hours, up to 18 hours, up to 19 hours, up to 20 hours, up to 21 hours, up to 22 hours, up to 23 hours, up to 24 hours, or up to 1-7 days, 1-14 days, 1-21 days, or 1-30 days before or after the second therapeutic agent.

[0846] Pharmaceutical composition

[0847] 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. Thus, in one aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention admixed with a suitable diluent, carrier or excipient.

[0848] The compound of the present invention can be used in the form of free alkali, in the form of salt, solvate and as a prodrug. All forms are within the scope of the present invention. As will be appreciated by those skilled in the art, according to the method of the present invention, the described compound or its salt, solvate or prodrug can be applied to the patient in various forms according to the selected route of administration. The compound of the present invention can be applied, for example, by oral, parenteral, buccal, sublingual, nasal, rectal, patch, pump or transdermal administration and a pharmaceutical composition prepared accordingly. Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, epithelial, nasal, intrapulmonary, intrathecal, rectal and topical administration modes. Parenteral administration can be carried out by continuous infusion in the selected time period.

[0849] The compounds of the present invention can be administered orally, for example with an inert diluent or with an assimilable edible carrier, or they can be encapsulated in a hard or soft shell gelatin capsule, or they can be compressed into tablets, or they can be directly incorporated into the food in the diet. For oral treatment administration, the compounds of the present invention can be combined with excipients and used in the form of ingestible tablets, buccal tablets, lozenges, capsules, elixirs, suspensions, syrups and wafers. The compounds of the present invention can also be administered parenterally. Solutions of the compounds of the present invention can be prepared in water suitably mixed with a surfactant (such as hydroxypropylcellulose). Dispersions can also be prepared in glycerol, liquid polyethylene glycol, DMSO and mixtures thereof containing or not containing alcohol and in oil. Under normal storage and use conditions, these preparations can contain preservatives to prevent the growth of microorganisms. Conventional procedures and ingredients for selecting and preparing suitable preparations are described in, for example, Remington's Pharmaceutical Sciences (2003, 20th edition) and The United States Pharmacopeia: The National Formulary (USP 24NF19) published in 1999. The drug form suitable for injection includes sterile aqueous solutions or dispersions, and sterile powders for instant preparation of sterile injection solutions or dispersions. In all cases, the form must be sterile and must be a fluid that can be easily applied via a syringe. Compositions for nasal administration can be conveniently formulated as aerosols, drops, gels, and powders. Aerosol preparations typically include solutions or fine suspensions of active substances in physiologically acceptable aqueous or non-aqueous solvents, and are typically present in a sealed container in a sterile form in a single dose or multiple doses, and the container can be in the form of a cartridge or refill to be used with a nebulizer. Alternatively, the sealed container can be a unit dispensing device, such as a single-dose nasal inhaler or an aerosol dispenser equipped with a metering valve, which is intended to be discarded after use. In the case where the dosage form includes an aerosol dispenser, it contains a propellant, which can be a compressed gas, such as compressed air, or an organic propellant, such as a fluorochlorocarbon. Aerosol dosage forms can also take the form of a pump-atomizer. Compositions suitable for buccal or sublingual administration include tablets, lozenges and pastilles, wherein the active ingredient is formulated with a carrier, such as sugar, gum arabic, tragacanth, gelatin and glycerol. Compositions for rectal administration are conveniently in the form of suppositories containing conventional suppository bases, such as cocoa butter. The compounds described herein can be administered intratumorally, for example as intratumoral injections. Intratumoral injections are injected directly into the tumor vasculature, and are particularly contemplated for discrete, solid, palpable tumors. Local, regional or systemic administration may also be appropriate.The compounds described herein may advantageously be contacted by administering a single injection or multiple injections (e.g., spaced approximately 1 cm apart) to the tumor. In the case of surgical intervention, the present invention may be used prior to surgery, such as to allow resection of an inoperable tumor. Continuous administration may also be employed where appropriate, for example, by implanting a catheter into the tumor or tumor vasculature.

[0850] The compounds of the invention can be administered to animals, such as humans, alone or in combination with a pharmaceutically acceptable carrier as described herein, the proportions of which will depend on the solubility and chemical properties of the compound, the chosen route of administration, and standard pharmaceutical practice.

[0851] dose

[0852] The dosage of the compound of the present invention and / or the composition comprising the compound of the present invention can vary with many factors, such as the pharmacodynamic properties of the compound; the mode of administration; the age, health and weight of the recipient; the nature and extent of the symptoms; the frequency of treatment and the type of concurrent treatment (if any); and the clearance rate of the compound in the animal to be treated. Those skilled in the art can determine the appropriate dosage based on the above factors. The compound of the present invention can be initially administered at a suitable dose, which can be adjusted as needed according to the clinical response. In general, when the compound of the present invention is administered to a person at a daily dose of, for example, 0.05 mg to 3000 mg (measured in solid form), satisfactory results can be obtained. The dosage range includes, for example, between 10-1000 mg (e.g., 50-800 mg). In some embodiments, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or 1000 mg of the compound is administered.

[0853] Alternatively, the patient's weight can be used to calculate the dosage. For example, the dosage of the compound or its pharmaceutical composition administered to the patient can be 0.1-100 mg / kg (e.g., 0.25-25 mg / kg). In an exemplary, non-limiting embodiment, the dosage can be 0.5-5.0 mg / kg (e.g., 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, or 5.0 mg / kg) or 5.0-20 mg / kg (e.g., 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg / kg). Example

[0854] The following abbreviations are used throughout the following examples.

[0855] Ac Acetyl

[0856] ACN or MeCN Acetonitrile

[0857] AcOH Acetic acid

[0858] Ac 2 O Acetic anhydride

[0859] aq. water-based

[0860] Boc tert-Butyloxycarbonyl

[0861] Bu or n-Bu Butyl

[0862] CDI 1,1'-Carbonyldiimidazole

[0863] DCE or 1,2-DCE 1,2-dichloroethane

[0864] DCM Dichloromethane

[0865] DIAD Diisopropyl azodicarboxylate

[0866] DIPEA or DIEA NN-diisopropylethylamine

[0867] DMAP 4-(dimethylamino)pyridine

[0868] DME 1,2-Dimethoxyethane

[0869] DMF NN-Dimethylformamide

[0870] DMSO Dimethyl sulfoxide

[0871] EA or EtOAc Ethyl acetate

[0872] EDCI N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride

[0873] equiv equivalent

[0874] E 3 N or TEA Triethylamine

[0875] EtOH

[0876] FA Formic acid

[0877] h or hr hour

[0878] HATU 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo

[0879] [4,5-b]Pyridinium 3-oxide hexafluorophosphate HOAt 1-Hydroxy-7-azabenzotriazole

[0880] HOBt or HOBT 1-Hydroxybenzotriazole hydrate

[0881] iPr Isopropyl

[0882] MeOH Methanol

[0883] Me 4 t-BuXphos Di-tert-butyl-[2,3,4,5-tetramethyl-6-(2,4,6-triisopropylphenyl)benzene phosphane

[0884] min

[0885] MTBE tert-butyl methyl ether

[0886] n-BuLi n-butyllithium

[0887] NMP 1-Methyl-2-pyrrolidone

[0888] OAc acetate

[0889] Pd / C Palladium on Carbon

[0890] PDC Pyridinium dichromate

[0891] PdCl 2 (dtbpf) or dichloro[1,1'-bis(di-tert-butylphosphino)ferrocene]palladium(II) Pd(dtbpf)Cl 2

[0892] PdCl 2 (dppf) or [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) Pd(dppf)Cl 2

[0893] Pd 2 (dba) 3 Tris(dibenzylideneacetone)dipalladium(0)

[0894] Pd(PPh 3 ) 4 Tetrakis(triphenylphosphine)palladium(0)

[0895] Pd(PPh 3 ) 2 Cl 2 Dichlorobis(triphenylphosphine)palladium(II)

[0896] PE Petroleum Ether

[0897] PPh 3Triphenylphosphine

[0898] Pr n-propyl

[0899] Py Pyridine

[0900] rac racemic

[0901] Rf retention factor

[0902] rt or rt room temperature

[0903] sat. saturated

[0904] SFC Supercritical Fluid Chromatography

[0905] t-Bu tert-butyl

[0906] tBuXphos-Pd-G3 or [2-(2-aminophenyl)phenyl]-methylsulfonyloxypalladium; ditert- tBuXphos Pd G 3 or butyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphane t-BuXphos-Pd(gen 3)

[0907] TFA Trifluoroacetic acid

[0908] Tf 2 O Trifluoromethanesulfonic anhydride

[0909] THF Tetrahydrofuran

[0910] TLC Thin layer chromatography

[0911] Xantphos-Pd-G3 [2-(2-aminophenyl)phenyl]-methylsulfonyloxy-palladium; (5-

[0912] Diphenylphosphanyl-9,9-dimethyl-xanthene-4-yl)-diphenyl

[0913] -Phosphine

[0914] Example 1. Preparation of compounds

[0915] Preparation of methyl 2-(3-hydroxy-1,2-oxazol-5-yl)-3-methylbutanoate (I-1)

[0916] Step 1: Preparation of 2-(3-bromoisoxazol-5-yl)ethan-1-ol

[0917]

[0918] A solution of 3-butyn-1-ol (552.89 g, 7888.26 mmol, 4 equivalents) and KHCO3 (592.30 g, 5916.197 mmol, 3 equivalents) in EtOAc (2600 mL) and H2O (260 mL) was stirred at room temperature. 1-bromo-N-hydroxymethylimino bromide (400.00 g in EA (840 mL), 1972.066 mmol, 1.00 equivalent) was added dropwise to the above mixture over 60 minutes at room temperature. The resulting mixture was stirred at room temperature overnight. The reaction mixture was washed with water (500 mL x 2), and the combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with PE / EtOAc (30: 1) to provide an intermediate title compound (338.2 g, 88.98%) as an off-white solid. LCMS (ESI) m / z [M+H] + = 192.

[0919] Step 2: Preparation of 2-(3-bromoisoxazol-5-yl)acetic acid

[0920]

[0921] A solution of 2-(3-bromoisoxazol-5-yl)ethan-1-ol (360.00 g) in acetone (3600 mL) was stirred at 0°C under a nitrogen atmosphere. Jones reagent (1760 mL) was added dropwise to the above mixture at 0°C over 1 hour. The resulting mixture was stirred at room temperature overnight. The reaction was quenched with water / ice at 0°C. The resulting mixture was extracted with EtOAc (1000 mL x 3). The combined organic layers were washed with water (500 mL x 2) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to provide the title compound (348.6 g, crude) as a green solid, which was used directly without further purification. (LCMS (ESI) m / z[M+H]+=206.

[0922] Step 3: Preparation of ethyl 2-(3-bromoisoxazol-5-yl)acetate

[0923]

[0924] A solution of 2-(3-bromoisoxazol-5-yl)acetic acid (397.6 g, 1930.144 mmol, 1.00 equiv) and H2SO4 (18.92 g, 193.014 mmol, 0.1 equiv) in EtOH (2000 mL) was stirred at 70 ° C for 2 hours. The reaction mixture was concentrated under reduced pressure. The residue was diluted with EtOAc (3000 mL), washed with water (500 mL x 2), and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with PE / EtOAc (35: 1) to provide the title compound (355 g, 78.61%) as a colorless oil. (LCMS (ESI) m / z [M+H] + = 234.

[0925] Step 4: Preparation of ethyl 2-(3-bromoisoxazol-5-yl)-3-methylbutanoate

[0926]

[0927] To a stirred solution of t-BuOK (244.51 g, 2179.031 mmol, 1.5 equiv) and ethyl 2-(3-bromoisoxazole-5-yl)acetate (340.00 g, 1452.687 mmol, 1.00 equiv) in THF (2000 mL) was added 2-iodopropane (321.03 g, 1888.493 mmol, 1.3 equiv) dropwise under a nitrogen atmosphere at 0 ° C. The resulting mixture was stirred at room temperature overnight and then diluted with water / ice at 0 ° C. The resulting mixture was extracted with EtOAc (1000 mL x 2). The combined organic layer was washed with water (500 mL x 1) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / THF (10:1) to provide the title compound (284.1 g, 70.82%) as a colorless oil. (LCMS (ESI) m / z [M+H] + = 276.

[0928] Step 5: Preparation of 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid

[0929]

[0930] To a stirred solution of ethyl 2-(3-bromoisoxazol-5-yl)-3-methylbutanoate (90.00 g, 325.933 mmol, 1.00 equiv) in MeOH (270 mL) at 0 ° C. was added a solution of KOH (274.30 g, 4888.995 mmol, 15.00 equiv) in MeOH (210 mL). The reaction mixture was stirred at 80 ° C overnight. The resulting solution was acidified to pH 4 with 1M HCl solution (aqueous solution) and concentrated under reduced pressure. The resulting mixture was diluted with EtOAc (1800 mL) and filtered. The filter cake was washed with EtOAc (100 mLx3). The filtrate was concentrated under reduced pressure to provide the title compound (62.9 g, 96.88%) as a yellow oil, which was used directly without further purification. LCMS (ESI) m / z: [M+H] + = 200.

[0931] Step 6: Preparation of 2-(3-hydroxy-1,2-oxazol-5-yl)-3-methylbutanoic acid

[0932]

[0933] To a stirred solution of 2-(3-methoxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (62.90 g, 315.754 mmol, 1.00 equiv) in HOAc (450.00 mL) was added 48% HBr (450.00 mL) at room temperature. The resulting mixture was stirred at 60 °C for 16 hours. The resulting mixture was concentrated under reduced pressure and the residue was purified by flash C18-flash chromatography with an elution gradient of 0 to 100% MeCN / water (containing 0.05% FA). The pure fractions were evaporated to dryness to afford the title compound (43.3 g, 74.05%) as a white solid. LCMS (ESI) m / z: [M+H]+=186.

[0934] Step 7: Preparation of methyl 2-(3-hydroxy-1,2-oxazol-5-yl)-3-methylbutanoate (I-1)

[0935]

[0936] To a stirred solution of 2-(3-hydroxy-1,2-oxazol-5-yl)-3-methylbutanoic acid (20 g, 108.004 mmol, 1.00 equiv) in MeOH (72 mL) was added SOCl2 (35.26 mL, 486.059 mmol, 4.50 equiv) at 0 ° C. The resulting mixture was stirred at room temperature for 16 hours. The resulting mixture was concentrated under reduced pressure, and the residue was diluted with water (30 mL) and extracted with EtOAc (50 mL x 3). The combined organic layer was washed with saturated NaCl aqueous solution (30 mL), dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography with an elution gradient of 0 to 100% THF / petroleum ether. The pure fraction was evaporated to dryness to provide compound I-1 (15.1 g, 70.18%) as an off-white solid. 1HNMR (400MHz, DMSO-d6) δ11.24 (s, 1H), 5.95 (s, 1H), 3.71–3.58 (m, 4H), 2.32–2.20 (m, 1H), 0.88 (dd, J = 34.2, 6.7Hz, 6H). LCMS (ESI) m / z: [M+H]+ = 200.

[0937] Preparation of methyl 2-(3-((2-chloropyrimidin-4-yl)oxy)isoxazol-5-yl)-3-methylbutanoate (Intermediate 2)

[0938]

[0939] A solution of methyl 2-(3-hydroxy-1,2-oxazol-5-yl)-3-methylbutanoate (200 mg, 1.004 mmol, 1 eq.) and 4-bromo-2-chloropyrimidine (233.04 mg, 1.205 mmol, 1.2 eq.), Cs2CO3 (981.9 mg, 3.012 mmol, 3.0 eq.) in DMF (5 mL) was stirred at 100 °C for 1 hour under nitrogen atmosphere. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.1% FA), 10% to 100% gradient in 20 minutes; detector, UV 254 nm. The mixture was concentrated to provide intermediate 2 (258 mg, crude). LCMS (ESI) m / z: [M+H]+=312

[0940] Preparation of methyl 2-[3-(2-chloropyrimidin-5-yl)-1,2-oxazol-5-yl]-3-methylbutanoate (I-2)

[0941]

[0942] Step 1: Preparation of (E)-N-[(2-chloropyrimidin-5-yl)methylidene]hydroxylamine

[0943]

[0944] To a stirred solution of 2-chloropyrimidine-5-carboxaldehyde (5 g, 35.078 mmol, 1 eq.) and NH2OH.HCl (4.93 g, 70.945 mmol, 2.02 eq.) in EtOH (250 mL) was added NaOAc (14.48 g, 176.512 mmol, 5.03 eq.) at room temperature. The resulting mixture was stirred at room temperature for 2 hours. The solvent was removed under reduced pressure. The residue was dissolved in EtOAc (500 mL), washed with brine (500 mL), and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to provide (E)-N-[(2-chloropyrimidin-5-yl)methylidene]hydroxylamine (4.6 g, crude product) as a light yellow solid, which was used directly in the next step without further purification. LCMS (ESI) m / z: [M+H]+=158.

[0945] Step 2: Preparation of (Z)-2-chloro-N-hydroxypyrimidine-5-carbonimidoyl chloride

[0946]

[0947] A solution of (E)-N-[(2-chloropyrimidin-5-yl)methylidene]hydroxylamine (4.6 g, 29.195 mmol, 1 eq.) and NCS (4.4 g, 32.951 mmol, 1.13 eq.) in DMF (150 mL) was stirred at room temperature for 2 hours. The mixture was diluted with EtOAc (500 mL). The resulting mixture was washed with water (3 x 300 mL), brine (1 x 300 mL), and the organic phase was dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to provide (Z)-2-chloro-N-hydroxypyrimidine-5-carboximidoyl chloride (4.8 g, crude product) as a yellow solid. LCMS (ESI) m / z: [M+H]+=192.

[0948] Step 3: Preparation of methyl 2-[3-(2-chloropyrimidin-5-yl)-1,2-oxazol-5-yl]acetate

[0949]

[0950] A solution of (Z)-2-chloro-N-hydroxypyrimidine-5-carboximidoyl chloride (4.8 g, 25.00 mmol, 1 eq.) in EtOAc (80 mL) was treated with NaHCO (3 g, 35.712 mmol, 1.43 eq.) at 0 ° C under a dry nitrogen atmosphere for 30 minutes, and then methyl butane-3-ynoate (2.02 g, 20.591 mmol, 0.82 eq.) was added in portions at 0 ° C. The resulting mixture was stirred at room temperature for 12 hours. The resulting mixture was diluted with water (150 mL) and extracted with EtOAc (2x400 mL). The combined organic layer was washed with brine (1x 400 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (3:1) to provide methyl 2-[3-(2-chloropyrimidin-5-yl)-1,2-oxazol-5-yl]acetate (2.5 g, 38.64%) as a light yellow solid. LCMS (ESI) m / z: [M+H]+=254.

[0951] Step 4: Preparation of [3-(2-methoxypyrimidin-5-yl)-1,2-oxazol-5-yl]acetic acid

[0952]

[0953] A solution of 2-[3-(2-chloropyrimidin-5-yl)-1,2-oxazol-5-yl]methyl acetate (3g, 11.828mmol, 1 eq) and NaOMe (1.92g, 35.484mmol, 3.00 eq) in MeOH (50mL) was stirred at room temperature for 1 hour under a dry nitrogen atmosphere. The mixture was acidified to pH 6 with HCl (aqueous solution). The residue was dissolved in EtOAc (300mL). The resulting mixture was washed with water (2x 300mL). The combined organic layer was washed with brine (1x 300mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to provide [3-(2-methoxypyrimidin-5-yl)-1,2-oxazol-5-yl]acetic acid (2.5g, crude product) as a light yellow solid, which was directly used in the next step without further purification. LCMS (ESI) m / z: [M+H]+=236.

[0954] Step 5: Preparation of methyl 2-[3-(2-methoxypyrimidin-5-yl)-1,2-oxazol-5-yl]acetate

[0955]

[0956] A solution of [3-(2-methoxypyrimidin-5-yl)-1,2-oxazol-5-yl]acetic acid (2.4 g, 10.204 mmol, 1 eq.) and (trimethylsilyl)diazomethane (2.33 g, 20.408 mmol, 2 eq.) in DCM (20 mL) and MeOH (5 mL) was stirred at room temperature for 30 minutes. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with PE / EA (3:1) to provide methyl 2-[3-(2-methoxypyrimidin-5-yl)-1,2-oxazol-5-yl]acetate (1.2 g, 45.77%) as a white solid. LCMS (ESI) m / z: [M+H]+=250.

[0957] Step 6: Preparation of methyl 2-[3-(2-methoxypyrimidin-5-yl)-1,2-oxazol-5-yl]-3-methylbutanoate

[0958]

[0959] A solution of 2-[3-(2-methoxypyrimidine-5-yl)-1,2-oxazol-5-yl]methyl acetate (2.5 g, 10.031 mmol, 1 eq.) in THF (20 mL) was treated with t-BuOK (1.2 g, 10.694 mmol, 1.07 eq.) at 0 ° C for 30 minutes under a dry nitrogen atmosphere, and then 2-iodopropane (1.5 g, 8.824 mmol, 0.88 eq.) was added dropwise at 0 ° C. The resulting mixture was stirred at room temperature for 12 hours. The mixture was acidified to pH 6 with HCl (aqueous solution). The resulting mixture was extracted with EtOAc (2x200 mL). The combined organic layer was washed with brine (2x 200 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (3:1) to provide methyl 2-[3-(2-methoxypyrimidin-5-yl)-1,2-oxazol-5-yl]-3-methylbutanoate (310 mg, 10.08%) as a yellow oil. LCMS (ESI) m / z: [M+H]+=292.

[0960] Step 7: Preparation of methyl 2-[3-(2-chloropyrimidin-5-yl)-1,2-oxazol-5-yl]-3-methylbutanoate

[0961]

[0962] A solution of methyl 2-[3-(2-methoxypyrimidin-5-yl)-1,2-oxazol-5-yl]-3-methylbutanoate (200 mg, 0.687 mmol, 1 eq.) and POCl3 (1.9 mL, 20.61 mmol, 30 eq.) in DMF (1.5 mL) was stirred at 60 ° C for 3 hours under a dry nitrogen atmosphere. The residue was dissolved in EtOAc (100 mL). The resulting mixture was washed with brine (2×100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to provide methyl 2-[3-(2-chloropyrimidin-5-yl)-1,2-oxazol-5-yl]-3-methylbutanoate (160 mg, crude product) as a brown oil, which was used directly in the next step without further purification. LCMS (ESI) m / z: [M+H]+=296.

[0963] Preparation of tert-butyl (2S, 4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]-1-(2R)-3-methyl-2-[3-(piperidin-4-yl)-1,2-oxazol-5-yl]butyryl]pyrrolidine-2-carboxamide (I-3)

[0964]

[0965] Step 1: Preparation of tert-butyl 4-[(1E)-(hydroxyimino)methyl]piperidine-1-carboxylate (Intermediate 2)

[0966]

[0967] Hydroxylamine hydrochloride (1.95 g, 28.133 mmol, 1.2 equiv) and Na2CO3 (1.24 g, 11.722 mmol, 0.5 equiv) were added to a stirred solution of tert-butyl 4-formylpiperidine-1-carboxylate (5 g, 23.444 mmol, 1.00 equiv) in MeOH (10 mL) and H2O (10 mL) at 0°C. The resulting mixture was stirred at room temperature overnight. The desired product can be detected by LCMS. The resulting mixture was concentrated under reduced pressure. The resulting mixture was extracted with EtOAc (3x20 mL). The combined organic layers were washed with brine (50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to provide intermediate 2 (6 g, crude) as a colorless oil. LCMS (ESI) m / z: [M+H]+=229.

