Compounds and uses thereof

Abstract

The disclosure features compounds and methods useful for treating BAF complex-associated disorders.

Description

[0001] Background

[0002] The present invention relates to compounds that can be used to modulate BRG1 or BRM associated factor (BAF) complexes. In particular, the present invention relates to compounds that can be used to treat conditions associated with the function of BAF complexes.

[0003] Chromatin regulation is essential for gene expression, and ATP-dependent chromatin remodeling is the mechanism by which this gene expression occurs. The human SWItch / Sucrose Non-Fermentable (SWI / SNF) chromatin remodeling complex (also known as the BAF complex) has two SWI2-like ATPases called BRG1 (Brahma-related gene 1) and BRM (Brahma). The transcriptional activator BRG1 (also known as the ATP-dependent chromatin remodeler 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 putative global transcriptional activator SNF2L2 and / or ATP-dependent chromatin remodeler SMARCA2) is encoded by the SMARCA2 gene on chromosome 9 and has been shown to be essential for tumor cell growth in cells characterized by loss of BRG1 function mutations. Inactivation of BRG and / or BRM leads to downstream effects in cells, including cell cycle arrest and tumor suppression. Summary of the Invention

[0004] The present invention features compounds that can be used to modulate BAF complexes. In some embodiments, the compounds can be used to treat conditions associated with alterations in BAF complexes, such as conditions associated with alterations in one or both of the BRG1 and BRM proteins. The compounds of the present invention, alone or in combination with other pharmaceutically active agents, can be used to treat such conditions.

[0005] In one aspect, the present invention features compounds having a structure of Formula I:

[0006]

[0007] Wherein

[0008] L is a linker;

[0009] B is a degrading moiety; and

[0010] A has a structure of Formula II:

[0011]

[0012] Wherein

[0013] X 1 is N or CH,

[0014] X 2 and X 3 are independently N, CH or C(CH 3 );

[0015] R 1 is H, optionally substituted C 1 -C 6 acyl, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl, optionally substituted C 2 -C 9 heterocyclic group or -SO 2 R 6 ;

[0016] R 2 and R 5 are each independently H or optionally substituted C 1 -C 6 alkyl;

[0017] R 3 is H, optionally substituted C 1 -C 6 alkyl, or the bond between A and the linking group;

[0018] R 4 is H, optionally substituted C 1 -C 6 alkyl or optionally substituted C 1- C 6 heteroalkyl;

[0019] R 6 is optionally substituted C 1 -C 6 alkyl or -NR 7 R 8 ;

[0020] R 7 and R 8 are each independently optionally substituted C 1 -C 6 alkyl;

[0021] Het is a 5- or 6-membered heteroarylene group;

[0022] G 1 is optionally substituted C 6 -C 10 arylene, optionally substituted C 2 -C 9 heterocyclene or optionally substituted C 2- C 9Hetroarylene;

[0023] G 2 is absent, -O-, optionally substituted C 1 -C 6 alkylene, optionally substituted C 1 -C 6 alkenylene, optionally substituted C 1 -C 6 heteroalkylene, optionally substituted C 2 -C 9 heterocyclic C 1 -C 6 alkylene, or optionally substituted C 2 -C 9 heteroaryl C 1 -C 6 alkylene;

[0024] G 3 is absent, optionally substituted C 6 -C 10 arylene, optionally substituted C 6 -C 10 cycloalkylene, optionally substituted C 2 -C 9 heterocycloalkylene, or optionally substituted C 2 -C 9 heteroarylene;

[0025] and

[0026] A 1 is H or a bond between A and the linking group,

[0027] provided that formula II contains one and only one bond between A and the linking group,

[0028] or a pharmaceutically acceptable salt thereof.

[0029] In some embodiments, R 3 is H or optionally substituted C 1 -C6 alkyl, A 1 is a bond between A and the linking group. In some embodiments, R 3 is a bond between A and the linking group, A 1 is H.

[0030] In some embodiments, R 2 is hydrogen.

[0031] In some embodiments, R 5 is hydrogen. In other embodiments, R 5 is optionally substituted C 1 -C 6An alkyl group, such as a methyl group.

[0032] In some embodiments, X 1 , X 2 and X 3 are CH. In some embodiments, X 1 is N and X 2 and X 3 are CH. In some embodiments, X 3 is N and X 1 and X 2 are CH. In some embodiments, X 2 is N and X 1 and X 3 are CH. In some embodiments, X 1 is CH and X 2 and X 3 are C(CH 3 ). In some embodiments, X 1 and X 3 are CH and X 2 is C(CH 3 ).

[0033] In some embodiments, R 3 is hydrogen. In some embodiments, R 3 is an optionally substituted C 1 -C 6 alkyl group, such as R 3 is a methyl group.

[0034] In a further embodiment, R 4 is hydrogen. In other embodiments, R 4 is an optionally substituted C 1 -C 6 alkyl group, such as a methyl group, a tert-butyl group, an isopropyl group, an isobutyl group or a tert-pentyl group. In a further embodiment, R 4 is an optionally substituted C 1 -C 6 heteroalkyl group, such as .

[0035] In one embodiment, Het is . In other embodiments, Het is . In a further embodiment, Het is .

[0036] In some embodiments, G 2 is absent. In some embodiments, G 2 is an optionally substituted C 1 -C 6Alkylene, such as G 2 is 。In a further embodiment, G 2 is optionally substituted C 1 -C 6 heteroalkylene, optionally substituted C 2 -C 9 heterocyclylene or optionally substituted C 2 -C 9 heterocyclic C 1 -C 6 alkylene, such as

[0037]

[0038] 。In a particular embodiment, G 2 is optionally substituted C 1 -C 6 alkenylene, such as 。

[0039] In some embodiments, G 1 is optionally substituted C 6 -C 10 arylene, such as

[0040]

[0041] 。

[0042] In other embodiments, G 1 is optionally substituted C 2 -C 9 heteroarylene, such as

[0043]

[0044] 。

[0045] In a further embodiment, G 1 is optionally substituted C 2 -C 9 heterocyclylene, such as

[0046]

[0047]

[0048] 。

[0049] In some embodiments, G 3 is absent.

[0050] In further embodiments, G 3 is an optionally substituted C 6 -C 10 arylene, such as

[0051] .

[0052] In further embodiments, G 3 is an optionally substituted C 2 -C 9 heteroarylene, such as

[0053]

[0054]

[0055]

[0056] . In some embodiments, G 3 is wherein R 9a , R 9b , R 9c and R 9d are independently A 1 , H, halogen, hydroxy, an optionally substituted C 1-6 alkyl or an optionally substituted C 1-6 heteroalkyl; and R 9e and R 9f are each independently H or A 1 . In other embodiments, G 3 is wherein X 4 is O or CR 10i R 10j ; and R 10a , R 10b , R 10c , R 10d , R 10e , R 10f , R 10g , R 10h , R 10i and R 10j are each independently H, halogen, cyano, amino, hydroxy, allyl, heteroallyl, an optionally substituted C 1 - 6 alkyl, an optionally substituted C 1-6A heteroalkyl group, or two carbons attached thereto combine to form C=O.

[0057] In other embodiments, G 3 is an optionally substituted C 2 -C 9 heteroarylene, such as

[0058]

[0059]

[0060]

[0061] .

[0062] In a further embodiment, G 3 is an optionally substituted C 6 -C 10 cycloalkylene, such as

[0063]

[0064] .

[0065] In some embodiments, A 1 , G 1 , G 2 and G 3 combine to form an optionally substituted C 6 -C 10 aryl, such as

[0066]

[0067] .

[0068] In other embodiments, A 1 , G 1 , G 2 and G 3 combine to form an optionally substituted C 2 -C 9 heteroaryl, such as

[0069]

[0070] .

[0071] In a further embodiment, A 1 , G 1 , G 2 and G 3Combine to form an optionally substituted C 2 -C 9 heterocyclic group, such as

[0072]

[0073]

[0074] 。

[0075] In certain embodiments, R 1 is hydrogen. In other embodiments, R 1 is an optionally substituted C 1 -C 6 acyl group, such as acetyl. In a further embodiment, R 1 is an optionally substituted C 1 -C 6 alkyl group, such as methyl, tert-butyl, isopropyl or 。In other embodiments, R 1 is an optionally substituted C 1 -C 6 heteroalkyl group, such as 。In some embodiments, R 1 is an optionally substituted C 2 -C 9 heterocyclic group, such as 。

[0076] In a further embodiment, R 1 is –SO 2 R 6 。In some embodiments, R 6 is an optionally substituted C 1 -C 6 alkyl group, such as methyl, isopropyl or 。In some embodiments, R 1 is 。

[0077] In some embodiments, the degrading moiety is a ubiquitin ligase binding moiety.

[0078] In some embodiments, the ubiquitin ligase binding moiety comprises a Cereblon ligand, an IAP (inhibitor of apoptosis) ligand, a murine double minute 2 homolog (MDM2), or a von Hippel-Lindau (VHL) ligand, or a derivative or analogue thereof.

[0079] In some embodiments, the degrading moiety is a ubiquitin ligase binding moiety.

[0080] In some embodiments, the ubiquitin ligase binding moiety comprises a Cereblon ligand, an IAP (inhibitor of apoptosis) ligand, murine double minute 2 homolog (MDM2), or von Hippel-Lindau (VHL) ligand, or a derivative or analog thereof.

[0081] In some embodiments, the degrading moiety comprises a structure of formula Y:

[0082]

[0083] Wherein

[0084] A 2 is a bond between the degrading moiety and the linker;

[0085] v1 is 0, 1, 2, 3, 4 or 5;

[0086] u1 is 1, 2 or 3;

[0087] T 1 is a bond or ;

[0088] T 2 is

[0089] R 5A is H, optionally substituted C 1 -C 6 alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0090] R J1 are each independently halogen, optionally substituted C 1 -C 6 alkyl or optionally substituted C 1 -C 6 heteroalkyl; and

[0091] J is absent, optionally substituted C 3 -C 10 subcarbocyclene, optionally substituted C 6 -C 10 arylene, optionally substituted C 2 -C 9 heterocyclene or optionally substituted C 2 -C 9 heteroarylene or a pharmaceutically acceptable salt thereof.

[0092] In some embodiments, T 1 is a bond. In some embodiments, T 1 is .

[0093] In some embodiments, T 2 is 。In some embodiments, T 2 is 。In some embodiments, T 2 is 。

[0094] In some embodiments, the structure of formula Y has the structure of formula Y1:

[0095]

[0096] or a pharmaceutically acceptable salt thereof.

[0097] In some embodiments, the structure of formula Y has the structure of formula Y2:

[0098]

[0099] or a pharmaceutically acceptable salt thereof.

[0100] In some embodiments, the structure of formula Y has the structure of formula Z:

[0101]

[0102] or a pharmaceutically acceptable salt thereof.

[0103] In some embodiments, u1 is 1. In some embodiments, u1 is 2. In some embodiments, u1 is 3.

[0104] In some embodiments, the structure of formula Z has the structure of formula AA:

[0105]

[0106] or a pharmaceutically acceptable salt thereof.

[0107] In some embodiments, the structure of formula Z has the structure of formula AB:

[0108]

[0109] or a pharmaceutically acceptable salt thereof.

[0110] In some embodiments, the structure of formula Z has the structure of formula AC:

[0111]

[0112] or a pharmaceutically acceptable salt thereof.

[0113] In some embodiments, v1 is 0, 1, 2, or 3. In some embodiments, v1 is 0. In some embodiments, v1 is 1. In some embodiments, v1 is 2. In some embodiments, v1 is 3.

[0114] In some embodiments, the structure of formula AA has the structure of formula AA1:

[0115]

[0116] or a pharmaceutically acceptable salt thereof.

[0117] In some embodiments, the structure of formula AB has the structure of formula AB1:

[0118]

[0119] or a pharmaceutically acceptable salt thereof.

[0120] In some embodiments, the structure of formula AC has the structure of formula AC1:

[0121]

[0122] or a pharmaceutically acceptable salt thereof.

[0123] In some embodiments, J is absent. In some embodiments, J is an optionally substituted C 3 -C 10 subcarbocyclic group or an optionally substituted C 6 -C 10 subaryl group. In some embodiments, J is an optionally substituted C 2 -C 9 subheterocyclic group or an optionally substituted C 2 -C 9 subheteroaryl group.

[0124] In some embodiments, J is an optionally substituted subheterocyclic group. In some embodiments, J is an optionally substituted C 6 -C 10 subaryl group.

[0125] In some embodiments, the structure of formula AA has the structure of formula AA2:

[0126]

[0127] or a pharmaceutically acceptable salt thereof.

[0128] In some embodiments, the structure of formula AA has the structure of formula AA3:

[0129]

[0130] or a pharmaceutically acceptable salt thereof.

[0131] In some embodiments, the structure of formula AA has the structure of formula AA4:

[0132]

[0133] or a pharmaceutically acceptable salt thereof.

[0134] In some embodiments, R A5 is H or optionally substituted C 1 -C 6 alkyl. In some embodiments, R A5 is H or methyl. In some embodiments, R A5 is H. In some embodiments, R A5 is methyl.

[0135] In some embodiments, the structure of formula AA has the structure of formula A:

[0136]

[0137] wherein

[0138] Y 1 is

[0139] R A5 is H, optionally substituted C 1 -C 6 alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0140] 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 together with the respective carbon atoms to which they are attached form an optionally substituted C 3- C 6 carbocyclic group or an optionally substituted C 2 -C 5 heterocyclic group; or R A6 and R A7 together with the respective carbon atoms to which they are attached form an optionally substituted C 3 -C 6 carbocyclic group or an optionally substituted C 2 -C 5 heterocyclic group;

[0141] R A8 is H, optionally substituted C 1 -C 6 alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0142] R A1 、R A2 、R A3 and R A4 are each independently 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 -O-C 3 -C 6 carbocyclic group, hydroxyl, mercapto or optionally substituted amino; or R A1 and R A2 、R A2 and R A3 、and / or R A3 and R A4 together with the carbon atom to which they are attached form ; and is 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 substituted by A 2 substituted, wherein R A1 、R A2 、R A3 and R A4 one of which is A 2 ,or is substituted by A 2 , or a pharmaceutically acceptable salt thereof.

[0143] In some embodiments, R A1 , R A2 , R A3 and R A4 are each independently 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, hydroxyl, mercapto or optionally substituted amino; or R A1 and R A2 , R A2 and R A3 , and / or R A3 and R A4 combine with the carbon atom to which they are attached to form ; and is 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 substituted by A 2 , wherein one of R A1 , R A2 , R A3 and R A4 is A 2 , or is substituted by A 2 , or a pharmaceutically acceptable salt thereof.

[0144] In some embodiments, R A1 , R A2 , R A3 and R A4 are each H, A 2 , halogen, optionally substituted C1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl, optionally substituted -O-C 3 -C 6 carbocyclic group, hydroxyl group, optionally substituted amino group; or R A1 and R A2 , R A2 and R A3 , or R A3 and R A4 combine with the respective connected carbon atoms to form ; and is an optionally substituted C 2 -C 9 heterocyclic group, which is optionally substituted by A 2 , where R A1 , R A2 , R A3 and R A4 one of them is A 2 , or is substituted by A 2 .

[0145] In some embodiments, R A1 , R A2 , R A3 and R A4 are each independently H, A 2 , F, or R A1 and R A2 , R A2 and R A3 , or R A3 and R A4 combine with the respective connected carbon atoms to form ; and is an optionally substituted C 2 -C 9 heterocyclic group, which is optionally substituted by A 2 , where R A1 , R A2 , R A3 and R A4 one of them is A 2 , or is substituted by A 2 .

[0146] In some embodiments, R A1 is A 2 . In some embodiments, R A2 is A 2 . In some embodiments, RA3 is A 2 . In some embodiments, R A4 is A 2 . In some embodiments, R A5 is A 2 .

[0147] In some embodiments, R A5 is H or optionally substituted C 1 -C 6 alkyl.

[0148] In some embodiments, R A5 is H or . In some embodiments, R A5 is H. In some embodiments, R A5 is .

[0149] In some embodiments, Y 1 is . In some embodiments, Y 1 is . In some embodiments, Y 1 is .

[0150] In some embodiments, R A6 and R A7 are each independently H, F, ; or R A6 and R A7 combine with the carbon atom to which they are attached to form .

[0151] In some embodiments, Y 1 is .

[0152] In some embodiments, the structure of Formula A has the structure of Formula A1:

[0153]

[0154] or a pharmaceutically acceptable salt thereof.

[0155] In some embodiments, the structure of Formula A has the structure of Formula A2:

[0156]

[0157] or a pharmaceutically acceptable salt thereof.

[0158] In some embodiments, the structure of Formula A has the structure of Formula A3:

[0159]

[0160] or a pharmaceutically acceptable salt thereof.

[0161] In some embodiments, the structure of formula A has the structure of formula A4:

[0162]

[0163] or a pharmaceutically acceptable salt thereof.

[0164] In some embodiments, the structure of formula A has the structure of formula A5:

[0165]

[0166] or a pharmaceutically acceptable salt thereof.

[0167] In some embodiments, the structure of formula A has the structure of formula A6:

[0168]

[0169] or a pharmaceutically acceptable salt thereof.

[0170] In some embodiments, the structure of formula A has the structure of formula A7:

[0171]

[0172] or a pharmaceutically acceptable salt thereof.

[0173] In some embodiments, the structure of formula A has the structure of formula A8:

[0174]

[0175] or a pharmaceutically acceptable salt thereof.

[0176] In some embodiments, the structure of formula A has the structure of formula A9:

[0177]

[0178] or a pharmaceutically acceptable salt thereof.

[0179] In some embodiments, the structure of formula A has the structure of formula A10:

[0180]

[0181] or a pharmaceutically acceptable salt thereof.

[0182] In some embodiments, wherein the structure of formula A is

[0183]

[0184]

[0185] , or its derivatives or analogs.

[0186] In some embodiments, the structure of Formula A is

[0187] .

[0188] In some embodiments, the structure of Formula A is

[0189] , or its derivatives or analogs.

[0190] In some embodiments, is , where R A9 is H, A 2 , optionally substituted C 1 -C 6 alkyl or optionally substituted C 1 -C 6 heteroalkyl.

[0191] In some embodiments, the structure of Formula A is

[0192]

[0193] .

[0194] In some embodiments, R A9 is H, A 2 or optionally substituted C 1 -C 6 alkyl. In some embodiments, R A9 is H, A 2 or methyl. In some embodiments, R 9A is H. In some embodiments, R 9A is methyl. In some embodiments, R A9 is A 2 .

[0195] In some embodiments, the structure of Formula A is

[0196] .

[0197] In some embodiments, the structure of formula AA has the structure of formula B:

[0198]

[0199] where

[0200] R A5 is H, optionally substituted C 1 -C 6 alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0201] R A1 、R A2 、R A3 and R A4 are each independently 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 -O-C 3 -C 6 carbocyclic group, hydroxy, mercapto or optionally substituted amino; or R A1 and R A2 、R A2 and R A3 、and / or R A3 and R A4 combine with the respective connected carbon atoms to form ; and is 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 substituted by A 2 substituted, where RA1 , R A2 , R A3 and R A4 One of them is A 2 , or is replaced by A 2 , or a pharmaceutically acceptable salt thereof.

[0202] In some embodiments, R A1 , R A2 , R A3 and R A4 are each H, A 2 , halogen, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl, optionally substituted -O-C 3 -C 6 carbocyclic group, hydroxy, optionally substituted amino; or R A1 and R A2 , R A2 and R A3 , or R A3 and R A4 combine with the carbon atom to which they are attached to form ; and is an optionally substituted C 2 -C 9 heterocyclic group, optionally substituted by A 2 , where R A1 , R A2 , R A3 and R A4 One of them is A 2 , or is replaced by A 2 .

[0203] In some embodiments, R A1 , R A2 , R A3 and R A4 are each independently H, A 2 , F, ; or R A1 and R A2 , R A2 and R A3 , or R A3 and R A4 combine with the carbon atom to which they are attached to form ; and is an optionally substituted C 2 -C 9 heterocyclic group, optionally substituted by A2 is substituted, wherein R A1 , R A2 , R A3 and R A4 is one of A 2 , or is substituted by A 2 .

[0204] In some embodiments, R A1 is A 2 . In some embodiments, R A2 is A 2 . In some embodiments, R A3 is A 2 . In some embodiments, R A4 is A 2 . In some embodiments, R A5 is A 2 .

[0205] In some embodiments, R A5 is H or optionally substituted C 1 -C 6 alkyl.

[0206] In some embodiments, R A5 is H or . In some embodiments, R A5 is H. In some embodiments, R A5 is .

[0207] In some embodiments, the structure of formula B has the structure of formula B1:

[0208]

[0209] or a pharmaceutically acceptable salt thereof.

[0210] In some embodiments, the structure of formula B has the structure of formula B2:

[0211]

[0212] or a pharmaceutically acceptable salt thereof.

[0213] In some embodiments, the structure of formula B has the structure of formula B3:

[0214]

[0215] or a pharmaceutically acceptable salt thereof.

[0216] In some embodiments, the structure of formula B has the structure of formula B4:

[0217]

[0218] or a pharmaceutically acceptable salt thereof.

[0219] In some embodiments, the structure of Formula B is

[0220] . In some embodiments, the structure of Formula B is . In some embodiments, the structure of Formula B is .

[0221] In some embodiments, the ubiquitin ligase binding moiety comprises a von Hippel-Lindau ligand.

[0222] In some embodiments, the von Hippel-Lindau ligand has the following structure: ,

[0223] or a derivative or analog thereof.

[0224] In some embodiments, the degrading moiety comprises a structure of Formula C:

[0225]

[0226] wherein

[0227] R B1 and R B9 are independently H, A 2 , optionally substituted C 1 -C 6 alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0228] R B2 is H, optionally substituted C 1 -C 6 alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0229] R B3 is A 2 , optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl, optionally substituted C 3 -C 10 carbocyclic, 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;

[0230] R B4 is H, optionally substituted C 1 -C 6 alkyl, optionally substituted C 3 -C 10 carbocyclic, 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;

[0231] R B5 is H, optionally substituted C 1 -C 6 alkyl or optionally substituted C 1 -C 6 heteroalkyl;

[0232] v2 is 0, 1, 2, 3 or 4;

[0233] R B6 each independently is halogen, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl, optionally substituted C 3 -C 10 carbocyclic, optionally substituted C 2 -C 9 heterocyclic, 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, mercapto or optionally substituted amino; and

[0234] R B7 and R B8Each independently is H, a halogen, an optionally substituted C 1 -C 6 alkyl or an optionally substituted C 6 -C 10 aryl,

[0235] wherein R B1 、R B3 、R B6 and R B9 One of them is A 2 , or a pharmaceutically acceptable salt thereof.

[0236] In some embodiments, the structure of Formula C is

[0237] , or a derivative or analog thereof. In some embodiments, the structure of Formula C is . In some embodiments, the structure of Formula C is

[0238] . In some embodiments, the structure of Formula C is

[0239] .

[0240] In some embodiments, the degradant moiety is the degradant moiety described in International Patent Publication WO2019 / 195201, which is incorporated herein by reference.

[0241] In some embodiments, the degradant moiety comprises a structure of Formula D:

[0242]

[0243] wherein

[0244] A 2 is a bond between B and the linker;

[0245] R C1 、R C2 and R C7 Each independently is H, an optionally substituted C 1 -C 6 alkyl or an optionally substituted C 1 -C 6 heteroalkyl;

[0246] R C3 is an optionally substituted C 1 -C 6 alkyl, an optionally substituted C 3 -C 10 carbocyclic group, an optionally substituted C 6 -C10 Aryl, optionally substituted C 1 -C 6 alkyl C 3 -C 10 carbocyclic group or optionally substituted C 1 -C 6 alkyl C 6 -C 10 aryl;

[0247] R C5 is 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 group or optionally substituted C 1 -C 6 alkyl C 6 -C 10 aryl;

[0248] v3 is 0, 1, 2, 3 or 4;

[0249] R C8 each independently is 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, mercapto or optionally substituted amino;

[0250] v4 is 0, 1, 2, 3 or 4; and

[0251] R C9 each independently is halogen, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C6 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, hydroxyl, mercapto or optionally substituted amino, or a pharmaceutically acceptable salt thereof.

[0252] In some embodiments, the structure of formula D is , or a derivative or analogue thereof.

[0253] In some embodiments, the degradant moiety comprises a structure of formula E:

[0254]

[0255] Wherein

[0256] A 2 is a bond between B and the linker;

[0257] R C10 and R C11 are each independently 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 group or optionally substituted C 1 -C 6 alkyl C 6 -C 10 aryl;

[0258] v5 is 0, 1, 2, 3 or 4;

[0259] R C12 are each independently halogen, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl, optionally substituted C 3 -C10 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, hydroxyl, mercapto or optionally substituted amino

[0260] v6 is 0, 1, 2, 3 or 4; and

[0261] R 21 Each independently is 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, hydroxyl, mercapto or optionally substituted amino, or a pharmaceutically acceptable salt thereof.

[0262] In some embodiments, the structure of Formula E is , or a derivative or analogue thereof.

[0263] In some embodiments, the degradation moiety comprises a structure of Formula FA:

[0264]

[0265] Wherein

[0266] is , or a bicyclic moiety substituted by A 2 and substituted by one or more groups independently selected from H, R FF1 and oxo;

[0267] is a single bond or a double bond;

[0268] u2 is 0, 1, 2 or 3;

[0269] A 2 is the bond between the degrading substance and the linker;

[0270] Y Fa is CR Fb R Fc 、C═O、C═S、C═CH 2 、SO 2 、S(O)、P(O)O alkyl, P(O)NH alkyl, P(O)N(alkyl) 2 、P(O) alkyl, P(O)OH, P(O)NH 2 ;

[0271] Y Fb is NH, NR FF1 、CH 2 、CHR FF1 、C(R FF1 ) 2 、O or S;

[0272] Y Fc is CR Fd R Fe 、C═O、C═S、C═CH 2 、SO 2 、S(O)、P(O)O alkyl, P(O)NH alkyl, P(O)N(alkyl) 2 、P(O) alkyl, P(O)OH, P(O)NH 2 ;

[0273] R Fb 、R Fc 、R Fd and R Fe are each independently H, alkyl, aliphatic group, heteroaliphatic group, aryl, heteroaryl, carbocyclic group, hydroxy, alkoxy, amino, —NH alkyl, or —N alkyl 2 ;

[0274] or R Fb and R Fc together with the carbon atoms to which they are attached form a 3-, 4-, 5- or 6-membered spirocarbocyclic group, or a 4-, 5- or 6-membered spiroheterocyclic group containing 1 or 2 heteroatoms selected from N and O;

[0275] or R Fd and R Fe together with the carbon atoms to which they are attached form a 3-, 4-, 5- or 6-membered spirocarbocyclic group, or a 4-, 5- or 6-membered spiroheterocyclic group containing 1 or 2 heteroatoms selected from N and O;

[0276] or R Fd and R Fb combine with the respective attached carbon atoms to form a 1, 2, 3 or 4 carbon bridged ring;

[0277] Y Fd and Y Ff are each independently CH 2 , CHR FF2 , C(R FF2 ) 2 , C(O), N, NH, NR FF3 , O, S or S(O);

[0278] Y Fe is a bond or a divalent group attached to Y Fd and Y Ff and contains 1 to 5 connected carbon atoms forming a 3 to 8-membered ring,

[0279] where 1, 2 or 3 carbon atoms may be replaced by nitrogen, oxygen or sulfur atoms;

[0280] where one of the ring atoms is substituted by A 2 and the other ring atoms are substituted by one or more groups independently selected from H and R FF1 ; and

[0281] where Y Fe 's connected atoms can be connected by single or double bonds;

[0282] R FF1 are each independently H, alkyl, alkenyl, alkynyl, aliphatic group, heteroaliphatic group, carbocyclic group, halogen, hydroxy, amino, cyano, alkoxy, aryl, heteroaryl, heterocyclic group, alkylamino, alkylhydroxy or haloalkyl;

[0283] R FF2 are each independently alkyl, alkenyl, alkynyl, halogen, hydroxy, alkoxy, azido, amino,

[0284] —C(O)H, —C(O)OH, —C(O)(aliphatic group, including alkyl), —C(O)O(aliphatic group, including alkyl),

[0285] —NH(aliphatic group, including alkyl), —N(aliphatic group, including alkyl)(aliphatic group, including alkyl), —NHSO 2 alkyl,

[0286] —N(alkyl)SO 2 alkyl, —NHSO 2 aryl, —N(alkyl)SO 2 aryl, —NHSO2 alkenyl, —N(alkyl)SO 2 alkenyl,

[0287] —NHSO 2 alkynyl, —N(alkyl)SO 2 alkynyl, aliphatic group, heteroaliphatic group, aryl, heteroaryl, heterocyclic group, carbocyclic group, cyano, nitro, nitroso, —SH —S alkyl or haloalkyl; and

[0288] R FF3 is alkyl, alkenyl, alkynyl, —C(O)H, —C(O)OH, —C(O)alkyl or —C(O)Oalkyl,

[0289] wherein if Y Fd or Y Ff is substituted by A 2 then Y Fe is a bond, or a pharmaceutically acceptable salt thereof.

[0290] In some embodiments, the compound of formula FA has the structure of formula FA1:

[0291]

[0292] or a pharmaceutically acceptable salt thereof.

[0293] In some embodiments, the degradable moiety comprises a structure of formula FB:

[0294]

[0295] wherein

[0296] is or a bicyclic moiety substituted by A 2 and substituted by one or more groups independently selected from H, R FF1 and oxo;

[0297] A 2 is the bond between the degradant and the linker;

[0298] Y Fa is CR Fb R Fc 、C═O、C═S、C═CH 2 、SO 2 、S(O)、P(O)Oalkyl、P(O)NHalkyl、P(O)N(alkyl) 2 、P(O)alkyl、P(O)OH, P(O)NH 2 ;

[0299] Y Fb and YFg each independently is NH, NR FF1 , CH 2 , CHR FF1 , C(R FF1 ) 2 , O or S;

[0300] Y Fc is CR Fd R Fe , C═O, C═S, C═CH 2 , SO 2 , S(O), P(O)O alkyl, P(O)NH alkyl, P(O)N(alkyl) 2 , P(O) alkyl, P(O)OH, P(O)NH 2 ;

[0301] R Fb , R Fc , R Fd , R Fe , R Ff and R Fg each independently is H, alkyl, aliphatic group, heteroaliphatic group, aryl, heteroaryl, carbocyclic group, hydroxy, alkoxy, amino, —NH alkyl or —N alkyl 2 ;

[0302] or R Fb and R Fc together with the carbon atoms to which they are attached form a 3-, 4-, 5- or 6-membered spirocarbocyclic group, or a 4-, 5- or 6-membered spiroheterocyclic group containing 1 or 2 heteroatoms selected from N and O;

[0303] or R Fd and R Fe together with the carbon atoms to which they are attached form a 3-, 4-, 5- or 6-membered spirocarbocyclic group, or a 4-, 5- or 6-membered spiroheterocyclic group containing 1 or 2 heteroatoms selected from N and O;

[0304] or R Ff and R Fg together with the carbon atoms to which they are attached form a 3-, 4-, 5- or 6-membered spirocarbocyclic group, or a 4-, 5- or 6-membered spiroheterocyclic group containing 1 or 2 heteroatoms selected from N and O;

[0305] or R Fd and R Fb together with the carbon atoms to which they are attached form a 1, 2, 3 or 4 carbon bridged ring;

[0306] or R Fd and R FfCombine with the attached carbon atoms respectively to form a 1, 2, 3 or 4-carbon bridged ring;

[0307] or R Fd and R Fg Combine with the attached carbon atoms respectively to form a 1, 2, 3 or 4-carbon bridged ring;

[0308] Y Fd and Y Ff are each independently CH 2 、CHR FF2 、C(R FF2 ) 2 、C(O), N, NH, NR FF3 、O, S or S(O);

[0309] Y Fe is a bond or a divalent group attached to Y Fd and Y Ff and contains 1 to 5 connected carbon atoms forming a 3- to 8-membered ring

[0310] wherein 1, 2 or 3 carbon atoms may be replaced by nitrogen, oxygen or sulfur atoms;

[0311] wherein one of the ring atoms is replaced by A 2 and the other ring atoms are replaced by one or more groups independently selected from H and R FF1 ; and

[0312] wherein the connected atoms of Y Fe can be connected by a single bond or a double bond;

[0313] R FF1 are each independently H, alkyl, alkenyl, alkynyl, aliphatic group, heteroaliphatic group, carbocyclic group, halogen, hydroxy, amino, cyano, alkoxy, aryl, heteroaryl, heterocyclic group, alkylamino, alkylhydroxy or haloalkyl;

[0314] R FF2 are each independently alkyl, alkenyl, alkynyl, halogen, hydroxy, alkoxy, azido, amino,

[0315] —C(O)H, —C(O)OH, —C(O)(aliphatic group, including alkyl), —C(O)O(aliphatic group, including alkyl),

[0316] —NH(aliphatic group, including alkyl), —N(aliphatic group, including alkyl)(aliphatic group, including alkyl), —NHSO 2 alkyl,

[0317] —N(alkyl)SO 2 alkyl, —NHSO 2Aryl, —N(alkyl)SO 2 Aryl, —NHSO 2 Alkenyl, —N(alkyl)SO 2 Alkenyl,

[0318] —NHSO 2 Alkynyl, —N(alkyl)SO 2 Alkynyl, aliphatic group, heteroaliphatic group, aryl, heteroaryl, heterocyclic group, carbocyclic group, cyano, nitro, nitroso, —SH, —S-alkyl or haloalkyl; and

[0319] R FF3 is alkyl, alkenyl, alkynyl, —C(O)H, —C(O)OH, —C(O)alkyl or —C(O)Oalkyl,

[0320] wherein if Y Fd and Y Ff are substituted by A 2 then Y Fe is a bond, or a pharmaceutically acceptable salt thereof.

[0321] In some embodiments, the compound of formula FB has the structure of formula FB1:

[0322]

[0323] or a pharmaceutically acceptable salt thereof.

[0324] In some embodiments, the degradable moiety comprises the structure of formula F1:

[0325]

[0326] wherein A 2 is the bond between the degradant and the linker; and R F1 is absent or O, or a pharmaceutically acceptable salt thereof.

[0327] In some embodiments, R F1 is absent. In some embodiments, R F1 is O.

[0328] In some embodiments, the structure of formula F1 is .

[0329] In some embodiments, the degradable moiety comprises the structure of formula F2:

[0330]

[0331] wherein A 2 is the bond between the degradant and the linker; and Y 2 is CH2 or NH, or a pharmaceutically acceptable salt thereof.

[0332] In some embodiments, Y 2 is NH. In some embodiments, Y 2 is CH 2 .

[0333] In some embodiments, the structure of Formula F2 is .

[0334] In some embodiments, the degradation moiety comprises a structure of Formula G:

[0335]

[0336] wherein A 2 is the bond between the degradant and the linker; and Y 3 is CH 2 or NH, or a pharmaceutically acceptable salt thereof.

[0337] In some embodiments, Y 3 is NH. In some embodiments, Y 3 is CH 2 .

[0338] In some embodiments, the structure of Formula G is .

[0339] The degradation moiety may also include, for example, the structures found in WO2017 / 197036, WO2019 / 204354, WO2019 / 236483, WO2020 / 010177, and WO2020 / 010227, each of which is incorporated herein by reference in its entirety.

[0340] In some embodiments, the linker has a structure of Formula III:

[0341]

[0342] wherein A 1 is the bond between A and the linker; A 2 is the bond between the linker and B; B 1 , B 2 , B 3 and B 4 are each independently an optionally substituted C 1 -C 2 alkyl, an optionally substituted C 1 -C 2 heteroalkyl, O, S, S(O) 2 or NR N ; R NEach is independently H, optionally substituted C 1–4 alkyl, optionally substituted C 2–4 alkenyl, optionally substituted C 2–4 alkynyl, optionally substituted C 2–6 heterocyclic group, optionally substituted C 6–12 aryl or optionally substituted C 1–7 heteroalkyl; C 1 and C 2 Each is independently a carbonyl, thiocarbonyl, sulfonyl or phosphoryl group; f, g, h, i, j and k are each independently 0 or 1; and D is optionally substituted C 1–12 alkyl, optionally substituted C 2–12 alkenyl, optionally substituted C 2–12 alkynyl, optionally substituted C 2 -C 12 polyethylene glycol or optionally substituted C 1–12 heteroalkyl, or a bond connecting A 1 -(B 1 ) f -(C 1 ) g -(B 2 ) h - and -(B 3 ) i -(C 2 ) j -(B 4 ) k –A 2 chemical bond.

[0343] In some embodiments, B 1 , B 2 , B 3 and B 4 are each independently optionally substituted C 1 -C 4 alkyl, optionally substituted C 1 -C 4 heteroalkyl or NR N .

[0344] In some embodiments, R N are each independently H or optionally substituted C 1 -C 4 alkyl.

[0345] In some embodiments, R N are each independently H or methyl.

[0346] In some embodiments, B 1 and B 4 are each independently

[0347] 。

[0348] In some embodiments, B 1 is 。

[0349] In some embodiments, C 1 and C 2 are each independently 。

[0350] In some embodiments, C 1 is 。

[0351] In some embodiments, B 2 is NR N 。In some embodiments, B 2 is optionally substituted C 1 -C 4 alkyl.

[0352] In some embodiments, f is 0. In some embodiments, f is 1. 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, j is 0. In some embodiments, k is 0.

[0353] In some embodiments, D is optionally substituted C 1–12 alkyl, optionally substituted C 2 -C 12 polyethylene glycol or optionally substituted C 1–12 heteroalkyl. In some embodiments, D is a chemical bond connecting A 1 -(B 1 ) f -(C 1 ) g -(B 2 ) h -and -(B 3 ) i -(C 2 ) j -(B 4 ) k –A 2 。

[0354] In some embodiments, the linking group has the following structure:

[0355]

[0356]

[0357] wherein x1 is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14; x2 is 0, 1, 2, 3, 4, 5 or 6; x3 is 1 or 2; x4 is 1 or 2; W is ; R x1 and R x2 are each independently H, halogen, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl or optionally substituted C 3 -C 6 carbocyclic group, or R x1 and R x2 together with the carbon atom to which each is attached combine to form an optionally substituted C 3 -C 6 carbocyclic group or optionally substituted C 2 -C 5 heterocyclic group; and R y1 , R y2 , R y3 and R y4 are each independently H, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl or optionally substituted C 3 -C 6 carbocyclic group.

[0358] In some embodiments, W is . In some embodiments, W is . In some embodiments, W is .

[0359] In some embodiments, R x1 and R x2 are each independently H, halogen, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl or optionally substituted C 3 -C 6 carbocyclic group.

