Compounds and uses thereof

Abstract

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

Description

[0001] background

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

[0003] Chromatin regulation is crucial for gene expression, and ATP-dependent chromatin remodeling is a mechanism by which such gene expression occurs. The human switch / sucrose non-fermentable (SWI / SNF) chromatin remodeling complex (also known as the BAF complex) possesses two SWI2-like ATPases, 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 certain cancer tumors and is required for cancer cell proliferation. BRM (also known as the putative global transcriptional activator SNF2L2 and / or the 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 BRG1 loss-of-function mutations. Inactivation of BRG and / or BRM leads to downstream cellular effects, including cell cycle arrest and tumor suppression. Summary of the Invention

[0004] The present invention features compounds that can be used to modulate the BAF complex. In certain embodiments, the compounds can be used to treat disorders associated with alterations in the BAF complex, for example, disorders 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 disorders.

[0005] In one aspect, the invention features a compound having the structure of Formula I or a pharmaceutically acceptable salt thereof:

[0006]

[0007] in

[0008] X 1 Is O or NR 2 ;

[0009] Each X 2 are independently halogen;

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

[0011] m is 0, 1, 2, 3, or 4;

[0012] R 1 is halogen or optionally substituted C1-C6 alkyl;

[0013] R 2 is H or optionally substituted C1-C6 alkyl;

[0014] L 1 is an optionally substituted C1-C6 alkylene group;

[0015] L is included A linker of a structure;

[0016] n is 0, 1, 2, or 3;

[0017] L 2 is an optionally substituted C1-C6 alkylene, an optionally substituted C1-C 20 heteroalkylene or optionally substituted C2-C9 heterocyclylene;

[0018] Each L 3 are independently -O-, optionally substituted C1-C 20 Heteroalkylene, optionally substituted C3-C 10 Carbocyclylene, optionally substituted C3-C 10 Carbocyclylene-C1-C 20 an alkylene group, an optionally substituted C2-C9 heterocyclylene group, or an optionally substituted C2-C9 heterocyclylene group-C1-C 20 an alkylene group; and

[0019] D is the degradation fraction.

[0020] In certain embodiments, m is 0. In certain embodiments, m is 1 or 2.

[0021] In certain embodiments, X 1 Is O. In certain embodiments, X 1 It is NR 2 .

[0022] In certain embodiments, R 2 is an optionally substituted C1-C6 alkyl. 2 In certain embodiments, R 2 It's methyl.

[0023] In certain embodiments, L 1 yes .

[0024] In certain embodiments, L2 is an optionally substituted C1-C6 alkylene or an optionally substituted C1-C 20 In certain embodiments, L 2 is an optionally substituted C2-C9 heterocyclylene group.

[0025] In certain embodiments, L 2 is an optionally substituted C1-C6 alkylene. In certain embodiments, L 2 is optionally substituted C1-C 20 Heteroalkylene.

[0026] In certain embodiments, L 2 yes or In certain embodiments, L 2 yes .

[0027] In certain embodiments, L 2 yes

[0028]

[0029]

[0030] .

[0031] In certain embodiments, n is 1. In certain embodiments, n is 2. In certain embodiments, n is 3.

[0032] In certain embodiments, each L 3 is independently optionally substituted C1-C 20 Heteroalkylene, optionally substituted C3-C 10 Carbocyclylene, optionally substituted C3-C 10 In certain embodiments, each L is a C1-C6 alkylene, an optionally substituted C2-C9 heterocyclylene, or an optionally substituted C2-C9 heterocyclylene-C1-C6 alkylene. 3 is independently optionally substituted C3-C 10 carbocyclylene-C1-C6 alkylene, optionally substituted C2-C9 heterocyclylene, or optionally substituted C2-C9 heterocyclylene-C1-C6 alkylene.

[0033] In certain embodiments, each L 3 Independently

[0034]

[0035]

[0036] .

[0037] In certain embodiments, each L 3 Independently

[0038] .

[0039] In certain embodiments, n is 0.

[0040] In certain embodiments, k is 0, 1, or 2. In certain embodiments, each X 2 is independently fluoro or chloro.

[0041] In certain embodiments, the compound or a pharmaceutically acceptable salt thereof has Formula Ib:

[0042] .

[0043] In one aspect, the invention features a compound having the structure of Formula II or a pharmaceutically acceptable salt thereof:

[0044]

[0045] in

[0046] A Z 1 and a Z 2 to form an optionally substituted C1-C4 alkylene group, and the remaining Z 1 and Z 2 Each is hydrogen;

[0047] Each X 2 are independently halogen;

[0048] k is 0, 1, 2, 3, or 4;

[0049] L is A linker of a structure;

[0050] q is 0, 1, 2, 3, or 4;

[0051] L 4 is an optionally substituted C1-C6 alkylene, an optionally substituted C1-C 20 heteroalkylene or optionally substituted C2-C9 heteroarylene;

[0052] Each L 5 are independently -O-, optionally substituted C1-C6 alkylene, optionally substituted C1-C 20Heteroalkylene, optionally substituted C3-C 10 Carbocyclylene, optionally substituted C3-C 10 Carbocyclylene-C1-C6 alkylene, optionally substituted C2-C9 heterocyclylene or C2-C9 heterocyclylene-C1-C 20 an alkylene group; and

[0053] D is the degradation fraction.

[0054] In certain embodiments, the compound or a pharmaceutically acceptable salt thereof has Formula IIa:

[0055] .

[0056] In certain embodiments, the compound or a pharmaceutically acceptable salt thereof has Formula IIb:

[0057]

[0058] in

[0059] L 4 is an optionally substituted C1-C6 alkylene or an optionally substituted C1-C 20 heteroalkylene;

[0060] L 5 Absent, optionally substituted C3-C 10 carbocyclylene-C1-C6 alkylene or optionally substituted C2-C9 heterocyclylene-C1-C6 alkylene; and

[0061] D is the degradation fraction.

[0062] In certain embodiments, q is 1. In certain embodiments, q is 2. In certain embodiments, q is 3. In certain embodiments, q is 4.

[0063] In certain embodiments, L is .

[0064] In certain embodiments, L 4 is an optionally substituted C1-C6 alkylene. In certain embodiments, L 4 is optionally substituted C1-C 20 Heteroalkylene.

[0065] In certain embodiments, L 4 yes or In certain embodiments, L 4 yes

[0066]

[0067] .

[0068] In certain embodiments, L 5 In certain embodiments, each L 5 are independently -O-, optionally substituted C1-C6 alkylene, optionally substituted C1-C 20 Heteroalkylene, optionally substituted C3-C 10 Carbocyclylene, optionally substituted C3-C 10 In certain embodiments, each L is a C1-C6 alkylene or an optionally substituted C2-C9 heterocyclylene-C1-C6 alkylene. 5 is independently optionally substituted C3-C 10 carbocyclylene-C1-C6 alkylene or optionally substituted C2-C9 heterocyclylene-C1-C6 alkylene.

[0069] In certain embodiments, (L 5 ) q yes

[0070] In certain embodiments, (L 5 ) q yes .

[0071] In certain embodiments, the compound or a pharmaceutically acceptable salt thereof has Formula IIb:

[0072] .

[0073] In certain embodiments of any of the aforementioned compounds, the degradation moiety is a ubiquitin ligase binding moiety.

[0074] In certain embodiments, the ubiquitin ligase binding moiety comprises a Cereblon ligand, an IAP (inhibitor of apoptosis) ligand, mouse double minute 2 homolog (MDM2), or a von Hippel-Lindau ligand, or a derivative or analog thereof.

[0075] In certain embodiments, the degradation moiety comprises a structure of Formula Y or a pharmaceutically acceptable salt thereof:

[0076]

[0077] in

[0078] A 2 is the bond between the degradable moiety and the linker;

[0079] v1 is 0, 1, 2, 3, 4, or 5;

[0080] u1 is 1, 2, or 3;

[0081] T 1 Is a key or ;

[0082] T 2 yes ;

[0083] R 5A is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl;

[0084] Each R J1 is independently halogen, optionally substituted C1-C6 alkyl or optionally substituted C1-C6 heteroalkyl;

[0085] J A is absent, is O, optionally substituted amino, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; and

[0086] J is absent and is optionally substituted C3-C 10 Carbocyclylene, optionally substituted C6-C 10 an arylene group, an optionally substituted C2-C9 heterocyclylene group, or an optionally substituted C2-C9 heteroarylene group.

[0087] In certain embodiments, T 2 yes In certain embodiments, T 2 yes or In certain embodiments, T 2 yes In certain embodiments, T 2 yes .

[0088] In certain embodiments, the degradation moiety comprises a structure of Formula Y1:

[0089]

[0090] or a pharmaceutically acceptable salt thereof.

[0091] In certain embodiments, T 1 In certain embodiments, T 1 yes .

[0092] In certain embodiments, the degradation moiety comprises a structure of Formula Y2:

[0093]

[0094] or a pharmaceutically acceptable salt thereof.

[0095] In certain embodiments, the degradation moiety comprises a structure of Formula Z:

[0096]

[0097] or a pharmaceutically acceptable salt thereof.

[0098] In certain embodiments, u1 is 1. In certain embodiments, u1 is 2. In certain embodiments, u1 is 3.

[0099] In certain embodiments, the degradation moiety comprises a structure of Formula AA0:

[0100]

[0101] or a pharmaceutically acceptable salt thereof.

[0102] In certain embodiments, the degradation moiety comprises a structure of Formula AB:

[0103]

[0104] or a pharmaceutically acceptable salt thereof.

[0105] In certain embodiments, the degradation moiety comprises the structure of Formula AC:

[0106]

[0107] or a pharmaceutically acceptable salt thereof.

[0108] In certain embodiments, J A In certain embodiments, J A is an optionally substituted C1-C6 alkyl. In certain embodiments, J A is an optionally substituted C1-C6 heteroalkyl. A is O or optionally substituted amino.

[0109] In certain embodiments, J A yes .

[0110] In certain embodiments, the degradation moiety comprises a structure of Formula AA0:

[0111]

[0112] or a pharmaceutically acceptable salt thereof.

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

[0114] In certain embodiments, the degradation moiety comprises a structure of Formula AA1:

[0115]

[0116] or a pharmaceutically acceptable salt thereof.

[0117] In certain embodiments, the degradation moiety comprises a structure of Formula AB1:

[0118]

[0119] or a pharmaceutically acceptable salt thereof.

[0120] In certain embodiments, the degradation moiety comprises a structure of Formula AC1:

[0121]

[0122] or a pharmaceutically acceptable salt thereof.

[0123] In certain embodiments, J is absent. In certain embodiments, J is optionally substituted C3-C 10 carbocyclylene or optionally substituted C6-C 10 In certain embodiments, J is an optionally substituted C2-C9 heterocyclylene or an optionally substituted C2-C9 heteroarylene.

[0124] In certain embodiments, J is an optionally substituted heterocyclylene. In certain embodiments, J is an optionally substituted C6-C 10 Arylene.

[0125] In certain embodiments, J is .

[0126] In certain embodiments, the degradation moiety comprises a structure of Formula AA2:

[0127]

[0128] or a pharmaceutically acceptable salt thereof.

[0129] In certain embodiments, the degradation moiety comprises a structure of Formula AA3:

[0130]

[0131] or a pharmaceutically acceptable salt thereof.

[0132] In certain embodiments, the degradation moiety comprises a structure of Formula AA4:

[0133]

[0134] or a pharmaceutically acceptable salt thereof.

[0135] In certain embodiments, R A5 is H or optionally substituted C1-C6 alkyl. A5 is optionally substituted C1-C6 heteroalkyl.

[0136] In certain embodiments, R A5 Is H or methyl. In certain embodiments, R A5 is H. In certain embodiments, R A5 In certain embodiments, R A5 yes .

[0137] In certain embodiments, the degradation moiety comprises a structure of Formula A or a pharmaceutically acceptable salt thereof:

[0138]

[0139] in

[0140] Y 1 yes ;

[0141] R A5 is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl;

[0142] R A6 is H or optionally substituted C1-C6 alkyl; and R A7 is H or optionally substituted C1-C6 alkyl; or R A6 and R A7 Together with the carbon atoms to which they are attached, they form an optionally substituted C3-C6 carbocyclyl or an optionally substituted C2-C5 heterocyclyl; or R A6 and R A7 Combined together with the carbon atoms to which they are attached to form an optionally substituted C3-C6 carbocyclyl or an optionally substituted C2-C5 heterocyclyl;

[0143] R A8 is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl;

[0144] R A1 、R A2 、R A3 and R A4 Each of which is independently H, A 2 , halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, optionally substituted -O-C3-C6 carbocyclyl, hydroxyl, thiol, or optionally substituted amino; or R A1 and R A2 、R A2 and R A3 , and / or R A3 and R A4 Combined with the carbon atoms to which they are attached to form ;and is optionally substituted C6-C 10 Aryl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heteroaryl or C2-C9 heterocyclyl, any of which is optionally substituted by A 2 Substituted, where R A1 、R A2 、R A3 and R A4 One of them is A 2 ,or A 2 replace.

[0145] In certain embodiments, R A1 、R A2 、R A3 and R A4 Each of which is independently H, A 2 , halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxy, thiol, or optionally substituted amino; or R A1 and R A2 、R A2 and R A3, and / or R A3 and R A4 Combined with the carbon atoms to which they are attached to form ;and is optionally substituted C6-C 10 Aryl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heteroaryl or C2-C9 heterocyclyl, any of which is optionally substituted by A 2 Substituted, where R A1 、R A2 、R A3 and R A4 One of them is A 2 ,or A 2 substituted, or a pharmaceutically acceptable salt thereof.

[0146] In certain embodiments, R A1 、R A2 、R A3 and R A4 Each of H, A 2 , halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted -O-C3-C6 carbocyclyl, hydroxy, optionally substituted amino; or R A1 and R A2 、R A2 and R A3 , or R A3 and R A4 Combined with the carbon atoms to which they are attached to form ;and is an optionally substituted C2-C9 heterocyclic group, which is optionally substituted by A 2 Substituted, where R A1 、R A2 、R A3 and R A4 One of them is A 2 ,or A 2 replace.

[0147] In certain embodiments, R A1 、R A2 、R A3 and R A4 Each of which is independently H, A 2 ,F, ; or R A1 and R A2 、R A2 and R A3 , or R A3 and RA4 Combined with the carbon atoms to which they are attached to form ;and is an optionally substituted C2-C9 heterocyclic group, which is optionally substituted by A 2 Substituted, where R A1 、R A2 、R A3 and R A4 One of them is A 2 ,or A 2 replace.

[0148] In certain embodiments, R A1 It's A 2 In certain embodiments, R A2 It's A 2 In certain embodiments, R A3 It's A 2 In certain embodiments, R A4 It's A 2 In certain embodiments, R A5 It's A 2 .

[0149] In certain embodiments, R A1 、R A2 、R A3 and R A4 Each of is independently H or A 2 .

[0150] In certain embodiments, R A1 It's A 2 And R A2 、R A3 and R A4 Each of is H. In certain embodiments, R A2 It's A 2 And R A1 、R A3 and R A4 Each of is H. In certain embodiments, R A3 It's A 2 And R A1 、R A2 and R A4 Each of is H. In certain embodiments, R A4 It's A 2 And R A1 、R A2 and R A3 Each of is H.

[0151] In certain embodiments, R A5is H or optionally substituted C1-C6 alkyl. A5 Is H or In certain embodiments, R A5 is H. In certain embodiments, R A5 yes .

[0152] In certain embodiments, R A8 is H or optionally substituted C1-C6 alkyl. A8 Is H or In certain embodiments, R A8 is H. In certain embodiments, R A8 yes .

[0153] In certain embodiments, Y 1 yes .

[0154] In certain embodiments, Y 1 yes In certain embodiments, Y 1 yes or .

[0155] In certain embodiments, Y 1 yes In certain embodiments, Y 1 yes In certain embodiments, Y 1 yes In certain embodiments, Y 1 yes .

[0156] In certain embodiments, R A6 and R A7 Each of which is independently H, F, ; or R A6 and R A7 Combined with the carbon atoms to which they are attached to form In certain embodiments, R A6 is H and R A7 It’s H.

[0157] In certain embodiments, Y 1 yes In certain embodiments, Y 1 yes In certain embodiments, Y 1 yes .

[0158] In certain embodiments, the degradation moiety comprises a structure of Formula A1:

[0159]

[0160] or a pharmaceutically acceptable salt thereof.

[0161] In certain embodiments, the degradation moiety comprises the structure of Formula A2:

[0162]

[0163] or a pharmaceutically acceptable salt thereof.

[0164] In certain embodiments, the degradation moiety comprises a structure of Formula A3:

[0165]

[0166] or a pharmaceutically acceptable salt thereof.

[0167] In certain embodiments, the degradation moiety comprises the structure of Formula A4:

[0168]

[0169] or a pharmaceutically acceptable salt thereof.

[0170] In certain embodiments, the degradation moiety comprises the structure of Formula A5:

[0171]

[0172] or a pharmaceutically acceptable salt thereof.

[0173] In certain embodiments, the degradation moiety comprises the structure of Formula A6:

[0174]

[0175] or a pharmaceutically acceptable salt thereof.

[0176] In certain embodiments, the degradation moiety comprises the structure of Formula A7:

[0177]

[0178] or a pharmaceutically acceptable salt thereof.

[0179] In certain embodiments, the degradation moiety comprises the structure of Formula A8:

[0180]

[0181] or a pharmaceutically acceptable salt thereof.

[0182] In certain embodiments, the degradation moiety comprises the structure of Formula A9:

[0183]

[0184] or a pharmaceutically acceptable salt thereof.

[0185] In certain embodiments, the degradation moiety comprises the structure of Formula A10:

[0186]

[0187] or a pharmaceutically acceptable salt thereof.

[0188] In certain embodiments, wherein the degradation moiety comprises

[0189]

[0190]

[0191] or its derivatives or analogs.

[0192] In certain embodiments, the degradation moiety comprises structure.

[0193] In certain embodiments, the degradation moiety comprises or its derivatives or analogs.

[0194] In certain embodiments, wherein the degradation moiety comprises In certain embodiments, the degradation portion comprises In certain embodiments, the degradation portion comprises structure.

[0195] In certain embodiments, yes or , where R A9 It is H, A 2 , optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl.

[0196] In certain embodiments, the degradation moiety comprises

[0197]

[0198] structure.

[0199] In certain embodiments, R A9 It is H, A 2 or optionally substituted C1-C6 alkyl. In certain embodiments, R A9 It is H, A 2 In certain embodiments, R 9A is H. In certain embodiments, R 9A In certain embodiments, R A9 It's A 2 .

[0200] In certain embodiments, the degradation moiety comprises structure.

[0201] In certain embodiments, the degradation moiety comprises a structure of Formula B or a pharmaceutically acceptable salt thereof:

[0202]

[0203] in

[0204] R A5 is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl;

[0205] R A1 、R A2 、R A3 and R A4 Each of which is independently H, A 2 , halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, optionally substituted -O-C3-C6 carbocyclyl, hydroxyl, thiol, or optionally substituted amino; or R A1 and R A2 、R A2 and R A3 , and / or R A3 and R A4 Combined with the carbon atoms to which they are attached to form ;and is optionally substituted C6-C 10 Aryl, optionally substituted C3-C10 carbocyclyl, optionally substituted C2-C9 heteroaryl or C2-C9 heterocyclyl, any of which is optionally substituted by A 2 Substituted, where R A1 、R A2 、R A3 and R A4 One of them is A 2 ,or A 2 replace.

[0206] In certain embodiments, R A1 、R A2 、R A3 and R A4 Each of H, A 2 , halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted -O-C3-C6 carbocyclyl, hydroxy, optionally substituted amino; or R A1 and R A2 、R A2 and R A3 , or R A3 and R A4 Combined with the carbon atoms to which they are attached to form ;and is an optionally substituted C2-C9 heterocyclic group, which is optionally substituted by A 2 Substituted, where R A1 、R A2 、R A3 and R A4 One of them is A 2 ,or A 2 replace.

[0207] In certain embodiments, R A1 、R A2 、R A3 and R A4 Each of which is independently H, A 2 ,F, ; or R A1 and R A2 、R A2 and R A3 , or R A3 and R A4 Combined with the carbon atoms to which they are attached to form ;and is an optionally substituted C2-C9 heterocyclic group, which is optionally substituted by A 2 Substituted, where R A1 、R A2 、RA3 and R A4 One of them is A 2 ,or A 2 replace.

[0208] In certain embodiments, R A1 It's A 2 In certain embodiments, R A2 It's A 2 In certain embodiments, R A3 It's A 2 In certain embodiments, R A4 It's A 2 In certain embodiments, R A5 It's A 2 .

[0209] In certain embodiments, R A5 is H or optionally substituted C1-C6 alkyl.

[0210] In certain embodiments, R A5 Is H or In certain embodiments, R A5 is H. In certain embodiments, R A5 yes .

[0211] In certain embodiments, the degradation moiety comprises a structure of Formula Bl:

[0212]

[0213] or a pharmaceutically acceptable salt thereof.

[0214] In certain embodiments, the degradation moiety comprises the structure of Formula B2:

[0215]

[0216] or a pharmaceutically acceptable salt thereof.

[0217] In certain embodiments, the degradation moiety comprises a structure of Formula B3:

[0218]

[0219] or a pharmaceutically acceptable salt thereof.

[0220] In certain embodiments, the degradation moiety comprises the structure of Formula B4:

[0221]

[0222] or a pharmaceutically acceptable salt thereof.

[0223] In certain embodiments, the degradation moiety comprises In certain embodiments, the degradation portion comprises In certain embodiments, the degradation portion comprises structure.

[0224] In certain embodiments, the ubiquitin ligase binding moiety comprises a von Hippel-Lindau ligand.

[0225] In certain embodiments, the von Hippel-Lindau ligand comprises The structure of A 2 is a bond between the degradable portion and the linker, or a derivative or analog thereof.

[0226] In certain embodiments, the degradation moiety comprises a structure of Formula C:

[0227]

[0228] in

[0229] L 6 Yes-N(R B1 )(R B2 ), ;

[0230] R B1 It is H, A 2 , optionally substituted C1-C6 alkyl or optionally substituted C1-C6 heteroalkyl;

[0231] R B2 is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl;

[0232] R B3 It's A 2 , optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 Carbocyclyl, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 aryl;

[0233] R B4 is H, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10Carbocyclyl, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 aryl;

[0234] R B5 is H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl;

[0235] v2 is 0, 1, 2, 3, or 4;

[0236] Each R B6 Independently A 2 , halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxy, thiol, or optionally substituted amino;

[0237] R B7 and R B8 Each of the alkyl groups is independently H, halogen, optionally substituted C1-C6 alkyl or optionally substituted C6-C 10 aryl;

[0238] R B9 is H or optionally substituted C1-C6 alkyl; and

[0239] A 2 is the bond between the degradable moiety and the linker;

[0240] where R B1 、R B3 and R B6 One and only one of them is A 2 .

[0241] In certain embodiments, the degradation moiety has the structure of Formula C1:

[0242] .

[0243] In certain embodiments, the degradation moiety has the structure of Formula C2:

[0244] .

[0245] In certain embodiments, R B9is an optionally substituted C1-C6 alkyl. B9 In certain embodiments, R B9 Bonded to ( S )-stereocenter. In certain embodiments, R B9 It's hydrogen.

[0246] In certain embodiments, the degradation moiety has the following structure: .

[0247] In certain embodiments, the degradation moiety has the following structure:

[0248] .

[0249] In certain embodiments, the degradation moiety has the following structure:

[0250] .

[0251] In certain embodiments, the degradation moiety has the following structure:

[0252] .

[0253] In certain embodiments, the degradation moiety is

[0254] .

[0255] In certain embodiments, the degradation portion comprises a structure

[0256] In certain embodiments, the degrader moiety comprises a structure of Formula D or a pharmaceutically acceptable salt thereof:

[0257]

[0258] in

[0259] A 2 is the bond between B and the linker;

[0260] R C1 、R C2 and R C7 Each of wherein is independently H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl;

[0261] R C3 is an optionally substituted C1-C6 alkyl, an optionally substituted C3-C 10 Carbocyclyl, optionally substituted C6-C 10Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 aryl;

[0262] R C5 is an optionally substituted C1-C6 alkyl, an optionally substituted C3-C 10 Carbocyclyl, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 aryl;

[0263] v3 is 0, 1, 2, 3, or 4;

[0264] Each R C8 are independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxy, thiol, or optionally substituted amino;

[0265] v4 is 0, 1, 2, 3, or 4; and

[0266] Each R C9 are independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxyl, thiol or optionally substituted amino.

[0267] In certain embodiments, the degradation moiety comprises a structure of the formula:

[0268] .

[0269] In certain embodiments, the degradation moiety comprises a structure of Formula E or a pharmaceutically acceptable salt thereof:

[0270]

[0271] in

[0272] A 2 is the bond between B and the linker;

[0273] R C10 and R C11 Each of the alkyl groups is independently H, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10 Carbocyclyl, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 aryl;

[0274] v5 is 0, 1, 2, 3, or 4;

[0275] Each R C12 are independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxy, thiol, or optionally substituted amino;

[0276] v6 is 0, 1, 2, 3, or 4; and

[0277] Each R 21 are independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxyl, thiol or optionally substituted amino.

[0278] In certain embodiments, the degradation moiety comprises a structure of the formula:

[0279] .

[0280] In certain embodiments, the degradation moiety comprises a structure of Formula FA or a pharmaceutically acceptable salt thereof:

[0281]

[0282] in

[0283] yes , or by A 2 substituted by one or more independently selected from H, R FF1 and a bicyclic moiety substituted with an oxo group;

[0284] Is a single bond or a double bond;

[0285] u2 is 0, 1, 2, or 3;

[0286] A 2 is the bond between the degradation agent and the linker;

[0287] Y Fa It's CR Fb R Fc , C═O, C═S, C═CH2, SO2, S(O), P(O)Oalkyl, P(O)NHalkyl, P(O)N(alkyl)2, P(O)alkyl, P(O)OH, P(O)NH2;

[0288] Y Fb NH, NR FF1 , CH2, CHR FF1 、C(R FF1 )2, O or S;

[0289] Y Fc It's CR Fd R Fe , C═O, C═S, C═CH2, SO2, S(O), P(O)Oalkyl, P(O)NHalkyl, P(O)N(alkyl)2, P(O)alkyl, P(O)OH, P(O)NH2;

[0290] R Fb 、R Fc 、R Fd and R Fe where each is independently H, alkyl, aliphatic, heteroaliphatic, aryl, heteroaryl, carbocyclyl, hydroxy, alkoxy, amino, -NHalkyl, or -Nalkyl;

[0291] or R Fb and R Fc Together with the carbon atoms to which they are attached, they form a 3-, 4-, 5-, or 6-membered spirocarbocyclylene, or a 4-, 5-, or 6-membered spiroheterocyclylene comprising 1 or 2 heteroatoms selected from N and O;

[0292] or R Fd and R FeTogether with the carbon atoms to which they are attached, they form a 3-, 4-, 5-, or 6-membered spirocarbocyclylene, or a 4-, 5-, or 6-membered spiroheterocyclylene comprising 1 or 2 heteroatoms selected from N and O; and

[0293] or R Fd and R Fb Together with the carbon atoms to which they are attached, they form a 1-, 2-, 3-, or 4-carbon bridged ring;

[0294] Y Fd and Y Ff Each of which is independently CH2, CHR FF2 、C(R FF2 )2、C(O), N、NH、NR FF3 , O, S or S(O);

[0295] Y Fe Is a key or connected to Y Fd and Y Ff a divalent moiety containing 1 to 5 contiguous carbon atoms forming a 3- to 8-membered ring,

[0296] One, two or three of the carbon atoms may be replaced by nitrogen, oxygen or sulfur atoms;

[0297] wherein one of the ring atoms is replaced by A 2 and the other ring atoms are substituted by one or more independently selected from H and R FF1 and

[0298] where Y Fe Adjacent atoms can be connected by single or double bonds;

[0299] Each R FF1 is independently H, alkyl, alkenyl, alkynyl, aliphatic, heteroaliphatic, carbocyclyl, halogen, hydroxy, amino, cyano, alkoxy, aryl, heteroaryl, heterocyclyl, alkylamino, alkylhydroxy, or haloalkyl;

[0300] Each R FF2 are independently alkyl, alkene, alkyne, halogen, hydroxy, alkoxy, azide, amino,

[0301] -C(O)H, -C(O)OH, -C(O)(aliphatic group, including alkyl group), -C(O)O(aliphatic group, including alkyl group),

[0302] -NH(aliphatic group, including alkyl group), -N(aliphatic group, including alkyl group)(aliphatic group, including alkyl group), -NHSO2alkyl,

[0303] -N(alkyl)SO2alkyl, -NHSO2aryl, -N(alkyl)SO2aryl, -NHSO2alkenyl, -N(alkyl)SO2alkenyl,

[0304] -NHSO2 alkynyl, -N(alkyl)SO2 alkynyl, aliphatic, heteroaliphatic, aryl, heteroaryl, heterocyclic, carbocyclic, cyano, nitro, nitroso, -SH, -S alkyl, or haloalkyl; and

[0305] R FF3 is alkyl, alkenyl, alkynyl, -C(O)H, -C(O)OH, -C(O)alkyl or -C(O)Oalkyl,

[0306] If Y Fd or Y Ff A 2 Substitution, then Y Fe Yes key.