[0968] Step 2: Preparation of tert-butyl 4-[(1Z)-chloro(hydroxyimino)methyl]piperidine-1-carboxylate (Intermediate 3)

[0969]

[0970] A mixture of intermediate 2 and NCS (3.5 g, 26.282 mmol, 1.0 equiv) in DMF (20 mL) was stirred at room temperature for 2 hours. The desired product can be detected by LCMS. The resulting mixture was diluted with water (50.00 mL) and extracted with EtOAc (3x50 mL). The combined organic layer was washed with brine (50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to provide intermediate 3 (7.8 g, crude) as a colorless oil. LCMS (ESI) m / z[M+H]+=263.

[0971] Step 3: Preparation of tert-butyl 4-[5-(2-methoxy-2-oxoethyl)-1,2-oxazol-3-yl]piperidine-1-carboxylate (Intermediate 4)

[0972]

[0973] A mixture of intermediate 3 (7.8 g, crude) and NaHCO3 (3.8 g, 45.675 mmol, 1.5 eq) in EtOAc (100 mL) was stirred at room temperature for 30 minutes. To the above mixture was added but-3-ynoic acid methyl ester (2.99 g, 30.450 mmol, 1 eq) at 0°C. The resulting mixture was stirred at room temperature overnight. The desired product can be detected by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.05% FA), 0% to 100% gradient in 30 minutes; detector, UV254 nm. The resulting mixture was concentrated under reduced pressure to provide intermediate 4 (4.1 g, 41.51%) as a light yellow oil. LCMS (ESI) m / z: [M+H]+=325.

[0974] Step 4: Preparation of tert-butyl 4-[5-(1-methoxy-3-methyl-1-oxobutan-2-yl)-1,2-oxazol-3-yl]piperidine-1-carboxylate (Intermediate 5)

[0975]

[0976] To a mixture of intermediate 4 (1.0 g, 3.083 mmol, 1.5 eq.) and Na2SO4 (1.0 g) in THF (10 mL) was added t-BuOK (518.90 mg, 4.625 mmol, 1.5 eq.) and 2-iodopropane (628.87 mg, 3.700 mmol, 1.2 eq.) at 0°C under a nitrogen atmosphere. The resulting mixture was stirred at 0°C for 3 hours under a nitrogen atmosphere. The desired product can be detected by LCMS. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.05% FA), 0% to 100% gradient in 30 minutes; detector, UV 254 nm. The resulting mixture was concentrated under reduced pressure to provide intermediate 5 (330 mg, 29.21%) as a light yellow oil. LCMS (ESI) m / z: [M+H]+=367.

[0977] Step 5: Preparation of 2-{3-[1-(tert-butoxycarbonyl)piperidin-4-yl]-1,2-oxazol-5-yl}-3-methylbutanoic acid (Intermediate 6)

[0978]

[0979] To a stirred solution of intermediate 5 (320 mg, 0.873 mmol, 1.00 equiv) in MeOH (5 mL) was added dropwise LiOH (62.74 mg, 2.619 mmol, 3 equiv) in H2O (5 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 hours. The desired product can be detected by LCMS. The resulting mixture was concentrated under reduced pressure. Aqueous HCl solution (6 M) was added to the above mixture to adjust the pH to ~5. The resulting mixture was extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to provide intermediate 6 (316 mg crude) as an off-white solid. LCMS (ESI) m / z: [M+H]+=353.

[0980] Step 6: Preparation of tert-butyl 4-(5-{1-[(2S,4R)-4-hydroxy-2-{[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]carbamoyl}pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}-1,2-oxazol-3-yl)piperidine-1-carboxylate (Intermediate 7)

[0981]

[0982] A mixture of intermediate 6 (310 mg, 0.880 mmol, 1.00 eq) and HATU (668.90 mg, 1.760 mmol, 2 eq) in DMF (5 mL) was stirred at room temperature for 30 minutes. To the above mixture was added (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (291.53 mg, 0.880 mmol, 1 eq) at room temperature. The resulting mixture was stirred at room temperature for another 2 hours. The desired product can be detected by LCMS. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.05% FA), 0% to 100% gradient in 30 minutes; detector, UV 254 nm. The resulting mixture was concentrated under reduced pressure to provide Intermediate 7 (242 mg, 37.31%) as a light brown solid. LCMS (ESI) m / z: [M+H]+=666.

[0983] Step 7: Preparation of tert-butyl 4-{5-[(2R)-1-[(2S,4R)-4-hydroxy-2-{[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]carbamoyl}pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl]-1,2-oxazol-3-yl}piperidine-1-carboxylate (Intermediate 8)

[0984]

[0985] The product was purified by preparative SFC using the following conditions (column: CHIRALART Amylose-SA, 3*25 cm, 5 μm; mobile phase A: CO2, mobile phase B: MeOH--HPLC; flow rate: 50 mL / min; gradient: isocratic 45% B; column temperature (°C): 35; back pressure (bar): 100; wavelength: 205 nm; RT1 (min): 3.65; RT2 (min): 4.88; sample solvent: MeOH--HPLC; injection volume: 1 mL) to provide intermediate 8 (second peak) as a light brown solid (208.1 mg, 43.52%). LCMS (ESI) m / z: [M+H]+=666.

[0986] Step 8: Preparation of tert-butyl (2S, 4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]-1-[(2R)-3-methyl-2-[3-(piperidin-4-yl)-1,2-oxazol-5-yl]butyryl]pyrrolidine-2-carboxamide (Intermediate 9)

[0987]

[0988] To a stirred solution of intermediate 8 (200 mg, 0.300 mmol, 1.00 equiv) in DCM (2 mL) was added 1 M HCl / 1,4-dioxane (2 mL) dropwise at room temperature. The resulting mixture was stirred at room temperature for 1 hour. The desired product can be detected by LCMS. The resulting mixture was concentrated under reduced pressure to provide intermediate 9 (247.5 mg) as a light yellow solid. LCMS (ESI) m / z: [M+H]+=566.

[0989] Preparation of (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]-1-[(2S)-3-methyl-2-[3-(piperazin-1-yl)-1,2-oxazol-5-yl]butyryl]pyrrolidine-2-carboxamide (I-4) and (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]-1-[(2R)-3-methyl-2-[3-(piperazin-1-yl)-1,2-oxazol-5-yl]butyryl]pyrrolidine-2-carboxamide (I-5)

[0990]

[0991] Step 1: Preparation of methyl 3-methyl-2-[3-[(1,1,2,2,3,3,4,4,4-nonafluorobutanesulfonyl)oxy]-1,2-oxazol-5-yl]butanoate (Intermediate 2)

[0992]

[0993] To a stirred solution of 2-(3-hydroxy-1,2-oxazol-5-yl)-3-methylbutanoic acid methyl ester (100.00 mg, 0.502 mmol, 1.00 equiv) in MeCN (0.50 mL) was added perfluorobutanesulfonyl fluoride (303.29 mg, 1.004 mmol, 2.00 equiv) and K2CO3 (208.13 mg, 1.506 mmol, 3.00 equiv) at room temperature. The resulting mixture was stirred for 3 hours and then carefully quenched with water at 0 ° C. The resulting mixture was extracted with EA (2 x 50 mL), and the combined organic layers were washed with brine (50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography eluted with PE / EA (2 / 1) to provide intermediate 2 (217 mg, crude) as a white solid. LCMS (ESI) m / z: [M+H]+=482.

[0994] Step 2: Preparation of tert-butyl 4-[5-(1-methoxy-3-methyl-1-oxobutan-2-yl)-1,2-oxazol-3-yl]piperazine-1-carboxylate (Intermediate 3)

[0995]

[0996] To a stirred solution of intermediate 2 (217.00 mg, 0.451 mmol, 1.00 equiv) in DMF (3.00 mL) was added tert-butyl piperazine-1-carboxylate (83.98 mg, 0.451 mmol, 1.00 equiv) at room temperature. The resulting mixture was stirred at 130 °C for 1 hour. The mixture was allowed to cool to room temperature. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.1% FA), 0 to 100% gradient in 30 minutes. This provided intermediate 3 (54 mg, 32.59%) as a yellow oil. LCMS (ESI) m / z: [M+H]+=368.

[0997] Step 3: Preparation of 2-[3-[4-(tert-butoxycarbonyl)piperazin-1-yl]-1,2-oxazol-5-yl]-3-methylbutanoic acid (Intermediate 4)

[0998]

[0999] To a stirred solution of intermediate 3 (54.00 mg, 0.147 mmol, 1.00 equiv) in MeOH (0.80 mL) was added THF (0.80 mL) and H2O (0.80 mL) at room temperature, followed by LiOH.H2O (18.50 mg, 0.441 mmol, 3.00 equiv). The resulting mixture was stirred at room temperature for another 1 hour. The mixture was acidified to pH 6 with HCl (1 M, aqueous solution) and then extracted with EA (2 x 50 mL). The combined organic layers were washed with saturated brine (50 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure. This provided intermediate 4 (52 mg, crude) as a yellow solid. LCMS (ESI) m / z: [M+H]+=354.

[1000] Step 4: Preparation of tert-butyl 4-(5-[1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl]-1,2-oxazol-3-yl)piperazine-1-carboxylate (Intermediate 6)

[1001]

[1002] To a stirred solution of intermediate 4 (52.00 mg, 0.119 mmol, 1.00 equiv) in DMF (2.00 mL) was added HATU (135.56 mg, 0.357 mmol, 3.00 equiv) and DIEA (76.80 mg, 0.595 mmol, 5.00 equiv) at room temperature. To the above mixture was added (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (70.90 mg, 0.214 mmol, 1.80 equiv) at room temperature. The resulting mixture was stirred for another 1 hour. The mixture was directly purified by reverse phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.1% FA), 0 to 100% gradient in 30 minutes. This provided intermediate 6 (73 mg, 92.12%) as a white solid. LCMS (ESI) m / z: [M+H]+=667.

[1003] Step 5: Preparation of (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]-1-[(2R)-3-methyl-2-[3-(piperazin-1-yl)-1,2-oxazol-5-yl]butyryl]pyrrolidine-2-carboxamide (Intermediate 7); (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]-1-[(2S)-3-methyl-2-[3-(piperazin-1-yl)-1,2-oxazol-5-yl]butyryl]pyrrolidine-2-carboxamide (Intermediate 8)

[1004]

[1005] Intermediate 6 (73 mg) was purified by SFC under the following conditions: column, CHIRAL ART Amylose-C NEO, 3*25 cm, 5 um; mobile phase, MeOH. This provided intermediate 7 (37 mg, second peak). LCMS (ESI) m / z: [M+H]+=667, and intermediate 8 (34 mg, first peak). LCMS (ESI) m / z: [M+H]+=667.

[1006] Step 6: Preparation of (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]-1-[(2R)-3-methyl-2-[3-(piperazin-1-yl)-1,2-oxazol-5-yl]butyryl]pyrrolidine-2-carboxamide (I-4)

[1007]

[1008] To a stirred solution of intermediate 7 (37.00 mg, 0.055 mmol, 1.00 equiv) in DCM (1.50 mL) at 0°C was added HCl / 1,4-dioxane (1.50 mL, 26.276 mmol, 473.57 equiv). The resulting mixture was stirred at room temperature for 1 hour and then concentrated under reduced pressure. This provided I-4 (45 mg, crude) as a yellow oil. LCMS (ESI) m / z: [M+H]+=567.

[1009] Step 7: Preparation of (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]-1-[(2S)-3-methyl-2-[3-(piperazin-1-yl)-1,2-oxazol-5-yl]butyryl]pyrrolidine-2-carboxamide (I-5)

[1010]

[1011] To a stirred solution of intermediate 8 (34.00 mg, 0.051 mmol, 1.00 equiv) in DCM (1.50 mL) was added HCl / 1,4-dioxane (1.50 mL, 26.276 mmol, 515.35 equiv) at 0°C. The resulting mixture was stirred at room temperature for 1 hour and then concentrated under reduced pressure. This provided I-5 (45 mg, crude) as a yellow oil. LCMS (ESI) m / z: [M+H]+=567.

[1012] Preparation of 2-[6-(azetidin-3-yl)thieno[2,3-c]pyridazin-3-yl]phenol (I-6)

[1013]

[1014] Step 1: Preparation of tert-butyl 3-[2-(3,6-dichloropyridazin-4-yl)ethynyl]azetidine-1-carboxylate

[1015]

[1016] To a stirred mixture of 3,6-dichloro-4-iodopyridazine (200 mg, 0.728 mmol, 1.00 equiv) and tert-butyl 3-ethynylazetidine-1-carboxylate (145.06 mg, 0.801 mmol, 1.1 equiv) in toluene (5.00 mL) was added Pd(PPh3)2Cl2 (76.61 mg, 0.109 mmol, 0.15 equiv), CuI (27.71 mg, 0.146 mmol, 0.2 equiv) and TEA (220.88 mg, 2.184 mmol, 3 equiv) at room temperature under nitrogen atmosphere. The resulting mixture was stirred at room temperature for 2 hours and then filtered. The filtrate was concentrated under reduced pressure and the residue was purified by reverse phase flash chromatography under the following conditions: column, C18; mobile phase, MeCN / water (0.05% FA), 40% to 60% gradient. This provided the title compound as a yellow solid (170 mg, 64.07%). LCMS (ESI) m / z: [M+H]+ = 328.

[1017] Step 2: Preparation of tert-butyl 3-{3-chlorothieno[2,3-c]pyridazin-6-yl}azetidine-1-carboxylate

[1018]

[1019] To a stirred mixture of tert-butyl 3-[2-(3,6-dichloropyridazin-4-yl)ethynyl]azetidine-1-carboxylate (160 mg, 0.488 mmol, 1.00 equiv) in NMP (5 mL) was added sodium hydrosulfide (32.80 mg, 0.586 mmol, 1.2 equiv) at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 100 °C for 2 hours, cooled and filtered. The filtrate was purified by reverse phase flash chromatography under the following conditions: column, C18; mobile phase, MeCN / water (0.05% FA), 30% to 50% gradient. This provided the title compound (122 mg, 71.43%) as a white solid. LCMS (ESI) m / z [M+H] + = 326.

[1020] Step 3: Preparation of tert-butyl 3-[3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl]azetidine-1-carboxylate

[1021]

[1022] To a stirred mixture of tert-butyl 3-{3-chlorothieno[2,3-c]pyridazin-6-yl}azetidine-1-carboxylate (122 mg, 0.374 mmol, 1.00 equiv) and 2-hydroxyphenylboronic acid (154.94 mg, 1.122 mmol, 3 equiv) in dioxane (4 mL) and H2O (1 mL) was added Cs2CO3 (244.01 mg, 0.748 mmol, 2 equiv) and XPhos Pd G3 (63.39 mg, 0.075 mmol, 0.2 equiv) at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 80 °C for 2 hours and then filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18; mobile phase, MeCN / water (0.05% FA), 40% to 60% gradient. This provided the title compound as a white solid (85 mg, 53.28%). LCMS (ESI) m / z: [M+H]+ = 384.

[1023] Step 4: Preparation of 2-[6-(azetidin-3-yl)thieno[2,3-c]pyridazin-3-yl]phenol

[1024]

[1025] To a stirred mixture of tert-butyl 3-[3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl]azetidine-1-carboxylate (85 mg, 0.222 mmol, 1.00 equiv) in DCM (4 mL) was added TFA (1 mL) dropwise at room temperature under nitrogen atmosphere. The resulting mixture was stirred for 2 hours and then concentrated under reduced pressure. The crude product was used directly in the next step without further purification. LCMS (ESI) m / z: [M+H]+=284.

[1026] Preparation of 2-(6-(piperidin-4-yl)thieno[2,3-c]pyridazin-3-yl)phenol (I-7)

[1027]

[1028] Step 1: Preparation of tert-butyl 4-((3,6-dichloropyridazin-4-yl)ethynyl)piperidine-1-carboxylate (Intermediate 2)

[1029]

[1030] To a stirred mixture of tert-butyl 4-ethynylpiperidine-1-carboxylate (1.00 g, 4.778 mmol, 1.00 eq) and 4-bromo-3,6-dichloropyridazine (1.20 g, 5.256 mmol, 1.1 eq) in toluene (10.00 mL) was added Pd(PPh3)2Cl2 (0.50 g, 0.717 mmol, 0.15 eq), CuI (0.18 g, 0.956 mmol, 0.2 eq) and TEA (1.45 g, 14.334 mmol, 3 eq) at room temperature under nitrogen atmosphere. The resulting mixture was stirred at room temperature overnight and then concentrated under reduced pressure. The residue was dissolved in DMF (15.00 mL) and purified by reverse phase flash chromatography under the following conditions: column, C18; mobile phase, MeCN / water (0.05% FA), 0% to 100% gradient in 25 minutes to provide intermediate 2 (596 mg, 33.61%) as a brown solid. LCMS (ESI) m / z: [M+H]+=356.25

[1031] Step 2: Preparation of tert-butyl 4-(3-chlorothieno[2,3-c]pyridazin-6-yl)piperidine-1-carboxylate (Intermediate 3)

[1032]

[1033] To a stirred solution of intermediate 2 (596.00 mg, 1.673 mmol, 1.00 equiv) in NMP (10.00 mL) was added NaSH (93.79 mg, 1.673 mmol, 1.0 equiv) at room temperature. The resulting mixture was stirred at 100 °C for 1 hour and then allowed to cool to room temperature. The resulting mixture was extracted with EtOAc (3x200 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The residue was dissolved in DMF (10.00 mL) and purified by reverse phase flash chromatography under the following conditions: column, C18; mobile phase, MeCN / water (0.05% FA), 0% to 100% gradient in 30 minutes; to provide intermediate 3 (356 mg, 49.91%) as a brown solid. LCMS (ESI) m / z: [M+H]+ = 353.87.

[1034] Step 3: Preparation of tert-butyl 4-(3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl)piperidine-1-carboxylate (Intermediate 4)

[1035]

[1036] To a solution of intermediate 3 (350.00 mg, 0.989 mmol, 1.00 eq) and 2-hydroxyphenylboronic acid (204.63 mg, 1.484 mmol, 1.5 eq) in dioxane (5.00 mL) and H2O (1.00 mL) were added Cs2CO3 (644.51 mg, 1.978 mmol, 2.0 eq) and XPhos Pd G3 (83.72 mg, 0.099 mmol, 0.1 eq). After stirring at 90°C under nitrogen atmosphere overnight, the mixture was concentrated under reduced pressure. The residue was dissolved in DMF (10.00 mL) and purified by reverse phase flash chromatography under the following conditions: column, C18; mobile phase, MeCN / water (0.05% FA), 0% to 100% gradient in 20 minutes; to provide intermediate 4 (188 mg, 44.85%) as a brown solid. LCMS (ESI) m / z: [M+H]+=411.52.

[1037] Step 4: Preparation of 2-(6-(piperidin-4-yl)thieno[2,3-c]pyridazin-3-yl)phenol (I-7)

[1038]

[1039] To a stirred solution of intermediate 4 (188.00 mg, 0.457 mmol, 1.00 equiv) in DCM (9.00 mL) was added TFA (3.00 mL) at room temperature. The resulting mixture was stirred for 4 hours and then concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18; mobile phase, MeCN / water (0.05 FA), 0% to 100% gradient in 20 minutes; to provide the title compound (130 mg, 91.38%) as a light brown solid. LCMS (ESI) m / z: [M+H]+=311.40.

[1040] Preparation of 2-(5-methyl-6-(piperidin-4-yl)thieno[2,3-c]pyridazin-3-yl)phenol (I-8)

[1041]

[1042] Step 1: Preparation of tert-butyl 4-((3,6-dichloropyridazin-4-yl)ethynyl)piperidine-1-carboxylate (Intermediate 2)

[1043]

[1044] To a mixture of tert-butyl 4-ethynylpiperidine-1-carboxylate (8.00 g, 38.225 mmol, 1.00 equiv) and 4-bromo-3,6-dichloropyridazine (10.45 g, 45.870 mmol, 1.20 equiv) in toluene (80 mL) was added Pd(PPh3)2Cl2 (4.02 g, 5.734 mmol, 0.15 equiv), CuI (14.56 g, 76.450 mmol, 2.00 equiv) and TEA (11.60 g, 114.675 mmol, 3.00 equiv) under a nitrogen atmosphere. The resulting mixture was stirred at room temperature overnight and concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with 0% to 50% EtOAc / petroleum ether to provide intermediate 2 (5.00 g, 36.7%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=356.

[1045] Step 2: Preparation of tert-butyl 4-(3-chlorothieno[2,3-c]pyridazin-6-yl)piperidine-1-carboxylate (Intermediate 3)

[1046]

[1047] To a mixture of intermediate 2 (5.00 g, 14.035 mmol, 1.00 equiv) in NMP (50 mL) was added NaSH (0.79 g, 14.035 mmol, 1.0 equiv). The resulting mixture was stirred at 100 °C for 1 hour, cooled and filtered. The filtrate was purified by reverse phase flash chromatography under the following conditions: column, C18; mobile phase, MeOH / water, 10% to 50% gradient in 10 minutes; to provide intermediate 3 (1.80 g, 36.2%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=354.

[1048] Step 3: Preparation of tert-butyl 4-(5-bromo-3-chlorothieno[2,3-c]pyridazin-6-yl)piperidine-1-carboxylate (Intermediate 4)

[1049]

[1050] To a mixture of intermediate 3 (1.80 g, 5.087 mmol, 1.00 equiv) in CHCl3 (20 mL) was added Br2 (8.13 g, 50.870 mmol, 10.00 equiv). The resulting mixture was stirred at room temperature overnight and then basified with aqueous NaHCO3. Boc2O (2.21 g, 10.174 mmol, 2.00 equiv) was then added and the mixture was stirred for 2 hours. The mixture was extracted with EtOAc (3 x 100 mL), and the combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18; mobile phase, MeOH / water, 10% to 50% gradient in 10 minutes; to provide intermediate 4 (710.0 mg, 32.4%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=432.

[1051] Step 4: Preparation of tert-butyl 4-(3-chloro-5-methylthieno[2,3-c]pyridazin-6-yl)piperidine-1-carboxylate (Intermediate 5)

[1052]

[1053] To a mixture of intermediate 4 (710.0 mg, 1.641 mmol, 1.00 equiv), K3PO4 (696.50 mg, 3.282 mmol, 2.00 equiv) and Pd(AMPhos)Cl2 (174.25 mg, 0.246 mmol, 0.15 equiv) in dioxane (10 mL) and H2O (2 mL) was added trimethyl-1,3,5,2,4,6-trioxatriborinane (411.90 mg, 3.282 mmol, 2.00 equiv) and the resulting mixture was stirred at 60° C. for 1 hour under nitrogen atmosphere. The reaction mixture was filtered through a short pad of celite and eluted with EtOAc. The filtrate was concentrated under vacuum and the residue was purified by reverse phase flash chromatography under the following conditions: column, C18; mobile phase, MeOH / water, 10% to 50% gradient in 10 minutes; to provide intermediate 5 (450.0 mg, 74.5%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=368.

[1054] Step 5: Preparation of tert-butyl 4-(3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl)piperidine-1-carboxylate (Intermediate 6)

[1055]

[1056] To a mixture of intermediate 5 (450.0 mg, 1.223 mmol, 1.00 equiv) and 2-hydroxyphenylboronic acid (337.43 mg, 2.446 mmol, 2.00 equiv) in dioxane (10 mL) and H2O (2 mL) were added XPhos Pd G3 (155.31 mg, 0.183 mmol, 0.15 equiv) and Cs2CO3 (1.2 g, 3.669 mmol, 3.00 equiv) and the resulting mixture was stirred under nitrogen atmosphere at 100° C. for 1 hour. The reaction mixture was filtered through a short pad of celite and eluted with EtOAc. The filtrate was concentrated under vacuum and the residue was purified by reverse phase flash chromatography under the following conditions: column, C18; mobile phase, MeOH / water, 10% to 50% gradient in 10 minutes; to provide intermediate 6 (380.0 mg, 73.0%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=426.