[0360] In some embodiments, R x1 and R x2 are each independently H or optionally substituted C 1 -C 6 alkyl.

[0361] In some embodiments, R x1 and R x2 are each independently H or methyl.

[0362] In some embodiments, R x1 and R x2 combine with the carbon atoms to which they are attached to form an optionally substituted C 3 -C 6 carbocyclic group or an optionally substituted C 2 -C 5 heterocyclic group.

[0363] In some embodiments, R x1 and R x2 combine with the carbon atoms to which they are attached to form an optionally substituted C 3 -C 6 carbocyclic group.

[0364] In some embodiments, R x1 and R x2 combine with the carbon atoms to which they are attached to form cyclopropyl.

[0365] In some embodiments, R y1 , R y2 , R y3 and R y4 are each independently H or an optionally substituted C 1 -C 6 alkyl group.

[0366] In some embodiments, R y1 , R y2 , R y3 and R y4 are each independently H or methyl.

[0367] In some embodiments, the linking group has the following structure:

[0368]

[0369]

[0370]

[0371]

[0372]

[0373] .

[0374] In some embodiments, the linking group has the structure of Formula IV:

[0375]

[0376] wherein A 1 is the bond between the linking group and A; A 2 is the bond between B and the linking group; m1, m2, n1, n2, o1, o2, p1, and p2 are each independently 0 or 1; E 1 and E 2 are each independently O, S, NR N , optionally substituted C 1–10 alkyl, optionally substituted C 2–10 alkenyl, optionally substituted C 2–10 alkynyl, optionally substituted C 2 -C 10 polyethylene glycol, or optionally substituted C 1–10 heteroalkyl; E 3 are each independently optionally substituted C 1–10 alkyl, optionally substituted C 1–10 heteroalkyl, O, S, or NR N ; R N are each 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 3 and C 4 are each independently carbonyl, thiocarbonyl, sulfonyl, or phosphoryl; F 1 , F 2 and F 3 are each independently optionally substituted C 3 -C 10 carbocyclic group, optionally substituted C 2–10 heterocyclic group, optionally substituted C 6 -C 10 aryl, or optionally substituted C 2 -C 9 heteroaryl.

[0377] In some embodiments, the linking group has the structure of Formula IVa:

[0378] .

[0379] In some embodiments, the linking group has the structure of Formula IVb:

[0380] .

[0381] In some embodiments, the linking group has the structure of formula IVc:

[0382] .

[0383] In some embodiments, the linking group has the structure of formula IVd:

[0384] .

[0385] In some embodiments, the linking group has the structure of formula IVe:

[0386] .

[0387] In some embodiments, the linking group has the structure of formula IVf:

[0388] ,

[0389] wherein A 1 is the bond between the linking group and A; A 2 is the bond between B and the linking group; m1, m2, n1, o1, o2, p1, and p2 are each independently 0 or 1; E 1 and E 2 are each independently O, S, NR N , optionally substituted C 1–10 alkyl, optionally substituted C 2–10 alkenyl, optionally substituted C 2–10 alkynyl, optionally substituted C 2 -C 10 polyethylene glycol, or optionally substituted C 1–10 heteroalkyl; E 3 is independently optionally substituted C 1–2 alkyl, optionally substituted C 1–2 heteroalkyl, O, S, or NR N ; R N are each 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 3 and C 4 are each independently carbonyl, thiocarbonyl, sulfonyl, or phosphoryl; F 1 , F 2 and F 3 are each independently optionally substituted C 3 -C10 Carbocyclic group, optionally substituted C 2–10 heterocyclic group, optionally substituted C 6 -C 10 aryl or optionally substituted C 2 -C 9 heteroaryl.

[0390] In some embodiments, R N each independently is H or optionally substituted C 1–4 alkyl.

[0391] In some embodiments, R N each independently is H or methyl.

[0392] In some embodiments, E 1 and E 2 each independently is NR N , optionally substituted C 1–10 alkyl, optionally substituted C 2 -C 10 polyethylene glycol or optionally substituted C 1–10 heteroalkyl.

[0393] In some embodiments, E 1 and E 2 each independently is optionally substituted C 1–10 alkyl or optionally substituted C 1–10 heteroalkyl.

[0394] In some embodiments, E 1 is

[0395] , where z1 is 0, 1 or 2; z2 is 0, 1, 2, 3, 4, 5 or 6; R z1 and R z2 each independently is H, halogen, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl or optionally substituted C 3 -C 6 carbocyclic group, or R x1 and R x2 together with the carbon atom to which each is attached combine to form an optionally substituted C 3 -C 6 carbocyclic group or optionally substituted C 2 -C 5 heterocyclic group; and R a and R b each independently is H, optionally substituted C1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl or optionally substituted C 3 -C 6 carbocyclic group.

[0396] In some embodiments, z1 is 0. In some embodiments, z1 is 1.

[0397] In some embodiments, R z1 and R z2 are each independently H, halogen, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl or optionally substituted C 3 -C 6 carbocyclic group.

[0398] In some embodiments, R z1 and R z2 are each independently H or optionally substituted C 1 -C 6 alkyl.

[0399] In some embodiments, R z1 and R z2 are each independently H or methyl.

[0400] In some embodiments, R z1 and R z2 together with the carbon atom to which each is attached form an optionally substituted C 3 -C 6 carbocyclic group or an optionally substituted C 2 -C 5 heterocyclic group.

[0401] In some embodiments, R z1 and R z2 together with the carbon atom to which each is attached form an optionally substituted C 3 -C 6 carbocyclic group.

[0402] In some embodiments, R z1 and R z2 together with the carbon atom to which each is attached form cyclopropyl.

[0403] In some embodiments, R a and R b are each independently H or optionally substituted C 1 -C 6Alkyl.

[0404] In some embodiments, R a and R b are each independently H or methyl.

[0405] In some embodiments, E 2 is

[0406]

[0407]

[0408] , where z3 is 0, 1, 2, 3, 4, 5 or 6; z4 is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12; z5 is 0, 1, 2, 3 or 4; z6 is 1, 2, 3 or 4; R z3 and R z4 are each independently H, halogen, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl or optionally substituted C 3 -C 6 carbocyclic group, or R x1 and R x2 together with the carbon atom to which each is attached combine to form an optionally substituted C 3 -C 6 carbocyclic group or optionally substituted C 2 -C 5 heterocyclic group; and R c is H, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl or optionally substituted C 3 -C 6 carbocyclic group.

[0409] In some embodiments, z3 is 0, 1, 2 or 3.

[0410] In some embodiments, z4 is 0, 1, 2, 3, 4, 5 or 6.

[0411] In some embodiments, z5 is 0, 1 or 2.

[0412] In some embodiments, z6 is 1 or 2.

[0413] In some embodiments, R z3 and R z4Each is independently H, a halogen, an optionally substituted C 1 -C 6 alkyl, an optionally substituted C 1 -C 6 heteroalkyl or an optionally substituted C 3 -C 6 carbocyclic group.

[0414] In some embodiments, R z3 and R z4 are each independently H or an optionally substituted C 1 -C 6 alkyl.

[0415] In some embodiments, R z3 and R z4 are each independently H or methyl.

[0416] In some embodiments, R z3 and R z4 together with the carbon atom to which each is attached combine to form an optionally substituted C 3 -C 6 carbocyclic group or an optionally substituted C 2 -C 5 heterocyclic group.

[0417] In some embodiments, R z3 and R z4 together with the carbon atom to which each is attached combine to form an optionally substituted C 3 -C 6 carbocyclic group.

[0418] In some embodiments, R z3 and R z4 together with the carbon atom to which each is attached combine to form cyclopropyl.

[0419] In some embodiments, R c is H or an optionally substituted C 1 -C 6 alkyl.

[0420] In some embodiments, R c is H or methyl.

[0421] In some embodiments, E 3 is an optionally substituted C 1–2 alkyl, an optionally substituted C 1–2 heteroalkyl. In some embodiments, E 3 is O, S or NR N .

[0422] In some embodiments, E 3is an optionally substituted C 1–2 alkyl group.

[0423] In some embodiments, E 3 is .

[0424] In some embodiments, F 1 , F 2 or F 3 is each independently an optionally substituted C 3 -C 10 carbocyclic group. In some embodiments, the C 3 -C 10 carbocyclic group is monocyclic. In some embodiments, the C 3 -C 10 carbocyclic group is polycyclic. In some embodiments, the C 3 -C 10 carbocyclic group is bicyclic. In some embodiments, the C 3 -C 10 carbocyclic group is bridged. In some embodiments, the C 3 -C 10 carbocyclic group is fused. In some embodiments, the C 3 -C 10 carbocyclic group is spirocyclic.

[0425] In some embodiments, the C 3 -C 10 carbocyclic group is

[0426] . In some embodiments, the C 3 -C 10 carbocyclic group is .

[0427] In some embodiments, F 1 , F 2 or F 3 is each independently an optionally substituted C 2 -C 6 heterocyclic group. In some embodiments, the C 2– C 9 heterocyclic group is monocyclic. In some embodiments, the C 2– C 9 heterocyclic group is polycyclic. In some embodiments, the C 2– C 9 heterocyclic group is bicyclic. In some embodiments, the C 2– C 9 heterocyclic group is bridged. In some embodiments, the C 2– C 9The heterocyclic group is fused. In some embodiments, C 2– C 9 The heterocyclic group is a spirocyclic ring.

[0428] In some embodiments, C 2 -C 6 Heterocyclic group is

[0429] . In some embodiments, C 2 -C 6 Heterocyclic group is .

[0430] In some embodiments, F 1 、F 2 or F 3 Each independently is an optionally substituted C 6 -C 10 In some embodiments, F 1 、F 2 or F 3 Each independently is an optionally substituted C 2 -C 9 Heteroaryl.

[0431] In some embodiments, C 3 and C 4 Each independently is .

[0432] In some embodiments, C 3 Yes .

[0433] In some embodiments, the linker has the following structure:

[0434]

[0435]

[0436]

[0437]

[0438]

[0439]

[0440] .

[0441] ​​​​​​​​​​​In some embodiments, the compound is any one of Compounds 1-169 in Table 1, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is any one of Compounds 170-297 in Table 1, or a pharmaceutically acceptable salt thereof.

[0442] In one aspect, the compound is any one of Compounds 1-297 in Table 1, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is any one of Compounds 1-169 in Table 1, or a pharmaceutically acceptable salt thereof.

[0443]

[0444]

[0445]

[0446]

[0447]

[0448]

[0449]

[0450]

[0451]

[0452]

[0453]

[0454]

[0455]

[0456]

[0457]

[0458]

[0459]

[0460]

[0461]

[0462]

[0463]

[0464]

[0465]

[0466]

[0467]

[0468]

[0469]

[0470]

[0471]

[0472]

[0473]

[0474]

[0475]

[0476]

[0477]

[0478]

[0479]

[0480]

[0481]

[0482]

[0483]

[0484]

[0485]

[0486]

[0487]

[0488]

[0489]

[0490]

[0491]

[0492]

[0493]

[0494]

[0495]

[0496]

[0497]

[0498]

[0499]

[0500]

[0501]

[0502]

[0503]

[0504]

[0505]

[0506]

[0507]

[0508]

[0509]

[0510]

[0511]

[0512]

[0513]

[0514]

[0515]

[0516]

[0517] In one aspect, the present invention features a pharmaceutical composition comprising any of the foregoing compounds and a pharmaceutically acceptable excipient.

[0518] In another aspect, the present invention features a method of reducing the activity of the BAF complex in a cell, the method comprising contacting the cell with an effective amount of any of the foregoing compounds or a pharmaceutical composition thereof.

[0519] In some embodiments, the cell is a cancer cell.

[0520] In another aspect, the present invention features a method of treating a subject in need thereof for a BAF complex-related disorder, the method comprising administering to the subject an effective amount of any one of the foregoing compounds or a pharmaceutical composition thereof.

[0521] In some embodiments, the BAF complex-related disorder is cancer.

[0522] In another aspect, the present invention features a method of inhibiting BRM, the method comprising contacting a cell with an effective amount of any one of the foregoing compounds or a pharmaceutical composition thereof. In some embodiments, the cell is a cancer cell.

[0523] In another aspect, the present invention features a method of inhibiting BRG1, the method comprising contacting a cell with an effective amount of any one of the foregoing compounds or a pharmaceutical composition thereof. In some embodiments, the cell is a cancer cell.

[0524] In another aspect, the present invention features a method of inhibiting BRM and BRG1, the method comprising contacting a cell with an effective amount of any one of the foregoing compounds or a pharmaceutical composition thereof. In some embodiments, the cell is a cancer cell.

[0525] In another aspect, the present invention features a method of treating a subject in need thereof for a BRG1 loss-of-function mutation-related disorder, the method comprising administering to the subject an effective amount of any one of the foregoing compounds or a pharmaceutical composition thereof.

[0526] In some embodiments, the BRG1 loss-of-function mutation-related disorder is cancer. In other embodiments, the subject is determined to have a BRG1 loss-of-function disorder, such as being determined to have a BRG1 loss-of-function cancer (e.g., the cancer has been determined to include cancer cells with BRG1 loss-of-function).

[0527] In another aspect, the invention features a method of inducing apoptosis in a cell, the method comprising contacting the cell with an effective amount of any of the foregoing compounds (e.g., a BRM / BRG1 dual inhibitor compound or a BRM selective compound) or a pharmaceutical composition thereof.

[0528] In some embodiments, the cell is a cancer cell.

[0529] In another 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 foregoing compounds or a pharmaceutical composition thereof.

[0530] 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 origin, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, esophagogastric cancer, pancreatic cancer, hepatobiliary cancer, soft tissue sarcoma, ovarian cancer, head and neck cancer, renal cell cancer, bone cancer, non-Hodgkin lymphoma, small cell lung cancer, prostate cancer, embryonal tumor, germ cell tumor, cervical cancer, thyroid cancer, salivary gland cancer, gastroenteropancreatic neuroendocrine tumor, uterine sarcoma, gastrointestinal stromal tumor, CNS cancer, thymic tumor, adrenocortical carcinoma, appendiceal cancer, small intestine cancer or penile cancer.

[0531] 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 origin, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer or penile cancer.

[0532] 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 inhibitor, PD1 inhibitor, interferon therapy, BRAF inhibitor, MEK inhibitor, radiotherapy, temozolomide, irinotecan, CAR-T therapy, trastuzumab, pertuzumab, tamoxifen, capecitabine, docetaxel, platinum agents such as carboplatin, taxanes such as paclitaxel and docetaxel, ALK inhibitor, MET inhibitor, pemetrexed, albumin-bound paclitaxel, doxorubicin, gemcitabine, bevacizumab, eribulin, neratinib, PARP inhibitor, blasticidin, mTOR inhibitor, topotecan, gemcitabine, VEGFR2 inhibitor, folate receptor antagonist, fosmantel, fosbretabulin or PD-L1 inhibitor).

[0533] In some embodiments of any of the above methods, the cancer has or has been determined to have one or more BRG1 mutations. In some embodiments of any of the above methods, one or more BRG1 mutations are homozygous. In some embodiments of any of the above methods, one or more BRG1 mutations are in the ATPase catalytic domain of the protein. In some embodiments of any of the above methods, one or more BRG1 mutations are a C-terminal deletion of BRG1. In some embodiments of any of the foregoing methods, the cancer does not have or has been determined not 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 not 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.

[0534] In another aspect, the present disclosure provides a method of 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 one of the foregoing compounds or a pharmaceutically acceptable salt thereof or any of the foregoing pharmaceutical compositions. In some embodiments, the disorder is a viral infection. In some embodiments, the viral infection is by a virus from the family Retroviridae such as Lentivirus (e.g., Human Immunodeficiency Virus (HIV)) and Deltaretrovirus (e.g., Human T-cell leukemia virus I (HTLV-I), Human T-cell leukemia virus II (HTLV-II)), Hepadnaviridae (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 1 (HSV-1), Herpes simplex virus 2 (HSV-2), Human Herpesvirus 6 (HHV-6), Herpesvirus K*, CMV, Varicella-zoster virus), Papillomaviridae (e.g., Human papillomavirus (HPV, HPV E1)), Parvoviridae (e.g., Parvovirus B19), Polyomaviridae (e.g., JC virus and BK virus), Paramyxoviridae (e.g., Measles virus), Togaviridae (e.g., Rubella virus). In some embodiments, the disorder is Coffin Siris, Neurofibromatosis (e.g., NF-1, NF-2 or schwannomatosis) or multiple meningiomas.

[0535] In another aspect, the present disclosure provides a method for treating a viral infection in a subject in need thereof. The method includes administering to the subject an effective amount of any one of the foregoing compounds or a pharmaceutically acceptable salt thereof or any of the foregoing pharmaceutical compositions. In some embodiments, the viral infection is an infection by a virus selected from the group consisting of Retroviridae such as Lentivirus (e.g., Human Immunodeficiency Virus (HIV)) and Deltaretrovirus (e.g., Human T-cell Leukemia Virus I (HTLV-I), Human T-cell Leukemia Virus II (HTLV-II)), Hepadnaviridae (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 1 (HSV-1), Herpes Simplex Virus 2 (HSV-2), Human Herpesvirus 6 (HHV-6), Herpesvirus K*, CMV, Varicella-Zoster Virus), Papillomaviridae (e.g., Human Papillomavirus (HPV, HPV E1)), Parvoviridae (e.g., Parvovirus B19), Polyomaviridae (e.g., JC Virus and BK Virus), Paramyxoviridae (e.g., Measles Virus) or Togaviridae (e.g., Rubella Virus).

[0536] In some embodiments of any of the foregoing aspects, the compound is a BRM-selective compound. In some embodiments, the inhibition of the level and / or activity of BRM by the BRM-selective compound is at least 10-fold greater than the inhibition of the level and / or activity of BRG1 by the compound, and / or the binding of the compound to BRM is at least 10-fold greater than the binding of the compound to BRG1. For example, in some embodiments, the IC 50 or IP 50 of the BRM-selective compound is at least 10-fold lower than the IC 50 or IP 50 for BRG1. 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 activity of the compound against BRM and BRG1 is within 10-fold (e.g., less than 5-fold, less than 2-fold)). 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 50 or IP 50 of the BRM / BRG1 dual inhibitor compound against BRM is within 10-fold of the IC 50 or IP 50 for BRG1.

[0537] In another aspect, the invention features a method of treating melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or hematological 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.

[0538] In another aspect, the invention features a method of reducing tumor growth of melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or hematological 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.

[0539] In another aspect, the invention features a method of inhibiting the metastatic progression of melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or hematological cancer in a subject, the method comprising administering an effective amount of any one of the foregoing compounds or a pharmaceutical composition thereof.

[0540] In another aspect, the invention features a method of inhibiting the metastatic colonization (e.g., metastatic colonization to the liver and / or brain) of melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or hematological cancer in a subject, the method comprising administering an effective amount of any one of the foregoing compounds or a pharmaceutical composition thereof.

[0541] 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.

[0542] In some embodiments of any of the above aspects, the melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or blood cells are in a subject.

[0543] In some embodiments of any of the above aspects, 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. In some embodiments, an 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, an 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%).

[0544] 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%) and maintains this reduction 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 longer). 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%) and maintains this reduction for at least 4 days (e.g., 5 days, 6 days, 7 days, 14 days, 28 days, or longer).

[0545] 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%).

[0546] In some embodiments, 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%) and maintains this reduction 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 longer). In some embodiments, 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%) and maintains this reduction for at least 4 days (e.g., 5 days, 6 days, 7 days, 14 days, 28 days, or longer).

[0547] In some embodiments, the subject has cancer. In some embodiments, the cancer expresses BRG1 and / or BRM proteins, 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 hematological 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 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 cancer (e.g., the cancer has spread to the liver). Metastatic cancer may include cells that exhibit migration and / or invasion of migrating cells and / or include cells that exhibit endothelial recruitment and / or angiogenesis. In other embodiments, the cancer is a cell migration cancer. In other embodiments, the cell migration cancer is a non-metastatic cell migration cancer. Metastatic cancer may be cancer that spreads via inoculation of the surface of the peritoneum, pleura, pericardium, or subarachnoid space. Alternatively, metastatic cancer may be cancer that spreads via the lymphatic system or is hematogenous spread. In some embodiments, an effective amount of an agent that reduces the level and / or activity of BRG1 and / or BRM is an amount effective to inhibit the metastatic colonization of cancer to the liver and / or brain.

[0548] 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.

[0549] In some embodiments, the method further comprises administering to the subject or contacting the cell with an anti-cancer therapy, such as a chemotherapeutic agent or cytotoxic agent, immunotherapy, surgery, radiation therapy, thermal therapy or photocoagulation. In some embodiments, the anti-cancer therapy is a chemotherapeutic agent or cytotoxic agent, such as an antimetabolite, an anti-mitotic agent, an anti-tumor antibiotic, an asparaginase, a bisphosphonate, an anti-tumor drug, an alkylating agent, a DNA repair enzyme inhibitor, a histone deacetylase inhibitor, a corticosteroid, a demethylating agent, an immunomodulator, a janus associated kinase inhibitor, a phosphoinositide 3-kinase inhibitor, a proteasome inhibitor or a tyrosine kinase inhibitor.

[0550] In some embodiments, the compounds of the present invention are used in combination with another anti-cancer 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 present 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 present invention.

[0551] In some embodiments, the anti-cancer therapy and the compounds of the present invention are administered within 28 days of each other and each in an amount effective to treat the subject together.

[0552] In some embodiments, the subject or cancer has and / or has been identified as having a loss-of-function mutation in BRG1. In some embodiments, the subject or cancer has and / or has been identified as having a loss-of-function mutation in BRM.

[0553] In some embodiments, the cancer is resistant to one or more chemotherapeutic or cytotoxic agents (e.g., the cancer has been determined to be resistant to chemotherapeutic or cytotoxic agents, such as by genetic markers, or may be resistant to chemotherapeutic or cytotoxic agents, such as a cancer that does not respond to chemotherapeutic or cytotoxic agents). In some embodiments, the cancer does not respond to one or more chemotherapeutic or cytotoxic agents. In some embodiments, the cancer is resistant or non-responsive to dacarbazine, temozolomide, cisplatin, treosulfan, fotemustine, IMCgp100, CTLA-4 inhibitors (e.g., ipilimumab), PD-1 inhibitors (e.g., nivolumab or pembrolizumab), PD-L1 inhibitors (e.g., atezolizumab, avelumab or durvalumab), mitogen-activated protein kinase (MEK) inhibitors (e.g., selumetinib, binimetinib or trametinib), and / or protein kinase C (PKC) inhibitors (e.g., sotrastaurin or IDE196).

[0554] In some embodiments, the cancer is resistant or non-responsive 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 non-responsive to a mitogen-activated protein kinase (MEK) inhibitor (e.g., selumetinib, binimetinib or trametinib) and / or a protein kinase C (PKC) inhibitor (e.g., sotrastaurin or IDE196).

[0555] Chemical terms

[0556] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0557] For any of the following chemical definitions, the number after the atomic symbol represents the total number of atoms of the element present in the particular chemical moiety. It is understood that other atoms, such as H atoms or substituents as described herein, may be present as needed to satisfy the valency of the atoms. For example, unsubstituted C 2 An alkyl group has the formula –CH 2 CH 3 . When used with the groups defined herein, reference to the number of carbon atoms includes divalent carbons in acetal and ketal groups, but does not include the carbonyl carbon in acyl, ester, carbonate or carbamate groups. Reference to the number of oxygen, nitrogen or sulfur atoms in a heteroaryl group includes only those atoms that form part of the heterocycle.

[0558] As used herein, the term "acyl" refers to H or an alkyl group attached to a parent molecular group through a carbonyl as defined herein, and is exemplified by formyl (i.e., carboxyaldehyde group), acetyl, trifluoroacetyl, propionyl, and butyryl. Exemplary unsubstituted acyl groups include those having 1 to 6, 1 to 11, or 1 to 21 carbons.

[0559] As used herein, the term "alkyl" refers to a branched or straight-chain monovalent saturated aliphatic hydrocarbon group having 1 to 20 carbon atoms (e.g., 1 to 16 carbon atoms, 1 to 10 carbon atoms, 1 to 6 carbon atoms, or 1 - 3 carbon atoms). "Alkylene" is a divalent alkyl.

[0560] As used herein, the term "alkenyl" when used alone or in combination with other groups refers to a straight-chain or branched-chain 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). "Alkenylene" is a divalent alkenyl.

[0561] As used herein, the term "alkynyl" when used alone or in combination with other groups refers to a straight-chain or branched-chain 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). "Alkynylene" is a divalent alkynyl.

[0562] As used herein, the term "amino" represents –N(R N1 ) 2 , where each R N1 is independently H, OH, NO 2 , N(R N2 ) 2 , SO 2 OR N2 , SO 2 R N2 , SOR N2 , N -protecting group, alkyl, alkoxy, aryl, arylalkyl, cycloalkyl, acyl (e.g., acetyl, trifluoroacetyl, or other acyls as described herein), where each of these listed R N1 groups may optionally be substituted; or two R N1 groups combine to form an alkylene or heteroalkylene, and where each R N2 is independently H, alkyl, or aryl. The amino groups of the present invention can be unsubstituted amino (i.e., –NH 2 ) or substituted amino (i.e., –N(R N1 ) 2 ).

[0563] As used herein, the term "aryl" refers to an aromatic mono- or polycarbocyclic group of 6 to 12 carbon atoms having at least one aromatic ring. Examples of such groups include, but are not limited to, phenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, 1,2-dihydronaphthyl, indanyl, and 1H-indenyl. "Arylene" is a divalent aryl.

[0564] As used herein, the term "arylalkyl" denotes an alkyl group substituted with an aryl group. Exemplary unsubstituted arylalkyls are those having 7 to 30 carbons (e.g., 7 to 16 or 7 to 20 carbons, 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 aryl), such as benzyl and phenethyl. In some embodiments, the alkyl and aryl groups may each be further substituted with 1, 2, 3, or 4 substituents as defined herein for the respective groups.

[0565] As used herein, the term "azido" denotes the –N 3 group.

[0566] As used herein, the term "bridged polycycloalkyl" refers to a bridged polycyclic group of 5 to 20 carbons having 1 to 3 bridges.

[0567] As used herein, the term "cyano" denotes the -CN group.

[0568] As used herein, the term "carbocyclic group" refers to a non-aromatic C 3 -C 12 monocyclic, bicyclic, or tricyclic structure in which the ring is formed of carbon atoms. Carbocyclic structures include cycloalkyl and unsaturated carbocyclic groups.

[0569] As used herein, the term "cycloalkyl" refers to a saturated, non-aromatic, and monovalent mono- or polycarbocyclic group having 3 to 10, preferably 3 to 6 carbon atoms. This term is further illustrated by such examples as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, and adamantyl. "Cycloalkylene" is a divalent cycloalkyl.

[0570] As used herein, the term "halogen" refers to a fluorine (fluoro), chlorine (chloro), bromine (bromo), or iodine (iodo) group.

[0571] 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 may be further substituted with 1, 2, 3, or 4 substituents as described herein for alkyl groups. An example of a heteroalkyl is "alkoxy", which as used herein refers to alkyl-O- (e.g., methoxy and ethoxy). Heteroalkylene is a divalent heteroalkyl. 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, the heteroalkenyl may be further substituted with 1, 2, 3, or 4 substituents as described herein for alkenyl groups. An example of a heteroalkenyl is "alkenoxy", which as used herein refers to alkenyl-O-. Heteroalkenylene is a divalent heteroalkenyl. 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 may be further substituted with 1, 2, 3, or 4 substituents as described herein for alkynyl groups. An example of a heteroalkynyl is "alkynyloxy", which as used herein refers to alkynyl-O-. Heteroalkynylene is a divalent heteroalkynyl.

[0572] As used herein, the term "heteroallyl" represents the structure , where X is O or NR, where R is H or optionally substituted alkyl.

[0573] As used herein, the term "heteroaryl" refers to an aromatic mono- or polycyclic group of 5 to 12 atoms having at least one aromatic ring, said aromatic ring containing 1, 2, or 3 ring atoms selected from nitrogen, oxygen, and sulfur, where the remaining ring atoms are carbon. One or two ring carbon atoms of the heteroaryl may be replaced by a carbonyl group. Examples of heteroaryl are pyridyl, pyrazolyl, benzoxazolyl, benzimidazolyl, benzothiazolyl, imidazolyl, oxazolyl, and thiazolyl. "Heteroarylene" is a divalent heteroaryl.

[0574] As used herein, the term "heteroarylalkyl" denotes an alkyl group substituted with a heteroaryl. Exemplary unsubstituted heteroarylalkyls are 7 to 30 carbons (e.g., 7 to 16 carbons or 7 to 20 carbons, 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 heteroaryl). In some embodiments, each of the alkyl and heteroaryl may be further substituted with 1, 2, 3, or 4 substituents as defined herein for the respective groups.

[0575] As used herein, the term "heterocyclic group" refers to a monocyclic or polycyclic group having from 3 to 12 atoms in at least one ring, said ring containing 1, 2, 3 or 4 ring atoms selected from N, O or S, and having no aromatic ring containing any N, O or S atoms. Examples of heterocyclic groups include, but are not limited to, morpholino, thiomorpholino, furyl, piperazinyl, piperidinyl, pyranyl, pyrrolidinyl, tetrahydropyranyl, tetrahydrofuryl, and 1,3-dioxolanyl. "Heterocyclylene" is a divalent heterocyclic group.

[0576] As used herein, the term "heterocyclic group alkyl" means an alkyl group substituted by a heterocyclic group. Exemplary unsubstituted heterocyclic group alkyls are from 7 to 30 carbons (e.g., from 7 to 16 or from 7 to 20 carbons, 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 heterocyclic group). In some embodiments, the alkyl and heterocyclic groups may each be further substituted with 1, 2, 3 or 4 substituents as defined herein for the respective groups.

[0577] As used herein, the term "hydroxyalkyl" means an alkyl group substituted by an -OH group.

[0578] As used herein, the term "hydroxyl" means an -OH group.

[0579] As used herein, the term " N -protecting group" means those groups intended to protect an amino group from undesired reactions during synthesis. 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, t-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-(biphenylcarbonyl)-1-methylethoxycarbonyl, α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, diphenylmethyloxycarbonyl, t-butoxycarbonyl, diisopropylmethoxycarbonyl, isopropoxycarbonyl, ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl, cyclopentyloxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl and phenylthiocarbonyl; arylalkyls such as benzyl, triphenylmethyl and benzyloxymethyl; and silyls such as trimethylsilyl. Preferred N - protecting groups are alloc, formyl, acetyl, benzoyl, pivaloyl, t-butylacetyl, alanyl, benzenesulfonyl, benzyl, t-butoxycarbonyl (Boc) and benzyloxycarbonyl (Cbz).

[0580] As used herein, the term "nitro" means the –NO 2 group.

[0581] As used herein, the term "oxo" means the =O group.

[0582] As used herein, the term "mercapto" means the -SH group.

[0583] Alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclic group (e.g., cycloalkyl), aryl, heteroaryl, and heterocyclic group may be substituted or unsubstituted. Unless otherwise specified, when substituted, there will generally be 1 to 4 substituents. Substituents include, for example: alkyl (e.g., unsubstituted and substituted, where the substituents include any of the groups described herein, such as aryl, halo, hydroxy), aryl (e.g., substituted and unsubstituted phenyl), carbocyclic group (e.g., substituted and unsubstituted cycloalkyl), halo (e.g., fluoro), hydroxy, heteroalkyl (e.g., substituted and unsubstituted methoxy, ethoxy, or thioalkoxy), heteroaryl, heterocyclic group, amino (e.g., NH 2 or mono- or dialkylamino), azido, cyano, nitro, oxo, or mercapto. Aryl, carbocyclic group (e.g., cycloalkyl), heteroaryl, and heterocyclic group may also be substituted by alkyl (unsubstituted and substituted, such as arylalkyl (e.g., substituted and unsubstituted benzyl)).

[0584] The compounds of the present invention may have one or more asymmetric carbon atoms and may exist in the form of optically pure enantiomers, mixtures of enantiomers such as racemates, optically pure diastereoisomers, mixtures of diastereoisomers, diastereomeric racemates or mixtures of diastereomeric racemates. The optically active forms can be obtained, for example, by resolution of racemates, by asymmetric synthesis or by asymmetric chromatography (chromatography using chiral adsorbents or eluents). That is, certain of the disclosed compounds can exist in various stereoisomeric forms. Stereoisomers are compounds that differ only in their spatial arrangement. Enantiomers are pairs of stereoisomers whose mirror images are non-superimposable, most commonly because they contain an asymmetrically substituted carbon atom that acts as a chiral center. "Enantiomer" refers to one of a pair of molecules that are mirror images of each other and non-superimposable. Diastereoisomers are stereoisomers that are not related as mirror images, most commonly because 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 a compound can be prepared, for example, by separating the enantiomers from a racemate using one or more well-known techniques and methods such as chiral chromatography and separation methods based thereon. Those skilled in the art can readily determine the appropriate techniques and / or methods for separating the enantiomers of the compounds described herein from a racemic mixture. "Racemate" or "racemic mixture" refers to a compound containing two enantiomers, where such a mixture does not exhibit optical activity; that is, they do not rotate the plane of polarized light. "Geometric isomers" are isomers that differ in the orientation of the substituent atoms with respect to a carbon-carbon double bond, a cycloalkyl ring or a bridged bicyclic system. The atoms (other than H) on each side of a carbon-carbon double bond can be in the E (substituents on opposite sides of the carbon-carbon double bond) or Z (substituents on the same side) configuration. "R", "S", "S*", "R*", "E", "Z", "cis" and "trans" denote the configuration relative to the core molecule. Certain of the disclosed compounds can exist in the form of atropisomers. Atropisomers are stereoisomers that arise due to hindered rotation about a single bond, where the steric strain barrier to rotation is high enough to permit the separation of conformational isomers. The compounds of the present invention can be prepared as individual isomers by isomer-specific synthesis or resolved from mixtures of isomers. Conventional resolution techniques include forming salts of the free bases of each isomer of an isomer pair using an optically active acid (followed by fractional crystallization and regeneration of the free base), forming salts of the acid forms of each isomer of an isomer 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 isomer pair using an optically pure acid, amine or alcohol (followed by chromatographic separation and removal of the chiral auxiliary), or resolving mixtures of isomers of the starting material or final product using various well-known chromatographic methods.When naming or depicting the stereochemistry of the disclosed compounds by structure, the named or depicted stereoisomers are at least 60 wt%, 70 wt%, 80 wt%, 90 wt%, 99 wt% or 99.9 wt% relative to other stereoisomers. When naming or depicting a single enantiomer by structure, the depicted or named enantiomer is at least 60 wt%, 70 wt%, 80 wt%, 90 wt%, 99 wt% or 99.9 wt% optically pure. When naming or depicting a single diastereomer by structure, the depicted or named diastereomer is at least 60 wt%, 70 wt%, 80 wt%, 90 wt%, 99 wt% or 99.9 wt% pure by weight. The percentage of optical purity is the ratio of the weight of the enantiomer to the weight of the enantiomer plus the weight of its optical isomers. The diastereomeric purity by weight is the ratio of the weight of one diastereomer to the weight of all diastereomers. When naming or depicting the stereochemistry of the disclosed compounds by structure, the named or depicted stereoisomers are at least 60%, 70%, 80%, 90%, 99% or 99.9% pure by mole fraction relative to other stereoisomers. When naming or depicting a single enantiomer by structure, the depicted or named enantiomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% pure by mole fraction. When naming or depicting a single diastereomer by structure, the depicted or named diastereomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% pure by mole fraction. The percentage of purity by mole fraction is the ratio of the moles of the enantiomer to the moles of the enantiomer plus the moles of its optical isomers. Similarly, the percentage of purity by mole fraction is the ratio of the moles of the diastereomer to the moles of the diastereomer plus the moles of its isomers. When naming or depicting the disclosed compounds by structure without indicating stereochemistry and the compound has at least one chiral center, it is understood that the name or structure encompasses any enantiomer of the compound without the corresponding optical isomer, the racemic mixture of the compound, or a mixture enriched in one enantiomer relative to its corresponding optical isomer. When the 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 diastereomers without other diastereomers, many diastereomers without other diastereomer pairs, mixtures of diastereomers, mixtures of diastereomer pairs, mixtures of diastereomers in which one diastereomer is enriched relative to one or more other diastereomers, or mixtures of diastereomers in which one or more diastereomers are enriched relative to other diastereomers. The present invention includes all such forms.

[0585] The compounds of the present disclosure also include all isotopes of atoms present in the intermediates or end compounds. "Isotope" refers to atoms having the same atomic number but different mass numbers, which is caused by different numbers of neutrons in the atomic nucleus. For example, isotopes of hydrogen include tritium and deuterium.

[0586] Unless otherwise indicated, the structures described 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 the 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. Isotopically labeled compounds (e.g., those labeled with 3 H and 14 C) can be used in compound or substrate tissue distribution assays. Tritiated (i.e., 3 H) and carbon-14 (i.e., 14 C) isotopes are useful because of their ease of preparation and detectability. Additionally, substitution with heavier isotopes such as deuterium (i.e., 2 H) can provide certain therapeutic advantages due to greater metabolic stability (e.g., extended in vivo half-life or reduced dosage requirements). In some embodiments, one or more hydrogen atoms are replaced with 2 H or 3 H, or one or more carbon atoms are replaced with carbon enriched in 13 C or 14 C. Positron emitting isotopes such as 15 O, 13 N, 11 C, and 18 F can be used in positron emission tomography (PET) studies to examine substrate receptor occupancy. The preparation of isotopically labeled compounds is known to those of skill in the art. For example, isotopically labeled compounds can generally be prepared by following procedures similar to those disclosed herein for the compounds of the invention, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

[0587] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Methods and materials are described herein for use in the present disclosure; other suitable methods and materials known in the art may also be used. The materials, methods, and examples are illustrative only and not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.

[0588] Definition

[0589] In this application, unless the context clearly dictates otherwise, (i) the term "a" or "an" can be understood to mean "at least one"; (ii) the term "or" can be understood to mean "and / or"; and (iii) the terms "comprising" and "including" can be understood to cover the recited components or steps, whether they are presented by themselves or in combination with one or more other components or steps.

[0590] As used herein, the terms "about" and "approximately" refer to values within 10% above or below the recited value. For example, the term "about 5 nM" means a range of 4.5 to 5.5 nM.

[0591] As used herein, the term "administer" refers to the administration of a composition (e.g., a compound or a formulation comprising a compound as described herein) to a subject or system. Administration can be to an animal subject (e.g., a human) by any suitable route. For example, in some embodiments, administration can be bronchial (including by bronchial instillation), buccal, enteral, intradermal, intraarterial, intracutaneous, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intratumoral, intravenous, intraventricular, mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (including by tracheal instillation), transdermal, vaginal, and vitreous.