[0307] In certain embodiments, the degradation moiety comprises a structure of Formula FA1:

[0308]

[0309] or a pharmaceutically acceptable salt thereof.

[0310] In certain embodiments, the degradation moiety comprises a structure of Formula FB or a pharmaceutically acceptable salt thereof:

[0311]

[0312] in

[0313] yes , or by A 2 substituted by one or more independently selected from H, R FF1 and a bicyclic moiety substituted with an oxo group;

[0314] A 2 is the bond between the degradation agent and the linker;

[0315] Y Fa It's CR Fb R Fc , C═O, C═S, C═CH2, SO2, S(O), P(O)Oalkyl, P(O)NHalkyl, P(O)N(alkyl)2, P(O)alkyl, P(O)OH, P(O)NH2;

[0316] Y Fb and Y Fg Each of NH, NR FF1 , CH2, CHR FF1、C(R FF1 )2, O or S;

[0317] Y Fc It's CR Fd R Fe , C═O, C═S, C═CH2, SO2, S(O), P(O)Oalkyl, P(O)NHalkyl, P(O)N(alkyl)2, P(O)alkyl, P(O)OH, P(O)NH2;

[0318] R Fb 、R Fc 、R Fd 、R Fe 、R Ff and R Fg where each is independently H, alkyl, aliphatic, heteroaliphatic, aryl, heteroaryl, carbocyclyl, hydroxy, alkoxy, amino, -NHalkyl, or -Nalkyl;

[0319] or R Fb and R Fc Together with the carbon atoms to which they are attached, they form a 3-, 4-, 5-, or 6-membered spirocarbocyclylene, or a 4-, 5-, or 6-membered spiroheterocyclylene comprising 1 or 2 heteroatoms selected from N and O;

[0320] or R Fd and R Fe Together with the carbon atoms to which they are attached, they form a 3-, 4-, 5-, or 6-membered spirocarbocyclylene, or a 4-, 5-, or 6-membered spiroheterocyclylene comprising 1 or 2 heteroatoms selected from N and O;

[0321] or R Ff and R Fg Together with the carbon atoms to which they are attached, they form a 3-, 4-, 5-, or 6-membered spirocarbocyclylene, or a 4-, 5-, or 6-membered spiroheterocyclylene comprising 1 or 2 heteroatoms selected from N and O;

[0322] or R Fd and R Fb Together with the carbon atoms to which they are attached, they form a 1-, 2-, 3-, or 4-carbon bridged ring;

[0323] or R Fd and R Ff Together with the carbon atoms to which they are attached, they form a 1-, 2-, 3-, or 4-carbon bridged ring;

[0324] or R Fb and R Fg Together with the carbon atoms to which they are attached, they form a 1-, 2-, 3-, or 4-carbon bridged ring;

[0325] Y Fdand Y Ff Each of which is independently CH2, CHR FF2 、C(R FF2 )2、C(O), N、NH、NR FF3 , O, S or S(O);

[0326] Y Fe Is a key or connected to Y Fd and Y Ff a divalent moiety containing 1 to 5 contiguous carbon atoms forming a 3- to 8-membered ring,

[0327] One, two or three of the carbon atoms may be replaced by nitrogen, oxygen or sulfur atoms;

[0328] wherein one of the ring atoms is replaced by A 2 and the other ring atoms are substituted by one or more independently selected from H and R FF1 and

[0329] where Y Fe Adjacent atoms can be connected by single or double bonds;

[0330] Each R FF1 is independently H, alkyl, alkenyl, alkynyl, aliphatic, heteroaliphatic, carbocyclyl, halogen, hydroxy, amino, cyano, alkoxy, aryl, heteroaryl, heterocyclyl, alkylamino, alkylhydroxy, or haloalkyl;

[0331] Each R FF2 are independently alkyl, alkene, alkyne, halogen, hydroxy, alkoxy, azide, amino,

[0332] -C(O)H, -C(O)OH, -C(O)(aliphatic group, including alkyl group), -C(O)O(aliphatic group, including alkyl group),

[0333] -NH(aliphatic, including alkyl), -N(aliphatic, including alkyl)(aliphatic, including alkyl), -NHSO2alkyl, -N(alkyl)SO2alkyl, -NHSO2aryl, -N(alkyl)SO2aryl, -NHSO2alkenyl, -N(alkyl)SO2alkenyl, -NHSO2alkynyl, -N(alkyl)SO2alkynyl, aliphatic, heteroaliphatic, aryl, heteroaryl, heterocyclic, carbocyclic, cyano, nitro, nitroso, -SH, -Salkyl, or haloalkyl; and

[0334] R FF3 is alkyl, alkenyl, alkynyl, -C(O)H, -C(O)OH, -C(O)alkyl or -C(O)Oalkyl,

[0335] If Y Fdor Y Ff A 2 Substitution, then Y Fe Yes key.

[0336] In certain embodiments, the degradation moiety comprises a structure of Formula FB1:

[0337]

[0338] or a pharmaceutically acceptable salt thereof.

[0339] In certain embodiments, the degradation moiety comprises a structure of Formula F1 or a pharmaceutically acceptable salt thereof:

[0340]

[0341] Among them A 2 is the bond between the degradation agent and the linker; and R F1 Does not exist or is O.

[0342] In certain embodiments, R F1 In certain embodiments, R F1 It's O.

[0343] In certain embodiments, the degradation moiety comprises or structure.

[0344] In certain embodiments, the degradation moiety comprises a structure of Formula F2 or a pharmaceutically acceptable salt thereof:

[0345]

[0346] Among them A 2 is the bond between the degradation agent and the linker; and Y 2 It is CH2 or NH.

[0347] In certain embodiments, Y 2 In certain embodiments, Y 2 It's CH2.

[0348] In certain embodiments, the degradation moiety comprises or structure.

[0349] In certain embodiments, the degradation moiety comprises a structure of Formula G or a pharmaceutically acceptable salt thereof:

[0350]

[0351] Among them A 2is the bond between the degradation agent and the linker; and Y 3 It is CH2 or NH.

[0352] In certain embodiments, Y 3 In certain embodiments, Y 3 It's CH2.

[0353] In certain embodiments, the degradation moiety comprises or structure.

[0354] The degradation moieties may also include structures found in, for example, WO2017 / 197036, WO2019 / 204354, WO2019 / 236483, WO2020 / 010177, and WO2020 / 010227, the structures of which are incorporated herein by reference.

[0355] In certain embodiments, the degradation moiety comprises The structure of A 2 It is the bond between the degradable portion and the linker or a derivative or analog thereof.

[0356] In certain embodiments, the compound has the structure of any one of compounds 1-75 in Table 1, or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound has the structure of any one of compounds 76-104 in Table 2, or a pharmaceutically acceptable salt thereof.

[0357] In one aspect, the invention features a compound having the structure of any one of Compounds 1-75 in Table 1, or a pharmaceutically acceptable salt thereof.

[0358] In one aspect, the invention features a compound having the structure of any one of compounds 105-272 in Table 2, or a pharmaceutically acceptable salt thereof.

[0359] In another aspect, the invention features a compound having the structure of any one of Compounds 76-104 in Table 2, or a pharmaceutically acceptable salt thereof.

[0360] Table 1. Compounds of the present invention

[0361]

[0362]

[0363]

[0364]

[0365]

[0366]

[0367]

[0368]

[0369]

[0370]

[0371]

[0372]

[0373]

[0374]

[0375]

[0376]

[0377]

[0378]

[0379] Table 2. Compounds of the present invention

[0380]

[0381]

[0382]

[0383]

[0384]

[0385]

[0386]

[0387]

[0388]

[0389]

[0390]

[0391]

[0392]

[0393]

[0394]

[0395]

[0396]

[0397]

[0398]

[0399]

[0400]

[0401]

[0402]

[0403]

[0404]

[0405]

[0406]

[0407]

[0408]

[0409]

[0410]

[0411]

[0412]

[0413]

[0414]

[0415]

[0416]

[0417]

[0418]

[0419]

[0420]

[0421]

[0422]

[0423]

[0424]

[0425]

[0426]

[0427]

[0428]

[0429]

[0430]

[0431]

[0432]

[0433]

[0434]

[0435]

[0436]

[0437]

[0438]

[0439]

[0440]

[0441]

[0442]

[0443]

[0444]

[0445]

[0446]

[0447] In one aspect, the invention features a pharmaceutical composition comprising any one of the aforementioned compounds and a pharmaceutically acceptable excipient.

[0448] In another aspect, the invention features a method of reducing the activity of a BAF complex in a cell, comprising contacting the cell with an effective amount of any one of the aforementioned compounds or a pharmaceutical composition thereof.

[0449] In certain embodiments, the cell is a cancer cell.

[0450] In another aspect, the invention features a method of treating a BAF complex-related disorder in a subject in need thereof, comprising administering to the subject an effective amount of any one of the aforementioned compounds (e.g., a BRM / BRG1 dual inhibitor compound or a BRM-selective compound) or a pharmaceutical composition thereof.

[0451] In certain embodiments, the BAF complex-associated disorder is cancer.

[0452] In another aspect, the invention features a method of inhibiting BRM, comprising contacting a cell with an effective amount of any of the aforementioned compounds (eg, a BRM / BRG1 dual inhibitor compound or a BRM-selective compound) or a pharmaceutical composition thereof.

[0453] In certain embodiments, the cell is a cancer cell.

[0454] In another aspect, the invention features a method of inhibiting BRG1, comprising contacting the cell with an effective amount of any one of the aforementioned compounds or a pharmaceutical composition thereof.

[0455] In certain embodiments, the cell is a cancer cell.

[0456] In another aspect, the invention features a method of inhibiting BRM and BRG1, comprising contacting the cell with an effective amount of any one of the aforementioned compounds or a pharmaceutical composition thereof.

[0457] In certain embodiments, the cell is a cancer cell.

[0458] In another aspect, the invention features a method of treating a disorder associated with a BRG1 loss-of-function mutation in a subject in need thereof, comprising administering to the subject an effective amount of any one of the aforementioned compounds (e.g., a BRM / BRG1 dual inhibitor compound or a BRM-selective compound) or a pharmaceutical composition thereof.

[0459] In certain embodiments, the disorder associated with a BRG1 loss-of-function mutation is cancer. In other embodiments, the subject is determined to have a BRG1 loss-of-function disorder, e.g., is 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).

[0460] In another aspect, the invention features a method of inducing apoptosis in a cell, comprising contacting the cell with an effective amount of any of the aforementioned compounds (e.g., a BRM / BRG1 dual inhibitor compound or a BRM-selective compound) or a pharmaceutical composition thereof.

[0461] In certain embodiments, the cell is a cancer cell.

[0462] In another aspect, the invention features a method of treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of any one of the aforementioned compounds (e.g., a BRM / BRG1 dual inhibitor compound or a BRM-selective compound) or a pharmaceutical composition thereof.

[0463] In certain embodiments of any of the foregoing methods, the cancer is non-small cell lung cancer, colorectal cancer, bladder cancer, cancer of unknown primary, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, esophageal-gastric cancer, pancreatic cancer, hepatobiliary cancer, soft tissue sarcoma, ovarian cancer, head and neck cancer, renal cell carcinoma, bone cancer, non-Hodgkin lymphoma, small cell lung cancer, prostate cancer, embryonal carcinoma, germ cell tumor, cervical cancer, thyroid cancer, salivary gland cancer, gastrointestinal neuroendocrine tumor, uterine sarcoma, gastrointestinal stromal tumor, central nervous system cancer, thymic tumor, adrenocortical carcinoma, appendix cancer, small intestine cancer, or penile cancer.

[0464] In certain embodiments of any of the preceding methods, the cancer is non-small cell lung cancer, colorectal cancer, bladder cancer, cancer of unknown primary, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, or penile cancer.

[0465] In certain 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, a CTLA4 inhibitor, a PD1 inhibitor, an interferon therapy, a BRAF inhibitor, a MEK inhibitor, radiation therapy, temozolomide, irinotecan, CAR-T therapy, Herceptin®, Perjeta®, tamoxifen, Xeloda®, docetaxel, a platinum agent such as carboplatin, a taxane such as paclitaxel and docetaxel, an ALK inhibitor, a MET inhibitor, Alimta®, Abraxane®, Doxorubicin®, gemcitabine, Avastin®, Halaven®, neratinib, a PARP inhibitor, ARN810, an mTOR inhibitor, topotecan, Gemzar®, a VEGFR2 inhibitor, a folate receptor antagonist, densizumab, fortabulin, or a PDL1 inhibitor).

[0466] In certain embodiments of any one of the foregoing methods, the cancer has or has been determined to have a BRG1 mutation. In certain embodiments of any one of the foregoing methods, the BRG1 mutation is homozygous. In certain embodiments of any one of the foregoing methods, the cancer does not have or has been determined to have an epidermal growth factor receptor (EGFR) mutation. In certain embodiments of any one of the foregoing methods, the cancer does not have or has been determined to have an anaplastic lymphoma kinase (ALK) driver mutation. In certain embodiments of any one of the foregoing methods, the cancer has or has been determined to have a KRAS mutation. In certain embodiments of any one of the foregoing methods, the BRG1 mutation is in the ATPase catalytic domain of the protein. In certain embodiments of any one of the foregoing methods, the BRG1 mutation is a deletion at the C-terminus of BRG1.

[0467] In another aspect, the present disclosure provides a method for treating a BAF-related disorder (e.g., cancer or viral infection) in a subject in need thereof, comprising contacting a cell with an effective amount of any one of the aforementioned compounds (e.g., a BRM / BRG1 dual inhibitor compound or a BRM-selective compound) or a pharmaceutically acceptable salt thereof, or any one of the aforementioned pharmaceutical compositions. In certain embodiments, the disorder is a viral infection, which is a retroviridae such as a lentivirus (e.g., human immunodeficiency virus (HIV) and a delta retrovirus (e.g., human T-cell leukemia virus I (HTLV-I), human T-cell leukemia virus II (HTLV-II)), a hepadnaviridae (e.g., hepatitis B virus (HBV)), a flaviviridae (e.g., hepatitis C virus (HCV)), adenoviridae (e.g., human adenovirus), a herpesviridae (e.g., human cytomegalovirus (HCMV), Epstein-Barr virus, herpes simplex virus 1 (HSV-1), herpes simplex virus 2 (HSV-2), human herpes virus 6 (HHV-6), herpes virus K*, CMV, varicella-zoster virus), a papillomaviridae (e.g., human papillomavirus (HPV), HPV-1), or a combination thereof. In some embodiments, the disorder is infection with a virus of the family 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 certain embodiments, the disorder is Coffin Siris, neurofibromatosis (e.g., NF-1, NF-2, or schwannomatosis), or multiple meningiomas.

[0468] In another aspect, the present disclosure provides a method for treating a viral infection in a subject in need thereof, comprising administering to the subject an effective amount of any one of the aforementioned compounds (e.g., a BRM / BRG1 dual inhibitor compound or a BRM-selective compound) or a pharmaceutically acceptable salt thereof, or any one of the aforementioned pharmaceutical compositions. In certain embodiments, the viral infection is a retroviridae such as a lentivirus (e.g., human immunodeficiency virus (HIV) and a delta retrovirus (e.g., human T-cell leukemia virus I (HTLV-I), human T-cell leukemia virus II (HTLV-II)), a hepadnaviridae (e.g., hepatitis B virus (HBV)), a flaviviridae (e.g., hepatitis C virus (HCV)), an adenoviridae (e.g., human adenovirus), a herpesviridae (e.g., human cytomegalovirus (HCMV), Epstein-Barr virus, herpes simplex virus 1 (HSV-1), herpes simplex virus 2 (HSV-2), human herpes virus 6 (HHV-6), herpes virus K*, CMV, varicella-zoster virus), a papillomaviridae (e.g., human papillomavirus (HPV), HPV-1), or a combination thereof. E1)), infection with a virus of the family Parvoviridae (e.g., Parvovirus B19), the family Polyomaviridae (e.g., JC virus and BK virus), the family Paramyxoviridae (e.g., Measles virus), or the family Togaviridae (e.g., Rubella virus).

[0469] In certain embodiments of any of the foregoing aspects, the compound is a BRM-selective compound. In certain embodiments, the BRM-selective compound inhibits the level and / or activity of BRM at least 10-fold greater than the level and / or activity of BRG1, 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 certain embodiments, the IC of a BRM-selective compound is 50 or IP 50 It is the IC of BRG1 50 or IP 50 In certain embodiments of any of the foregoing aspects, the compound is a BRM / BRG1 dual inhibitor compound. In certain embodiments, the BRM / BRG1 dual inhibitor compound has similar activity against both BRM and BRG1 (e.g., the compound has an activity against BRM and BRG1 that is within 10-fold (e.g., less than 5-fold, less than 2-fold). In certain embodiments, the activity of the BRM / BRG1 dual inhibitor compound is greater against BRM. In certain embodiments, the activity of the BRM / BRG1 dual inhibitor compound is greater against BRM. For example, in certain embodiments, the IC of the BRM / BRG1 dual inhibitor compound against BRM is greater than that of the BRM / BRG1 dual inhibitor compound. 50 or IP50 The IC of BRG1 50 or IP 50 within 10 times of .

[0470] In another aspect, the invention features a method of treating melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or a hematological cancer in a subject in need thereof, comprising administering to the subject an effective amount of any one of the foregoing compounds or a pharmaceutical composition thereof.

[0471] 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, comprising administering to the subject an effective amount of any one of the foregoing compounds or a pharmaceutical composition thereof.

[0472] In another aspect, the invention features a method of inhibiting metastatic progression of melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or hematological cancer in a subject, comprising administering an effective amount of any one of the foregoing compounds or a pharmaceutical composition thereof.

[0473] In another aspect, the invention features a method of inhibiting metastatic colonization of melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or hematological cancer in a subject, comprising administering an effective amount of any one of the foregoing compounds or a pharmaceutical composition thereof.

[0474] 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, comprising contacting the cells with an effective amount of any one of the foregoing compounds or a pharmaceutical composition thereof.

[0475] In certain embodiments of any of the above aspects, the melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or hematological cell is in a subject.

[0476] In certain embodiments of any of the above aspects, the effective amount of the compound reduces the level and / or activity of BRG1 by at least 5% (e.g., 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) compared to a reference. In certain embodiments, the effective amount of the compound reduces the level and / or activity of BRG1 by at least 50% (e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) compared to a reference. In certain embodiments, the effective amount of the compound reduces the level and / or activity of BRG1 by at least 90% (e.g., 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%).

[0477] In certain embodiments, an effective amount of the compound reduces the level and / or activity of BRG1 by at least 5% (e.g., 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) compared to a reference for at least 12 hours (e.g., 14 hours, 16 hours, 18 hours, 20 hours, 22 hours, 24 hours, 30 hours, 36 hours, 48 ​​hours, 72 hours, or more). In certain embodiments, an effective amount of the compound reduces the level and / or activity of BRG1 by at least 5% (e.g., 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) compared to a reference for at least 4 days (e.g., 5 days, 6 days, 7 days, 14 days, 28 days, or more).

[0478] In certain embodiments of any of the above aspects, an effective amount of the compound reduces the level and / or activity of a BRM by at least 5% (e.g., 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) compared to a reference. In certain embodiments, an effective amount of the compound reduces the level and / or activity of a BRM by at least 50% (e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) compared to a reference. In certain embodiments, an effective amount of the compound reduces the level and / or activity of a BRM by at least 90% (e.g., 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%).

[0479] In certain embodiments, an effective amount of a compound reduces the level and / or activity of a BRM by at least 5% (e.g., 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) compared to a reference for at least 12 hours (e.g., 14 hours, 16 hours, 18 hours, 20 hours, 22 hours, 24 hours, 30 hours, 36 hours, 48 ​​hours, 72 hours, or more). In certain embodiments, an effective amount of a compound reduces the level and / or activity of a BRM by at least 5% (e.g., 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) compared to a reference for at least 4 days (e.g., 5 days, 6 days, 7 days, 14 days, 28 days, or more).

[0480] In certain embodiments, the subject has cancer. In certain embodiments, the cancer expresses BRG1 and / or BRM protein, and / or the cell or subject has been identified as expressing BRG1 and / or BRM. In certain embodiments, the cancer expresses BRG1 protein, and / or the cell or subject has been identified as expressing BRG1. In certain embodiments, the cancer expresses BRM protein, and / or the cell or subject has been identified as expressing BRM. In certain embodiments, the cancer is melanoma (e.g., uveal melanoma, mucosal melanoma, or cutaneous melanoma). In certain embodiments, the cancer is prostate cancer. In certain embodiments, the cancer is a hematological cancer, for example, 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 (for example, T-cell acute lymphoblastic leukemia or B-cell acute lymphoblastic leukemia), diffuse large cell lymphoma or non-Hodgkin lymphoma. In certain embodiments, the cancer is breast cancer (for example, ER-positive breast cancer, ER-negative breast cancer, triple-positive breast cancer or triple-negative breast cancer). In certain embodiments, the cancer is bone cancer (for example, Ewing's sarcoma). In certain embodiments, the cancer is renal cell carcinoma (for example, microphthalmia transcription factor (MITF) family translocation renal cell carcinoma (tRCC)). In certain embodiments, the cancer is metastatic (for example, the cancer has spread to the liver). The metastatic cancer may include cells that exhibit migration and / or invasion of migratory cells, and / or cells that exhibit endothelial recruitment and / or angiogenesis. In other embodiments, the migratory cancer is a cell migration cancer. In yet other embodiments, the cell migration cancer is a non-metastatic cell migration cancer. The metastatic cancer may be a cancer that spreads by seeding on the surface of the peritoneum, pleura, pericardium, or subarachnoid space. Alternatively, the metastatic cancer may be a cancer that spreads through the lymphatic system, or a cancer that spreads through the bloodstream. In certain embodiments, the effective amount of the agent that reduces the level and / or activity of BRG1 and / or BRM is an amount that effectively inhibits metastatic colonization of the cancer to the liver.

[0481] In certain embodiments, the cancer contains a mutation in GNAQ. In certain embodiments, the cancer contains a mutation in GNA11. In certain embodiments, the cancer contains a mutation in PLCB4. In certain embodiments, the cancer contains a mutation in CYSLTR2. In certain embodiments, the cancer carries a mutation in BAP1. In certain embodiments, the cancer contains a mutation in SF3B1. In certain embodiments, the cancer contains a mutation in EIF1AX. In certain embodiments, the cancer contains a TFE3 translocation. In certain embodiments, the cancer contains a TFEB translocation. In certain embodiments, the cancer contains a MITF translocation. In certain embodiments, the cancer contains an EZH2 mutation. In certain embodiments, the cancer contains a SUZ12 mutation. In certain embodiments, the cancer contains an EED mutation.

[0482] In certain embodiments, the method further comprises administering to the subject an anti-cancer therapy or contacting the cell with an anti-cancer therapy, e.g., a chemotherapeutic or cytotoxic agent, immunotherapy, surgery, radiation therapy, thermotherapy, or photocoagulation. In certain embodiments, the anti-cancer therapy is a chemotherapeutic or cytotoxic agent, e.g., an antimetabolite, an antimitotic, an antitumor antibiotic, an asparagine-specific enzyme, a bisphosphonate, an antineoplastic agent, an alkylating agent, a DNA repair enzyme inhibitor, a histone deacetylase inhibitor, a corticosteroid, a demethylating agent, an immunomodulator, a janus-related kinase inhibitor, a phosphatidylinositol 3-kinase inhibitor, a proteasome inhibitor, or a tyrosine kinase inhibitor.

[0483] In certain embodiments, the compounds of the present invention are used in combination with another anticancer therapy for the treatment of uveal melanoma, such as surgery, a MEK inhibitor, and / or a PKC inhibitor. For example, in certain embodiments, the method further comprises performing surgery before, after, or simultaneously with the administration of the compound of the present invention. In certain embodiments, the method further comprises administering a MEK inhibitor and / or a PKC inhibitor before, after, or simultaneously with the administration of the compound of the present invention.

[0484] In certain embodiments, the anti-cancer therapy and the compound of the invention are administered within 28 days of each other, and each is administered in an amount that together is effective to treat the subject.

[0485] In certain embodiments, the subject or cancer has and / or has been identified as having a BRG1 loss-of-function mutation. In certain embodiments, the subject or cancer has and / or has been identified as having a BRM loss-of-function mutation.

[0486] In certain embodiments, the cancer is resistant to one or more chemotherapeutic or cytotoxic agents (e.g., the cancer has been identified as resistant to a chemotherapeutic or cytotoxic agent, such as by a genetic marker, or is likely to be resistant to a chemotherapeutic or cytotoxic agent, such as a cancer that has already failed to respond to a chemotherapeutic or cytotoxic agent). In certain embodiments, the cancer has already failed to respond to one or more chemotherapeutic or cytotoxic agents. In certain embodiments, the cancer is resistant to or has failed to respond to dacarbazine, temozolomide, cisplatin, treosulfan, fotemustine, IMCgp100, a CTLA-4 inhibitor (e.g., ipilimumab), a PD-1 inhibitor (e.g., nivolumab or pembrolizumab), a PD-L1 inhibitor (e.g., atezolizumab, avelumab, or durvalumab), a mitogen-activated protein kinase (MEK) inhibitor (e.g., selumetinib, binimetinib, or trametinib), and / or a protein kinase C (PKC) inhibitor (e.g., sotrastaurin or IDE196).

[0487] In certain embodiments, the cancer is resistant or unresponsive to a previously administered therapeutic agent for treating uveal melanoma, such as a MEK inhibitor or a PKC inhibitor. For example, in certain embodiments, the cancer is resistant or unresponsive to a mitogen-activated protein kinase (MEK) inhibitor (e.g., selumetinib, bimetinib, or trametinib) and / or a protein kinase C (PKC) inhibitor (e.g., sotrastaurin or IDE196).

[0488] Chemical terms

[0489] The terminology used herein is for the purpose of describing particular embodiments and is not intended to be limiting.