[1057] Step 6: Preparation of 2-(5-methyl-6-(piperidin-4-yl)thieno[2,3-c]pyridazin-3-yl)phenol (I-8)

[1058]

[1059] To a mixture of intermediate 6 (380.0 mg, 0.893 mmol, 1.00 equiv) in DCM (6 mL) was added TFA (3 mL, 40.389 mmol, 45.23 equiv), and the resulting mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by reverse phase flash chromatography under the following conditions: column, C18; mobile phase, MeOH / water, 10% to 50% gradient in 10 minutes; detector to provide I-8 (246.4 mg, 84.7%) as a yellow solid. 1HNMR(400MHz,DMSO-d6)δ8.69(s,1H),8.22–8.11(m,1H),7.43–7.34(m,1H),7.07–6.99(m,2H),3.67–3.53(m,1H),3. 43–3.35(m,2H),3.10–3.00(m,2H),2.46(s,3H),2.13–2.04(m,2H),1.91–1.78(m,2H).LCMS(ESI)m / z:[M+H]+=326.10.

[1060] Preparation of methyl 2-[3-(6-fluoro-5-methylpyridin-3-yl)-1,2-oxazol-5-yl]-3-methylbutanoate (I-9)

[1061]

[1062] Step 1: Preparation of (Z)-N-[(6-fluoro-5-methylpyridin-3-yl)methylidene]hydroxylamine (Intermediate 2)

[1063]

[1064] To a solution of 6-fluoro-5-methylpyridine-3-carbaldehyde (10 mg, 0.072 mmol, 1 eq.), hydroxylamine hydrochloride (9.99 mg, 0.144 mmol, 2 eq.) and Na2CO3 (22.85 mg, 0.216 mmol, 3 eq.) in MeOH (0.5 mL) and H2O (0.5 mL), the resulting solution was stirred at 25°C overnight. The mixture was diluted with EtOAc (200 mL) and washed with water (200 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give intermediate 2 (2.2 mg, 99.29%) as a yellow solid. LCMS (ESI) m / z: [M+H]+ = 155.

[1065] Step 2: Preparation of (E)-6-fluoro-N-hydroxy-2-methylpyridine-3-carboximidoyl chloride (Intermediate 3)

[1066]

[1067] To a solution of intermediate 2 (2.2 g, 14.272 mmol, 1 eq.), NCS (2.86 g, 21.408 mmol, 1.5 eq.) in EtOAc (20 mL), the resulting solution was stirred at 25 °C overnight. The mixture was diluted with EtOAc (100 mL) and washed with water (100 mL x 2). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give intermediate 3 (3.6 g, crude) as a white solid. LCMS (ESI) m / z: [M+H]+=189.

[1068] Step 3: Preparation of methyl 2-[3-(6-fluoro-5-methylpyridin-3-yl)-1,2-oxazol-5-yl]acetate (Intermediate 4)

[1069]

[1070] To a solution of intermediate 3 (3.6 g, 19.089 mmol, 1 eq.) in EtOAc (14 mL) at 0°C was added but-3-ynoic acid methyl ester (3.75 g, 38.178 mmol, 2 eq.) and NaHCO3 (4.81 g, 57.267 mmol, 3 eq.). The resulting solution was stirred at 25°C overnight. The mixture was diluted with EtOAc (100 mL) and washed with water (100 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN / H2O, 0% to 100% gradient in 30 minutes to give intermediate 4 (2.5 g, 52.34%) as a white solid. LCMS (ESI) m / z: [M+H]+=251.

[1071] Step 4: Preparation of methyl 2-[3-(6-fluoro-5-methylpyridin-3-yl)-1,2-oxazol-5-yl]-3-methylbutanoate (I-9)

[1072]

[1073] To a solution of intermediate 4 (500 mg, 1.998 mmol, 1 eq.), 2-iodopropane (679.35 mg, 3.996 mmol, 2 eq.), Cs2CO3 (1.3 g, 3.996 mmol, 2 eq.) in THF (5 mL), the resulting solution was stirred at 60 °C overnight. The mixture was diluted with EtOAc (100 mL) and washed with water (100 mL x 2). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by flash C18 chromatography with an elution gradient of 0 to 60% ACN / H2O to give the title compound (288 mg, 49.31%) as a white solid. LCMS (ESI) m / z: [M+H]+=293.

[1074] Preparation of 2-(5-cyclopropyl-6-(2,6-diazaspiro[3.3]hept-2-yl)thieno[2,3-c]pyridazin-3-yl)phenol (I-11)

[1075]

[1076] Step 1: Preparation of tert-butyl 2-cyano-2-cyclopropylacetate (Intermediate 2)

[1077]

[1078] To a solution of 2-cyclopropylacetonitrile (10.00 g, 123.277 mmol, 1 eq.) and Boc2O (53.81 g, 246.554 mmol, 2 eq.) in THF (100 mL) was added LDA (26.41 g, 246.554 mmol, 2 eq.) at -78 °C. After stirring for 2 hours at -78 °C, the reaction was quenched with water (100 mL) at 0 °C. The resulting mixture was extracted with EtOAc (100 mL x 3). The combined organic layer was washed with brine (100 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with 0% to 20% EtOAc / PE to provide intermediate 2 (20.00 g, 80.57%) as a yellow oil. It was confirmed by H-NMR that there was no Ms signal.

[1079] Step 2: tert-Butyl 2-cyano-2-cyclopropyl-2-(3,6-dichloropyridazin-4-yl)acetate (Intermediate 3)

[1080]

[1081] To a solution of intermediate 2 (15.00 g, 82.765 mmol, 1 eq.) and 3,4,6-trichloropyridazine (15.18 g, 82.765 mmol, 1 eq.) in DMSO (100 mL) was added DIEA (32.09 g, 248.295 mmol, 3 eq.). After stirring overnight at room temperature under a nitrogen atmosphere, the resulting mixture was diluted with water (500 mL). The resulting mixture was extracted with EtOAc (500 mL x 3). The combined organic layers were washed with brine (500 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with 0% to 50% EtOAc / PE to provide intermediate 3 (18.00 g, 59.64%) as a pink solid. LCMS (ESI) m / z: [M+H]+=328.

[1082] Step 3: Preparation of 2-cyclopropyl-2-(3,6-dichloropyridazin-4-yl)acetonitrile (Intermediate 4)

[1083]

[1084] To a solution of intermediate 3 (10.00 g, 30.470 mmol, 1 eq.) in DMSO (50 mL) and H2O (5 mL) was added NaCl (3.56 g, 60.940 mmol, 2 eq.). After stirring at 100 °C for 3 hours, the resulting mixture was diluted with water (500 mL). The resulting mixture was extracted with EtOAc (500 mL x 3). The combined organic layers were washed with brine (500 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with 0% to 50% EtOAc / PE to provide intermediate 4 (5.00 g, 64.7%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=228.

[1085] Step 4: Preparation of 3-chloro-5-cyclopropylthieno[2,3-c]pyridazin-6-amine (Intermediate 5)

[1086]

[1087] To a solution of intermediate 4 (3.00 g, 13.153 mmol, 1 eq.) in NMP (50 mL) was added NaHS.2H2O (1.21 g, 13.153 mmol, 1 eq.). After stirring at 100 °C for 20 minutes, the resulting mixture was diluted with water (500 mL). The resulting mixture was extracted with EtOAc (200 mL x 3). The combined organic layers were washed with brine (200 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with 0% to 50% EtOAc / PE to provide intermediate 5 (2.00 g, 60.6%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=226.

[1088] Step 5: Preparation of 3,6-dichloro-5-cyclopropylthieno[2,3-c]pyridazine (Intermediate 6)

[1089]

[1090] To a solution of intermediate 5 (2.00 g, 8.862 mmol, 1 eq.) and CuCl2 (2.38 g, 17.724 mmol, 2 eq.) in ACN (50 mL) was added t-BuNO2 (1.83 g, 17.724 mmol, 2 eq.). After stirring at 50 °C for 3 hours, the resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with 0% to 50% EtOAc / PE to provide intermediate 6 (1.00 g, 41.4%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=245.

[1091] Step 6: Preparation of tert-butyl 6-(3-chloro-5-cyclopropylthieno[2,3-c]pyridazin-6-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (Intermediate 7)

[1092]

[1093] To a solution of intermediate 6 (1.00 g, 4.080 mmol, 1 eq.) and tert-butyl 2,6-diazaspiro [3.3] heptane-2-carboxylate (1.62 g, 8.160 mmol, 2 eq.) in DMSO (20 mL) was added DIEA (1.58 g, 12.240 mmol, 3 eq.). After stirring at 100 ° C for 3 hours under a nitrogen atmosphere, the resulting mixture was diluted with water (100 mL). The resulting mixture was extracted with EtOAc (100 mL x 3). The combined organic layer was washed with brine (200 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with 0% to 50% EtOAc / PE to provide intermediate 7 (900.0 mg, 48.7%) as a yellow solid. LCMS (ESI) m / z: [M+H]+ = 407.

[1094] Step 7: Preparation of tert-butyl 6-(5-cyclopropyl-3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (Intermediate 7)

[1095]

[1096] To a solution of intermediate 7 (900.0 mg, 2.212 mmol, 1 eq.) and 2-hydroxyphenylboronic acid (610.11 mg, 4.424 mmol, 2 eq.) in dioxane (20 mL) and H2O (4 mL) was added Cs2CO3 (2161.8 mg, 6.636 mmol, 3 eq.) and XPhos Pd G3 (187.2 mg, 0.221 mmol, 0.1 eq.). After stirring at 80 °C for 1 hour under a nitrogen atmosphere, the resulting mixture was diluted with water (50 mL). The resulting mixture was extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (100 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN / water, 10% to 50% gradient in 10 minutes; detector, UV 254 nm to provide intermediate 8 (800.0 mg, 70.0%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=465.

[1097] Step 8: Preparation of 2-(5-cyclopropyl-6-(2,6-diazaspiro[3.3]hept-2-yl)thieno[2,3-c]pyridazin-3-yl)phenol (I-11)

[1098]

[1099] To a solution of intermediate 8 (500.0 mg, 1.076 mmol, 1 eq.) in DCM (9 mL) was added TFA (3 mL, 40.389 mmol, 37.53 eq.). After stirring at room temperature for 1 hour, the resulting mixture was concentrated under reduced pressure to provide I-11 (700.0 mg, crude) as a semisolid. The crude product was used directly in the next step without further purification. LCMS (ESI) m / z: [M+H]+=365.

[1100] Preparation of 2-(6-{2,6-diazaspiro[3.3]hept-2-yl}-5-methylthieno[2,3-c]pyridazin-3-yl)phenol (I-12)

[1101]

[1102] Step 1: Preparation of tert-butyl 6-{3-chloro-5-methylthieno[2,3-c]pyridazin-6-yl}-2,6-diazaspiro[3.3]heptane-2-carboxylate (Intermediate 2)

[1103]

[1104] To a solution of 3,6-dichloro-5-methylthieno[2,3-c]pyridazine (639.12 mg, 2.916 mmol, 2.41 equiv) in DMSO (8 mL) was added tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate (240 mg, 1.210 mmol, 1.00 equiv) and DIEA (469.36 mg, 3.630 mmol, 3 equiv). The resulting mixture was stirred at 60 °C for 6 hours under nitrogen atmosphere. The desired product can be detected by LCMS. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.1% FA), 10% to 80% gradient in 30 minutes; detector, UV 254 nm. This produced intermediate 2 (280 mg, 42.51%) as a yellow oil. LCMS (ESI) m / z: [M+H]+=381.

[1105] Step 2: Preparation of tert-butyl 6-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]-2,6-diazaspiro[3.3]heptane-2-carboxylate (Intermediate 3)

[1106]

[1107] To a solution of tert-butyl 6-{3-chloro-5-methylthieno[2,3-c]pyridazine-6-yl}-2,6-diazaspiro[3.3]heptane-2-carboxylate (300 mg, 0.788 mmol, 1 eq) and 2-hydroxyphenylboronic acid (217.28 mg, 1.576 mmol, 2 eq) in dioxane (6 mL) and H2O (1 mL) was added Cs2CO3 (769.87 mg, 2.364 mmol, 3 eq) and XPhos PdG3 (66.67 mg, 0.079 mmol, 0.1 eq). The resulting mixture was stirred at 80 °C for 3 hours under nitrogen atmosphere. The resulting mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography eluting with PE / EA (1:1) to give intermediate 3 (160 mg, 46.32%) as a yellow solid. LCMS (ESI) m / z [M+H] + = 439.

[1108] Step 3: Preparation of 2-(6-{2,6-diazaspiro[3.3]hept-2-yl}-5-methylthieno[2,3-c]pyridazin-3-yl)phenol (I-12)

[1109]

[1110] A solution of intermediate 3 (150 mg, 0.342 mmol, 1 eq.) in TFA (1 mL) and DCM (3 mL) was stirred at room temperature for 1 hour. The desired product can be detected by LCMS. The resulting mixture was concentrated under vacuum. This yielded I-12 (100 mg, 86.39%) as a yellow oil. LCMS (ESI) m / z: [M+H]+=339.

[1111] Preparation of 2-(6-{2,6-diazaspiro[3.3]hept-2-yl}thieno[3,2-c]pyridazin-3-yl)phenol) (I-13)

[1112]

[1113] Step 1: Preparation of 4-bromo-6-chloro-3-iodopyridazine (Intermediate 2)

[1114]

[1115] To a stirred solution of 4-bromo-6-chloropyridazin-3-amine (43.00 g, 206.294 mmol, 1 eq.) and CH2I2 (66.30 g, 247.553 mmol, 1.2 eq.) in THF (300 mL) was added CuI (47.15 g, 247.553 mmol, 1.2 eq.) and t-BuNO2 (25.53 g, 247.553 mmol, 1.2 eq.). The resulting mixture was stirred at 60 °C overnight. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with PE / EtOAc (10:1) to provide intermediate 2 (25.00 g, 58.1%) as a light yellow solid. LCMS (ESI) m / z: [M+H]+=319.

[1116] Step 2: Preparation of 4-(tert-butylsulfanyl)-6-chloro-3-iodopyridazine (Intermediate 3)

[1117]

[1118] To a stirred solution of intermediate 2 (25.00 g, 78.291 mmol, 1 eq.) and 2-methyl-2-propanethiol (7.77 g, 86.120 mmol, 1.1 eq.) in DMF (300 mL) was added Cs2CO3 (76.3 g, 234.873 mmol, 3 eq.). The resulting mixture was stirred at 100 °C for 1 hour. The resulting mixture was concentrated under reduced pressure. The resulting mixture was diluted with water (300 mL). The resulting mixture was extracted with EtOAc (3 x 500 mL). The combined organic layer was washed with brine (1 x 200 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with PE / EtOAc (10: 1) to provide intermediate 3 (16.00 g, 62.2%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=329.

[1119] Step 3: Preparation of tert-butyl 2-[4-(tert-butylsulfanyl)-6-chloropyridazin-3-yl]-2-cyanoacetate (Intermediate 4)

[1120]

[1121] A solution of intermediate 3 (16.00 g, 48.691 mmol, 1 eq.) and tert-butyl 2-cyanoacetate (13.75 g, 97.382 mmol, 2 eq.) and Cs2CO3 (47.4 g, 146.073 mmol, 3 eq.) in 1,4-dioxane was stirred at room temperature for 30 minutes. To the above mixture was added picolinic acid (3.00 g, 24.346 mmol, 0.5 eq.). The resulting mixture was stirred at 80 °C for another 2 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with DCM / MeOH (10:1) to provide intermediate 4 (5.30 g, 33.1%) as a brown solid. LCMS (ESI) m / z: [M+H]+=342.

[1122] Step 4: Preparation of 3-chlorothieno[3,2-c]pyridazin-6-amine (Intermediate 5)

[1123]

[1124] A solution of intermediate 4 (5.3 g, 15.504 mmol, 1 eq.) and 6N HCl (50 mL) in AcOH (50 mL) was stirred at 80 ° C overnight. The resulting mixture was concentrated under reduced pressure. The residue was neutralized to pH 7 with saturated sodium bicarbonate solution. The resulting mixture was extracted with EtOAc (3x 300 mL). The combined organic layers were washed with brine (2x200 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. This produced intermediate 5 (3.60 g, crude) as a yellow solid. The crude product was used directly in the next step without further purification. LCMS (ESI) m / z: [M+H] + = 186.

[1125] Step 5: Preparation of 3,6-dichlorothieno[3,2-c]pyridazine (Intermediate 6)

[1126]

[1127] To a stirred solution of intermediate 5 (3.60 g, 19.393 mmol, 1 eq.) and CuCl (3.84 g, 38.786 mmol, 2 eq.) in MeCN (50 mL) was added tert-butyl nitrite (4.00 g, 38.789 mmol, 2 eq.) in portions at 0°C. The resulting mixture was stirred at 50°C for 2 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (1:1) to provide intermediate 6 (543.0 mg, 15.0%) as a light brown solid. LCMS (ESI) m / z: [M+H]+=205.

[1128] Step 6: Preparation of tert-butyl 6-(3-chlorothieno[3,2-c]pyridazin-6-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (Intermediate 7)

[1129]

[1130] To a stirred solution of intermediate 6 (543.0 mg, 2.648 mmol, 1 eq.) and tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate (525.01 mg, 2.648 mmol, 1 eq.) in DMSO (8 mL) was added DIEA (1.03 g, 7.944 mmol, 3 eq.). The resulting mixture was stirred at 100 °C for 1 hour. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.1% FA), 10% to 50% gradient in 10 minutes; detector, UV 254 nm. This produced intermediate 7 (253.0 mg, 46.5%) as a brownish yellow solid. LCMS (ESI) m / z: [M+H]+ = 367.

[1131] Step 7: Preparation of tert-butyl 6-[3-(2-hydroxyphenyl)thieno[3,2-c]pyridazin-6-yl]-2,6-diazaspiro[3.3]heptane-2-carboxylate (Intermediate 8)

[1132]

[1133] To a stirred solution of intermediate 7 (253.0 mg, 0.690 mmol, 1 eq.) and 2-(methoxymethoxy)phenylboronic acid (125.5 mg, 0.690 mmol, 1 eq.) in 1,4-dioxane (4 mL) and H2O (1 mL) was added K3PO4 (292.7 mg, 1.380 mmol, 2 eq.) and (DiMeIHeptCl)Pd(cinnamoyl)Cl (80.5 mg, 0.069 mmol, 0.1 eq.). The resulting mixture was stirred for 2 hours and concentrated under reduced pressure. The resulting mixture was diluted with water (50 mL). The resulting mixture was extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (1 x 50 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.1% FA), 10% to 50% gradient in 10 minutes; detector, UV 254 nm. This produced intermediate 8 (98.0 mg, 41.8%) as a brown solid. LCMS (ESI) m / z: [M+H]+ = 469.

[1134] Step 8: Preparation of 2-(6-{2,6-diazaspiro[3.3]hept-2-yl}thieno[3,2-c]pyridazin-3-yl)phenol) (I-13)

[1135]

[1136] A solution of intermediate 8 (98.0 mg, 0.213 mmol, 1 eq.) and TFA (0.30 mL, 4.039 mmol, 19.31 eq.) in DCM (0.9 mL) was stirred at room temperature for 30 minutes. The resulting mixture was concentrated under reduced pressure. This produced I-13 (106.0 mg, crude) as a light red oil. The crude product was used directly in the next step without further purification. LCMS (ESI) m / z: [M+H]+=325

[1137] Preparation of (2S,4R)-4-hydroxy-1-[(2R)-2-(3-{4-[2-(4-{3-[(3Z)-2-hydroxypenta-1,3-dien-3-yl]-5-methylthieno[2,3-c]pyridazin-6-yl}piperidin-1-yl)pyrimidin-5-yl]piperazin-1-yl}-1,2-oxazol-5-yl)-3-methylbutanoyl]-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (Compound 14)

[1138]

[1139] Step 1: Preparation of tert-butyl 4-(2-methoxypyrimidin-5-yl)piperazine-1-carboxylate (Intermediate 2)

[1140]

[1141] To a solution of 5-bromo-2-methoxypyrimidine (9 g, 47.616 mmol, 1 eq.), tert-butyl piperazine-1-carboxylate (13.30 g, 71.424 mmol, 1.5 eq.), Pd2(dba)3 (2.18 g, 2.381 mmol, 0.05 eq.), BINAP (2.96 g, 4.762 mmol, 0.1 eq.) in toluene (75 mL), the resulting solution was stirred at 80 °C for 6 hours. The desired product can be detected by LCMS. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN / H2O, 0% to 25% gradient in 30 minutes to obtain intermediate 2 (4.2 g, 29.97%) as a white solid. LCMS (ESI) m / z: [M+H]+=295.

[1142] Step 2: Preparation of 2-methoxy-5-(piperazin-1-yl)pyrimidine (Intermediate 3)

[1143]

[1144] To a solution of intermediate 2 (4.2 g, 14.268 mmol, 1 eq.) in TFA (10 mL) and DCM (30 mL) was added and the resulting solution was stirred at 25 °C for 5 hours. The DCM in the reaction solution was spun off in vacuo to give crude intermediate 3 (10.0 g, crude) as a yellow oil. LCMS (ESI) m / z: [M+H]+=195.

[1145] Step 3: Preparation of methyl 2-{3-[4-(2-methoxypyrimidin-5-yl)piperazin-1-yl]-1,2-oxazol-5-yl}-3-methylbutanoate (Intermediate 4)

[1146]

[1147] To a solution of intermediate 3 (10 g, 51.483 mmol, 1 eq.), methyl 3-methyl-2-{3-[(1,1,2,2,3,3,4,4,4-nonafluorobutanesulfonyl)oxy]-1,2-oxazol-5-yl}butanoate (24.78 g, 51.483 mmol, 1 eq.) and DIEA (19.96 g, 154.449 mmol, 3 eq.) in DMSO (40 mL), the resulting solution was stirred at 100 °C overnight. The desired product can be detected by LCMS. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 0 to 30% gradient in 30 minutes; detector, UV 220 / 200 nm to give intermediate 4 (3.7 g, 19.14%). LCMS (ESI) m / z: [M+H]+=376.

[1148] Step 4: Preparation of methyl 2-{3-[4-(2-chloropyrimidin-5-yl)piperazin-1-yl]-1,2-oxazol-5-yl}-3-methylbutanoate (Intermediate 5)

[1149]

[1150] To a solution of intermediate 4 (900 mg, 2.397 mmol, 1 eq.) in POCl3 (6 mL) was added the resulting solution, which was stirred at 110°C for 18 hours. The desired product can be detected by LCMS. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.1% FA), 0 to 58% gradient in 30 minutes; detector, UV 220 / 200 nm. This produced intermediate 5 (360 mg, 39.53%) as a yellow solid.

[1151] Step 5: Preparation of methyl 2-{3-[4-(2-{4-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]piperidin-1-yl}pyrimidin-5-yl)piperazin-1-yl]-1,2-oxazol-5-yl}-3-methylbutanoate (Intermediate 6)

[1152]

[1153] To a solution of intermediate 5 (200 mg, 0.527 mmol, 1 eq.) and 2-[5-methyl-6-(piperidin-4-yl)thieno[2,3-c]pyridazin-3-yl]phenol (342.69 mg, 1.054 mmol, 2 eq.) in dioxane (4 mL) was added Cs2CO3 (343.10 mg, 1.054 mmol, 2 eq.) and Pd-PEPPSI-IPentCl 2-methylpyridine (o-methylpyridine) (88.58 mg, 0.105 mmol, 0.2 eq.) and the resulting solution was stirred at 90°C overnight. The desired product can be detected by LCMS. The residue was purified by reverse phase flash chromatography under the following conditions: column, silica gel; mobile phase, PE / EA, 0% to 56% to afford intermediate 6 (100 mg, 28.4%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=668.