[0592] As used herein, the term "BAF complex" refers to the BRG1 or HRBM-associated factor complex in human cells.

[0593] As used herein, the term "BAF complex-related disorder" refers to a disorder caused by or affected by the level and / or activity of the BAF complex.

[0594] As used herein, the term "BRG1 loss-of-function mutation" refers to a mutation in BRG1 that results in a decrease in protein activity (e.g., a decrease in BRG1 activity of at least 1%, such as 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.

[0595] As used herein, the term "BRG1 loss-of-function disorder" refers to a disorder that exhibits a decrease in BRG1 activity (e.g., a decrease in BRG1 activity of at least 1%, such as a decrease in BRG1 activity of 2%, 5%, 10%, 25%, 50%, or 100%).

[0596] The term "cancer" refers to a disease caused by the proliferation of malignant neoplastic cells, such as tumors, neoplasms, carcinomas, sarcomas, leukemias, and lymphomas.

[0597] As used herein, "combination therapy" or "combined 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 particular disease or disorder. The treatment regimen defines the dose and administration period of each agent such that the effects of the individual agents overlap in the subject. In some embodiments, the delivery of two or more agents is simultaneous or concurrent, and the agents may be co-formulated. In some embodiments, two or more agents are not co-formulated and are administered in a sequential manner as part of a prescription regimen. In some embodiments, the combined administration of two or more agents or treatments results in a greater reduction in symptoms or other parameters associated with the disorder than is observed with the delivery of a single agent or treatment alone or in the absence of the agent. The effects of the two treatments may be partially additive, fully additive, or greater than additive (e.g., synergistic). The sequential or substantially simultaneous administration of each therapeutic agent can be achieved by any suitable route including, but not limited to, oral, intravenous, intramuscular, and direct absorption through mucosal tissues. The therapeutic agents can be administered by the same route or by different routes. For example, the first therapeutic agent of the combination can be administered by intravenous injection while the second therapeutic agent of the combination can be administered orally.

[0598] As used herein, "cmpd" refers to a compound.

[0599] "Determining the level of a protein or RNA" refers to directly or indirectly detecting a protein or RNA by methods known in the art. "Direct determination" refers to performing a process (e.g., assaying or testing a sample, or as defined herein the term "analyzing a sample") to obtain a physical entity or value. "Indirect determination" refers to receiving the physical entity or value from a party or source (e.g., a third-party laboratory that directly obtains the physical entity or value). Methods for measuring protein levels generally include, but are not limited to, Western blotting, immunoblotting, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunoprecipitation, immunofluorescence, surface plasmon resonance, chemiluminescence, fluorescence polarization, phosphorescence, 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 interaction with other protein ligands. Methods for measuring RNA levels are known in the art and include, but are not limited to, quantitative polymerase chain reaction (qPCR) and RNA blot analysis.

[0600] "Reducing the activity of the BAF complex" refers to reducing the level of activity associated with the BAF complex or related downstream effects. Non-limiting examples of reducing the activity of the BAF complex are Sox2 activation. The activity level of the BAF complex can be measured using any method known in the art, e.g., the method described in Kadoch et al., Cell, 2013, 153, 71-85, the method of which is incorporated herein by reference.

[0601] As used herein, the term "degrader" refers to a small molecule compound that contains a degrading moiety, wherein the compound interacts with a protein (e.g., BRG1 and / or BRM) in a manner that results in protein degradation, e.g., binding of the compound results in a reduction in the protein level in a cell or subject by at least 5%.

[0602] As used herein, the term "degrading moiety" refers to a moiety whose binding results in the 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, e.g., BRG1 and / or BRM.

[0603] "Modulating the activity of the BAF complex" means altering the activity level associated with the BAF complex (e.g., GBAF), or related downstream effects. The activity level of the BAF complex can be measured using any method known in the art, e.g., the method described in Kadoch et al., Cell 153:71-85 (2013), the method of which is incorporated herein by reference.

[0604] "Reducing the activity of BRG1 and / or BRM" refers to reducing the level of activity associated with BRG1 and / or BRM, or reducing related downstream effects. Non-limiting examples of inhibiting the activity of BRG1 and / or BRM are reducing the level of the BAF complex in a cell. The level of activity of BRG1 and / or BRM can be measured using any method known in the art. In some embodiments, the agent for reducing the activity of BRG1 and / or BRM is a small molecule BRG1 and / or BRM degrader.

[0605] "Reducing the level of BRG1 and / or BRM" refers to reducing 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.

[0606] "Level" refers to the level of a protein or the mRNA encoding the protein compared to a reference. The reference can be any useful reference as defined herein. A "reduction" or "increase in level" of a protein means a decrease or increase in the protein level compared to the reference (e.g., a decrease 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 decrease or increase of more than about 10%, about 15%, about 20%, about 50%, about 75%, about 100% or about 200% compared to the reference; a decrease or increase of less than about 0.01-fold, about 0.02-fold, about 0.1-fold, about 0.3-fold, about 0.5-fold, about 0.8-fold or less; or an increase of more than about 1.2-fold, about 1.4-fold, about 1.5-fold, about 1.8-fold, about 2.0-fold, about 3.0-fold, about 3.5-fold, about 4.5-fold, about 5.0-fold, about 10-fold, about 15-fold, about 20-fold, about 30-fold, about 40-fold, about 50-fold, about 100-fold, about 1000-fold or more). The level of a protein can be expressed as mass / volume (e.g., g / dL, mg / mL, μg / mL, ng / mL) or as a percentage relative to the total protein or mRAN in the sample.

[0607] As used herein, the term "inhibiting BRM and / or BRG1" refers to blocking or reducing the level or activity of the ATPase catalytic binding domain or the bromodomain of the protein. BRM and / or BRG1 inhibition can be determined using methods known in the art, such as, for example, the BRM ATPase assay, the Nano DSF assay, or the BRM luciferase cell assay.

[0608] As used herein, the term "pharmaceutical composition" refers to a composition containing a compound described herein formulated together with a pharmaceutically acceptable excipient and suitable for administration to a mammal such as a human. Generally, pharmaceutical compositions are manufactured or sold under the approval of a government regulatory agency as part of a treatment regimen for treating diseases in mammals. The pharmaceutical composition can be formulated, for example, for oral administration in unit dosage forms (such as tablets, capsules, cachets, soft gelatin capsules or syrups); for topical administration (such as in the form of creams, gels, lotions or ointments); for intravenous administration (such as in the form of a sterile solution free of particulate embolisms and in a solvent system suitable for intravenous use); or in any other pharmaceutically acceptable formulation.

[0609] "Pharmaceutically acceptable excipient" as used herein refers to any ingredient other than the compounds described herein (for example, a vehicle capable of suspending or dissolving the active compound) and having generally non-toxic and non-inflammatory properties in a patient. Excipients can include, for example: anti-adherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (pigments), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavoring agents, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, adsorbents, suspending or dispersing agents, sweetening agents, and water for hydration.

[0610] As used herein, the term "pharmaceutically acceptable salt" refers to any pharmaceutically acceptable salt of a compound, such as any compound of Formula I or II. Pharmaceutically acceptable salts of any compound described herein can include those salts that are suitable, within the scope of reasonable medical judgment, for contact with the tissues of humans and animals without undue toxicity, irritation, allergic response, and commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in: Berge et al., J. Pharmaceutical Sciences 66:1-19, 1977 and Pharmaceutical Salts: Properties, Selection, and Use, (edited by P.H. Stahl and C.G. Wermuth), Wiley-VCH, 2008. The salts can be prepared in situ during the final isolation and purification of the compounds described herein, or can be prepared separately by reacting the free base moiety with a suitable organic acid.

[0611] The compounds of the present invention may have ionizable groups in order to be able to be prepared as pharmaceutically acceptable salts. These salts may be acid addition salts involving inorganic or organic acids, or in the case of the compounds of the present invention in acidic form, the salts may be prepared from inorganic or organic bases. Generally, the compounds are prepared as pharmaceutically acceptable salts or used as pharmaceutically acceptable salts which are addition products of pharmaceutically acceptable acids or bases. Suitable pharmaceutically acceptable acids and bases and methods for preparing the appropriate salts are well known in the art. The salts may be prepared from pharmaceutically acceptable non-toxic acids and bases, including inorganic and organic acids and bases.

[0612] "Reference" means any useful 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 pre-determined negative control value, such as a "normal control" or a previous sample taken from the same subject; a sample from a normal healthy subject, such as normal cells or normal tissue; a sample from a subject not suffering from a disease (e.g., cells or tissue); a sample from a subject diagnosed with a disease but not yet treated with a compound of the present invention; a sample from a subject who has been treated with a compound of the present invention; or a sample of a purified protein or RNA (e.g., any of those described herein) at a known normal concentration. A "reference standard or level" means a value or number derived from a reference sample. A "normal control value" is a pre-determined value indicating a non-disease state, e.g., a value expected in a healthy control subject. Generally, 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 having a measured value within the normal control value for a particular biomarker is generally said to be "within normal limits" for said biomarker. A normal reference standard or level can be a value or number obtained from a normal subject who has not suffered from a disease or disorder (e.g., cancer); a subject who has been treated with a compound of the present invention. In a preferred embodiment, the reference sample, standard or level matches the sample subject sample by at least one of the following criteria: age, weight, sex, disease stage and general health. A standard curve of the level of a purified protein or RNA (e.g., any of those described herein) within the normal reference range can also be used as a reference.

[0613] As used herein, the term "subject" refers to any organism to which a composition according to the present invention can be administered, for example, for experimental, diagnostic, prophylactic and / or therapeutic purposes. Typical subjects include any animal (e.g., mammals such as mice, rats, rabbits, non-human primates and humans). A subject can be a person or animal who seeks or requires treatment, requests treatment, is receiving treatment, will receive treatment, or is under the care of a trained professional for a particular disease or disorder.

[0614] As used herein, the terms "treat", "treated", or "treating" refer to a therapeutic treatment or any measure that is intended to slow down (alleviate) an unwanted physiological condition, disorder, or disease or to obtain a beneficial or desired clinical outcome. Beneficial or desired clinical outcomes include, but are not limited to, alleviation of symptoms; reduction in the degree of a condition, disorder, or disease; stabilization (i.e., no worsening) of the state of a condition, disorder, or disease; delay or slowing in the onset of progression of a condition, disorder, or disease; improvement or remission (whether partial or total) of the state of a condition, disorder, or disease; improvement of at least one measurable physical parameter, which may not necessarily be discernible by the patient; or enhancement or improvement of a condition, disorder, or disease. Treatment includes eliciting a clinically significant response without an excessive level of side effects. Treatment also includes an extension of survival as compared to the expected survival in the absence of treatment. The compounds of the present invention can also be used, for example, to "preventively" treat or "prevent" a condition in a subject at increased risk of developing the condition.

[0615] As used herein, the terms "variant" and "derivative" are used interchangeably and refer to naturally occurring, synthetic, and semi-synthetic analogs of the compounds, peptides, proteins, or other substances described herein. Variants or derivatives of the compounds, peptides, proteins, or other substances described herein may retain or improve the biological activity of the original material.

[0616] Details of one or more embodiments of the invention are set forth in the following description. Other features, objects, and advantages of the invention will be apparent from the specification and claims. DETAILED DESCRIPTION OF THE INVENTION

[0618] The present disclosure features compounds that can be used to reduce the level and / or activity of BRG1 and / or BRM. These compounds can be used to modulate the activity of the BAF complex, for example, for treating BAF-related conditions, such as cancer. Exemplary compounds described herein include compounds having a structure according to Formula I:

[0619]

[0620] wherein

[0621] L is a linker;

[0622] B is a degrading moiety; and

[0623] A has a structure according to Formula II:

[0624]

[0625] wherein

[0626] X1 is N or CH,

[0627] X 2 and X 3 are independently N, CH or C(CH 3 );

[0628] R 1 is H, optionally substituted C 1 -C 6 acyl, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl, optionally substituted C 2 -C 9 heterocyclic group or -SO 2 R 6 ;

[0629] R 2 and R 5 are each independently H or optionally substituted C 1 -C 6 alkyl;

[0630] R 3 is H, optionally substituted C 1 -C 6 alkyl, or the bond between A and the linking group;

[0631] R 4 is H, optionally substituted C 1 -C 6 alkyl or optionally substituted C 1- C 6 heteroalkyl;

[0632] R 6 is optionally substituted C 1 -C 6 alkyl or -NR 7 R 8 ;

[0633] R 7 and R 8 are each independently optionally substituted C 1 -C 6 alkyl;

[0634] Het is a 5- or 6-membered heteroarylene group;

[0635] G 1 is optionally substituted C 6 -C 10 arylene, optionally substituted C 2 -C 9Hetocyclic group or optionally substituted C 2- C 9 Hetroaryl;

[0636] G 2 is absent, -O-, optionally substituted C 1 -C 6 Alkylene, optionally substituted C 1 -C 6 Alkenylene, optionally substituted C 1 -C 6 Hetroalkyl, optionally substituted C 2 -C 9 Heterocyclic C 1 -C 6 Alkylene, or optionally substituted C 2 -C 9 Heteroaryl C 1 -C 6 Alkylene;

[0637] G 3 is absent, optionally substituted C 6 -C 10 Arylene, optionally substituted C 6 -C 10 Cycloalkylene, optionally substituted C 2 -C 9 Hetocyclic group, or optionally substituted C 2 -C 9 Hetroaryl; and

[0638] A 1 is H or a bond between A and the linking group,

[0639] provided that Formula II contains one and only one bond between A and the linking group,

[0640] or a pharmaceutically acceptable salt thereof.

[0641] In some embodiments, R 3 is H or optionally substituted C 1 -C6 alkyl, A 1 is a bond between A and the linking group. In some embodiments, R 3 is a bond between A and the linking group, A 1 is H.

[0642] In some embodiments, the compound is any one of Compounds 1-297 in Table 1, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is any one of Compounds 1-169 in Table 1, or a pharmaceutically acceptable salt thereof.

[0643] This document describes other embodiments and exemplary methods for the synthesis of these compounds.

[0644] Medical use

[0645] The compounds described herein can be used in the methods of the present invention, and although not bound by theory, it is believed to exert its ability to regulate the level, state, and / or activity of the BAF complex, namely by reducing the level and / or activity of BRG1 and / or BRM proteins in the cells of a mammal. BAF complex-related disorders include, but are not limited to, disorders associated with loss-of-function mutations of BRG1 and / or BRM.

[0646] One aspect of the present invention relates to a method for treating a subject in need thereof for a disorder associated with a loss-of-function mutation of BRG1 and / or BRM, such as cancer (e.g., non-small cell lung cancer, colorectal cancer, bladder cancer, cancer of unknown primary origin, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, or penile cancer). In some embodiments, the compound is administered in an amount and for a time effective to produce one or more of the following (e.g., two or more, three or more, four or more): (a) reducing tumor size, (b) decreasing the rate of tumor growth, (c) increasing tumor cell death, (d) reducing tumor progression, (e) reducing the number of metastases, (f) decreasing the metastasis rate, (g) reducing tumor recurrence, (h) increasing the survival period of the subject, and (i) increasing the progression-free survival period of the subject.

[0647] Treating cancer can cause the size or volume of the tumor to decrease. For example, after treatment, the tumor size decreases by 5% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more) relative to the size before treatment. The tumor size can be measured by any reproducible measurement means. For example, the size of the tumor can be measured as the diameter of the tumor.

[0648] Treating cancer can further cause the number of tumors to decrease. For example, after treatment, the number of tumors decreases 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 the tumors visible to the naked eye or at a specified magnification (e.g., 2x, 3x, 4x, 5x, 10x, or 50x).

[0649] Treatment of cancer can reduce the number of metastatic nodules in other tissues or organs distant 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 the metastatic nodules visible to the naked eye or at a specified magnification (e.g., 2x, 10x or 50x).

[0650] Treatment of cancer can increase the average survival time of a group of subjects treated according to the present invention compared to a group of untreated subjects. For example, the average survival time is increased by more than 30 days (more than 60 days, 90 days or 120 days). The increase in the average survival time of the group can be measured by any reproducible means. The increase in the average survival time of the group can be measured, for example, by calculating the average survival length of the group after starting treatment with the compound of the present invention. The increase in the average survival time of the group can also be measured, for example, by calculating the average survival length of the group after completion of the first round of treatment with the pharmaceutically acceptable salt of the present invention.

[0651] Treatment of cancer can also reduce the mortality rate of a group of treated subjects compared to an untreated group. For example, the mortality rate is reduced by more than 2% (e.g., more than 5%, 10% or 25%). The reduction in the mortality rate of the group of treated subjects can be measured by any reproducible means, for example, by calculating the average number of disease-related deaths in the group per unit time after starting treatment with the pharmaceutically acceptable salt of the present invention. The reduction in the mortality rate of the group can also be measured, for example, by calculating the average number of disease-related deaths in the group per unit time after completion of the first round of treatment with the pharmaceutically acceptable salt of the present invention.

[0652] Exemplary cancers treatable 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, esophagogastric cancer, pancreatic cancer, hepatobiliary cancer, soft tissue sarcoma, ovarian cancer, head and neck cancer, renal cell cancer, bone cancer, non-Hodgkin lymphoma, prostate cancer, embryonal tumors, germ cell tumors, cervical cancer, thyroid cancer, salivary gland cancer, gastroenteropancreatic neuroendocrine tumors, uterine sarcoma, gastrointestinal stromal tumors, CNS cancers, thymic tumors, adrenocortical carcinoma, appendiceal cancer, small intestine cancer and penile cancer.

[0653] Combination preparations and their uses

[0654] The compounds of the present invention can be combined with one or more therapeutic agents. In particular, the therapeutic agent can be a therapeutic agent for treating or prophylactically treating any cancer described herein.

[0655] Combination therapy

[0656] The compounds of the present invention can be used alone or in combination with additional therapeutic agents (e.g., other agents for treating cancer or cancer-related symptoms) or in combination with other types of therapies to treat cancer. In combination therapy, the dose of one or more therapeutic compounds can be reduced from the standard dose when administered alone. For example, the dose can be determined empirically based on the drug combination and arrangement, or can be derived by isobolographic analysis (e.g., Black et al., Neurology 65:S3-S6, 2005). In such cases, the dose of the compound in combination should provide a therapeutic effect.

[0657] In some embodiments, the second therapeutic agent is a chemotherapeutic agent (e.g., a cytotoxic agent or other compound useful in treating cancer). These include alkylating agents, antimetabolites, folic acid analogs, pyrimidine analogs, purine analogs and related inhibitors, vinca alkaloids, epipodophyllotoxins, 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), leucovorin (LV), irinotecan, oxaliplatin, capecitabine, paclitaxel, and docetaxel. Non-limiting examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide; alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethyleneimines and methylmelamines including hexamethylmelamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolmelamine; polyacetylenes (especially bryostatin and bryostatinone); camptothecin (including the synthetic analog topotecan); bryostatin; spongistatin; CC-1065 (including its synthetic analogs adozelesin, carzelesin, and bizelesin); cryptophycins (especially cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including the synthetic analogs, KW-2189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil, chlornaphazine, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenester mustard, prednimustine, trophosphamide, and uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimustine; antibiotics such as enediyne antibiotics (e.g., calicheamicin, especially calicheamicin γII and calicheamicin ωII (see, e.g., Agnew, Chem.Intl. Ed Engl.33:183-186 (1994)); dynemicin, including dynemicin A; bisphosphonates, such as clodronate; esperamicin; and neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromophores), aclacinomycin, actinomycin, anthramycin, azaserine, bleomycin, actinomycin C, carubicin, carminomycin, cactinomycin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, Adriamycin® (doxorubicin, including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxyadriamycin), epirubicin, esorubicin, idarubicin, masepramycin, mitomycin (such as mitomycin C), mycophenolic acid, nogalamycin, olivomycin, peplomycin, porfiromycin, puromycin, triferrioxamine doxorubicin, rodorubicin, streptozotocin, streptozocin, tubercidin, bestatin, zinostatin, zorubicin; antimetabolites, such as methane aminopterin and 5-fluorouracil (5-FU); folic acid analogs, such as dimethylfolic acid, methotrexate, pteropterin, trimetrexate; purine analogs, such as fludarabine, 6-mercaptopurine, thioguanine, thiopurine; pyrimidine analogs, such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, didanosine, doxifluridine, enocitabine, floxuridine; androgens, such as calusterone, delmadinone acetate, cytrterone acetate, methyltestosterone, testolactone; antiadrenal drugs, such as aminoglutethimide, mitotane, trilostane; folic acid supplements, such as folinic acid; glucuronolactone; aldophosphamide glycoside; aminolevulinic acid; enuracil; aclarubicin; bexarotene; bisantrene; edatrexate; defofamine; colchicine amide; diaziquone; eflornithine; elisidepsin; epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidamine; maytansine alkaloids, such as maytansine and ansamitocin; mitoguazone; mitoxantrone; mopidamol; nitraerine; pentostatin; pipobroman; gacytosine; cytarabine (“Ara-C”); cyclophosphamide; thiotepa; taxanes, such as Taxol® paclitaxel (Bristol-Myers Squibb Oncology, Princeton, N.J.), ABraxane®, an albumin-engineered nanoparticle formulation of paclitaxel without Cremophor (American Pharmaceutical Partners, Schaumberg, Ill.) and Taxotere® docetaxel (Rhone-Poulenc Rorer, Antony, France); chlorambucil; Gemzar® gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum coordination complexes such as cisplatin, oxaliplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; Navelbine® vinorelbine; norantrone; teniposide; edatrexate; daunomycin; aminopterin; Xeloda; ibandronate; irinotecan (e.g., CPT-11); topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid; capecitabine; and pharmaceutically acceptable salts, acids or derivatives of any of the foregoing. Two or more chemotherapeutic agents may be used in the mixture for administration in combination with the first therapeutic agent described herein. Suitable dosing regimens for combination chemotherapy are known in the art and are described, for example, in Saltz et al. (1999) Proc ASCO 18:233a and Douillard et al. (2000) Lancet 355:1041-7.

[0658] In some embodiments, the second therapeutic agent is a therapeutic agent that is a biological agent for cancer treatment, such as a cytokine (e.g., interferon or interleukin (e.g., IL-2)). In some embodiments, the biological agent is an anti-angiogenic agent, such as an anti-VEGF agent, e.g., bevacizumab (Avastin®). In some embodiments, the biological agent is an immunoglobulin-based biological agent, e.g., a monoclonal antibody (e.g., a humanized antibody, a fully human antibody, an Fc fusion protein or a functional fragment thereof), which agonizes a target to stimulate an anti-cancer response or antagonizes an antigen important for cancer. Such agents include rituximab; daclizumab; basiliximab; palivizumab; infliximab; trastuzumab; gemtuzumab ozogamicin; alemtuzumab; iodine-131 tositumomab; efalizumab; cetuximab; bevacizumab; natalizumab; tocilizumab; panitumumab; ranibizumab; eculizumab; certolizumab pegol; golimumab; canakinumab; ustekinumab; ofatumumab; denosumab; motavizumab; raxibacumab; belimumab; ipilimumab; ado-trastuzumab emtansine; pertuzumab; ado-trastuzumab emtansine; and obinutuzumab. Antibody-drug conjugates are also included.

[0659] The second agent can be a non-pharmacological therapeutic agent. For example, the second therapeutic agent is radiotherapy, cryotherapy, hyperthermia, and / or surgical resection of tumor tissue.

[0660] The second agent can be a checkpoint inhibitor. In one embodiment, the inhibitor of the checkpoint 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 inhibitor of the checkpoint is a fusion protein, such as an Fc-receptor fusion protein. In some embodiments, the inhibitor of the checkpoint is an agent that interacts with a checkpoint protein, such as an antibody. In some embodiments, the inhibitor of the checkpoint is an agent that interacts with a ligand of a checkpoint protein, such as an antibody. In some embodiments, the inhibitor of the checkpoint is an inhibitor of CTLA-4 (e.g., an inhibitory antibody or a small molecule inhibitor) (e.g., an anti-CTLA4 antibody, such as ipilimumab or tremelimumab). In some embodiments, the inhibitor of the checkpoint is an inhibitor of PD-1 (e.g., an inhibitory antibody or a small molecule inhibitor) (e.g., nivolumab; pembrolizumab; pidilizumab). In some embodiments, the inhibitor of the checkpoint is an inhibitor of PDL1 (e.g., an inhibitory antibody or a small molecule inhibitor) (e.g., atezolizumab; durvalumab; avelumab; BMS 936559). In some embodiments, the inhibitor of the checkpoint is an inhibitor of PDL2 (e.g., an inhibitory antibody or an Fc fusion or a small molecule inhibitor) (e.g., a PDL2 / Ig fusion protein, such as AMP 224). In some embodiments, the inhibitor of the checkpoint is an inhibitor of B7-H3 (e.g., enoblituzumab), B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, a B-7 family ligand, or a combination thereof (e.g., an inhibitory antibody or a small molecule inhibitor).

[0661] In any of the 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 before or after the second therapeutic agent, 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 to 7, 1 to 14, 1 to 21, or 1 to 30 days.

[0662] Pharmaceutical composition

[0663] Preferably, the compounds of the present invention are formulated into pharmaceutical compositions and administered to mammals, preferably to 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.

[0664] The compounds of the present invention can be used in the form of the free base, in the form of salts, solvates and in the form of prodrugs. All forms are within the scope of the present invention. As will be understood by those skilled in the art, according to the methods of the present invention, the described compounds or their salts, solvates or prodrugs can be administered to a patient in a variety of forms depending on the chosen route of administration. The compounds of the present invention can be administered, for example, by oral, parenteral, buccal, sublingual, nasal, rectal, patch, pump or transdermal administration, and the pharmaceutical compositions are formulated accordingly. Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transdermal, nasal, intralung, intrathecal, rectal and topical modes of administration. Parenteral administration can be carried out by continuous infusion over a selected period of time.

[0665] The compounds of the present invention can be administered orally, for example, with an inert diluent or with an absorbable edible carrier, or they can be encapsulated in hard or soft gelatin capsules, or they can be compressed into tablets, or they can be incorporated directly with the foods of the diet. For oral therapeutic administration, the compounds of the present invention can be mixed with excipients and used in the form of ingestible tablets, buccal tablets, lozenges, capsules, elixirs, suspensions, syrups, and cachets. 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 with or without alcohol, and in oils. Under normal storage and use conditions, these preparations can contain preservatives to prevent the growth of microorganisms. Conventional procedures and ingredients for the selection and preparation of suitable formulations are described, for example, in Remington’s Pharmaceutical Sciences (2003, 20th edition) and The United States Pharmacopeia: The National Formulary (USP 24 NF19) published in 1999. Pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must be fluid to the extent that easy injection via a syringe is possible. Compositions for nasal administration can be conveniently formulated as aerosols, drops, gels, and powders. Aerosol formulations generally comprise a solution or fine suspension of the active substance in a physiologically acceptable aqueous or non-aqueous solvent and are usually present in a sealed container in sterile form, either as a single-dose or multi-dose form, which container can take the form of a cartridge or a refill for an atomizing device. Alternatively, the sealed container can be an integrated dispensing device, such as a single-dose nasal inhaler or an aerosol dispenser equipped with a metering valve intended to be discarded after use. When the dosage form includes an aerosol dispenser, it will contain a propellant, which can be a compressed gas (such as compressed air) or an organic propellant (such as a chlorofluorocarbon). The aerosol dosage form can also take the form of a pump nebulizer. Compositions suitable for buccal or sublingual administration include tablets, lozenges, and pastilles, in which the active ingredient is formulated with a carrier such as sugar, gum arabic, or tragacanth, gelatin, and glycerol. Compositions for rectal administration are conveniently in the form of suppositories containing a conventional suppository base such as cocoa butter. The compounds described herein can be administered intratumorally, for example, in the form of intratumoral injection. Intratumoral injection is direct injection into the tumor vasculature and is particularly contemplated for discrete, solid, accessible tumors. Local, regional, or systemic administration may also be appropriate. The compounds described herein can advantageously be contacted by administering, for example, injections spaced at approximately 1 cm intervals or multiple injections to the tumor.In the case of surgery, the present invention can be used preoperatively, for example, to subject inoperable tumors to resection. In appropriate cases, continuous administration can also be applied, for example, by implanting a catheter into the tumor or the tumor vasculature.

[0666] As described herein, the compounds of the present invention can be administered to animals (such as humans) alone or in combination with a pharmaceutically acceptable carrier, in a proportion determined by the solubility and chemical properties of the compound, the selected route of administration, and standard pharmaceutical practice.

[0667] Dosage

[0668] The dosage of the compounds of the present invention and / or the compositions containing the compounds of the present invention can vary depending on many factors, such as the pharmacodynamic properties of the compound; the mode of administration; the age, health status, 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 a suitable dosage based on the above factors. The compounds of the present invention can be initially administered at a suitable dosage, which can be adjusted as needed based on the clinical response. Generally, satisfactory results can be obtained when the compounds of the present invention are administered to humans at a daily dosage, for example, between 0.05 mg and 3000 mg (measured in solid form). Dosage ranges include, for example, between 10 - 1000 mg (such as, 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.

[0669] Alternatively, the weight of the patient can be used to calculate the amount of the dosage. For example, the dosage of the compound or its pharmaceutical composition administered to the patient can be between 0.1 - 100 mg / kg. Examples

[0670] Example 1. Synthesis of the compound of the present invention

[0671] Preparation of tert-butyl N-[2-[2-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]amino]ethoxy)ethoxy]ethyl]carbamate trifluoroacetate (I-1)

[0672]

[0673] Step 1: Preparation of tert-butyl N-[2-[2-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-iso indol-5-yl]amino]ethoxy)ethoxy]ethyl]carbamate (B)

[0674]

[0675] At 90 °C, to a stirred mixture of 2-(2,6-dioxopiperidin-3-yl)-5-fluoro-2,3-dihydro-1H-isoindole-1,3-dione (A, 3.00 g, 10.861 mmol, 1.00 equiv) and tert-butyl N-[2-[2-(2-aminoethoxy)ethoxy]ethyl]carbamate (4.05 g, 16.291 mmol, 1.50 equiv) in NMP (30 mL) was added DIEA (4.21 g, 32.58 mmol, 3.00 equiv). After 3 h, water (100 mL) was added to the mixture, which was then extracted with EtOAc (3 × 200 mL). The residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, aqueous ACN, 0%–50% gradient; detector, UV 254 nm. B was obtained as a yellow solid (1.18 g, 19.94%). LCMS (ESI) m / z: [M + H]+ = 505.

[0676] Step 2: Preparation of 5-([2-[2-(2-aminoethoxy)ethoxy]ethyl]amino)-2-(2,6-dioxopiperidin-3-yl)-2,3-dihydro-1H-isoindole-1,3-dione trifluoroacetate (I-1)

[0677]

[0678] At room temperature, TFA (2.00 mL) was added to a stirred mixture of B (1.18 g, 0.002 mmol, 1.00 equiv) in DCM (6.00 mL). The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated under reduced pressure. This afforded I-1 as a yellow solid (1.08 g, 89%). 1 1H NMR (400 MHz, DMSO-d6) δ 8.38 (d, 1H, formic acid), 7.57 (d, 1H), 7.24 (s, 1H), 7.02 (d, 1H), 6.91 (dd, 1H), 5.04 (dd, 1H), 3.58 (tt, 8H), 3.37 (q, 2H), 2.95 – 2.81 (m, 3H), 2.63 – 2.51 (m, 2H), 2.00 (ddq, 1H). LCMS (ESI) m / z: [M+H] + = 405.17.

[0679]

[0680]

[0681] Preparation of 4-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-4-yl]oxy]butyric acid (I-11)

[0682]

[0683] Step 1: Preparation of tert-butyl 4-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]butanoate (B) Preparation of (B)

[0684]

[0685] To a solution of 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindole-1,3-dione (A, 2.00 g, 7.293 mmol, 1.00 eq) and tert-butyl 4-bromobutyrate (1.95 g, 8.752 mmol, 1.2 eq) in DMF (10.00 mL) was added KI (0.12 g, 0.729 mmol, 0.1 eq) and KHCO 3 (1.10 g, 10.940 mmol, 1.5 eq), and the resulting solution was stirred at 60 °C for 5 h. The mixture was diluted with EtOAc (50 mL) and washed with water (50 mL × 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, eluting with a 0 - 32% aqueous ACN gradient to give B (1.5 g, 49.39%) as an off-white solid. LCMS (ESI) M / z [ M + H ]+ = 417.

[0686] Step 2: Preparation of 4-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-4-yl]oxy]butyric acid (I-11)

[0687]

[0688] To a stirred solution of B (450 mg, 1.081 mmol, 1 eq) in DCM (5 mL) was added TFA (1 mL). The resulting solution was stirred at 25 °C for 2 h and then concentrated. 360 mg (92.46%) of I-11 was obtained as a white solid. 1HNMR (400 MHz, methanol-d4) δ 7.79 (t, J = 8.4, 7.4 Hz, 1H), 7.47 (d, J = 7.8 Hz, 2H), 5.12 (dd, J = 12.6, 5.5 Hz, 1H), 4.30 (t, J = 6.2 Hz, 2H), 2.95 – 2.66 (m, 3H), 2.60 (t, J = 7.3 Hz, 2H), 2.25 – 2.18 (m, 3H). LCMS (ESI) m / z: [M+H] + = 361.10。

[0689]

[0690] Preparation of 3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)propanoic acid (I-17)

[0691]

[0692] Step 1: Preparation of tert-butyl 3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino ethoxy)ethoxy)propanoate (B)

[0693]

[0694] To a solution of 2-(2,6-dioxopiperidin-3-yl)-4-fluoro-2,3-dihydro-1H-isoindole-1,3-dione (A, 1.00 g, 3.620 mmol, 1.00 equiv) in NMP (10.00 mL) was added tert-butyl 3-[2-(2-aminoethoxy)ethoxy]propanoate (929.10 mg, 3.982 mmol, 1.10 equiv). The resulting mixture was stirred at 90 °C overnight. The mixture was cooled to room temperature and then diluted with EtOAc (30 mL). The solution was washed with water (10 mL x 5) and then with brine (20 mL). The resulting mixture was concentrated under reduced pressure and the residue was purified by silica gel column chromatography, eluting with petroleum ether / EtOAc (5:1 to 1:1) to give B (1.14 g, 64.33%) as a yellow solid. LCMS (ESI) m / z: [M+H] + =490。

[0695] Step 2: Preparation of 3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino ethoxy)ethoxy)propanoic acid (I-17)

[0696]

[0697] At room temperature, TFA (0.52 mL, 4.551 mmol, 3.00 equiv) was added dropwise to a stirred solution of B (1.14 g, 2.329 mmol, 1.00 equiv) in DCM (10.00 mL). The resulting mixture was stirred at room temperature for 2 h and then concentrated in vacuo. The residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 spherical column; mobile phase, aqueous ACN solution, 0% to 100% gradient, over 50 min; 70 mL / min; detector, UV 254 nm, to afford I-17 as a yellow solid (896 mg, 70.28%). 1 H NMR (400 MHz, DMSO-d6) δ 12.15 (s, 1H), 11.09 (s,1H), 7.63 – 7.55 (m, 1H), 7.15 (d, 1H), 7.05 (d, 1H), 6.61 (t, 1H), 5.06 (dd,1H), 3.65 – 3.44 (m, 8H), 2.87 (d, 1H), 2.59 (d, 2H), 2.43 (t, 2H), 2.04 (m,1H);LCMS (ESI) m / z: [M+H] + = 434.15。

[0698]

[0699] Preparation of 2-[(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]acetamido) methyl]cyclopropane-1-carboxylic acid (I-23)

[0700]

[0701] Step 1: tert-butyl 2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]acetate (B) Step 2: Preparation of [[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]acetic acid (I- Preparation

[0702]

[0703] At room temperature, K2CO3 (8.32 g, 60.168 mmol, 3 eq) was added dropwise to a stirred solution of 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindole-1,3-dione (A, 5.50 g, 20.056 mmol, 1.00 eq) and tert-butyl 2-bromoacetate (3.91 g, 20.056 mmol, 1.00 eq) in DMF (15.00 mL). The resulting mixture was washed with 3 × 200 mL EtOAc. The residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, aqueous ACN solution, 10% to 50% gradient over 10 min; detector, UV 254 nm. B was obtained as an off-white solid (3.2 g, 45.19%). LCMS (ESI) m / z: [M+H] + = 389.

[0704] 100) Step 3: Preparation of methyl 2-[(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]eth

[0705]

[0706] B (3.20 g, 8.239 mmol, 1.00 eq) and a solution of anhydrous HCl in 1,4-dioxane (15.00 mL, 493.678 mmol, 59.92 eq) were stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. I-100 was obtained as a yellow solid (1.12 g). LCMS (ESI) m / z: [M+H] + = 289.

[0707] amido)methyl]cyclopropane-1-carboxylate (C) Step 4: Preparation of 2-[(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]eth

[0708]

[0709] At room temperature, HATU (1150.07 mg, 3.025 mmol, 1.50 equiv) was added dropwise over 2 h to a stirred solution of I-100 (670.00 mg, 2.016 mmol, 1.00 equiv) and methyl 2-(aminomethyl)cyclopropane-1-carboxylate (260.44 mg, 2.016 mmol, 1.00 equiv) in DMF (15.00 mL), followed by the addition of DIEA (781.84 mg, 6.049 mmol, 3.00 equiv). The resulting mixture was concentrated under reduced pressure and the residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, aqueous ACN solution, 10% to 50% gradient over 10 min; detector, UV 254 nm. C (778.6 mg, 78.37%) was obtained as a yellow oil. LCMS (ESI) m / z: [M+H] + = 444.

[0710] amido)methyl]cyclopropane-1-carboxylic acid (I-23) Step 1: Preparation of tert-butyl N-(2-[[2-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy

[0711]

[0712] A mixture of C (763.90 mg, 1.00 equiv) in 1,4-dioxane (5.00 mL) and a mixture of anhydrous HCl in 1,4-dioxane (4 M, 5.00 mL) were stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure and the residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, aqueous ACN solution, 10% to 50% gradient over 10 min; detector, UV 254 nm. Compound I-23 (338.2 mg) was obtained as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 12.17 (s, 1H), 11.12 (s, 1H), 8.33 (t,1H), 7.88 (d, 1H), 7.46 (d, 1H), 7.39 (dd, 1H), 5.13 (dd, 1H), 4.74 (s, 2H),2.96 – 2.83 (m, 1H), 2.65 – 2.52 (m, 1H), 2.08 (s, 4H), 1.66 (td, 1H), 1.48(h, 1H), 1.02 (td, 1H), 0.85 (dt, 1H).LCMS (ESI) m / z: [M+H] + = 430.05.