[0490] For any of the following chemical definitions, the number following the atomic symbol indicates the total number of atoms of that element present in the particular chemical moiety. As will be understood, other atoms (such as H atoms) or substituent groups as described herein may be present as needed to satisfy the valence of the atoms. For example, an unsubstituted C2 alkyl group has the formula -CH2CH3. When used with the groups defined herein, references to the number of carbon atoms include the divalent carbons in acetal and ketal groups, but do not include the carbonyl carbon in acyl, ester, carbonate, or carbamate groups. References to the number of oxygen, nitrogen, or sulfur atoms in a heteroaryl group include only those atoms that form part of the heterocycle.

[0491] As used herein, the term "acyl" represents an H or alkyl group attached to a parent molecular moiety via a carbonyl group as defined herein, and is exemplified by formyl (i.e., formaldehyde), acetyl, trifluoroacetyl, propionyl, and butyryl. Exemplary unsubstituted acyl groups include 1 to 6, 1 to 11, or 1 to 21 carbon atoms.

[0492] The term "alkyl" used herein refers to a branched or straight-chain monovalent saturated aliphatic hydrocarbon group of 1 to 20 carbon atoms (e.g., 1 to 16 carbon atoms, 1 to 10 carbon atoms, 1 to 6 carbon atoms, or 1 to 3 carbon atoms).

[0493] Alkylene is a divalent alkyl group. The term "alkenyl" as used herein, alone or in combination with other groups, refers to a straight or branched hydrocarbon group having one carbon-carbon double bond and 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).

[0494] The term "alkynyl," as used herein, alone or in combination with other groups, refers to a straight or branched chain hydrocarbon group having one 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).

[0495] The term "amino" as used herein represents -N(R N1 )2, where each R N1 are independently H, OH, NO2, N(R N2 2. SO2OR N2 、SO2R N2 、SOR N2 、 N - protecting group, alkyl, alkoxy, aryl, arylalkyl, cycloalkyl, acyl (eg, acetyl, trifluoroacetyl, or other acyl groups described herein), wherein these listed R N1 Each of the groups may be optionally substituted; or both R N1 to form an alkylene or heteroalkylene group, and wherein each R N2 are independently H, alkyl or aryl. The amino groups of the present invention can be unsubstituted amino (ie, -NH2) or substituted amino (ie, -N(R N1 )2).

[0496] As used herein, the term "aryl" refers to an aromatic mono- or polycyclic 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.

[0497] The term "arylalkyl" as used herein represents an alkyl group substituted with an aryl group. Exemplary unsubstituted arylalkyl groups have 7 to 30 carbon atoms (e.g., 7 to 16 or 7 to 20 carbon atoms, such as C1-C6 alkyl, C6-C 10 Aryl, C1-C 10 Alkyl C6-C 10 Aryl or C1-C 20 Alkyl C6-C 10 In certain embodiments, the alkyl and aryl groups may each be further substituted with 1, 2, 3, or 4 substituent groups as defined herein for each group.

[0498] The term "azido" as used herein represents a -N3 group.

[0499] As used herein, the term "bridged polycycloalkyl" refers to a bridged polycyclic group of 5 to 20 carbons containing 1 to 3 bridges.

[0500] The term "cyano" as used herein represents a -CN group.

[0501] The term "carbocyclyl" as used herein refers to a non-aromatic C3-C13-C16 ring formed by carbon atoms. 12 Monocyclic, bicyclic or tricyclic structures.Carbocyclic structures include cycloalkyl groups and unsaturated carbocyclic groups.

[0502] As used herein, the term "cycloalkyl" refers to a saturated, non-aromatic, monovalent, mono- or polycarbocyclic group of 3 to 10, preferably 3 to 6, carbon atoms. The term is further exemplified by groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, and adamantyl.

[0503] As used herein, the term "halogen" refers to a fluoro (F), chloro (Cl), bromo (Br), or iodo (Io) group.

[0504] As used herein, the term "heteroalkyl" refers to an alkyl group as defined herein, wherein one or more of the component carbon atoms have been replaced by nitrogen, oxygen or sulfur. In certain embodiments, the heteroalkyl group can be further replaced by 1, 2, 3 or 4 substituent groups as described herein for an alkyl group. The example of a heteroalkyl group is "alkoxy", which refers to alkyl-O- (e.g., methoxy and ethoxy) as used herein. Heteroalkylene is a divalent heteroalkyl group. As used herein, the term "heteroalkenyl" refers to an alkenyl group as defined herein, wherein one or more of the component carbon atoms have been replaced by nitrogen, oxygen or sulfur. In certain embodiments, the heteroalkenyl group can be further replaced by 1, 2, 3 or 4 substituent groups as described herein for an alkenyl group. The example of a heteroalkenyl group is "alkenyloxy", which refers to alkenyl-O- as used herein. Heteroalkenylene is a divalent heteroalkenyl group. As used herein, the term "heteroalkynyl" refers to an alkynyl group as defined herein, wherein one or more of the component carbon atoms have been replaced by nitrogen, oxygen or sulfur. In certain embodiments, the heteroalkynyl group can be further substituted with 1, 2, 3, or 4 substituent groups as described herein for alkynyl groups. An example of a heteroalkynyl group is "alkynyloxy," which as used herein refers to alkynyl-O-. Heteroalkynylene is a divalent heteroalkynyl group.

[0505] The term "heteroaryl" as used herein refers to a monocyclic or polycyclic group of 5 to 12 atoms having at least one aromatic ring containing 1, 2 or 3 ring atoms selected from nitrogen, oxygen and sulphur, and the remaining ring atoms are carbon. One or two ring carbon atoms of the heteroaryl group can be replaced by a carbonyl group. Examples of heteroaryl groups are pyridyl, pyrazolyl, benzoxazolyl, benzimidazolyl, benzothiazolyl, imidazolyl, oxaxolyl and thiazolyl.

[0506] The term "heteroarylalkyl" as used herein represents an alkyl group substituted with a heteroaryl group. Exemplary unsubstituted heteroarylalkyl groups have 7 to 30 carbon atoms (e.g., 7 to 16 or 7 to 20 carbon atoms, such as C1-C6 alkyl, C2-C9 heteroaryl, C1-C 10 Alkyl C2-C9 heteroaryl or C1-C 20 In certain embodiments, the alkyl and heteroaryl groups may each be further substituted with 1, 2, 3, or 4 substituent groups as defined herein for each group.

[0507] As used herein, the term "heterocyclyl" refers to a monocyclic or polycyclic group having from 3 to 12 atoms, having at least one ring containing 1, 2, 3, or 4 ring atoms selected from N, O, or S, wherein no ring is aromatic. Examples of heterocyclyl groups include, but are not limited to, morpholinyl, thiomorpholinyl, furanyl, piperazinyl, piperidinyl, pyranyl, pyrrolidinyl, tetrahydropyranyl, tetrahydrofuranyl, and 1,3-dioxanyl.

[0508] The term "heterocyclylalkyl" as used herein represents an alkyl group substituted by a heterocyclyl group. Exemplary unsubstituted heterocyclylalkyl groups have 7 to 30 carbon atoms (e.g., 7 to 16 or 7 to 20 carbon atoms, such as C1-C6 alkyl, C2-C9 heterocyclyl, C1-C 10 Alkyl C2-C9 heterocyclic group or C1-C 20 In certain embodiments, the alkyl and heterocyclyl groups may each be further substituted with 1, 2, 3, or 4 substituent groups as defined herein for each group.

[0509] The term "hydroxyalkyl" as used herein represents an alkyl group substituted with an -OH group.

[0510] As used herein, the term "hydroxy" represents an -OH group.

[0511] The term " N "-Protective groups" represent those groups intended to protect amino groups against undesired reactions during synthetic procedures. Commonly used N - Protecting groups are disclosed in Greene, "Protective Groups in Organic Synthesis," 3rd edition (John Wiley & Sons, New York, 1999). N- Protecting groups include, but are not limited to, acyl, aroyl, or carbamoyl groups such as formyl, acetyl, propionyl, pivaloyl, tert-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthaloyl, o-nitrophenoxyacetyl, α-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, 4-nitrobenzoyl; and chiral auxiliaries such as protected or unprotected D, L, or D, L-amino acids such as alanine, leucine, and phenylalanine; sulfonyl-containing groups such as benzylsulfonyl and p-toluenesulfonyl; carbamate-forming groups such as benzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl, 2,4-20-dimethoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl, 3,4,5-trimethoxybenzyloxycarbonyl, 1-(p- phenyl)-1-methylethoxycarbonyl, α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, benzhydryloxycarbonyl, tert-butyloxycarbonyl, diisopropylmethoxycarbonyl, isopropyloxycarbonyl, ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl, 2,2,2,-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl, cyclopentyloxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl and phenylthiocarbonyl; arylalkyl groups such as benzyl, triphenylmethyl and benzyloxymethyl; and silyl groups such as trimethylsilyl. Preferred N - Protecting groups are alloc, formyl, acetyl, benzoyl, pivaloyl, tert-butylacetyl, alanyl, phenylsulfonyl, benzyl, tert-butyloxycarbonyl (Boc) and benzyloxycarbonyl (Cbz).

[0512] The term "nitro" as used herein represents a -NO2 group.

[0513] The term "mercapto" as used herein represents a -SH group.

[0514] The alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl (e.g., cycloalkyl), aryl, heteroaryl and heterocyclyl groups can be substituted or unsubstituted. When substituted, there will generally be 1 to 4 substituents, unless otherwise noted. Substituents include, for example, alkyl (e.g., unsubstituted and substituted, wherein the substituent includes any group described herein, e.g., aryl, halogen, hydroxyl), aryl (e.g., substituted and unsubstituted phenyl), carbocyclyl (e.g., substituted and unsubstituted cycloalkyl), halogen (e.g., fluorine), hydroxyl, heteroalkyl (e.g., substituted and unsubstituted methoxy, ethoxy or thioalkoxy), heteroaryl, heterocyclyl, amino (e.g., NH or mono- or dialkylamino), azido, cyano, nitro, oxo or sulfhydryl. In certain embodiments, substituents include: alkyl (e.g., unsubstituted and substituted, wherein the substituents include any of the groups described herein, e.g., aryl, halogen, hydroxy), aryl (e.g., substituted and unsubstituted phenyl), carbocyclyl (e.g., substituted and unsubstituted cycloalkyl), halogen (e.g., fluoro), hydroxy, heteroalkyl (e.g., substituted and unsubstituted methoxy, ethoxy, or thioalkoxy), heteroaryl, heterocyclyl, amino (e.g., NH or mono- or dialkylamino), azido, cyano, nitro, or thiol. Aryl, carbocyclyl (e.g., cycloalkyl), heteroaryl, and heterocyclyl groups can also be substituted with alkyl (unsubstituted and substituted, such as arylalkyl (e.g., substituted and unsubstituted benzyl)).

[0515] The compounds of the present invention may have one or more asymmetric carbon atoms and may exist in the following forms: optically pure enantiomers, mixtures of enantiomers such as, for example, racemates, optically pure diastereomers, mixtures of diastereomers, diastereomeric racemates, or mixtures of diastereomeric racemates. Optically active forms can be obtained, for example, by resolving racemates, by asymmetric synthesis, or by asymmetric chromatography (chromatography using a chiral adsorbent or eluent). That is, certain disclosed compounds may exist in various stereoisomeric forms. Stereoisomers are compounds that differ only in their spatial arrangement. Enantiomers are paired stereoisomers whose mirror images are not superimposable, most commonly because they contain an asymmetrically substituted carbon atom that serves as a chiral center. "Enantiomer" means one of a pair of molecules that are mirror images of each other and are not superimposable. Diastereoisomers are stereoisomers that are not related to 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. Enantiomers of a compound can be prepared, for example, by separating one enantiomer from a racemate using one or more well-known techniques and methods, such as, for example, chiral chromatography and separation methods based thereon. Those skilled in the art can readily determine appropriate techniques and / or methods for separating enantiomers of the compounds described herein from a racemic mixture. "Racemate" or "racemic mixture" refers to a compound containing two enantiomers, wherein such a mixture exhibits no optical activity; that is, they do not rotate the plane of polarized light. "Geometric isomer" refers to isomers that differ in the orientation of substituent atoms relative to a carbon-carbon double bond, a cycloalkyl ring, or a bridged bicyclic ring system. Atoms on each side of a carbon-carbon double bond (except H) can be in the E (substituents are on opposite sides of the carbon-carbon double bond) or Z (substituents are oriented on the same side) configuration. "R," "S," "S*," "R*," "E," "Z," "cis," and "trans" indicate configuration relative to the core molecule. Certain disclosed compounds may exist in atropisomeric forms. Atropisomers are stereoisomers resulting from hindered rotation about a single bond, where the steric barrier to rotation is high enough to allow separation of the conformers. The compounds of the present invention can be prepared as single isomers by isomer-specific synthesis or resolution from an isomeric mixture. Conventional resolution techniques include forming salts of the free bases of each isomer of the isomeric 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 the isomeric pair using an optically active amine (followed by fractional crystallization and regeneration of the free acid), forming esters or amides of each isomer of the isomeric pair using an optically pure acid, amine, or alcohol (followed by chromatographic separation and removal of the chiral auxiliary), or resolving isomeric mixtures of the starting material or final product using various well-known chromatographic methods.When the stereochemistry of a disclosed compound is named or depicted by structure, the stereoisomer named or depicted is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by weight relative to the other stereoisomers. When a single enantiomer is named or depicted by structure, the enantiomer depicted or named is at least 60%, 70%, 80%, 90%, 99%, or 99.9% optically pure by weight. When a single diastereomer is named or depicted by structure, the diastereomer depicted or named is at least 60%, 70%, 80%, 90%, 99%, or 99.9% pure by weight. Percent optical purity is the ratio of the weight of the enantiomer to the weight of the enantiomer plus the weight of its optical isomers. Diastereomeric purity by weight is the ratio of the weight of one diastereomer to the weight of all diastereomers. When the stereochemistry of a disclosed compound is named or depicted by structure, the stereoisomer named or depicted has a purity of at least 60%, 70%, 80%, 90%, 99%, or 99.9% by mole relative to the other stereoisomers. When a single enantiomer is named or depicted by structure, the enantiomer described or named has a purity of at least 60%, 70%, 80%, 90%, 99%, or 99.9% by mole. When a single diastereomer is named or depicted by structure, the diastereomer described or named has a purity of at least 60%, 70%, 80%, 90%, 99%, or 99.9% by mole. The percent purity by mole is the ratio of the moles of enantiomer to the moles of enantiomer plus the moles of its optical isomers. Similarly, the percent purity by mole is the ratio of the moles of diastereomer to the moles of diastereomer plus the moles of its isomers. When a disclosed compound is named or depicted by structure without indicating stereochemistry and the compound has at least one chiral center, it is understood that the name or structure encompasses the enantiomers of the compound free of the corresponding optical isomers, the racemic mixture of the compound, or a mixture enriched in one enantiomer relative to its corresponding optical isomer. When a disclosed compound is named or depicted by structure without indicating stereochemistry and has two or more chiral centers, it is understood that the name or structure encompasses the diastereomer free of the other diastereomers, a plurality of diastereomers free of other diastereomeric pairs, a mixture of diastereomers, a mixture of diastereomeric pairs, a mixture of diastereomers in which one diastereomer is enriched relative to the other diastereomers, or a mixture of diastereomers in which one or more diastereomers are enriched relative to the other diastereomers. The present invention includes all of these forms.

[0516] Compounds of the present disclosure also include all isotopes of atoms present in intermediate compounds or final compounds. "Isotopes" refer to atoms having the same atomic number but different mass numbers resulting from different numbers of neutrons in the nucleus. For example, isotopes of hydrogen include tritium and deuterium.

[0517] Unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. Exemplary isotopes that can be incorporated into 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. Isotope-labeled compounds (e.g., 3 H and 14 C-labeled) can be used in compound or substrate tissue distribution assays. Because of their ease of preparation and detectability, tritiated (i.e., 3 H) and carbon-14 (i.e., 14 C) isotopes can be useful. In addition, heavier isotopes such as deuterium (i.e., 2 ) substitution may provide certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements). In certain embodiments, one or more hydrogen atoms are replaced by 2 H or 3 H is replaced, or one or more carbon atoms are replaced 13 C- or 14 C-enriched carbon substitutes. Positron-emitting isotopes such as 15 O. 13 N. 11 C and 18F can be used in positron emission tomography (PET) studies to examine substrate receptor occupancy. The preparation of isotope-labeled compounds is known to those skilled in the art. For example, isotope-labeled compounds can generally be prepared by replacing non-isotope-labeled reagents with isotope-labeled reagents according to procedures similar to those disclosed for the compounds of the present invention described herein. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the invention belongs. The methods and materials used in the present disclosure are described herein; other suitable methods and materials known in the art can also be used. The materials, methods, and examples are exemplary only and are not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In the event of a conflict, the present specification (including definitions) shall prevail.

[0518] definition

[0519] In this application, unless otherwise clear from the context, (i) the term "a" may be understood to mean "at least one"; (ii) the term "or" may be understood to mean "and / or"; and (iii) the terms "comprising" and "including" may be understood to cover the itemized components or steps, whether presented alone or together with one or more additional components or steps.

[0520] As used herein, the terms "about" and "approximately" refer to values ​​that are within 10% above or below the value being described. For example, the term "about 5 nM" indicates a range of 4.5 to 5.5 nM.

[0521] As used herein, the term "administering" refers to administering a composition (e.g., a compound or a preparation comprising a compound described herein) to a subject or system. Administration to an animal subject (e.g., to a human) can be by any appropriate route. For example, in certain embodiments, administration can be intrabronchial (including by bronchial instillation), buccal, enteral, intradermal, intraarterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intratumoral, intravenous, intraventricular, mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (including by intratracheal instillation), transdermal, vaginal, and vitreous.

[0522] As used herein, the term "BAF complex" refers to a BRG1- or HRBM-related factor complex in human cells.

[0523] As used herein, the term "BAF complex-associated disorder" refers to a disorder that is caused or affected by the activity level of the BAF complex.

[0524] As used herein, the term "BRG1 loss-of-function mutation" refers to a mutation in BRG1 that results in the protein having reduced 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.

[0525] As used herein, the term "BRG1 loss-of-function disorder" refers to a disorder (e.g., cancer) exhibiting reduced BRG1 activity (e.g., at least 1% reduction in BRG1 activity, such as 2%, 5%, 10%, 25%, 50%, or 100% reduction in BRG1 activity).

[0526] The term "cancer" refers to disorders caused by the proliferation of malignant neoplastic cells, such as tumors, neoplasms, carcinomas, sarcomas, leukemias, and lymphomas.

[0527] As used herein, "combination therapy" or "combination administration" means administering two (or more) different agents or treatments to a subject as part of a defined treatment regimen for a specific disease or condition. The treatment regimen defines the dosage and duration of administration of each agent so that the effects of the various agents on the subject overlap. In certain embodiments, the two or more agents are delivered simultaneously or concurrently, and the agents may be co-formulated. In certain embodiments, the two or more agents are not co-formulated and are administered sequentially as part of a prescribed regimen. In certain embodiments, the combined administration of two or more agents or treatments results in a reduction in symptoms or other parameters associated with the disorder that is greater than that observed when either agent or treatment is delivered alone, or in the absence of the other agent or treatment. The effects of the two treatments may be partially additive, fully additive, or greater than additive (e.g., synergistic). Sequential or substantially simultaneous administration of each therapeutic agent can be achieved by any appropriate route, including, but not limited to, oral, intravenous, intramuscular, and direct absorption through mucosal tissues. The therapeutic agents may be administered by the same route or by different routes. For example, a first therapeutic agent of the combination may be administered by intravenous injection, while a second therapeutic agent of the combination may be administered orally.

[0528] "Determining the level of a protein or RNA" means detecting a protein or RNA directly or indirectly by methods known in the art. "Directly determining" means performing a process (e.g., performing an assay or test on a sample, or "analyzing a sample," as the term is defined herein) to obtain a physical entity or value. "Indirectly determining" refers to receiving a physical entity or value from another party or source (e.g., a third-party laboratory that directly obtains the physical entity or value). Methods for measuring protein levels generally include, but are not limited to, western blotting, immunoblotting, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunoprecipitation, immunofluorescence, surface plasmon resonance, chemiluminescence, fluorescence polarization, phosphorescence, immunohistochemical analysis, matrix-assisted laser desorption / ionization time-of-flight (MALDI-TOF) mass spectrometry, liquid chromatography (LC)-mass spectrometry, microcytometry, microscopy, fluorescence-activated cell sorting (FACS) and flow cytometry, as well as assays based on protein properties (including, but not limited to, enzyme activity or interactions with other protein partners). Methods of measuring RNA levels are known in the art and include, but are not limited to, quantitative polymerase chain reaction (qPCR) and Northern blot analysis.

[0529] "Reducing the activity of the BAF complex" means reducing the level of activity associated with the BAF complex or a related downstream effect. A non-limiting example of reducing the activity of the BAF complex is Sox2 activation. The level of activity of the BAF complex can be measured using any method known in the art, for example, the method described in Kadoch et al. Cell, 2013, 153, 71-85, which is incorporated herein by reference.

[0530] As used herein, the term "degrader" refers to a small molecule compound that includes a degrading moiety, wherein the compound interacts with a protein (e.g., BRG1 and / or BRM) in a manner that results in degradation of the protein, e.g., binding of the compound results in at least a 5% decrease in protein levels, e.g., in a cell or subject.

[0531] As used herein, the term "degradation moiety" refers to a moiety whose binding results in degradation of a protein (e.g., BRG1 and / or BRM). In one embodiment, the moiety binds to a protease or ubiquitin ligase that metabolizes the protein (e.g., BRG1 and / or BRM).

[0532] "Modulating the activity of a BAF complex" means changing the level of activity associated with a BAF complex (e.g., GBAF) or an associated downstream effect. The level of activity of a BAF complex can be measured using any method known in the art, for example, as described in Kadoch et al. Cell, 2013, 153, 71-85, which is incorporated herein by reference.

[0533] "Reducing the activity of BRG1 and / or BRM" means reducing the level of activity associated with BRG1 and / or BRM or a related downstream effect. A non-limiting example of inhibiting the activity of BRG1 and / or BRM is 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 certain embodiments, the agent that reduces the activity of BRG1 and / or BRM is a small molecule BRG1 and / or BRM degrader.

[0534] "Decreasing the level of BRG1 and / or BRM" means 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.

[0535] "Level" refers to the level of a protein or mRNA encoding a protein compared to a reference. The reference can be any useful reference as defined herein. A "reduced level" or "increased level" of a protein means a decrease or increase in protein levels compared to a 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 30%, about 400%, about 500% or more compared to a reference). The level of a protein can be expressed as a decrease or increase of about 15%, about 20%, about 50%, about 75%, about 100%, or about 200%; 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 mRNA in the sample.

[0536] As used herein, the term "inhibit BRM" refers to blocking or reducing the level or activity of the ATPase catalytic binding domain or bromodomain of a protein. BRM inhibition can be determined using methods known in the art, such as a BRM ATPase assay, a Nano DSF assay, or a BRM luciferase cell assay.

[0537] As used herein, the term "pharmaceutical composition" refers to a composition containing a compound described herein, formulated with a pharmaceutically acceptable excipient and suitable for administration to a mammal, such as a human. Typically, a pharmaceutical composition is manufactured or sold with the approval of a governmental regulatory agency as part of a therapeutic regimen for treating a disease in a mammal. Pharmaceutical compositions can be formulated, for example, for oral administration in a unit dosage form (e.g., tablets, capsules, caplets, gel caps, or syrups); for topical administration (e.g., as a cream, gel, lotion, or ointment); for intravenous administration (e.g., as a sterile solution without microparticles and in a solvent system suitable for intravenous use); or in any other pharmaceutically acceptable formulation.

[0538] As used herein, a "pharmaceutically acceptable excipient" refers to any ingredient other than a compound described herein (e.g., a vehicle capable of suspending or dissolving the active compound) that is substantially non-toxic and non-inflammatory in a patient. Excipients may include, for example, anti-adherents, antioxidants, binders, coating agents, compression aids, disintegrants, dyes (colorants), softeners, emulsifiers, fillers (diluents), film-forming or coating agents, flavorings, fragrances, glidants (flow promoters), lubricants, preservatives, printing inks, sorbents, suspending or dispersing agents, sweeteners, and water of hydration.

[0539] As used herein, the term "pharmaceutically acceptable salt" means any pharmaceutically acceptable salt of a compound (e.g., any compound of Formula I or II). Pharmaceutically acceptable salts of any compound described herein may include salts that are suitable for use in contact with human and animal tissues, without undue toxicity, irritation, or allergic response, and commensurate with a reasonable benefit / risk ratio, within the scope of reasonable medical judgment. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in Berge et al., J. Pharmaceutical Sciences 66: 1-19, 1977 and Pharmaceutical Salts: Properties, Selection, and Use, (Eds. P.H. Stahl and C.G. Wermuth), Wiley-VCH, 2008. Salts can be prepared in situ during the final isolation and purification of the compounds described herein, or separately by reacting the free base group with a suitable organic acid.

[0540] The compounds of the present invention may have ionizable groups so that they can be prepared into pharmaceutically acceptable salts. These salts may be acid addition salts involving inorganic or organic acids, or in the case of acidic forms of the compounds of the present invention, the salts may be prepared from inorganic or organic bases. Frequently, the compounds are prepared or used as pharmaceutically acceptable salts, which are prepared as addition products of pharmaceutically acceptable acids or bases. Suitable pharmaceutically acceptable acids and bases and methods for preparing appropriate salts are well known in the art. Salts may be prepared from pharmaceutically acceptable non-toxic acids and bases, including inorganic and organic acids and bases.

[0541] "Reference" means any useful reference for comparing protein or RNA levels. A reference can be any sample, standard, standard curve, or level used for comparison purposes. A reference can be a normal reference sample or reference standard or level. A "reference sample" can be, for example, a control, such as a predetermined negative control value, such as a "normal control" or a previous sample taken from the same subject; a sample from a normal, healthy subject, such as normal cells or normal tissue; a sample (e.g., cells or tissue) from a subject without disease; a sample from a subject diagnosed with a disease but not yet treated with a compound of the invention; a sample from a subject treated with a compound of the 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 predetermined value indicative of a non-disease state, e.g., a value expected in a healthy control subject. Typically, normal control values ​​are expressed as a range ("between X and Y"), an upper threshold ("not higher than X"), or a lower threshold ("not lower than X"). A subject with a measured value for a particular biomarker that falls within a normal control value is typically said to be "within normal limits" for that biomarker. A normal reference standard or level can be a value or number derived from: a normal subject without a disease or disorder (e.g., cancer); or a subject who has been treated with a compound of the invention. In preferred embodiments, the reference sample, standard, or level is matched to the sample subject by at least one of the following criteria: age, weight, sex, disease stage, and overall health. A standard curve of levels of purified protein or RNA (e.g., any of those described herein) within the normal reference range can also be used as a reference.

[0542] As used herein, the term "subject" refers to any organism to which a composition according to the present invention can be administered, e.g., 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 may be seeking or in need of treatment, requiring treatment, currently receiving treatment, will be receiving treatment in the future, or may be a human or animal under the care of a trained professional for a particular disease or condition.

[0543] As used herein, the terms "treat," "treated," or "treating" refer to therapeutic treatment or any measure whose purpose is to slow down (lessen) an undesirable physiological condition, disorder, or disease, or to achieve a beneficial or desired clinical result. Beneficial or desired clinical results include, but are not limited to: alleviation of symptoms; reduction in the extent of the condition, disorder, or disease; stabilization of the condition, disorder, or disease (i.e., no worsening); delay in the onset or slowing of the progression of the condition, disorder, or disease; amelioration or remission (whether partial or complete) of the condition, disorder, or disease state; improvement in at least one measurable physical parameter, not necessarily discernible by the patient; or amelioration or improvement of the condition, disorder, or disease. Treatment includes eliciting a clinically significant response without undue side effects. Treatment also includes prolonging survival as compared to expected survival if not receiving treatment. The compounds of the present invention may also be used to "prophylactically treat" or "prevent" a disorder, for example, in a subject at increased risk of developing the disorder.