[1154] Step 6: Preparation of 2-{3-[4-(2-{4-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]piperidin-1-yl}pyrimidin-5-yl)piperazin-1-yl]-1,2-oxazol-5-yl}-3-methylbutanoic acid (Intermediate 7)

[1155]

[1156] To a solution of methyl intermediate 6 (95 mg, 0.142 mmol, 1 eq.) in MeOH (4 mL) and H2O (1 mL) was added LiOH (27.22 mg, 1.136 mmol, 8 eq.) and the resulting solution was stirred at room temperature for 16 hours. The mixture was acidified to pH 5 with 1M HCl (aq.). The mixture was diluted with EtOAc (50 mL) and washed with water (50 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give intermediate 7 (98 mg, crude) as a yellow solid, which was used directly in the next step without further purification. LCMS (ESI) m / z: [M+H]+=655.

[1157] Step 7: Preparation of (2S,4R)-4-hydroxy-1-(2-{3-[4-(2-{4-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]piperidin-1-yl}pyrimidin-5-yl)piperazin-1-yl]-1,2-oxazol-5-yl}-3-methylbutanoyl)-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (Intermediate 8)

[1158]

[1159] To a solution of intermediate 7 (98 mg, 0.150 mmol, 1 eq), (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (99.21 mg, 0.300 mmol, 2 eq) and PyBOP (155.77 mg, 0.300 mmol, 2 eq) in DMF (2 mL) was stirred at 25 °C for 10 min, then DIEA (96.72 mg, 0.750 mmol, 5 eq) was added to the mixture. The resulting solution was stirred at 25 °C for 2 h. The reaction was purified by flash C18 chromatography with an elution gradient of 0 to 83% ACN / H2O to afford intermediate 8 (87 mg, 59.28%) as a yellow solid. LCMS (ESI) m / z: [M+H]+ = 968.

[1160] Step 8: Preparation of (2S,4R)-4-hydroxy-1-[(2R)-2-(3-{4-[2-(4-{3-[(3Z)-2-hydroxypenta-1,3-dien-3-yl]-5-methylthieno[2,3-c]pyridazin-6-yl}piperidin-1-yl)pyrimidin-5-yl]piperazin-1-yl}-1,2-oxazol-5-yl)-3-methylbutanoyl]-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide

[1161]

[1162] Intermediate 8 (94 mg) was purified by CHIRAL-HPLC using the following conditions (column: CHIRAL ART Cellulose-SB, 2*25 cm, 5 μm; mobile phase A: MtBE (10 mM NH3-MeOH), mobile phase B: MeOH--HPLC; flow rate: 20 mL / min; gradient: 20% B to 20% B in 20 minutes; wavelength: 270 / 212 nm; RT1 (min): 10.4; RT2 (min): 14.3; sample solvent: MeOH--HPLC; injection volume: 0.5 mL; number of runs: 6 to provide the title compound (second peak) (37.6 mg, 25.62%) as an off-white solid. The obtained solid was dried by lyophilization. dry. 1HNMR (300MHz, DMSO-d6) δ12.79(s,1H),8.99(s,1H),8.71(s,1H),8.42(d,J=7.6Hz,1H),8.28(s,2H),8. 17(d,J=7.4Hz,1H),7.52–7.41(m,2H),7.40–7.33(m,3H),7.08–6.97(m,2H),6.25(s,1H),5.12(d,J=3.7 Hz,1H),5.01–4.86(m,1H),4.75(d,J=13.1Hz,2H),4.38(t,J=7.8Hz,1H),4.30(s,1H),3.73(dd,J=10.4,4 .3Hz,1H),3.60(d,J=10.0Hz,2H),3.45(d,J=10.1Hz,1H),3.41–3.36(m,4H),3.16–2.97(m,6H),2.49(s,3 H),2.46(s,3H),2.24–2.19(m,1H),2.18–1.92(m,3H),1.87–1.78(m,1H),1.71–1.55(m,2H),1.47(d,J=7. 0Hz, 1H), 1.39 (d, J = 7.0Hz, 2H), 0.97 (d, J = 6.5Hz, 3H), 0.87–0.77 (m, 3H). LCMS (ESI) m / z: [M+H]+ = 968.40.

[1163] Preparation of (2S,4R)-4-hydroxy-1-[(2R)-2-(3-{2-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]ethynyl}-1,2-oxazol-5-yl)-3-methylbutanoyl]-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (Compound 37)

[1164]

[1165] Step 1: Preparation of (E)-N-[2-(Benzyloxy)ethylidene]hydroxylamine (Intermediate 2)

[1166]

[1167] To a stirred mixture of 2-(benzyloxy)acetaldehyde (10 g, 66.588 mmol, 1 eq.) and Na2CO3 (3.53 g, 33.294 mmol, 0.5 eq.) in EtOH (100 mL) was added NH2OH.HCl (5.09 g, 73.247 mmol, 1.1 eq.) at room temperature. The resulting mixture was stirred at room temperature for 2 hours. The desired product can be detected by LCMS. The resulting mixture was diluted with water (200 mL). The resulting mixture was extracted with EtOAc (3x200 mL). The combined organic layers were washed with brine (3x50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to provide intermediate 2 (13.196 g, crude) as a colorless liquid. LCMS (ESI) m / z: [M+H]+=166.

[1168] Step 2: Preparation of (Z)-2-(Benzyloxy)-N-hydroxyethanecarboximidoyl chloride (Intermediate 3)

[1169]

[1170] A mixture of intermediate 2 (12 g, 72.643 mmol, 1 eq.) and NCS (10.67 g, 79.907 mmol, 1.1 eq.) in DMF (100 mL) was stirred at room temperature for 2 hours. The desired product can be detected by LCMS. The resulting mixture was diluted with brine (200 mL). The resulting mixture was extracted with EtOAc (3x200 mL). The combined organic layer was washed with brine (3x50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to provide intermediate 3 (18.1 g, crude) as a light blue liquid. LCMS (ESI) m / z[M+H]+=200.

[1171] Step 3: Preparation of methyl 2-{3-[(benzyloxy)methyl]-1,2-oxazol-5-yl}acetate (Intermediate 4)

[1172]

[1173] A mixture of intermediate 3 (12 g, 60.111 mmol, 1 eq.) and NaHCO3 (7.57 g, 90.166 mmol, 1.5 eq.) in EA (100 mL) was stirred at room temperature for 30 min. Methyl but-3-ynoate (5.90 g, 60.111 mmol, 1 eq.) was added dropwise to the above mixture at 0°C. The resulting mixture was stirred for another overnight at room temperature. The desired product can be detected by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.1% FA), 0% to 100% gradient in 30 min; detector, UV 220 nm to provide intermediate 4 (8.3 g, crude) as a yellow liquid. LCMS (ESI) m / z: [M+H]+=262.

[1174] Step 4: Preparation of methyl 2-{3-[(benzyloxy)methyl]-1,2-oxazol-5-yl}-3-methylbutanoate (Intermediate 5)

[1175]

[1176] To a stirred mixture of intermediate 4 (8 g, 30.619 mmol, 1 eq.) and MgSO4 (7.37 g, 61.238 mmol, 2 eq.) in THF (80 mL) was added t-BuOK (15.31 mL, 15.309 mmol, 0.5 eq.) at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred at 0 °C under a nitrogen atmosphere for 30 minutes. 2-Iodopropane (6.25 g, 36.743 mmol, 1.2 eq.) was added to the above mixture at 0 °C. The resulting mixture was further stirred overnight at room temperature. The desired product can be detected by LCMS. The resulting mixture was filtered and the filter cake was washed with DMF (3x5 mL). The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.1% FA), 0% to 100% gradient in 30 minutes; detector, UV 220 nm to provide intermediate 5 (4.18 g, 45.00%) as a brown liquid. LCMS (ESI) m / z: [M+H]+=304.

[1177] Step 5: Preparation of methyl 2-[3-(hydroxymethyl)-1,2-oxazol-5-yl]-3-methylbutanoate (Intermediate 6)

[1178]

[1179] A mixture of intermediate 5 and BBr3 (9.91 g, 39.558 mmol, 3 equiv.) in DCM (40 mL) was stirred at 0 ° C under a nitrogen atmosphere for 2 hours. The desired product can be detected by LCMS. The reaction was quenched with water / ice at 0 ° C. The resulting mixture was extracted with CH2Cl2 (3x200 mL). The combined organic layer was washed with water (3x50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to provide intermediate 6 (1.5 g, pure) as a yellow oil. LCMS (ESI) m / z: [M+H]+=214.

[1180] Step 6: Preparation of methyl 2-(3-formyl-1,2-oxazol-5-yl)-3-methylbutanoate (Intermediate 7)

[1181]

[1182] A mixture of methyl intermediate 6 and Dess-Martin (1193.46 mg, 2.814 mmol, 1.2 equiv) in DCM (5 mL) was stirred at room temperature for 2 hours. The desired product can be detected by LCMS. The reaction was quenched at room temperature by adding NaHCO3 (aqueous solution) and Na2S2O3 (aqueous solution) (200 mL). The resulting mixture was extracted with EtOAc (3x 200 mL). The combined organic layer was washed with brine (3x30 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to provide intermediate 7 (530 mg, crude) as a light yellow liquid. LCMS (ESI) m / z: [M+H]+=212.

[1183] Step 7: Preparation of methyl 2-(3-ethynyl-1,2-oxazol-5-yl)-3-methylbutanoate (Intermediate 8)

[1184]

[1185] To a stirred mixture of intermediate 7 (500 mg, 2.367 mmol, 1 eq.) and K2CO3 (981.49 mg, 7.101 mmol, 3 eq.) in MeOH (5 mL) was added seyferth-gilberthomologation (682.16 mg, 3.550 mmol, 1.5 eq.) dropwise at 0 °C. The resulting mixture was stirred at room temperature overnight. The desired product can be detected by LCMS. The reaction was quenched with water at room temperature. The resulting mixture was extracted with CH2Cl2 (3 x 100 mL). The combined organic layers were washed with brine (3 x 10 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.1% FA), gradient from 0% to 100% in 30 minutes; detector, UV 254 nm to provide intermediate 8 (320 mg, 65.23%) as a light yellow oil. LCMS (ESI) m / z: [M+H]+=208.

[1186] Step 8: Preparation of methyl 2-[3-(2-{3-chloro-5-methylthieno[2,3-c]pyridazin-6-yl}ethynyl)-1,2-oxazol-5-yl]-3-methylbutanoate (Intermediate 9)

[1187]

[1188] A mixture of intermediate 8 (165.33 mg, 0.627 mmol, 1 eq), [1,3-bis[2,6-bis(propan-2-yl)phenyl]-2,3-dihydro-1H-imidazol-2-yl]dichloro(3-chloropyridin-1-ium-1-yl)palladium (42.62 mg, 0.063 mmol, 0.1 eq), CuI (11.95 mg, 0.063 mmol, 0.1 eq) and DIEA (405.40 mg, 3.135 mmol, 5 eq) in DMF (4 mL) was stirred at 80 °C under nitrogen atmosphere for 3 h. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.1% FA), 0% to 100% gradient in 30 minutes; detector, UV 254 nm to provide intermediate 9 (150 mg, 61.33%) as a brown solid. LCMS (ESI) m / z: [M+H]+=389.

[1189] Step 9: Preparation of methyl 2-(3-{2-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]ethynyl}-1,2-oxazol-5-yl)-3-methylbutanoate (Intermediate 10)

[1190]

[1191] A mixture of intermediate 9 (159.21 mg, 1.155 mmol, 3 eq.), XPhos PdG3 (65.14 mg, 0.077 mmol, 0.2 eq.) and Cs2CO3 (376.09 mg, 1.155 mmol, 3 eq.) in dioxane (3 mL) and H2O (0.6 mL) was stirred at 80 °C under a nitrogen atmosphere for 2 hours. The desired product can be detected by LCMS. The resulting mixture is concentrated under reduced pressure. The resulting mixture is diluted with water (200 mL). The resulting mixture is extracted with EtOAc (3x 200 mL). The combined organic layer is washed with brine (3x10 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate is concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.1% FA), 0% to 100% gradient in 30 minutes; detector, UV 254 nm to provide intermediate 10 (109 mg, 63.30%) as a light reddish brown solid. LCMS (ESI) m / z: [M+H]+=448.

[1192] Step 10: Preparation of 2-(3-{2-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]ethynyl}-1,2-oxazol-5-yl)-3-methylbutanoic acid (Intermediate 11)

[1193]

[1194] A mixture of intermediate 10 (100 mg, 0.223 mmol, 1 eq.) and LiOH.H2O (26.76 mg, 1.115 mmol, 5 eq.) in THF (3 mL) and H2O (3 mL) was stirred at room temperature for 2 hours. The desired product can be detected by LCMS. The mixture was acidified to pH 6 with concentrated HCl. The resulting mixture was concentrated under reduced pressure to provide intermediate 11 (400 mg, crude) as a brown solid. LCMS (ESI) m / z: [M+H]+=434.

[1195] Step 11: Preparation of (2S,4R)-4-hydroxy-1-[2-(3-{2-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]ethynyl}-1,2-oxazol-5-yl)-3-methylbutanoyl]-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (Intermediate 12)

[1196]

[1197] A mixture of intermediate 11 (100 mg, 0.231 mmol, 1 eq), (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (76.46 mg, 0.231 mmol, 1 eq), PyBOP (180.08 mg, 0.347 mmol, 1.5 eq) and DIEA (119.26 mg, 0.924 mmol, 4 eq) in DMF (2 mL) was stirred at room temperature for 2 hours. The desired product can be detected by LCMS. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (10 mmol / L NH4HCO3), 0% to 100% gradient in 30 minutes; detector, UV 254 nm to provide intermediate 12 (150 mg, 87.06%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=747.

[1198] Step 12: Preparation of (2S,4R)-4-hydroxy-1-[(2R)-2-(3-{2-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]ethynyl}-1,2-oxazol-5-yl)-3-methylbutanoyl]-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide

[1199]

[1200] The crude product intermediate 12 was purified by SFC-HPLC using the following conditions (column: CHIRALPAK IA, 3*25 cm, 5 μm; mobile phase A: CO2, mobile phase B: MeOH:DCM=1:1--HPLC; flow rate: 60 mL / min; gradient: isocratic 55% B; column temperature (°C): 35; back pressure (bar): 100; wavelength: 274 nm; RT1 (min): 4.27; RT2 (min): 7.20; sample solvent: MeOH:DCM=1:1--HPLC; injection volume: 1.6 mL) to provide the title compound (second peak) (38.1 mg, 38.73%) as a light yellow solid. 1HNMR(400MHz,DMSO-d6)δ12.09(s,1H),9.01–8.94(m,1H),8.88–8.81(m,1H),8.45(d,J=7.6Hz,1H),8.16–8.09(m,1H),7 .50–7.34(m,5H),7.09–7.00(m,2H),6.92(s,1H),5.14–5.09(m,1H),4.97–4.89(m,1H),4.40(t,J=8.0Hz,1H),4.30(s,1H ),3.97(d,J=9.6Hz,1H),3.77–3.47(m,2H),2.68–2.62(m,3H),2.47–2.45(m,3H),2.40–2.25(m,1H),2.10–2.01(m,1H),1 .85–1.74(m,1H),1.44(dd,J=37.1,7.0Hz,3H),1.01(d,J=6.4Hz,3H),0.88–0.79(m,3H).LCMS(ESI)m / z:[M+H]+=747.20.

[1201] Preparation of (2S,4R)-4-hydroxy-1-[(2R)-2-{3-[4-(2-{4-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]piperidin-1-yl}pyrimidin-5-yl)piperidin-1-yl]-1,2-oxazol-5-yl}-3-methylbutyryl]-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (Compound 60)

[1202]

[1203] Step 1: Preparation of tert-butyl 4-(2-methoxypyrimidin-5-yl)piperidine-1-carboxylate (Intermediate 2)

[1204]

[1205] To a solution of 5-bromo-2-methoxypyrimidine (10.0 g, 52.907 mmol, 1.00 equiv) and tert-butyl 4-iodopiperidine-1-carboxylate (19.8 g, 63.488 mmol, 1.20 equiv), dtbpy (1.4 g, 5.291 mmol, 0.10 equiv), Mn (5.8 g, 105.814 mmol, 2.00 equiv), KI (8.8 g, 52.907 mmol, 1 equiv), NiBr2.DME (1.9 g, 5.291 mmol, 0.10 equiv) in DMA (100.0 mL) was added pyridine (4.6 g, 58.198 mmol, 1.10 equiv). After stirring at 80 °C under nitrogen atmosphere overnight, the desired product can be detected by LCMS. The crude product was purified by reverse phase flash chromatography using the following conditions (column, C18 silica gel, mobile phase, MeCN / water (0.05% FA), 0% to 100% gradient in 30 minutes; detector, UV 254 nm) to provide intermediate 2 (6.4 g, 75.16%) as a red oil. LCMS (ESI) m / z: [M+H]+=294.

[1206] Step 2: Preparation of 2-methoxy-5-(piperidin-4-yl)pyrimidine (Intermediate 3)

[1207]

[1208] To a solution of intermediate 2 (6.4 g, 21.816 mmol, 1.00 equiv) in DCM (60.0 mL) and TFA (20.0 mL). After stirring at room temperature for 1 hour, the desired product can be detected by LCMS. The resulting mixture is concentrated under vacuum. This produces intermediate 3 (5.3 g, crude) as a green oil. LCMS (ESI) m / z: [M+H]+=194.

[1209] Step 3: Preparation of methyl 2-{3-[4-(2-methoxypyrimidin-5-yl)piperidin-1-yl]-1,2-oxazol-5-yl}-3-methylbutanoate (Intermediate 4)

[1210]

[1211] To a solution of intermediate 3 (5.3 g, crude) and methyl 3-methyl-2-{3-[(1,1,2,2,3,3,4,4,4-nonafluorobutanesulfonyl)oxy]-1,2-oxazol-5-yl}butanoate (21.3 g, 44.150 mmol, 2.00 equiv) in DMF (60 mL) was added DIEA (14.3 g, 110.375 mmol, 5.00 equiv). After stirring at 130 °C for 3 hours, the desired product can be detected by LCMS. The crude product was purified by reverse phase flash chromatography with the following conditions (column, C18 silica gel, mobile phase, MeCN / water (0.05% FA), 0% to 100% gradient in 30 minutes; detector, UV 254 nm) to provide intermediate 4 (736 mg, 8.90%) as a yellow oil. LCMS (ESI) m / z: [M+H]+=375.

[1212] Step 4: Preparation of methyl 2-{3-[4-(2-chloropyrimidin-5-yl)piperidin-1-yl]-1,2-oxazol-5-yl}-3-methylbutanoate (Intermediate 5)

[1213]

[1214] To a solution of POCl3 (904.1 mg, 5.898 mmol, 3.00 equiv) in DMF (8.0 mL) was added intermediate 4 (736.0 mg, 1.966 mmol, 1.00 equiv). After stirring at 100 °C for 2 hours, the desired product can be detected by LCMS. The crude product was purified by reverse phase flash chromatography using the following conditions (column, C18 silica gel, mobile phase, MeCN / water (0.05% FA), 0% to 100% gradient in 30 minutes; detector, UV 254 nm) to provide intermediate 5 (75.0 mg, 10.07%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=379.

[1215] Step 5: Preparation of methyl 2-{3-[4-(2-{4-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]piperidin-1-yl}pyrimidin-5-yl)piperidin-1-yl]-1,2-oxazol-5-yl}-3-methylbutanoate (Intermediate 6)

[1216]

[1217] To a solution of intermediate 5 (75.0 mg, 0.198 mmol, 1.00 equiv) and 2-[5-methyl-6-(piperidin-4-yl)thieno[2,3-c]pyridazin-3-yl]phenol (77.3 mg, 0.238 mmol, 1.20 equiv) in NMP (2 mL) was added K2CO3 (54.7 mg, 0.396 mmol, 2.00 equiv). After stirring at 100 °C for 3 h, the desired product could be detected by LCMS. The crude product was purified by reverse phase flash chromatography using the following conditions (column, C18 silica gel, mobile phase, MeCN / water (0.05% FA), 0% to 100% gradient in 30 min; detector, UV 254 nm) to provide intermediate 6 (58.0 mg, 43.87%) as a light yellow solid. LCMS (ESI) m / z: [M+H]+=668.

[1218] Step 6: Preparation of 2-{3-[4-(2-{4-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]piperidin-1-yl}pyrimidin-5-yl)piperidin-1-yl]-1,2-oxazol-5-yl}-3-methylbutanoic acid (Intermediate 7)

[1219]

[1220] To a solution of intermediate 6 (58.0 mg, 0.087 mmol, 1.00 equiv) and LiOH (6.3 mg, 0.261 mmol, 3.00 equiv) in MeOH (1.0 mL) and H2O (1.0 mL). After stirring at room temperature for 1 hour, the desired product can be detected by LCMS. The resulting mixture is extracted with EA (3 x 3.0 mL). The combined organic layers are washed with water (1 x 3.0 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate is concentrated under reduced pressure to provide intermediate 7 (48.0 mg, crude) as a white solid. LCMS (ESI) m / z: [M+H]+=654.

[1221] Step 7: Preparation of (2S,4R)-4-hydroxy-1-[(2R)-2-{3-[4-(2-{4-[(4Z)-4-[2-(2-hydroxyphenyl)-2-iminoethylidene]-3-methyl-5H-thiophen-2-yl]piperidin-1-yl}pyrimidin-5-yl)piperidin-1-yl]-1,2-oxazol-5-yl}-3-methylbutanoyl]-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (Intermediate 8)

[1222]

[1223] To a solution of intermediate 7 (45 mg, 0.069 mmol, 1 eq) and (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (22.8 mg, 0.069 mmol, 1.00 eq) in DMF (2.0 mg) was added DIEA (26.7 mg, 0.207 mmol, 3.00 eq) and HATU (31.4 mg, 0.083 mmol, 1.20 eq). After stirring at room temperature for 1 hour, the desired product could be detected by LCMS. The crude product was purified by reverse phase flash chromatography using the following conditions (column, C18 silica gel, mobile phase, MeCN / water (0.05% FA), 0% to 100% gradient in 30 minutes; detector, UV 254 nm) to provide intermediate 8 (48.0 mg, 72.93%) as a white solid. LCMS (ESI) m / z: [M+H]+=966.

[1224] Step 8: Preparation of (2S,4R)-4-hydroxy-1-[(2R)-2-{3-[4-(2-{4-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]piperidin-1-yl}pyrimidin-5-yl)piperidin-1-yl]-1,2-oxazol-5-yl}-3-methylbutyryl]-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide

[1225]

[1226] Intermediate 8 (48.0 mg) was purified by chiral HPLC under the following conditions: column, CHIRALPAK ID, 2*25 cm, 5 μm; mobile phase A: MtBE (10 Mm NH3-MeOH), mobile phase B: EtOH—HPLC; flow rate: 18 mL / min; gradient: 30% B to 30% B in 24 minutes; wavelength: 253 / 210 nm; RT1 (min): 11.24; RT2 (min): 18.89; sample solvent: MeOH:DCM=1:1—HPLC; injection volume: 1.5 mL; number of runs: 4, to provide the title compound (second peak) (4.6 mg, 29.11%) as an off-white solid.