[0713] Preparation of 2-[(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]acetamido)methyl]cyclopropane-1-carboxylic acid (I-24)

[0714]

[0715] I-24 (423 mg, 35.74%) was prepared as a white solid in a similar manner to that described for the preparation of Intermediate I-23. 1 H NMR (300 MHz, DMSO-d6) δ 12.16 (s, 1H), 11.12 (s, 1H), 8.10 (s,1H), 7.82 (t, 1H), 7.51 (d, 1H), 7.41 (d, 1H), 5.17 – 5.07 (m, 1H), 4.80 (s,2H), 2.60 (d, 2H), 2.08 (s, 3H), 1.70 – 1.60 (m, 1H), 1.47 (d, 1H), 1.03 (d,1H), 0.84 (d, 1H); LCMS (ESI) m / z: [M+H] + = 430.12

[0716] Preparation of N-[2-[(2-aminoethyl)(methyl)amino]ethyl]-2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]acetamide (I-25)

[0717]

[0718] acetamido)ethyl](methyl)amino]ethyl)carbamate (A) Step 2: Preparation of N-[2-[(2-aminoethyl)(methyl)amino]ethyl]-2-[[2-(2,6-dioxopiperidin-3-

[0719]

[0720] At 25 °C, HATU (1.65 g, 4.350 mmol, 1.50 eq) was added to a stirred solution of I-100 (963.50 mg, 2.900 mmol, 1.00 eq) and tert-butyl N-[2-[(2-aminoethyl)(methyl)amino]ethyl]carbamate (945.24 mg, 4.350 mmol, 1.50 eq) in DCM over 2 h, followed by the dropwise addition of DIEA (1.12 g, 8.699 mmol, 3.00 eq). The residue was purified by silica gel column chromatography, eluting with petroleum ether / EtOAc (1:1) to afford A (898.6 mg, 58.30%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 532。

[0721] yl)-1,3-dioxoisoindol-5-yl]oxy]acetamide formate (I-25) Step 1: Preparation of tert-butyl N-(4-hydroxybutyl)carbamate (B)

[0722]

[0723] TFA (1.00 mL, 13.463 mmol, 23.16 eq) was added to a stirred mixture of B (309.00 mg, 0.581 mmol, 1.00 eq) in DCM (4.00 mL, 62.920 mmol, 108.24 eq). The solution was stirred at room temperature for 2 h under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure and then purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, aqueous ACN, 10% to 50% gradient over 35 min; detector, UV 254 nm. I-25 (268 mg, 93.71%) was obtained as a pale yellow solid. 1 H NMR (300 MHz, methanol-d4) δ 7.88(d, 1H), 7.53 (d, 1H), 7.46 (dd, 1H), 5.14 (dd, 1H), 4.82 (s, 2H), 3.75 (t,2H), 3.60 (d, 2H), 3.46 (q, 4H), 3.03 (s, 3H), 2.90-2.64 (m, 3H), 2.22 – 2.07(m, 1H).LCMS (ESI) m / z: [M+H] + = 432.20。

[0724] Preparation of N-[2-[(2-aminoethyl)(methyl)amino]ethyl]-2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-4-yl]oxy]acetamide (I-26)

[0725]

[0726] I-26 (200 mg, 41.07%) as a yellow solid was prepared in a similar manner as described for the preparation of I-25. 1 H NMR (400 MHz, DMSO-d6) δ 11.13 (s, 1H), 11.02 (s, 1H), 8.50 – 8.42(m, 4H), 7.85 – 7.77 (m, 1H), 7.49 (dd, 2H), 5.12 (dd, 1H), 4.90 (s, 2H),3.65 – 3.43 (m, 7H), 2.90 (m, 1H), 2.84 (s, 3H), 2.65 – 2.51 (m, 2H), 2.04(dtt, 1H), 1.33 – 1.19 (m, 2H);LCMS (ESI) m / z: [M+H] + = 432.18.

[0727] Preparation of 4-[[3-(4-aminobutanesulfonyl)propyl]amino]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione formate (I-27)

[0728]

[0729] Step 2: Preparation of tert-butyl N-[4-[(4-methylbenzenesulfonyl)oxy]butyl]carbamate (C)

[0730]

[0731] Di-tert-butyl dicarbonate (52.89 g, 242.321 mmol, 1.5 equiv) was added to a stirred solution of 4-aminobutan-1-ol (A, 14.40 g, 161.547 mmol, 1.00 equiv) in THF (160.00 mL), and the mixture was stirred at room temperature for 1 h. The solution was concentrated to dryness, and the oily residue was purified by flash column chromatography (40 - 60% EtOAc - hexane) to afford a colorless oil which solidified to a white solid on standing (B, 30.58 g, 100.02%); LCMS (ESI) m / z: [M+H] + = 190.

[0732] Step 3: Preparation of tert-butyl N-[4-(acetylthio)butyl]carbamate (D)

[0733]

[0734] At 0 °C, DMAP (2.96 g, 24.237 mmol, 0.15 eq), TEA (40.88 g, 403.952 mmol, 2.5 eq) and p-toluenesulfonyl chloride (46.21 g, 242.371 mmol, 1.5 eq) were added to a stirred solution of B (30.58 g, 161.581 mmol, 1.00 eq) in DCM (400.00 mL). The resulting mixture was stirred at 0 °C for 2 h, then warmed to room temperature and stirred for an additional 5 h. The mixture was concentrated and the residue was purified by silica gel column chromatography, eluting with petroleum ether / THF (1:1) to give C (45.6 g, 82.17%) as a pale yellow oil; LCMS (ESI) m / z: [M+H] + = 344.

[0735] Step 4: Preparation of benzyl N-[3-([4-[(tert-butoxycarbonyl)amino]butyl]thio)propyl]carbamate (E)

[0736]

[0737] To a stirred solution of C (45.60 g, 132.77 mmol, 1.00 eq) in ACN (300.00 mL) was added Schiff reagent (15.16 g, 199.161 mmol, 1.5 eq) and K 2 CO 3 (55.05 g, 398.323 mmol, 3 eq). The resulting mixture was stirred at room temperature for 12 h, treated with 1 M HCl, then extracted into DCM and concentrated. The residue was purified by silica gel column chromatography, eluting with petroleum ether / THF (1:1), to give D (28.7 g, 87.39%) as a pale yellow oil; LCMS (ESI) m / z: [M+H] + = 248.

[0738] Step 5: Preparation of benzyl N-(3-[4-[(tert-butoxycarbonyl)amino]butanesulfonyl]propyl)carbamate (F) ​

[0739]

[0740] To a solution of D (3.60 g, 14.554 mmol, 1.00 equiv) in MeOH (90.00 mL) was added benzyl N-(3-bromopropyl)carbamate (4.36 g, 16.010 mmol, 1.1 equiv) and NaOMe (3.15 g, 58.217 mmol, 4 equiv). The resulting solution was stirred at room temperature for 3 h, and the reaction was quenched with water at room temperature. The resulting mixture was extracted with EtOAc (3 × 100 mL). The combined organic layers were washed with brine (3 × 100 mL) and dried over anhydrous Na 2 SO 4 and filtered. 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, NH 4 HCO 3 aqueous solution, 0% - 100% gradient, over 30 min; detector, UV 254 nm. E was obtained as a pale yellow oil (4.122 g, 71.42%); LCMS (ESI) m / z: [M+H] + = 397.

[0741] ​ ​

[0742]

[0743] To a solution of E (4.13 g, 10.415 mmol, 1.00 equiv) in MeOH (60.00 mL) was added Oxone® (3.50 g, 20.840 mmol, 2 equiv). The resulting solution was stirred at room temperature for 12 h. Subsequently, water treatment and DCM extraction were performed, and the concentrated residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, NH 4 HCO 3 aqueous solution, 0% - 100% gradient, over 30 min; detector, UV 254 nm. F was obtained as a white solid (2 g, 44.81%); LCMS (ESI) m / z: [M+H] + = 429.

[0744] Step 6: Preparation of tert-butyl N-[4-(3-aminopropylsulfonyl)butyl]carbamate (G)

[0745]

[0746] To a solution of F (1.95 g, 4.550 mmol, 1.00 equiv) in ethanol (30.00 mL) was added ammonium formate (573.85 mg, 9.101 mmol, 2 equiv) and Pd(OH) on carbon 2 (6.962 mmol, 1.53 equiv). The resulting suspension was stirred at 60 °C under 1 atm of hydrogen for 12 h. The resulting mixture was filtered and the cake was washed with MeOH (3 × 30 mL). The filtrate was concentrated under reduced pressure to give G (1.12 g, crude product) as a black solid; LCMS (ESI) m / z: [M+H] + = 295.

[0747] Step 7: Preparation of tert-butyl N-[4-(3-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]amino] propylsulfonyl)butyl]carbamate (H)

[0748]

[0749] To a solution of G (1.12 g, 3.804 mmol, 1.00 equiv) in NMP (30.00 mL) was added 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (1.05 g, 3.804 mmol, 1 equiv) and DIEA (1.48 g, 11.413 mmol, 3 equiv). The resulting solution was stirred at 90 °C under a nitrogen atmosphere for 3 h. The resulting mixture was diluted with water (30 mL) and extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with brine (3 × 30 mL) and dried over anhydrous Na 2 SO 4 2. After filtration, the filtrate was concentrated under reduced pressure and the residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN / 0.1% aqueous formic acid, 0% to 100% gradient over 45 min; detector, UV 254 nm. This gave H (970 mg, 46.31%) as a yellow solid; LCMS (ESI) m / z: [M+H] + = 551.

[0750] Step 8: Preparation of 4-[[3-(4-aminobutylsulfonyl)propyl]amino]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione formate (I-27) Step d1: Preparation of tert-butyl N-(2-[[2-(1,3-dioxoisoindol-2-yl)ethyl]sulfanyl]ethyl)carbamate

[0751]

[0752] To a solution of H (970.00 mg, 1.762 mmol, 1.00 equiv) in 1,4-dioxane (5 mL) was added anhydrous HCl in 1,4-dioxane (164.559 mmol, 93.41 equiv). The resulting solution was stirred at room temperature for 3 h. After concentration, the residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN / 0.1% aqueous formic acid, 0% to 100% gradient over 45 min; detector, UV 220 nm. This gave I-27 as a yellow solid (712 mg, 89.17%); 1 H NMR (300 MHz, methanol-d4) δ 8.55–8.48 (m, 1H, formic acid), 7.59 (ddd, 1H), 7.12 (ddd, 2H), 5.08 (dd, 1H), 3.56 (t,2H), 3.33 – 3.14 (m, 4H), 3.04 – 2.93 (m, 2H), 2.96 – 2.63 (m, 3H), 2.24 –2.06 (m, 3H), 2.00 – 1.75 (m, 4H).; LCMS (ESI) m / z: [M+H] + = 451.16。

[0753] Preparation of 4-[[2-(2-Aminoethylsulfonyl)ethyl]amino]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione trifluoroacetate (I-28)

[0754]

[0755] (B) Step 2: Preparation of tert-butyl N-[2-[2-(1,3-dioxoisoindol-2-yl)ethanesulfonyl]ethyl]carbamate (C)

[0756]

[0757] Under a nitrogen atmosphere at 70 °C, to a stirred mixture of tert-butyl N-(2-mercaptoethyl)carbamate (A, 5.00 g, 28.207 mmol, 1.00 equiv) and N-(2-bromoethyl)phthalimide (7.17 g, 0.028 mmol,1.00 equiv) in ACN (10.00 mL) was added K 2 CO 3(11.70 g, 0.085 mmol, 3.00 equivalents). After 5 h, the resulting mixture was extracted with EtOAc (3 × 500 mL). The combined organic layers were washed with brine (3 x 300 mL) and dried over anhydrous Na 2 SO 4 and filtered. After filtration, the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography, eluting with petroleum ether / EtOAc (5:1 to 1:1) to give B as a white solid (8.20 g, 82.96%). LCMS (ESI) m / z: [M+H] + = 351.

[0758] Step 3: Preparation of tert-butyl N-[2-(2-aminoethanesulfonyl)ethyl]carbamate (D) Step 4: Preparation of tert-butyl N-[2-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]amino]

[0759]

[0760] Under a nitrogen atmosphere, m-CPBA (12.11 g, 70.199 mmol, 3.00 equivalents) was added to a stirred mixture of B (8.20 g, 23.400 mmol, 1.00 equivalent) in DCM (100 mL) at room temperature. The reaction was quenched with saturated Na 2 S 2 O 3 aqueous solution (100 mL) at room temperature. Saturated NaHCO 3 aqueous solution (100 mL) was added to the mixture, which was then extracted with EtOAc (3 × 400 mL) and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with petroleum ether / EtOAc (5:1 to 1:1) to give C as a white solid (8.40 g, 87.30%). LCMS (ESI) m / z: [M+H] + = 383.

[0761] ethanesulfonyl)ethyl]carbamate (E)

[0762]

[0763] At 80 °C, hydrazine hydrate (0.89 g, 17.781 mmol, 2.00 equivalents) was added to a stirred mixture of C (3.40 g, 8.891 mmol, 1.00 equivalent) in EtOH (100 mL). The resulting mixture was stirred at 80 °C for 1 h under a nitrogen atmosphere. The suspension was filtered and the cake was washed with EtOH (100 mL). The filtrate was concentrated under reduced pressure to give D as a white solid (crude product, 1.88 g, 77.94%). LCMS (ESI) m / z: [M+H]+ = 253。

[0764] Step 5: Preparation of 4-[[2-(2-aminoethanesulfonyl)ethyl]amino]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione; trifluoroacetate (I-28) Step 1: Preparation of 2-[2-[(4-methylphenylsulfonyl)oxy]ethoxy]ethanol (B)

[0765]

[0766] At 90 °C and under a nitrogen atmosphere, DIEA (2.89 g, 22.352 mmol, 3.00 equivalents) was added dropwise to a mixture of D (1.88 g, 7.451 mmol, 1.00 equivalent) and 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindole-1,3-dione (2.26 g, 8.196 mmol, 1.10 equivalents) in NMP (25.00 mL). The resulting mixture was stirred for 12 h and then extracted with EtOAc (3 × 300 mL). The combined organic layers were washed with brine (3 × 100 mL) and dried over anhydrous Na 2 SO 4 2SO4. After filtration, the filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography, eluting with petroleum ether / EtOAc (1:1), to give E (1.58 g, 40.03%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 509。

[0767] Step 2: Preparation of 2-[2-(methylamino)ethoxy]ethanol (C) Step 3: Preparation of benzyl N-[2-(2-hydroxyethoxy)ethyl]-N-methylcarbamate (D)

[0768]

[0769] Under a nitrogen atmosphere, trifluoroacetaldehyde (5.0 mL) was added dropwise to a stirred mixture of E (1.54 g, 3.028 mmol, 1.00 equivalent) in DCM (20 mL) at 25 °C. After 1 h, the resulting mixture was concentrated in vacuo. I-28 (1.68 g, 122.25%) was produced as a yellow solid. 11H NMR (400 MHz, DMSO-d6) δ 11.11 (s, 1H), 7.95 (s, 4H), 7.65 (dd, 1H), 7.18 (d, 1H), 7.12 (d, 1H), 6.84 (t, 1H), 5.07 (dd, 1H), 3.80 (q, 2H), 3.63 (m, 3H), 3.51 (t, 7H), 3.29 (dt, 4H), 2.89 (ddd, 1H), 2.70 (s, 1H), 2.65 – 2.52 (m, 2H), 2.18 (t, 1H), 2.03 (ddd, 1H), 1.96 – 1.84 (m, 1H). LCMS (ESI) m / z: [M+H] + = 409.11。

[0770] Preparation of 4-(2-[2-[(2-aminoethyl)(methyl)amino]ethoxy]ethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione formate (I-29)

[0771]

[0772] Step 4: Preparation of benzyl N-methyl-N-(2-[2-[(4-methylphenylsulfonyl)oxy]ethoxy]ethyl)carbamate

[0773]

[0774] At 0 °C, diethylene glycol (A, 12.72 g, 119.863 mmol, 1.00 equivalent) was dissolved in THF (105.00 mL). NaOH (3.60 g, 89.897 mmol, 0.75 equivalent) in 45 mL of water was added dropwise to this solution. After stirring for 30 minutes, the solution was added dropwise at 0 °C to a solution of p-toluenesulfonyl chloride (11.43 g, 59.932 mmol, 0.5 equivalent) in THF (45.00 mL). After the addition was complete, the aqueous solution was treated with 10% HCl and then extracted with dichloromethane. The organic layer was washed with distilled water and dried over MgSO 4 4. The residue was purified by column chromatography (petroleum ether / THF = 1:1 v / v). B was obtained as a colorless oil (12.56 g, 40.26%); LCMS (ESI) m / z: [M+H] + = 261。

[0775] (E)

[0776]

[0777] Dissolve B (12.56 g, 48.252 mmol, 1.00 equivalent) in 1,4-dioxane (180.00 mL), and then add methylamine hydrochloride (32.58 g, 482.520 mmol, 10 equivalents). Add K 2 CO 3 (33.34 g, 241.260 mmol, 5 equivalents) portionwise, and stir the reactants at 50 °C overnight. Remove the solvent under reduced pressure to obtain C (10 g, 73.22%) as a colorless oil; LCMS (ESI) m / z: [M+H] + = 120.

[0778] Step 5: Preparation of benzyl N-[2-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]

[0779]

[0780] Add benzyl chloroformate (2.29 g, 13.427 mmol, 0.16 equivalent), K 2 CO 3 (3.83 g, 27.693 mmol, 0.33 equivalent), and water (30.00 mL) to a solution of C (10.00 g, 83.918 mmol, 1.00 equivalent) in THF (30.00 mL). Stir the resulting mixture at room temperature for 12 hours, and then extract with EtOAc (3 × 60 mL). Wash the combined organic layers with brine (3 × 60 mL) and dry over anhydrous Na 2 SO 4 . After filtration, concentrate the liquid under reduced pressure and purify by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, NH 4 HCO 3 aqueous solution, 0% to 100% gradient over 30 minutes; detector, UV 254 nm. This gives D (2.14 g, 17.51%) as a colorless oil; LCMS (ESI) m / z: [M+H] + = 254.

[0781] ethoxy)ethyl]-N-methylcarbamate (F) Step 6: Preparation of 2-(2,6-dioxopiperidin-3-yl)-4-[2-[2-(methylamino)ethoxy]ethoxy]isoindole-1,3-dione (G)

[0782]

[0783] At 0 °C, DMAP (0.15 g, 1.267 mmol, 0.15 equiv), TEA (2.14 g, 21.121 mmol, 2.5 equiv) and p-toluenesulfonyl chloride (2.42 g, 12.673 mmol, 1.5 equiv) were added to a stirred solution of D (2.14 g, 8.449 mmol, 1.00 equiv) in DCM (30.00 mL). The resulting mixture was stirred at 0 °C for 2 h and then at room temperature for an additional 5 h. The residue was purified by silica gel column chromatography, eluting with petroleum ether / THF (1:1) to afford E (3.51 g, crude product) as a colorless oil; LCMS (ESI) m / z: [M+H] + = 408.

[0784] Step 7: Preparation of tert-butyl N-(2-[[2-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy ethoxy)ethyl](methyl)amino]ethyl)carbamate (H)

[0785]

[0786] To a solution of E (3.51 g, 8.614 mmol, 1.00 equiv) in DMF (30.00 mL) was added 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione (2.36 g, 8.614 mmol, 1 equiv) and Na 2 CO 3 (1.37 g, 12.921 mmol, 1.5 equiv). The resulting mixture was stirred at 80 °C for 12 h and then concentrated. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, CH 3 CN aqueous solution, 0% to 100% gradient over 30 min; detector, UV 254 nm. This gave F (1.527 g, 34.79%) as a pale yellow solid; LCMS (ESI) m / z: [M+H] + = 510.

[0787] Step 8: Preparation of 4-(2-[2-[(2-aminoethyl)(methyl)amino]ethoxy]ethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione formate (I-29) Step 1: Preparation of tert-butyl 3-[[(4-methylphenylsulfonyl)oxy]methyl]azetidine-1-carboxylate (B)

[0788]

[0789] To a solution of F (1.52 g, 2.997 mmol, 1.00 equiv) in EtOH (30.00 mL) was added ammonium formate (377.95 mg, 5.994 mmol, 2 equiv) and Pd(OH) 2 / C (5.425 mmol, 1.81 equivalents). The resulting suspension was stirred at 60 °C under 1 atm of hydrogen for 12 h and then filtered. The filter cake was washed with MeOH (3 x 30 mL), and the filtrate was concentrated under reduced pressure. This gave G (1.062 g, 94.83%) as a pale yellow solid; LCMS (ESI) m / z [M+H] + = 376.

[0790] ​ ​

[0791]

[0792] To a solution of G (1.06 g, 2.824 mmol, 1.00 equivalent) in DMF (10.00 mL) was added tert-butyl N-(2-oxoethyl)carbamate (539.41 mg, 3.389 mmol, 1.2 equivalents) and NaBH(OAc) 3 (1.80 g, 8.471 mmol, 3 equivalents). The resulting solution was stirred at room temperature for 2 h, then treated with water, extracted with DCM, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with DCM / MeOH (10:1) to give H (386 mg, 26.36%) as a pale yellow solid; LCMS (ESI) m / z: [M+H] + = 519.

[0793] ​ ​

[0794]

[0795] To a solution of H (511.00 mg, 0.985 mmol, 1.00 equivalent) in DCM (5.00 mL) was added TFA (5.00 mL, 67.315 mmol, 68.31 equivalents). The resulting solution was stirred at room temperature for 3 h. The solution was concentrated, and the residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, CH 3 CN aqueous solution, 0% to 100% gradient over 45 min; detector, UV 220 nm. This gave I-29 (307 mg, 73.86%) as a brown oil; 11H NMR (300 MHz, DMSO-d6) δ 11.70 - 10.30 (m, 1H), 8.18 (s, 1H, formic acid), 7.83 (dd, 1H), 7.51 (dd, 2H), 5.09 (dd, 1H), 4.41 – 4.32 (m, 2H), 3.84 – 3.75 (m, 2H), 3.63 (d, 2H), 2.99 – 2.80 (m, 3H), 2.65 - 2.58 (m, 5H), 2.24 (s, 3H), 2.08 (s, 1H), 1.98 - 2.06 (m, 1H); LCMS (ESI) m / z: [M+H] + = 419。

[0796] Preparation of 4-(azetidin-3-ylmethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione trifluoroacetate (I-30)

[0797]

[0798] ​

[0799]

[0800] At 0 °C, DMAP (0.18 g, 1.498 mmol, 0.15 equiv), TEA (2.53 g, 24.968 mmol, 2.50 equiv) and p-toluenesulfonyl chloride (2.86 g, 14.981 mmol, 1.50 equiv) were added to a stirred solution of tert-butyl 3-(hydroxymethyl)azetidine-1-carboxylate (A, 1.87 g, 9.987 mmol, 1.00 equiv) in DCM (50.00 mL). The resulting mixture was stirred at 0 °C for 2 h and then at room temperature for an additional 5 h. After workup with water and concentration, the residue was purified by silica gel column chromatography, eluting with petroleum ether / THF (1:1) to give B as a colorless oil (2.65 g, 77.72%); LCMS (ESI) m / z: [M+H] + = 342。

[0801] Step 2: Preparation of tert-butyl 3-([[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]methyl)azetidine-1-carboxylate (D) Preparation of tert-butyl 3-([[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]methyl)azetidine-1-carboxylate (D)

[0802]

[0803] To a solution of C (2.30 g, 8.387 mmol, 1.00 equiv) in DMF (15.00 mL) was added tert-butyl 3-[[(4-methylphenylsulfonyl)oxy]methyl]azetidine-1-carboxylate (B, 2.86 g, 8.387 mmol, 1 equiv) and Na 2 CO 3 (1.33 g, 12.581 mmol, 1.5 equiv). The resulting mixture was stirred at 80 °C for 5 h under 1 atm of hydrogen. The reaction was quenched with water at room temperature and extracted with EtOAc (3 × 100 mL). The combined organic layers were washed with brine (3 x 100 mL) and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. This gave D (3.37 g, 90.61%) as a pale yellow solid; LCMS (ESI) m / z: [M+H] + = 444.

[0804] Step 3: Preparation of 4-(azetidin-3-ylmethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione trifluoroacetate (I-30) Preparation of 4-(azetidin-3-ylmethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione trifluoroacetate (I-30)

[0805]

[0806] To a solution of D (2.39 g, 5.389 mmol, 1.00 equiv) in DCM (10.00 mL) was added TFA (10.00 mL, 134.630 mmol, 42.34 equiv). The resulting solution was stirred at room temperature for 3 h and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN aqueous solution, 0% to 100% gradient, over 45 min; detector, UV 220 nm. This gave I-30 (1.712 g, 87.72%) as a pale yellow solid; 11H NMR (300 MHz, DMSO-d6) δ 11.80 - 10.25 (m, 1H), 9.65 - 8.35 (m, 1H), 7.87 (dd, 1H), 7.54 (dd, 2H), 5.11 (dd, 1H), 4.39 (d, 2H), 4.09 (dd, 2H), 3.96 (dd, 2H), 3.37 – 3.24 (m, 1H), 2.99 - 2.81 (m, 1H), 2.66 - 2.61 (m, 1H), 2.61 – 2.53 (m, 1H), 2.11 - 1.98 (m, 1H); LCMS (ESI) m / z: [M+H] + = 344.12。

[0807] Preparation of 5-(azetidin-3-ylmethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione formate (I-31)

[0808]

[0809] I-31 was prepared as a white solid (906.2 mg, 81.38%) in a similar manner as described for the preparation of I-30. 1 1H NMR (300 MHz, DMSO-d6) δ 8.40 (s, 0.4H, formic acid), 7.88 (d, 1H), 7.52 (d, 1H), 7.42 (dd, 1H), 5.13 (dd, 1H), 4.35 (d, 2H), 4.08 (t, 2H), 3.85 (t, 2H), 3.22 (d, 1H), 3.00 - 2.80 (m, 1H), 2.67 - 2.61 (m, 1H), 2.60 - 2.54 (m, 1H), 2.14 – 1.99 (m, 1H).; LCMS (ESI) m / z: [M+H]+ = 344.12。

[0810] 4-[2-(azetidin-3-yl)ethoxy]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione trifluoroacetate (I-32) Preparation of

[0811]

[0812] Step 1: Preparation of tert-butyl 3-(2-ethoxy-2-oxoethylidene)azetidine-1-carboxylate (B)

[0813]

[0814] Under nitrogen, tert-butyl 3-oxoazetidine-1-carboxylate (A, 2.50 g, 14.603 mmol, 1.00 equiv) and ethyl 2-(triphenyl-λ5-phosphoranyl)acetate (5.60 g, 16.063 mmol, 1.10 equiv) were added to a Schlenk flask. Anhydrous toluene (100 mL) was added, and the mixture was heated to reflux for 2 h. After cooling to room temperature, the resulting mixture was concentrated and purified by silica gel column chromatography (cyclohexane:EtOAc, 5:1). B (3 g, 85.14%) was obtained as a colorless oil. LCMS (ESI) m / z[M+H] + = 304.

[0815] Step 2: Preparation of tert-butyl 3-(2-ethoxy-2-oxoethylidene)azetidine-1-carboxylate (C)

[0816]

[0817] At room temperature and 1 atm of hydrogen, 10% Pd / C (500 mg) was added to a stirred solution of B (3.0 g, 9.8 mmol, 1.0 equiv) in EtOH (50 mL). The reaction mixture was stirred at room temperature for 12 h, and then the catalyst was removed by filtration. The filtrate was concentrated in vacuo to give C (2.6 g, 76.16%) as a yellow oil. LCMS (ESI) m / z: [M+H] + = 216.

[0818] Step 3: Preparation of tert-butyl 3-(2-hydroxyethyl)azetidine-1-carboxylate (D)

[0819]

[0820] Under a nitrogen atmosphere and at 0 °C, LiAlH4 (202.79 mg, 5.343 mmol, 1.00 equiv) was added portionwise to a stirred solution of C (1.30 g, 5.343 mmol, 1.00 equiv) in THF (15.00 mL). The reaction was quenched by the addition of EtOAc at 0 °C. The resulting mixture was filtered, and the filter cake was washed with EtOAc. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give D (1 g, 92.99%) as a colorless oil, LCMS (ESI) m / z: [M+H] + = 202.

[0821] Step 4: Preparation of tert-butyl 3-[2-[(4-methylbenzenesulfonyl)oxy]ethyl]azetidine-1-carboxylate (E) Preparation

[0822]

[0823] D (950.00 mg, 4.720 mmol, 1.00 equiv) was added to a solution of p-toluenesulfonyl chloride (1079.81 mg, 5.664 mmol, 1.20 equiv) in anhydrous DCM (15 mL) at room temperature, and then TEA (573.15 mg, 5.664 mmol, 1.20 equiv) was added dropwise. The mixture was stirred at room temperature for 2 h and then concentrated. The residue was purified by silica gel column chromatography (gradient elution 1:15 to 1:5 EtOAc / hexane) to give E (1.5 g, 90.99%) as a colorless oil, LCMS (ESI) m / z: [M+H] + = 356.

[0824] Step 5: Preparation of tert-butyl 3-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]ethyl)azetidine-1-carboxylate (F) Preparation of tert-butyl 3-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]ethyl)azetidine-1-carboxylate (F)

[0825]

[0826] A solution of E (800.00 mg, 2.251 mmol, 1.00 equiv), 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindole-1,3-dione (617.21 mg, 2.251 mmol, 1.00 equiv) and Na 2 CO 3 (357.82 mg, 3.376 mmol, 1.50 equiv) in DMF (3 mL) was stirred at 80 °C for 2 h. The resulting mixture was extracted with EtOAc, and the combined organic layers were washed with brine and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with EtOAc, to give F (500 mg, 80.12%) as a colorless oil, LCMS (ESI) m / z: [M+H] + = 458.

[0827] Step 6: Preparation of 4-[2-(azetidin-3-yl)ethoxy]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione trifluoroacetate (I-32) Preparation of 4-[2-(azetidin-3-yl)ethoxy]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione trifluoroacetate (I-32)

[0828]

[0829] TFA (5 mL) was added dropwise to a stirred solution of F (542.00 mg, 1.185 mmol, 1.00 equiv) in DCM (10 mL) at room temperature. After stirring for 2 h, the resulting mixture was concentrated under reduced pressure, and the residue was purified by silica gel flash column chromatography to give I-32 (503 mg, 95.2%) as a yellow solid.1 1H NMR (300 MHz, DMSO-d6) δ 11.11 (s, 1H), 8.58 (d, 2H), 7.84 (dd, 1H), 7.49 (dd, 2H), 5.09 (dd, 1H), 4.24 (t, 2H), 4.10 – 3.84 (m, 5H), 3.11 – 2.80 (m, 2H), 2.66 - 2.53 (m, 1H), 2.16 - 1.98 (m, 3H). LCMS (ESI) m / z: [M+H] + = 358.13。

[0830] Preparation of 5-(azetidin-3-yloxy)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (I-33)

[0831]

[0832] Step 1: Preparation of tert-butyl 3-[(4-methylbenzenesulfonyl)oxy]azetidine-1-carboxylate (B)

[0833]

[0834] At 0 °C, DMAP (264.49 mg, 2.165 mmol, 0.15 equiv) and TEA (4.38 g, 43.300 mmol, 3.00 equiv) were added portionwise to a stirred solution of tert-butyl 3-hydroxyazetidine-1-carboxylate (A, 2.50 g, 14.433 mmol, 1.00 equiv) and p-toluenesulfonyl chloride (4.13 g, 21.650 mmol, 1.50 equiv) in DCM. The resulting mixture was concentrated under reduced pressure and the residue was purified by silica gel column chromatography, eluting with petroleum ether / EtOAc (1:1) to afford B (4.4 g, 93.11%) as a brown oil. LCMS (ESI) m / z: [M+H] + = 328。

[0835] Step 2: Preparation of tert-butyl 3-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]azetidine-1-carboxylate (C) Preparation of tert-butyl 3-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]azetidine-1-carboxylate (C)

[0836]

[0837] To a stirred solution of B (4.40 g, 13.439 mmol, 1.00 equiv) and KI (0.22 g, 1.344 mmol, 0.10 equiv) in DMF was added portionwise KHCO 3(4.04 g, 40.318 mmol, 3.00 equivalents). After stirring at 100 °C for 8 h, the resulting mixture was extracted with EtOAc (3 × 150 mL), and the organic extract was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, aqueous ACN solution, 10% to 50% gradient over 10 min; detector, UV 254 nm. This gave C (1.73 g, 29.98%) as an off-white solid. LCMS (ESI) m / z: [M+H] + = 430.

[0838] Step 3: Preparation of 5-(azetidin-3-yloxy)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione formate (I-33) Preparation of 5-(azetidin-3-yloxy)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione formate (I-33)

[0839]

[0840] A solution of C (1.53 g, 3.563 mmol, 1.00 equivalent) and TFA (5.00 mL, 67.315 mmol, 18.89 equivalents) in DCM was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure, and the residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, aqueous ACN solution, 10% to 50% gradient over 10 min; detector, UV 254 nm. This gave I-33 (1.08 g, 96.43%) as a white solid. 1 1H NMR (300 MHz, DMSO-d6) δ 11.11 (s, 1H), 8.25 (s, formic acid, 1H), 7.88 (d, 1H), 7.32 (d, 2H), 5.30 (p, 1H), 5.13 (dd, 1H), 4.31 (dd, 2H), 3.89 (dd, 2H), 2.99 - 2.80 (m, 1H), 2.68 – 2.52 (m, 2H), 2.13 - 1.97 (m, 1H). LCMS (ESI) m / z: [M+H] + = 330.05.

[0841] Preparation of 5-[2-(4-aminopiperidin-1-yl)ethoxy]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione formate (I-34)

[0842]

[0843] Step 1: Preparation of 5-(2-bromoethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (B) Preparation

[0844]

[0845] At 0 °C, to a solution of 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione (A, 1.37 g, 4.996 mmol, 1.00 equiv) in THF (35 mL) was added 2-bromoethanol (0.94 g, 7.494 mmol, 1.5 equiv), PPh 3 (1.97 g, 7.494 mmol, 1.5 equiv) and DIAD (1.52 g, 7.494 mmol, 1.5 equiv). The resulting mixture was stirred at room temperature for 2 h. After workup with water and extraction with DCM, the residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN / 0.1% aqueous formic acid, 0% to 100% gradient, over 45 min; detector, UV 254 nm. This gave B (1.52 g, 79.82%) as a black solid; LCMS (ESI) m / z: [M+H] + = 381, 383.

[0846] Step 2: Preparation of tert-butyl N-[1-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]ethyl)piperidin-4-yl]carbamate (C) Preparation of tert-butyl N-[1-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]ethyl)piperidin-4-yl]carbamate (C)

[0847]

[0848] To a solution of B (1.52 g, 3.988 mmol, 1.00 equiv) in ACN (35.00 mL) was added tert-butyl N-(piperidin-4-yl)carbamate (0.80 g, 3.988 mmol, 1.00 equiv), KI (0.66 g, 3.988 mmol, 1.00 equiv) and K 2 CO 3 (1.65 g, 11.963 mmol, 3.00 equiv). The resulting mixture was stirred at 70 °C for 2 h. After workup with water and extraction with DCM, the residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN / 0.1% aqueous formic acid, 0% to 100% gradient, over 30 min; detector, UV 254 nm. This gave C (1.402 g, 70.24%) as a colorless solid; LCMS (ESI) m / z: [M+H] + = 501.

[0849] Step 3: Preparation of 5-[2-(4-aminopiperidin-1-yl)ethoxy]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione formate (I-34) Preparation of 5-[2-(4-aminopiperidin-1-yl)ethoxy]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione formate (I-34)

[0850]

[0851] To a solution of C (1.66 g, 3.316 mmol, 1.00 equiv) in DCM (10.00 mL) was added TFA (10.00 mL, 134.630 mmol, 40.60 equiv). The resulting solution was stirred at room temperature for 3 h. After concentration, the residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN / 0.1% aqueous formic acid, 0% to 100% gradient, over 45 min; detector, UV 220 nm. This gave I-34 (840 mg, 62.52%) as a white solid; 1 H NMR (300 MHz, DMSO-d6) δ 11.13 (s, 1H), 8.52 - 8.26 (m, 2H), 8.16 (s, 1H, formic acid), 7.88 (d, 1H), 7.51 (d, 1H), 7.40 (dd, 1H), 5.14 (dd, 1H), 4.49 (t, 2H), 3.47 (d, 2H), 3.37 - 2.28 (m, 2H), 3.29 - 3.19 (m, 1H), 3.03 – 2.70 (m, 3H), 2.67 - 2.62 (m, 1H), 2.61 - 2.55 (m, 1H), 2.14 - 1.98 (m, 3H), 1.88 - 1.68 (m, 2H). LCMS (ESI) m / z: [M+H] + = 401.17.

[0852] Preparation of 5-[7-azaspiro[3.5]non-2-yloxy]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione formate (I-35)

[0853]

[0854] Step 1: Preparation of tert-butyl 2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]-7-azaspiro[3.5]nonane-7-carboxylate (B) Preparation of tert-butyl 2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]-7-azaspiro[3.5]nonane-7-carboxylate (B)

[0855]

[0856] To a solution of 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione (A, 1.37 g, 4.99 mmol, 1.00 equiv) and tert-butyl 2-hydroxy-7-azaspiro[3.5]nonane-7-carboxylate (1.81 g, 7.494 mmol, 1.5 equiv) in THF (30.00 mL) was added PPh 3 (1.97 g, 7.494 mmol, 1.5 equiv). DIAD (1.52 g, 7.494 mmol, 1.5 equiv) was added dropwise to the mixture over 10 minutes at 0 °C. The reaction mixture was stirred at room temperature for an additional 5 h. The resulting mixture was concentrated under reduced pressure and the residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN / 0.1% aqueous formic acid, 0% to 100% gradient over 45 min; detector, UV 254 nm. This gave B (1.964 g, 79.01%) as a white solid; LCMS (ESI) m / z: [M+H] + = 498.