[0544] As used herein, the terms "variant" and "derivative" are used interchangeably and refer to naturally occurring, synthetic, and semisynthetic analogs of the compounds, peptides, proteins, or other substances described herein. Variants or derivatives of compounds, peptides, proteins, or other substances described herein may retain or improve the biological activity of the original material.

[0545] The details of one or more embodiments of the invention are set forth in the description which follows. Other features, objects, and advantages of the invention will be apparent from the description and from the claims.

[0546] Detailed description

[0547] The present disclosure features compounds that can be used to inhibit BRG1 and / or BRM. These compounds can be used to modulate the activity of the BAF complex, for example, for treating BAF-related disorders such as cancer. Exemplary compounds described herein include compounds having a structure according to Formula I or II, or pharmaceutically acceptable salts thereof.

[0548] Formula I:

[0549]

[0550] in

[0551] X 1 Is O or NR 2 ;

[0552] Each X 2 are independently halogen;

[0553] k is 0, 1, 2, 3, or 4;

[0554] m is 0, 1, 2, 3, or 4;

[0555] R 1 is halogen or optionally substituted C1-C6 alkyl;

[0556] R 2 is H or optionally substituted C1-C6 alkyl;

[0557] L 1 is an optionally substituted C1-C6 alkylene group;

[0558] L is included A linker of a structure;

[0559] n is 0, 1, 2, or 3;

[0560] L 2 is an optionally substituted C1-C6 alkylene, an optionally substituted C1-C 20 heteroalkylene or optionally substituted C2-C9 heterocyclylene;

[0561] Each L 3 are independently -O-, optionally substituted C1-C 20 Heteroalkylene, optionally substituted C3-C 10 Carbocyclylene, optionally substituted C3-C 10 Carbocyclylene-C1-C 20 an alkylene group, an optionally substituted C2-C9 heterocyclylene group, or an optionally substituted C2-C9 heterocyclylene group-C1-C 20 an alkylene group; and

[0562] D is the degradation fraction.

[0563] Formula II:

[0564]

[0565] in

[0566] A Z 1 and a Z 2 to form an optionally substituted C1-C4 alkylene group, and the remaining Z 1 and Z 2 Each is hydrogen;

[0567] Each X 2 are independently halogen;

[0568] k is 0, 1, 2, 3, or 4;

[0569] L is A linker of a structure;

[0570] q is 0, 1, 2, 3, or 4;

[0571] L 4 is an optionally substituted C1-C6 alkylene, an optionally substituted C1-C 20 heteroalkylene or optionally substituted C2-C9 heteroarylene;

[0572] Each L 5 are independently -O-, optionally substituted C1-C6 alkylene, optionally substituted C1-C 20 Heteroalkylene, optionally substituted C3-C 10 Carbocyclylene, optionally substituted C3-C 10 Carbocyclylene-C1-C6 alkylene, optionally substituted C2-C9 heterocyclylene or C2-C9 heterocyclylene-C1-C 20 an alkylene group; and

[0573] D is the degradation fraction.

[0574] In certain embodiments, the compound has the structure of any one of Compounds 1-75 in Table 1 or Compounds 76-104 in Table 2, or a pharmaceutically acceptable salt thereof.

[0575] Other embodiments are described herein, as well as exemplary methods for synthesizing or producing these compounds.

[0576] Drug uses

[0577] The compounds described herein can be used in the methods of the present invention, and while not being bound by theory, it is believed that they exert their ability to modulate the level, status, and / or activity of the BAF complex by inhibiting the activity of BRG1 and / or BRM proteins within the BAF complex in an active mammal. Disorders associated with the BAF complex include, but are not limited to, disorders associated with loss-of-function mutations in BRG1.

[0578] One aspect of the invention relates to methods for treating a disorder associated with a BRG1 loss-of-function mutation, such as cancer (e.g., non-small cell lung cancer, colorectal cancer, bladder cancer, cancer of unknown primary, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, or penile cancer) in a subject in need thereof. In certain embodiments, the compound is administered in an amount and for a time effective to result in one or more (e.g., two or more, three or more, four or more) of the following: (a) decreased tumor size, (b) decreased tumor growth rate, (c) increased tumor cell death, (d) decreased tumor progression, (e) decreased number of metastases, (f) decreased metastasis rate, (g) decreased tumor recurrence, (h) increased subject survival, (i) increased subject progression-free survival.

[0579] Treating cancer can result in a decrease in the size or volume of a tumor. For example, after treatment, the size of a tumor decreases by 5% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more) relative to its size before treatment. Tumor size can be measured by any reproducible measurement method. For example, tumor size can be measured as the diameter of the tumor.

[0580] Treatment of cancer can further result in a decrease in the number of tumors. 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 method. For example, the number of tumors can be measured by counting tumors visible to the naked eye or at a specific magnification (e.g., 2x, 3x, 4x, 5x, 10x, or 50x).

[0581] Treatment of cancer can result in a reduction in the number of metastatic nodules in other tissues or organs away from the primary tumor site. For example, after treatment, the number of metastatic nodules is reduced by 5% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more) relative to the number before treatment. The number of metastatic nodules can be measured by any reproducible measurement method. For example, the number of metastatic nodules can be measured by counting macroscopic metastatic nodules or at a specific magnification (e.g., 2x, 10x, or 50x).

[0582] Compared to an untreated subject population, treating cancer can result in an increase in the mean survival time of a subject population treated according to the present invention. For example, the mean survival time increases by more than 30 days (more than 60 days, 90 days, or 120 days). By any reproducible means, the increase in the mean survival time of a population can be measured. For example, by calculating the mean survival time after treatment with a compound of the present invention is started for a population, the increase in the mean survival time of a population can be measured. For example, by calculating the mean survival time after the first round of treatment with a pharmaceutically acceptable salt of the present invention is completed for a population, the increase in the mean survival time of a population can also be measured.

[0583] Treatment of cancer can also result in a reduction in the mortality rate of the treated subject population compared to an untreated population. For example, the mortality rate is reduced by more than 2% (e.g., more than 5%, 10%, or 25%). The reduction in the mortality rate of the treated subject population can be measured in any reproducible manner, for example, by calculating for the population the average number of disease-related deaths per unit time after starting treatment with a pharmaceutically acceptable salt of the present invention. For example, the reduction in population mortality can also be measured by calculating for the population the average number of disease-related deaths per unit time after completing the first round of treatment with a pharmaceutically acceptable salt of the present invention.

[0584] Exemplary cancers that can be treated by the present invention include, but are not limited to, non-small cell lung cancer, small cell lung cancer, colorectal cancer, bladder cancer, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, esophageal-gastric cancer, pancreatic cancer, hepatobiliary cancer, soft tissue sarcoma, ovarian cancer, head and neck cancer, renal cell carcinoma, bone cancer, non-Hodgkin lymphoma, prostate cancer, embryonal tumors, germ cell tumors, cervical cancer, thyroid cancer, salivary gland cancer, gastrointestinal neuroendocrine tumors, uterine sarcoma, gastrointestinal stromal tumors, central nervous system cancers, thymic tumors, adrenocortical carcinoma, appendix cancer, small intestine cancer, and penile cancer.

[0585] Combination preparations and their uses

[0586] The compounds of the present invention may be combined with one or more therapeutic agents. Specifically, the therapeutic agent may be a therapeutic agent for the treatment or prophylactic treatment of any of the cancers described herein.

[0587] Combination therapy

[0588] The compounds of the present invention can be used alone or in combination with another therapeutic agent (e.g., other agents for treating cancer or symptoms associated therewith), or in combination with other types of treatments for cancer. In combination therapy, the dose of one or more therapeutic compounds can be reduced from the standard dose when administered alone. For example, the dose can be determined empirically from the drug combination and arrangement, or the dose can be inferred by isobologram analysis (e.g., Black et al., Neurology 65: S3-S6, 2005). In this case, the dose of the compound should provide a therapeutic effect when combined.

[0589] In certain embodiments, the second therapeutic agent is a chemotherapeutic agent (e.g., a cytotoxic agent or other chemical compound useful for 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 suppressants, adrenocortical steroids, progestogens, estrogens, antiestrogens, androgens, antiandrogens, and gonadotropin-releasing hormone analogs. Also included are 5-fluorouracil (5-FU), folinic acid (LV), irinotecan, oxaliplatin, capecitabine, paclitaxel, and docetaxel. Non-limiting examples of chemotherapeutic agents include: alkylating agents such as thiotepa and cyclophosphamide; alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodepa, carboquinone, metodepa, and uredepa; ethyleneimines and methylmelamines, including hexamethylmelamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trishydroxymethylmelamine; annonaceous lactones (particularly bratacin and bratacinone); camptothecins (including the synthetic analogue topotecan); bryostatin; callystatin; CC-1065 (including its synthetic analogs adolesin, kazelesin and bitrazesin); Nostoc cyclic peptides (especially Nostoc cyclic peptide 1 and Nostoc cyclic peptide 8); dolastatin; duocarmycin (including synthetic analogs, KW-2189 and CB1-TM1); soft coral alcohol; hyoscyamine; sarcodictyin; spongestatin; nitrogen mustards such as chlorambucil, naphthyl mustard, clofosamide, estramustine, ifosfamide, dichloromethyldiethylamine, dichloromethyldiethylamine oxide hydrochloride, melphalan, new nitrogen mustard, phenylephrine, prednimustine, trofosamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine and ranimustine; antibiotics such as enediyne antibiotics (e.g., calicheamicin, especially calicheamicin γll and calicheamicin ωll (See, e.g., Agnew, Chem. Intl. Ed Engl.33:183-186 (1994)); daptomycins, including daptomycin A; bisphosphonates such as clodronate; esperamicins; and the neocarcin chromophores and related chromoprotein enediyne antibiotic chromophores), aclarubicin, actinomycin, anthramycin, azaserine, bleomycin, actinomycin C, carabicin, carminomycin, carmomycin, chromomycin, dactinomycin, daunorubicin, detoximcin, 6-diazo- 5-oxo-L-norleucine, Adriamycin® (doxorubicin, including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, mexicomycin, mitomycins such as mitomycin C, mycophenolic acid, noramycin, olivomycin, peplomycin, porfiromycin, puromycin, triferric doxorubicin, rhodorubicin, streptozocin, streptozocin, tuberculin, ubenimex, zoloft, daunorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5- FU); folic acid analogs such as dimethylformamide, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiopurine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as captestosterone, drostanolone propionate, cyclothiosteroid, melastane, testolactone; antiadrenal agents such as aminoglutethimide, mitotane, trilostane; folic acid supplements such as folinic acid; aceglucuronolide; aldophosphamide glycosides; aminoleucovorin acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatrexate; defosfamide; colcemid; diacronitrone; eflornithine; elliptonium acetate; epothilone; etoglucagon; gallium nitrate; hydroxyurea; lentinan; lonidamine; maytansines such as maytansine and ansamitocin; mitoxantrone; mopidanmol; nitraerine; pentostatin; methamidine; pirarubicin; losoxantrone; podophyllic acid; 2-ethylhydrazide; procarbazine; PSK® polysaccharide complex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin; sizofuran; spirogermanium; tenuzolic acid; triimidoquinone; 2,2',2''-trichlorotriethylamine; trichothecenes (particularly T-2 toxin, verrucosporin A, baculosporin A, and serpentin); urethan; vindesine; dacarbazine; mannomustine; dibromomannitol; dibromodulcitol; pipobroman; gacytosine; cytarabine ("Ara-C"); cyclophosphamide; thiotepa; taxanes, for example, Taxol® paclitaxel (Bristol-Myers Squibb Oncology, Princeton, NJ), Abraxane®, a cremophor-free, albumin-engineered nanoparticle formulation of paclitaxel (American Pharmaceutical Partners, Schaumberg, Ill.), and Taxotere® docetaxel (Rhone-Poulenc Rorer, Antony, France); chlorambucil; gemcitabine®; 6-thioguanine; mercaptopurine; methotrexate; platinum coordination complexes such as cisplatin, oxaliplatin, and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; vinorelbine®; nivolumab; teniposide; edatrexate; daunorubicin; aminopterin; xeloda; ibandronate; irinotecan (e.g., CPT-11); the 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 can be used in a mixture to be administered in combination with the first therapeutic agent described herein. Suitable dosing regimens for combination chemotherapy are known in the art and described, for example, in Saltz et al. (1999) Proc ASCO 18:233a and Douillard et al. (2000) Lancet 355:1041-7.

[0590] In certain embodiments, the second therapeutic agent is a therapeutic agent that is a biologic, such as a cytokine used in cancer treatment (e.g., an interferon or interleukin (e.g., IL-2)). In certain embodiments, the biologic is an anti-angiogenic agent, such as an anti-VEGF agent, for example, bevacizumab (Avastin®). In certain embodiments, the biologic is an immunoglobulin-based biologic, for example, a monoclonal antibody (e.g., a humanized antibody, a fully human antibody, an Fc fusion protein, or a functional fragment thereof) that agonizes a target to stimulate an anti-cancer response or antagonizes an antigen important for cancer. Such agents include Rituxan (rituximab); Zenapax (daclizumab); Sulai (basiliximab); Synagis (palivizumab); Remicade (infliximab); Herceptin (trastuzumab); Mylotarg (gemtuzumab ozogamicin); Campath (alemtuzumab); Zevalin (ibritumomab tiuxetan); Humira (adalimumab); Xolair (omalizumab); Bexxar (tositumomab-I-131); Raptiva (efalizumab); Erbitux (cetuximab); Avastin (bevacizumab); Tysabri (natalizumab); Actemra (tocilizumab); Vectibix (panitumumab); Lucentis (ranibizumab); Soliris (eculizumab); Cimzia (certolizumab pegol); Simponi (golimumab); Ilaris (canakinumab); Stelara (ustekinumab); Arzerra (ofatumumab); Prolia (denosumab); Numax (motavizumab); ABThrax (raxicumab); Benlysta (belimumab); Yervoy (ipilimumab); Adcetris (brentuximab); Perjeta (pertuzumab); Kadcyla (trastuzumab-emtansine conjugate); and Gazyva (atazolizumab). Antibody-drug conjugates are also included.

[0591] The second agent can be a non-drug therapeutic agent. For example, the second therapeutic agent is radiation therapy, cryotherapy, hyperthermia and / or surgical resection of tumor tissue.

[0592] The second agent can be a checkpoint inhibitor. In one embodiment, the checkpoint inhibitor is an inhibitory antibody (e.g., a monospecific antibody such as a monoclonal antibody). The antibody can be, for example, humanized or fully human. In certain embodiments, the checkpoint inhibitor is a fusion protein, for example, an Fc-receptor fusion protein. In certain embodiments, the checkpoint inhibitor is an agent that interacts with a checkpoint protein, such as an antibody. In certain embodiments, the checkpoint inhibitor is an agent that interacts with a ligand of a checkpoint protein, such as an antibody. In certain embodiments, the checkpoint inhibitor 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 / Yervoy or tremelimumab). In certain embodiments, the checkpoint inhibitor is an inhibitor of PD-1 (e.g., an inhibitory antibody or a small molecule inhibitor) (e.g., nivolumab / Opdivo®; pembrolizumab / Keytruda®; pidilizumab / CT-011). In certain embodiments, the checkpoint inhibitor is an inhibitor (e.g., an inhibitory antibody or small molecule inhibitor) of PDL1 (e.g., MPDL3280A / RG7446; MEDI4736; MSB0010718C; BMS 936559). In certain embodiments, the checkpoint inhibitor is an inhibitor (e.g., an inhibitory antibody or Fc fusion or small molecule inhibitor) of PDL2 (e.g., a PDL2 / Ig fusion protein such as AMP 224). In certain embodiments, the checkpoint inhibitor is an inhibitor (e.g., an inhibitory antibody or small molecule inhibitor) of B7-H3 (e.g., MGA271), B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK 1, CHK2, A2aR, B-7 family ligand, or a combination thereof.

[0593] In any combination embodiment described herein, the first and second therapeutic agents are administered simultaneously or sequentially in either order. The first therapeutic agent can be administered immediately, up to 1 hour, up to 2 hours, up to 3 hours, up to 4 hours, up to 5 hours, up to 6 hours, up to 7 hours, up to 8 hours, up to 9 hours, up to 10 hours, up to 11 hours, up to 12 hours, up to 13 hours, 14 hours, up to 16 hours, up to 17 hours, up to 18 hours, up to 19 hours up to 20 hours, up to 21 hours, up to 22 hours, up to 23 hours up to 24 hours, or up to 1-7, 1-14, 1-21, or 1-30 days before or after the second therapeutic agent.

[0594] Pharmaceutical composition

[0595] The compounds of the present invention are preferably formulated into pharmaceutical compositions and administered to mammals, preferably humans, in a biocompatible form suitable for in vivo administration. Thus, in one aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention admixed with a suitable diluent, carrier or excipient.

[0596] The compound of the present invention can be used in the form of free alkali, in the form of salt, solvate and as prodrug.All forms are within the scope of the present invention.As will be appreciated by those skilled in the art, according to the method of the present invention, described compound or its salt, solvate or prodrug can be administered to the patient in various forms, depending on the selected route of administration.The compound of the present invention can be used, for example, by oral, parenteral, buccal, sublingual, nasal, rectal, patch, pump or transdermal administration, and pharmaceutical composition is formulated accordingly. Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, epithelial, nasal, intrapulmonary, intrathecal, rectal and topical administration modes. Parenteral administration can be carried out by continuous infusion in selected time period.

[0597] The compounds of the present invention can be administered orally, for example, with an inert diluent or with an assimilable edible carrier, or they can be encapsulated in a hard or soft shell gelatin capsule, or they can be compressed into tablets, or they can be directly admixed with food in the diet. For oral therapy, the compounds of the present invention can be admixed with excipients and used in the form of ingestible tablets, buccal tablets, lozenges, capsules, elixirs, suspensions, syrups, and wafers. The compounds of the present invention can also be administered parenterally. Solutions of the compounds of the present invention can be prepared in water suitably mixed with a surfactant (such as hydroxypropylcellulose). Dispersions can also be prepared in glycerol, liquid polyethylene glycol, DMSO, and mixtures thereof with or without alcohol, and in oils. Under common storage and use conditions, these preparations can contain preservatives to prevent the growth of microorganisms. Conventional procedures and ingredients for selecting and preparing suitable formulations are described in, for example, Remington's Pharmaceutical Sciences (2003, 20th edition) and The United States Pharmacopeia: The National Formulary (USP 24 NF19) disclosed in 1999. Pharmaceutical forms suitable for injection include sterile aqueous solutions or dispersions, and sterile powders for the immediate preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must be a fluid that can be easily administered via a syringe. Compositions for nasal administration can be conveniently formulated into aerosols, drops, gels, and powders. Aerosol formulations typically include solutions or fine suspensions of active substances in physiologically acceptable aqueous or non-aqueous solvents, and are typically present in a sealed container in a sterile form in a single dose or multiple dose amount, which container can be in the form of a cartridge or refilled for use with a nebulizer. Alternatively, the sealed container can be a unit dispensing device, such as a single-dose nasal inhaler or an aerosol dispenser equipped with a metering valve that is intended to be discarded after use. Where the dosage form comprises 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 fluorochlorocarbon. Aerosol dosage forms 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, gum tragacanth, gelatin, and glycerin. Compositions for rectal administration are conveniently in the form of suppositories containing conventional suppository bases, such as cocoa butter. The compounds described herein can be administered intratumorally, for example, as intratumoral injections. Intratumoral injections are injections directly into the tumor vasculature and are particularly contemplated for use in discrete, solid, accessible tumors. Local, regional, or systemic administration may also be appropriate.The compounds described herein can be advantageously contacted by administering a single injection or multiple injections to the tumor, for example spaced about 1 cm apart. In the case of surgical intervention, the present invention can be used prior to surgery, such as to allow resection of an inoperable tumor. Continuous administration can also be used where appropriate, for example, by implanting a catheter into the tumor or tumor vasculature.

[0598] As indicated herein, the compounds of the invention can be administered to animals, such as humans, alone or in combination with a pharmaceutically acceptable carrier, in proportions determined by the solubility and chemical properties of the compound, the chosen route of administration, and standard pharmaceutical practice.

[0599] dose

[0600] The dosage of the compounds of the present invention and / or compositions comprising the compounds of the present invention may vary depending on many factors, such as the pharmacodynamic properties of the compound; the mode of administration; the age, health and weight of the recipient; the nature and extent of 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. One skilled in the art can determine the appropriate dosage based on the above factors. The compounds of the present invention may be initially administered at a suitable dosage, which may be adjusted as needed based on the clinical response. In general, satisfactory results may be obtained when the compounds of the present invention are administered to humans at a daily dose of, for example, 0.05 mg to 3000 mg (measured in solid form). Dosage ranges include, for example, 10-1000 mg (e.g., 50-800 mg). In certain 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.

[0601] Alternatively, the patient's weight can be used to calculate the dosage. For example, the dosage of the compound or its pharmaceutical composition administered to the patient can be in the range of 0.1-100 mg / kg (e.g., 0.25-25 mg / kg). In exemplary, non-limiting embodiments, the dosage can be in the range of 0.5-5.0 mg / kg (e.g., 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, or 5.0 mg / kg) or 5.0-20 mg / kg (e.g., 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg / kg). Example

[0602] The following abbreviations are used in the following examples.

[0603] Ac acetyl

[0604] ACN or MeCN Acetonitrile

[0605] AcOH acetic acid

[0606] Ac2O acetic anhydride

[0607] aq. water-based

[0608] Boc tert-Butoxycarbonyl

[0609] Bu or n-Bu butyl

[0610] CDI 1,1′-Carbonyldiimidazole

[0611] DCE or 1,2-DCE 1,2-dichloroethane

[0612] DCM dichloromethane

[0613] DIAD Diisopropyl azodicarboxylate

[0614] DIPEA or DIEA NN-diisopropylethylamine

[0615] DMAP 4-(dimethylamino)pyridine

[0616] DME 1,2-dimethoxyethane

[0617] DMF NN-dimethylformamide

[0618] DMSO dimethyl sulfoxide

[0619] EA or EtOAc Ethyl acetate

[0620] EDCI N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride

[0621] equiv equivalent

[0622] Et3N or TEA triethylamine

[0623] EtOH

[0624] FA Formic acid

[0625] h or hr hours

[0626] HATU 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate

[0627] HOAt 1-Hydroxy-7-azabenzotriazole

[0628] HOBt or HOBT 1-hydroxybenzotriazole hydrate

[0629] iPr Isopropyl

[0630] MeOH methanol

[0631] Me4t-BuXphos Di-tert-butyl-[2,3,4,5-tetramethyl-6-(2,4,6-triisopropylphenyl)phenyl]phosphane

[0632] min

[0633] MTBE tert-butyl methyl ether

[0634] n-BuLi n-butyllithium

[0635] NMP 1-Methyl-2-pyrrolidone

[0636] OAc acetate

[0637] Pd / C Palladium on Carbon

[0638] PDC Pyridinium Dichromate

[0639] PdCl2(dtbpf) or Pd(dtbpf)Cl2 dichloro[1,1'-bis(di-tert-butylphosphino)ferrocene]palladium(II)

[0640] PdCl2(dppf) or Pd(dppf)Cl2 [1,1′-bis(diphenylphosphino)ferrocene]palladium(II) dichloride

[0641] Pd2(dba)3 tris(dibenzylideneacetone)dipalladium(0)

[0642] Pd(PPh3)4tetrakis(triphenylphosphine)palladium(0

[0643] Pd(PPh3)2Cl2 Dichlorobis(triphenylphosphine)palladium(II)

[0644] PE petroleum ether

[0645] PPh3 triphenylphosphine

[0646] Pr n-propyl

[0647] Py pyridine

[0648] rac racemic

[0649] Rf retention factor

[0650] rt or rt room temperature

[0651] sat. saturated

[0652] SFC Supercritical Fluid Chromatography

[0653] t-Bu tert-butyl

[0654] tBuXphos-Pd-G3 or tBuXphos PdG3 or t-BuXphos-Pd (gen 3) [2-(2-aminophenyl)phenyl]-methanesulfonyloxypalladium; di-tert-butyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphane

[0655] TFA trifluoroacetic acid

[0656] Tf2O trifluoromethanesulfonic anhydride

[0657] THF Tetrahydrofuran

[0658] TLC thin layer chromatography

[0659] Xantphos-Pd-G3 [2-(2-aminophenyl)phenyl]-methylsulfonyloxy-palladium; (5-diphenylphosphanyl-9,9-dimethyl-xanthen-4-yl)-diphenyl-phosphane.

[0660] Example 1. Preparation of intermediates

[0661] Preparation of (2S,4R)-1-[(2S)-2-(10-aminodecanoylamino)-3,3-dimethylbutyryl]-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (I-1)

[0662]

[0663] Step 1: N-(9-[[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazol-5-yl)benzene 1-yl]methyl]carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamoyl]nonyl)amino Preparation of tert-butyl ester

[0664]

[0665] To a stirred mixture 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 (1.00 g, 0.002 mmol, 1.00 equiv) and 10-[(tert-butoxycarbonyl)amino]decanoic acid (0.73 g, 0.003 mmol, 1.10 equiv) in DCM (20.00 mL) was added DIEA (0.90 g, 0.007 mmol, 3.00 equiv). The mixture was stirred at room temperature for 5 min, then HATU (1.32 g, 0.003 mmol, 1.50 equiv) was added. After stirring at room temperature for 2 h, water (100 mL) was added to the mixture and the mixture was extracted with DCM (100 mL x 4). The organic fractions were combined and dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated to produce the crude product, which was purified by chromatography on silica gel, eluting with DCM / MeOH (from a 100:0 DCM:MeOH ratio to a 100:7 DCM:MeOH ratio) to produce tert-butyl N-(9-[[(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]nonyl)carbamate (1.67 g, 92.46%). LCMS (ESI) m / z [M+H] + = 700.

[0666] Step 2: (2S,4R)-1-[(2S)-2-(10-aminodecanoylamino)-3,3-dimethylbutanoyl]-4-hydroxy- Preparation of N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (I-1)

[0667]

[0668] To a stirred solution of tert-butyl N-(9-[[(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]nonyl)carbamate (1.67 g, 2.386 mmol, 1 eq) in DCM (10 mL) was added TFA (2 mL, 26.926 mmol, 11.29 eq) and the mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under vacuum. A 5% KCO (MeOH / water = 5 / 2) solution was added and the pH was adjusted to 8-9. The mixture was stirred at room temperature for 2 h. The final mixture was concentrated under vacuum and the crude material was purified by reverse phase flash chromatography (C18 silica gel column) eluting with 10-35% acetonitrile in 0.1% aqueous NH4HCO3 over 20 min, detecting at UV 254 nm to provide (2S,4R)-1-[(2S)-2-(10-aminodecanoylamino)-3,3-dimethylbutanoyl]-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (I-1, 1.11 g, 73.68%). LCMS (ESI) m / z: [M+H] + = 600.40. The following intermediates in Table A1 were prepared by a route similar to that used to prepare intermediate I-1.

[0669]

[0670] Preparation of 5-(2-(4-aminopiperidin-1-yl)ethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione FA (I-6)

[0671]

[0672] Step 1: Preparation of 5-(2-bromoethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0673]

[0674] To a solution of 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindole-1,3-dione (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), PPh3 (1.97 g, 7.494 mmol, 1.5 equiv) and DIAD (1.52 g, 7.494 mmol, 1.5 equiv) at 0° C. The resulting mixture was stirred at room temperature for 2 h. The residue was purified by reverse phase flash chromatography (C18 silica gel column) eluting with 0-100% ACN in water over 45 min to provide 5-(2-bromoethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (1.52 g, 79.82%) as a black solid. LCMS (ESI) m / z: [M+H] + = 381.38.