[1227] 1H NMR (400MHz, DMSO-d6) δ12.78(s,1H),8.99(s,1H),8.71(s,1H),8.41(d,J=7 .7Hz,1H),8.35–8.29(m,2H),8.17(d,1H),7.49–7.41(m,2H),7.40–7.33(m, 3H),7.07–6.97(m,2H),6.12(s,1H),5.12(d,J=3.6Hz,1H),5.02–4.63(m,3H ),4.38(t,J=7.8Hz,1H),4.33–4.24(m,1H),3.83–3.66(m,3H),3.66–3.53(m ,2H),3.51(s,1H),3.47–3.39(m,1H),3.06(t,J=12.4Hz,2H),2.86(t,J=12. 0Hz,2H),2.70–2.56(m,1H),2.46(s,3H),2.37–2.12(m,2H),2.09–1.97(m,3 H),1.88–1.74(m,3H),1.76–1.53(m,5H),1.42(d,J=32.3,7.0Hz,3H),0.97( d,J=7.0Hz,3H),0.83(d,J=15.7,6.6Hz,3H).LCMS(ESI)m / z:[M+H]+=967.25.

[1228] (2S,4R)-4-Hydroxy-1-((R)-2-(3-((1s,4S)-4-(2-(4-(3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl)piperidin-1-yl)pyrimidin-5-yl)cyclohexyl)isoxazol-5-yl)-3-methylbutanoyl)-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[1229] and (2S,4R)-4-hydroxy-1-((R)-2-(3-((1r,4R)-4-(2-(4-(3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl)piperidin-1-yl)pyrimidin-5-yl)cyclohexyl)isoxazol-5-yl)-3-methylbutyryl)-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (Compound 64 and Compound 65)

[1230]

[1231] Step 1: Preparation of ethyl 4-(2-methoxypyrimidin-5-yl)cyclohex-3-ene-1-carboxylate (Intermediate 2)

[1232]

[1233] To a stirred solution of 5-bromo-2-methoxypyrimidine (3.37 g, 17.846 mmol, 1 eq) and ethyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylate (5 g, 17.846 mmol, 1.00 eq) in 1,4-dioxane (40 mL) and H2O (10 mL) was added Pd(dppf)Cl2.CH2Cl2 (1.45 g, 1.785 mmol, 0.1 eq) and K2CO3 (7.40 g, 53.538 mmol, 3 eq) at room temperature. The resulting mixture was stirred at 80 °C under nitrogen atmosphere for 2 hours. The mixture was cooled to room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (1 / 2) to afford intermediate 2 (3.8, 81.4%) as a yellow oil. LCMS (ESI) m / z: [M+H]+=263.

[1234] Step 2: Preparation of ethyl 4-(2-methoxypyrimidin-5-yl)cyclohexane-1-carboxylate (Intermediate 3)

[1235]

[1236] To a stirred solution of intermediate 2 (3.8 g, 14.50 mmol, 1 eq.) in THF (50 mL) was added Pd(OH)2 / C (2 g) at room temperature. The resulting mixture was stirred at room temperature under a hydrogen atmosphere for 16 hours. The resulting mixture was filtered and the filter cake was washed with THF. The filtrate was concentrated under reduced pressure. This produced intermediate 3 (3.1 g, 81.0%) as a grey oil. LCMS (ESI) m / z: [M+H]+=265.

[1237] Step 3: Preparation of [4-(2-methoxypyrimidin-5-yl)cyclohexyl]methanol (Intermediate 4)

[1238]

[1239] To a stirred solution of intermediate 3 (4 g, 15.133 mmol, 1 eq.) in THF (50 mL) was added LiAlH4 (0.57 g, 15.133 mmol, 1 eq.) at 0°C under nitrogen atmosphere. The resulting mixture was stirred at 0°C under nitrogen atmosphere for 1 hour. The reaction was quenched with water at 0°C. The resulting mixture was extracted with DCM (2 x 100 mL). The combined organic layers were washed with saturated brine (100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. This produced intermediate 4 (566 mg, 16.83%) as a yellow oil. LCMS (ESI) m / z: [M+H]+=223.

[1240] Step 4: Preparation of 4-(2-methoxypyrimidin-5-yl)cyclohexane-1-carbaldehyde (Intermediate 5)

[1241]

[1242] To a stirred solution of (COCl)2 (969.51 mg, 7.638 mmol, 3 eq.) in DCM (10 mL) was added DMSO (795.75 mg, 10.184 mmol, 4 eq.) at -78 °C under a nitrogen atmosphere. The resulting mixture was stirred at -78 °C under a nitrogen atmosphere for 30 minutes. Intermediate 4 (566 mg, 2.546 mmol, 1 eq.) was added to the above mixture at -78 °C under a nitrogen atmosphere. The resulting mixture was stirred for another 30 minutes at -78 °C under a nitrogen atmosphere. Et3N (1.29 g, 12.730 mmol, 5 eq.) was added to the above mixture at -78 °C under a nitrogen atmosphere. The resulting mixture was stirred for another 1 hour from -78 °C to room temperature under a nitrogen atmosphere. The reaction was quenched with water at room temperature and extracted with DCM (2 x 100 mL). The combined organic layers were washed with saturated brine (100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. This produced intermediate 5 (450 mg, 79.5%) as a yellow oil. LCMS (ESI) m / z: [M+H]+=221.

[1243] Step 5: Preparation of (Z)-N-{[4-(2-methoxypyrimidin-5-yl)cyclohexyl]methylidene}hydroxylamine (Intermediate 6)

[1244]

[1245] To a stirred solution of hydroxylamine hydrochloride (2.46 g, 35.412 mmol, 3 eq.) in MeOH (8 mL) and H2O (8 mL) was added Na2CO3 (3.75 g, 35.412 mmol, 3 eq.) at 0°C. To the above mixture was added intermediate 5 (2.6 g, 11.804 mmol, 1 eq.) at 0°C. The resulting mixture was stirred at room temperature for another 1 hour. The reaction was quenched with water at 0°C. The resulting mixture was extracted with EA (2 x 200 mL). The combined organic layers were washed with saturated brine (200 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. This produced intermediate 6 (2.8 g, crude) as a yellow oil. LCMS (ESI) m / z: [M+H]+=236.

[1246] Step 6: Preparation of (Z)-N-hydroxy-4-(2-methoxypyrimidin-5-yl)cyclohexane-1-carboximidoyl chloride (Intermediate 7)

[1247]

[1248] To a stirred solution of intermediate 6 (2.8 g, 11.900 mmol, 1 eq.) in EA (30 mL) was added NCS (1.91 g, 14.280 mmol, 1.2 eq.) at 0 °C. The resulting mixture was stirred at room temperature for 1 hour. The desired product can be detected by LCMS. The mixture was diluted with EtOAc (80 mL) and washed with water (80 mL x 2). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude intermediate 7 (3.2 g, crude) as a yellow solid. LCMS (ESI) m / z: [M+H]+=270.

[1249] Step 7: Preparation of methyl 2-{3-[4-(2-methoxypyrimidin-5-yl)cyclohexyl]-1,2-oxazol-5-yl}acetate (Intermediate 8)

[1250]

[1251] To a stirred solution of intermediate 7 (3.2 g, 11.864 mmol, 1 eq.) in EA (30 mL) was added NaHCO3 (2.99 g, 35.592 mmol, 3 eq.) at 0°C. To the above mixture was added but-3-ynoic acid methyl ester (4.66 g, 47.456 mmol, 4 eq.) at 0°C. The resulting mixture was stirred at room temperature for another 16 hours. The reaction was quenched with water. The resulting mixture was extracted with EA (2 x 200 mL). The combined organic layers were concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.1% FA), 0 to 100% gradient in 30 minutes; detector, UV 254 / 220 nm. This produced intermediate 8 (2.7 g, 68.68%) as a brown oil. LCMS (ESI) m / z: [M+H]+=332.

[1252] Step 8: Preparation of methyl 2-(3-(4-(2-methoxypyrimidin-5-yl)cyclohexyl)isoxazol-5-yl)-3-methylbutanoate (Intermediate 9)

[1253]

[1254] To a stirred solution of intermediate 8 (2.7 g, 8.148 mmol, 1 eq.) in THF (15 mL) was added t-BuOK (2.74 g, 24.444 mmol, 3 eq.) at 0 ° C. under a nitrogen atmosphere. The resulting mixture was stirred at 0 ° C. under a nitrogen atmosphere for 30 minutes. 2-iodopropane (2.77 g, 16.296 mmol, 2 eq.) was added to the above mixture at 0 ° C. under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for another 2 hours. The reaction was quenched with water at 0 ° C. The resulting mixture was extracted with DCM (2x200 mL). The combined organic layers were concentrated under reduced pressure. This produced 2-{3-[4-(2-methoxypyrimidine-5-yl)cyclohexyl]-1,2-oxazol-5-yl}-3-methylbutanoic acid (2 g, crude) as a brown oil. The product was dissolved in DCM (16 mL) and MeOH (4 mL), and TMSCHN2 (1.91 g, 16.692 mmol, 3 equiv.) was added at 0°C. The resulting mixture was stirred at room temperature for 1 hour. The reaction was quenched with water at 0°C. The resulting mixture was extracted with DCM (2 x 100 mL). The combined organic layers were concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.1% FA), 0 to 100% gradient in 30 minutes; detector, UV 254 / 220 nm. This produced intermediate 9 (795 mg, 38.26%) as a brown oil. LCMS (ESI) m / z: [M+H]+=374.

[1255] Step 9: Preparation of methyl 2-(3-(4-(2-chloropyrimidin-5-yl)cyclohexyl)isoxazol-5-yl)-3-methylbutanoate (Intermediate 10)

[1256]

[1257] To a stirred solution of intermediate 9 (795 mg, 2.129 mmol, 1 eq.) in DMF (5 mL) was added POCl3 (979.15 mg, 6.387 mmol, 3 eq.) at room temperature. The resulting mixture was stirred at 80°C for 16 hours. The mixture was allowed to cool to room temperature. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.1% FA), 0 to 100% gradient in 30 minutes; detector, UV 254 / 220 nm. This produced intermediate 10 (154 mg, 19.14%) as a brown oil. LCMS (ESI) m / z: [M+H]+=378.

[1258] Step 10: Preparation of methyl 2-{3-[4-(2-{4-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]piperidin-1-yl}pyrimidin-5-yl)cyclohexyl]-1,2-oxazol-5-yl}-3-methylbutanoate (Intermediate 11)

[1259]

[1260] To a stirred solution of intermediate 10 (100 mg, 0.265 mmol, 1 eq.) and 2-[5-methyl-6-(piperidin-4-yl)thieno[2,3-c]pyridazin-3-yl]phenol (103.35 mg, 0.318 mmol, 1.2 eq.) in DMSO (5 mL) was added DIEA (102.61 mg, 0.795 mmol, 3.0 eq.) at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 100 °C under nitrogen atmosphere for 2 hours. The desired product can be detected by LCMS. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeOH / water (0.1% FA), 0% to 100% gradient in 25 minutes; detector, UV 254 nm. This produced intermediate 11 (105 mg, 59.50%) as a light yellow solid. LCMS (ESI) m / z: [M+H]+=667.

[1261] Step 11: Preparation of methyl 2-(3-(4-(2-chloropyrimidin-5-yl)cyclohexyl)isoxazol-5-yl)-3-methylbutanoate (Intermediate 12)

[1262]

[1263] To a stirred solution of intermediate 11 (100 mg, 0.150 mmol, 1 eq.) in MeOH (5 mL) was added LiOH (17.96 mg, 0.750 mmol, 5.0 eq.) in H2O (2 mL). The resulting mixture was stirred at room temperature for 2 hours. The mixture was acidified to pH 5 with HCl (aq.). The resulting mixture was extracted with EtOAc (3x100 mL). The combined organic layers were washed with brine (50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. This produced intermediate 12 (85 mg, 86.83%) as a light yellow solid. LCMS (ESI) m / z: [M+H]+=653.

[1264] Step 12: Preparation of (2S,4R)-4-hydroxy-1-(2-{3-[4-(2-{4-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]piperidin-1-yl}pyrimidin-5-yl)cyclohexyl]-1,2-oxazol-5-yl}-3-methylbutanoyl)-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (Intermediate 13)

[1265]

[1266] To a stirred solution of intermediate 12 (85 mg, 0.130 mmol, 1 eq) and (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (61.99 mg, 0.195 mmol, 1.5 eq) in DMF (5 mL) was added PyBOP (101.64 mg, 0.195 mmol, 1.5 eq) and DIEA (50.49 mg, 0.390 mmol, 3.0 eq) at room temperature. The resulting mixture was stirred at room temperature for 4 hours. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeOH / water (0.1% TFA), 0% to 100% gradient in 30 minutes; detector, UV 254 nm. This resulted in intermediate 13 (106 mg, 84.25%) as an off-white solid. LCMS (ESI) m / z: [M+H]+ = 966.

[1267] Step 13: (2S,4R)-4-hydroxy-1-((R)-2-(3-((1s,4S)-4-(2-(4-(3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl)piperidin-1-yl)pyrimidin-5-yl)cyclohexyl)isoxazol-5-yl)-3-methylbutanoyl)-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide and

[1268] Preparation of (2S,4R)-4-hydroxy-1-((R)-2-(3-((1r,4R)-4-(2-(4-(3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl)piperidin-1-yl)pyrimidin-5-yl)cyclohexyl)isoxazol-5-yl)-3-methylbutyryl)-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[1269]

[1270] Intermediate 13 was purified by chiral preparative HPLC using the following conditions: column, CHIRAL ART Cellulose-SB, 2*25 cm, 5 um; mobile phase, MtBE (10 mM NH3-MeOH) and MeOH- (maintaining 20% ​​MeOH- within 12 minutes); detector, UV 254 nm to provide compound (2S,4R)-4-hydroxy-1-((R)-2-(3-((1s,4S)-4-(2-(4-(3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl)piperidin-1-yl)pyrimidin-5-yl)cyclohexyl)isoxazol-5-yl)-3-methylbutanoyl)-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (28.1 mg) as an off-white solid. 1H NMR (400MHz, DMSO-d6) δ12.79(s,1H),8.96(d,J=15.7Hz,1H),8.70(d,J=2.5Hz,1H ),8.43(d,J=7.7Hz,1H),8.24(d,J=5.2Hz,2H),8.20–8.13(m,1H),7.47–7.30(m,5 H),7.06–6.98(m,2H),6.35(s,1H),5.10(d,J=3.6Hz,1H),4.91(h,J=6.7Hz,1H),4 .80(t,J=14.7Hz,2H),4.38(t,J=7.9Hz,1H),4.32–4.27(m,1H),3.80–3.68(m,2H) ,3.65–3.54(m,1H),3.47(d,J=10.6Hz,1H),3.12–2.93(m,3H),2.59(d,J=12.9Hz, 1H),2.48(d,J=8.0Hz,3H),2.44(d,J=11.0Hz,3H),2.33–2.21(m,1H),2.11–1.96( m,5H),1.87–1.75(m,3H),1.73–1.69(m,2H),1.66–1.52(m,4H),1.41(d,J=16.1Hz ,3H),0.98(d,J=6.7Hz,3H),0.79(d,J=6.7Hz,3H).LCMS(ESI)m / z:[M+H]+=966.55.

[1271] and compound (2S,4R)-4-hydroxy-1-((R)-2-(3-((1r,4R)-4-(2-(4-(3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl)piperidin-1-yl)pyrimidin-5-yl)cyclohexyl)isoxazol-5-yl)-3-methylbutanoyl)-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (9.1 mg) as an off-white solid. 1HNMR(400MHz,DMSO-d6)δ12.79(s,1H),9.00(d,J=4.8Hz,1H),8.71(s,1H),8.44 (d,J=7.7Hz,1H),8.33(s,2H),8.17(d,J=7.8Hz,1H),7.51–7.42(m,2H),7.38(t,J =6.5Hz,3H),7.03(d,J=7.8Hz,2H),6.33(s,1H),5.12(d,J=3.6Hz,1H),4.93(q,J =6.8Hz,1H),4.84(d,J=13.2Hz,2H),4.38(t,J=7.9Hz,1H),4.30(s,1H),3.79–3.6 8(m,2H),3.66–3.56(m,1H),3.53–3.45(m,1H),3.05(t,J=12.5Hz,2H),2.74(s,1 H),2.46(d,J=2.6Hz,3H),2.32–2.14(m,2H),2.04(d,J=12.3Hz,5H),1.89–1.85(m ,2H),1.84–1.74(m,1H),1.67–1.51(m,7H),1.39(d,J=7.0Hz,3H),1.26–1.09(m, 2H), 0.98 (d, J = 6.3Hz, 3H), 0.79 (d, J = 6.7Hz, 3H). LCMS (ESI) m / z: [M+H]+ = 966.60.

[1272] Preparation of (2S,4R)-4-hydroxy-1-[(2R)-2-(3-{6-[3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl]-2-azaspiro[3.3]hept-2-yl}-1,2-oxazol-5-yl)-3-methylbutanoyl]-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (Compound 1)

[1273]

[1274] Step 1: Preparation of tert-butyl 6-[2-(3,6-dichloropyridazin-4-yl)ethynyl]-2-azaspiro[3.3]heptane-2-carboxylate (Intermediate 2)

[1275]

[1276] Pd(PPh3)2Cl2(0.26g, 0.364mmol, 0.1 eq) and TEA(1.10g, 10.914mmol, 3 eq) were added to a solution of 3,6-dichloro-4-iodopyridazine (1g, 3.638mmol, 1 eq) and 6-ethynyl-2-azaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester (0.81g, 3.638mmol, 1 eq) in toluene (10mL), and the resulting solution was stirred at 60°C for 6 hours. The mixture was diluted with EtOAc (400mL) and washed with water (400mLx 3). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography with an elution gradient of 0 to 43% PE / EA to obtain intermediate 2 (1.11g, 82.85%) as a brown solid. LCMS (ESI) m / z: [M+H]+=368.

[1277] Step 2: Preparation of tert-butyl 6-{3-chlorothieno[2,3-c]pyridazin-6-yl}-2-azaspiro[3.3]heptane-2-carboxylate (Intermediate 3)

[1278]

[1279] To a solution of intermediate 2 (1.1 g, 2.987 mmol, 1 eq.) in NMP (10 mL) was added NaSH (0.17 g, 2.987 mmol, 1 eq.) and the resulting solution was stirred at 100 °C for 6 h. Without additional workup, the crude reaction solution was purified by flash C18 chromatography with an elution gradient of 0 to 46% ACN / H2O to afford intermediate 3 (422 mg, 38.61%) as a brown solid. LCMS (ESI) m / z: [M+H]+=366.

[1280] Step 3: Preparation of tert-butyl 6-[3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl]-2-azaspiro[3.3]heptane-2-carboxylate (Intermediate 4)

[1281]

[1282] To a solution of tert-butyl intermediate 3 (400 mg, 1.093 mmol, 1 eq) and 2-hydroxyphenylboronic acid (226.19 mg, 1.639 mmol, 1.5 eq) in 1,4-dioxane (4 mL) and H2O (1 mL) was added XPhos Pd G3 (92.54 mg, 0.109 mmol, 0.1 eq) and Cs2CO3 (712.41 mg, 2.186 mmol, 2 eq) and the resulting solution was stirred at 80 °C for 3 hours. The mixture was diluted with EtOAc (300 mL) and washed with water (300 mL x2). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product. The crude product was purified by flash C18 chromatography with an elution gradient of 0 to 41% ACN / H2O to give intermediate 4 (347 mg, 74.94%) as a brown solid. LCMS (ESI) m / z: [M+H]+=424.

[1283] Step 4: Preparation of 2-(6-{2-azaspiro[3.3]hept-6-yl}thieno[2,3-c]pyridazin-3-yl)phenol (Intermediate 5)

[1284]

[1285] To a solution of intermediate 4 (347 mg, 0.819 mmol, 1 eq.) in DCM (3 mL) was added TFA (1 mL) and the resulting solution was stirred at 25° C. for 2 hours. The resulting mixture was concentrated under reduced pressure to give intermediate 5 (308 mg, crude) as a yellow solid. LCMS (ESI) m / z: [M+H]+=324.

[1286] Step 5: Preparation of methyl 2-(3-{6-[3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl]-2-azaspiro[3.3]hept-2-yl}-1,2-oxazol-5-yl)-3-methylbutanoate (Intermediate 6)

[1287]

[1288] To a solution of intermediate 5 (250 mg, 0.773 mmol, 1 eq) and methyl 3-methyl-2-{3-[(1,1,2,2,3,3,4,4,4-nonafluorobutanesulfonyl)oxy]-1,2-oxazol-5-yl}butanoate (372.04 mg, 0.773 mmol, 1 eq) in DMSO (3 mL) was added DIEA (299.73 mg, 2.319 mmol, 3 eq) and the resulting solution was stirred at 100 °C for 5 hours. The crude reaction solution was purified by flash C18 chromatography with an elution gradient of 0 to 45% ACN / H2O to afford intermediate 6 (203 mg, 52.04%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=505.

[1289] Step 6: Preparation of 2-(3-{6-[3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl]-2-azaspiro[3.3]hept-2-yl}-1,2-oxazol-5-yl)-3-methylbutanoic acid (Intermediate 7)

[1290]

[1291] To a solution of intermediate 6 (200 mg, 0.396 mmol, 1 eq.) in MeOH (2 mL) and H2O (0.5 mL) was added LiOH (47.46 mg, 1.980 mmol, 5 eq.), and the resulting solution was stirred at 25 °C for 2 hours. The mixture was acidified to pH 5-6 with HCl (1 M in H2O). The resulting mixture was diluted with water (100 mL). The resulting mixture was extracted with EA (100 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give intermediate 7 (297 mg, crude) as a brown solid. LCMS (ESI) m / z: [M+H]+=491.

[1292] Step 7: Preparation of (2S,4R)-4-hydroxy-1-[2-(3-{6-[3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl]-2-azaspiro[3.3]hept-2-yl}-1,2-oxazol-5-yl)-3-methylbutanoyl]-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (Intermediate 8)

[1293]

[1294] To a solution of Intermediate 7 (200 mg, 0.408 mmol, 1 eq.) and (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (270.24 mg, 0.816 mmol, 2 eq.) in DMF (3 mL) was added PyBOP (424.31 mg, 0.816 mmol, 2 eq.) and DIEA (263.46 mg, 2.040 mmol, 5 eq.) and the resulting solution was stirred at 25 °C for 2 h. Without additional work-up, the crude reaction solution was purified by flash C18 chromatography with an elution gradient of 0 to 52% ACN / H2O to afford Intermediate 8 (184 mg, 56.2%). LCMS (ESI) m / z: [M+H]+=804.

[1295] Step 7: Preparation of (2S,4R)-4-hydroxy-1-[(2R)-2-(3-{6-[3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl]-2-azaspiro[3.3]hept-2-yl}-1,2-oxazol-5-yl)-3-methylbutanoyl]-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide

[1296]

[1297] Intermediate 8 (184 mg) was purified by chiral-HPLC (column: CHIRALPAKID, 2*25 cm, 5 μm; mobile phase A: MtBE (10 mM NH3-MeOH), mobile phase B: MeOH--HPLC; flow rate: 20 mL / min; gradient: 30% B to 30% B in 17 minutes; wavelength: 208 / 272 nm; RT1 (min): 6.345; RT2 (min): 8.7275; sample solvent: MeOH--HPLC; injection volume: 0.7 mL; number of runs: 7) to give the title compound (30.5 mg, 16.57%) as an off-white solid (second peak). 1H NMR (400 MHz, methanol-d4) δ8.87 (s, 1H), 8.58 (s, 1H), 7.95 (d, J = 8.1 Hz, 1H), 7.61–7.31 (m, 5H), 7.27 (s, 1H), 7.01 (d, J = 8.0 Hz, 2H), 5.88 (s, 1H), 5.03 (q, J = 7.3 Hz, 1H), 4.51 (t, J = 8.2 Hz, 1H), 4.44 (s, 1H), 4.11 (s, 2H), 3.93 (s, 2H), 4.03–3.89 (m ,2H),3.62(t,J=12.1Hz,2H),2.83(s,2H),2.57(t,J=10.2Hz,2H),2.48(s,3H),2.41–2.29(m,1H),2.23–2.08(m,1H), 2.03–1.81(m,1H),1.52(d,J=7.1Hz,3H),1.05(d,J=6.5Hz,3H),0.89(d,J=7.2Hz,3H).LCMS(ESI)m / z:[M+H]+=804.20.