[0857] Step 2: Preparation of 5-[7-azaspiro[3.5]non-2-yloxy]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione formate (I-35) Preparation of 5-[7-azaspiro[3.5]non-2-yloxy]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione formate (I-35)

[0858]

[0859] To a solution of B (1.96 g, 3.939 mmol, 1.00 equiv) in DCM (10.00 mL, 157.300 mmol, 39.93 equiv) was added TFA (10.00 mL, 134.630 mmol, 34.18 equiv). The resulting mixture was stirred at room temperature for 5 h, then concentrated and purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN / 0.1% aqueous formic acid, 0% to 100% gradient over 45 min; detector, UV 254 nm. This gave I-35 (1.6318 g, 93.13%) as a light gray solid: 11H NMR (300 MHz, DMSO-d6) δ 8.39 (s, formic acid, 1H), 7.84 (d, 1H), 7.35 – 7.24 (m, 2H), 5.12 (dd, 1H), 5.00 (p, 1H), 3.00 – 2.81 (m, 5H), 2.67 – 2.43 (m, 2H), 2.51 - 2.43 (m, 3H), 2.16 - 1.95 (m, 1H), 1.95 – 1.82 (m, 2H), 1.65 - 1.78 (m, 4H); LCMS (ESI) m / z: [M+H] + = 398.16。

[0860] Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-[2-(piperazin-1-yl)ethoxy]isoindole-1,3-dione formate (I-36)

[0861]

[0862] Step 1: Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-(prop-2-en-1-yloxy)isoindole-1,3-dione (B) Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-(prop-2-en-1-yloxy)isoindole-1,3-dione (B)

[0863]

[0864] To a solution of 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindole-1,3-dione (A, 5.48 g, 19.983 mmol, 1.00 equiv) and allyl bromide (3.63 g, 29.975 mmol, 1.5 equiv) in DMF (50.00 mL) was added KI (331.72 mg, 1.998 mmol, 0.1 equiv) and KHCO 3 3 (3.00 g, 29.975 mmol, 1.5 equiv). The resulting mixture was stirred at 65 °C for 12 h and then diluted with water (100 mL). The resulting mixture was extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with brine (3 x 60 mL) and dried over anhydrous Na 2 2 4 2SO4. After filtration, the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography, eluting with hexane / EtOAc (1:1), to give B (6.7 g, crude product) as a yellowish green solid; LCMS (ESI) m / z: [M+H] + = 315。

[0865] Step 2: 2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]acetaldehyde (C) Preparation

[0866]

[0867] To a solution of B (3.14 g, 9.991 mmol, 1.00 equiv) in 1,4-dioxane (30.00 mL) was added NaIO 4 (10.68 g, 49.953 mmol, 5.00 equiv), water (3.00 mL), and 2,6-lutidine (3.21 g, 29.972 mmol, 3 equiv). To the above mixture was added K 2 OsO 4 (0.37 g, 0.999 mmol, 0.1 equiv) at room temperature. K2OsO4.dihydrate (0.37 g, 0.999 mmol, 0.1 equiv) was added to the above mixture at room temperature. The resulting mixture was stirred at room temperature for an additional 2 h and then quenched with water. The resulting mixture was extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with brine (3 x 100 mL) and dried over anhydrous Na 2 SO 4 2. After filtration, the filtrate was concentrated under reduced pressure. This gave C (1.83 g, 57.92%) as a light brown solid; LCMS (ESI) m / z: [M+H] + = 317.

[0868] Step 3: Preparation of tert-butyl 4-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]ethyl)piperazine-1-carboxylate (D) Step 4: Preparation of formate of 2-(2,6-dioxopiperidin-3-yl)-5-[2-(piperazin-1-yl)ethoxy]isoindole-1,3-dione (I-36)

[0869]

[0870] At room temperature, to a solution of C (628.00 mg, 1.986 mmol, 1.00 equiv) and tert-butyl piperazine-1-carboxylate (369.84 mg, 1.986 mmol, 1 equiv) in DMF (10.00 mL) was added NaBH(OAc) 3 (1262.52 mg, 5.957 mmol, 3 equiv). The resulting mixture was stirred at room temperature for 3 h, then quenched with water, extracted into DCM and concentrated. The residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN / 0.1% aqueous formic acid, 0% to 100% gradient, over 45 min; detector, UV 254 nm. This gave D (812 mg, 84.05%) as an off-white solid; LCMS (ESI) m / z: [M+H] + = 487.

[0871] Step 1: Preparation of 5-(3-bromopropoxy)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (B) Step 2: Preparation of tert-butyl 4-(3-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]propyl)piperazine-1-carboxylate (C)

[0872]

[0873] To a solution of tert-butyl 4-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]ethyl)piperazine-1-carboxylate (D, 2.10 g, 4.316 mmol, 1.00 eq) in DCM (10.00 mL, 157.300 mmol, 36.44 eq) was added TFA (10.00 mL, 134.630 mmol, 31.19 eq). The resulting mixture was stirred at room temperature for 3 h and then concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN / 0.1% aqueous formic acid, 0% to 100% gradient, over 45 min; detector, UV 254 nm. This gave I-36 as a white solid (1.43 g, 74.24%); 1 H NMR (300 MHz, DMSO-d6) δ 11.07 (s, 1H), 8.34 (s, 1H), 7.84 (d, 1H), 7.47 (d, 1H), 7.37 (dd, 1H), 5.12 (dd, 1H), 4.30 (t, 2H), 2.99 - 2.86 (m, 5H), 2.77 (t, 2H), 2.67 - 2.55 (m, 5H), 2.13 - 1.96 (m, 1H).; LCMS (ESI) m / z: [M+H] + = 387.16.

[0874] Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-[3-(piperazin-1-yl)propoxy]isoindole-1,3-dione formate (I-37)

[0875]

[0876] Step 3: Preparation of formate of 2-(2,6-dioxopiperidin-3-yl)-5-[3-(piperazin-1-yl)propoxy]isoindole-1,3-dione (I-37) Step 1: Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-(pent-4-en-1-yloxy)isoindole-1,3-dione (B)

[0877]

[0878] To a solution of 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione (A, 1.37 g, 4.996 mmol, 1.00 equiv) and 3-bromopropan-1-ol (1.04 g, 7.494 mmol, 1.5 equiv) in THF (20.00 mL) was added PPh 3 (1.97 g, 7.494 mmol, 1.5 equiv). DIAD (1.52 g, 7.494 mmol, 1.5 equiv) was added dropwise thereto over 10 min at 0 °C. The resulting mixture was stirred at room temperature for an additional 5 h. After workup with water and extraction with DCM, the organic layer 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 / aqueous 0.1% formic acid, 0% to 100% gradient, over 45 min; detector, UV 254 nm. This gave B (1.85 g, 93.70%) as a colorless solid; LCMS (ESI) m / z: [M+H] + = 395, 397.

[0879] Step 2: Preparation of 4-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]butanal (C) Step 3: Preparation of tert-butyl N-[1-(4-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]butyl)piperidin-4-yl]carbamate (D)

[0880]

[0881] To a solution of B (1.85 g, 4.681 mmol, 1.00 equiv) and tert-butyl piperazine-1-carboxylate (1.05 g, 5.617 mmol, 1.2 equiv) in DMF (30.00 mL) were added DIEA (3.02 g, 23.405 mmol, 5 equiv) and KI (77.71 mg, 0.468 mmol, 0.1 equiv). The resulting mixture was stirred at 60 °C for 5 h. After workup and extraction, the residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN / aqueous 0.1% formic acid, 0% to 100% gradient, over 45 min; detector, UV 254 nm. This gave C (975 mg, 41.61%) as an off-white solid; LCMS (ESI) m / z: [M+H] + = 501.

[0882] Step 4: Preparation of 5-[4-(4-aminopiperidin-1-yl)butoxy]-2-(2,6-dioxopiperidin-3-yl)isoindole-1, 3-dione (

[0883]

[0884] To a solution of C (975.00 mg, 1.948 mmol, 1.00 equiv) in DCM (5.00 mL, 78.650 mmol, 40.38 equiv) was added TFA (5.00 mL, 67.315 mmol, 34.56 equiv). The resulting mixture was stirred at room temperature for 5 h. After concentration, the residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN / 0.1% aqueous formic acid, 0% to 100% gradient, over 45 min; detector, UV 254 nm. This gave I-37 (667 mg, 74.17%) as a white solid; 1 H NMR (300 MHz, DMSO-d6) δ 11.12 (s, 1H), 8.29 (d, formic acid, 1H), 7.84 (d, 1H), 7.44 (d, 1H), 7.36 (dd, 1H), 5.12 (dd, 1H), 4.23 (t, 2H), 3.17 – 2.85 (m, 5H), 2.67 – 2.58 (m, 2H), 2.53 - 2.57 (m, 4H), 2.50 – 2.29 (m, 3H), 2.15 - 2.05 (m, 1H), 2.01 - 1.82 (m, 2H); LCMS (ESI) m / z: [M+H] + = 401.17.

[0885] Preparation of 5-[4-(4-aminopiperidin-1-yl)butoxy]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (I-38)

[0886]

[0887] ) Step 1: Preparation of tert-butyl 4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]piperazine-1-carboxylate (B)

[0888]

[0889] To 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindole-1,3-dione (A, 500.00 mg, 1.823 mmol, 1.00 equiv) and KHCO 3A stirred solution of [[ID=]], (273.81 mg, 2.735 mmol, 1.50 equiv) in DMF (10.00 mL) was added portionwise with 5-bromo-1-pentene (326.07 mg, 2.188 mmol, 1.20 equiv) and KI (30.27 mg, 0.182 mmol, 0.10 equiv). The resulting mixture was stirred overnight at 65 °C under a nitrogen atmosphere. The mixture was cooled to room temperature and the resulting mixture was extracted with EtOAc (2 × 20 mL). The combined organic layers were washed with water (3 × 10 mL) and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure and the residue was purified by preparative TLC (petroleum ether / EtOAc 1:1) to afford B (400 mg, 64.08%) as a white solid. LCMS (ESI) m / z: [M+H] + = 343.

[0890] Step 2: Preparation of formate of 2-(2,6-dioxopiperidin-3-yl)-5-(piperazin-1-yl)isoindole-1,3-dione (I- 39)

[0891]

[0892] To a stirred solution of 2-(2,6-dioxopiperidin-3-yl)-5-(pent-4-en-1-yloxy)isoindoline-1,3-dione (B, 300.00 mg, 0.876 mmol, 1.00 equiv) in THF (1.00 mL), water (5.00 mL) and t-BuOH (5.00 mL) at room temperature was added portionwise with NMO (153.98 mg, 1.314 mmol, 1.50 equiv) and K 2 OsO 4 (32.29 mg, 0.088 mmol, 0.10 equiv). The resulting mixture was stirred overnight at room temperature under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. To the above mixture was added NaIO 4 (374.86 mg, 1.753 mmol, 2.00 equiv), water (0.50 mL) and acetone (5.00 mL). The reaction mixture was stirred at room temperature for an additional 2 h. After extraction with DCM, the resulting solution was concentrated under reduced pressure to afford C (300 mg, 99.43%) as an off-white solid. The crude product was used directly in the next step without further purification. LCMS (ESI) m / z: [M+H] + = 345.

[0893] Step 3: Preparation of tert-butyl 3-([4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]piperazin-1- yl]methyl)azetidine-1-carboxylate (C)

[0894]

[0895] At room temperature, STAB (249.29 mg, 1.176 mmol, 1.50 equiv) and tert-butyl N-(piperidin-4-yl)carbamate (157.05 mg, 0.784 mmol, 1.00 equiv) were added portionwise to a stirred solution of 4-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]butyraldehyde (C, 270.00 mg, 0.784 mmol, 1.00 equiv) in DMF (5.00 mL). The resulting mixture was stirred at room temperature overnight and then extracted with EtOAc (2 × 20 mL). The combined organic layers were washed with water (3 × 10 mL) and dried over anhydrous Na 2 SO 4 2SO4. After filtration, the filtrate was concentrated under reduced pressure and the residue was purified by preparative TLC (DCM / MeOH 10:1) to give D (130 mg, 31.36%) as an off-white solid. LCMS (ESI) m / z: [M+H] + = 529.

[0896] Step 4: Preparation of formate of 5-[4-(azetidin-3-ylmethyl)piperazin-1-yl]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (I-40) Step 1: Preparation of tert-butyl 3-[4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]piperazin-1-yl] I-38 propionate (B)

[0897]

[0898] At room temperature, tert-butyl N-[1-(4-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]butyl)piperidin-4-yl]carbamate (D, 120.00 mg, 0.227 mmol, 1.00 equiv), TFA (1.00 mL), and DCM (4.00 mL) were added to an 8 mL vial. The resulting mixture was stirred at room temperature for 2 h and then concentrated under reduced pressure. I-38 (130 mg, 93.55%) was obtained as an off-white solid. LCMS (ESI) m / z: [M+H] + = 429.

[0899] Preparation of 5-[4-(azetidin-3-ylmethyl)piperazin-1-yl]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione formate (I-40) and 2-(2,6-dioxopiperidin-3-yl)-5-(piperazin-1-yl)isoindole-1,3-dione formate (I-39)

[0900]

[0901] Step 2: Preparation of 3-[4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]piperazin-1- yl]propionic acid (I-41)

[0902]

[0903] To a solution of 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (A, 5.40 g, 19.549 mmol, 1.00 equiv) and tert-butyl piperazine-1-carboxylate (3.64 g, 19.549 mmol, 1 equiv) in DMF (35.00 mL) was added DIEA (7.58 g, 58.648 mmol, 3 equiv). The resulting mixture was stirred at 90 °C for 5 h. The reaction was quenched by the addition of water (50 mL) at room temperature. The resulting mixture was extracted with EtOAc (3 × 50 mL), and the combined organic layers were washed with brine (3 × 100 mL) and dried over anhydrous Na 2 SO 4 2SO4. After filtration, the filtrate was concentrated under reduced pressure. This gave B (7.32 g, 84.62%) as a pale yellow solid; LCMS (ESI) m / z: [M+H] + = 443.

[0904] ​ ​

[0905]

[0906] To a solution of B (900.00 mg, 2.034 mmol, 1.00 equiv) in DCM was added TFA (5.00 mL, 67.315 mmol, 33.09 equiv). The resulting mixture was stirred at room temperature for 3 h and then concentrated in vacuo. The residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN / 0.1% aqueous formic acid, 0% to 100% gradient over 45 min; detector, UV 254 nm. This gave I-39 (786.8 mg, 99.41%) as a yellow solid; LCMS (ESI) m / z: [M+H] + = 343.

[0907] ​ ​

[0908]

[0909] To a solution of I-39 (1.10 g, 3.213 mmol, 1.00 equiv) and tert-butyl 3-formylazetidine-1-carboxylate (595.13 mg, 3.213 mmol, 1 equiv) in DMF (10.00 mL) was added NaBH(OAc) 3 (2.04 g, 9.639 mmol, 3 equiv). The resulting mixture was stirred at room temperature for 3 h. After workup with water, extraction with DCM and concentration, the residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN / 0.1% aqueous formic acid, 0% to 100% gradient, over 45 min; detector, UV 254 nm. This gave C (991 mg, 60.29%) as a pale yellow solid; LCMS (ESI) m / z: [M+H] + = 512.

[0910] ​ ​

[0911]

[0912] To a solution of C (991.00 mg, 1.937 mmol, 1.00 equiv) in DCM (10.00 mL, 157.300 mmol, 81.20 equiv) was added TFA (10.00 mL, 134.630 mmol, 69.50 equiv). The resulting mixture was stirred at room temperature for 3 h and then concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN / 0.1% aqueous formic acid, 0% to 100% gradient, over 45 min; detector, UV 254 nm. This gave I-40 (702 mg, 85.29%) as a yellow solid; 1 1H NMR (300 MHz, DMSO-d6) δ 11.07 (s, 1H), 8.25 (s, formic acid, 1H), 7.69 (d, 1H), 7.34 (d, 1H), 7.26 (dd, 1H), 5.08 (dd, 1H), 4.00 (t, 2H), 3.66 (dd, 2H), 3.43 (t, 4H), 3.11 - 2.96 (m, 1H), 2.95 – 2.79 (m, 1H), 2.65 – 2.51 (m, 4H), 2.50 - 2.44 (m, 4H), 2.09 - 1.96 (m, 1H).; LCMS (ESI) m / z: [M+H]+ = 412.19。

[0913] Preparation of 3-[4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]piperazin-1-yl]propanoic acid (I-41)

[0914]

[0915] ​ ​

[0916]

[0917] To a solution of 2-(2,6-dioxopiperidin-3-yl)-5-(piperazin-1-yl)isoindole-1,3-dione (A, 5.0 g, 14.6 mmol, 1.0 equiv) and tert-butyl 3-bromopropanoate (3.6 g, 17.2 mmol, 1.2 equiv) in DMF (50 mL) was added dropwise DIEA (2.8 g, 21.7 mmol, 1.5 equiv), and the mixture was stirred at 70 °C for 6 h. The reaction mixture was concentrated, and the residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 spherical column; mobile phase, aqueous ACN solution, 10% to 70% gradient over 50 min; detector, UV 254 nm. tert-Butyl 3-[4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]piperazin-1-yl]propanoate (B) was obtained as a yellow solid. LCMS (ESI) m / z [M+H] + = 471。

[0918] ​ ​

[0919]

[0920] A solution of B (2.1 g, 4.5 mmol) in DCM (20 mL) and TFA (5 mL) was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure, and the residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 spherical column; mobile phase, aqueous MeOH solution, 10% to 30% gradient over 50 min; detector UV 254 nm to give 3-[4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]piperazin-1-yl]propanoic acid (I-41) (0.78 g, 42.2%) as a yellow solid. 11H NMR (300 MHz, DMSO-d6) δ 12.03 (br, 1H), 11.10 (s, 1H), 7.68 (d, J = 8.5 Hz, 1H), 7.35 (d, J = 1.7 Hz, 1H), 7.27 (dd, J = 8.6, 2.0 Hz, 1H), 5.08 (dd, J = 12.8, 5.4 Hz, 1H), 3.49–3.40 (m, 4H), 2.96–2.80 (m, 1H), 2.67–2.52 (m, 8H), 2.44 (t, J = 6.9 Hz, 2H), 2.07–1.97 (m, 1H). LCMS(ESI) m / z [M+H] + = 415.10。

[0921] Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-(4-(piperidin-4-ylmethyl)piperazin-1-yl)isoindoline-1,3-dione trifluoroacetate (I-42)

[0922]

[0923] Step 1: Preparation of tert-butyl 4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin- 1-yl)methyl)piperidine-1-carboxylate (B)

[0924]

[0925] A solution of 2-(2,6-dioxopiperidin-3-yl)-5-(piperazin-1-yl)isoindoline-1,3-dione (A, 1.5 g, 4.4 mmol, 1.0 equiv) and tert-butyl 4-formylpiperidine-1-carboxylate (0.93 g, 4.4 mmol, 1.0 equiv) in DMF (20 mL) was stirred at room temperature for 30 minutes. Then NaBH(OAc) 3 (3.7 g, 17.5 mmol, 4.0 equiv) was added and the resulting mixture was stirred at 50 °C for 3.5 h. The mixture was cooled to room temperature, poured into water (100 mL), and extracted with EtOAc (50 mL × 3). The combined organic layers were concentrated and the residue was purified by preparative HPLC under the following conditions: C18 spherical column, 20 - 35 um, 330 g; mobile phase, phase A: water (0.16% NH 4 HCO 3), B: ACN (gradient B% 0% - 70%, run time 40 minutes); flow rate: 80 mL / min; detector, UV detection at 254 nm. Obtained B as a yellow solid from purification (1.2 g, 50.8%). LCMS (ESI) m / z: [M+H] + = 540.

[0926] Step 2: Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-(4-(piperidin-4-ylmethyl)piperazin-1-yl)isoindole- 1,3-dione trifluoroacetate (I-42)

[0927]

[0928] A solution of tert-butyl 4-([4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]piperazin-1-yl]methyl)piperidine-1-carboxylate (B, 1.2 g, 2.2 mmol, 1.0 equiv) in DCM (8 mL) and TFA (2.0 mL) was stirred at room temperature for 2 h. The mixture was concentrated and the residue was purified by preparative HPLC under the following conditions: C18 spherical column, 20 - 35 um, 330 g; mobile phase, phase A: water (0.16% NH 4 HCO 3 ), B: ACN (gradient B% 0% - 60%, run time 45 minutes); flow rate: 80 mL / min; detector, UV detection at 254 nm. Obtained compound I-42 as a yellow solid (0.60 g, 61.3%). 1 H NMR (300 MHz, DMSO-d6): δ 10.42 (br, 1H), 9.12 (br,1H), 7.73–7.60 (m,1H), 7.35 (s, 1H), 7.26 (d, J = 8.7 Hz, 1H), 5.07 (dd, J =12.7, 5.3 Hz, 1H), 4.66 (br,1H), 3.12–3.33 (m, 5H), 2.93–2.75 (m, 5H), 2.19(d, J = 5.6 Hz, 3H), 2.07–1.98 (m, 2H), 1.85 (d, J = 11.7 Hz, 5H), 1.26 (d, J= 11.9 Hz, 3H). LCMS (ESI) m / z: [M+H] + = 440.35.

[0929] Preparation of 5-((2-aminoethyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione trifluoroacetate (I-43)

[0930]

[0931] Step 1: Preparation of 5-fluoroisobenzofuran-1,3-dione (B)

[0932]

[0933] Acetic anhydride (200 mL) was added to a solution of 4-fluorobenzene-1,2-dicarboxylic acid (A, 41.4 g, 0.22 mol, 1.00 equiv.). The reaction mixture was stirred at 120 °C for 2 h, then cooled to room temperature and concentrated under reduced pressure to give 5-fluoro-2-benzofuran-1,3-dione (B, 37.2 g, quantitative) as a white solid.

[0934] Step 2: Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindole-1,3-dione (C)

[0935]

[0936] Under a nitrogen atmosphere, NaOAc (55.2 g, 0.67 mol, 3.0 equiv.) was added to a stirred solution of 5-fluoro-2-benzofuran-1,3-dione (B, 37.2 g, 0.22 mol, 1.0 equiv.) and 3-aminopiperidine-2,6-dione hydrochloride (55.4 g, 0.34 mol, 1.5 equiv.) in acetic acid (300 mL). The resulting mixture was stirred at 120 °C for 3 h, then cooled to room temperature. The suspension was filtered and the cake was washed with water (3 x 150 mL) to give 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindole-1,3-dione (C, 53.1 g, 87.4%) as a grey solid. LCMS (ESI) m / z: [M+H] + = 277.

[0937] Step 3: Preparation of tert-butyl (2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)ethyl)carbamate (D) Step 4: Preparation of 5-((2-aminoethyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione trifluoroacetate (I-43)

[0938]

[0939] To a solution of tert-butyl N-(2-aminoethyl)carbamate (2.9 g, 18.1 mmol, 1.0 equiv) and 2-(2,6-dioxopiperidin-3-yl)-5-fluoro-1H-isoindole-1,3-dione (C, 5.0 g, 18.1 mmol, 1.0 equiv) in DMF (50 mL) was added DIEA (4.7 g, 36.2 mmol, 2.00 equiv). The mixture was stirred at 80 °C for 2 h, then cooled to room temperature and poured into water (500 mL). The resulting mixture was extracted with EtOAc (500 mL x 3). The combined organic layers were concentrated and the residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 spherical column; mobile phase, aqueous ACN solution, 0% to 100% gradient, over 50 min; 70 ml / min detector, UV 254 nm, to afford the desired product tert-butyl N-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]amino)ethyl)carbamate (D, 1.5 g, 19.5%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 417。

[0940] Step 1: Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindole-1,3-dione (B) Step 2: Preparation of tert-butyl (2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethyl)carbamate (C)

[0941]

[0942] At 0 °C, trifluoroacetic acid (4 mL) was added dropwise to a stirred solution of tert-butyl N-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]amino]ethyl)carbamate (D, 1.5 g, 3.6 mmol, 1.0 equiv) in DCM (20 mL). The resulting mixture was warmed to room temperature and stirred for 2 h. The reaction mixture was concentrated under reduced pressure and the crude product was purified by preparative HPLC to afford 5-[(2-aminoethyl)amino]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione trifluoroacetate (I-43, 0.97 g, 63.1%) as a yellow solid. 11H NMR (300 MHz, DMSO-d6) δ 11.09 (s, 1H), 7.88 (br, 3H), 7.63 (d, J = 8.3 Hz, 1H), 7.22 (br, 1H), 7.04 (d, J = 1.9 Hz, 1H), 6.92 (dd, J = 8.4, 2.1 Hz, 1H), 5.06 (dd, J = 12.8, 5.3 Hz, 1H), 3.52–3.39 (m, 2H), 3.08–2.81 (m, 3H), 2.65–2.53 (m, 2H), 2.06–1.96 (m, 1H). LCMS (ESI) m / z: [M+H] + = 317.00。

[0943] Preparation of 5-(2-aminoethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione trifluoroacetate (I-44)

[0944] Step 3:

[0945]

[0946] To a stirred solution of 5-hydroxy-2-benzofuran-1,3-dione (A, 2.0 g, 12.2 mmol, 1.0 equiv) and 3-aminopiperidine-2,6-dione hydrochloride (3.0 g, 18.3 mmol, 1.5 equiv) in HOAc (40 mL) was added NaOAc (3.0 g, 36.6 mmol, 3.0 equiv). The reaction mixture was stirred at 120 °C under a nitrogen atmosphere and then concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography, eluting with petroleum ether / EtOAc (4:1) to afford B (2.9 g, 86.7%) as a white solid. LCMS (ESI) m / z [M+H] + = 275。

[0947] - Preparation of 2-(2-aminoethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione trifluoroacetate (I-44) Step 1: Preparation of tert-butyl (2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)ethyl)carbamate (B)

[0948]

[0949] Under a nitrogen atmosphere, DIEA (0.94 g, 7.3 mmol, 2.0 equiv) was added to a stirred solution of B (1.0 g, 3.6 mmol, 1.0 equiv) and tert-butyl 1,2,3-oxathiazolidine-3-carboxylate 2,2-dioxide (0.98 g, 4.4 mmol, 1.2 equiv) in DMF (10 mL). After stirring at 80 °C for 2 h, water (100 mL) was added, and then the mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with water (3×100 mL) and dried over anhydrous Na 2 SO 4 4. The filtrate was concentrated to dryness under reduced pressure, and the residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 spherical column; mobile phase, aqueous ACN solution, 0% to 100% gradient over 50 min; flow rate, 70 ml / min; detector, UV 254 nm, to give C (1.7 g, 94.4%) as a yellow oil. LCMS (ESI) m / z: [M+H] + = 418.

[0950] Step 2: Preparation of 3-(4-((2-aminoethyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I- 5 45) Step 1: Preparation of methyl 2-(bromomethyl)-5-nitrobenzoate (B)

[0951]

[0952] TFA (5 mL) was added dropwise to a stirred solution of C (1.4 g, 3.4 mmol, 1.0 equiv) in DCM (20 mL). After stirring at room temperature for 2 h, the reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel flash column chromatography to give I-44 (750 mg, 51.2%) as a yellow solid. 1 1H NMR (300 MHz, DMSO-d6) δ11.15 (s, 1H), 8.04 (br, 3H), 7.91 (d, J = 8.3 Hz, 1H), 7.50 (d, J = 2.2 Hz,1H), 7.41 (dd, J = 8.3, 2.3 Hz, 1H), 5.14 (dd, J = 12.9, 5.3 Hz, 1H), 4.38(t, J = 4.9 Hz, 2H), 3.35–3.23 (m, 2H), 2.98–2.81 (m, 1H), 2.66–2.54 (m, 2H),2.11–2.01 (m, 1H).LCMS (ESI) m / z: [M+H] += 418.10。

[0953] Preparation of 3-(4-((2-aminoethyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-45)

[0954]

[0955] Step 2: Preparation of 3-(6-nitro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (C) Step 3: Preparation of 3-(6-amino-1-oxoisoindolin-2-yl)piperidine-2,6-dione (D)

[0956]

[0957] Lenalidomide (A, 442.00 mg, 1.705 mmol, 1.00 equiv), N-(2-oxoethyl)carbamic acid tert-butyl ester (229.00 mg, 1.439 mmol, 0.84 equiv) and AcOH (100.00 mg, 1.665 mmol, 0.98 equiv) were suspended in MeOH (20.00 mL). The mixture was stirred at room temperature for 2 h, then NaBH3CN (145.00 mg, 2.307 mmol, 1.35 equiv) was added. After 1 h, the reaction was quenched with saturated aqueous NaHCO3 and extracted with DCM (3 × 40 mL). The combined organic layers were washed with brine (1 × 100 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 DCM:MeOH (10:1) to give B (100 mg, 14.58%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 403。

[0958] Step 4: Preparation of tert-butyl (2-((2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindol-5-yl)amino)ethyl)carbamate (E) Step 5: Preparation of 3-(6-((2-aminoethyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-

[0959]

[0960] B (100.00 mg, 0.248 mmol, 1.00 equiv) was dissolved in DCM (5.00 mL). TFA (1.00 mL, 13.463 mmol, 54.18 equiv) was added and the resulting solution was stirred at room temperature for 0.5 h. The solution was concentrated under reduced pressure to give I-45 as a crude product, which was used in the next step without further purification. LCMS (ESI) m / z: [M+H] + =303。

[0961] Preparation of 3-(6-((2-aminoethyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-46)

[0962]

[0963] 46)

[0964]

[0965] Dissolve A (2.00 g, 10.247 mmol, 1.00 equiv), NBS (2.40 g, 13.484 mmol, 1.32 equiv) and BPO (80.00 mg, 0.312 mmol, 0.03 equiv) in CCl4 (40.00 mL). Heat the solution to reflux and stir overnight. Wash the resulting mixture with 50 mL of brine, and purify the residue by silica gel column chromatography, eluting with petroleum ether:EtOAc (5:1) to give B (1.2 g, 42.73%) as a colorless oil. LCMS (ESI) m / z: [M+H] + = 275.

[0966] Step 1: Preparation of methyl 2-(bromomethyl)-4-nitrobenzoate (B)

[0967]

[0968] Dissolve B (510.00 mg, 1.861 mmol, 1.00 equiv), 3-aminopiperidine-2,6-dione (395.00 mg, 3.083 mmol, 1.66 equiv) and DIEA (777.00 mg, 6.012 mmol, 3.23 equiv) in ACN (10.00 mL). Stir the mixture at 80 °C for 2 h. Concentrate the reaction mixture, and purify the residue by silica gel column chromatography, eluting with DCM:ACN (3:1) to give C (410 mg, 76.17%) as a dark green solid. LCMS (ESI) m / z: [M+H] + = 290.

[0969] Step 2: Preparation of 3-(5-nitro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (C)

[0970]

[0971] C (410.00 mg, 1.417 mmol, 1.00 equivalent) and 10% Pd / C (80.00 mg, 0.752 mmol, 0.53 equivalent) were suspended in DMF (15.00 mL). The reaction mixture was stirred overnight at room temperature and 1 atm of hydrogen. The mixture was filtered through Celite, and the filtrate was concentrated to give D (350 mg, 95.24%) as a brown solid. LCMS (ESI) m / z: [M+H] + = 260.

[0972] Step 3: Preparation of 3-(5-amino-1-oxoisoindolin-2-yl)piperidine-2,6-dione (D) ​

[0973]

[0974] D (350.00 mg, 1.350 mmol, 1.00 equivalent), tert-butyl N-(2-oxoethyl)carbamate (257.87 mg, 1.620 mmol, 1.20 equivalents), and AcOH (81.07 mg, 1.350 mmol, 1.00 equivalent) were suspended in MeOH (15.00 mL). The mixture was stirred at room temperature for 1 h, then NaBH3CN (101.80 mg, 1.620 mmol, 1.20 equivalents) was added. The reaction was stirred for an additional 2 h and then quenched with 5% aqueous Na2HCO3. The resulting mixture was extracted with DCM (40 mL x 3), and the combined organic layers were dried over Na2SO4 and concentrated to give E (450 mg, 82.83%) as a white solid. LCMS (ESI) m / z: [M+H] + = 403.

[0975] ​ ​

[0976]

[0977] E (50.00 mg, 0.124 mmol, 1.00 equivalent) and TFA (0.50 mL, 6.732 mmol, 54.18 equivalents) were dissolved in DCM (2.00 mL, 31.460 mmol, 253.22 equivalents). The solution was stirred at room temperature for 0.5 h and then concentrated to give I-46 (50 mg, crude product) as a yellow oil, which was used without further purification. LCMS (ESI) m / z: [M+H] + = 303.

[0978] Preparation of 3-(5-((2-aminoethyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-47)

[0979] ​

[0980]

[0981] Dissolve methyl 2-methyl-4-nitrobenzoate (A, 2.00 g, 1 equivalent), NBS (2.40 g) and BPO (88.00 mg) in CCl 4 (40.00 mL). Heat the solution to reflux and stir overnight. Wash the resulting mixture with 50 mL of brine, dry over Na 2 SO 4 and concentrate under reduced pressure. Purify the residue by silica gel column chromatography, eluting with petroleum ether:EtOAc (5:1) to give B (2.2 g, 78.33%) as a colorless oil. LCMS (ESI) m / z: [M+H] + = 275.

[0982] ​

[0983]

[0984] Suspend B (508.50 mg, 1.855 mmol, 1.00 equivalent), 3-aminopiperidine-2,6-dione (370.70 mg, 2.893 mmol, 1.56 equivalents) and DIEA (755.00 mg, 5.842 mmol, 3.15 equivalents) in ACN (10.00 mL). Stir the mixture at 80 °C for 2 h, then cool and concentrate under reduced pressure. Purify the residue by silica gel column chromatography, eluting with DCM:ACN (3:1) to give C (230 mg, 42.86%) as a gray solid. LCMS (ESI) m / z: [M+H] + = 290.

[0985] ​

[0986]

[0987] C (190.00 mg, 0.657 mmol, 1.00 eq) and 10% Pd / C (44 mg) were suspended in DMF (5.00 mL). The mixture was stirred overnight at room temperature and 1 atm of hydrogen. The reaction mixture was filtered through celite and concentrated to give D (170 mg, 99.82%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 260.

[0988] Step 4: Preparation of tert-butyl (2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)amino)ethyl)carbamate (E) Step 5: Preparation of 3-(5-((2-aminoethyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-

[0989]

[0990] D (250.00 mg, 0.964 mmol, 1.00 eq), N-(2-oxoethyl)carbamic acid tert-butyl ester (153.50 mg, 0.964 mmol, 1.00 eq) and AcOH (0.10 mL, 1.745 mmol, 1.81 eq) were suspended in MeOH (10.00 mL). The reaction mixture was stirred at room temperature for 1 h, then NaBH3CN (60.60 mg, 0.964 mmol, 1.00 eq) was added. The mixture was stirred for an additional 2 h and then diluted with saturated NaHCO 3 (20 mL). The aqueous layer was extracted with DCM (3×40 mL), and the combined organic layers were dried over Na 2 SO 4 dried, filtered and concentrated. The residue was purified by silica gel column chromatography, eluting with petroleum ether / EtOAc (1:1) to give E (100 mg, 25.77%) as a pale yellow solid. LCMS (ESI) m / z: [M+H] + = 403.

[0991] 47) Step 1: Preparation of ethyl 2-(2-(3-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-

[0992]

[0993] E (100.00 mg, 0.248 mmol, 1.00 eq) was dissolved in DCM (5.00 mL). TFA (1.00 mL, 13.463 mmol, 54.18 eq) was added and the resulting solution was stirred at room temperature for 0.5 h. The mixture was concentrated to give I-47 (100 mg, crude product) as a yellow oil. LCMS (ESI) m / z: [M+H] + = 303.

[0994] Preparation of 3-(6-((6-Aminohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-48) and 3-(5-((6-Aminohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-49)

[0995]

[0996] At room temperature, zinc powder (351.26 mg, 5.370 mmol, 10.00 equivalents) was added portionwise to a stirred solution of 5-[(6-aminohexyl)amino]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (A, 200.00 mg, 0.537 mmol, 1.00 equivalent) in HOAc (10.00 mL). The resulting mixture was stirred at 60 °C for 30 minutes. The resulting mixture was filtered and the filter cake was washed with EtOAc (4 × 10 mL). The filtrate was concentrated under reduced pressure and the residue was purified by preparative HPLC to give I-48 (19 mg, 9.87%) as a white solid and I-49 (23 mg, 11.95%) as a white solid. Respective LCMS (ESI) m / z: [M+H] + = 359.

[0997] Preparation of 7-[[(2S)-1-[(2S,4R)-4-Hydroxy-2-([[4-(4-Methyl-1,3-thiazol-5-yl)phenyl]methyl]carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamoyl]heptanoic acid (I-50)

[0998]

[0999] At 0 °C, A (1.00 g, 2.323 mmol, 1.00 equivalent), TEA (822.55 mg, 8.129 mmol, 3.50 equivalents), HOAt (347.73 mg, 2.555 mmol, 1.10 equivalents) and EDCI (489.75 mg, 2.555 mmol, 1.10 equivalents) were added to a stirred solution of suberic acid (2.02 g, 11.596 mmol, 4.99 equivalents) in DCM (25.00 mL) and THF (25.00 mL). The resulting solution was stirred at 0 °C for 2 hours. The resulting mixture was concentrated under reduced pressure and the residue was purified by reverse-phase flash chromatography to give I-50 (900 mg, 66.04%) as a white solid. LCMS (ESI) m / z: [M+H]+ = 587。

[1000] Preparation of 6-[[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]-carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamoyl]spiro[3.3]heptane-2-carboxylic acid (I-51)

[1001]

[1002] I-51 was prepared in a similar manner as described in the preparation of I-50. LCMS (ESI) m / z: [M+H] + = 597。

[1003] Preparation of 3-(2-(3-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)ethoxy)propanoic acid (I-52)

[1004]

[1005] yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropoxy) ethoxy)propionate (B) Step 2: Preparation of 2-(2-(3-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-

[1006]

[1007] To a solution of A (500 mg, 1.07 mmol, HCl), 3-[2-(3-ethoxy-3-oxopropoxy)ethoxy]propanoic acid (250.79 mg, 1.07 mmol, 250.79 µL) and DIEA (691.84 mg, 5.35 mmol, 932.40 µL) in DCM (5 mL) was added EDCI (246.29 mg, 1.28 mmol) and HOBt (173.60 mg, 1.28 mmol). The mixture was stirred at 20 °C for 16 h and then concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (0.1% aqueous formic acid / ACN) and then lyophilized to give B (500 mg, 70.00%) as a yellow oil. LCMS (ESI) m / z: [M+H] + = 647.6。

[1008] yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropoxy) ethoxy)propanoic acid (I-52) Step 1:

[1009]

[1010] A solution of NaOH (77.30 mg, 1.93 mmol) in water (1 mL) was added to a solution of B (500 mg, 0.7 mmol) in EtOH (5 mL). The mixture was stirred at 20 °C for 1 h, then diluted with water (30 mL) and adjusted to pH 6 with 1 N HCl. The mixture was extracted with EtOAc (30 mL × 3), and the combined organic layers were dried over Na 2 SO 4 and concentrated to give I-52 (470 mg, 95.02%) as a brown solid, which was used without further purification. LCMS (ESI) m / z: [M+Na] + = 641.2.