[0675] Step 2: (1-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy) Preparation of tert-butyl (4-ethyl)piperidin-4-yl)carbamate

[0676]

[0677] To a solution of 5-(2-bromoethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (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 KCO (1.65 g, 11.963 mmol, 3.00 equiv). The resulting solution was stirred at 70°C for 2 h. The residue was purified by reverse phase flash chromatography (C18 silica gel), eluting with 0-100% ACN in water over 30 min to afford tert-butyl (1-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethyl)piperidin-4-yl)carbamate (1.402 g, 70.24%) as a colorless solid. LCMS (ESI) m / z: [M+H] + = 501.

[0678] Step 3: 5-(2-(4-aminopiperidin-1-yl)ethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline- Preparation of 1,3-diketone FA (I-6)

[0679]

[0680] To a solution of tert-butyl (1-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethyl)piperidin-4-yl)carbamate (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 and then concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography (C18 silica gel column), eluting with 0-100% ACN in 0.1% formic acid / water over 45 min to yield 5-(2-(4-aminopiperidin-1-yl)ethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione formate (I-6, 840 mg, 62.52%) as a white solid. LCMS (ESI) m / z: [M+H] + = 401.17.

[0681] The following intermediates in Table A2 were prepared by routes similar to those used to prepare intermediate 1-6.

[0682]

[0683] Preparation of 4-(azetidin-3-ylmethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione TFA (I-7)

[0684]

[0685] Step 1: Preparation of tert-butyl 3-((tosyloxy)methyl)azetidine-1-carboxylate

[0686]

[0687] To a stirred solution of tert-butyl 3-(hydroxymethyl)azetidine-1-carboxylate (1.87 g, 9.987 mmol, 1.00 equiv) in DCM (50.00 mL, 786.502 mmol, 78.75 equiv) was added 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) at 0° C. The resulting mixture was stirred at 0° C. for 2 h, then allowed to warm to room temperature and stirred for an additional 5 h. The residue was purified by silica gel column chromatography eluting with petroleum ether / THF (1:1) to provide tert-butyl 3-((tosyloxy)methyl)azetidine-1-carboxylate (2.65 g, 77.72%) as a colorless oil. LCMS (ESI) m / z: [M+H] + = 342.

[0688] Step 2: 3-([[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]methyl) Preparation of tert-butyl azetidine-1-carboxylate

[0689]

[0690] To a solution of tert-butyl 3-((tosyloxy)methyl)azetidine-1-carboxylate (2.30 g, 8.387 mmol, 1.00 equiv) in DMF (15.00 mL) was added 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione (2.86 g, 8.387 mmol, 1 equiv) and Na2CO3 (1.33 g, 12.581 mmol, 1.5 equiv). The resulting mixture was stirred at 80 ° C for 5 h under an atmosphere of dry nitrogen. The reaction was quenched with water at room temperature. The resulting mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (3 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to give tert-butyl 3-([[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]methyl)azetidine-1-carboxylate (3.37 g, 90.61%) as a pale yellow solid. LCMS (ESI) m / z: [M+H] + = 444.

[0691] Step 3: 4-(azetidin-3-ylmethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3- Preparation of diketone TFA (I-7)

[0692]

[0693] To a solution of tert-butyl 3-([[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]methyl)azetidine-1-carboxylate (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. The residue was purified by reverse phase flash chromatography (C18 silica gel column) eluting with 0-100% ACN in water over 45 min to provide 4-(azetidin-3-ylmethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione TFA (I-7, 1.712 g, 87.72%) as a light yellow solid. LCMS (ESI) m / z: [M+H] + = 344.12.

[0694] The following intermediates in Table A3 were prepared by routes similar to those used to prepare intermediate 1-7.

[0695]

[0696] Preparation of 5-(azetidin-3-ylmethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione TFA (I-8)

[0697]

[0698] Step 1: 3-([[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]methyl) Preparation of tert-butyl azetidine-1-carboxylate

[0699]

[0700] To a solution of tert-butyl 3-((tosyloxy)methyl)azetidine-1-carboxylate (2.50 g, 9.116 mmol, 1.00 equiv) in DMF (15.00 mL) was added 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione (3.11 g, 9.116 mmol, 1 equiv) and Na2CO3 (1.45 g, 13.675 mmol, 1.5 equiv). The resulting solution was stirred at 80 ° C for 5 h under an atmosphere of dry nitrogen. The reaction was cooled to room temperature and quenched with water, then extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (3 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. This resulted in tert-butyl 3-([[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]methyl)azetidine-1-carboxylate (2.15 g, 53.18%) as a light yellow solid. LCMS (ESI) m / z: [M+H] + = 444.

[0701] Step 2: 5-(azetidin-3-ylmethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-diol Preparation of Ketone TFA (I-8)

[0702]

[0703] To a solution of tert-butyl 3-([[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]methyl)azetidine-1-carboxylate (1.41 g, 3.180 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 then concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography (C18 silica gel column), eluting with 0-100% ACN in water over 45 min to provide 5-(azetidin-3-ylmethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione TFA (I-8, 906.2 mg, 81.38%) as a white solid. LCMS (ESI) m / z: [M+H] + = 344.12.

[0704] Preparation of 4-((3-((4-aminobutyl)sulfonyl)propyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione FA (I-9)

[0705]

[0706] Step 1: Preparation of tert-butyl N-(4-hydroxybutyl)carbamate

[0707]

[0708] A solution of di-tert-butyl decarbonate (52.89 g, 242.321 mmol, 1.5 equiv) and 4-aminobutan-1-ol (14.40 g, 161.547 mmol, 1.00 equiv) in THF (160.00 mL) was stirred at room temperature for 1 h. The solution was concentrated to dryness, and the oily residue was purified by flash column chromatography (eluting with 40-60% EtOAc-hexanes) to provide tert-butyl N-(4-hydroxybutyl)carbamate as a colorless oil that solidified to a white solid upon standing. LCMS (ESI) m / z: [M+H] + = 190.

[0709] Step 2: Preparation of 4-((tert-butoxycarbonyl)amino)butyl 4-methylbenzenesulfonate

[0710]

[0711] To a stirred solution of tert-butyl N-(4-hydroxybutyl)carbamate (30.58 g, 161.581 mmol, 1.00 equiv) in DCM (400.00 mL) at 0°C was added DMAP (2.96 g, 24.237 mmol, 0.15 equiv), TEA (40.88 g, 403.952 mmol, 2.5 equiv), and p-toluenesulfonyl chloride (46.21 g, 242.371 mmol, 1.5 equiv). The resulting mixture was stirred at 0°C for 2 h, then at room temperature for 5 h. The solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography, eluting with petroleum ether / THF (1:1) to provide 4-(tert-butoxycarbonyl)amino)butyl 4-methylbenzenesulfonate (45.6 g, 82.17%) as a pale yellow oil. LCMS (ESI) m / z: [M+H] + = 344.

[0712] Step 3: Preparation of S-(4-((tert-butoxycarbonyl)amino)butyl)thioacetate

[0713]

[0714] To a stirred solution of 4-((tert-butoxycarbonyl)amino)butyl 4-methylbenzenesulfonate (45.60 g, 132.774 mmol, 1.00 equiv) in ACN (300.00 mL) was added thioacetic acid (15.16 g, 199.161 mmol, 1.5 equiv) and KCO (55.05 g, 398.323 mmol, 3 equiv). The resulting mixture was stirred at room temperature for 12 h. The mixture was concentrated under reduced pressure and the residue was purified by silica gel column chromatography eluting with petroleum ether / THF (1:1) to afford S-(4-((tert-butoxycarbonyl)amino)butyl)thioacetate (28.7 g, 87.39%) as a light yellow oil. LCMS (ESI) m / z: [M+H] + = 248.

[0715] Step 4: Preparation of benzyl N-[3-([4-[(tert-butoxycarbonyl)amino]butyl]sulfanyl)propyl]carbamate Preparation

[0716]

[0717] To a solution of S-(4-((tert-butoxycarbonyl)amino)butyl)thioacetate (3.60 g, 14.554 mmol, 1.00 equivalent) in MeOH (90.00 mL, 2222.902 mmol, 152.73 equivalents) was added benzyl (3-bromopropyl)carbamate (4.36 g, 16.010 mmol, 1.1 equivalents) and NaOMe (3.15 g, 58.217 mmol, 4 equivalents). The resulting solution was stirred at room temperature for 3 h. The reaction was quenched with water at room temperature. The resulting mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (3 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography (C18 silica gel column) eluting with 0-100% ACN in aqueous NH4HCO3 to provide benzyl N-[3-([4-[(tert-butoxycarbonyl)amino]butyl]sulfanyl)propyl]carbamate (4.122 g, 71.42%) as a light yellow oil. LCMS (ESI) m / z: [M+H] + = 397.

[0718] Step 5: Preparation of benzyl N-(3-[4-[(tert-butoxycarbonyl)amino]butanesulfonyl]propyl)carbamate

[0719]

[0720] To a solution of benzyl N-[3-([4-[(tert-butoxycarbonyl)amino]butyl]sulfanyl)propyl]carbamate (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 and then concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography (C18 silica gel column), eluting with 0-100% ACN in aqueous NH4HCO3 over 30 min to provide benzyl N-(3-[4-[(tert-butoxycarbonyl)amino]butanesulfonyl]propyl)carbamate (2 g, 44.81%) as a white solid. LCMS (ESI) m / z: [M+H] + = 429.

[0721] Step 6: Preparation of tert-butyl (4-((3-aminopropyl)sulfonyl)butyl)carbamate

[0722]

[0723] To a solution of benzyl N-(3-[4-[(tert-butoxycarbonyl)amino]butanesulfonyl]propyl)carbamate (1.95 g, 4.550 mmol, 1.00 equiv) in EtOH (30.00 mL) was added ammonium formate (573.85 mg, 9.101 mmol, 2 equiv) and 5% Pd(OH)2 / C (977.68 mg, 6.962 mmol, 1.53 equiv). The resulting solution was stirred at 60 °C under 1 atmosphere of hydrogen for 12 h. The resulting mixture was filtered, the filter cake was washed with MeOH (3 x 30 mL), and the filtrate was concentrated under reduced pressure to afford tert-butyl (4-((3-aminopropyl)sulfonyl)butyl)carbamate (1.12 g, crude) as a black solid. LCMS (ESI) m / z: [M+H] + = 295.

[0724] Step 7: (4-((3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino) Preparation of tert-butyl (propyl)sulfonyl)butyl)carbamate

[0725]

[0726] To a solution of tert-butyl (4-((3-aminopropyl)sulfonyl)butyl)carbamate (1.12 g, 3.804 mmol, 1.00 equiv) in NMP (30.00 mL) was added 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindole-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 for 3 h under an atmosphere of dry nitrogen. The resulting mixture was diluted with water (30 mL) and extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (3 x 30 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography (C18 silica gel column), eluting with 0-100% ACN in 0.1% formic acid / water over 45 min to afford tert-butyl (4-((3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)propyl)sulfonyl)butyl)carbamate as a yellow solid (970 mg, 46.31%). LCMS (ESI) m / z: [M+H] + = 551.

[0727] Step 8: 4-((3-((4-aminobutyl)sulfonyl)propyl)amino)-2-(2,6-dioxopiperidin-3-yl)isothiazolin Preparation of indoline-1,3-dione FA (I-9)

[0728]

[0729] To a solution of tert-butyl (4-((3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)propyl)sulfonyl)butyl)carbamate (970.00 mg, 1.762 mmol, 1.00 equiv) was added 4N dry HCl in dioxane (5.00 mL, 164.559 mmol, 93.41 equiv). The resulting solution was stirred at room temperature for 3 h and the solvent was removed under reduced pressure. The residue was purified by reverse phase flash chromatography (C18 silica gel column) eluting with 0-100% ACN in 0.1% formic acid / water over 45 min to afford 4-((3-((4-aminobutyl)sulfonyl)propyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione FA (I-9, 712 mg, 89.17%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 451.16.

[0730] Preparation of 4-[[2-(2-aminoethanesulfonyl)ethyl]amino]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione TFA (I-10)

[0731]

[0732] Step 1: tert-Butyl N-(2-[[2-(1,3-dioxoisoindol-2-yl)ethyl]sulfanyl]ethyl)carbamate Preparation of esters

[0733]

[0734] To a stirred mixture of tert-butyl N-(2-sulfanylethyl)carbamate (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) at 70° C. under an atmosphere of dry nitrogen was added KCO (11.70 g, 0.085 mmol, 3.00 equiv). After 5 h, the resulting mixture was extracted with EtOAc (3 x 500 mL). The combined organic layers were washed with brine (3 x 300 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with petroleum ether / EtOAc (5:1 to 1:1) to provide tert-butyl N-(2-[[2-(1,3-dioxoisoindol-2-yl)ethyl]sulfanyl]ethyl)carbamate (8.20 g, 82.96%) as a white solid. LCMS (ESI) m / z: [M+H] + =351.

[0735] Step 2: Preparation of tert-butyl N-[2-[2-(1,3-dioxoisoindol-2-yl)ethanesulfonyl]ethyl]carbamate preparation

[0736]

[0737] To a stirred mixture of tert-butyl N-(2-[[2-(1,3-dioxoisoindol-2-yl)ethyl]sulfanyl]ethyl)carbamate (8.20 g, 23.400 mmol, 1.00 equiv) in DCM (100 mL) was added m-CPBA (12.11 g, 70.199 mmol, 3.00 equiv) at room temperature under an atmosphere of dry nitrogen. The reaction was then quenched with saturated aqueous Na2S2O3 (100 mL) at room temperature. Saturated aqueous NaHCO3 (100 mL) was added to the resulting mixture and the mixture was extracted with EtOAc (3 x 400 mL). The organic phase was separated 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 petroleum ether / EtOAc (5:1 to 1:1) to provide tert-butyl N-[2-[2-(1,3-dioxoisoindol-2-yl)ethanesulfonyl]ethyl]carbamate (8.40 g, 87.30%) as a white solid. LCMS (ESI) m / z: [M+H] + = 383.

[0738] Step 3: Preparation of tert-butyl N-[2-(2-aminoethanesulfonyl)ethyl]carbamate

[0739]

[0740] To a stirred mixture of tert-butyl N-[2-[2-(1,3-dioxoisoindol-2-yl)ethanesulfonyl]ethyl]carbamate (3.40 g, 8.891 mmol, 1.00 equiv) in EtOH (100 mL) at 80 °C under an atmosphere of dry nitrogen was added hydrazine hydrate (0.89 g, 17.781 mmol, 2.00 equiv). The resulting mixture was stirred at 80 °C under an atmosphere of dry nitrogen for 1 h. The resulting mixture was filtered, the filter cake was washed with EtOH (100 mL), and the filtrate was concentrated under reduced pressure to afford tert-butyl N-[2-(2-aminoethanesulfonyl)ethyl]carbamate (1.88 g, 77.94%) as a white solid. LCMS (ESI) m / z: [M+H] + = 253.

[0741] Step 4: (2-((2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino) Preparation of tert-butyl (ethyl)sulfonyl)ethyl)carbamate

[0742]

[0743] To a stirred mixture of tert-butyl N-[2-(2-aminoethanesulfonyl)ethyl]carbamate (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 equivalent) in NMP (25.00 mL) was added DIEA (2.89 g, 22.352 mmol, 3.00 equivalent) dropwise at 90 ° C under an atmosphere of dry nitrogen, and the solution was stirred for 12 hours. The resulting mixture was cooled and extracted with EtOAc (3 x 300 mL). The combined organic layers were washed with saturated brine (3 x 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 petroleum ether / EtOAc (1:1) to afford tert-butyl (2-((2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethyl)sulfonyl)ethyl)carbamate (1.58 g, 40.03%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 509.

[0744] Step 5: 4-[[2-(2-aminoethanesulfonyl)ethyl]amino]-2-(2,6-dioxopiperidin-3-yl)isoindole Preparation of Indole-1,3-dione TFA (I-10)

[0745]

[0746] To a stirred mixture of tert-butyl (2-((2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethyl)sulfonyl)ethyl)carbamate (1.54 g, 3.028 mmol, 1.00 equiv) in DCM (20 mL) at 25° C. under an atmosphere of dry nitrogen was added TFA (5.0 mL) dropwise. After 1 hour, the resulting mixture was concentrated under vacuum to give intermediate I-10 (1.68 g, 122.25%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 409.11.

[0747] Preparation of 3-[5-([2-[2-(2-aminoethoxy)ethoxy]ethyl]amino)-2-methyl-4-oxoquinazolin-3-yl]piperidine-2,6-dione TFA (I-11)

[0748]

[0749] Step 1: (2-(2-(2-((3-(2,6-dioxopiperidin-3-yl)-2-methyl-4-oxo-3,4-dihydroquinazole Preparation of tert-butyl (5-(1-(1-(1-(2-((1-((2-( ...

[0750]

[0751] To a stirred solution of 4-hydroxy-2,2-dimethyl-3,8,11-trioxa-5-azatridecan-13-al (1.25 g, 4.995 mmol, 1 eq) and 3-(5-amino-2-methyl-4-oxoquinazolin-3-yl)piperidine-2,6-dione (1.43 g, 4.995 mmol, 1.00 eq) in MeOH (30.00 mL) was added NaBH3CN (0.63 g, 9.990 mmol, 2 eq) at room temperature and the resulting mixture was stirred for 1 h. The reaction was quenched with saturated aqueous NH4Cl at 0°C, the solvent was evaporated, and the resulting residue was purified by silica gel column chromatography eluting with petroleum ether / EtOAc (9:1) to afford tert-butyl (2-(2-(2-((3-(2,6-dioxopiperidin-3-yl)-2-methyl-4-oxo-3,4-dihydroquinazolin-5-yl)amino)ethoxy)ethoxy)ethyl)carbamate (1.29 g, 49.70%) as a yellow solid. LCMS (ESI) m / z [M+H] + =518.

[0752] Step 2: 3-[5-([2-[2-(2-aminoethoxy)ethoxy]ethyl]amino)-2-methyl-4-oxoquinazole Preparation of [3-[[(1- ...(-[(-(-(-(-(-(-(-(-(-(piperidine-3-ylpiperidine-2,6-dione(TFA ((piperidine-2,6-dione((1-11

[0753]

[0754] A solution of tert-butyl (2-(2-(2-((3-(2,6-dioxopiperidin-3-yl)-2-methyl-4-oxo-3,4-dihydroquinazolin-5-yl)amino)ethoxy)ethoxy)ethyl)carbamate (1.29 g, 2.483 mmol, 1.00 equiv) and TFA (8.49 g, 74.481 mmol, 30 equiv) in DCM (6.00 mL) was stirred at room temperature for 1 h. The reaction mixture was concentrated, and the resulting residue was purified by reverse phase flash chromatography using the following conditions (column: C18 silica gel; mobile phase: ACN in water, 10% to 50% gradient over 10 min; detector, UV 254 nm) to provide 3-[5-([2-[2-(2-aminoethoxy)ethoxy]ethyl]amino)-2-methyl-4-oxoquinazolin-3-yl]piperidine-2,6-dione TFA (I-11, 1.56 g, 95.13%) as a light brown solid. LCMS (ESI) m / z: [M+H] + = 418.20.

[0755] Preparation of 3-[4-([2-[2-(2-aminoethoxy)ethoxy]ethyl]amino)-1-oxo-3H-isoindol-2-yl]piperidine-2,6-dione TFA (I-12)

[0756]

[0757] Step 1: (2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)ethyl Preparation of tert-butyl (1,2-dioxy)ethoxy)ethyl)carbamate

[0758]

[0759] To a stirred solution of tert-butyl (2-(2-(2-oxoethoxy)ethoxy)ethyl)carbamate (1.33 g, 0.005 mmol, 1.00 equiv) and lenalidomide (1.38 g, 5.323 mmol, 1.00 equiv) in MeOH (20.00 mL) was added NaBH3CN (0.67 g, 0.011 mmol, 2.00 equiv) at room temperature and the resulting mixture was stirred for 1 h. The reaction was then quenched with saturated aqueous NH4Cl at 0°C, the solvent was evaporated, and the resulting residue was purified by silica gel column chromatography eluting with petroleum ether / EtOAc (9:1) to provide tert-butyl (2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethyl)carbamate (0.92 g, 34.12%) as a yellow solid. LCMS (ESI) m / z [M+H] + = 491.

[0760] Step 2: 3-[4-([2-[2-(2-aminoethoxy)ethoxy]ethyl]amino)-1-oxo-3H-isoindole- Preparation of 2-amino-1,6-piperidine-2,6-dione TFA (I-12)

[0761]

[0762] A solution of tert-butyl (2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethyl)carbamate (1.20 g, 2.436 mmol, 1.00 equiv) and TFA (5.56 g, 48.724 mmol, 20.00 equiv) in DCM (5.00 mL) was stirred at room temperature for 1 h. The reaction mixture was concentrated and the resulting residue was purified by reverse phase flash chromatography (C18 silica gel column), eluting with 10-50% ACN in water over 10 min and detecting at UV 254 nm to afford 3-[4-([2-[2-(2-aminoethoxy)ethoxy]ethyl]amino)-1-oxo-3H-isoindol-2-yl]piperidine-2,6-dione TFA (I-12, 979 mg, 93.87%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 391.19.

[0763] Preparation of 3-(5-((6-aminohexyl)amino)-2-methyl-4-oxoquinazolin-3(4H)-yl)piperidine-2,6-dione trifluoroacetate (I-13)

[0764]

[0765] Step 1: Preparation of tert-butyl N-(6-oxohexyl)carbamate

[0766]

[0767] To a stirred solution of tert-butyl (6-hydroxyhexyl)carbamate (800.00 mg, 3.681 mmol, 1.00 equiv) in DCM (10.00 mL) was added PCC (1190.31 mg, 5.522 mmol, 1.50 equiv) and alumina (75.07 mg, 0.736 mmol, 0.20 equiv). The resulting mixture was stirred at room temperature overnight. The solution was concentrated and the residue was purified by silica gel column chromatography, eluting with DCM / MeOH (10:1) to provide tert-butyl N-(6-oxohexyl)carbamate (500 mg, 63.09%) as a yellow oil.

[0768] Step 2: (6-((3-(2,6-dioxopiperidin-3-yl)-2-methyl-4-oxo-3,4-dihydroquinazoline-5-yl)- Preparation of tert-butyl)amino)hexyl)carbamate

[0769]

[0770] To a stirred solution of tert-butyl N-(6-oxohexyl)carbamate (380.00 mg, 1.765 mmol, 1.00 equiv) and 3-(5-amino-2-methyl-4-oxoquinazolin-3(4H)-yl)piperidine-2,6-dione (252.66 mg, 0.883 mmol, 0.50 equiv) in MeOH (5.00 mL) was added NaBHCN (221.84 mg, 3.530 mmol, 2.00 equiv). The resulting mixture was stirred at room temperature for 2 h. The solution was concentrated and purified by preparative TLC (eluting with 20:1 DCM / MeOH) to provide tert-butyl (6-((3-(2,6-dioxopiperidin-3-yl)-2-methyl-4-oxo-3,4-dihydroquinazolin-5-yl)amino)hexyl)carbamate (200 mg, 23.34%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 486.

[0771] Step 3: 3-(5-((6-aminohexyl)amino)-2-methyl-4-oxoquinazolin-3(4H)-yl)piperidin-2,6- Preparation of diketone TFA (I-13)

[0772]

[0773] To a stirred solution of tert-butyl (6-((3-(2,6-dioxopiperidin-3-yl)-2-methyl-4-oxo-3,4-dihydroquinazolin-5-yl)amino)hexyl)carbamate (100 mg) in DCM (1.00 mL) was added TFA (0.50 mL). The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated under reduced pressure to afford 3-(5-((6-aminohexyl)amino)-2-methyl-4-oxoquinazolin-3(4H)-yl)piperidine-2,6-dione trifluoroacetate (80 mg, 83.98%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 386.21.

[0774] Preparation of 3-(5-((8-aminooctyl)amino)-2-methyl-4-oxoquinazolin-3(4H)-yl)piperidine-2,6-dione TFA (I-14)

[0775]

[0776] Step 1: (8-((3-(2,6-dioxopiperidin-3-yl)-2-methyl-4-oxo-3,4-dihydroquinazoline-5-yl)- Preparation of tert-butyl)amino)octyl)carbamate

[0777]

[0778] To a stirred mixture of 3-(5-amino-2-methyl-4-oxoquinazoline-3(4H)-yl)piperidine-2,6-dione (500.00 mg, 1.746 mmol, 1.00 equiv) and tert-butyl (8-oxooctyl)carbamate (850.00 mg, 3.493 mmol, 2.00 equiv) in MeOH was added AcOH (350.00 mg, 5.828 mmol, 3.34 equiv) at room temperature. The resulting mixture was stirred at room temperature for 2 h and then NaBHCN (9.22 mg, 0.147 mmol, 2.00 equiv) was added at room temperature. The resulting mixture was stirred at room temperature for another 1 h. The residue was purified by reverse phase flash chromatography (C18 silica gel column), eluting with 0-100% ACN in water over 30 min to afford tert-butyl (8-((3-(2,6-dioxopiperidin-3-yl)-2-methyl-4-oxo-3,4-dihydroquinazolin-5-yl)amino)octyl)carbamate (420 mg, 46.82%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 514.

[0779] Step 2: 3-(5-((8-aminooctyl)amino)-2-methyl-4-oxoquinazolin-3(4H)-yl)piperidin-2,6- Preparation of diketone TFA (I-14)

[0780]

[0781] To a solution of tert-butyl (8-((3-(2,6-dioxopiperidin-3-yl)-2-methyl-4-oxo-3,4-dihydroquinazolin-5-yl)amino)octyl)carbamate (1.54 g, 2.998 mmol, 1.00 equiv) in DCM (10.00 mL) was added TFA (10.00 mL, 134.630 mmol, 44.90 equiv). The resulting solution was stirred at room temperature for 6 h. The solution was concentrated and the residue was purified by reverse phase flash chromatography (C18 silica gel column) eluting with 0-100% ACN in water over 35 min to provide 3-(5-((8-aminooctyl)amino)-2-methyl-4-oxoquinazolin-3(4H)-yl)piperidine-2,6-dione TFA (I-14, 1.4 g, 112.92%) as a light yellow solid. LCMS (ESI) m / z: [M+H] + =414.24.

[0782] Preparation of 3-(4-((8-aminooctyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15)

[0783]

[0784] Step 1: Preparation of tert-butyl (8-oxooctyl)carbamate

[0785]

[0786] To a stirred mixture of tert-butyl (8-hydroxyoctyl)carbamate (1.00 g, 4.076 mmol, 1.00 equiv) and PCC (878.51 mg, 4.076 mmol, 1.00 equiv) in MeOH (15.0 mL) was added Al2O3 (2.49 g, 24.454 mmol, 6.00 equiv) portionwise at room temperature and the suspension was stirred under an atmosphere of dry nitrogen for 2 h. The precipitated solid was collected by filtration and washed with DCM (10 x 10 mL). The filtrate was concentrated under vacuum and the mixture was used in the next step without further purification. LCMS (ESI) m / z: [M+H] + = 243.34.