[1298] The compounds in Table 2 were prepared using a procedure similar to that used above to prepare (2S,4R)-4-hydroxy-1-[(2R)-2-(3-{6-[3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl]-2-azaspiro[3.3]hept-2-yl}-1,2-oxazol-5-yl)-3-methylbutanoyl]-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (Compound 1).

[1299] Table 2.

[1300]

[1301]

[1302]

[1303]

[1304]

[1305]

[1306]

[1307]

[1308] Preparation of (2S,4R)-4-hydroxy-1-((R)-2-(3-(2-(3-(2-hydroxyphenyl)thieno[3,2-c]pyridazin-6-yl)-7-azaspiro[3.5]non-7-yl)isoxazol-5-yl)-3-methylbutyryl)-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (Compound 12)

[1309]

[1310] Step 1: Preparation of 4-bromo-6-chloro-3-iodopyridazine (Intermediate 2)

[1311]

[1312] To a stirred solution of 4-bromo-6-chloropyridazin-3-amine (5 g, 23.988 mmol, 1 eq.) and CuI (5.48 g, 28.786 mmol, 1.2 eq.) in THF (60 mL) and CH2I2 (7.71 g, 28.786 mmol, 1.2 eq.) was added t-BuNO2 (2.47 g, 23.988 mmol, 1 eq.) after stirring the resulting solution at 25°C for 0.5 h. The crude product was purified by silica gel column chromatography with an elution gradient of 0 to 80% EA / PE to afford intermediate 2 (2.9 g, 37.86%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=319.

[1313] Step 2: Preparation of tert-butyl 2-((4-bromo-6-chloropyridazin-3-yl)ethynyl)-7-azaspiro[3.5]nonane-7-carboxylate (Intermediate 3)

[1314]

[1315] To a solution of intermediate 2 (2 g, 6.263 mmol, 1 eq.) and tert-butyl 2-ethynyl-7-azaspiro[3.5]nonane-7-carboxylate (1.56 g, 6.263 mmol, 1 eq.) and Pd(PPh3)2Cl2 (0.88 g, 1.253 mmol, 0.2 eq.) and CuI (0.24 g, 1.253 mmol, 0.2 eq.) in methylbenzene (15 mL, 0.011 mmol) and TEA (1.90 g, 18.789 mmol, 3 eq.) was stirred at 25°C for 2 hours. The crude product was purified by silica gel column chromatography with an elution gradient of 0 to 17% EA / PE to afford intermediate 3 (1.6 g, 57.96%) as a yellow oil. LCMS (ESI) m / z: [M+H]+ = 441.

[1316] Step 3: Preparation of tert-butyl 2-(3-chlorothieno[3,2-c]pyridazin-6-yl)-7-azaspiro[3.5]nonane-7-carboxylate (Intermediate 4)

[1317]

[1318] To a solution of intermediate 3 (1.6 g, 3.630 mmol, 1 eq.) and NaSH (203.50 mg, 3.630 mmol, 1 eq.) in NMP (10 mL, 51.850 mmol), the resulting solution was stirred at 100 °C for 2 hours. The resulting mixture was diluted with EA (400 mL) and washed with water (3 x 400 mL). The organic layer was dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with an elution gradient of 0 to 34% EA / PE to obtain intermediate 4 (890 mg, 62.24%) as an orange solid. LCMS (ESI) m / z: [M+H]+=394.

[1319] Step 4: Preparation of tert-butyl 2-(3-(2-hydroxyphenyl)thieno[3,2-c]pyridazin-6-yl)-7-azaspiro[3.5]nonane-7-carboxylate (Intermediate 5)

[1320]

[1321] To a solution of intermediate 4 (890 mg, 2.259 mmol, 1 eq.) and 2-hydroxyphenylboronic acid (467.43 mg, 3.388 mmol, 1.5 eq.), Cs2CO3 (1.47 g, 4.518 mmol, 2 eq.), XPhos Pd G3 (191.24 mg, 0.226 mmol, 0.1 eq.) in 1,4-dioxane (4 mL) and H2O (1 mL), the resulting solution was stirred at 80 °C for 2 hours. The resulting mixture was diluted with EA (400 mL) and washed with water (3 x 400 mL). The organic layer was dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with an elution gradient of 0 to 15% EA / PE to give intermediate 5 (571 mg, 55.97%) as a brown oil. LCMS (ESI) m / z: [M+H]+=452.

[1322] Step 5: Preparation of 2-(6-(7-azaspiro[3.5]nonan-2-yl)thieno[3,2-c]pyridazin-3-yl)phenol (Intermediate 6)

[1323]

[1324] To a solution of intermediate 5 (571 mg, 1.264 mmol, 1 eq) in TFA (1 mL) and DCM (3 mL), the resulting solution was stirred at 25 °C for 2 hours. The reaction was concentrated under reduced pressure to give intermediate 6 (667 mg, crude) as a brown oil, which was used directly in the next step without further purification. LCMS (ESI) m / z: [M+H]+=352

[1325] Step 6: Preparation of methyl 3-methyl-2-{3-[2-(3-{2-[(1,1,2,2,3,3,4,4,4-nonafluorobutanesulfonyl)oxy]phenyl}thieno[3,2-c]pyridazin-6-yl)-7-azaspiro[3.5]nonan-7-yl]-1,2-oxazol-5-yl}butanoate (Intermediate 7a)

[1326]

[1327] To a stirred solution of intermediate 6 (300 mg, 0.854 mmol, 1 eq) and methyl 3-methyl-2-{3-[(1,1,2,2,3,3,4,4,4-nonafluorobutanesulfonyl)oxy]-1,2-oxazol-5-yl}butanoate (410.81 mg, 0.854 mmol, 1.0 eq) in DMSO (5 mL) was added DIEA (330.96 mg, 2.562 mmol, 3.0 eq) at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 100 °C under nitrogen atmosphere for 2 hours. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.1% TFA), 0% to 100% gradient in 10 minutes; detector, UV 254 nm. This produced intermediate 7a (181 mg, 26.03%) as a tan solid. LCMS (ESI) m / z: [M+H]+=815.

[1328] Step 7: Preparation of 2-(3-(2-(3-(2-hydroxyphenyl)thieno[3,2-c]pyridazin-6-yl)-7-azaspiro[3.5]nonan-7-yl)isoxazol-5-yl)-3-methylbutanoic acid (Intermediate 8)

[1329]

[1330] To a solution of intermediate 7 (170 mg, 0.209 mmol, 1 eq.) and NaOH (83.46 mg, 2.090 mmol, 10 eq.) in MeOH (4 mL) and H2O (1 mL), the resulting solution was stirred at 25 °C for 4 hours. The mixture was acidified to pH 6 with concentrated HCl. The resulting mixture was extracted with EtOAc (3 x 200 mL). The combined organic layers were washed with brine (3 x 10 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to give intermediate 8 (155 mg, crude) as a brown solid. LCMS (ESI) m / z: [M+H]+ = 519.

[1331] Step 8: Preparation of (2S,4R)-4-hydroxy-1-(2-(3-(2-(3-(2-hydroxyphenyl)thieno[3,2-c]pyridazin-6-yl)-7-azaspiro[3.5]non-7-yl)isoxazol-5-yl)-3-methylbutanoyl)-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (Intermediate 9)

[1332]

[1333] To a solution of intermediate 8 (155 mg, 0.299 mmol, 1 eq) and (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (99.05 mg, 0.299 mmol, 1 eq) and PyBOP (311.06 mg, 0.598 mmol, 2 eq) in DMF (2 mL) and DIEA (193.14 mg, 1.495 mmol, 5 eq) the resulting solution was stirred at 25 °C for 2 h. Without additional work-up, the crude reaction solution was purified by preparative HPLC (column: XBridge Prep OBD C18 column, 30*150 mm, 5 μm; mobile phase A: water (10 mmol / L NH4HCO3 + 0.1% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 45% B to 70% B, 70% B in 7 minutes; wavelength: 254 / 220 nm; RT1 (min): 7.55; to obtain intermediate 9 (30 mg, 34.72%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=832.

[1334] Step 9: Preparation of (2S,4R)-4-hydroxy-1-((R)-2-(3-(2-(3-(2-hydroxyphenyl)thieno[3,2-c]pyridazin-6-yl)-7-azaspiro[3.5]non-7-yl)isoxazol-5-yl)-3-methylbutanoyl)-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[1335]

[1336] Intermediate 9 was purified by chiral preparative HPLC under the following conditions: column: CHIRAL ART Cellulose-SB, 2*25 cm, 5 μm; mobile phase A: MtBE (10 mM NH3-MeOH), mobile phase B: MeOH--HPLC; flow rate: 20 mL / min; gradient: 20% B to 20% B in 13 minutes; wavelength: 270 / 212 nm; RT1 (min): 7.8; RT2 (min): 10.4; sample solvent: MeOH:DCM=1:2; injection volume: 0.5 mL; number of runs: 4. This produced the title compound (second peak) as a white solid (11.8 mg, 4.51%). (1HNMR (300MHz, methanol-d4) δ8.92(d,J=15.4Hz,2H),7.99(d,J=8.0Hz,1H),7.56(s,1H),7.52–7.37(m,4H),7.04(d,J=7.8Hz,2H),6.11(d,J=16.1Hz ,1H),5.12–5.02(m,1H),4.54(t,J=8.1Hz,1H),4.46(s,1H),4.07–4.42 (m,1H),4.05(dd,J=10.8,4.1Hz,1H),3.92–3.83(m,1H),3.64(dd,J=10. 4,6.1Hz,2H),3.45(s,2H),3.35(m,2H),3.18(d,J=6.3Hz,2H),2.56(t, J=10.3Hz,2H),2.50(s,3H),2.39(s,1H),2.20(t,J=10.3Hz,3H),2.06–1 .93(m,1H),1.91(s,2H),1.75(s,2H),1.58(dd,J=20.1,7.0Hz,3H),1.08(d,J=6.5Hz,3H),0.92(d,J=6.7Hz,3H).LCMS(ESI)m / z:[M+H]+=832.35

[1337] Preparation of (2S,4R)-4-hydroxy-1-((R)-2-(3-(6-(4-(3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl)piperidin-1-yl)-5-methylpyridin-3-yl)isoxazol-5-yl)-3-methylbutyryl)-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (Compound 2)

[1338]

[1339] Step 1: Preparation of methyl 2-(3-(6-(4-(3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl)piperidin-1-yl)-5-methylpyridin-3-yl)isoxazol-5-yl)-3-methylbutanoate

[1340]

[1341] To a stirred solution of intermediate 3 (200 mg, 0.642 mmol, 1 eq.) and intermediate 7 (187.74 mg, 0.642 mmol, 1 eq.) in DMSO (4 mL) was added DIEA (249.04 mg, 1.926 mmol, 3 eq.) dropwise at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 110 ° C. under nitrogen atmosphere for 36 hours. The desired product can be detected by LCMS. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (10 mmol / L NH4HCO3), 0% to 100% gradient in 30 minutes; detector, UV254 nm. The resulting mixture was concentrated under reduced pressure. This resulted in methyl 2-(3-(6-(4-(3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl)piperidin-1-yl)-5-methylpyridin-3-yl)isoxazol-5-yl)-3-methylbutanoate (50 mg) as a yellow solid. LCMS (ESI) m / z: [M+H]+=384.

[1342] Step 2: Preparation of 2-(3-(6-(4-(3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl)piperidin-1-yl)-5-methylpyridin-3-yl)isoxazol-5-yl)-3-methylbutanoic acid

[1343]

[1344] A solution of methyl 2-(3-(6-(4-(3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl)piperidin-1-yl)-5-methylpyridin-3-yl)isoxazol-5-yl)-3-methylbutanoate (50 mg, 0.086 mmol, 1 eq) in MeOH (2 mL) was treated with LiOH.HO (20.52 mg, 0.860 mmol, 10 eq) at room temperature, and then HO (1 mL) was added dropwise at room temperature. The resulting mixture was stirred at room temperature for 1 hour. The mixture was acidified to pH 5 with HCl (aq). The precipitated solid was collected by filtration and washed with HO (3 x 10 mL). This resulted in 2-(3-(6-(4-(3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl)piperidin-1-yl)-5-methylpyridin-3-yl)isoxazol-5-yl)-3-methylbutanoic acid (47 mg) as a yellow solid. LCMS (ESI) m / z: [M+H]+=570.

[1345] Step 3: Preparation of (2S,4R)-4-hydroxy-1-(2-(3-(6-(4-(3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl)piperidin-1-yl)-5-methylpyridin-3-yl)isoxazol-5-yl)-3-methylbutanoyl)-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[1346]

[1347] To a stirred solution of 2-(3-(6-(4-(3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl)piperidin-1-yl)-5-methylpyridin-3-yl)isoxazol-5-yl)-3-methylbutanoic acid (40 mg, 0.070 mmol, 1 eq) and intermediate 8 (27.93 mg, 0.084 mmol, 1.2 eq) in DMF (0.5 mL) was added PyBOP (73.08 mg, 0.140 mmol, 2 eq) and DIEA (27.23 mg, 0.210 mmol, 3 eq) portionwise at room temperature. The resulting mixture was stirred at room temperature for 1.5 hours. The desired product could be detected by LCMS. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (10 mmol / L NH4HCO3), gradient from 0% to 100% in 30 minutes; detector, UV 254 nm. The resulting mixture was concentrated under reduced pressure. This produced (2S,4R)-4-hydroxy-1-(2-(3-(6-(4-(3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl)piperidin-1-yl)-5-methylpyridin-3-yl)isoxazol-5-yl)-3-methylbutanoyl)-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (35 mg) as a light yellow solid. LCMS (ESI) m / z: [M+H]+=883.

[1348] Step 6: Preparation of (2S,4R)-4-hydroxy-1-((R)-2-(3-(6-(4-(3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl)piperidin-1-yl)-5-methylpyridin-3-yl)isoxazol-5-yl)-3-methylbutanoyl)-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[1349]

[1350] The above product was purified by chiral preparative HPLC under the following conditions: column, CHIRALPAK ID, 2*25 cm, 5 um; mobile phase, MtBE:DCM=2:1 (10 mM / L) and MeOH- (maintaining 50% MeOH- within 12 minutes); detector, UV 254. This produced the title compound (second peak) as an off-white solid (14.3 mg). 1HNMR (300MHz, DMSO-d6) δ12.38(s,1H),8.99(s,1H),8.74(s,1H),8.60(d,J=2.3Hz,1H),8.46(t,J=8.1Hz,1H),8.07–7.96(m,2H),7.48 –7.34(m,6H),7.09–6.97(m,2H),6.87(d,J=39.5Hz,1H),5.13(s,1H),5.02–4.88(m,1H),4.40(t,J=7.9Hz,1H),4.31(s,1H),3.86(d,J= 9.6Hz,1H),3.78–3.68(m,3H),3.53(d,J=7.6Hz,2H),3.00(t,J=12.1Hz,2H),2.47(s,3H),2.37–2.33(m,3H),2.22(d,J=12.5Hz,3H),2. 11–1.89(m,3H),1.82(d,J=8.7Hz,1H),1.54–1.36(m,3H),1.02(d,J=6.4Hz,3H),0.84(d,J=6.8Hz,3H).LCMS(ESI)m / z:[M+H]+=883.25.

[1351] The compounds in Table 3 were prepared using a procedure analogous to that used above to prepare (2S,4R)-4-hydroxy-1-((R)-2-(3-(6-(4-(3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl)piperidin-1-yl)-5-methylpyridin-3-yl)isoxazol-5-yl)-3-methylbutanoyl)-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (Compound 2) using the appropriate fluoropyridine and 2-(5-methyl-6-(piperidin-4-yl)thieno[2,3-c]pyridazin-3-yl)phenol.

[1352] Table 3.

[1353]

[1354]

[1355]

[1356]

[1357]

[1358]

[1359]

[1360] Preparation of (2S,4R)-4-hydroxy-1-[(2R)-2-[3-(4-{4-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]piperidin-1-yl}-3-methylphenyl)-1,2-oxazol-5-yl]-3-methylbutyryl]-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (Compound 3)

[1361]

[1362] Step 1: Preparation of ethyl (E)-N-[(4-bromo-3-methylphenyl)methylidene]hydroxylamine (Intermediate 2)

[1363]

[1364] A solution of 4-bromo-3-methylbenzaldehyde (4 g, 20.096 mmol, 1 eq.) and hydroxylamine hydrochloride (2.79 g, 40.192 mmol, 2.0 eq.) in MeOH (20 mL), H2O (20 mL) was stirred at room temperature for 4 hours. The resulting mixture was diluted with water (100 mL). The resulting mixture was extracted with CH2Cl2 (3 x 150 mL). The combined organic layers were washed with brine (2 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to provide intermediate 2 (4.5 g, 104.61%) as a white solid. The crude product was used directly in the next step without further purification. LCMS (ESI) m / z [M+H]+ = 214.

[1365] Step 2: Preparation of (Z)-4-bromo-N-hydroxy-3-methylbenzimidoyl chloride (Intermediate 3)

[1366]

[1367] A solution of intermediate 2 (4.4 g, 20.555 mmol, 1 eq.) and NCS (4.12 g, 30.832 mmol, 1.5 eq.) in EtOAc (40 mL) was stirred at room temperature for 12 hours. The resulting mixture was diluted with water (300 mL). The resulting mixture was extracted with EtOAc (2 x 300 mL). The combined organic layers were washed with brine (1 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to provide intermediate 3 (3.0 g, 58.73%) as a yellow solid. The crude product was used directly in the next step without further purification. LCMS (ESI) m / z[M+H]+=248.

[1368] Step 3: Preparation of methyl 2-[3-(4-bromo-3-methylphenyl)-1,2-oxazol-5-yl]acetate (Intermediate 4)

[1369]

[1370] A solution of Intermediate 3 (3.0 g, 12.072 mmol, 1 eq.) and but-3-ynoic acid methyl ester (1.18 g, 12.072 mmol, 1.0 eq.) in EtOAc (2 mL) was stirred at room temperature for 12 hours. The residue was purified by silica gel column chromatography eluting with PE / EA (1:1) to provide Intermediate 4 (3.0 g, 80.12%) as a yellow solid. LCMS (ESI) m / z [M+H] + = 310.

[1371] Step 4: Preparation of ethyl 2-[3-(4-bromo-3-methylphenyl)-1,2-oxazol-5-yl]-3-methylbutanoate (Intermediate 5)

[1372]

[1373] A solution of intermediate 4 (3.0 g, 9.673 mmol, 1 eq.) and 2-iodopropane (3.29 g, 19.346 mmol, 2.0 eq.) in THF (20 mL) was stirred at 60 °C for 4 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (7:3) to provide intermediate 5 (1.7 g, 49.90%) as a yellow oil. LCMS (ESI) m / z [M+H] + = 352.

[1374] Step 5: Preparation of methyl 2-[3-(4-{4-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]piperidin-1-yl}-3-methylphenyl)-1,2-oxazol-5-yl]-3-methylbutanoate (Intermediate 6)

[1375]

[1376] To a solution of intermediate 5 (200 mg, 0.568 mmol, 1 eq) and 2-[5-methyl-6-(piperidin-4-yl)thieno[2,3-c]pyridazin-3-yl]phenol (221.74 mg, 0.682 mmol, 1.2 eq) in dioxane (1 mL) was added Cs2CO3 (555.01 mg, 1.704 mmol, 3.0 eq), Pd2(dba)3 (311.98 mg, 0.341 mmol, 0.6 eq) and BINAP (106.07 mg, 0.170 mmol, 0.3 eq). After stirring at 100 °C under nitrogen atmosphere for 6 hours, the resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (7:3) to provide intermediate 6 (80 mg, 23.61%) as a yellow solid. LCMS (ESI) m / z [M+H] + = 597.

[1377] Step 6: Preparation of 2-[3-(4-{4-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]piperidin-1-yl}-3-methylphenyl)-1,2-oxazol-5-yl]-3-methylbutanoic acid (Intermediate 7)

[1378]

[1379] A solution of intermediate 6 (80 mg, 0.134 mmol, 1 eq.) and LiOH (64.21 mg, 2.680 mmol, 20 eq.) in MeOH (4 mL) was stirred at 60 °C for 1 hour. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.1% FA), 0% to 60% gradient in 40 minutes; detector, UV 254 nm. This produced intermediate 7 (45 mg, 57.60%) as a white solid. LCMS (ESI) m / z [M+H] + = 583.

[1380] Step 7: Preparation of tert-butyl (2S, 4R)-4-hydroxy-1-{2-[3-(4-{4-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]piperidin-1-yl}-3-methylphenyl)-1,2-oxazol-5-yl]-3-methylbutanoyl}-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (Intermediate 8)

[1381]

[1382] A solution of intermediate 7 (45 mg, 0.077 mmol, 1 eq.) and (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (30.71 mg, 0.092 mmol, 1.2 eq.), PyBOP (80.37 mg, 0.154 mmol, 2.0 eq.), DIEA (29.94 mg, 0.231 mmol, 3.0 eq.) in DMF (1 mL) was stirred at room temperature for 1 hour. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.1% FA), gradient 0% to 80% in 40 minutes; detector, UV 254 nm. This gave intermediate 8 (34 mg, 49.13%) as a white solid. LCMS (ESI) m / z [M+H] + = 896.

[1383] Step 8: Preparation of (2S,4R)-4-hydroxy-1-[(2R)-2-[3-(4-{4-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]piperidin-1-yl}-3-methylphenyl)-1,2-oxazol-5-yl]-3-methylbutyryl]-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide

[1384]

[1385] Intermediate 8 (34 mg) was purified by chiral preparative HPLC using the following conditions (column: CHIRALPAK ID-3, 4.6*50 mm, 3 um; mobile phase A: MtBE (0.1% DEA): MeOH = 70:30; flow rate: 1 mL / min; gradient: 0% B to 0% B; injection volume: 5 ul mL) to provide the title compound (second peak) as a white solid (11.3 mg, 33.24%). NMR(400MHz,DMSO-d6)δ12.81(d,J=3.3Hz,1H),8.99(s,1H),8.72(s,1H),8 .44(d,J=7.6Hz,1H),8.22–8.15(m,1H),7.73–7.64(m,2H),7.48–7.42(m,2 H),7.39(t,J=6.8Hz,3H),7.16(dd,J=8.5,3.9Hz,1H),7.07–6.99(m,2H),6 .86(s,1H),5.11(d,J=3.7Hz,1H),4.93(p,J=7.2Hz,1H),4.40(t,J=7.9Hz, 1H),4.30(brs,1H),3.84(d,J=9.7Hz,1H),3.76(dd,J=10.7,4.3Hz,1H),3. 68–3.38(m,3H),3.33–3.32(m,3H),2.89(t,J=11.6Hz,2H),2.63–2.53(m,1 H),2.45(s,3H),2.37(s,3H),2.18–1.72(m,7H),1.39(d,J=7.0Hz,3H),1.0 5–0.98(m,3H),0.85(dd,J=11.6,6.6Hz,3H).LCMS(ESI)m / z[M+H]+=896.25.