[1011]

[1012]

[1013]

[1014]

[1015] (2S,4R)-1-[(2S)-2-(2-[2-[2-(2-Aminoethoxy)ethoxy]ethoxy]acetamido)-3,3-dimethylbutanoyl]-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (I-70) Preparation

[1016]

[1017] Preparation of (B) tert-Butyl N-(2-[2-[2-([[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamoyl)methoxy]ethoxy]ethoxy)ethylcarbamate Step 2:

[1018]

[1019] To a stirred solution of (2S,4R)-1-[(2S)-2-amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (A, 1.00 g, 2.323 mmol, 1.00 equiv) and 2-[2-[2-(2-[[(tert-butoxy)carbonyl]amino]ethoxy)ethoxy]ethoxy]acetic acid (785.19 mg, 2.555 mmol, 1.10 equiv) in DCM (10.00 mL) was added HATU (1.32 g, 3.484 mmol, 1.50 equiv) and DIEA (900.50 mg, 6.968 mmol, 3 equiv). The mixture was stirred at 25 °C for 1 h and then diluted with EtOAc (200 mL) and washed with 3 x 100 mL water and 1 x 100 mL saturated brine. The organic layer was dried over Na 2 SO 4 4, filtered and evaporated. The crude product was purified by flash silica chromatography eluting with a gradient of 0 to 50% EtOAc / petroleum ether to afford B (1.4592 g, 87.27%) as an off-white solid. LCMS (ESI) m / z: [M+H] + = 720.

[1020] Preparation of (I-70) (2S,4R)-1-[(2S)-2-(2-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]acetamido)-3,3-dimethylbutanoyl]-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide Step 1: Preparation of 2-[2-[2-(2-benzyloxy-2-oxo-ethoxy)ethoxy]ethoxy]acetic acid (B)

[1021]

[1022] To a solution of B (1.45 g, 2.014 mmol, 1 equiv) in DCM (15 mL) was added dropwise TFA (3 mL). After stirring at room temperature for 2 h, the mixture was concentrated under reduced pressure and the residue was purified by flash column chromatography on silica gel to afford I-70 (1.05 g, 84.11%) as an off-white solid. 11H NMR (400 MHz, methanol-d4) δ 8.90 (s, 1H), 7.51 – 7.42 (m, 4H), 4.72 (s, 1H), 4.63 – 4.49 (m, 3H), 4.46 – 4.35 (m, 1H), 4.16 – 4.03 (m, 2H), 3.94 – 3.79 (m, 2H), 3.77 – 3.62 (m, 8H), 3.59 (t, J = 5.2 Hz, 2H), 2.99 – 2.87 (m, 2H), 2.50 (s, 3H), 2.30 – 2.21 (m, 1H), 2.19 – 2.06 (m, 1H), 1.13 – 0.98 (m, 9H). LCMS (ESI) m / z: [M+H] + = 620.30。

[1023]

[1024] (S)-13-((2S,4R)-4-Hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carbonyl)-14,14-dimethyl-11-oxo-3,6,9-trioxa-12-azapentadecanoic acid (I-75) Preparation

[1025]

[1026] Step 2: Preparation of benzyl (S)-13-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)

[1027]

[1028] At 0 °C, bromotoluene (2.42 g, 14.18 mmol, 1.68 mL) was added dropwise to a mixture of 2-[2-[2-(carboxymethoxy)ethoxy]ethoxy]acetic acid (A, 3 g, 13.50 mmol) and TEA (3.51 g, 34.71 mmol, 4.83 mL) in acetone (20 mL). The mixture was stirred at 20 °C for 16 h, during which a thick precipitate formed. The solid was filtered off and washed with acetone (10 mL). The filtrate was concentrated and the residue was dissolved in water (300 mL). The mixture was washed with EtOAc (50 mL x 3), and the aqueous layer was treated with HCl (2 M) to a final pH of 3 - 5. The mixture was extracted with EtOAc (50 mL x 3), and the combined organic layers were washed with brine (50 mL), over Na 2 SO 4It was dried and concentrated under vacuum to give B as a yellow oil (1.4 g, 4.48 mmol, 33.20% yield). The gas was used directly without further purification. 1 H NMR (400 MHz, CDCl 3 ) δ = 7.31 - 7.26 (m, 5H), 5.12 (s, 2H), 4.12 (s, 2H), 4.08 (s, 2H), 3.69 - 3.61(m, 8H) ppm.

[1029] pyrrolidine-1-carbonyl)-14,14-dimethyl-11-oxo-3,6,9-trioxa-12-azapentadecanoate (C) Step 3: Preparation of (S)-13-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl) pyrrolidine-1-carbonyl)-14,14-dimethyl-11-oxo-3,6,9-trioxa-12-azapentadecanoic acid (I-75)

[1030]

[1031] To B (836.17 mg, 2.68 mmol) in DCM (10 mL) was added HATU (1.32 g, 3.48 mmol) and DIEA (900.50 mg, 6.97 mmol, 1.21 mL). Then (2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[[4-(4-methylthiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (1 g, 2.14 mmol, HCl) was added. The mixture was stirred at 30 °C for 2 h. The mixture was concentrated under vacuum to give a yellow solid. The residue was purified by reverse phase flash (0.1% aqueous formic acid / ACN) and the fractions were lyophilized to give C as a yellow oil (1.2 g, 1.66 mmol, 71.28% yield).

[1032] LCMS (ESI) m / z: [M+H] + = 725.4. 1 H NMR (400 MHz, chloroform-d) δ = 8.69 (s,1H), 7.42 - 7.32 (m, 11H), 4.77 - 4.75 (m, 1H), 4.63 - 4.47 (m, 3H), 4.37 - 4.32 (m,1H), 4.24 - 4.17 (m, 3H), 4.13 (d, J= 11.6 Hz, 1H), 4.09 - 3.94 (m, 2H), 3.77 - 3.68 (m, 9H), 3.63 - 3.59 (m, 1H), 2.64 - 2.56 (m, 1H), 2.54 (s, 3H), 2.19 - 2.08(m, 1H), 1.00 - 0.92 (m, 9H) ppm。

[1033] Step 1: Preparation of (2S,4R)-1-[(2S)-2-(2-chloroacetamido)-3,3-dimethylbutanoyl]-4-hydroxy-N- [[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (B) Step 2:

[1034]

[1035] To a mixture of C (1.1 g, 1.52 mmol) in MeOH (20 mL) was added 10% Pd / C (500 mg). The mixture was purged with hydrogen, then pressurized to 15 psi and stirred at 25 °C for 12 h, then stirred at 40 °C for 8 h. The mixture was filtered and the filtrate was concentrated in vacuo. The residue was purified by reverse-phase flash (ammonia / water conditions) to give I-75 (560 mg, 882.25 μm, 58.14% yield) as a white solid. LCMS (ESI) m / z: [M+H] + = 635.2. 1 H NMR (400 MHz, methanol-d4) δ = 8.94 - 8.86 (m, 1H), 7.50 - 7.43 (m,4H), 4.62 - 4.50 (m, 3H), 4.43 - 4.34 (m, 1H), 4.12 (s, 2H), 4.08 (d, J = 5.2 Hz,2H), 3.93 - 3.86 (m, 1H), 3.85 - 3.79 (m, 1H), 3.76 - 3.70 (m, 8H), 2.51 - 2.49 (m,3H), 2.29 - 2.21 (m, 1H), 2.16 - 2.07 (m, 1H), 1.06 (s, 9H) ppm。

[1036]

[1037] Preparation of 2-([[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamoyl]methyl)-2-azaspiro[3.3]heptane-6-carboxylic acid (I-78)

[1038]

[1039] Preparation of (C) Step 3: Preparation of 2-([[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazol-5-yl)phenyl]

[1040]

[1041] At 0 °C, chloroacetyl chloride (137.71 mg, 1.219 mmol, 1.05 eq) was added dropwise to a solution of (2S,4R)-1-[(2S)-2-amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (A, 500.00 mg, 1.161 mmol, 1.00 eq) and DIEA (450.25 mg, 3.484 mmol, 3.00 eq) in DCM (10.00 mL, 0.118 mmol, 0.10 eq). The solution was stirred at room temperature for 4 h and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with DCM / MeOH (10:1) to give B (450 mg, 76.43%) as a yellow oil. LCMS (ESI) m / z: [M+H] + = 507.

[1042] methyl]carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamoyl]methyl)-2-aza Methyl 2-([[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamoyl]methyl)-2-azaspiro[3.3]heptane-6-carboxylate spiro[3.3]heptane-6-carboxylic acid (I-78)

[1043]

[1044] To methyl 2-azaspiro[3.3]heptane-6-carboxylate (24.00 mg, 0.155 mmol, 1.00 eq) and K 2 CO 3A suspension of A (64.12 mg, 0.464 mmol, 3.00 equiv) in DMF (3.00 mL) was added to B (78.41 mg, 0.155 mmol, 1 equiv). The suspension was stirred at room temperature for 16 h. After workup with water, extraction with DCM and concentration, the residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, aqueous ACN solution, 5% to 100% gradient over 15 min; detector, UV 254 nm. This gave C (30 mg, 31.00%) as a yellow oil. LCMS (ESI) m / z: [M+H] + = 626。

[1045] Step 1: 6-([[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazol-5-yl)phenyl] methyl]carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamoyl]methyl)-2,6-di ​

[1046]

[1047] To a solution of C (60.00 mg, 0.096 mmol, 1.00 equiv) in THF (3.00 mL) and water (1.00 mL) was added LiOH (5.74 mg, 0.240 mmol, 3.00 equiv). The solution was stirred at room temperature for 3 h and the mixture was acidified to pH 5 with acetic acid. The mixture was purified directly by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, aqueous ACN solution, 10% to 100% gradient over 15 min; detector, UV 254 nm. This gave I-78 (42 mg, 71.61%) as a white solid. LCMS (ESI) m / z: [M+H] + = 612。

[1048] Preparation of [6-([[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamoyl)methyl]-2,6-diazaspiro[3.3]hept-2-yl]acetic acid (I-79)

[1049]

[1050] ​ ​ Preparation of tert-Butyl Azaspiro[3.3]heptane-2-carboxylate (B)

[1051]

[1052] A (200.00 mg, 0.394 mmol, 1.00 equiv), K 2 CO 3 (136.28 mg, 0.986 mmol, 2.50 equiv), and tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate (156.41 mg, 0.789 mmol, 2.00 equiv) were added to DMF (5.00 mL). The mixture was reacted at room temperature for 24 h. The resulting mixture was diluted with EtOAc (30 mL), washed with water (20 mL) and brine (20 mL), and dried over Na 2 SO 4 . After removal of the organic solvent, the residue was purified by preparative TLC (5% MeOH / EtOAc) to give B (90 mg, 34.11%) as a white solid. LCMS (ESI) m / z: [M+H] + = 669.

[1053] Step 2: (2S,4R)-1-[(2S)-2-(2-[2,6-Diazaspiro[3.3]hept-2-yl]acetamido)-3,3- dimethylbutanoyl]-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (C)

[1054]

[1055] B (60.00 mg, 0.090 mmol, 1.00 equiv) was dissolved in DCM (5.00 mL), and then TFA (1.00 mL, 13.463 mmol, 150.08 equiv) was added. The mixture was reacted for 1 h. After removal of the solvent, the crude product C was used without further purification. LCMS (ESI) m / z: [M+H] + = 569.

[1056] Step 3: Preparation of tert-Butyl 2-[6-([[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazol-5-yl)benz yl]methyl]aminocarbonyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]aminocarbonyl]methyl)-2, 6-diazaspiro[3.3]heptane-2-carboxylate (D)

[1057]

[1058] To a suspension of C (41.00 mg, 0.072 mmol, 1.00 equiv) and K 2 CO 3 (29.89 mg, 0.216 mmol, 3.00 equiv) in DMF (1.50 mL) was added dropwise DMF (0.1 mL) intert-Butyl 2-chloroacetate (10.86 mg, 0.072 mmol, 1.00 equiv). The mixture was stirred at room temperature for 4.5 h. Then the reaction mixture was diluted with 30 mL of water and then extracted with DCM / MeOH (10 / 1, v / v, 20 mL x 3). The combined organic layers were washed with brine (40 mL) and dried over sodium sulfate. After filtration, the filtrate was concentrated in vacuo to give the crude product D (45 mg, 91.41%), which was used without further purification. LCMS (ESI) m / z: [M+H] + = 683。

[1059] Step 4: Preparation of [6-([[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazol-5-yl)benz yl]methyl]aminocarbonyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]aminocarbonyl]methyl)-2, 6-diazaspiro[3.3]heptane-2-carboxylic Acid (I-79)

[1060]

[1061] D (25.00 mg, 0.037 mmol, 1.00 equiv) was dissolved in DCM (5.00 mL), and then TFA (1.25 mL, 10.963 mmol, 459.68 equiv) was added. The mixture was stirred at room temperature for 4 h. After removal of the solvent, the crude product I-79 (23 mg, 100.24%) was used without further purification. LCMS (ESI) m / z: [M+H] + =627。

[1062] Preparation of 1-([[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamoyl]methyl)piperidine-4-carboxylic acid (I-80)

[1063]

[1064] Step 1: tert-Butyl 1-(2-ethoxy-2-oxoethyl)piperidine-4-carboxylate (Preparation of (B)

[1065]

[1066] To a solution of ethyl glyoxylate (500.00 mg, 4.898 mmol, 1.00 equiv) and tert-butyl piperidine-4-carboxylate (A, 1088.86 mg, 5.877 mmol, 1.20 equiv) in EtOH (10 mL) was added AcOH (29.41 mg, 0.49 mmol, 0.1 equiv), and the resulting solution was stirred at room temperature for 1 h. Then NaBH 3 CN (461.67 mg, 7.347 mmol, 1.50 equiv) was added, and the resulting solution was stirred at room temperature overnight. Then the reaction was quenched by the addition of 10 mL of water. The resulting solution was extracted with 3 × 30 mL of EtOAc, and the combined organic extracts were concentrated in vacuo. The residue was purified by flash column chromatography on silica gel using EtOAc / petroleum ether (1:3). This gave 200 mg (15.05%) of B as a yellow oil. 1 H NMR (300 MHz, DMSO-d 6 ) δ 4.07 (q, J = 7.1 Hz, 2H), 3.19 (s, 2H), 2.77 (d, J = 11.7 Hz, 2H), 2.27 – 2.07 (m, 3H), 1.74 (dd, J = 13.1, 3.7 Hz, 2H), 1.60 – 1.44 (m, 1H), 1.40 (s, 9H), 1.19 (t, J = 7.1 Hz, 3H). LCMS (ESI) m / z: [M+H] + = 272.

[1067] Step 2: tert-Butyl 1-[2-oxo-2-(sodiumoxy)ethyl]piperidine-4-carboxylate (Preparation of (C)

[1068]

[1069] To a solution of B (200.00 mg, 0.737 mmol, 1.00 equiv) in EtOH (5.00 mL) and water (1.00 mL) was added sodium hydroxide (32.43 mg, 0.811 mmol, 1.1 equiv). The resulting solution was stirred at 60 °C overnight. The resulting mixture was concentrated in vacuo to give 180 mg (92.06%) of crude C as a yellow solid. LCMS (ESI) m / z: [M+H] + = 244.

[1070] Step 3: 1-([[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazol-5-yl)phenyl] methyl]aminocarbonyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]aminocarbonyl]methyl)piperidine-4-carboxylate tert-Butyl (D)

[1071]

[1072] To a solution of C (54.25 mg, 0.223 mmol, 1.20 equiv), (2S,4R)-1-[(2S)-2-amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (80.00 mg, 0.186 mmol, 1.00 equiv) and DIPEA (72.04 mg, 0.557 mmol, 3.00 equiv) in DMF (3.00 mL) was added HATU (105.97 mg, 0.279 mmol, 1.50 equiv). The resulting solution was stirred overnight at room temperature. The mixture was concentrated and the residue was purified on a C18 column with ACN:water (7:3). This gave 40 mg (32.82%) of D as a yellow oil. LCMS (ESI) m / z: [M+H] + = 656.

[1073] Step 4: Preparation of 1-([[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazol-5-yl)phenyl] methyl]aminocarbonyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]aminocarbonyl]methyl)piperidine-4- carboxylic Acid (I-80)

[1074]

[1075] To a solution of D (40.00 mg, 0.061 mmol, 1.00 equiv) in DCM (3 mL) was added TFA (0.30 mL, 4.039 mmol, 66.22 equiv). The mixture was stirred overnight at room temperature and then concentrated in vacuo. This gave 120 mg of crude I-80 as a yellow oil. LCMS (ESI) m / z: [M+H] + = 600.

[1076] Preparation of 4-(5-alkylpent-1-yn-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione trifluoroacetate (I-81)

[1077]

[1078] Step 1: Preparation of 2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl Trifluoromethanesulfonate (B) of

[1079]

[1080] To a stirred solution of 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione (A, 2.00 g, 7.293 mmol, 1.00 equiv) in DCM (21.00 mL) was added TEA (2.28 mL, 16.403 mmol, 2.25 equiv) and pyridine (2.28 mL, 28.326 mmol, 3.88 equiv). The reaction mixture was cooled to 0 °C and then trifluoromethanesulfonic anhydride (3.09 g, 10.940 mmol, 1.50 equiv) was added dropwise. The mixture was warmed to room temperature and stirred for 2 h. The resulting mixture was diluted with water (100 mL) and extracted with DCM (3 x 200 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 and concentrated under reduced pressure. The residue was diluted with DCM (50 mL) and stirred at room temperature for 0.5 h. The resulting mixture was filtered and the cake was washed with DCM (10 mL) to give B (1.92 g, 58.32%) as an off-white solid. LCMS (ESI) m / z [M+H] + = 407.

[1081] Step 2: Preparation of tert-Butyl N-[5-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl]pent-4-yn-1- yl]carbamate (C)

[1082]

[1083] To a stirred solution of B (1.60 g, 3.938 mmol, 1.00 equiv) and tert-butyl N-(pent-4-yn-1-yl)carbamate (2.02 g, 11.027 mmol, 2.80 equiv) in THF (40 mL) was added CuI (75.00 mg, 0.394 mmol, 0.10 equiv), DIEA (6.86 mL, 53.074 mmol, 10.00 equiv) and Pd(PPh 3 ) 2 Cl 2 (276.41 mg, 0.394 mmol, 0.10 equiv). The resulting mixture was stirred at 70 °C for 1 h. The reaction mixture was filtered and the cake was washed with EtOAc (3×10 mL). The filtrate was concentrated under reduced pressure and the residue was taken up in water (200 mL) and extracted with EtOAc (2×150 mL). The combined organic layers were washed with brine (200 mL) and dried over anhydrous Na 2 SO 4Dry and concentrate under reduced pressure. Purify the residue by silica gel column chromatography, eluting with petroleum ether / EtOAc (2:1) to give C (1.12 g, 64.72%) as an off-white solid. LCMS (ESI) m / z: [M+H] + = 440.

[1084] Step 3: Preparation of 4-(5-Aminopent-1-yn-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione Trifluoroacetate (I-81)

[1085]

[1086] To a stirred solution of C (1.10 g, 2.503 mmol, 1 equiv) in dichloromethane (6 mL) at room temperature was added TFA (3.00 mL). The resulting mixture was stirred at room temperature for 1 h and then concentrated under reduced pressure. Purify the residue by silica gel flash column chromatography to give I-81 (890 mg, 91.2%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 11.15 (s, 1H), 7.97 – 7.73 (m, 6H), 5.14 (dd, 1H), 3.09 – 2.97 (m, 2H), 2.96 – 2.84 (m, 1H), 2.67 (t, 3H), 2.62 – 2.54 (m, 1H), 2.12 – 2.02 (m, 1H), 1.82 – 1.94 (m, 2H). LCMS (ESI) m / z: [M+H] + = 340.12.

[1087] Preparation of 5-[2-[4-(aminomethyl)piperidin-1-yl]ethoxy]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (I-82)

[1088]

[1089] Step 1: Preparation of 5-(2-Bromoethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (B) of

[1090]

[1091] To a solution of 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindole-1,3-dione (A, 300 mg, 1.09 mmol, 1.00 equiv), KHCO 3(164 mg, 1.64 mmol, 1.50 equivalents) and allyl bromide (198 mg, 1.64 mmol, 1.50 equivalents) were dissolved in DMF (15.00 mL). Then KI (18 mg, 0.11 mmol, 0.10 equivalent) was added to the mixture. The reaction mixture was stirred at 60 °C for 18 h. After cooling to room temperature, 20 mL of water was added, and then the mixture was extracted with EtOAc (3 × 20 mL). The combined organic layers were washed with water (20 mL) and brine (20 mL) and dried over sodium sulfate. After removing the solvent, the residue was purified by flash column chromatography on silica gel, eluting with EtOAc (50%, v / v) in petroleum ether to give B (200 mg, 47.9%) as an off-white solid. LCMS (ESI) m / z: [M+H] + = 315.

[1092] Step 2: Preparation of 2-[[2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindole-5-yl]oxy]acetaldehyde (C) of

[1093]

[1094] To a mixture of t-BuOH (5.00 mL) / water (5.00 mL) / THF (1.00 mL) was added B (250 mg, 0.80 mmol, 1.00 equivalent), NMO (99 mg, 0.80 mmol, 1.5 equivalents) and K 2 OsO 4 (15 mg, 0.04 mmol, 0.050 equivalent). The resulting suspension was stirred at room temperature for 24 h and then remained as a clear solution. The reaction was quenched with saturated Na 2 S 2 O 3 (20 mL) and extracted with EtOAc (3 × 20 mL). The combined organic layers were washed with brine and dried over sodium sulfate. After removing the solvent, the residue was dissolved in a mixture of water (5.00 mL) / acetone (5.00 mL) together with NaIO 4 (255.20 mg, 1.19 mmol, 1.50 equivalents). The reaction mixture was stirred at room temperature for 2 h and then the solvent was removed under vacuum. The residue was suspended in 20 mL of EtOAc. After filtration, the remaining liquid was concentrated under vacuum and the residue was purified by preparative TLC (EtOAc / petroleum ether = 1:1, v / v) to give C (120 mg, 47.7%) as an off-white solid. LCMS (ESI) m / z: [M-H] - = 315.

[1095] Step 3: N-[[1-(2-[[2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindole-5-yl]oxy Preparation of tert-butyl [[2-[4-(aminomethyl)piperidin-1-yl]ethyl]piperidin-4-yl]methylcarbamate (D)

[1096]

[1097] To a solution of C (120 mg, 0.379 mmol, 1.00 equiv) and N-(piperidin-4-ylmethyl)carbamic acid tert-butyl ester (81 mg, 0.38 mmol, 1.0 equiv) in DMF (10.00 mL) was added STAB (120 mg, 0.569 mmol, 1.50 equiv) in one portion. The reaction mixture was stirred overnight at room temperature. After concentration, the residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, water in ACN, 10% to 50% gradient over 10 min; detector, UV 254 nm. This gave D (130 mg, 66.7%) as a white solid. LCMS (ESI) m / z: [M+H] + = 515。

[1098] Step 4: Preparation of 5-[2-[4-(aminomethyl)piperidin-1-yl]ethoxy]-2-(2,6-dioxopiperidin-3-yl)iso indole-1,3-dione (I-82)

[1099]

[1100] D (130.00 mg, 1.00 equiv) was suspended in DCM (10.00 mL), and TFA (1.00 mL) was added. The clear solution was stirred at room temperature for 2 h. Then the solvent was removed under reduced pressure, and the residue I-82 was used without further purification. LCMS (ESI) m / z: [M+H] + = 415。

[1101] Preparation of 1-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]ethyl)piperidine-4-carboxylic acid (I-83)

[1102]

[1103] Step 1: Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-(prop-2-en-1-yloxy)isoindole-1,3-dione (B)

[1104]

[1105] To a solution of 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione (A, 5.48 g, 19.983 mmol, 1.00 equiv) and allyl bromide (3.63 g, 29.975 mmol, 1.5 equiv) in DMF (50.00 mL) was added KI (331.72 mg, 1.998 mmol, 0.1 equiv) and KHCO 3 (3.00 g, 29.975 mmol, 1.5 equiv). The resulting mixture was stirred at 65 °C for 12 h, then diluted with water (100 mL) and extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with brine (3 x 60 mL), dried over anhydrous Na 2 SO 4 and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with hexane / EtOAc (1:1), to give B (6.7 g, crude product) as a yellow-green solid; LCMS (ESI) m / z: [M+H] + = 315.

[1106] Step 2: Preparation of 2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]acetaldehyde (C) Step 3: Preparation of tert-butyl 1-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]eth

[1107]

[1108] To a solution of B (3.14 g, 9.991 mmol, 1.00 equiv) in 1,4-dioxane (30.00 mL) was added NaIO 4 (10.68 g, 49.953 mmol, 5.00 equiv), water (3.00 mL) and 2,6-dimethylpyridine (3.21 g, 29.972 mmol, 3 equiv). To this mixture was added K 2 OsO 4 (0.37 g, 0.999 mmol, 0.1 equiv) dihydrate at room temperature and the reaction was stirred at room temperature for 2 h. The reaction was quenched with water and extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with brine (3 × 100 mL), dried over anhydrous Na 2 SO 4 and concentrated under reduced pressure. This gave C (1.83 g, 57.92%) as a light brown solid; LCMS (ESI) m / z: [M+H] + = 317.

[1109] yl)piperidine-4-carboxylate (D) Step 4: Preparation of 1-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]eth

[1110]

[1111] To a solution of C (1.83 g, 5.786 mmol, 1.00 equiv) and tert-butyl piperidine-4-carboxylate (1.07 g, 5.786 mmol, 1.00 equiv) in DMF (35.00 mL) was added NaBH(OAc) 3 (3.68 g, 17.359 mmol, 3.00 equiv). The resulting mixture was stirred at room temperature for 3 h. After workup with water, extraction with DCM and concentration under reduced pressure, the residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN / 0.1% aqueous formic acid, 0% to 100% gradient over 45 min; detector, UV 254 nm. This gave D (1.16 g, 41.29%) as an off-white solid; LCMS (ESI) m / z: [M+H] + = 401.

[1112] yl)piperidine-4-carboxylic acid (I-83) Step 1: Preparation of 4,5-dimethyl-1H-pyrrole-3-carboxylic acid (B)

[1113]

[1114] To a solution of D (1.16 g, 2.389 mmol, 1.00 equiv) in DCM (10.00 mL) was added TFA (10.00 mL, 134.630 mmol, 34.18 equiv). The reaction mixture was stirred at room temperature for 5 h and then concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN / 0.1% aqueous formic acid, 0% to 100% gradient over 45 min; detector, UV 254 nm. This gave I-83 (845 mg, 73.42%) as a white solid; 11H NMR (300 MHz, DMSO-d6) δ 11.11 (s, 1H), 8.15 (d, 1H), 7.84 (d, 1H), 7.47 (d, 1H), 7.37 (dd, 1H), 5.12 (dd, 1H), 4.31 (t, 2H), 3.02–2.85 (m, 3H), 2.79 (t, 2H), 2.66 - 2.60 (m, 1H), 2.59 - 2.54 (m, 1H), 2.29 - 2.12 (m, 3H), 2.15 - 1.99 (m, 1H), 1.87 - 1.75 (m, 2H), 1.66 – 1.47 (m, 2H); LCMS (ESI) m / z: [M+H] + = 430.15。

[1115] Preparation of 4,5-dimethyl-1-(methylsulfonyl)-N-(2-oxo-2-((4-(3-(pyridin-4-yl)phenyl)thiazol-2-yl)amino)ethyl)-1H-pyrrole-3-carboxamide (I-84)

[1116]

[1117] Step 2: Preparation of tert-butyl 4,5-dimethyl-1H-pyrrole-3-carboxylate (C)

[1118]

[1119] To a solution of methyl 4,5-dimethyl-1H-pyrrole-3-carboxylate (A, 500 mg, 3.26 mmol) in MeOH (4 mL) was added a solution of sodium hydroxide (261.12 mg, 6.53 mmol) in water (1 mL). The mixture was stirred at 25 °C for 36 h. 2M hydrochloric acid (10 mL) and water (10 mL) were added, and the reaction mixture was extracted with EtOAc (50 mL × 2). The combined organic layers were washed with brine (20 mL) and dried over Na 2 SO 4 4 and concentrated under reduced pressure to give B (400 mg, crude) as a yellow solid, which was used without further purification. LCMS (ESI) m / z: [M+H] + = 140.1。

[1120] Step 3: Preparation of tert-butyl 4,5-dimethyl-1-(methylsulfonyl)-1H-pyrrole-3-carboxylate (D)

[1121]

[1122] To a solution of B (400 mg, 2.87 mmol) in toluene (10 mL) was added 1,1 - di - tert - butoxy - N,N - dimethyl - methanamine (2.92 g, 14.37 mmol, 3.45 mL). The mixture was stirred at 80 °C for 2 h. Water (20 mL) was added and the reaction mixture was extracted with EtOAc (50 mL×2). The combined organic layers were washed with brine (20 mL), dried over Na 2 SO 4 and concentrated under reduced pressure. The residue was purified by flash silica chromatography (ISCO; 12 g SepaFlash Silica Flash Column, eluent 0 - 80% EtOAc / petroleum ether gradient at 35 mL / min) and concentrated in vacuo to give C (200 mg, 983.32 μm, 34.21% yield) as a red oil. LCMS (ESI) m / z: [M + H] + = 196.2.

[1123] Step 4: Preparation of 4,5-dimethyl-1-(methylsulfonyl)-1H-pyrrole-3-carboxylic acid (I-84)

[1124]

[1125] Under 0 °C and nitrogen, to a solution of C (120 mg, 0.62 mmol) in THF (3 mL) was added KHMDS (1 M, 1.84 mL). After 0.5 h, methanesulfonyl chloride (140.80 mg, 1.23 mmol, 95.14 μL) was added. The mixture was stirred at 25 °C for 2 h. Water (20 mL) was added and the reaction mixture was extracted with EtOAc (50 mL×2). The combined organic layers were washed with brine (20 mL), dried over Na 2 SO 4 and concentrated under reduced pressure. The residue was purified by flash silica chromatography (ISCO; X g SepaFlash Silica Flash Column, eluent 0 - 50% EtOAc / petroleum ether gradient at 60 mL / min) to give D (80 mg, 263.40 µm, 42.86% yield) as a red solid. LCMS (ESI) m / z: [M + H - 56] + = 218.1.

[1126] Step 1: Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-(pent-4-en-1-yloxy)isoindole-1,3-dione

[1127]

[1128] To a solution of D (80 mg, 0.29 mmol) in 1,4-dioxane (2 mL) was added a solution of 4 M hydrochloric acid in 1,4-dioxane (2 mL). The mixture was stirred at 25 °C for 12 h and then concentrated under reduced pressure to give I-84 (50 mg, crude product) as a purple solid. LCMS (ESI) m / z = [M+H] + = 218.2

[1129] Preparation of 5-[4-(4-aminopiperidin-1-yl)butoxy]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (I-85)

[1130]

[1131] (B) Step 2: Preparation of 4-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]butanal

[1132]

[1133] At room temperature under a nitrogen atmosphere, to a stirred solution of 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindole-1,3-dione (A, 500.00 mg, 1.823 mmol, 1.00 equiv) and KHCO 3 (273.81 mg, 2.735 mmol, 1.50 equiv) in DMF (10.00 mL) was added 5-bromo-1-pentene (326.07 mg, 2.188 mmol, 1.20 equiv) and KI (30.27 mg, 0.182 mmol, 0.10 equiv) portionwise. The resulting mixture was stirred under a nitrogen atmosphere at 65 °C overnight and then cooled and extracted with EtOAc (2 × 20 mL). The combined organic layers were washed with water (3 × 10 mL), dried over anhydrous Na 2 SO 4 and concentrated under reduced pressure. The residue was purified by preparative TLC (petroleum ether / EtOAc 1:1) to give B (400 mg, 64.08%) as a white solid. LCMS (ESI) m / z: [M+H] + = 343

[1134] (C) Step 3: Preparation of tert-butyl N-[1-(4-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]

[1135]

[1136] To a stirred solution of 2-(2,6-dioxopiperidin-3-yl)-5-(pent-4-en-1-yloxy)isoindole-1,3-dione (B, 300.00 mg, 0.876 mmol, 1.00 equiv) in THF (1.00 mL) / water (5.00 mL) / t-BuOH (5.00 mL) was added portionwise NMO (153.98 mg, 1.314 mmol, 1.50 equiv) and K 2 OsO 4 (32.29 mg, 0.088 mmol, 0.10 equiv). The resulting mixture was stirred overnight at room temperature under a nitrogen atmosphere. The reactants were then concentrated under reduced pressure. Then NaIO 4 (374.86 mg, 1.753 mmol, 2.00 equiv), water (0.50 mL) and acetone (5.00 mL) were added and the resulting mixture was stirred for a further 2 h at room temperature. After treatment with aqueous sodium thiosulfate and extraction with DCM, the solution was concentrated under reduced pressure to give C (300 mg, 99.43%) as an off-white solid. The crude product was used directly in the next step without further purification. LCMS (ESI) m / z: [M+H] + = 345.

[1137] butyl)piperidin-4-yl]carbamate (D) Step 4: Preparation of 5-[4-(4-aminopiperidin-1-yl)butoxy]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,

[1138]

[1139] To a solution of 4-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]butanal (C, 270.00 mg, 0.784 mmol, 1.00 equiv) in DMF (5.00 mL) was added portionwise STAB (249.29 mg, 1.176 mmol, 1.50 equiv) and tert-butyl N-(piperidin-4-yl)carbamate (157.05 mg, 0.784 mmol, 1.00 equiv). The resulting mixture was stirred overnight at room temperature and then extracted with EtOAc (2 × 20 mL). The combined organic layers were washed with water (3 × 10 mL), dried over anhydrous Na 2 SO 4 and concentrated under reduced pressure. The residue was purified by preparative TLC (DCM / MeOH 10:1) to give D (130 mg, 31.36%) as an off-white solid. LCMS (ESI) m / z: [M+H] + = 529.

[1140] 3-dione (I-85) Step 1: Preparation of tert-butyl 4-[1-[2-[2-(2,6-dioxo-3-piperidinyl)-1,3-dioxo-isoindolin-4-yl]oxy

[1141]

[1142] At room temperature, a solution of tert-butyl N-[1-(4-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]butyl)piperidin-4-yl]carbamate (D, 120.00 mg, 0.227 mmol, 1.00 equiv) and TFA (1.00 mL) in DCM (4.00 mL) was stirred for 2 h. The resulting mixture was concentrated under reduced pressure. This gave I-85 (130 mg, 93.55%) as an off-white solid. LCMS (ESI) m / z: [M+H] + = 429.

[1143] Preparation of tert-butyl 2-[(4-bromo-2-pyridinyl)methyl-amino]acetate (I-86)

[1144]

[1145] To a solution of 4-bromo-2-fluoro-pyridine (A, 28 g, 159.10 mmol) and tert-butyl 2-(methylamino)acetate (34.68 g, 190.92 mmol, HCl) in DMSO (200 mL) was added DIEA (41.13 g, 318.21 mmol, 55.43 mL). The mixture was stirred at 120 °C for 12 h. The reaction mixture was diluted with water (2 L) and extracted with EtOAc (500 mL x 3). The combined organic phases were washed with brine (500 mL x 2), dried over Na 2 SO 4 dried, filtered and concentrated. The residue was purified by flash column chromatography (silica gel, petroleum ether / EtOAc 10:1 to 5:1) to give I-86 (35 g, 104.59 mmol, 65.74%) as a white solid. LCMS (ESI) m / z: [M+H] + = 301.0. 1 H NMR (400 MHz, CDCl 3 ) δ = 7.86 (d, J J = 5.2 Hz, 1H), 6.66 - 6.65 (m, 1H), 6.62 (d, J J = 1.2 Hz, 1H), 4.12 (s, 2H), 3.00 (s, 3H), 1.35 (s, 9H) ppm.

[1146] Preparation of 2-(2,6-dioxo-3-piperidinyl)-4-[2-oxo-2-[4-(4-piperidinyl)-1-piperidinyl]ethoxy]isoindoline-1,3-dione (I-87)

[1147]

[1148] acetyl]-4-piperidinyl]piperidine-1-carboxylate (B) Step 2: Preparation of 2-(2,6-dioxo-3-piperidinyl)-4-[2-oxo-2-[4-(4-piperidinyl)-1-piperidinyl]eth

[1149]

[1150] To a solution of tert-butyl 4-(4-piperidinyl)piperidine-1-carboxylate (A, 730 mg, 2.72 mmol), HOBt (551.28 mg, 4.08 mmol), DIEA (1.76 g, 13.60 mmol, 2.37 mL) and 2-[2-(2,6-dioxo-3-piperidinyl)-1,3-dioxoisoindol-4-yl]oxyacetic acid (903.72 mg, 2.72 mmol) in DMF (20 mL) was added EDCI (782.11 mg, 4.08 mmol). The mixture was then stirred at 25 °C for 8 h. Water (30 mL) was added and the mixture was extracted with EtOAc (45 mL x 3). The combined organic extracts were dried over anhydrous Na 2 SO 4 and concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography (0.1% aqueous formic acid / ACN). The solution was then concentrated to remove ACN and the residue was extracted with EtOAc (30 mL x 3). The combined organic phases were dried over anhydrous Na 2 SO 4 dried, filtered and concentrated under reduced pressure to give B (820 mg, 1.41 mmol, 51.74% yield) as a yellow solid. LCMS (ESI) m / z: [M-100] + = 483.2.