[0787] Step 2: (8-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)octyl)amino Preparation of tert-butyl formate

[0788]

[0789] To a stirred mixture of tert-butyl (8-oxooctyl)carbamate (403.00 mg, 1.656 mmol, 1.00 equiv) and 3-(4-amino-1-oxoisoindolin-2-yl)piperidine-2,6-dione (429.36 mg, 1.656 mmol, 1.00 equiv) in MeOH (15.0 mL) was added portionwise NaBHCN (312.21 mg, 4.968 mmol, 3.00 equiv) and AcOH (0.01 mL, 0.158 mmol, 0.10 equiv) at room temperature. The resulting mixture was stirred under an atmosphere of dry nitrogen for 12 h and then filtered. The filter cake was washed with MeOH (10 x 10 mL). The filtrate was concentrated under reduced pressure. The resulting mixture was purified by preparative TLC (DCM / MeOH 40:1) to afford tert-butyl (8-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)octyl)carbamate (153 mg, 18.61%) as a dark green oil. LCMS (ESI) m / z: [M+H] + = 486.61.

[0790] Step 3: 3-(4-((8-aminooctyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I- 15) Preparation

[0791]

[0792] To a stirred mixture of tert-butyl (8-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)octyl)carbamate (153.00 mg, 0.314 mmol, 1.00 equiv) in DCM (4.00 mL) was added TFA (1.00 mL) at room temperature, and the solution was stirred under an atmosphere of dry nitrogen for 2 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (DCM / MeOH 40:1) to afford 3-(4-((8-aminooctyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 110 mg, 88.71%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 386.5.

[0793] Preparation of 4-((8-aminooctyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione TFA (I-16)

[0794]

[0795] Step 1: (8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)octane Preparation of tert-butyl (2-(2-methyl-1,2-d)carbamate

[0796]

[0797] To a stirred solution of 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (1.00 g, 3.620 mmol, 1.00 equiv) and tert-butyl (8-aminooctyl)carbamate (973.19 mg, 3.982 mmol, 1.10 equiv) in NMP (16 mL) was added DIEA (935.79 mg, 7.241 mmol, 2.00 equiv) at room temperature. The resulting mixture was stirred at 90 ° C overnight. The reaction was cooled to room temperature and quenched with water (10 mL). The aqueous layer was extracted with EtOAc (3 x 50 mL) and the combined organic phases were dried over sodium sulfate, filtered, and the filtrate was concentrated. The residue was purified by silica gel column chromatography eluting with petroleum ether / EtOAc (1:1) to afford tert-butyl (8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)octyl)carbamate (1.19 g, 65.66%) as a light yellow solid. LCMS (ESI) m / z: [M+H] + = 501.

[0798] Step 2: 4-((8-aminooctyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione Preparation of TFA (I-16)

[0799]

[0800] To a stirred solution of tert-butyl (8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)octyl)carbamate (300 mg, 0.599 mmol, 1 eq) in DCM (10.00 mL) was added TFA (2.00 mL) dropwise at room temperature. The resulting mixture was stirred at room temperature for 4 h and then concentrated under reduced pressure. The residue was purified by CombiFlash to afford 4-((8-aminooctyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione TFA (I-16, 130 mg, 54.17%) as a yellow solid. LCMS (ESI) m / z: [M+H] + =401.15.

[0801] Preparation of 4-(2-([4,4'-bipiperidinyl]-1-yl)-2-oxoethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (I-17)

[0802]

[0803] Step 1: 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetic acid Preparation of tert-butyl ester

[0804]

[0805] To a stirred mixture of 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindolin-1,3-dione (10 g, 36.5 mmol, 1.0 eq) and KCO (7.56 g, 54.7 mmol, 1.5 eq) in ACN (300 mL) was added tert-butyl 2-bromoacetate (7.82 g, 40.1 mmol, 1.1 eq) at room temperature. The resulting mixture was stirred at room temperature for 2 h and then concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography to provide tert-butyl 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetate (5.32 g, 37.56%) as a white solid. LCMS (ESI) m / z [M+H] + =389.

[0806] Step 2: 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetic acid Preparation

[0807]

[0808] To a stirred mixture of tert-butyl 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetate (5.32 g, 13.69 mmol, 1.00 equiv) in DCM (20 mL) was added TFA (4.0 mL) dropwise at room temperature. The resulting mixture was stirred at room temperature for 2 h. The crude solution was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography to afford 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetic acid (4.66 g, quantitative) as a white solid. LCMS (ESI) m / z [M+H] + = 333.

[0809] Step 3: 1'-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy) Preparation of tert-butyl (4,4'-bipiperidinyl)-1-carboxylate

[0810]

[0811] To a stirred mixture of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetic acid (1.10 g, 3.311 mmol, 1.00 equiv) in DMF (10 mL) at room temperature under an atmosphere of dry nitrogen was added DIEA (8.54 g, 6.622 mmol, 2.00 equiv) and HATU (1.637 g, 4.30 mmol, 1.30 equiv). The resulting mixture was stirred for 30 min and then tert-butyl [4,4-bipiperidine]-1-carboxylate (0.89 g, 3.311 mmol, 1.0 equiv) was added at room temperature. The resulting mixture was stirred for an additional 1 h and then quenched with water (20 mL). The resulting mixture was extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography to provide tert-butyl 1'-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetyl)-[4,4'-bipiperidine]-1-carboxylate (1.36 g, 70.51%) as a yellow solid. LCMS (ESI) m / z [M+H] + = 583.

[0812] Step 4: 4-(2-([4,4'-bipiperidinyl]-1-yl)-2-oxoethoxy)-2-(2,6-dioxopiperidine-3-yl)-1-oxopiperidinyl Preparation of 1,3-dione (I-17)

[0813]

[0814] To a stirred mixture of tert-butyl 1'-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetyl)-[4,4'-bipiperidine]-1-carboxylate (1.36 g, 2.334 mmol, 1.0 equiv) in DCM (40 mL) was added TFA (10 mL) dropwise at room temperature. The resulting mixture was stirred for 2 h and then concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography to yield 4-(2-([4,4'-bipiperidine]-1-yl)-2-oxoethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (I-17, 780 mg, 69.25%) as a yellow solid. LCMS (ESI) m / z [M+H] + = 483.

[0815] In a similar manner as described in the preparation of 1-17, the following intermediates in Table A4 were prepared.

[0816]

[0817] Preparation of (2S,4R)-1-((S)-2-(4-(2-(2-aminoethoxy)ethoxy)butyrylamino)-3,3-dimethylbutyryl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (I-19)

[0818]

[0819] Step 1: Preparation of methyl (2E)-4-(2-[2-[(tert-butoxycarbonyl)amino]ethoxy]ethoxy)but-2-enoate preparation

[0820]

[0821] In 40 mL sealed test tubes, tert-butyl N-[2-(2-hydroxyethoxy)ethyl]carbamate (1.00 g, 4.872 mmol, 1.00 equivalent), (2E)-4-bromobut-2-enoate methyl ester (8.77 g, 48.991 mmol, 10.06 equivalent) and Ag O (3.22 g, 13.895 mmol, 2.85 equivalent) were added at room temperature. The resulting mixture was stirred for 2 days at room temperature under an atmosphere of dry nitrogen. The resulting mixture was filtered, and the filter cake was washed with EtOAc (2 x 10 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with petroleum ether / EtOAc (1:1) to provide methyl (2E)-4-(2-[2-[(tert-butoxycarbonyl)amino]ethoxy]ethoxy)but-2-enoate (380 mg, 25.71%) as a yellow oil. LCMS (ESI) m / z: [M+H] + = 304.

[0822] Step 2: Preparation of 2,2-dimethyl-4-oxo-3,8,11-trioxa-5-azapentadecan-15-oic acid methyl ester

[0823]

[0824] Under nitrogen atmosphere in 50 mL 3-neck round-bottom flask to (2E)-4-(2-[2-[(tert-butoxycarbonyl) amino] ethyoxy] ethyoxy) but-2-enoic acid methyl esters (380.00 mg, 1.253 mmol, 1.00 equivalent) in MeOH (10.00 mL) in solution, add 10% Pd / C (66.65 mg).Mixture is stirred to 4 h under 1 atmospheric pressure of hydrogen.Reactant mixture is passed through pad filtration and under reduced pressure concentrate to provide 2,2-dimethyl-4-oxo-3,8,11-trioxa-5-azapentadecane-15-acid methyl esters (300 mg, 78.43%) as yellow oil, it is not further purified for next step.

[0825] Step 3: Preparation of 2,2-dimethyl-4-oxo-3,8,11-trioxa-5-azapentadecan-15-oic acid

[0826]

[0827] To the 2,2-dimethyl-4-oxo-3,8,11-trioxa-5-azapentadecane-15-acid methyl esters (280.00 mg, 0.917 mmol, 1.00 equivalent) stirred solution in MeOH (2.00 mL) provided in room temperature, dropwise add LiOH (87.83 mg, 3.668 mmol, 4.00 equivalent) in water (2.00 mL).The mixture obtained is stirred at room temperature overnight.Water layer is extracted with EtOAc (2 x 10 mL).Then water layer is acidified to pH 6 with HCl.The mixture obtained is extracted with EtOAc (2 x 10 mL).The organic layer water (30 mL) these merges is washed and through anhydrous Na2SO4 drying. After filtration, the filtrate was concentrated under reduced pressure to give 2,2-dimethyl-4-oxo-3,8,11-trioxa-5-azapentadecan-15-oic acid (100 mg) as a colorless oil, which was used directly in the next step. 1 H NMR (300 MHz, DMSO- d 6) δ 12.06 (s, 1H), 6.78 (d, J = 8.7 Hz, 1H), 3.54 - 3.34 (m, 8H), 3.06 (qd, J = 6.0,3.3 Hz, 2H), 2.25 (t, J = 7.4 Hz, 2H), 1.79 - 1.63 (m, 2H), 1.38 (d, J = 1.3 Hz, 9H).

[0828] Step 4: N-[2-[2-(3-[[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazole-5- 1-yl)phenyl]methyl]carbamoyl]pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamoyl]propyloxy Preparation of tert-butyl]-[(1-methyl)ethoxy]ethyl]carbamate

[0829]

[0830] To a stirred mixture of 2,2-dimethyl-4-oxo-3,8,11-trioxa-5-azapentadecan-15-oic acid (400.00 mg, 1.373 mmol, 1.00 equiv) and HATU (626.44 mg, 1.648 mmol, 1.20 equiv) in DMF (10.00 mL) was added DIEA (212.93 mg, 1.648 mmol, 1.20 equiv) dropwise at room temperature under an atmosphere of dry nitrogen. The resulting mixture was stirred for 30 min. To the above mixture was added (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 (472.92 mg, 1.098 mmol, 0.8 equiv) at room temperature. The resulting mixture was stirred at room temperature overnight and then partitioned between EtOAc (20 mL) and water (50 mL). The organic phase was separated and the aqueous layer was extracted with EtOAc (2 x 10 mL). The combined organic layers were washed with brine (50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with DCM / MeOH (20:1) to afford tert-butyl N-[2-[2-(3-[[(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]propoxy)ethoxy]ethyl]carbamate (300 mg, 31.04%) as a yellow oil. LCMS (ESI) m / z: [M+H] + = 704.

[0831] Step 5: (2S,4R)-1-((S)-2-(4-(2-(2-aminoethoxy)ethoxy) Butyryl amino)-3,3-dimethyl Preparation of 4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (I-19)

[0832]

[0833] To an 8 mL vial was added tert-butyl N-[2-[2-(3-[[(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]propoxy)ethoxy]ethyl]carbamate (300.00 mg, 0.426 mmol, 1.00 equiv) and 4 M HCl in dioxane (3.00 mL) at room temperature and the resulting mixture was stirred for 2 h. The solution was concentrated under reduced pressure to afford (2S,4R)-1-((S)-2-(4-(2-(2-aminoethoxy)ethoxy)butyrylamino)-3,3-dimethylbutyryl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (I-19, 300 mg) as a yellow oil, which was used without further purification. LCMS (ESI) m / z: [M+H] + = 604.

[0834] Preparation of 4-(aminomethyl)-N-(3-chloro-1H-indol-7-yl)benzenesulfonamide (I-20) and 2-amino-N-([4-[(3-chloro-1H-indol-7-yl)sulfamoyl]phenyl]methyl)acetamide (I-21)

[0835]

[0836] Step 1: Preparation of 3-chloro-7-nitro-1H-indole

[0837]

[0838] To a mixture of 7-nitroindole (20.00 g, 123.344 mmol, 1.00 equiv) in THF (100.00 mL) and 0.1 N HCl (1.6 mL) was added NCS (16.47 g, 123.344 mmol, 1 equiv) in one portion. The reaction was stirred at room temperature for 5 h. Water (30 mL) was added. The resulting precipitate was collected by filtration; washed successively with water, MeOH / water (1:1), and isopropyl ether, and dried to provide 3-chloro-7-nitro-1H-indole (24.4 g). LCMS (ESI) m / z: [M+H] + = 198.

[0839] Step 2: Preparation of 3-chloro-1H-indol-7-amine

[0840]

[0841] To a suspension of 3-chloro-7-nitro-1H-indole (24.40 g, 124.116 mmol, 1.00 equivalent) and iron powder (27.73 g, 496.465 mmol, 4 equivalents) in IPA (350.00 mL) was added NH4Cl (53.11 g, 992.929 mmol, 8 equivalents) and water (350.00 mL). The resulting mixture was stirred at 60 ° C for 2 h. The resulting mixture was filtered and the filter cake was washed with EtOH (3 x 500 mL). The filtrate was concentrated under reduced pressure. The resulting mixture was diluted with EtOAc (500 mL) and then basified to pH 8 with saturated NaHCO3 aqueous solution. The resulting mixture was extracted with EtOAc (3 x 30 mL). The combined organic layers were washed with brine (3 x 500 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to provide 3-chloro-1H-indol-7-amine (16 g, 77.37%) as a black solid. LCMS (ESI) m / z: [M+H] + = 167.

[0842] Step 3: Preparation of N-(3-chloro-1H-indol-7-yl)-4-formylbenzenesulfonamide

[0843]

[0844] To 3-chloro-1H-indole-7-amine (1.55 g, 9.303 mmol, 1.00 equivalent) in EtOAc (20.00 mL), add 4-formylbenzenesulfonyl chloride (2.09 g, 10.233 mmol, 1.1 equivalents) and pyridine (1.47 g, 18.606mmol, 2 equivalents). The reaction mixture is stirred at room temperature for 3 h, diluted with EtOAc, and successively washed with 1 N HCl, water, saturated NaHCO aqueous solution and salt water. The organic layer is dried over MgSO, filtered and concentrated under vacuum. The crude product is purified by flash column chromatography using 10% to 60% EtOAc gradient in heptane to obtain N-(3-chloro-1H-indole-7-yl)-4-formylbenzenesulfonamide (1.90 g, 61.01%). LCMS (ESI) m / z: [M+H] + = 335.

[0845] Step 4: Preparation of 4-(aminomethyl)-N-(3-chloro-1H-indol-7-yl)benzenesulfonamide (I-20)

[0846]

[0847] N-(3-Chloro-1H-indol-7-yl)-4-formylbenzenesulfonamide (2.29 g, 6.841 mmol, 1.00 equiv) was dissolved in a saturated solution of ammonium acetate in EtOH (137 mL, prepared by heating 150 mL of EtOH to reflux, then adding ammonium acetate until saturated, followed by 30% aqueous ammonium hydroxide (15.84 mL). After 5 h, NaBH3CN (0.09 mmol, 3 equiv) was added and the reaction mixture was heated to 100°C for 15 min. The residue was purified by reverse phase flash chromatography (C18 silica gel column), eluting with 0-100% acetonitrile in 0.1% formic acid / water over 10 min to provide 4-(aminomethyl)-N-(3-chloro-1H-indol-7-yl)benzenesulfonamide (I-20, 45 mg, 40.05%) as a light yellow solid. LCMS (ESI) m / z: [M+H] + = 336.10.

[0848] Step 5: (2-((4-(N-(3-chloro-1H-indol-7-yl)sulfamoyl)benzyl)amino)-2-oxoethyl)amino Preparation of tert-butyl formate

[0849]

[0850] To a stirred mixture of 4-(aminomethyl)-N-(3-chloro-1H-indol-7-yl)benzenesulfonamide (280.0 mg, 0.834 mmol, 1.00 equiv) and [(tert-butoxycarbonyl)amino]acetic acid (175.28 mg, 1.001 mmol, 1.20 equiv) in DMF (3.00 mL) was added HATU (412.15 mg, 1.084 mmol, 1.30 equiv) and DIEA (323.29 mg, 2.501 mmol, 3.00 equiv) portionwise at room temperature. The mixture was stirred under an atmosphere of dry nitrogen for 2 h. The mixture was concentrated and the residue was purified by reverse phase flash chromatography to give tert-butyl (2-((4-(N-(3-chloro-1H-indol-7-yl)sulfamoyl)benzyl)amino)-2-oxoethyl)carbamate as a white solid (172 mg, 40.17%). LCMS (ESI) m / z: [M+H] + = 493.

[0851] Step 6: 2-amino-N-([4-[(3-chloro-1H-indol-7-yl)sulfamoyl]phenyl]methyl)acetamide (I- 21) Preparation

[0852]

[0853] A mixture of tert-butyl (2-((4-(N-(3-chloro-1H-indol-7-yl)sulfamoyl)benzyl)amino)-2-oxoethyl)carbamate (162.00 mg, 0.329 mmol, 1.00 equiv) and TFA (1.00 mL, 13.463 mmol, 40.97 equiv) in DCM (4.00 mL) was stirred at room temperature under an atmosphere of dry nitrogen for 2 h. The solution was concentrated, and the residue was purified by reverse phase flash chromatography to afford 2-amino-N-([4-[(3-chloro-1H-indol-7-yl)sulfamoyl]phenyl]methyl)acetamide (I-21, 112 mg, 83.28%) as a white solid. LCMS (ESI) m / z: [M+H] + = 393.

[0854] Preparation of 5-[(8-aminooctyl)amino]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione TFA (I-22)

[0855]

[0856] Step 1: Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-fluoro-2,3-dihydro-1H-isoindole-1,3-dione

[0857]

[0858] To a stirred mixture of 5-fluoro-1,3-dihydro-2-benzofuran-1,3-dione (5.6 g, 33.713 mmol, 1.00 equiv) in AcOH (60 mL) was added 2,6-dioxopiperidin-3-ammonium chloride (5.55 g, 33.713 mmol, 1.00 equiv) and sodium acetate (5.53 g, 67.426 mmol, 2.00 equiv) at 120°C under an atmosphere of dry nitrogen. After 14 h, the reaction mixture was concentrated under reduced pressure to remove most of the acetic acid. The residue was poured into water (100 mL) and stirred for 10 min. The mixture was filtered. The filter cake was washed with water and dried. This produced the title compound (8.03 g, 81.92%) as a pink solid. LCMS (ESI) m / z: [M+H] + = 277.

[0859] Step 2: N-(8-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindole- Preparation of tert-butyl 5-[(5-yl]amino]octyl)carbamate

[0860]

[0861] To a stirred mixture of 2-(2,6-dioxopiperidin-3-yl)-5-fluoro-2,3-dihydro-1H-isoindole-1,3-dione (5.00 g, 18.101 mmol, 1.00 equiv) and tert-butyl N-(8-aminooctyl)carbamate (6.64 g, 27.152 mmol, 1.50 equiv) in NMP (50.00 mL) at 90°C was added DIEA (7.02 g, 54.304 mmol, 3.00 equiv). After 3 h, water (100 mL) was added to the mixture, followed by extraction with EtOAc (200 mL x 3). The residue was purified by reverse-phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, ACN in water, 0% to 50% gradient; detector, UV 254 nm. This gave the title compound as a yellow solid (3.50 g, 36.69%). LCMS (ESI) m / z: [M+H] + = 501.

[0862] Step 3: 5-[(8-aminooctyl)amino]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione tris(II) Preparation of fluoroacetate (I-22)

[0863]

[0864] To a stirred mixture of tert-butyl N-(8-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]amino]octyl)carbamate (3.85 g, 7.691 mmol, 1.00 equiv) in DCM (9.00 mL) was added TFA (3.00 mL) at room temperature. The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by CombiFlash to afford I-22 (2.22 g, 56.20%) as a yellow solid. 1H NMR (400 MHz, DMSO) δ 11.08 (s, 1H), 7.80 - 7.72 (m, 1H), 7.72 -7.67 (m, 2H), 7.55 (dd, 1H), 7.14 (s, 1H), 6.95 (d, 1H), 6.88 - 6.77 (m, 1H),5.08 - 4.98 (m, 1H), 3.21 - 3.05 (m, 1H), 2.95 - 2.75 (m, 2H), 2.66 - 2.51(m, 2H), 2.12 - 1.94 (m, 1H), 1.63 - 1.49 (m, 3H), 1.45 - 1.31 (m, 4H). LCMS (ESI) m / z: [M+H] + = 401.21.

[0865] In a similar manner as described in the preparation of 1-22, the following intermediates in Table A5 were prepared.

[0866]

[0867]

[0868] Preparation of 4-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-4-yl]oxy]butanoic acid (I-29)

[0869]

[0870] Step 1: tert-4-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]butanoic acid Preparation of butyl ester

[0871]

[0872] To a solution of 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindole-1,3-dione (2.00 g, 7.293 mmol, 1.00 equiv) and tert-butyl 4-bromobutyrate (1.95 g, 8.752 mmol, 1.2 equiv) in DMF (10.00 mL) was added KI (0.12 g, 0.729 mmol, 0.1 equiv) and KHCO (1.10 g, 10.940 mmol, 1.5 equiv). 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 x 3). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by flash C18 chromatography eluting with a gradient of 0-32% ACN in water to give the title product as an off-white solid (1.5 g, 49.39%). LCMS (ESI) m / z [M+H] + = 417.

[0873] Step 2: 4-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindole-4- Preparation of 1-[3-[[4-(2-amino-3-methyl-1-oxy-butyric acid)]]-1-[3-amino-3-methyl-1-oxy-butyric acid ...

[0874]

[0875] To a stirred solution of tert-butyl 4-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]butanoate (450 mg, 1.081 mmol, 1 equiv) in DCM (5 mL) was added TFA (1 mL). The resulting solution was stirred at 25 ° C for 2 h. The resulting mixture was concentrated. This provided I-29 (360 mg, 92.46%) as a white solid. 1 H NMR (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.

[0876] In a similar manner as described in the preparation of intermediate 1-29, the following intermediates in Table A6 were prepared.

[0877]

[0878] Preparation of 3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)propionic acid (I-34)

[0879]

[0880] Step 1: 3-(2-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino Preparation of tert-butyl (1,2-dimethyl)ethoxy)ethoxy)propionate

[0881]

[0882] To a solution of 2-(2,6-dioxopiperidin-3-yl)-4-fluoro-2,3-dihydro-1H-isoindole-1,3-dione (1.00 g, 3.620 mmol, 1.00 equivalent) in NMP (10.00 mL) was added tert-butyl 3-[2-(2-aminoethoxy)ethoxy]propanoate (929.10 mg, 3.982 mmol, 1.10 equivalent). The resulting mixture was stirred at 90 ° C overnight. The mixture was cooled to room temperature. The resulting mixture was diluted with EtOAc (30 mL). The organic layer was washed with water (10 mL x 5) and then washed with brine (20 mL). The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with petroleum ether / EtOAc (from 5:1 to 1:1) to afford tert-butyl 3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)propanoate (1.14 g, 64.33%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 490.

[0883] Step 2: 3-(2-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino Preparation of 2-(4-(2-ethoxy)ethoxy)propionic acid (I-34)

[0884]

[0885] To a stirred solution of tert-butyl 3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)propanoate (1.14 g, 2.329 mmol, 1.00 equiv) in DCM (10.00 mL) was added TFA (0.52 mL, 4.551 mmol, 3.00 equiv) dropwise at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under vacuum and the residue was purified by reverse phase flash chromatography using the following conditions: column, C18 spherical column; mobile phase, ACN in water, 0% to 100% gradient over 50 min; 70 mL / min detector, UV 254 nm to afford 1-34 (896 mg, 70.28%) as a yellow solid. 1 H NMR (400 MHz, DMSO) 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.

[0886] The following intermediates in Table A7 were prepared in a similar manner as described in the preparation of intermediate 1-34.

[0887]

[0888]

[0889] Preparation of 4-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-5-yl)amino)butyric acid (K-19)

[0890]

[0891] Step 1: 4-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazine- Preparation of tert-butyl 5-aminobutyrate

[0892]

[0893] Into an 8-mL sealed test tube was placed 3-(5-fluoro-4-oxo-1,2,3-benzotriazin-3-yl)piperidine-2,6-dione (100.00 mg, 0.362 mmol, 1.00 equiv), NMP (1.50 mL), tert-butyl 4-aminobutyrate (86.47 mg, 0.543 mmol, 1.5 equiv), and DIEA (140.37 mg, 1.086 mmol, 3 equiv). The resulting solution was stirred at 90°C for 5 h. The reaction mixture was purified by reverse-phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, MeCN in water, gradient from 10% to 50% over 10 min; detection, UV 254 nm. This resulted in 100 mg (66.49%) of tert-butyl 4-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-5-yl)amino)butanoate as a yellow solid. LCMS (ESI) m / z: [M+H] + =416.

[0894] Step 2: 4-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazine- Preparation of 5-aminobutyric acid (K-19)

[0895]

[0896] In an 8-mL sealed test tube was placed tert-butyl 4-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-5-yl)amino)butanoate (90.00 mg, 0.217 mmol, 1.00 equiv), DCM (5.00 mL) and TFA (0.50 mL). The resulting solution was stirred at room temperature for 30 min and then concentrated under reduced pressure to afford 4-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-5-yl)amino)butanoic acid (70 mg, quantitative) as a light yellow crude solid. The crude product was used in the next step without further purification. LCMS (ESI) m / z: [M+H] + = 360.

[0897] The following intermediates in Table A8 were prepared in a similar manner as described in the preparation of intermediate K-19.

[0898]

[0899] Preparation of 5-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-6-yl)amino)pentanoic acid (K-22)

[0900]

[0901] Step 1: 5-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazol-2 ... Preparation of methyl 6-oxazine-6-yl)amino)pentanoate

[0902]

[0903] To a stirred solution of 3-(6-amino-4-oxo-1,2,3-benzotriazine-3-yl)piperidine-2,6-dione (120.00 mg, 0.439 mmol, 1.00 equiv) and methyl 5-oxopentanoate (57.15 mg, 0.439 mmol, 1.00 equiv) in MeOH (2.00 mL) was added AcOH (0.20 mL) portionwise at room temperature. The resulting mixture was stirred at 50 ° C for 2 h. To the above mixture was added NaBH3CN (82.79 mg, 1.317 mmol, 3.00 equiv) portionwise over 1 min. The resulting mixture was stirred at 50 ° C for another 2 h. The resulting mixture was diluted with water (20 mL). The aqueous layer was extracted with CHCl (4x20 mL). The resulting mixture was concentrated under reduced pressure to afford methyl 5-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-6-yl)amino)pentanoate, which was used in the next step without further purification. LCMS (ESI) m / z [M+H] + =388.15.

[0904] Step 2: 5-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazol-2 ... Preparation of 6-oxazin-6-yl)amino)pentanoic acid (K-22)

[0905]

[0906] To a 25 mL vial was added methyl 5-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-6-yl)amino)pentanoate (120.00 mg, 0.310 mmol, 1.00 equiv) and HCl (6M, 5.00 mL, 0.027 mmol, 5.00 equiv) at room temperature. The resulting mixture was stirred at room temperature under an air atmosphere for 1 h and then concentrated under reduced pressure to provide crude 5-((3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrobenzo[d][1,2,3]triazin-6-yl)amino)pentanoic acid, which was used in the next step without further purification. LCMS (ESI) m / z [M+H] + =374.10.