[1386] The compounds in Table 4 were prepared using a procedure analogous to that used above to prepare (2S,4R)-4-hydroxy-1-[(2R)-2-[3-(4-{4-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]piperidin-1-yl}-3-methylphenyl)-1,2-oxazol-5-yl]-3-methylbutanoyl]-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (Compound 3) using the appropriate aryl bromide and 2-(5-methyl-6-(piperidin-4-yl)thieno[2,3-c]pyridazin-3-yl)phenol.

[1387] Table 4.

[1388]

[1389]

[1390]

[1391] Preparation of (2S,4R)-4-hydroxy-1-[(2R)-2-(3-{2-[(3R)-3-[3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl]pyrrolidin-1-yl]pyrimidin-5-yl}-1,2-oxazol-5-yl)-3-methylbutyryl]-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (Compound 5)

[1392]

[1393] Step 1: Preparation of methyl 2-(3-{2-[(3R)-3-[3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl]pyrrolidin-1-yl]pyrimidin-5-yl}-1,2-oxazol-5-yl)-3-methylbutanoate (Intermediate 2)

[1394]

[1395] A mixture of 2-{6-[(3S)-pyrrolidin-3-yl]thieno[2,3-c]pyridazin-3-yl}phenol (400 mg, 1.345 mmol, 1 eq), methyl 2-[3-(2-chloropyrimidin-5-yl)-1,2-oxazol-5-yl]-3-methylbutanoate (397.77 mg, 1.345 mmol, 1 eq) and DIEA (869.24 mg, 6.725 mmol, 5 eq) in DMSO (4 mL) was stirred at 100° C. under nitrogen atmosphere for 3 hours. The desired product can be detected by LCMS. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeOH / water (0.1% FA), gradient from 0% to 100% in 30 minutes; detector, UV 254 nm to provide intermediate 2 (130 mg, 17.36%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=557.

[1396] Step 2: Preparation of 2-(3-{2-[(3R)-3-[3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl]pyrrolidin-1-yl]pyrimidin-5-yl}-1,2-oxazol-5-yl)-3-methylbutanoic acid (Intermediate 3)

[1397]

[1398] A mixture of intermediate 2 (130 mg, 0.234 mmol, 1 eq.) and LiOH (16.78 mg, 0.702 mmol, 3 eq.) in MeOH (2 mL), THF (2 mL) and H2O (1 mL) was stirred at room temperature for 2 hours. The desired product can be detected by LCMS. The mixture was acidified to pH 6 with concentrated HCl. The resulting mixture was concentrated under reduced pressure to provide intermediate 3 (142 mg, crude) as a yellow solid. LCMS (ESI) m / z [M+H]+ = 543.

[1399] Step 3: Preparation of (2S,4R)-4-hydroxy-1-[2-(3-{2-[(3R)-3-[3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl]pyrrolidin-1-yl]pyrimidin-5-yl}-1,2-oxazol-5-yl)-3-methylbutanoyl]-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (Intermediate 4)

[1400]

[1401] A mixture of Intermediate 3 (142 mg, 0.262 mmol, 1 eq), (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (86.73 mg, 0.262 mmol, 1 eq), PyBOP (204.28 mg, 0.393 mmol, 1.5 eq) and DIEA (169.12 mg, 1.310 mmol, 5 eq) in DMF (3 mL) was stirred at room temperature for 2 hours. The desired product can be detected by LCMS. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (10 mmol / L NH4HCO3), 0% to 100% gradient in 30 minutes; detector, UV 254 nm to provide intermediate 4 (100 mg, 44.64%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=856.

[1402] Step 4: Preparation of (2S,4R)-4-hydroxy-1-[(2R)-2-(3-{2-[(3R)-3-[3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl]pyrrolidin-1-yl]pyrimidin-5-yl}-1,2-oxazol-5-yl)-3-methylbutanoyl]-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide

[1403]

[1404] The product intermediate 4 was purified by preparative SFC using the following conditions (column: CHIRAL ART Cellulose-SB, 3*25 cm, 5 μm; mobile phase A: CO2, mobile phase B: IPA:DCM=1:1--HPLC; flow rate: 90 mL / min; gradient: isocratic 60% B; column temperature (°C): 35; back pressure (bar): 100; wavelength: 277 nm; RT1 (min): 3.33; RT2 (min): 4.28; sample solvent: MeOH:DCM=1:1--HPLC; injection volume: 1 mL) to provide the title compound (second peak) (36.9 mg, 35.46%) as a white solid. 1HNMR (400MHz, DMSO-d6) δ12.30–12.25(m,1H),9.01–8.96(m,1H),8.89–8.79(m,2H),8.75(s,1H),8.44(d,J=7.7Hz,1H),8.01(d,J=7.9,1.7 Hz,1H),7.50(s,1H),7.47–7.42(m,2H),7.40–7.34(m,3H),7.07–6.92(m,3H),5.11(d,J=3.6Hz,1H),4.94(q,J=7.2,6.5Hz,1H),4.39(t,J=7 .9Hz,1H),4.30(s,1H),4.24–4.07(m,2H),3.93–3.59(m,5H),3.51(d, J=10.7Hz,1H),2.65–2.56(m,1H),2.48–2.42(m,3H),2.38–2.23(m,2H) ),2.10–1.99(m,1H),1.85–1.69(m,1H),1.44(dd,J=40.6,7.0Hz,3H),1.05–0.98(m,3H),0.89–0.81(m,3H).LCMS(ESI)m / z:[M+H]+=856.40.

[1405] The compounds in Table 5 were prepared using a procedure analogous to that used above to prepare (2S,4R)-4-hydroxy-1-[(2R)-2-(3-{2-[(3R)-3-[3-(2-hydroxyphenyl)thieno[2,3-c]pyridazin-6-yl]pyrrolidin-1-yl]pyrimidin-5-yl}-1,2-oxazol-5-yl)-3-methylbutanoyl]-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (Compound 5) using the appropriate aryl bromide and amine.

[1406] Table 5.

[1407]

[1408]

[1409]

[1410]

[1411]

[1412]

[1413]

[1414]

[1415]

[1416]

[1417]

[1418]

[1419]

[1420]

[1421]

[1422]

[1423]

[1424]

[1425]

[1426]

[1427] Preparation of (2S,4R)-4-hydroxy-1-{2-[3-(2-{4-[3-(2-hydroxyphenyl)thieno[3,2-c]pyridazin-6-yl]piperazin-1-yl}pyrimidin-5-yl)-1,2-oxazol-5-yl]-3-methylbutyryl}-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (Compound 9)

[1428]

[1429] Step 1: Preparation of 4-bromo-6-chloro-3-iodopyridazine (Intermediate 2)

[1430]

[1431] To a stirred solution of 4-bromo-6-chloropyridazine-3-amine (20.00 g, 95.951 mmol, 1.00 equivalent) in THF (200.00 mL) was added CuI (21.93 g, 115.141 mmol, 1.20 equivalent) and CH2I2 (30.84 g, 115.141 mmol, 1.20 equivalent) at room temperature. T-BuNO2 (11.87 g, 115.141 mmol, 1.20 equivalent) was added to the above mixture under a nitrogen atmosphere at room temperature. The resulting mixture was stirred for another 6 hours at 60 ° C under a nitrogen atmosphere. The mixture was cooled to room temperature. The reaction was quenched with water at 0 ° C. The resulting mixture was extracted with EA (2x200 mL). The combined organic layer was washed with saturated brine (1x200 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (1 / 1) to provide Intermediate 2 (15 g, 48.96%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=319.

[1432] Step 2: Preparation of 4-(tert-butylsulfanyl)-6-chloro-3-iodopyridazine (Intermediate 4)

[1433]

[1434] To a stirred solution of intermediate 2 (15.00 g, 46.975 mmol, 1.00 equiv) in THF (100.00 mL) was added 2-methyl-2-propanethiol (5.08 g, 56.370 mmol, 1.20 equiv) and NaH (2.25 g, 93.950 mmol, 2.00 equiv, 60%) at 0 ° C. The resulting mixture was stirred at 60 ° C for 1 hour. The reaction was quenched with water at 0 ° C. The resulting mixture was extracted with EA (2x 200 mL). The combined organic layer was washed with saturated brine (1x200 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with PE / EA (1 / 1) to provide intermediate 4 (6.4 g, 41.46%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=329.

[1435] Step 3: Preparation of tert-butyl 2-[4-(tert-butylsulfanyl)-6-chloropyridazin-3-yl]-2-cyanoacetate (Intermediate 6)

[1436]

[1437] To a stirred solution of intermediate 4 (6.40 g, 19.477 mmol, 1.00 equiv) and tert-butyl 2-cyanoacetate (5.50 g, 38.954 mmol, 2.00 equiv) in 1,4-dioxane (60.00 mL) was added CuI (0.74 g, 3.895 mmol, 0.20 equiv), picolinic acid (1.20 g, 9.739 mmol, 0.50 equiv) and Cs2CO3 (19.04 g, 58.431 mmol, 3.00 equiv) at room temperature. The resulting mixture was stirred at 100 °C under nitrogen atmosphere for 2 hours. The mixture was cooled to room temperature. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, H2O (0.1% FA) / ACN, gradient from 0 to 100% in 30 minutes; detector, UV 254 / 220 nm. This produced intermediate 6 (5 g, 75.10%) as a brown oil. LCMS (ESI) m / z: [M+H]+=342.

[1438] Step 4: Preparation of 2-[4-(tert-butylsulfanyl)-6-chloropyridazin-3-yl]acetonitrile (Intermediate 7)

[1439]

[1440] To HFIP (50.00 mL) was added intermediate 6 (5.00 g, 14.626 mmol, 1.00 equiv) at room temperature. The resulting mixture was stirred at 60 °C for 16 hours. The residue was allowed to cool to room temperature. The resulting mixture was concentrated under vacuum. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, H2O (0.1% FA) / ACN, 0 to 100% gradient in 30 minutes; detector, UV254 / 220 nm. This produced intermediate 7 (1.2 g, 33.94%) as a brown oil. LCMS (ESI) m / z: [M+H]+=242.

[1441] Step 5: Preparation of 3-chlorothieno[3,2-c]pyridazin-6-amine (Intermediate 8)

[1442]

[1443] Intermediate 7 (1.20 g, 4.964 mmol, 1.00 equiv) was added to trifluoroacetaldehyde (10.00 mL) at room temperature. The resulting mixture was stirred at 100°C for 16 hours. The mixture was allowed to cool to room temperature. The resulting mixture was concentrated under vacuum. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, H2O (10 mmol / L NH4HCO3) / ACN, 0 to 100% gradient in 30 minutes; detector, UV 254 / 220 nm. This produced intermediate 8 (283 mg, 30.71%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=186.

[1444] Step 6: Preparation of 3,6-dichlorothieno[3,2-c]pyridazine (Intermediate 9)

[1445]

[1446] To a stirred solution of intermediate 8 (283.00 mg, 1.525 mmol, 1.00 equiv) in ACN (10.00 mL) was added CuCl (301.86 mg, 3.050 mmol, 2.00 equiv) at 0°C. To the above mixture was added t-BuNO2 (314.42 mg, 3.050 mmol, 2.00 equiv) at 0°C. The resulting mixture was stirred for another 1 hour at room temperature. The reaction was quenched with water at 0°C. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, H2O (0.1% FA) / ACN, 0 to 100% gradient in 30 minutes; detector, UV 254 / 220 nm. This produced intermediate 9 (153 mg, 48.94%) as a brown solid. LCMS (ESI) m / z: [M+H]+=205.

[1447] Step 7: Preparation of 1-{3-chlorothieno[3,2-c]pyridazin-6-yl}piperazine (Intermediate 11)

[1448]

[1449] To a stirred solution of intermediate 9 (153.00 mg, 0.746 mmol, 1.00 equiv) in DMSO (5.00 mL) was added piperazine (192.81 mg, 2.238 mmol, 3.00 equiv) and DIEA (482.17 mg, 3.730 mmol, 5.00 equiv) at room temperature. The resulting mixture was stirred at 100 °C for 1 hour. The mixture was allowed to cool to room temperature. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, H2O (0.1% FA) / ACN, 0 to 100% gradient in 30 minutes; detector, UV 254 / 220 nm. This produced intermediate 11 (114 mg, 59.98%) as a brown solid. LCMS (ESI) m / z: [M+H]+=255.

[1450] Step 8: Preparation of 2-[6-(piperazin-1-yl)thieno[3,2-c]pyridazin-3-yl]phenol (Intermediate 13)

[1451]

[1452] To a stirred solution of intermediate 11 (114.00 mg, 0.448 mmol, 1.00 equiv) and 2-hydroxyphenylboronic acid (185.18 mg, 1.344 mmol, 3.00 equiv) in 1,4-dioxane (8.00 mL) and H2O (2.00 mL) was added XPhos Pd G3 (37.88 mg, 0.045 mmol, 0.10 equiv) and Cs2CO3 (437.43 mg, 1.344 mmol, 3.00 equiv) at room temperature. The resulting mixture was stirred at 80°C under nitrogen atmosphere for 6 hours. The mixture was cooled to room temperature. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, H2O (0.1% FA) / ACN, gradient from 0 to 100% in 30 minutes; detector, UV 254 / 220 nm. This produced intermediate 13 (44 mg, 31.47%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=313.

[1453] Step 9: Preparation of methyl 2-[3-(2-{4-[3-(2-hydroxyphenyl)thieno[3,2-c]pyridazin-6-yl]piperazin-1-yl}pyrimidin-5-yl)-1,2-oxazol-5-yl]-3-methylbutanoate (Intermediate 15)

[1454]

[1455] To a stirred solution of intermediate 13 (44.00 mg, 0.141 mmol, 1.00 equiv) in DMSO (5.00 mL) was added methyl 2-[3-(2-chloropyrimidin-5-yl)-1,2-oxazol-5-yl]-3-methylbutanoate (41.74 mg, 0.141 mmol, 1.00 equiv) and DIEA (54.57 mg, 0.423 mmol, 3.00 equiv) at room temperature. The resulting mixture was stirred at 100 °C for 1 hour. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, H2O (0.1% FA) / ACN, gradient from 0 to 100% in 30 minutes; detector, UV 254 / 220 nm. This produced intermediate 15 (19 mg, 23.75%) as a brown solid. LCMS (ESI) m / z: [M+H]+=572.

[1456] Step 10: Preparation of 2-[3-(2-{4-[3-(2-hydroxyphenyl)thieno[3,2-c]pyridazin-6-yl]piperazin-1-yl}pyrimidin-5-yl)-1,2-oxazol-5-yl]-3-methylbutanoic acid (Intermediate 16)

[1457]

[1458] To a stirred solution of intermediate 15 (19.00 mg, 0.033 mmol, 1.00 equiv) in MeOH (2.00 mL) and H2O (2.00 mL) was added LiOH·H2O (14.52 mg, 0.330 mmol, 10.00 equiv) at room temperature. The resulting mixture was stirred at room temperature for 16 hours. The mixture was acidified to pH 3 with HCl (1 M). The resulting mixture was concentrated under vacuum. This produced intermediate 16 (20 mg, crude) as a yellow solid. LCMS (ESI) m / z: [M+H]+=558.

[1459] Step 11: Preparation of (2S,4R)-4-hydroxy-1-{2-[3-(2-{4-[3-(2-hydroxyphenyl)thieno[3,2-c]pyridazin-6-yl]piperazin-1-yl}pyrimidin-5-yl)-1,2-oxazol-5-yl]-3-methylbutyryl}-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide

[1460]

[1461] To a stirred solution of intermediate 16 (20.00 mg, 0.036 mmol, 1.00 equiv) in DMF (1.00 mL) was added PyBOP (55.99 mg, 0.108 mmol, 3.00 equiv) and DIEA (23.18 mg, 0.180 mmol, 5.00 equiv) at room temperature. To the above mixture was added (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (11.89 mg, 0.036 mmol, 1.00 equiv) at room temperature. The resulting mixture was stirred at room temperature for another 16 hours. The residue was purified by preparative HPLC under the following conditions: column: XBridge Prep OBD C18 column, 30*150 mm, 5 μm; mobile phase A: water (10 mmol / L NH4HCO3), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 42% B to 65% B, 65% B in 9 minutes; wavelength: 254 / 220 nm; RT1 (min): 8.47; number of runs: 0. This produced the title compound (0.7 mg, 2.16%) as a yellow solid. 1HNMR(400MHz,DMSO-d6)δ13.81–13.76(m,1H),9.01–8.87(m,3H),8.84(s,1H),8.45–8.23(m,1H),7.98–7.93(m,1H),7.51–7.4 2(m,1H),7.40–7.26(m,4H),7.01–6.94(m,3H),6.71(s,1H),6.15–4.98(m,1H),4.99–4.84(m,1H),4.60–4.36(m,1H),4.34–4.2 2(m,1H),4.10–3.99(m,4H),3.99–3.82(m,1H),3.68–3.57(m,5H),3.40–3.34(m,1H),2.48–2.43(m,2H),2.40(s,2H),2.11–1.9 9(m,1H),1.85–1.73(m,1H),1.34(d,J=7.0Hz,3H),1.01(d,J=6.6Hz,3H),0.86(d,J=6.6Hz,3H).LCMS(ESI)m / z:[M+H]+=871.20.

[1462] Preparation of (2S,4R)-4-hydroxy-1-[(2R)-2-[3-(4-{5-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]pyrimidin-2-yl}piperazin-1-yl)-1,2-oxazol-5-yl]-3-methylbutyryl]-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (Compound 31)

[1463]

[1464] Step 1: Preparation of ethyl 2-cyano-2-(3,6-dichloropyridazin-4-yl)-2-methylacetate (Intermediate 2)

[1465]

[1466] To a solution of 3,4,6-trichloropyridazine (10 g, 54.520 mmol, 1 eq.) and ethyl 2-cyanopropionate (6.93 g, 54.520 mmol, 1 eq.) in DMSO (80 mL) was added DIEA (21.14 g, 163.560 mmol, 3 eq.) and the resulting solution was stirred at 100 °C for 3 hours. The mixture was diluted with EtOAc (200 mL) and washed with water (200 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by flash C18 chromatography with an elution gradient of 0 to 35% ACN / H2O to give intermediate 2 (7.3 g, 48.85%) as a brown oil. LCMS (ESI) m / z: [M+H]+=274.

[1467] Step 2: Preparation of 2-(3,6-dichloropyridazin-4-yl)propionitrile (Intermediate 3)

[1468]

[1469] To a solution of intermediate 2 (7.3 g, 26.633 mmol, 1 eq.) and NaCl (3.11 g, 53.266 mmol, 2 eq.) in DMSO (50 mL) and H2O (10 mL), the resulting solution was stirred at 120 °C for 16 hours. The mixture was diluted with EtOAc (100 mL) and washed with water (100 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by flash C18 chromatography with an elution gradient of 0 to 42% ACN / H2O to give intermediate 3 (2.8 g, 52.04%) as a brown oil. LCMS (ESI) m / z: [M+H]+=202.

[1470] Step 3: Preparation of 3-chloro-5-methylthieno[2,3-c]pyridazin-6-amine (Intermediate 4)

[1471]

[1472] To a solution of intermediate 3 (2.8 g, 12.374 mmol, 1 eq.) and CuI (235.66 mg, 1.237 mmol, 0.1 eq.) in DMF (25 mL) was added TMEDA (287.59 mg, 2.475 mmol, 0.20 eq.) and Na2S.9H2O (8.92 g, 37.122 mmol, 3 eq.) and the resulting solution was stirred at 80 °C for 2 hours (under N2 atmosphere). The resulting mixture was filtered and the filter cake was washed with EtOAc (3x20 mL). The filtrate was concentrated under reduced pressure. The crude product was purified by flash C18 chromatography with an elution gradient of 0 to 42% MeOH / H2O to afford intermediate 4 (1.1 g, 44.52%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=200.

[1473] Step 4: Preparation of 6-bromo-3-chloro-5-methylthieno[2,3-c]pyridazine (Intermediate 5)

[1474]

[1475] To a solution of intermediate 4 (1.1 g, 5.509 mmol, 1 eq.) and CuBr2 (1.48 g, 6.611 mmol, 1.2 eq.) in MeCN (10 mL) was added 2-methyl-2-propionitrile (681.76 mg, 6.611 mmol, 1.2 eq.) and the resulting solution was stirred at 25 °C for 2 hours. The mixture was diluted with EtOAc (200 mL) and washed with water (200 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by flash C18 chromatography with an elution gradient of 0 to 64% ACN / H2O to give intermediate 5 (693 mg, 47.73%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=263.

[1476] Step 5: Preparation of 5-{3-chloro-5-methylthieno[2,3-c]pyridazin-6-yl}-2-methoxypyrimidine (Intermediate 6)

[1477]

[1478] To a solution of intermediate 5 (693 mg, 2.630 mmol, 1 eq) and 2-methoxypyrimidin-5-ylboronic acid (404.77 mg, 2.630 mmol, 1 eq) in dioxane (6 mL) and H2O (1.5 mL) were added palladium dichloride; bis(triphenylphosphine) (184.57 mg, 0.263 mmol, 0.1 eq) and K3PO4 (1.12 g, 5.260 mmol, 2 eq) and the resulting solution was stirred at 80 °C for 16 hours (under N2 atmosphere). The mixture was diluted with EtOAc (100 mL) and washed with water (100 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product. The crude product was purified by flash C18 chromatography with an elution gradient of 0 to 67% ACN / H2O to afford intermediate 6 (253 mg, 32.87%) as a brown solid. LCMS (ESI) m / z: [M+H]+=293.

[1479] Step 6: Preparation of 2-[6-(2-methoxypyrimidin-5-yl)-5-methylthieno[2,3-c]pyridazin-3-yl]phenol (Intermediate 7)

[1480]

[1481] To a solution of intermediate 6 (271 mg, 0.926 mmol, 1 eq) and 2-hydroxyphenylboronic acid (191.53 mg, 1.389 mmol, 1.5 eq) in dioxane (6 mL) and H2O (1.5 mL) was added XPhos Pd G3 (78.36 mg, 0.093 mmol, 0.1 eq) and Cs2CO3 (603.24 mg, 1.852 mmol, 2 eq) and the resulting solution was stirred at 80 °C for 2 hours (under N2 atmosphere). The mixture was diluted with EtOAc (80 mL) and washed with water (80 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by flash silica gel column chromatography with an elution gradient of 0 to 51% EA / PE to give intermediate 7 (234 mg, 72.14%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=351.

[1482] Step 7: Preparation of 2-[6-(2-chloropyrimidin-5-yl)-5-methylthieno[2,3-c]pyridazin-3-yl]phenol (Intermediate 8)

[1483]

[1484] To a solution of intermediate 7 (323 mg, 0.922 mmol, 1 eq.) in POCl3 (0.5 mL) was added DMF (3 mL) dropwise at 0°C and the resulting solution was stirred at 60°C for 2 hours. The reaction was quenched with water at 0°C. The mixture was diluted with EtOAc (100 mL) and washed with water (100 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by flash C18 chromatography with an elution gradient of 0 to 62% ACN / H2O to give intermediate 8 (108 mg, 33.02%) as a yellow solid. LCMS (ESI) m / z: [M+H]+=355.

[1485] Step 8: Preparation of methyl 2-[3-(4-{5-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]pyrimidin-2-yl}piperazin-1-yl)-1,2-oxazol-5-yl]-3-methylbutanoate (Intermediate 9)

[1486]

[1487] To a solution of intermediate 8 (103 mg, 0.290 mmol, 1 eq) and methyl 3-methyl-2-[3-(piperazin-1-yl)-1,2-oxazol-5-yl]butanoate (93.13 mg, 0.348 mmol, 1.2 eq) in DMSO (3 mL) was added DIEA (112.56 mg, 0.870 mmol, 3 eq) and the resulting solution was stirred at 100 °C for 4 hours. The reaction was purified by flash C18 chromatography with an elution gradient of 0 to 71% ACN / H2O to afford intermediate 9 (96 mg, 56.46%) as a yellow solid. LCMS (ESI) m / z: [M+H]+ = 586.