[1151] oxy]isoindoline-1,3-dione (I-87) Step 1: Preparation of 4-(3-bromophenyl)thiazol-2-amine (B)

[1152]

[1153] To a solution of B (750 mg, 1.29 mmol) in DCM (10 mL) at 0 °C was added TFA (3.08 g, 27.01 mmol, 2 mL). The mixture was then stirred at 25 °C for 2 h and then concentrated to give I-87 (700 mg, 1.17 mmol, 91.16% yield, TFA) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 783.2. 1 H NMR (400MHz, DMSO-d 6 ) δ = 8.37 (s, 1H), 7.82 - 7.80 (m, 1H), 7.50 (d, J = 7.2 Hz, 1H),7.37 (d, J = 8.6 Hz, 1H), 5.19 (s, 2H), 4.44 - 4.34 (m, 1H), 4.09 (d, J = 7.2Hz, 1H), 3.90 - 3.79 (m, 2H), 3.03 - 3.02 (m, 4H), 2.71 - 2.58 (m, 2H), 2.49 - 2.48(m, 2H), 2.14 - 2.06 (m, 1H), 2.05 (s, 1H), 1.71 (s, 2H), 1.62 - 1.61 (m, 2H),1.23 - 1.22 (m, 4H), 1.10 (d, J = 6.0 Hz, 2H) ppm。

[1154]

[1155] Preparation of 1-(Methylsulfonyl)-N-(2-oxo-2-((4-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)thiazol-2-yl)amino)ethyl)-1H-pyrrole-3-carboxamide (I-90) and N-(2-((4-(3-bromophenyl)thiazol-2-yl)amino)-2-oxoethyl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide (I-92)

[1156]

[1157] Step 2: Preparation of tert-butyl N-[2-[[4-(3-bromophenyl)thiazol-2-yl]amino]-2-oxo-ethyl]carbamate

[1158]

[1159] To a mixture of 1-(3-bromophenyl)ethanone (A, 473 g, 2.38 mol, 313.25 mL) and thiourea (361.78 g, 4.75 mol) was added I 2 (603.14 g, 2.38 mol, 1 equivalent). The mixture was stirred at 110 °C for 16 h. After cooling, the reaction mixture was triturated with MTBE (5 L) and then filtered to remove any unreacted iodine and acetophenone. The cake was suspended in ice water (4 L) and treated with 25% aqueous ammonia to pH 9 - 10. The suspension was stirred at 25 °C for 15 min, then filtered and washed with water (1 L) to give a wet solid. It was dissolved in EtOAc (4 L) and washed with saturated NaHCO 3 aqueous solution (1 L × 2) and brine (1 L). The EtOAc layer was dried over Na 2 SO 4 , filtered and concentrated under reduced pressure. The residue was stirred with petroleum ether / EtOAc 100:1 (4 L) at 25 °C for 3 h, then the suspension was filtered, the cake was washed with petroleum ether (1 L) and dried in vacuo to give B as a pink solid (450 g, 1.69 mol, 71.20% yield, 95.93% purity). 1 H NMR (400 MHz, DMSO-d 6 ) δ = 7.98 - 7.97 (m, 1H), 7.80 - 7.77 (m,1H), 7.43 - 7.42 (m, 1H), 7.34 - 7.30 (m, 1H), 7.15 (s, 1H), 7.10 (s, 2H). LCMS(ESI) m / z: 79 Br M+H + = 254.9.

[1160] (C) ​

[1161]

[1162] To a solution of 2-(tert-butoxycarbonylamino)acetic acid (82.40 g, 470.34 mmol), HATU (178.84 g, 470.34 mmol), and DIEA (151.97 g, 1.18 mol, 204.81 mL) in DCM (1 L) was added B (100.00 g, 391.95 mmol), and the mixture was stirred at 30 °C for 16 h. The reaction mixture was washed with saturated aqueous citric acid (500 mL x 4) and brine (500 mL), dried over Na 2 SO 4 and concentrated under reduced pressure. The residue was triturated with MeOH (200.0 mL), and the resulting liquid was concentrated under reduced pressure to afford C (100 g, 241.89 mmol, 61.71% yield) as a white solid. 1 1H NMR (400 MHz, DMSO-d 6 ) δ = 12.29 (s, 1H), 8.09 - 8.09 (m, 1H), 7.89 (d, J = 7.6 Hz, 1H), 7.76 (s, 1H), 7.52 - 7.49 (m, 1H), 7.41 - 7.37 (m, 1H), 7.16 - 7.13 (m, 1H), 3.87 - 3.81 (m, 2H), 1.39 (s, 9H) ppm. LCMS (ESI) m / z: 81 [BrM+H] + = 413.8.

[1163] Step 3: Preparation of 2-Amino-N-(4-(3-bromophenyl)thiazol-2-yl)acetamide Hydrochloride (D)

[1164]

[1165] A mixture of C (10 g, 24.25 mmol) and 4 M HCl in 1,4-dioxane (100 mL) was stirred at 30 °C for 2 h. The reaction mixture was concentrated in vacuo to afford D (8.4 g, crude, HCl) as a white solid, which was used without further purification. LCMS (ESI) m / z: [M+H] + = 313.8.

[1166] Step 4: Preparation of N-(2-((4-(3-bromophenyl)thiazol-2-yl)amino)-2-oxoethyl)-1-(methanesulfonyl)- 1H-pyrrole-3-carboxamide (I-92)

[1167]

[1168] To a solution of D (8.4 g, 24.09 mmol) and 1-(Methylsulfonyl)-1H-pyrrole-3-carboxylic acid (5.47 g, 28.91 mmol) in DCM (100 mL) was added HATU (10.99 g, 28.91 mmol) and DIEA (18.68 g, 144.56 mmol, 25.18 mL). The mixture was stirred at 20 °C for 16 h. The resulting suspension was filtered and triturated with MTBE (50 mL x 2) to give a filter cake, which was dried under vacuum to give I-92 (10 g, 20.58 mmol, 85.43% yield) as a white solid, which was used without further purification. 1 1H NMR (400 MHz, DMSO-d6) δ = 12.40 - 12.35 (m, 1H), 8.69 - 8.66 (m, 1H), 8.11 - 8.10 (m, 1H), 7.92 - 7.90 (m, 1H), 7.85 - 7.84 (m, 1H), 7.78 (s, 1H), 7.53 - 7.51 (m, 1H), 7.42 - 7.38 (m, 1H), 7.32 - 7.30 (m, 1H), 6.78 - 6.77 (m, 1H), 4.14 (d, J J = 6.0 Hz, 2H), 3.57 (s, 3H). LCMS (ESI) m / z: [M+H] + = 484.8.

[1169] Step 5: Preparation of 1-(Methanesulfonyl)-N-(2-oxo-2-((4-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan- 2-yl)phenyl)thiazol-2-yl)amino)ethyl)-1H-pyrrole-3-carboxamide (I-90)

[1170]

[1171] To a solution of I-92 (1.5 g, 3.10 mmol) and 4,4,5,5-Tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (1.18 g, 4.65 mmol) in 1,4-Dioxane (15 mL) was added Pd(dppf)Cl 2 2 (227.07 mg, 0.310 mmol) and KOAc (913.69 mg, 9.31 mmol). The mixture was stirred at 80 °C for 2 h, then poured into water (100 mL) and extracted with EtOAc (100 mL x 3). The combined organic layers were washed with brine (50 mL) and dried over Na 2 2SO 4Dry and concentrate under reduced pressure. The residue was suspended in 1:1 petroleum ether / EtOAc (20 mL), filtered, and the filtrate was concentrated under vacuum to give I-90 as a brown solid (6.5 g, 12.00 mmol, 96.71% yield). 1 H NMR (400 MHz, DMSO-d6) δ= 12.45 (s, 1H), 8.67 (t, J J = 6.0 Hz, 1H), 8.29 (s, 1H), 8.01 (br d, J J = 7.6 Hz, 1H), 7.84 (s,1H), 7.65(s, 1H), 7.62 (d, J J = 7.2 Hz, 1H), 7.44 (t, J J = 7.6 Hz, 1H), 7.31 (t, J J = 2.8Hz, 1H), 6.78 (d, J J = 1.6 Hz, 1H), 4.14 (d, J J = 6.0 Hz, 2H), 3.57 (s, 3H),1.31 (s, 12H). LCMS (ESI) m / z: [M+H] + = 531.2。

[1172] Preparation of tert-butyl 1-(4-(3-(2-(2-(1-(methylsulfonyl)-1H-pyrrole-3-carboxamido)acetamido)thiazol-4-yl)phenyl)pyridin-2-yl)cyclopropanecarboxylate (I-91)

[1173]

[1174] Step 1: Preparation of tert-Butyl 2-(4-bromopyridin-2-yl)acetate (B)

[1175]

[1176] At -78 °C, LDA (2 M in THF, 100 mL, 200 mmol) was added dropwise to a solution of 4-bromo-2-methyl-pyridine (A, 10 g, 36.9 mmol) in THF (150 mL). After addition, the mixture was stirred at -78 °C for 1 h. Then Boc in THF was added dropwise at -78 °C 2O (14.0 g, 64.2 mmol). The resulting mixture was stirred overnight at 25 °C, then quenched with water (300 mL) and extracted with EtOAc (200 mL × 3). The combined organic layers were washed with brine (100 mL × 3), dried over Na 2 SO 4 and concentrated under reduced pressure to give B (17.2 g) as a black crude oil. LCMS (ESI) m / z [M+H] + = 272.

[1177] Step 2: Preparation of tert-Butyl 1-(4-bromopyridin-2-yl)cyclopropanecarboxylate (C)

[1178]

[1179] To a solution of B (5.0 g, 18.45 mmol) in DMF (50 mL) at 0 °C was added NaH (1.5 g, 37.5 mmol, 60%, in mineral oil). The mixture was stirred at this temperature for 1 h, then 1,2-dibromoethane (6.8 g, 37.5 mmol) was added dropwise at 0 °C. The resulting mixture was stirred at 25 °C for 1 h. The reaction mixture was quenched by the addition of water (200 mL), followed by extraction with EtOAc (100 mL × 3). The combined organic layers were washed with brine (100 mL × 3), dried over Na 2 SO 4 and filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with petroleum ether / EtOAc (2:1) to give C (3.7 g, 12.4 mmol, 67.5% yield) as a yellow oil. LCMS (ESI) m / z [M+H] + = 298.

[1180] Step 3: 1-(4-(3-(2-(2-(1-(Methanesulfonyl)-1H-pyrrole-3-carboxamido)acetamido)thiazol- 4-yl)phenyl)pyridin-2-yl)cyclopropanecarboxylate (D)

[1181]

[1182] C (618.4 mg, 2.07 mmol), I-90 (1.0 g, 1.88 mmol), K 3 PO 4A mixture of (800.4 mg, 3.77 mmol) and bis(tert-butyl)(cyclopentyl)phosphine palladium(II) dichloride (24.57 mg, 0.03 mmol) in 1,4-dioxane (20 mL) / water (2 mL) was degassed and purged with nitrogen three times. The reaction mixture was stirred at 80 °C for 12 h, then quenched with water (20 mL), and subsequently extracted with EtOAc (100 mL × 3). The combined organic layers were washed with saturated aqueous NaCl solution (100 mL × 3), dried over Na 2 SO 4 and concentrated under reduced pressure. The residue was purified by reverse-phase chromatography (ACN / 0.1% formic acid) and lyophilized to afford D (830 mg, 1.33 mmol, 70.81%) as a yellow solid. LCMS (ESI) m / z [M+H] + = 622.

[1183] Step 4: Preparation of 1-(4-(3-(2-(2-(1-(Methanesulfonyl)-1H-pyrrole-3-carboxamido)acetamido)thiazol- 4-yl)phenyl)pyridin-2-yl)cyclopropanecarboxylic Acid (I-91)

[1184]

[1185] A mixture of D (830 mg, 1.33 mmol) and TFA (2 mL) in DCM (10 mL) was stirred at 25 °C for 1 h. The mixture was concentrated and the residue was used without further purification (I-91, 1.2 g), as a yellow solid. LCMS (ESI) m / z [M+H] + = 566.

[1186] Compound 11-N-[2-[[4-[3-[2-[1-[2-[2-[2-[[2-(2,6-dioxo-3-piperidinyl)-1,3-dioxo-isoindoline-4-yl]amino]ethoxy]ethoxy]ethylcarbamoyl]cyclopropyl]-4-pyridinyl]phenyl] thiazol-2-yl]amino]-2-oxo-ethyl]-1-methanesulfonylpyrrole-3-carboxamide Preparation Step 1: Preparation of tert-Butyl 3-bromobenzylcarbamate (B)

[1187]

[1188] A solution of 1-[4-[3-[2-[[2-[(1-Methanesulfonylpyrrole-3-carbonyl)amino]acetyl]amino]thiazol-4-yl]phenyl]-2-pyridyl]cyclopropanecarboxylic acid (I-91, 60 mg, 106.08 µm), HATU (121.00 mg, 318.23 µm), and DIEA (68.55 mg, 530.39 µm, 92.38 µL) in DMF (1 mL) was stirred at 30 °C for 1 hour. To this mixture was added 4-[2-[2-(2-Aminoethoxy)ethoxy]ethylamino]-2-(2,6-dioxo-3-piperidinyl)isoindoline-1,3-dione (I-9, 55.00 mg, 0.11 mmol), and then the mixture was stirred at 30 °C for 12 hours. The mixture was triturated with water (3 mL) and filtered. The filter cake was purified by reverse-phase HPLC (column: Phenomenex Synergi C18 150x30mmx4µm; mobile phase: [0.225% aqueous formic acid - ACN]; 30% - 60%) to give Compound 11 as a yellow solid (21.27 mg, 21.10 µm, 19.89%, yield, 98.991%). 1 H NMR (400 MHz, chloroform-d) δ = 9.36 (d, J = 1.6 Hz, 1H), 8.59 (d, J = 5.2 Hz, 1H), 8.13 (s, 1H), 7.80 - 7.75 (m, 2H), 7.61 (s, 2H), 7.58 - 7.54 (m, 1H), 7.50 (d, J = 8.0 Hz, 1H), 7.43 - 7.35 (m, 2H), 7.19 (s, 1H), 7.13 (m, 1H), 7.08 (d, J = 7.2 Hz, 1H), 6.85 (m, 1H), 6.76 (d, J = 8.8 Hz, 1H), 6.70 (m, 1H), 6.38 (m, 1H), 4.99 - 4.95 (m, 1H), 4.39 (m, 2H), 3.61 - 3.46 (m, 10H), 3.33 - 3.28 (m, 2H), 3.25 - 3.25 (m, 1H), 3.23 (s, 2H), 2.89 - 2.73 (m, 3H), 2.15 - 2.10 (m, 1H), 1.75 - 1.70 (m, 2H), 1.35 - 1.30 (m, 1H), 1.25 (m, 1H) ppm.

[1189]

[1190]

[1191]

[1192]

[1193]

[1194]

[1195]

[1196]

[1197]

[1198]

[1199]

[1200]

[1201]

[1202]

[1203]

[1204] Preparation of N-(2-((4-(3'-(aminomethyl)-[1,1'-biphenyl]-3-yl)thiazol-2-yl)amino)-2-oxoethyl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide (I-93)

[1205]

[1206] Step 2: Preparation of tert-Butyl 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzylcarbamate

[1207]

[1208] To a solution of (3-bromophenyl)methanamine (A, 3.7 g, 19.89 mmol) in THF (20 mL) was added NaHCO 3 (3.34 g, 39.77 mmol, 1.55 mL) and Boc 2O (4.77 g, 21.88 mmol, 5.03 mL). The mixture was stirred at 30 °C for 16 h, filtered, and concentrated to give B (5.6 g, 16.67 mmol, 83.85%) as a white solid, which was used without further purification. LCMS (ESI) m / z [M+H-56] + = 231.9.

[1209] (C) Step 3: Preparation of tert-Butyl ((3'-(2-(2-(1-(Methanesulfonyl)-1H-pyrrole-3-carboxamido)acetamido)thiazol-4-

[1210]

[1211] To a solution of B (5.6 g, 16.67 mmol) and bis(pinacolato)diboron (5.08 g, 20.01 mmol) in 1,4-dioxane (60 mL) was added cyclopentyl(diphenyl)phosphine dichloropalladium (II), DCM adduct (1.36 g, 1.67 mmol) and KOAc (4.91 g, 50.02 mmol). The mixture was stirred at 80 °C for 2 h and then poured into water (100 mL). The mixture was extracted with EtOAc (100 mL x 3), and the combined organic layers were washed with brine (200 mL), dried over Na 2 SO 4 4, filtered, and concentrated. The residue was purified by column chromatography (SiO 2 , petroleum ether / EtOAc 1:0 - 10:1) to give C (5.5 g, 16.51 mmol, 98.98%) as a white solid. 1 1H NMR (400 MHz, DMSO-d6) δ 7.58 (s, 1H), 7.53 (d, J = 6.8 Hz, 1H), 7.35 - 7.32 (m, 3H), 4.13 (d, J = 6.0 Hz, 2H), 1.39 (s, 9H), 1.29 (s, 12H).

[1212] yl)-[1,1'-biphenyl]-3-yl)methyl)carbamate (D) Step 4: Preparation of N-(2-((4-(3'-(Aminomethyl)-[1,1'-biphenyl]-3-yl)thiazol-2-yl)amino)-2-

[1213]

[1214] To a solution of N-[2-[[4-(3-bromophenyl)thiazol-2-yl]amino]-2-oxo-ethyl]-1-methanesulfonyl-pyrrole-3-carboxamide (I-92, 500 mg, 1.03 mmol) and C (413.64 mg, 1.24 mmol) in 1,4-dioxane (5 mL) / water (0.5 mL) was added Pd(dppf)Cl 2 (75.69 mg, 103.44 μmol) and K 2 CO 3 (428.89 mg, 3.10 mmol). The mixture was stirred at 80 °C for 2 h, then concentrated, and the residue was purified by reverse phase flash HPLC (0.1% aqueous formic acid / ACN) and lyophilized to give D (600 mg) as a yellow solid. LCMS (ESI) m / z [M+H] + = 610.2.

[1215] oxoethyl)-1-(methanesulfonyl)-1H-pyrrole-3-carboxamide (I-93) Step 1: Preparation of 4-[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]thiazol-2-amine (B)

[1216]

[1217] A mixture of D (400 mg, 656.04 μmol) and 4M HCl in 1,4-dioxane (5 mL) was stirred at 30 °C for 2 h. The reaction mixture was concentrated and the residue was triturated with MTBE (2 mL), filtered to give a yellow solid, and purified by preparative HPLC twice (column: Phenomenex Synergi C18 150x25x10 μm; mobile phase: [water (0.05% HCl)-ACN]; B%: 18%-38%, 10 min), then (column: Shim-pack C18 150x25x10 μm; mobile phase: [0.225% aqueous formic acid / ACN]; B%: 10%-40%, 10 min) to give I-93 (68.49 mg, 134.40 μmol, 56.45%) as a white solid. LCMS (ESI) m / z [M+H] + = 510.2. 1HNMR (400 MHz, DMSO-d6) δ 8.69 - 8.67 (m, 1H), 8.33 (s, 1H), 8.20 (s, 1H), 7.90 (d, J = 8.0 Hz, 1H), 7.85 - 7.83 (m, 1H), 7.75 (s, 2H), 7.64 - 7.60 (m, 2H), 7.55 - 7.52 (m, 1H), 7.49 - 7.45 (m, 1H), 7.40 - 7.38 (m, 1H), 7.32 - 7.30 (m, 1H), 6.78 (dd, J = 1.6, 3.2 Hz, 1H), 4.15 (d, J = 5.6 Hz, 2H), 3.92 (s, 2H), 3.57 (s, 3H).

[1218] Preparation of Compound 1 – N-(2-((4-(3'-((S)-14-((2S,4R)-4-Hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carbonyl)-15,15-dimethyl-3,12-dioxo-6,9-dioxa-2,13-diazacyclohexadecyl)-[1,1'-biphenyl]-3-yl)thiazol-2-yl)amino)-2-oxoethyl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide

[1219]

[1220] To a solution of N-[2-[[4-[3-[3-(aminomethyl)phenyl]phenyl]thiazol-2-yl]amino]-2-oxo-ethyl]-1-methylsulfonyl-pyrrole-3-carboxamide (I-93, 50 mg, 0.098 mmol) and 3-[2-[3-[[1-[(2S,4R)-4-hydroxy-2-[[4-(4-methylthiazol-5-yl)phenyl]methylcarbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-3-oxo-propoxy]ethoxy]propanoic acid (I-52, 66.78 mg, 0.107 mmol) in DCM (2 mL) was added EDCI (22.57 mg, 117.74 µm), HOBt (15.91 mg, 0.117 mmol) and DIEA (12.68 mg, 0.098 mmol, 17.09 µL). The mixture was stirred at 30 °C for 2 h and then concentrated. The residue was purified by preparative HPLC (column: Phenomenex Synergi C18 150 x 25 x 10 µm; mobile phase: [0.225% aqueous formic acid / ACN]; B%: 33% - 63%, 10 min) to give compound 1 as a white solid (44.16 mg, 40.54% yield). LCMS (ESI) m / z: [M+Na] + = 1132.3. 1 H NMR (400 MHz, methanol-d4) δ = 8.84 (s,1H), 8.18 - 8.12 (m, 1H), 7.89 - 7.81 (m, 2H), 7.60 (s, 1H), 7.55 (br d, J = 6.6Hz, 2H), 7.49 - 7.34 (m, 7H), 7.32 - 7.25 (m, 2H), 6.80 (dd, J= 1.6, 3.4 Hz, 1H), 4.63 (s, 1H), 4.59 - 4.44 (m, 6H), 4.35 - 4.25 (m, 3H), 3.91 - 3.84 (m, 1H), 3.80 - 3.70 (m, 3H), 3.62 - 3.49 (m, 4H), 3.48 - 3.39 (m, 2H), 3.37 (s, 3H), 2.50 - 2.48 (m, 2H), 2.46 - 2.42 (m, 3H), 2.41 - 2.29 (m, 2H), 2.24 - 2.14 (m, 1H), 2.10 - 2.01 (m, 1H), 1.05 - 0.93 (m, 9H) ppm。

[1221]

[1222]

[1223]

[1224]

[1225]

[1226]

[1227]

[1228]

[1229]

[1230]

[1231]

[1232]

[1233] Preparation of 2-(methyl(4-(3-(2-(2-(1-(methanesulfonyl)-1H-pyrrole-3-carboxamido)acetamido)thiazol-4-yl)phenyl)pyridin-2-yl)amino)acetic acid (I-94)

[1234]

[1235] At 30 °C under nitrogen, to a solution of 1-(methylsulfonyl)-N-[2-oxo-2-[[4-[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]thiazol-2-yl]amino]ethyl]pyrrole-3-carboxamide (I-90, 100 mg, 0.188 mmol) and 2-[(4-bromo-2-pyridinyl)methylamino]-N,N-dimethylacetamide (from I-86, 51.31 mg, 0.188 mmol) in 1,4-dioxane (2 mL) / water (0.5 mL) was added bis(tert-butyl)(cyclopentyl)phosphine palladium(II) dichloride (12.29 mg, 0.018 mmol) and K 3 PO 4 (120.06 mg, 0.57 mmol). The reaction mixture was stirred at 75 °C for 2 h and then concentrated under reduced pressure. The residue was purified by reverse phase HPLC (0.1% aqueous formic acid / ACN) (column: Phenomenex Synergi C18 150 x 25 x 10 µm) to give I-94 (10 mg, 8.63%) as a white solid. LCMS (ESI) m / z: [M+H] + = 569.3. 1 H NMR (400 MHz, DMSO-d6) δ = 12.40 (br s, 1H), 8.69 - 8.66 (m, 1H), 8.21 (s, 1H), 8.14 (d, J = 5.2 Hz, 1H), 7.97 (d, J = 7.6 Hz, 1H), 7.87 - 7.78 (m, 2H), 7.69 (d, J = 7.6 Hz, 1H), 7.61 - 7.52 (m, 1H), 7.32 - 7.30 (m, 1H), 6.96 - 6.85 (m, 2H), 6.77 (br s, 1H), 4.31 (s, 2H), 4.15 (d, J = 5.6 Hz, 2H), 3.56 (s, 3H), 3.13 (s, 3H) ppm。

[1236] Preparation of 9-N-(2-((4-(3-(2-((2-((2-(2-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethyl)amino)-2-oxoethyl)(methyl)amino)pyridin-4-yl)phenyl)thiazol-2-yl)amino)-2-oxoethyl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide

[1237]

[1238] To a solution of 2-[methyl-[4-[3-[2-[[2-[(1-methylsulfonylpyrrole-3-carbonyl)amino]acetyl]amino]thiazol-4-yl]phenyl]-2-pyridinyl]amino]acetic acid (I-94, 50 mg, 0.082 mmol), DIEA (42.72 mg, 0.33 mmol, 57.57 μL), EDCI (15.84 mg, 0.082 mmol) and HOBt (11.16 mg, 0.082 mmol) in DMF (1 mL) was added 4-[2-[2-(2-aminoethoxy)ethoxy]ethylamino]-2-(2,6-dioxo-3-piperidinyl)isoindoline-1,3-dione (I-9, 42.84 mg, 0.082 mmol). The mixture was stirred at 25 °C for 2 h, then poured into water (40 mL), filtered, triturated with water (10 mL x 2) and dried in vacuo to give compound 9 (27.03 mg, 32.88%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 955.3. 1 H NMR (400 MHz, DMSO-d 6) δ = 12.40 (s, 1H), 11.08 (s, 1H), 8.69 - 8.66 (m, 1H), 8.20 - 8.13 (m, 2H), 7.97 (d, J = 8.0 Hz, 1H), 7.87 - 7.84 (m, 2H), 7.78 (s, 1H), 7.67 (d, J = 8.0 Hz, 1H), 7.59 - 7.53 (m, 2H), 7.32 - 7.31 (m, 1H), 7.11 (d, J = 8.8 Hz, 1H), 7.03 (d, J = 6.8 Hz, 1H), 6.92 (d, J = 5.2 Hz, 1H), 6.86 (s, 1H), 6.78 - 6.77 (m, 1H), 6.60 - 6.57 (m, 1H), 5.07 - 5.02 (m, 1H), 4.21 - 4.14 (m, 4H), 3.60 - 3.58 (m, 5H), 3.51 - 3.50 (m, 4H), 3.45 - 3.40 (m, 4H), 3.25 - 3.22 (m, 2H), 3.12 (s, 3H), 2.88 - 2.86 (m, 1H), 2.59 (s, 2H), 2.06 - 1.98 (m, 1H) ppm。

[1239]

[1240]

[1241]

[1242]

[1243]

[1244]

[1245]

[1246]

[1247]

[1248] Compound 123 - N - [2 - [[4 - [3 - [2 - [[2 - [4 - [1 - [2 - [2 - (2,6 - dioxo - 3 - piperidinyl)-1,3 - dioxoisoindolin - 4 - yl]oxyacetyl]-4 - piperidinyl]-1 - piperidinyl]-2 - oxo - ethyl]-methyl - amino]-4 - pyridinyl]phenyl]thiazol - 2 - yl]amino]-2 - oxo - ethyl]-4,5 - dimethyl - 1 - mesyl - pyrrole - 3 - carboxamide

[1249]

[1250] Step 2: Preparation of tert-Butyl Intermediate 52-[[4-[3-(2-aminothiazol-4-yl)phenyl]-2-pyridinyl]-methyl-amino]acetate (C) Step 3: 2-((4-(3-(2-(2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)acetamido)thiazol-

[1251]

[1252] To a solution of 4-(3 - bromophenyl)thiazol - 2 - amine (A, 20 g, 78.39 mmol) and bis(pinacolato)diboron (29.86 g, 117.59 mmol) in 1,4 - dioxane (200 mL) was added Pd(dppf)Cl2 (5.74 g, 7.84 mmol) and KOAc (23.08 g, 235.17 mmol). The mixture was stirred at 70 °C for 2 h, then diluted with water (2 L), extracted with EtOAc (500 mL x 3), the organic layers were combined, dried over Na 2 SO 4 and concentrated. The residue was purified by column chromatography (SiO 2 , petroleum ether / EtOAc 10:1 to 5:1) to give B (21 g, 78.98%) as a brown solid. LCMS (ESI) m / z: [M + H] + = 303.1. 1 H NMR (400 MHz, CDCl 3 ) δ = 8.20 (s,1H), 7.87 (d, J = 7.6 Hz, 1H), 7.73 (d, J = 7.2 Hz, 1H), 7.40 (d, J = 7.6 Hz,1H), 6.76 (s, 1H), 5.64 (s, 2H), 1.38 (s, 12H) ppm。

[1253] 4-yl)phenyl)thiazol-2-yl)amino)-2-oxoethyl)-1-(methanesulfonyl)-1H-pyrrole-3-carboxamide (I-94) Preparation Step 4: Preparation of 2-((4-(3-(2-(2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)acetamido)thiazol-

[1254]

[1255] B (13.2 g, 43.83 mmol), I-86 (14.57 g, 48.21 mmol), K 3 PO 4 (27.91 g, 131.48 mmol), and bis(tert-butyl)(cyclopentyl)phosphine palladium(II) dichloride (2.86 g, 4.38 mmol) in a mixture of 1,4-dioxane (150 mL) / water (40 mL) were degassed and purged with nitrogen three times. The mixture was stirred at 80 °C for 2 h, then diluted with water (1 L) and extracted with EtOAc (300 mL x 3). The combined organic phases were dried over Na 2 SO 4 , filtered, and concentrated. The residue was purified by column chromatography (SiO 2 , petroleum ether / EtOAc 10:1 to 1:1). Subsequently, the solid material was triturated with MeOH (200 mL) to give C (15 g, 35.94 mmol, 63.13% 1) as a brown solid. LCMS (ESI) m / z: [M+H] + = 397.2. 1 H NMR (400 MHz, DMSO-d 6 ) δ = 8.15 - 8.08 (m, 2H), 7.86 (d, J = 7.6 Hz, 1H), 7.61 (d, J = 8.0 Hz, 1H), 7.50 - 7.46 (m, 1H), 7.17 (s, 1H), 7.09 (s, 2H), 6.92 - 6.90 (m, 1H), 6.86 (s, 1H), 4.28 (s, 2H), 3.12 (s, 3H), 1.37 (s, 9H) ppm.

[1256] 4-yl)phenyl)thiazol-2-yl)amino)-2-oxoethyl)-1-(methanesulfonyl)-1H-pyrrole-3-carboxamide (I-94) Preparation of tert-Butyl (4-(4-(3-(2-((2-Aminoacetyl)amino)thiazol-4-yl)phenyl)pyridin-2-yl)(methyl)amino)acetate (D)

[1257]

[1258] EDCI (9.67 g, 50.44 mmol) was added to a solution of C (12.00 g, 40.35 mmol) in pyridine (60 mL). The mixture was stirred at 25 °C for 0.5 h. Then tert-butyl 2-[[4-[3-(2-aminothiazol-4-yl)phenyl]-2-pyridinyl]methylamino]acetate (4 g, 10.09 mmol) was added and the reaction mixture was stirred at 25 °C for an additional 12 h. The reaction mixture was slowly poured into water (600 mL) and filtered. The filter cake was purified by column chromatography (SiO 2 , DCM / EtOAc 1:0 to 5:1) to give D (5.2 g, 72.46% yield) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 676.2. 1 H NMR (400 MHz, DMSO-d 6 ) δ = 12.37 (s, 1H), 8.22 (s, 1H), 8.15 (d, J = 4.8 Hz, 1H), 7.98 (d, J = 7.2 Hz, 1H), 7.91 (d, J = 7.6 Hz, 2H), 7.81 (s,1H), 7.77 - 7.68 (m, 4H), 7.60 - 7.54 (m, 1H), 7.43 (d, J = 7.6 Hz, 2H), 7.38 - 7.33 (m, 2H), 6.95 - 6.90 (m, 1H), 6.91 (s, 1H), 4.36 - 4.32 (m, 2H), 4.30 (s,2H), 4.27 (d, J = 6.8 Hz, 1H), 3.97 (d, J = 6.0 Hz, 2H), 3.14 (s, 3H), 1.39(s, 9H) ppm.

[1259] Step 4: Preparation of tert-Butyl 2-((4-(3-(2-((2-Aminoacetyl)amino)thiazol-4-yl)phenyl)pyridin-2-yl)methylamino)acetate (E) Step 5: Preparation of tert-Butyl 2-((4-(3-(2-((2-((4,5-Dimethyl-1-(methylsulfonyl)pyrrol-3-carbonyl)amino)acetyl)amino)thiazol-4-yl)phenyl)pyridin-2-yl)methylamino)acetate (F)

[1260]

[1261] A mixture of D (5.1 g, 7.55 mmol) and dimethylamine in THF (2.0 M, 120 mL) was stirred at 25 °C for 1 h. The reaction mixture was concentrated and the residue was triturated with petroleum ether (50 mL) to afford E (3.2 g, 7.06 mmol, 93.49% yield) as a white solid. LCMS (ESI) m / z: [M+H] + = 454.2. 1 H NMR (400MHz, DMSO-d 6 ) δ = 8.22 (s, 1H), 8.15 (d, J = 5.2 Hz, 1H), 7.98 (d, J = 7.6Hz, 1H), 7.79 (s, 1H), 7.69 (d, J = 7.6 Hz, 1H), 7.57-7.54 (m, 1H), 6.96 (d, J = 5.2 Hz, 1H), 6.91 (s, 1H), 5.64-5.54 (m, 2H), 4.30 (s, 2H), 3.44 (s, 2H),3.14 (s, 3H), 1.38 (s, 9H) ppm。

[1262] Step 6: Preparation of Intermediate 122 - 2-((4-(3-(2-((2-((4,5-Dimethyl-1-(methylsulfonyl)pyrrol-3-carbonyl)amino)acetyl)amino)thiazol-4-yl)phenyl)pyridin-2-yl)methylamino)acetic acid (I-95) Step 7: Preparation of N-(2-((4-(3-(2-((2-(4-(1-(2-(2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxyacetyl)-4-piperidinyl)-1-piperidinyl)-2-oxoethyl)methylamino)-4-pyridinyl)phenyl)thiazol-2-yl)amino)-2-oxoethyl)-4,5-dimethyl-1-(methylsulfonyl)pyrrole-3-carboxamide

[1263]

[1264] A mixture of 4,5-dimethyl-1-methylsulfonyl-pyrrole-3-carboxylic acid (E, 263.44 mg, 1.21 mmol), HATU (628.75 mg, 1.65 mmol) and DIEA (427.43 mg, 3.31 mmol, 576.05 µL) in DMF (10 mL) was stirred at 30 °C for 0.5 h. Then I-84 (500 mg, 1.10 mmol) was added and the mixture was stirred for an additional 2 h. The reaction mixture was poured into water (100 mL), filtered and purified by column chromatography (SiO 2 , petroleum ether / EtOAc 5:1 to 1:1) to afford F (430 mg, yield 54.91%, purity 91.9%) as a white solid. LCMS (ESI) m / z:[M+H] + = 653.2. 1 H NMR (400 MHz, DMSO-d 6) δ = 12.38 (s, 1H), 8.48-8.47 (m, 1H), 8.22 (s, 1H), 8.16 (d, J J = 5.2 Hz, 1H), 7.98 (d, J J = 7.6 Hz, 1H), 7.80 (s, 2H), 7.70 (d, J J = 8.0 Hz, 1H), 7.59-7.57 (m, 1H), 6.96-6.94 (m, 1H), 6.91 (s, 1H), 4.30 (s, 2H), 4.11 (d, J J = 5.6 Hz, 2H), 3.48 (s, 3H), 3.14 (s, 3H), 2.32 (s, 3H), 2.12 (s, 3H), 1.39 (s, 9H) ppm。

[1265] (Compound 123) Step 1: Preparation of tert-Butyl 2-((4-(3-(2-((2-((1-(Isopropylsulfonyl)-4,5-dimethylpyrrol-3-carbonyl)amino)acetyl)amino)thiazol-4-yl)phenyl)pyridin-2-yl)methylamino)acetate (B)

[1266]

[1267] A mixture of F (430 mg, 0.66 mmol) in HCl (6 M, 20 mL) was stirred at 30 °C for 15 h. The reaction mixture was filtered, and the filter cake was triturated with petroleum ether / EtOAc (10:1) to give I-95 (260 mg, 57.35% yield) as a grey solid. LCMS (ESI) m / z: [M+H] + = 597.2. 1 1H NMR (400 MHz, DMSO-d 6 ) δ = 12.44 (br s, 1H), 8.54-8.51 (m, 1H), 8.34 (s, 1H), 8.16 (d, J J = 6.4 Hz, 1H), 8.11 (d, J J = 7.6 Hz, 1H), 7.88-7.87 (m, 2H), 7.81 (s, 1H), 7.67-7.63 (m, 1H), 7.51 (s, 1H), 7.39 (d, J J = 6.4 Hz, 1H), 4.70 (s, 2H), 4.11 (d, J J = 5.6 Hz, 2H), 3.49 (s, 3H), 2.32 (s, 3H), 2.12 (s, 3H) ppm。

[1268] Step 2: Preparation of 2-((4-(3-(2-((2-((1-(Isopropylsulfonyl)-4,5-dimethylpyrrol-3-carbonyl)amino)acetyl)amino)thiazol-4-yl)phenyl)pyridin-2-yl)methylamino)acetic acid (C) Step 3: Preparation of N-(2-((4-(3-(2-((2-(5-((2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)pentylamino)-2-oxoethyl)methylamino)-4-pyridinyl)phenyl)thiazol-2-yl)amino)-2-oxoethyl)-1-(isopropylsulfonyl)-4,5-dimethylpyrrole-3-carboxamide (Compound 91) Step 1: Preparation of 1-(1-Isocyanatoethylsulfonyl)-4-methylbenzene (B) Step 2: Preparation of tert-Butyl 5-methyl-1H-pyrrole-3-carboxylate (C)

[1269]

[1270] To a mixture of I-95 (50 mg, 0.079 mmol), I-87 (49.99 mg, 0.083 mmol) and DIEA (43.32 mg, 0.335 mmol, 58.38 μL) in DMF (1 mL) was added EDCI (20.88 mg, 0.11 mmol) and HOBt (14.72 mg, 0.11 mmol), and the mixture was stirred at 30 °C for 2 h. The reaction mixture was added to water (8 mL), and the precipitate was collected by filtration. The solid was purified by preparative HPLC (column: Waters Xbridge 150 x 25 mm x 5 μm; mobile phase: [10 mM NH 4 HCO 3 aqueous solution / ACN]; B%: 27% - 60%, 11 min) to give compound 123 as an orange solid (10.42 mg, 8.91 μmol, 11.28% yield). LCMS (ESI) m / z: [M+H] + = 1061.5. 1 H NMR (400 MHz, DMSO-d 6 ) δ = 12.51 - 12.30 (m, 1H), 11.12 (s, 1H), 8.50 (m, 1H), 8.21 (s, 1H), 8.12 (d, J J = 5.3 Hz, 1H), 7.97 (d, J J = 7.6 Hz, 1H), 7.82 - 7.74 (m, 3H), 7.67 (d, J J = 7.6 Hz, 1H), 7.58 - 7.52 (m, 1H), 7.45 (d, J J = 7.1 Hz, 1H), 7.31 (d, J J = 8.6 Hz, 1H), 6.89 (d, J= 5.3 Hz, 1H), 6.82(s, 1H), 5.28 - 5.01 (m, 3H), 4.61 - 4.44 (m, 2H), 4.40 - 4.28 (m, 2H), 4.13 - 4.05(m, 2H), 3.99 - 3.89 (m, 1H), 3.86 - 3.76 (m, 1H), 3.48 (s, 3H), 3.09 (s, 3H), 3.03 - 2.93 (m, 2H), 2.92 - 2.83 (m, 1H), 2.64 - 2.54 (m, 3H), 2.46 - 2.44 (m, 1H), 2.31 (s, 3H), 2.11 (s, 3H), 2.08 - 1.97 (m, 1H), 1.79 - 1.61 (m, 4H), 1.40 - 1.12(m, 4H), 1.08 - 0.89 (m, 2H) ppm。

[1271]

[1272] Compound 91 - N - [2 - [[4 - [3 - [2 - [[2 - [5 - [[2 - (2,6 - dioxo - 3 - piperidinyl)-1,3 - dioxoisoindolin - 4 - yl]amino]pentylamino]-2 - oxo - ethyl]-methyl - amino]-4 - pyridinyl]phenyl]thiazol - 2 - yl]amino]-2 - oxo - ethyl]-1 - isopropylsulfonyl - 4,5 - dimethyl - pyrrole - 3 - carboxamide

[1273]

[1274] Step 3: Preparation of tert-Butyl 5-methyl-1-(methylsulfonyl)pyrrole-3-carboxylate (D) Step 4: Preparation of 5-Methyl-1-(methylsulfonyl)pyrrole-3-carboxylic acid (E)

[1275]

[1276] To a solution of 1-isopropylsulfonyl-4,5-dimethylpyrrole-3-carboxylic acid (A, 416.44 mg, 1.70 mmol) in DMF (15 mL) was added HATU (880.25 mg, 2.32 mmol) and DIPEA (997.34 mg, 7.72 mmol, 1.34 mL). The mixture was stirred at 30 °C for 0.5 h, then 2-[[4-[3-[2-[(2-aminoacetyl)amino]thiazol-4-yl]phenyl]-2-pyridinyl]-methylamino]tert-butyl acetate (prepared according to the method of I-84, 700 mg, 1.54 mmol) was added. The reaction mixture was stirred at 30 °C for 2.5 h, then poured into water (150 mL) and filtered. The filter cake was purified by column chromatography (SiO 2 , petroleum ether / EtOAc 5:1 to 1:1) to give B (630 mg, 54.36% yield) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 681.2. 1 H NMR (400 MHz, DMSO-d 6 ) δ = 12.41(s, 1H), 8.54-8.21 (m, 1H), 8.22-8.21 (m, 1H), 8.15 (d, J = 5.2 Hz, 1H),7.98-7.96 (m, 1H), 7.81 (s, 1H), 7.77 (s, 1H), 7.70-7.68 (m, 1H), 7.58-7.54(m, 1H), 6.95-6.94 (m, 1H), 6.90 (s, 1H), 4.29 (s, 2H), 4.09 (d, J = 5.6 Hz,2H), 3.76-3.73 (m, 1H), 3.13 (s, 3H), 2.29 (s, 3H), 2.11 (s, 3H), 1.38 (s,9H), 1.23 (s, 3H), 1.22 (s, 3H) ppm。

[1277] Step 5: Preparation of 2-(Methyl-(4-(3-(2-((2-((5-Methyl-1-(methylsulfonyl)pyrrol-3-carbonyl)amino)acetyl)amino)thiazol-4-yl)phenyl)pyridin-2-yl)methylamino)acetic acid ​

[1278]

[1279] A mixture of B (620 mg, 0.91 mmol) in HCl (6 M, 20 mL) was stirred at 30 °C for 15 h. The mixture was filtered and then triturated with petroleum ether / EtOAc (10:1) to give C (320 mg, 47.83% yield) as a pink solid. LCMS (ESI) m / z: [M+H] + = 625.2. 1 H NMR (400 MHz, DMSO-d 6 ) δ = 12.44 (br s, 1H), 8.55 (d, J = 5.6 Hz, 1H), 8.33 (s, 1H), 8.15 (d, J = 6.8 Hz, 1H), 8.09 (d, J = 7.6 Hz, 1H), 7.89 - 7.86 (m, 2H), 7.77 (s, 1H), 7.65 (d, J = 7.6 Hz, 1H), 7.49 (s, 1H), 7.38 (d, J = 6.4 Hz, 1H), 4.68 (s, 2H), 4.10 (sd, J = 5.2 Hz, 2H), 3.77 - 3.71 (m, 1H), 3.32 (s, 3H), 2.29 (s, 3H), 2.11 (s, 3H), 1.23 (s, 3H), 1.22 (s, 3H) ppm.