[0907] Preparation of 2-[(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]acetylamino)methyl]cyclopropane-1-carboxylic acid (I-42)

[0908]

[0909] Step 1: tert-2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]acetic acid Preparation of butyl ester

[0910]

[0911] To a stirred solution of 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindole-1,3-dione (5.50 g, 20.056 mmol, 1.00 equiv) and tert-butyl 2-bromoacetate (3.91 g, 20.056 mmol, 1.00 equiv) in DMF (15.00 mL) was added KCO (8.32 g, 60.168 mmol, 3 equiv). The resulting mixture was stirred at room temperature overnight, then taken up in water, extracted with EtOAc (3 x 200 mL), and concentrated. The residue was purified by reverse-phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, ACN in water, 10% to 50% gradient over 10 minutes; detector, UV 254 nm. This resulted in tert-butyl 2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]acetate (3.2 g, 45.19%) as an off-white solid. LCMS (ESI) m / z: [M+H] + = 389.

[0912] Step 2: Preparation of [[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]acetic acid Preparation

[0913]

[0914] Tert-butyl 2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]acetate (3.20 g, 8.239 mmol, 1.00 equiv) and dry HCl in dioxane (15.00 mL, 493.678 mmol, 59.92 equiv) were stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. This afforded the title compound (1.12 g) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 289.

[0915] Step 3: 2-[(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]acetyl Preparation of methyl [amino]methyl]cyclopropane-1-carboxylate

[0916]

[0917] To a stirred solution of [[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]acetic acid (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) was added HATU (1150.07 mg, 3.025 mmol, 1.50 equiv) and DIEA (781.84 mg, 6.049 mmol, 3.00 equiv) dropwise at room temperature over 2 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, ACN in water, 10% to 50% gradient over 10 min; detector, UV 254 nm. This yielded methyl 2-[(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoindole-5-yl]oxy]acetamido)methyl]cyclopropane-1-carboxylate (778.6 mg, 78.37%) as a yellow oil. LCMS (ESI) m / z: [M+H] + = 444.

[0918] Step 4: 2-[(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]ethyl Preparation of [4-(4-Acylamino)methyl]cyclopropane-1-carboxylic acid (I-42)

[0919]

[0920] A mixture of methyl 2-[(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]acetamido)methyl]cyclopropane-1-carboxylate (763.90 mg, 1.00 equiv) and 4 N dry HCl in dioxane (5.00 mL) was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, ACN in water, 10% to 50% gradient over 10 min; detector, UV 254 nm. This afforded I-42 (338.2 mg) as a yellow solid. 1H NMR (400 MHz, DMSO) δ12.17 (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.

[0921] Preparation of 2-[(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]acetylamino)methyl]cyclopropane-1-carboxylic acid (I-43)

[0922]

[0923] In a similar manner as described in the preparation of 1-42, 1-43 was prepared as a white solid (423 mg, 35.74%). 1 H NMR (300 MHz, DMSO) 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.

[0924] 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-44)

[0925]

[0926] To a stirred solution of suberic acid (2.02 g, 11.596 mmol, 4.99 equiv) in DCM (25.00 mL) and THF (25.00 mL) was added (2S,4R)-1-(2-amino-3,3-dimethyl-butyryl)-4-hydroxy-N-[[4-(4-methylthiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (1.00 g, 2.323 mmol, 1.00 equiv) and TEA (822.55 mg, 8.129 mmol, 3.50 equiv) and HOAt (347.73 mg, 2.555 mmol, 1.10 equiv) and EDCI (489.75 mg, 2.555 mmol, 1.10 equiv) at 0° C. The resulting solution was stirred at 0° C. for 2 h. The reaction was confirmed to be complete by LCMS, and the resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography to provide I-44 (900 mg, 66.04%) as a white solid. LCMS (ESI) m / z: [M+H] + = 587.

[0927] 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-45)

[0928]

[0929] Step 1: 3-(2-(3-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)amino (formyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)ethoxy)propanoic acid Preparation of ethyl ester

[0930]

[0931] To a solution of (2S,4R)-1-(2-amino-3,3-dimethyl-butyryl)-4-hydroxy-N-[[4-(4-methylthiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (500 mg, 1.07 mmol, HCl), 3-[2-(3-ethoxy-3-oxo-propoxy)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. The mixture was concentrated under reduced pressure to yield a residue. The residue was purified by reverse phase HPLC (ACN / 0.1% aqueous formic acid). The solution was lyophilized to produce 3-(2-(3-(((S)-1-((2S, 4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutane-2-yl)amino)-3-oxopropoxy)ethoxy)ethyl propionate (500 mg, 70.00%) as a yellow oil. LCMS (ESI) m / z: [M+H] + = 647.6.

[0932] Step 2: 3-(2-(3-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)amino (formyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)ethoxy)propanoic acid Preparation of (I-45)

[0933]

[0934] To the resulting solution of ethyl 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)propanoate (500 mg, 773.05 µm) in EtOH (5 mL) was added a solution of NaOH (77.30 mg, 1.93 mmol) in water (1 mL). The mixture was stirred at 20°C for 1 h. The solution was diluted with water (30 mL), adjusted to pH 6 with 1N HCl, and extracted with EtOAc (30 mL x 3). The combined organic layers were dried over NaSO and concentrated to give I-45 (470 mg, 95.02%) as a brown solid, which was used in the next step without purification. LCMS (ESI) m / z: [M+Na] + =641.2.

[0935] In a similar manner as described in the preparation of 1-45, the following intermediates in Table A9 were prepared.

[0936]

[0937]

[0938]

[0939] 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-azapentadecan-1-oic acid (I-57)

[0940]

[0941] Step 1: Preparation of 2-[2-[2-(2-benzyloxy-2-oxo-ethoxy)ethoxy]ethoxy]acetic acid

[0942]

[0943] To a mixture of 2-[2-[2-(carboxymethoxy)ethoxy]ethoxy]acetic acid (3 g, 13.50 mmol) and TEA (3.51 g, 34.71 mmol, 4.83 mL) in acetone (20 mL) was added bromomethylbenzene (2.42 g, 14.18 mmol, 1.68 mL) dropwise at 0 ° C. The mixture was stirred at 20 ° C for 16 h. A thick precipitate was formed, which was filtered out and the filter cake was washed with acetone (10 mL). The filtrate was concentrated and the residue was dissolved in water (300 mL). The mixture was extracted with EtOAc (50 mL x 3) and then treated with 2 N HCl to a pH of 3-5. The mixture was extracted with EtOAc (50 mL x 3) and the organic layers were combined and washed with brine (50 mL), then dried over Na2SO4, filtered and concentrated under vacuum to give 2-[2-[2-(2-benzyloxy-2-oxo-ethoxy)ethoxy]ethoxy]acetic acid (1.4 g, 33.20%) as a yellow oil, which was used in the next step without further purification. 1 H NMR (400 MHz, CDCl3) 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.

[0944] Step 2: (S)-13-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidone Preparation of benzyl (1-oxo-14,14-dimethyl)-11-oxo-3,6,9-trioxa-12-azapentadecan-1-olate

[0945]

[0946] To a solution of 2-[2-[2-(2-benzyloxy-2-oxo-ethoxy)ethoxy]ethoxy]acetic acid (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). (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) was added. The mixture was stirred at 30° C. for 2 h. The mixture was concentrated under vacuum to give a yellow solid, which was purified by reverse phase flash chromatography (ACN / 0.1% aqueous formic acid) to yield (S)-benzyl 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-azapentadecan-1-oate (1.2 g, 1.66 mmol, 71.28%) as a yellow oil. 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.

[0947] Step 3: (S)-13-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidone Preparation of (1-oxo-3,6,9-trioxa-12-azapentadecan-1-ol)-14,14-dimethyl-11-oxo-3,6,9-trioxa-12-azapentadecan-1-ol (I-57)

[0948]

[0949] To a mixture 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-azapentadecan-1-oic acid benzyl ester (1.1 g, 1.52 mmol) in MeOH (20 mL) was added 10% Pd / C (500 mg). The mixture was stirred at 25 ° C for 12 h under 15 psi of hydrogen. The mixture was then stirred at 40 ° C for 8 h. The mixture was filtered to remove the Pd / C and the filtrate was concentrated under vacuum. The residue was purified by reverse phase chromatography to yield I-57 (560 mg, 58.14%) 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. Chiral SFC: AD-3-MeOH+ACN (DEA)-40-3ML-35T.lcm, Rt= 0.419 min, ee > 100%.

[0950] In a similar manner as described in the preparation of 1-57, the following intermediates in Table A10 were prepared.

[0951]

[0952] Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-(piperazin-1-yl)isoindole-1,3-dione; formic acid (I-59) and 5-[4-(2,2-diethoxyethyl)piperazin-1-yl]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (I-60)

[0953]

[0954] Step 1: tert-4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]piperazine-1-carboxylic acid Preparation of butyl ester

[0955]

[0956] To a solution of 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindole-1,3-dione (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) at room temperature. The resulting mixture was stirred at 90 ° C for 5 h. The reaction was quenched at room temperature by adding water (50 mL). The resulting mixture was extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (3 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. This resulted in tert-butyl 4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]piperazine-1-carboxylate (7.32 g, 84.62%) as a light yellow solid. LCMS (ESI) m / z: [M+H] + = 443.

[0957] Step 2: 2-(2,6-dioxopiperidin-3-yl)-5-(piperazin-1-yl)isoindole-1,3-dionecarboxylic acid (I-59) Preparation

[0958]

[0959] To a solution of tert-butyl 4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]piperazine-1-carboxylate (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. The residue was purified by reverse phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, 0.1% aqueous formic acid / ACN, gradient from 0% to 100% over 45 min; detector, UV 254 nm. This produced I-59 (786.8 mg, 99.41%) as a yellow solid. LCMS (ESI) m / z: [M+H] + =343.

[0960] Step 3: 5-[4-(2,2-diethoxyethyl)piperazin-1-yl]-2-(2,6-dioxopiperidin-3-yl)isoindole Preparation of Indole-1,3-dione (I-60)

[0961]

[0962] To a solution of I-59 (1.15 g, 3.359 mmol, 1.00 equiv) and 2-bromo-1,1-diethoxyethane (0.66 g, 3.359 mmol, 1 equiv) in DMF (20.00 mL) was added DIEA (1.30 g, 10.077 mmol, 3 equiv). The resulting mixture was stirred at 80 °C for 12 h. The residue was purified by reverse phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, ACN 0.1% formic acid in water, 0% to 100% gradient over 45 min; detector, UV 254 nm. This produced I-60 (987 mg, 64.08%) as a white solid; 1 H NMR (300 MHz, DMSO) δ 11.09 (s,1H), 7.68 (d, 1H), 7.34 (d, 1H), 7.26 (dd, 1H), 5.08 (dd, 1H), 4.64 (t, 1H),3.67 - 3.58 (m, 2H), 3.57 - 3.38 (m, 6H), 3.33 (s, 1H), 2.98 - 2.80 (m, 1H), 2.66 - 2.59 (m, 4H), 2.56 (s, 2H), 2.48 (s, 1H), 2.09 - 1.96 (m, 1H), 1.13(t, 6H). LCMS (ESI) m / z: [M+H] + = 459.30.

[0963] Preparation of 1-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]ethyl)piperidine-4-carboxylic acid (I-61)

[0964]

[0965] Step 1: Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-(prop-2-en-1-yloxy)isoindole-1,3-dione preparation

[0966]

[0967] To a solution of 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindole-1,3-dione (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 KHCO₃ (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 x 50 mL). The combined organic layers were washed with brine (3 x 60 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 hexane / EtOAc (1:1) to provide 2-(2,6-dioxopiperidin-3-yl)-5-(prop-2-en-1-yloxy)isoindole-1,3-dione (6.7 g, crude) as a yellow-green solid. LCMS (ESI) m / z: [M+H] + = 315.

[0968] Step 2: 2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]acetaldehyde preparation

[0969]

[0970] To a solution of 2-(2,6-dioxopiperidin-3-yl)-5-(prop-2-en-1-yloxy)isoindole-1,3-dione (3.14 g, 9.991 mmol, 1.00 equiv) in dioxane (30.00 mL) was added NaIO (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 KOsO dihydrate (0.37 g, 0.999 mmol, 0.1 equiv) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. The reaction was quenched with water at room temperature and the resulting mixture was extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (3 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. This produced 2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]acetaldehyde (1.83 g, 57.92%) as a light brown solid. LCMS (ESI) m / z: [M+H] + = 317.

[0971] Step 3: 1-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]ethyl Preparation of tert-butyl (4-methyl)piperidine-4-carboxylate

[0972]

[0973] To a solution of 2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]acetaldehyde (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. The residue was purified by reverse-phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, ACN / 0.1% formic acid in water, gradient from 0% to 100% over 45 min; detector, UV 254 nm. This resulted in tert-butyl 1-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]ethyl)piperidine-4-carboxylate (1.16 g, 41.29%) as an off-white solid. LCMS (ESI) m / z: [M+H] + = 401.

[0974] Step 4: 1-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]ethyl Preparation of 4-(2-amino)piperidine-4-carboxylic acid (I-61)

[0975]

[0976] To a solution of tert-butyl 1-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]ethyl)piperidine-4-carboxylate (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 resulting mixture was stirred at room temperature for 5 h. The residue was purified by reverse phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, ACN / 0.1% aqueous formic acid, gradient from 0% to 100% over 45 min; detector, UV 254 nm. This yielded I-61 (845 mg, 73.42%) as a white solid. 1HNMR (300 MHz, DMSO) 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.

[0977] Preparation of 4-(3-aminopropoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (K-32)

[0978]

[0979] Step 1: Preparation of tert-butyl N-[3-[(4-methylbenzenesulfonyl)oxy]propyl]carbamate

[0980]

[0981] To a solution of tert-butyl N-(3-hydroxypropyl)carbamate (1.00 g, 5.707 mmol, 1.00 equiv) in DCM (40.00 mL) was added TsCl (1.32 g, 6.924 mmol, 1.21 equiv) and TEA (2.00 mL). The reaction mixture was stirred at room temperature overnight. The resulting mixture was neutralized to pH 8 with aqueous NaHCO3 solution. The organic layer was separated and washed with brine (20 mL) and then dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with PE / EtOAc (5:1) to provide tert-butyl N-[3-[(4-methylbenzenesulfonyl)oxy]propyl]carbamate (750 mg, 39.90%) as a colorless oil. LCMS (ESI) m / z: [M+H] + = 330.

[0982] Step 2: N-(3-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]propane Preparation of tert-butyl (2-(2-methyl-1,2-dimethylamino)carbamate.

[0983]

[0984] Prepare a suspension of tert-butyl N-[3-[(4-methylbenzenesulfonyl)oxy]propyl]carbamate (225 mg), 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindole-1,3-dione (300 mg, 1.00 equiv), and NaCO (200 mg) in DMF (2 mL). The mixture was stirred at 80°C for 2 h. The resulting mixture was diluted with DCM (10 mL) and washed with HO (2 x 10 mL). The organic layer was separated and dried over NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (eluting with PE / EtOAc 1:1) to afford tert-butyl N-(3-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]propyl)carbamate (70 mg, 14.83%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 432.

[0985] Step 3: 4-(3-aminopropoxy)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (K-32) Preparation

[0986]

[0987] To a solution of tert-butyl N-(3-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]propyl)carbamate (70.00 mg, 0.162 mmol, 1.00 equiv) in DCM (5.00 mL) was added TFA (1.00 mL). The mixture was stirred at room temperature for 0.5 h. The resulting mixture was concentrated under reduced pressure. The crude product, 4-(3-aminopropoxy)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (70 mg), was used in the next step without further purification. LCMS (ESI) m / z: [M+H] + = 332.

[0988] In a similar manner as described in the preparation of K-32, the following intermediates in Table A11 were prepared.

[0989]

[0990] Preparation of 4-(2-aminoethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (K-35)

[0991]

[0992] Step 1: N-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]ethyl Preparation of tert-butyl (2-(2-methyl-1,2-d)carbamate

[0993]

[0994] To a stirred mixture of 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindole-1,3-dione (500.00 mg, 1.823 mmol, 1.00 equiv) and tert-butyl 1,2,3-oxathiazolidine-3-carboxylate 2,2-dioxide (407.03 mg, 1.823 mmol, 1.00 equiv) in DMF (5.00 mL) was added DIEA (471.29 mg, 3.647 mmol, 2.00 equiv) dropwise at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 80 ° C. under a nitrogen atmosphere overnight. The resulting mixture was diluted with water (10 mL) and then extracted with EtOAc (3 x 10 mL). The organic layers were combined, washed with brine (10 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography to provide tert-butyl N-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]ethyl)carbamate (400 mg, 52.56%) as an off-white solid. LCMS (ESI) m / z [M+H] + =418.

[0995] Step 2: 4-(2-aminoethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (K-35) Preparation

[0996]

[0997] To a stirred solution of tert-butyl N-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]ethyl)carbamate (150.00 mg, 0.359 mmol, 1.00 equiv) in DCM (4.00 mL) was added TFA (0.8 mL) dropwise at room temperature under nitrogen atmosphere. The resulting mixture was stirred at room temperature under nitrogen atmosphere for 1 h and then concentrated under reduced pressure to afford intermediate 3 (100 mg) as a colorless oil. The crude product, 4-(2-aminoethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione, was used in the next step without further purification. LCMS (ESI) m / z [M+H] + =318.

[0998] Preparation of 3-[1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]azetidin-3-yl]propanal (K-36)

[0999]

[1000] Step 1: Preparation of 3-(azetidin-3-yl)propan-1-ol.

[1001]

[1002] To a solution of tert-butyl 3-(3-hydroxypropyl)azetidine-1-carboxylate (120.00 mg, 0.557 mmol, 1.00 equiv) in DCM (2.00 mL) was added TFA (0.50 mL, 6.732 mmol, 12.08 equiv). The mixture was stirred at room temperature for 0.5 h. The reaction mixture was concentrated to provide 3-(azetidin-3-yl)propan-1-ol (70 mg, crude) as a yellow oil. LCMS (ESI) m / z [M+H] + =116.

[1003] Step 2: 2-(2,6-dioxopiperidin-3-yl)-4-[3-(3-hydroxypropyl)azetidin-1-yl]isoindole Preparation of Indole-1,3-Dione

[1004]

[1005] To a solution of 3-(azetidin-3-yl)propane-1-ol (70 mg, 1.337 mmol, 1.00 equiv) and 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindole-1,3-dione (155.12 mg, 0.562 mmol, 1.00 equiv) in NMP (1 mL) was added DIEA (1 mL). The reaction mixture was stirred at 80 ° C for 2 h. The mixture was diluted with DCM (10 mL) and washed with brine (2 x 10 mL). The organic layer was dried over Na2SO4 and filtered. The filtrate was concentrated. The residue was purified by preparative TLC (eluting with 1:1 PE / EtOAc) to provide 2-(2,6-dioxopiperidin-3-yl)-4-[3-(3-hydroxypropyl)azetidin-1-yl]isoindole-1,3-dione (100 mg, 47.95%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 372.

[1006] Step 3: 3-[1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]azetidine Preparation of [alkyl-3-yl] propanal (intermediate K-36)

[1007]

[1008] 2-(2,6-dioxopiperidin-3-yl)-4-[3-(3-hydroxypropyl)azetidin-1-yl]isoindole-1,3-dione (100.00 mg, 0.269 mmol, 1.00 equiv) and Dess-Martin periodinane (114.20 mg, 0.269 mmol, 1.00 equiv) were suspended in DCM (4.00 mL). The mixture was stirred at room temperature overnight. The mixture was purified by preparative TLC (eluting with 1:1 PE / EtOAc) to provide 3-[1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]azetidin-3-yl]propanal (60 mg, 60.33%) as a yellow solid. LCMS (ESI) m / z [M+H] + =370.

[1009] In a similar manner as described in the preparation of K-36, the following intermediates in Table A12 were prepared.

[1010]

[1011] Preparation of 5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)valeraldehyde (K-39)

[1012]

[1013] Step 1: 4-(4-(1,3-dioxolan-2-yl)butoxy)-2-(2,6-dioxopiperidin-3-yl)iso Preparation of indoline-1,3-dione

[1014]

[1015] A solution of 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindole-1,3-dione (500.00 mg, 1.823 mmol, 1.00 equiv), 2-(4-bromobutyl)-1,3-dioxolane (457.46 mg, 2.188 mmol, 1.20 equiv), KI (60.53 mg, 0.365 mmol, 0.20 equiv), and NaHCO (306.33 mg, 3.646 mmol, 2.00 equiv) in DMF (5.00 mL) was stirred at 70° C. under a nitrogen atmosphere for 24 h. The resulting mixture was diluted with ethyl acetate (20 mL) and then washed with saturated NaCl (3 x 20 mL). The organic layer was dried over anhydrous NaSO and then filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography eluting with DCM / MeOH = 100 / 1 to provide 4-(4-(1,3-dioxolan-2-yl)butoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (600 mg, 81.78%) as a light yellow solid. LCMS (ESI) m / z [M+H] + = 403.

[1016] Step 2: 5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)pentanal Preparation of (K-39)

[1017]

[1018] A solution of 4-(4-(1,3-dioxolan-2-yl)butoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (170.00 mg, 0.422 mmol, 1.00 equiv) in HCl (4.00 mL) / THF (4.00 mL) was stirred at room temperature for 10 h. The mixture was neutralized with saturated NaHCO3 to pH 7. The resulting mixture was extracted with ethyl acetate (3 x 20 mL). The combined organic layers were washed with saturated brine (20 mL) and then dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to provide crude 5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)oxy)pentanal (140 mg, 92.48%), which was used in the next step without further purification. LCMS (ESI) m / z [M+H] + = 359.

[1019] Preparation of 2-(4-(3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)piperazin-1-yl)acetic acid (I-62)

[1020]

[1021] Step 1: Preparation of ethyl 2-[4-(3-amino-6-chloropyridazin-4-yl)piperazin-1-yl]acetate

[1022]

[1023] To a stirred mixture of 4-bromo-6-chloropyridazin-3-amine (420.00 mg, 2.015 mmol, 1.00 equiv) and ethyl 2-(piperazin-1-yl)acetate (381.74 mg, 2.216 mmol, 1.10 equiv) in DMF (3.00 mL) was added DIEA (1.30 g, 10.059 mmol, 4.99 equiv) portionwise under an atmosphere of dry nitrogen at 120 ° C over 12 h. The mixture was cooled to room temperature. The residue was purified by reverse phase flash chromatography to provide ethyl 2-[4-(3-amino-6-chloropyridazin-4-yl)piperazin-1-yl]acetate (340 mg, 50.66%) as a light yellow solid. LCMS (ESI) m / z: [M+H] + = 299.76.

[1024] Step 2: Preparation of ethyl 2-[4-[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]piperazin-1-yl]acetate

[1025]

[1026] To a stirred solution of ethyl 2-[4-(3-amino-6-chloropyridazin-4-yl)piperazin-1-yl]acetate (340.00 mg, 1.134 mmol, 1.00 equiv) and 2-hydroxyphenylboronic acid (234.67 mg, 1.701 mmol, 1.50 equiv) in dioxane:water (5 mL, 4:1) was added potassium carbonate (391.90 mg, 2.836 mmol, 2.50 equiv) and XPhos Pd G3 (192.02 mg, 0.227 mmol, 0.20 equiv) portionwise over 2 h at 100 °C under an atmosphere of dry nitrogen. The residue was purified by reverse phase flash chromatography to give ethyl 2-[4-[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]piperazin-1-yl]acetate (253 mg, 58.04%) as an off-white solid. LCMS (ESI) m / z: [M+H] + =357.41.

[1027] Step 3: Preparation of 2-(4-(3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)piperazin-1-yl)acetic acid (I-62) Preparation

[1028]

[1029] To a solution of ethyl 2-[4-[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]piperazin-1-yl]acetate (250.00 mg, 0.699 mmol, 1.00 equiv) was added portionwise a solution of LiOH (348.92 mg, 14.570 mmol, 20.83 equiv) in 1:1 THF / water (10 mL) at room temperature over 2 h. The residue was purified by reverse phase flash chromatography to afford 2-(4-(3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)piperazin-1-yl)acetic acid (I-62, 192 mg, 76.67%) as an off-white solid. LCMS (ESI) m / z: [M+H] + = 329.36.

[1030] 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 hydrochloride (I-63)

[1031]

[1032] Step 1: tert-2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]acetic acid Preparation of butyl ester

[1033]

[1034] To a solution of 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindole-1,3-dione (5.50 g, 20.056 mmol, 1.00 equiv) in DMF (65.0 mL) were added tert-butyl 2-bromoacetate (3.91 g, 20.056 mmol, 1 equiv) and KCO (8.32 g, 60.168 mmol, 3 equiv). The resulting solution was stirred at room temperature for 12 h. The solution was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with petroleum ether / EtOAc (1:1) to provide tert-butyl 2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]acetate (5.2 g, 66.76%) as a white solid. LCMS (ESI) m / z: [M+H] + = 389.

[1035] Step 2: Preparation of [[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]acetic acid Preparation

[1036]

[1037] A solution of tert-butyl 2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]acetate (5.32 g, 13.698 mmol, 1.00 equiv) in 4N HCl (in dioxane) (50.0 mL, 1645.594 mmol, 120.13 equiv) was stirred at room temperature for 12 h. The solution was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography (C18 silica gel column), eluting with 0-100% ACN in water over 35 minutes and detecting at UV 254 nm. This yielded [[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]acetic acid (4.66 g, 100%) as a white solid. LCMS (ESI) m / z: [M+H] + = 333.

[1038] Step 3: N-(2-[[2-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoindole-4-yl]oxy Preparation of tert-butyl [(methyl)amino]ethyl](methyl)amino]ethyl)carbamate

[1039]

[1040] To a solution of [[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]acetic acid (926.00 mg, 2.787 mmol, 1.00 equiv) in DCM (35.00 mL) was added tert-butyl N-[2-[(2-aminoethyl)(methyl)amino]ethyl]carbamate (908.45 mg, 4.180 mmol, 1.5 equiv), HATU (1.59 g, 4.180 mmol, 1.5 equiv) and DIEA (1.08 g, 8.361 mmol, 3 equiv). The resulting solution was stirred at room temperature for 3 h. The residue was purified by silica gel column chromatography eluting with petroleum ether / EtOAc (1:1) to afford tert-butyl N-(2-[[2-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]acetylamino)ethyl](methyl)amino]ethyl)carbamate (1.527 g, crude) as a light yellow solid. LCMS (ESI) m / z: [M+H] + =532.

[1041] Step 4: N-[2-[(2-aminoethyl)(methyl)amino]ethyl]-2-[[2-(2,6-dioxopiperidine-3- Preparation of [1,3-dioxo-2,3-dihydro-1H-isoindol-4-yl]oxy]acetamide hydrochloride (I-63)

[1042]

[1043] A solution of tert-butyl N-(2-[[2-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]acetamido)ethyl](methyl)amino]ethyl)carbamate (600.00 mg, 1.129 mmol, 1.00 equiv) prepared in 4 M HCl (123.47 mg, 3.386 mmol, 3.00 equiv) in dioxane (20.00 mL) was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The residue is purified by reverse phase flash chromatography (C18 silica gel column), eluting with 10-50% ACN in water over 10 minutes and detecting at UV 254 nm to provide 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-63, 200 mg, 41.07%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 432.18.