[1488] Step 9: Preparation of 2-[3-(4-{5-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]pyrimidin-2-yl}piperazin-1-yl)-1,2-oxazol-5-yl]-3-methylbutanoic acid (Intermediate 10)

[1489]

[1490] To a solution of methyl intermediate 9 (91 mg, 0.155 mmol, 1 eq.) and LiOH (7.44 mg, 0.310 mmol, 2 eq.) in MeOH (4 mL) and H2O (1 mL). The resulting solution was stirred at 25 °C for 16 hours. The mixture was diluted with EtOAc (50 mL) and washed with water (50 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give intermediate 10 (92 mg, crude), which was used directly in the next step without further purification. LCMS (ESI) m / z: [M+H]+=572.#

[1491] Step 10: Preparation of (2S,4R)-4-hydroxy-1-{2-[3-(4-{5-[3-(2-hydroxyphenyl)-5-methylthieno[2,3-c]pyridazin-6-yl]pyrimidin-2-yl}piperazin-1-yl)-1,2-oxazol-5-yl]-3-methylbutanoyl}-N-[(1S)-1-[4-(4-methyl-1,3-thiazo...

Claims

1. A compound having a structure of Formula I or II or a pharmaceutically acceptable salt thereof: in Ring system A is a 5- to 9-membered heterocyclyl or heteroaryl; m is 0, 1, 2, or 3; k is 0, 1, or 2; Each R 1 is independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C3-C8 cycloalkyl or optionally substituted CH2-C3-C8 cycloalkyl; each X is independently halogen; L is a linker; and B is the degradation part.

2. The compound of claim 1 or a pharmaceutically acceptable salt thereof, which has a structure of Formula I or II: in Ring system A is a 5- to 9-membered heterocyclyl or heteroaryl; m is 0, 1, 2, or 3; k is 0, 1, or 2; L is the connector; B is the degradation part; Each R 1 is independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8 cycloalkyl or optionally substituted C2-C9 heterocyclyl; and Each X is independently halogen.

3. The compound of claim 1 or a pharmaceutically acceptable salt thereof, which has a structure of Formula I or II: in Ring system A is a 5- to 9-membered heterocyclyl or heteroaryl; m is 0, 1, 2, or 3; k is 0, 1, or 2; L is a linker of formula III: A 1 -(B 1 ) f -(C 1 ) g -(B 2 ) h -(B 3 ) i -(C 2 ) j -(B 4 ) k –A 2 , Formula III in A 1 is the bond between the linker and the ring system A; A 2 is the bond between the degradable moiety and the linker; B 1 , B 2 , B 3 and B 4 are each independently an optionally substituted ethynyl, an optionally substituted C6-C 10 Aryl, optionally substituted C3-C 10 cycloalkyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C2-C9 heteroaryl, O, S, S(O)2 or NR N ; Each R N are independently H, optionally substituted C 1–4 Alkyl, optionally substituted C 2–4 alkenyl, optionally substituted C2-4 alkynyl, optionally substituted C 2–6 Heterocyclic group, optionally substituted C 6–12 Aryl or optionally substituted C 1–7 heteroalkyl; C 1 and C 2 are each independently carbonyl, thiocarbonyl, sulfonyl or phosphoryl; and f, g, h, i, j and k are each independently 0 or 1. B is the degradation part; Each R 1 is independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8 cycloalkyl or optionally substituted C2-C9 heterocyclyl; and Each X is independently halogen.

4. The compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, wherein the compound has the structure of Formula IA or II-A: The dashed bonds represent single bonds or double bonds.

5. The compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, wherein the compound has a structure of formula IG or II-G:

6. The compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, wherein the compound has a structure of Formula IH or II-H:

7. The compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, wherein m is 0 or 1.

8. The compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, wherein m is 1.

9. The compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, wherein R 1 is halogen, optionally substituted C1-C6 alkyl or optionally substituted C3-C8 cycloalkyl.

10. The compound according to claim 9 or a pharmaceutically acceptable salt thereof, wherein R 1 It's methyl.

11. The compound according to claim 9 or a pharmaceutically acceptable salt thereof, wherein R 1 It's cyclopropane.

12. The compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, wherein m is 0.

13. The compound according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, wherein k is 1.

14. The compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, wherein X is Cl.

15. The compound according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, wherein k is 0.

16. The compound of any one of claims 1 to 15, or a pharmaceutically acceptable salt thereof, wherein L is a linker of formula III: A 1 -(B 1 ) f -(C 1 ) g -(B 2 ) h -(B 3 ) i -(C 2 ) j -(B 4 ) k –A 2 , Formula III in A 1 is the bond between the linker and the ring system A; A 2 is the bond between the degradable moiety and the linker; B 1 , B 2 , B 3 and B 4 are each independently an optionally substituted ethynyl, an optionally substituted C6-C 10 aryl, optionally substituted C3-C10 cycloalkyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C2-C9 heteroaryl, O, S, S(O)2 or NR N ; Each R N are independently H, optionally substituted C 1–4 Alkyl, optionally substituted C 2–4 Alkenyl, optionally substituted C 2–4 Alkynyl, optionally substituted C 2–6 Heterocyclic group, optionally substituted C 6–12 Aryl or optionally substituted C 1–7 heteroalkyl; C 1 and C 2 are each independently carbonyl, thiocarbonyl, sulfonyl or phosphoryl; and f, g, h, i, j and k are each independently 0 or 1.

17. The compound of any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, wherein the linker has the following structure: A 1 -(B 1 ) f -(B 2 ) h -(B 3 ) i -(B 4 ) k –A 2 , Among them B 1 , B 2 , B 3 and B 4 are each independently an optionally substituted ethynyl, an optionally substituted C6-C 10 Aryl, optionally substituted C3-C 10 cycloalkyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C2-C9 heteroaryl, O or NR N .

18. The compound according to any one of claims 16 to 17, or a pharmaceutically acceptable salt thereof, wherein at least one of f, h, i and k is 1.

19. The compound according to any one of claims 16 to 18, or a pharmaceutically acceptable salt thereof, wherein B 1 , B 2 , B 3 and B 4 are each independently O, ethynyl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C9 heterocyclyl, optionally substituted C3-C 10 Cycloalkyl or optionally substituted C6-C 10 Aryl.

20. The compound according to any one of claims 16 to 19, or a pharmaceutically acceptable salt thereof, wherein B 1 and B 4 Each independently is 21. The compound of claim 20 or a pharmaceutically acceptable salt thereof, wherein B 1 yes 22. The compound according to claim 20 or 21, or a pharmaceutically acceptable salt thereof, wherein B 4 yes 23. The compound according to any one of claims 16 to 17, or a pharmaceutically acceptable salt thereof, wherein B 2 It is NH, 24. The compound of any one of claims 16 to 23, or a pharmaceutically acceptable salt thereof, wherein g, h, I and j are 0.

25. The compound according to any one of claims 1 to 24 or a pharmaceutically acceptable salt thereof, wherein the degradation portion B has a structure of formula A-1: in Y 1 yes R A5 is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; R A6 is H or optionally substituted C1-C6 alkyl; and R A7 is H or optionally substituted C1-C6 alkyl; or R A6 and R A7 and R are combined together with the carbon atoms to which they are each bonded to form an optionally substituted C3-C6 carbocyclyl or an optionally substituted C2-C5 heterocyclyl; or A6 and R A7 Combined together with the carbon atom to which each is bonded to form an optionally substituted C3-C6 carbocyclyl or an optionally substituted C2-C5 heterocyclyl; R A8 is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; R A1 , R A2 , R A3 and R A4 Each independently is H, A 2 , halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclic group, optionally substituted C2-C9 heterocyclic group, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, optionally substituted -O-C3-C6 carbocyclyl, hydroxyl, thiol, or optionally substituted amino; or R A1 and R A2 , R A2 and R A3 , and / or R A3 and R A4 Combined with the carbon atoms to which they are attached to form and is an optionally substituted C6-C10 aryl group, an optionally substituted C3-C 10 carbocyclic group, optionally substituted C2-C9 heteroaryl group or C2-C9 heterocyclic group, any of which is optionally substituted by A 2 replace, Where R A1 , R A2 , R A3 and R A4 One of them is A 2 ,or A 2 Replacement; and A 2 is the bond between the degradable moiety and the linker.

26. The compound of claim 25 or a pharmaceutically acceptable salt thereof, wherein the degradation portion has the following structure:

27. The compound of any one of claims 1 to 24, or a pharmaceutically acceptable salt thereof, wherein the degradation moiety has a structure of Formula C: in L 4 Yes-N(R B1 )(R B2 ), R B1 It is H, A 2 , optionally substituted C1-C6 alkyl or optionally substituted C1-C6 heteroalkyl; R B2 is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; R B3 Yes A 2 , optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; R B4 is H, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; R B5 is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; v2 is 0, 1, 2, 3, or 4; Each R B6 Independently is A 2 , halogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkynyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclic group, optionally substituted C2-C9 heterocyclic group, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxy, thiol, cyano or optionally substituted amino; R B7 and R B8 are each independently H, halogen, optionally substituted C1-C6 alkyl or optionally substituted C6-C 10 Aryl; R B9 is H or optionally substituted C1-C6 alkyl; R B10 is H or F; and A 2 is the bond between the degradable moiety and the linker; Where R B1 , R B3 and R B6 One and only one of them is A2, or a pharmaceutically acceptable salt thereof.

28. The compound of any one of claims 1 to 24, or a pharmaceutically acceptable salt thereof, wherein the degradation moiety has a structure of Formula C: in L 4 Yes-N(R B1 )(R B2 ), R B1 It is H, A 2 , optionally substituted C1-C6 alkyl or optionally substituted C1-C6 heteroalkyl; R B2 is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; R B3 Yes A 2 , optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; R B4 is H, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; R B5 is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; v2 is 0, 1, 2, 3, or 4; Each R B6 Independently is A 2 , halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclic group, optionally substituted C2-C9 heterocyclic group, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxy, thiol or optionally substituted amino; R B7 and R B8 are each independently H, halogen, optionally substituted C1-C6 alkyl or optionally substituted C6-C 10 Aryl; R B9 is H or optionally substituted C1-C6 alkyl; and A 2 is the bond between the degradable moiety and the linker; Where R B1 , R B3 and R B6 One and only one of them is A2, or a pharmaceutically acceptable salt thereof.

29. The compound of any one of claims 27-28 or a pharmaceutically acceptable salt thereof, wherein the degradation portion is 30. The compound of claim 27 or a pharmaceutically acceptable salt thereof, wherein the degraded portion is 31. The compound of any one of claims 27-28 or a pharmaceutically acceptable salt thereof, wherein the degradation portion is 32. The compound of any one of claims 27 or a pharmaceutically acceptable salt thereof, wherein the degradation portion is 33. The compound of claim 27 or a pharmaceutically acceptable salt thereof, wherein the degraded portion is 34. The compound of claim 27 or a pharmaceutically acceptable salt thereof, wherein the degraded portion is 35. The compound of any one of claims 1-24 or a pharmaceutically acceptable salt thereof, wherein the degradation portion has a structure of formula C5: in L4 is -N(R B1 )(R B2 ), R B1 It is H, A 2 , optionally substituted C1-C6 alkyl or optionally substituted C1-C6 heteroalkyl; R B2 is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; R B3 Yes A 2 , optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclic group, optionally substituted C6-C10 aryl group, optionally substituted C1-C6 alkyl group, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; R B5 is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; v2 is 0, 1, 2, 3, or 4; Each R B6 Independently is A 2 , halogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkynyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclic group, optionally substituted C2-C9 heterocyclic group, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxy, thiol, cyano or optionally substituted amino; R B7 and R B8 Each is independently H, halogen, optionally substituted C1-C6 alkyl or optionally substituted C6-C10 aryl; R B9 is H or optionally substituted C1-C6 alkyl; R B11 is H, alcohol, organic boronic acid, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; and A 2 is the bond between the degradable moiety and the linker; Where R B1 , R B3 and R B6 One and only one of them is A 2 , or a pharmaceutically acceptable salt thereof.

36. The compound of claim 35 or a pharmaceutically acceptable salt thereof, wherein the degraded portion is 37. The compound of any one of claims 1 to 24, or a pharmaceutically acceptable salt thereof, wherein the degradation moiety has a structure of Formula D: in L4 is -N(R B1 )(R B2 ), R B1 It is H, A 2 , optionally substituted C1-C6 alkyl or optionally substituted C1-C6 heteroalkyl; R B2 is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; R B3 Yes A 2 , optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10芳 base; R B4 is H, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; R B5 is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; v2 is 0, 1, 2, 3, or 4; Each R B6 Independently is A 2 , halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 alkynyl, optionally substituted C3-C 10 carbocyclic group, optionally substituted C2-C9 heterocyclic group, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxy, thiol, cyano or optionally substituted amino; R B9 is H or optionally substituted C1-C6 alkyl; and A 2 is the bond between the degradable moiety and the linker; Where R B1 , R B3 and R B6 One and only one of them is A 2 , or a pharmaceutically acceptable salt thereof.

38. The compound of claim 37 or a pharmaceutically acceptable salt thereof, wherein the degraded portion is 39. The compound of claim 37 or a pharmaceutically acceptable salt thereof, wherein the degraded portion is 40. The compound of any one of claims 1-24, or a pharmaceutically acceptable salt thereof, wherein the degradation moiety has a structure of formula Da: in L 4 Yes-N(R B1 )(R B2 ), R B1 It is H, A 2 , optionally substituted C1-C6 alkyl or optionally substituted C1-C6 heteroalkyl; R B2 is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; R B3 Yes A 2 , optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; R B4 is H, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; R B5 is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; X 1 and X 2 are each independently C, N or O; v2 is 0, 1, 2, 3, or 4; Each R B6 Each independently is A 2 , halogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkynyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclic group, optionally substituted C2-C9 heterocyclic group, optionally substituted C 6- C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxy, thiol, cyano or optionally substituted amino; R B9 is H or optionally substituted C1-C6 alkyl; and A 2 is the bond between the degradable moiety and the linker; Where R B1 , R B3 and R B6 One and only one of them is A2, or a pharmaceutically acceptable salt thereof.

41. The compound of claim 40 or a pharmaceutically acceptable salt thereof, wherein the degraded portion is 42. The compound of any one of claims 1-24 or a pharmaceutically acceptable salt thereof, wherein the degradation moiety has a structure of Formula E: in L 4 Yes-N(R B1 )(R B2 ), R B1 It is H, A 2 , optionally substituted C1-C6 alkyl or optionally substituted C1-C6 heteroalkyl; R B2 is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; R B3 Yes A 2 , optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; R B4 is H, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; R B5 is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; R B9 is H, optionally substituted C1-C6 alkyl, optionally substituted C3-C6 alkynyl, optionally substituted C3-C 10 Carbocyclic or optionally substituted C2-C 10 Heterocyclic group; B 10 is H, optionally substituted C1-C6 alkyl, optionally substituted C3-C6 alkynyl, optionally substituted C3-C 10 Carbocyclic, optionally substituted C2-C 10 heterocyclic group, optionally substituted amino group or cyano group, and A 2 is the bond between the degradable moiety and the linker; Where R B1 , R B3 and R B6 One and only one of them is A 2 , or a pharmaceutically acceptable salt thereof.

43. The compound of claim 42 or a pharmaceutically acceptable salt thereof, wherein the degraded portion is 44. The compound of any one of claims 1-24, or a pharmaceutically acceptable salt thereof, wherein the degradation moiety has a structure of Formula F: in L 4 Yes-N(R B1 )(R B2 ), R B1 It is H, A 2 , optionally substituted C1-C6 alkyl or optionally substituted C1-C6 heteroalkyl; R B2 is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; R B3 Yes A 2 , optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; R B4 is H, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; R B5 is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; A 2 is the bond between the degradable moiety and the linker; Where R B1 or R B3 One and only one of them is A 2 , or a pharmaceutically acceptable salt thereof.

45. The compound of claim 44 or a pharmaceutically acceptable salt thereof, wherein the degraded portion is 46. ​​The compound of claim 44 or a pharmaceutically acceptable salt thereof, wherein the degraded portion is 47. The compound of any one of claims 1 to 15 and 25 to 46, or a pharmaceutically acceptable salt thereof, wherein the linker has the structure of Formula II: A 1 -(B 1 ) f -(C 1 ) g -(B 2 ) h -(D)-(B 3 ) i -(C 2 ) j -(B 4 ) k –A 2 , Formula II or a pharmaceutically acceptable salt thereof, in A 1 is the bond between the linker and the ring system A; A 2 is the bond between the degradable moiety and the linker; B 1 , B 2 , B 3 and B 4 are each independently an optionally substituted C1-C4 alkyl, an optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 Aryl C 1-4 alkyl, optionally substituted C1-C4 heteroalkyl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C3-C 10 Carbocyclic, optionally substituted C2-C 10 Heterocyclyl, optionally substituted C2-C6 heteroaryl, optionally substituted C 6–12 Aryl, O, S, S(O)2 or NR N ; Each R N are independently H, optionally substituted C 1–4 Alkyl, optionally substituted C 2–4 Alkenyl, optionally substituted C 2–4 Alkynyl, optionally substituted C 2–10 Heterocyclic group, optionally substituted C 2–6 Heteroaryl or optionally substituted C 1–7 heteroalkyl; C1 and C2 are each independently carbonyl, thiocarbonyl, sulfonyl or phosphoryl; f, g, h, i, j and k are each independently 0 or 1; and D is optionally substituted C 1–10 Alkyl, optionally substituted C 2–10 Alkenyl, optionally substituted C 2–10 Alkynyl, optionally substituted C 2–10 Heterocyclic group, optionally substituted C 2–6 Heteroaryl, optionally substituted C 6–12 Aryl, optionally substituted C2-C 10 Polyethylene glycol, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C3-C 10 Carbocyclic or optionally substituted C 1–10 heteroalkyl; or D is absent, and the linker is A 1 -(B 1 )f-(C 1 )g-(B 2 )h-(B 3 )i-(C 2 )j-(B 4 )k–A 2 .

48. The compound of claim 47 or a pharmaceutically acceptable salt thereof, wherein A1 is the bond between the linker and the benzopyridazine core ring system; A2 is the bond between the degradable moiety and the linker; B 1 , B 2 , B 3 and B 4 are each independently an optionally substituted C1-C4 alkyl, an optionally substituted C6-C 10 Aryl, optionally substituted C6-C 10 Aryl C 1-4 alkyl, optionally substituted C1-C4 heteroalkyl, optionally substituted C3-C 10 cycloalkyl, optionally substituted C2-C8 heterocyclyl, optionally substituted C2-C6 heteroaryl, optionally substituted C 6–12 Aryl, O, S, S(O)2 or NR N ; Each R N are independently H, optionally substituted C 1–4 Alkyl, optionally substituted C 2–4 Alkenyl, optionally substituted C 2–4 Alkynyl, optionally substituted C 2–6 Heterocyclic group, optionally substituted C 2–6 Heteroaryl or optionally substituted C 1–7 heteroalkyl; C 1 and C 2 are each independently carbonyl, thiocarbonyl, sulfonyl or phosphoryl; f, g, h, i, j and k are each independently 0 or 1; and D is optionally substituted C 1–10 Alkyl, optionally substituted C 2–10 Alkenyl, optionally substituted C 2–10 Alkynyl, optionally substituted C 2–6 Heterocyclic group, optionally substituted C 2–6 Heteroaryl, optionally substituted C 6–12 Aryl, optionally substituted C2-C 10 Polyethylene glycol or optionally substituted C 1–10 heteroalkyl; or D is absent, and the linker is A 1 -(B 1 )f-(C 1 )g-(B 2 )h-(B 3 )i-(C 2 )j-(B 4 )k–A 2 .

49. The compound of claim 47, or a pharmaceutically acceptable salt thereof, wherein the linker has the following structure:

50. The compound of any one of claims 47-48, or a pharmaceutically acceptable salt thereof, wherein the linker has the following structure:

51. The compound of any one of claims 1 to 15 and 25 to 46, or a pharmaceutically acceptable salt thereof, wherein the linker has the structure of Formula III: A 1 -(B 1 ) f -(C 1 ) g -(B 2 ) h -(B 3 ) i -(C 2 ) j -(B 4 ) k –A 2 , Formula III in A 1 is the bond between the linker and the ring system A; A 2 is the bond between the degradable moiety and the linker; B 1 , B 2 , B 3 and B 4 are each independently an optionally substituted ethynyl, an optionally substituted C6-C 10 Aryl, optionally substituted C3-C 10 Cycloalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C10 heterocyclyl, optionally substituted C2-C9 heteroaryl, O, S, S(O)2 or NRN; Each RN is independently H, optionally substituted C 1–4 Alkyl, optionally substituted C 2–4 Alkenyl, optionally substituted C 2–4 Alkynyl, optionally substituted C 2–10 Heterocyclic group, optionally substituted C 6–12 Aryl or optionally substituted C 1–7 heteroalkyl; C 1 and C 2 are each independently carbonyl, thiocarbonyl, sulfonyl or phosphoryl; and f, g, h, i, j and k are each independently 0 or 1; B is the degradation part; each R1 is independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8 cycloalkyl, or optionally substituted C2-C10 heterocyclyl; and Each X is independently halogen.

52. The compound of claim 51 or a pharmaceutically acceptable salt thereof, wherein the linker has the following structure:

53. A pharmaceutical composition comprising a compound according to any one of claims 1 to 52 and a pharmaceutically acceptable excipient.

54. A method of treating a BAF complex-associated disorder in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of any one of claims 1 to 52 or a pharmaceutical composition of claim 53.

55. The method of claim 54, wherein the BAF complex-associated disorder is cancer or a viral infection.

56. A method of treating a disorder associated with a BRG1 loss-of-function mutation in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of any one of claims 1 to 52 or a pharmaceutical composition of claim 53.

57. The method of claim 56, wherein the disorder associated with a BRG1 loss-of-function mutation is cancer.

58. A method of treating cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of any one of claims 1 to 52 or a pharmaceutical composition of claim 53.

59. The method of any one of claims 54-58, wherein the cancer is non-small cell lung cancer, colorectal cancer, bladder cancer, cancer of unknown primary, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, esophageal-gastric cancer, pancreatic cancer, hepatobiliary cancer, soft tissue sarcoma, ovarian cancer, head and neck cancer, renal cell carcinoma, bone cancer, non-Hodgkin 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 intestine cancer, or penile cancer.

60. The method of claim 59, wherein the cancer is non-small cell lung cancer, colorectal cancer, bladder cancer, cancer of unknown primary, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, or penile cancer.

61. The method of claim 59, wherein the cancer is non-small cell lung cancer.

62. The method of claim 59, wherein the cancer is a soft tissue sarcoma.

63. A method of treating a cancer selected from melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, and a blood cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of any one of claims 1 to 52 or a pharmaceutical composition of claim 53.

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

65. A compound according to any one of claims 1 to 252, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 53, for use in the treatment of cancer.

66. A compound or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition for use according to claim 65, wherein the cancer is non-small cell lung cancer, colorectal cancer, bladder cancer, cancer of unknown primary, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, esophageal-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 intestine cancer or penile cancer.

67. A compound or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition for use according to claim 65, wherein the cancer is non-small cell lung cancer, colorectal cancer, bladder cancer, cancer of unknown primary focus, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer or penile cancer.

68. A compound or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition for use according to claim 65, wherein the cancer is non-small cell lung cancer.

69. A compound or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition for use according to claim 65, wherein the cancer is a soft tissue sarcoma.

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