[1280] ​ ​ ​

[1281]

[1282] To a solution of C (25 mg, 0.040 mmol), 4-(5-aminopentylamino)-2-(2,6-dioxo-3-piperidinyl)isoindoline-1,3-dione (I-10, 18.90 mg, 0.041 mmol), and DIEA (25.86 mg, 0.220 mmol, 34.85 µL) in DMF (0.5 mL) was added HOBt (6.49 mg, 0.048 mmol) and EDCI (9.21 mg, 0.048 mmol), and the mixture was stirred at 30 °C for 2 h. The reaction mixture was concentrated, and the residue was purified by reverse-phase flash chromatography (water / ACN) to give compound 91 (16.77 mg, 41.44% yield) as a yellow solid. LCMS (ESI) m / z: + = 965.7. 1 H NMR (400 MHz, DMSO-d 6 ) δ = 12.81 - 12.02 (m, 1H), 11.08 - 11.07 (m, 1H), 8.52 - 8.49 (m, 1H), 8.22 - 8.20 (m, 1H), 8.14 (d, J = 5.2 Hz, 1H), 7.96 (d, J = 8.0 Hz, 1H), 7.86 - 7.81 (m, 1H), 7.78 - 7.76 (m, 2H), 7.67 (d, J = 7.6 Hz, 1H), 7.59 - 7.52 (m, 2H), 7.06 - 6.99 (m, 2H), 6.92 - 6.89 (m, 1H), 6.84 (s, 1H), 6.48 - 6.45 (m, 1H), 5.10 - 4.98 (m, 1H), 4.18 (s, 2H), 4.09 (d, J = 6.0 Hz, 2H), 3.77 - 3.71 (m, 1H), 3.23 - 3.18 (m, 2H), 3.14 - 3.12 (m, 3H), 3.10 - 3.05 (m, 2H), 2.59 - 2.54 (m, 2H), 2.29 (s, 3H), 2.11 (s, 3H), 2.04 - 1.98 (m, 1H), 1.56 - 1.49 (m, 2H), 1.46 - 1.40 (m, 3H), 1.33 - 1.26 (m, 2H), 1.23 (d, J = 6.8 Hz, 6H) ppm.

[1283]

[1284]

[1285] Preparation of 2-[methyl-[4-[3-[2-[[2-[(5-methyl-1-methylsulfonyl-pyrrole-3-carbonyl)amino]acetyl]amino]thiazol-4-yl]phenyl]-2-pyridinyl]amino]acetic acid (I-102)

[1286]

[1287] ​

[1288]

[1289] At 0 °C, benzyl(triethyl)ammonium chloride (7.00 g, 30.73 mmol) and MeI (43.62 g, 307.32 mmol, 19.13 mL) were added to a solution of 1-(isocyanatomethylsulfonyl)-4-methylbenzene (A, 30 g, 153.66 mmol) in DCM (300 mL). NaOH (307.29 g, 2.30 mol, 30% in water) was added slowly and the reaction mixture was stirred at 0 °C for 4 h. The reaction mixture was diluted with water (300 mL) and extracted with DCM (300 mL x 3). The combined organic layers were washed with water (300 mL x 3), dried over anhydrous Na 2 SO 4 and B was obtained as a brown oil under reduced pressure (31 g, 81.36% yield, 84.396% purity). 1 1H NMR (400 MHz, chloroform-d) δ = 7.89 (d, J = 8.0 Hz, 2H), 7.45 (d, J = 7.6 Hz, 2H), 4.63 - 4.57 (m, 1H), 2.51 (s, 3H), 1.76 (d, J = 6.8 Hz, 3H) ppm.

[1290] ​

[1291]

[1292] To a solution of tert-butyl acrylate (18.99 g, 148.14 mmol, 21.50 mL) and B (31 g, 148.14 mmol) in THF (400 mL) at 40 °C was slowly added NaH (7.11 g, 177.77 mmol, 60% in mineral oil). The reaction mixture was heated to 70 °C and stirred for 1 h. The reaction mixture was poured into saturated aqueous NH 4 Cl (500 mL) and extracted with EtOAc (300 mL x 3). The combined organic phases were washed with brine (300 mL), dried over anhydrous Na 2 SO 4 and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / EtOAc 1:0 - 10:1) to give C as a yellow oil (5.1 g, 20.09 mmol, 13.56% yield, 71.40% purity). LCMS (ESI) m / z: [M - 56 + H] + = 126.1. 1 H NMR (400 MHz, chloroform - d) δ = 8.08 (s, 1H), 7.23 - 7.21 (m, 1H), 6.26 (s, 1H), 2.25 (s, 3H), 1.54 (s, 9H) ppm.

[1293] ​

[1294]

[1295] To a mixture of C (4.6 g, 25.38 mmol) in THF (80 mL) at 0 °C was slowly added NaHMDS (1 M, 50.76 mL). The reaction mixture was stirred for 30 min and then methanesulfonyl chloride (4.36 g, 38.07 mmol, 2.95 mL) was added dropwise at 0 °C. The reaction mixture was stirred at 15 °C for an additional 16 h and then slowly poured into saturated aqueous NH 4 Cl (150 mL) and extracted with EtOAc (100 mL x 3). The combined organic layers were washed with brine (60 mL), dried over anhydrous Na 2 SO 4 and concentrated under reduced pressure. The residue was purified by reverse - phase column chromatography (0.1% aqueous ammonia) to give D as a white solid (3.1 g, 47.10%). 11H NMR (400 MHz, chloroform-d) δ = 7.63 (d, J = 2.0 Hz, 1H), 6.40 - 6.39 (m, 1H), 3.19 (s, 3H), 2.44 (d, J = 0.8 Hz, 3H), 1.54 (s, 9H) ppm.

[1296] ​

[1297]

[1298] A mixture of D (3.1 g, 11.95 mmol) in HCl / 1,4-dioxane (4 M, 50 mL) was stirred at 15 °C for 48 h. The reaction mixture was concentrated under reduced pressure, and the residue was triturated with MTBE (15 mL) and petroleum ether (15 mL) at 15 °C for 10 minutes. The mixture was filtered to give E (2.35 g, 96.74% yield) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 204.1. 1 1H NMR (400 MHz, methanol-d4) δ = 7.69 (d, J J = 1.6 Hz, 1H), 6.41 (s, 1H), 3.35 (s, 3H), 2.44 (d, J J = 0.8 Hz, 3H) ppm.

[1299] ​ Preparation of tert-butyl [[4-[(2-pyridinylamino)phenyl]-1,3-thiazol-2-yl]amino]acetate (F)

[1300]

[1301] At 30 °C, to a mixture of E (224.02 mg, 1.10 mmol) in DCM (5 mL) was added DIEA (427.43 mg, 3.31 mmol, 576.05 µL), HATU (628.75 mg, 1.65 mmol) and tert-butyl 2-[[4-[3-[2-[(2-aminoacetyl)amino]thiazol-4-yl]phenyl]-2-pyridinyl]methyl-amino]acetate (500 mg, 1.10 mmol). The reaction mixture was stirred for 16 h and then washed with water (10 mL x 3). The organic layer was concentrated under reduced pressure and the residue was triturated with EtOAc (10 mL) to give F (550 mg, 76.58% yield) as a yellow solid. LCMS (ESI) m / z: [M+H]+ = 639.3. 1 H NMR (400 MHz, DMSO-d 6 ) δ = 12.40 (s, 1H), 8.62 - 8.58 (m, 1H), 8.26 (s, 1H), 8.15 (d, J = 6.0 Hz, 1H), 8.03 (d, J = 7.6 Hz, 1H), 7.81 (s, 1H), 7.77 (d, J = 7.2 Hz, 1H), 7.74 (d, J = 2.0 Hz, 1H), 7.63 - 7.59 (m, 1H), 7.16 (s, 2H), 6.48 (s, 1H), 4.45 (s, 2H), 4.12 (d, J = 6.0 Hz, 2H), 3.53 (s, 3H), 3.21 (s, 3H), 2.41 (s, 3H), 1.40 (s, 9H) ppm。

[1302] Step 6: 2-[Methyl-[4-[3-[2-[[2-[(5-methyl-1-methylsulfonyl-1H-pyrrole-3-carbonyl)amino]acetyl yl]amino]-1,3-thiazol-4-yl]phenyl]-2-pyridinyl]amino]acetic acid (I-102)

[1303]

[1304] A mixture of F (510 mg, 798.43 µm) in HCl (12 M, 5 mL) was stirred at 25 °C for 20 minutes. The reaction mixture was filtered to give I - 102 as a yellow solid (340 mg, 68.78%). LCMS (ESI) m / z: [M + H] + = 583.0. 1 H NMR (400 MHz, DMSO-d 6 ) δ = 12.42 (s, 1H), 8.63 - 8.60 (m, 1H), 8.31 (s, 1H), 8.14 (d, J = 6.4 Hz, 1H), 8.08 (d, J = 7.6 Hz, 1H), 7.89 - 7.82 (m, 2H), 7.74 (d, J = 2.0 Hz, 1H), 7.67 - 7.62 (m, 1H), 7.44 (s, 1H), 7.33 (d, J = 5.6 Hz, 1H), 6.48 (s, 1H), 4.61 (s, 2H), 4.12 (d, J = 5.6 Hz, 2H), 3.53 (s, 3H), 3.29 (s, 3H), 2.41 (s, 3H) ppm。

[1305] Preparation of Compound 122 - N-(2 - ((4-(3-(2 - ((2 - ((2-(2-(2 - ((2-(2,6 - dioxopiperidin - 3 - yl)-1,3 - dioxoisoindolin - 4 - yl)amino)ethoxy)ethoxy)ethyl)amino)-2 - oxoethyl)(methyl)amino)pyridin - 4 - yl)phenyl)thiazol - 2 - yl)amino)-2 - oxoethyl)-5 - methyl - 1-(methylsulfonyl)-1H - pyrrole - 3 - carboxamide

[1306]

[1307] To a mixture of 2 - [methyl - [4 - [3 - [2 - [[2 - [(5 - methyl - 1 - methylsulfonyl - pyrrole - 3 - carbonyl)amino]acetyl]amino]thiazol - 4 - yl)phenyl]-2 - pyridinyl]amino]acetic acid (I - 102, 40 mg, 0.064 mmol) and 4 - [2 - [2 - (2 - aminoethoxy)ethoxy]ethylamino]-2 - (2,6 - dioxo - 3 - piperidinyl)isoindoline - 1,3 - dione (I - 9, 26.13 mg, 0.064 mmol) in DMF (1 mL) was added DIEA (41.75 mg, 0.32 mmol, 56.27 µL). The mixture was stirred at 30 °C for 15 minutes, then EDCI (18.58 mg, 0.096 mmol) and HOBt (13.10 mg, 0.096 mmol) were added. After stirring for another 16 hours, the mixture was poured into water (10 mL) and extracted with EtOAc (10 mL×3). The combined organic extracts were washed with brine (20 mL), dried over Na 2 SO 4 and concentrated. The residue was purified by preparative HPLC (column: Waters Xbridge 150 x 25 mm x 5 µm; mobile phase: [water (10 mM NH 4 HCO 3 ) - ACN]; B%: 26% - 56%, 10 min) to give Compound 122 (9.9 mg, 14.48%) as a yellow solid. LCMS (ESI) m / z: [M + H] + = 969.5. 1 H NMR (400 MHz, DMSO - d 6 ) δ = 12.44 - 12.36 (m, 1 H), 11.11 (d, J= 4.4 Hz, 1 H), 8.63 - 8.60 (m, 1 H), 8.19 (s, 1 H), 8.13 (d, J = 5.2 Hz, 1 H), 7.96 (d, J = 8.0 Hz, 1 H), 7.90 - 7.87 (m, 1 H), 7.78 (s, 1 H), 7.74 (d, J = 2.0 Hz, 1 H), 7.67 (d, J = 7.6 Hz, 1 H), 7.58 - 7.52 (m, 2H), 7.11 (d, J = 8.8 Hz, 1 H), 7.03 (d, J = 7.2 Hz, 1 H), 6.92 - 6.90 (m, 1 H), 6.85 (s, 1 H), 6.60 - 6.57 (m, 1 H), 6.48 (s, 1 H), 5.06 - 5.02 (m, 1 H), 4.20 (s, 2 H), 4.11 (d, J = 5.6 Hz, 2 H), 3.58 - 3.56 (m, 2 H), 3.53 (s, 3 H), 3.50 - 3.48 (m, 4 H), 3.45 - 3.38 (m, 4 H), 3.23 - 3.19 (m, 2 H), 3.16 (d, J = 5.2 Hz, 1H), 3.11 (s, 3 H), 2.90 - 2.81 (m, 1 H), 2.58 (d, J = 2.4 Hz, 1 H), 2.40 (s, 3H), 2.07 - 1.96 (m, 1 H) ppm。

[1308]

[1309]

[1310] Preparation of N - [2 - [[4 - [3 - [2 - (2 - aminoethoxy) - 4 - pyridyl]phenyl]thiazol - 2 - yl]amino] - 2 - oxo - ethyl] - 1 - mesyl - pyrrole - 3 - carboxamide (I - 96)

[1311]

[1312] Step 1: Preparation of tert-butyl N-[2-[(4-bromo-2-pyridinyl)oxy]ethyl]carbamate (B)

[1313]

[1314] At 0 °C, PPh 3 (1.13 g, 4.31 mmol) and N-(2-hydroxyethyl)carbamic acid tert-butyl ester (555.87 mg, 3.45 mmol, 534.49 µL) were added to a solution of 4-bromopyridin-2-ol (A, 0.5 g, 2.87 mmol) in THF (5 mL). DIAD (871.62 mg, 4.31 mmol, 838.10 µL) was added dropwise and the mixture was stirred at 20 °C for 2 h. The reaction mixture was quenched with water (10 mL) and extracted with DCM (20 mL). The combined organic layers were washed with water (10 mL), dried over Na 2 SO 4 and concentrated under reduced pressure. The residue was purified by column chromatography (SiO 2 , petroleum ether / EtOAc 3:1) to give B (0.6 g, 1.89 mmol, 65.83%) as a white solid. LCMS (ESI) m / z: 81 BrM+H] + = 318.9. 1 H NMR (400 MHz, chloroform-d) δ = 1.46 (s, 9H), 3.54 - 3.52 (m, 2H), 4.38 - 4.36 (m, 2H), 4.95 (s, 1H), 6.96 (d, J = 1.6 Hz, 1H), 7.06 - 7.04 (m, 1H), 7.97 (d, J = 5.6 Hz, 1H) ppm.

[1315] Step 2: Preparation of tert-butyl N-[2-[[4-[3-[2-[[2-[(1-methylsulfonyl-1H-pyrrole-3-carbonyl)amino]acetyl]amino]-1,3-thia zol-4-yl]phenyl]-2-pyridinyl]oxy]ethyl]carbamate (C)

[1316]

[1317] 1-Methanesulfonyl-N-[2-oxo-2-[[4-[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]thiazol-2-yl]amino]ethyl]pyrrole-3-carboxamide (I-90, 530 mg, 1.0 mmol), B (380.31 mg, 1.20 mmol), K 3 PO 4 (636.30 mg, 3.00 mmol) and Pd(dppf)Cl 2A mixture of (73.11 mg, 0.1 mmol) in 1,4-dioxane (6 mL) / water (2 mL) was degassed and purged with nitrogen three times. The mixture was stirred at 80 °C for 2 h and then concentrated under reduced pressure. The residue was purified by column chromatography (SiO 2 , petroleum ether / EtOAc 1:1) to give C (0.4 g, 62.48%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 641.2.

[1318] 1 H NMR (400 MHz, DMSO-d 6 ) δ = 1.39 (s, 9H), 3.58 (s, 3H), 4.16 - 4.14(m, 2H), 4.33 - 4.31 (m, 2H), 6.79 - 6.77 (m, 1H), 7.01 (s, 1H), 7.16 (s, 1H),7.34 - 7.30 (m, 1H), 7.38 - 7.36 (m, 1H), 7.59 - 7.57 (m, 1H),7.75 - 7.73 (m, 1H),7.85 (s, 2H), 8.01 (d, J = 7.6 Hz, 1H), 8.25 (d, J = 5.2 Hz, 1H), 8.30 (s, 1H),8.70 - 8.68 (m, 1H),12.44 (s, 1H) ppm。

[1319] Step 3: Preparation of 1-methylsulfonyl-N-[2-oxo-2-[[4-[3-(2-piperazin-1-yl-4-pyridinyl)phenyl]-1,3-thiazol-2- yl]amino]ethyl]-1H-pyrrole-3-carboxamide (I-96)

[1320]

[1321] To a solution of C (300 mg, 0.4618 mmol) in DCM (0.5 mL) was added HCl / 1,4-dioxane (4 M, 2.34 mL). The mixture was stirred at 20 °C for 2 h and then concentrated under reduced pressure. The residue was suspended in MTBE (10 mL), filtered, and triturated with TBME (3 mL) to give crude I-96 (0.25 g, 92.53% yield) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 541.1. 1 H NMR (400 MHz, DMSO-d 6) δ = 12.44 (s, 1H), 8.71 (s, 1H), 8.33 - 8.26 (m, 2H), 8.03 (d, J = 7.8 Hz, 3H), 7.85 (d, J = 6.0 Hz, 2H), 7.73 (d, J = 8.4 Hz, 1H), 7.64 - 7.57 (m, 1H), 7.45 (d, J = 5.2 Hz, 1H), 7.35 - 7.30 (m, 1H), 7.19 (s, 1H), 6.78 (s, 1H), 4.57 - 4.47 (m, 2H), 4.16 - 4.14 (m, 2H), 3.58 (s, 3H), 3.29 - 3.25 (m, 2H) ppm。

[1322] Preparation of Compound 36 - N - [2 - [[4 - [3 - [2 - [2 - [4 - [2 - (2,6 - dioxo - 3 - piperidinyl) - 1,3 - dioxoisoindolin - 4 - yl]oxybutanoyl amino]ethoxy] - 4 - pyridyl]phenyl]thiazol - 2 - yl]amino] - 2 - oxo - ethyl] - 1 - mesyl - pyrrole - 3 - carboxamide

[1323]

[1324] To a solution of N - [2 - [[4 - [3 - [2 - (2 - aminoethoxy) - 4 - pyridyl]phenyl]thiazol - 2 - yl]amino] - 2 - oxo - ethyl] - 1 - mesyl - pyrrole - 3 - carboxamide (I - 96, 30 mg, 0.052 mmol), 4 - [2 - (2,6 - dioxo - 3 - piperidinyl) - 1,3 - dioxoisoindolin - 4 - yl]oxybutyric acid (I - 11, 22.48 mg, 0.06 mmol), EDCI (19.93 mg, 0.103 mmol) and HOBt (14.05 mg, 0.103 mmol) in DMF (1 mL) was added DIEA (33.59 mg, 0.259 mmol, 45.28 μL). The mixture was stirred at 25 °C for 16 h. The reaction mixture was filtered and the filtrate was purified directly by preparative HPLC (column: Phenomenex Synergi C18 150 x 25 x 10μm; mobile phase: [0.225% aqueous formic acid / ACN: 38% - 68%, 9 min), to give Compound 36 (14.03 mg, 28.94% yield) as a white solid.

[1325] LCMS (ESI) m / z: [M + H] += 883.2. 1 H NMR (400 MHz, DMSO-d 6 ) δ = 8.76 - 8.74 (m, 1H), 8.32 - 8.22 (m, 2H), 8.16 - 8.14 (m, 1H), 8.00 (d, J J = 7.6 Hz, 1H), 7.89 - 7.69 (m, 4H), 7.60 - 7.30 (m, 5H), 7.14 (s, 1H), 6.79 - 6.78 (m, 1H), 5.09 - 5.06 (m, 1H), 4.40 - 4.13 (m, 6H), 3.59 (s, 3H), 3.51 - 3.47 (m, 2H), 2.94 - 2.81 (m, 1H), 2.62 - 2.53 (m, 2H), 2.34 - 2.32 (m, 2H), 2.06 - 1.94 (m, 3H) ppm。

[1326]

[1327]

[1328]

[1329]

[1330]

[1331] Preparation of 1 - (Methylsulfonyl) - N - [2 - oxo - 2 - [[4 - [3 - (2 - piperazin - 1 - yl - 4 - pyridinyl)phenyl]thiazol - 2 - yl]amino]ethyl]pyrrole - 3 - carboxamide (I - 97)

[1332]

[1333] Step 1: Preparation of tert-butyl 4-(4-bromo-2-pyridinyl)piperazine-1-carboxylate (B)

[1334]

[1335] To a solution of 4-bromo-2-fluoropyridine (A, 2 g, 11.36 mmol) and tert-butyl piperazine-1-carboxylate (4.23 g, 22.73 mmol) in DMSO (20 mL) was added DIPEA (7.34 g, 56.82 mmol, 9.90 mL). The mixture was stirred at 120 °C for 16 h, then poured into water (50 mL) and extracted with EtOAc (50 mL x 3). The combined organic extracts were washed with brine (100 mL), dried over Na 2 SO 4 and concentrated to give B (3.5 g, 85.86%) as a brown solid, which was used without further purification. LCMS (ESI) m / z: 81 BrM+H] + = 343.9. 1 H NMR (400 MHz, DMSO-d 6 ) δ = 7.99 (d, J = 5.2 Hz, 1H), 7.06 (s, 1H), 6.85 - 6.83 (m, 1H), 3.54 - 3.48 (m, 4H), 3.41 - 3.39 (m, 4H), 1.42 (s, 9H) ppm.

[1336] Step 2: Preparation of tert-butyl 4-[4-[3-[2-[[2-[(1-methylsulfonyl-1H-pyrrole-3-carbonyl)amino]acetyl]amino]-1,3-thia zol-4-yl]phenyl]-2-pyridinyl]piperazine-1-carboxylate (C)

[1337]

[1338] Under a nitrogen atmosphere, to a solution of 1-(methylsulfonyl)-N-[2-oxo-2-[[4-[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]thiazol-2-yl]amino]ethyl]pyrrole-3-carboxamide (I-90, 800 mg, 1.51 mmol) and B (774.24 mg, 2.26 mmol) in 1,4-dioxane (8 mL) / water (0.8 mL) was added Pd(dppf)Cl 2 (110.36 mg, 0.150 mmol) and K 3 PO 4 (960.44 mg, 4.52 mmol). The mixture was stirred at 100 °C for 4 h, then diluted with water (15 mL) and extracted with EtOAc (20 mL x 3). The combined organic extracts were washed with brine (20 mL x 2), dried over anhydrous Na 2 SO 4Dry and concentrate under reduced pressure. Purify the residue by column chromatography (SiO 2 , petroleum ether / EtOAc 5:1 to 1:2) to obtain C as a yellow solid (650 mg, 57.96%, 89.533% purity). LCMS (ESI) m / z: [M+H] + = 666.1.

[1339] Step 3: Preparation of 1-methylsulfonyl-N-[2-oxo-2-[[4-[3-(2-piperazin-1-yl-4-pyridinyl)phenyl]-1,3-thiazol-2- yl]amino]ethyl]-1H-pyrrole-3-carboxamide (I-97)

[1340]

[1341] Stir a solution of C (400 mg, 0.660 mmol) in 4 M HCl / 1,4-dioxane (4 mL) at 20 °C for 1 h. Concentrate the reaction mixture under reduced pressure and stir the residue with MTBE (5 mL) for 5 minutes. Filter the solid and triturate it with MTBE (3 mL x 3) to obtain I-97 as a brown solid (120 mg, 23.92%, 72.112% purity), which can be used without further purification. LCMS (ESI) m / z: [M+H] + = 566.1. 1 H NMR (400 MHz, methanol-d4) δ = 8.36 (s, 1H), 8.15 - 8.09 (m, 2H), 7.86 - 7.82 (m, 2H), 7.71 (s, 1H), 7.65 (s, 1H), 7.63 - 7.59 (m, 1H), 7.51 (d, J = 6.4 Hz, 1H), 7.29 - 7.28 (m, 1H), 6.83 - 6.82 (m, 1H), 4.28 (s, 2H), 4.14 - 4.11 (m, 4H), 3.55 - 3.51 (m, 4H), 3.39 (s, 3H) ppm.

[1342] Preparation of Compound 31 - N-[2 - [[4 - [3 - [2 - [4 - [3 - [2 - [2 - [[2 - (2,6 - dioxo - 3 - piperidinyl)-1,3 - dioxoisoindolin - 4 - yl]amino]ethoxy]ethoxy]propanoyl]piperazin - 1 - yl]-4 - pyridyl]phenyl]thiazol - 2 - yl]amino]-2 - oxo - ethyl]-1 - mesyl - pyrrole - 3 - carboxamide

[1343]

[1344] At 10 °C, to a solution of 3-[2-[2-[[2-(2,6-dioxo-3-piperidinyl)-1,3-dioxoisoindolin-4-yl]amino]ethoxy]ethoxy]propanoic acid (I-17, 17.99 mg, 0.041 mmol) in DMF (0.4 mL) was added EDCI (11.94 mg, 0.062 mmol) and HOBt (8.42 mg, 0.062 mmol). 1-Mesyl-N-[2-oxo-2-[[4-[3-(2-piperazin-1-yl-4-pyridinyl)phenyl]thiazol-2-yl]amino]ethyl]pyrrole-3-carboxamide (I-97, 25 mg, 0.041 mmol) and DIPEA (16.10 mg, 0.124 mmol, 21.70 μL) were added and the mixture was stirred at 25 °C for 2.5 h. The reaction mixture was purified directly by preparative HPLC (column: Waters Xbridge 150 x 25mm x 5μm; mobile phase: 4 HCO 3 aqueous solution / ACN] 26%-56%, 10 min) to give compound 31 (15.33 mg, 37.64%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 981.3. 1 H NMR (400 MHz, DMSO-d 6 ) δ = 12.44 - 12.31 (m, 1H), 11.12 - 11.05 (m, 1H), 8.66 - 8.64 (m, 1H), 8.21 - 8.20 (m, 2H), 7.98 - 7.76 (m, 1H), 7.85 - 7.84 (m, 1H), 7.79 (s, 1H), 7.70 - 7.68 (m, 1H), 7.57 - 7.53 (m, 2H), 7.32 - 7.30 (m, 1H), 7.12 - 7.10 (m, 2H), 7.02 - 6.99 (m, 2H), 6.78 - 6.76 (m, 1H), 6.60 - 6.57 (m, 1H), 5.06 - 5.02 (m, 1H), 4.14 - 4.13 (d, J = 6.0 Hz, 2H), 3.67 - 3.52 (m, 19H), 3.46 - 3.42 (m, 3H), 2.90 - 2.82 (m, 2H), 2.62 - 2.59 (m, 4H), 2.05 - 1.98 (m, 2H) ppm.

[1345]

[1346]

[1347]

[1348]

[1349]

[1350] Preparation of 2-Methyl-2-[3-[3-[2-[[2-[(1-Methanesulfonylpyrrole-3-carbonyl)amino]acetyl]amino]thiazol-4-yl]phenyl]pyrazol-1-yl]propanoic Acid (I-98)

[1351]

[1352] Step 1: Preparation of tert-butyl 2-(3-bromopyrazol-1-yl)-2-methylpropanoate (B)

[1353]

[1354] At -60 °C, LDA (2 M in THF, 2.63 mL) was added to a solution of tert-Butyl 2-(3-bromopyrazol-1-yl)acetate (A, 550 mg, 2.11 mmol) in THF (5 mL). The mixture was stirred for 30 minutes and then a solution of MeI (747.43 mg, 5.27 mmol, 327.82 μL) in THF (0.5 mL) was added. The reaction mixture was warmed to 25 °C and stirred for 1 hour, then quenched with saturated aqueous NH 4 Cl (5 mL) and extracted with EtOAc (5 mL x 3). The combined organic extracts were washed with brine (5 mL x 3) and concentrated under reduced pressure. The residue was purified by flash silica chromatography (ISCO®; 24 g SepaFlash® Silica Flash Column, eluent 0 - 50% EtOAc / petroleum ether gradient, 35 mL / min) to give B (350 mg, 55.16%) as a colorless oil. LCMS (ESI) m / z: 79 Br M-56+H] + =233.1. 1 H NMR (400 MHz, DMSO-d 6 ) δ = 7.91 (d, J = 2.4 Hz, 1H), 6.42 (d, J =2.4 Hz, 1H), 1.71 (s, 6H), 1.34 (s, 9H) ppm。

[1355] Step 2: Preparation of tert-butyl 2-methyl-2-[3-[3-[2-[[2-[(1-methylsulfonyl-1H-pyrrole-3-carbonyl)amino]acetyl]am ino]-1,3-thiazol-4-yl]phenyl]pyrazol-1-yl]propanoate (C)

[1356]

[1357] 1-Mesyl-N-[2-oxo-2-[[4-[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]thiazol-2-yl]amino]ethyl]pyrrole-3-carboxamide (I-90, 580 mg, 1.09 mmol), B (350 mg, 1.16 mmol), di-tert-butyl(cyclopentyl)phosphine palladium(II) dichloride (142.53 mg, 0.21 mmol), and K 3 PO 4 (696.32 mg, 3.28 mmol) in a mixture of 1,4-dioxane (5 mL) / water (1 mL) was degassed and purged with nitrogen three times. The mixture was stirred at 80 °C for 2 hours, then diluted with water (10 mL) and extracted with EtOAc (10 mL × 3). The combined organic layers were washed with brine (10 mL). The organic phase was concentrated under reduced pressure and the residue was triturated with EtOAc (20 mL) and petroleum ether (5 mL) for 10 minutes and filtered. The filter cake was dried under reduced pressure to give C (500 mg, 68.66%) as a white solid. LCMS (ESI) m / z: [M+H] + = 613.5. 1 H NMR (400 MHz, DMSO-d 6 ) δ = 12.43 (brs, 1H), 8.70 - 8.68 (m, 1H), 8.33 (s, 1H), 7.95 (d, J = 2.4 Hz, 1H), 7.87 - 7.80 (m, 2H), 7.75 (d, J = 7.6 Hz, 1H), 7.69 (s, 1H), 7.47 - 7.45 (m, 1H), 7.33 - 7.31 (m, 1H), 6.83 - 6.75 (m, 2H), 4.16 (d, J = 5.6 Hz, 2H), 3.58 (s, 3H), 1.79 (s, 6H), 1.35 (s, 9H) ppm。

[1358] Step 3: Preparation of 2-methyl-2-[3-[3-[2-[[2-[(1-methylsulfonyl-1H-pyrrole-3-carbonyl)amino]acetyl]am ino]-1,3-thiazol-4-yl]phenyl]pyrazol-1-yl]propanoic acid (I-98)

[1359]

[1360] To a solution of C (500 mg, 0.816 mmol) in DCM (10 mL) at 25 °C was added TFA (3.85 g, 33.77 mmol, 2.5 mL). The mixture was stirred at 25 °C for 16 h and then concentrated under reduced pressure. The residue was triturated with EtOAc (20 mL) and filtered to give I-98 (460 mg, 99.05% yield) as a white solid. LCMS (ESI) m / z: [M+H] + = 557.3. 1 H NMR (400 MHz, DMSO-d 6 ) δ = 12.48 (s, 1H), 8.71 - 8.69 (m, 1H), 8.36 (s, 1H), 7.97 (d, J = 2.4 Hz, 1H), 7.88 - 7.79 (m, 2H), 7.75 - 7.73 (m, 1H), 7.70 (s, 1H), 7.47 - 7.45 (m, 1H), 7.33 - 7.31 (m, 1H), 6.79 (d, J = 2.4 Hz, 2H), 4.16 - 4.14 (m, 2H), 3.58 (s, 3H), 1.81 (s, 6H) ppm.

[1361] Preparation of 6-N-(2-((4-(3-(1-(1-((2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethyl)amino)-2-methyl-1-oxopropan-2-yl)-1H-pyrazol-3-yl)phenyl)thiazol-2-yl)amino...

Claims

1. A compound having a structure of Formula I: ALB Formula I, in L is a linker, wherein the linker has the following structure: in x1 is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14; x2 is 0, 1, 2, 3, 4, 5, or 6; x3 is 1 or 2; x4 is 1 or 2; W is R x1 and R x2 are each independently H, halogen, C1-C6 alkyl, C1-C6 heteroalkyl or C3-C6 carbocyclyl, or R x1 and R x2 Together with the carbon atoms to which they are attached, they form a C3-C6 carbocyclyl or a C2-C5 heterocyclyl; and R y1 , R y2 , R y3 and R y4 Each is independently H, C1-C6 alkyl, C1-C6 heteroalkyl or C3-C6 carbocyclyl; B is a degradation moiety, wherein the degradation moiety is a ubiquitin ligase binding moiety comprising the following structure: and A has the following structure: in G 1 It is C6-C 10 Arylene, C2-C9 heterocyclic group or C 2- C9 heteroarylene; G 2 is absent, -O-, C1-C6 alkylene, C1-C6 alkenylene, C1-C6 heteroalkylene, C2-C9 heterocyclylC1-C6 alkylene, or C2-C9 heteroarylC1-C6 alkylene; G 3 Does not exist, C6-C 10 Arylene, C6-C 10 Cycloalkylene, C2-C9 heterocyclylene, or C2-C9 heteroarylene; and A 1 is the bond between A and the linker; or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1, wherein G 2 Does not exist.

3. The compound of claim 1, wherein G 1 yes 4. The compound of claim 1, wherein G 3 Does not exist.

5. The compound of claim 1, wherein G 3 yes 6. The compound of claim 1, wherein G 3 yes 7. The compound according to claim 1, wherein the linker has the following structure:

8. Compounds having the structure of Formula I: ALB Formula I, in L is a linker, wherein the linker has the following structure: B is a degradation moiety, wherein the degradation moiety is a ubiquitin ligase binding moiety comprising the following structure: and A has the following structure: in G 1 It is C6-C 10 Arylene, C2-C9 heterocyclic group or C 2- C9 heteroarylene; G 2 is absent, -O-, C1-C6 alkylene, C1-C6 alkenylene, C1-C6 heteroalkylene, C2-C9 heterocyclylC1-C6 alkylene, or C2-C9 heteroarylC1-C6 alkylene; G 3 Does not exist, C6-C 10 Arylene, C6-C 10 Cycloalkylene, C2-C9 heterocyclylene, or C2-C9 heteroarylene; and A 1 is the bond between A and the linker; or a pharmaceutically acceptable salt thereof.

9. A pharmaceutical composition comprising the compound according to any one of claims 1 to 8 and a pharmaceutically acceptable excipient.

10. Use of a compound according to any one of claims 1 to 8 for the preparation of a medicament for treating a BAF complex-related disorder in a subject in need thereof.

11. Use of a compound as claimed in any one of claims 1 to 8 for the preparation of a medicament for reducing the level and / or activity of BRG1 and / or BRM in cancer in a subject in need thereof.

12. The method of claim 11, 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.

13. A compound or a pharmaceutically acceptable salt thereof, wherein the compound has the following structure:

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