[1044] Preparation of 2-(2,6-dioxopiperidin-3-yl)-4-[2-(piperazin-1-yl)ethoxy]isoindole-1,3-dione (K-40)

[1045]

[1046] Step 1: 4-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]ethyl Preparation of tert-butyl (1,2-dimethyl)piperazine-1-carboxylate

[1047]

[1048] To a stirred mixture of 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindole-1,3-dione (300.00 mg, 1.094 mmol, 1.00 equiv) and tert-butyl 4-(2-chloroethyl)piperazine-1-carboxylate (299.34 mg, 1.203 mmol, 1.10 equiv) in DMF (5.0 mL) was added KHCO (219.04 mg, 2.188 mmol, 2.00 equiv) and KI (18.16 mg, 0.109 mmol, 0.10 equiv). The resulting mixture was stirred at 60 ° C for 3 h under a nitrogen atmosphere. The mixture was cooled to room temperature, then filtered through a short pad of celite and concentrated in vacuo. The residue was purified by reverse phase flash chromatography to provide tert-butyl 4-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]ethyl)piperazine-1-carboxylate (200 mg, 35.70%) as a yellow solid. LCMS (ESI) m / z [M+H] + = 487.

[1049] Step 2: 2-(2,6-dioxopiperidin-3-yl)-4-[2-(piperazin-1-yl)ethoxy]isoindole-1,3-dione Preparation of (K-40)

[1050]

[1051] A solution of tert-butyl 4-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]ethyl)piperazine-1-carboxylate (200.00 mg, 0.413 mmol, 1.00 equiv) and TFA (1.0 mL) in DCM (3.0 mL) was stirred at 25 °C for 1 h. The resulting mixture was concentrated under reduced pressure to afford crude 2-(2,6-dioxopiperidin-3-yl)-4-[2-(piperazin-1-yl)ethoxy]isoindole-1,3-dione (301 mg) as a white solid, which was used in the next step without further purification. LCMS (ESI) m / z [M+H] + = 387.

[1052] Preparation of 5-((2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-5-yl)oxy)pentanoic acid (K-41) and 5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)pentanoic acid (K-42)

[1053]

[1054] Step 1: 5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)pentanoic acid Preparation of tert-butyl ester

[1055]

[1056] To a solution of 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindole-1,3-dione (1.00 g, 3.647 mmol, 1.00 equiv) and tert-butyl 5-bromopentanoate (0.95 g, 4.011 mmol, 1.10 equiv) in DMF (15.00 mL) was added KI (0.06 g, 0.365 mmol, 0.10 equiv) and KHCO₃ (0.73 g, 7.293 mmol, 2.00 equiv). The resulting mixture was stirred at 60°C overnight. The resulting mixture was diluted with water (100 mL) and extracted with EtOAc (100 mL x 3). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography eluting with 0% to 30% EtOAc in PE to afford tert-butyl 5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)pentanoate (910.0 mg, 52.1%) as a light blue solid. LCMS (ESI) m / z: [M+H] + = 431.

[1057] Step 2: 5-((2-(2,6-dioxopiperidin-3-yl)-1-hydroxy-3-oxoisoindolin-5-yl)oxy)pentane tert-Butyl 5-((2-(2,6-dioxopiperidin-3-yl)-3-hydroxy-1-oxoisoindolin-5-yl)oxy)pentanoic acid Preparation of butyl ester

[1058]

[1059] To a stirred mixture of tert-butyl 5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)pentanoate (910.0 mg, 2.114 mmol, 1.00 equiv) in AcOH (30 mL) was added Zn (2.76 g, 42.281 mmol, 20.00 equiv). The resulting mixture was stirred at 60 °C for 2 hours. The resulting mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography using the following conditions: column, C 18 Silica gel; mobile phase, ACN in water, 10% to 50% gradient in 10 min; detector, UV 254 nm, to provide a mixture of the title compounds as a light yellow solid (1.10 g, crude). LCMS (ESI) m / z [M+H] + =433.

[1060] Step 3: 5-((2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-5-yl)oxy)pentanoic acid and 5- Preparation of ((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)pentanoic acid

[1061]

[1062] To a stirred solution of tert-butyl 5-((2-(2,6-dioxopiperidin-3-yl)-1-hydroxy-3-oxoisoindolin-5-yl)oxy)pentanoate and tert-butyl 5-((2-(2,6-dioxopiperidin-3-yl)-3-hydroxy-1-oxoisoindolin-5-yl)oxy)pentanoate (1.10 g crude) in TFA (30 mL) was added Et3SiH (6 mL). The resulting mixture was stirred at room temperature for 2 hours. The reaction mixture was filtered through a short pad of celite and concentrated in vacuo. The residue was purified by reverse phase flash chromatography using the following conditions: column, C 18 Silica gel; mobile phase, ACN in water, 10% to 50% gradient in 10 min; detector, UV 254 nm, to provide the title compound (820.0 mg, 89.5%). LCMS (ESI) m / z: [M+H] + = 221.10.

[1063] Step 4: Methyl 5-((2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-5-yl)oxy)pentanoate and preparation of methyl 5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)pentanoate

[1064]

[1065] To a stirred solution of 5-((2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-5-yl)oxy)pentanoic acid and 5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)pentanoic acid (820.0 mg, 2.277 mmol) in MeOH (30 mL) was added TMSCHN2 (6 mL) at 0°C. The resulting mixture was stirred at room temperature for 2 hours. The reaction mixture was filtered through a short pad of celite and concentrated in vacuo. The residue was purified by preparative HPLC using the following conditions: column: Xcelect CSH F-pheny OBD column, 19*250 mm, 5 um; mobile phase A: water (0.05% FA), mobile phase B: ACN; flow rate: 25 mL / min; gradient: 24% B to 40% B in 10 min to provide methyl 5-((2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-5-yl)oxy)pentanoate (80.0 mg, 10.1%) and methyl 5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)pentanoate (400.0 mg, 50.6%) as a white solid. LCMS (ESI) m / z: [M+H] + =375.

[1066] Step 5: 5-((2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-5-yl)oxy)pentanoic acid (K- Preparation of 5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)pentanoic acid (K-42)

[1067]

[1068] To a stirred solution of methyl 5-((2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-5-yl)oxy)pentanoate (80.0 mg, 0.213 mmol, 1.00 equiv) in THF (1 mL) was added concentrated HCl (0.5 mL). The resulting mixture was stirred for 1 hour and the mixture was concentrated under vacuum to afford 5-((2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-5-yl)oxy)pentanoic acid (90.0 mg, crude) as a white solid. LCMS (ESI) m / z: [M+H] + =361

[1069] To a stirred solution of methyl 5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)pentanoate (400.0 mg, 1.069 mmol, 1.00 equiv) in THF (3 mL) was added concentrated HCl (1.5 mL). The resulting mixture was stirred for 1 hour and the mixture was concentrated under vacuum to afford 5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)pentanoic acid (500.0 mg, crude) as a white solid. LCMS (ESI) m / z: [M+H] + =361.

[1070] Preparation of 5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oxy)pentanoic acid (K-43)

[1071]

[1072] Step 1: 5-((2-(2,6-dioxopiperidin-3-yl)-3-hydroxy-1-oxoisoindolin-4-yl)oxy)pentane Acid preparation

[1073]

[1074] To a stirred solution of 5-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]pentanoic acid (I-30), (70.0 mg, 0.187 mmol, 1.00 equiv) in AcOH (3 mL) was added Zn (122.3 mg, 1.870 mmol, 10.00 equiv). The resulting mixture was stirred at room temperature for 12 h. The reaction mixture was filtered through a short pad of celite and concentrated in vacuo. This produced crude 5-((2-(2,6-dioxopiperidin-3-yl)-3-hydroxy-1-oxoisoindolin-4-yl)oxy)pentanoic acid (117 mg, crude) as a light yellow solid. LCMS (ESI) m / z [M+H] + =377.

[1075] Step 2: 5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oxy)pentanoic acid (K- Preparation of 43)

[1076]

[1077] To a stirred mixture of 5-((2-(2,6-dioxopiperidin-3-yl)-3-hydroxy-1-oxoisoindolin-4-yl)oxy)pentanoic acid (117.0 mg, 0.311 mmol, 1.00 equiv) and Et3SiH (2 mL) in DCM (4 mL) was added TFA (1 mL) at room temperature. The resulting mixture was stirred for 12 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography using the following conditions: (column: C18 silica gel; mobile phase, ACN in water, 0% to 80% gradient over 40 min; detector, UV 254 nm) to provide 5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oxy)pentanoic acid (17.0 mg, 15.17%) as a white solid. LCMS (ESI) m / z [M+H] + =361.

[1078] Preparation of 5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)amino)pentanoic acid (K-44)

[1079]

[1080] Step 1: Preparation of methyl 2-(bromomethyl)-4-nitrobenzoate

[1081]

[1082] To a stirred solution of methyl 2-methyl-4-nitrobenzoate (2.00 g, 10.247 mmol, 1.00 equiv) in CCl4 (20.00 mL) was added NBS (1.82 g, 0.010 mmol, 1.00 equiv) and BPO (0.21 g, 0.001 mmol, 0.08 equiv) at room temperature. The resulting mixture was stirred at 80 ° C for 3 h. The mixture was cooled to room temperature. The reaction mixture was filtered through a short pad of celite and concentrated in vacuo. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (10: 1) to provide methyl 2-(bromomethyl)-4-nitrobenzoate (2.1 g, 74.77%) as a white solid. LCMS (ESI) m / z: [M+H] + = 274.07.

[1083] Step 2: Preparation of 3-(5-nitro-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[1084]

[1085] To a stirred mixture of methyl 2-(bromomethyl)-4-nitrobenzoate (2.05 g, 7.480 mmol, 1.00 equiv) and aminoglutarimide (1.02 g, 0.009 mmol, 1.20 equiv) in DMF (1.00 mL) was added DIEA (2.90 g, 0.022 mmol, 3.00 equiv) at room temperature. The resulting mixture was stirred at 120°C overnight. The mixture was cooled to room temperature, then filtered through a short pad of celite and concentrated in vacuo. The residue was purified by reverse-phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, ACN in water, gradient from 0% to 100% over 30 min; detector, UV 254 nm, to afford 3-(5-nitro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (940 mg, 34.76%) as a black solid. LCMS (ESI) m / z: [M+H]+ = 289.25.

[1086] Step 3: Preparation of 3-(5-amino-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[1087]

[1088] To a stirred solution of 3-(5-nitro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (380.00 mg, 1.314 mmol, 1.00 equiv) in AcOH (10.00 mL) was added Zn (859.31 mg, 13.138 mmol, 10 equiv) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was filtered and the filter cake was washed with H2O (3x3 mL). The filtrate was concentrated under reduced pressure to afford 3-(5-amino-1-oxoisoindolin-2-yl)piperidine-2,6-dione (320 mg, 56.37%) as a black solid, which was used in the next step without further purification. LCMS (ESI) m / z: [M+H]+ = 259.27.

[1089] Step 4: Preparation of methyl 5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)amino)pentanoate preparation

[1090]

[1091] To a stirred mixture of 3-(5-amino-1-oxoisoindolin-2-yl)piperidine-2,6-dione (600.00 mg, 2.314 mmol, 1.00 equiv) and methyl 5-oxopentanoate (451.77 mg, 3.471 mmol, 1.50 equiv) in MeOH (20.00 mL) was added NaBHCN (436.29 mg, 6.943 mmol, 3.00 equiv) and AcOH (0.60 mL, 9.981 mmol, 4.52 equiv) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The reaction mixture was filtered through a short pad of celite and concentrated in vacuo. The residue was purified by reverse phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, ACN in water, gradient from 0% to 100% over 30 min; detector, UV 254 nm to afford methyl 5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)amino)pentanoate (630 mg, 67.80%) as a gray solid. LCMS (ESI) m / z: [M+H]+ = 373.41.

[1092] Step 5: 5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)amino)pentanoic acid (K- Preparation of 44)

[1093]

[1094] A solution of methyl 5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)amino)pentanoate (600 mg, 1.607 mmol, 1.00 equiv) in 2N HCl (15.00 mL, 30.000 mmol, 18.67 equiv) was stirred at room temperature overnight. The reaction mixture was filtered through a short pad of celite and concentrated in vacuo. The residue was purified by reverse phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, ACN in water, 0% to 100% gradient over 30 min; detector, UV 254 nm to afford the title compound (233 mg, 38.74%) as a grey solid. LCMS (ESI) m / z: [M+H]+ = 359.38.

[1095] Preparation of 4-[3-(3-aminopropyl)azetidin-1-yl]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (K-45)

[1096]

[1097] Step 1: Preparation of 3-(azetidin-3-yl)propan-1-ol

[1098]

[1099] To a solution of tert-butyl 3-(3-hydroxypropyl)azetidine-1-carboxylate (400.00 mg, 1.858 mmol, 1.00 equivalent) in DCM (4.00 mL) was added TFA (4.00 mL). The mixture was stirred at room temperature for 2 h. The reaction mixture was concentrated to provide 3-(azetidine-3-yl)propane-1-ol (200 mg, crude) as a yellow oil.

[1100] Step 2: 2-(2,6-dioxopiperidin-3-yl)-4-[3-(3-hydroxypropyl)azetidin-1-yl]isoindole Preparation of Indole-1,3-Dione

[1101]

[1102] To a stirred mixture of 3-(azetidin-3-yl)propan-1-ol (200.00 mg, 1.736 mmol, 1.00 equiv) and 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindole-1,3-dione (479.65 mg, 1.736 mmol, 1.00 equiv) in DMSO (1 mL) at room temperature under a nitrogen atmosphere was added DIEA (2244.27 mg, 17.365 mmol, 10.00 equiv). The reaction mixture was stirred at 80°C for 16 h. The reaction solution was purified by reverse-phase flash column chromatography using the following conditions: column: C18 spherical gel; mobile phase A: water, mobile phase B: MeOH; flow rate: 40 mL / min; gradient: 0% B to 100% B over 20 min; 254 / 220 nm. This resulted in 2-(2,6-dioxopiperidin-3-yl)-4-[3-(3-hydroxypropyl)azetidin-1-yl]isoindole-1,3-dione (300 mg, 46.52%) as a yellow oil. LCMS (ESI) m / z [M+H] + =372.

[1103] Step 3: 3-[1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]azetidine Preparation of [alkyl-3-yl] propanal

[1104]

[1105] To a stirred mixture of 2-(2,6-dioxopiperidin-3-yl)-4-[3-(3-hydroxypropyl)azetidin-1-yl]isoindole-1,3-dione (505.0 mg, 1.360 mmol, 1.00 equiv) in DCM (10.00 mL) was added Dess-Martin periodinane at room temperature under a nitrogen atmosphere. The mixture was stirred at room temperature overnight. The mixture was purified by preparative TLC (PE / EtOAc 1:1) to afford 3-[1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]azetidin-3-yl]propanal (322 mg, 64.2%) as a yellow solid. LCMS (ESI) m / z [M+H] + =370.

[1106] Step 4: 4-[3-(3-[[(2,4-dimethoxyphenyl)methyl]amino]propyl)azetidin-1-yl]-2- Preparation of (2,6-dioxopiperidin-3-yl)isoindole-1,3-dione

[1107]

[1108] To a stirred solution of 3-[1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl]azetidin-3-yl]propanal (200.00 mg, 0.541 mmol, 1.00 equiv) and 1-(2,4-dimethoxyphenyl)methanamine (181.07 mg, 1.082 mmol, 2.00 equiv) in MeOH (10 mL) was added AcOH (97.55 mg, 1.623 mmol, 3.00 equiv) and NaBH3CN (102.08 mg, 1.623 mmol, 3.00 equiv) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 16 h. The reaction was quenched with water / ice at 0 ° C. The resulting mixture was extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (2 x 20 mL) and then dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by flash chromatography under the following conditions: column: C18 spherical gel; mobile phase A: water, mobile phase B: ACN; flow rate: 40 mL / min; gradient: 0% B to 100% B in 20 min; 254 / 220 nm. This produced 4-[3-(3-[[(2,4-dimethoxyphenyl)methyl]amino]propyl)azetidin-1-yl]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (150 mg, 53.22%) as a yellow solid. LCMS (ESI) m / z [M+H] + =521.

[1109] Step 5: N-[(2,4-dimethoxyphenyl)methyl]-N-(3-[1-[2-(2,6-dioxopiperidin-3-yl)-1, Preparation of tert-butyl 3-(3-dioxoisoindol-4-yl)azetidin-3-yl)propyl)carbamate

[1110]

[1111] To a stirred solution of 4-[3-(3-[[(2,4-dimethoxyphenyl)methyl]amino]propyl)azetidin-1-yl]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (70.00 mg, 0.134 mmol, 1.00 equiv) and DIEA (52.14 mg, 0.403 mmol, 3.00 equiv) in DCM (4.00 mL) was added BoC2O (58.69 mg, 0.269 mmol, 2.00 equiv) at 0° C. under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 3 h and then concentrated under vacuum. The crude product was purified by flash chromatography using the following conditions: Column: C18 spherical gel; Mobile phase A: Water, Mobile phase B: ACN; Flow rate: 40 mL / min; Gradient: 0% B to 100% B over 20 min; 254 / 220 nm. This yielded tert-butyl N-[(2,4-dimethoxyphenyl)methyl]-N-(3-[1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl]azetidin-3-yl]propyl)carbamate (48.0 mg, 57.51%) as a yellow solid. LCMS (ESI) m / z [M+H] + =621.

[1112] Step 6: 4-[3-(3-aminopropyl)azetidin-1-yl]-2-(2,6-dioxopiperidin-3-yl)isoindole Preparation of Indole-1,3-dione (K-45)

[1113]

[1114] A solution of tert-butyl N-[(2,4-dimethoxyphenyl)methyl]-N-(3-[1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl]azetidin-3-yl]propyl)carbamate (48.00 mg, 0.077 mmol, 1.00 equiv) in neat trifluoroacetic acid (1.00 mL) was stirred at room temperature under a nitrogen atmosphere for 6 h. The resulting mixture was concentrated under vacuum. The crude product was purified by flash chromatography using the following conditions: Column: C18 spherical gel; Mobile phase A: Water, Mobile phase B: ACN; Flow rate: 40 mL / min; Gradient: 0% B to 100% B over 20 min; 254 / 220 nm. This resulted in 4-[3-(3-aminopropyl)azetidin-1-yl]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3- (21.0 mg, 73.31%) as a yellow solid. LCMS (ESI) m / z [M+H] + =371.

[1115] Preparation of (2S,4R)-1-((S)-3,3-dimethyl-2-(2-oxoacetylamino)butyryl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (K-46)

[1116]

[1117] 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 (50.00 mg, 0.116 mmol, 1.00 equiv) and glyoxylic acid (8.60 mg, 0.116 mmol, 1.00 equiv) in DMF (1.00 mL) was added HATU (66.23 mg, 0.174 mmol, 1.50 equiv) and DIEA (45.03 mg, 0.348 mmol, 3.00 equiv) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The residue was purified by reverse phase flash chromatography (eluting with 0-100% acetonitrile / water containing 0.1% formic acid) to provide the title compound (65 mg) as a yellow oil. LCMS (ESI) m / z: [M+H] + =487.

[1118] Preparation of (2S,4R)-1-((S)-3,3-dimethyl-2-(11-oxoundecanoylamino)butyryl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (K-47)

[1119]

[1120] Step 1: Preparation of 11-oxoundecanoic acid

[1121]

[1122] At -78 ℃ under nitrogen atmosphere to (COCl) 2 (2.51 g, 19.773 mmol, 4.00 equivalents) in CH 2 Cl 2 (10mL) in stirred solution was dropwise added DMSO (1.16 g, 14.830 mmol, 3.00 equivalents). Then add 11-hydroxy undecanoic acid (1.00 g, 4.943 mmol, 1.00 equivalents) in CH 2 Cl 2 (10mL) in solution, and the mixture was stirred at -60 ℃ for 30 min under nitrogen atmosphere. Then add Et 3 N (2.50 g, 24.717 mmol, 5.00 equivalents) at -60 ℃ and the mixture was warmed to room temperature and stirred for another 1.5 h. The mixture obtained was diluted with water (50 mL), then extracted with CH 2 Cl 2 (50mL x 3). The combined organic layer was washed with salt water (30 mL), then dried over anhydrous Na 2 SO 4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was used directly in the next step without further purification. This gave the title compound as a yellow oil (950 mg, crude). LCMS (ESI) m / z: [M+H] + =200.

[1123] Step 2: (2S,4R)-1-((S)-3,3-dimethyl-2-(11-oxoundecanoylamino)butanoyl)-4-hydroxy Preparation of 1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (K-47)

[1124]

[1125] To a stirred solution of 11-oxoundecanoic acid (100.00 mg, 0.499 mmol, 1.00 equiv) and (2S,4R)-1-[(2S)-2-amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (266.38 mg, 1.20 equiv) in DMF (5 mL) was added HATU (284.78 mg, 0.748 mmol, 1.50 equiv) and DIEA (193.60 mg, 1.497 mmol, 3.00 equiv) portionwise at room temperature. The resulting mixture was stirred for 6 h and then concentrated. The residue was purified by reverse phase flash chromatography to provide the title compound (90 mg, 28.74%) as a white solid. LCMS (ESI) m / z: [M+H] + =257.

[1126] Preparation of (2S,4R)-1-((S)-2-acrylamido-3,3-dimethylbutyryl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (K-48)

[1127]

[1128] To a stirred room temperature mixture 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 (500.00 mg, 1.161 mmol, 1.00 equiv) and acrylic acid (83.68 mg, 1.161 mmol, 1.00 equiv) in DMF (4.00 mL) was added HATU (529.85 mg, 1.393 mmol, 1.20 equiv) and DIEA (450.25 mg, 3.483 mmol, 3.00 equiv). The resulting mixture was stirred for 2 h and then concentrated. The residue was purified by reverse phase flash chromatography (eluting with 10-50% acetonitrile in water over 30 min) to provide the title compound as a yellow oil (495 mg, 87.96%). LCMS (ESI) m / z [M+H] + =484.21.

[1129] Preparation of (2S,4R)-4-hydroxy-1-((R)-3-methyl-2-(3-(2-oxoethoxy)isoxazol-5-yl)butyryl)-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (K-49)

[1130]

[1131] Step 1: Preparation of 2-(3-bromoisoxazol-5-yl)ethan-1-ol

[1132]

[1133] By 3-butyn-1-ol (552.89 g, 7888.26 mmol, 4 equivalents) and KHCO (592.30 g, 5916.197 mmol, 3 equivalents) in EtOAc (2600 mL) and H o (260 mL) solution is stirred at room temperature. Last 60min dropwise addition 1,1-dibromoformaldehyde oxime (1-bromo-N-hydroxymethanecarbonimidoyl bromide) (400.00 g in EA (840 mL), 1972.066 mmol, 1.00 equivalents). The resulting mixture is stirred at room temperature overnight. The reaction mixture is washed with water (500 mL x 2) and the organic layer is passed through anhydrous Na sO dried. After filtration, the filtrate is concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (30:1) to provide the title compound as an off-white solid (338.2 g, 88.98%). LCMS (ESI) m / z [M+H] + =192.

[1134] Step 2: Preparation of 2-(3-bromoisoxazol-5-yl)acetic acid

[1135]

[1136] A solution of 2-(3-bromoisoxazol-5-yl)ethane-1-ol (360.00 g) in acetone (3600 mL) was stirred at 0 ° C under a nitrogen atmosphere. Jones reagent (1760.00 mL) was added dropwise to the above mixture at 0 ° C for 1 h. The resulting mixture was stirred at room temperature overnight. The reaction was quenched by adding ice water, and the resulting mixture was extracted with EtOAc (1000 mL x 3). The combined organic layers were washed with water (500 mL x 2) and the organic layer was dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to provide the title compound (348.6 g, crude) as a green solid, which was used directly without further purification. (LCMS (ESI) m / z [M+H] + =206.

[1137] Step 3: Preparation of ethyl 2-(3-bromoisoxazol-5-yl)acetate

[1138]

[1139] A solution of 2-(3-bromoisoxazol-5-yl)acetic acid (397.6 g, 1930.144 mmol, 1.00 equiv) and H2SO4 (18.92 g, 193.014 mmol, 0.1 equiv) in EtOH (2000 mL) was stirred at 70°C for 2 h. The reaction mixture was concentrated under reduced pressure. The resulting mixture was diluted with EtOAc (3000 mL), washed with water (500 mL x 2), and the organic layer was dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (35:1) to provide the title compound (355 g, 78.61%) as a colorless oil. (LCMS(ESI) m / z [M+H] + =234.

[1140] Step 4: Preparation of ethyl 2-(3-bromoisoxazol-5-yl)-3-methylbutanoate

[1141]

[1142] To a stirred solution of t-BuOK (244.51 g, 2179.031 mmol, 1.5 equivalents) and ethyl 2-(3-bromoisoxazole-5-yl)acetate (340.00 g, 1452.687 mmol, 1.00 equivalents) in THF (2000 mL) was added 2-iodopropane (321.03 g, 1888.493 mmol, 1.3 equivalents) dropwise at 0 ° C under a nitrogen atmosphere. The resulting mixture was stirred at room temperature overnight and then ice-cold water was added. The mixture was extracted with EtOAc (1000 mL x 2). The combined organic layer was washed with water (500 mL x 1) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / THF (10:1) to afford the title compound (284.1 g, 70.82%) as a colorless oil. (LCMS (ESI) m / z [M+H] + =276.

[1143] Step 5: Preparation of 2-(3-methoxyisoxazol-5-yl)-3-methylbutanoic acid

[1144]

[1145] To a stirred solution of ethyl 2-(3-bromoisoxazol-5-yl)-3-methylbutanoate (90.00 g, 325.933 mmol, 1.00 equiv) in MeOH (270 mL) was added a solution of KOH (274.30 g, 4888.995 mmol, 15.00 equiv) in MeOH (210 mL) at 0 ° C. The reaction mixture was stirred at 80 ° C overnight. The resulting solution was acidified to pH 4 with 1M HCl solution (aqueous solution) and concentrated under reduced pressure. The resulting mixture was diluted with EtOAc (1800 mL) and filtered. The filter cake was washed with EtOAc (100 mL x 3). The filtrate was concentrated under reduced pressure to provide the title compound (62.9 g, 96.88%) as a yellow oil, which was used directly without further purification. LCMS (ESI) m / z: [M+H] + =200.

[1146] Step 6: Preparation of 2-(3-hydroxyisoxazol-5-yl)-3-methylbutanoic acid

[1147]

[1148] To a stirred solution of 2-(3-methoxyisoxazol-5-yl)-3-methylbutanoic acid (62.90 g, 315.754 mmol, 1.00 equiv) in HOAc (450.00 mL) was added 48% HBr (450.00 mL) at room temperature. The resulting mixture was stirred at 60 °C for 16 h and then concentrated under reduced pressure to produce a residue. The residue was purified by flash C18-flash chromatography eluting with 0-100% MeCN in water / 0.05% formic acid to provide the title compound (43.3 g, 74.05%) as a white solid. LCMS (ESI) m / z: [M+H] + = 186.

[1149] Step 7: Preparation of methyl 2-(3-hydroxyisoxazol-5-yl)-3-methylbutanoate

[1150]

[1151] To a stirred solution of 2-(3-hydroxyisoxazole-5-yl)-3-methylbutanoic acid (20 g, 108.004 mmol, 1.00 equivalent) in MeOH (72 mL) was added SOCl2 (35.26 mL, 486.059 mmol, 4.50 equivalent) at 0 ° C. The resulting mixture was stirred at room temperature for 16 h, then concentrated under reduced pressure to produce a residue. The residue was diluted with water (30 mL) and extracted with EtOAc (50 mL x 3). The combined organic layer was washed with saturated brine (30 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography, eluted with 0-100% THF in petroleum ether to provide the title compound (15.1 g, 70.18%) as a pale solid. 1 H NMR (400 MHz, DMSO- d 6) δ 11.24 (s, 1H), 5.95 (s, 1H), 3.71 - 3.58 (m, 4H), 2.32 - 2.20 (m,1H), 0.88 (dd, J = 34.2, 6.7 Hz, 6H). LCMS (ESI) m / z: [M+H] + = 200.

[1152] Step 8: Preparation of methyl 2-(3-(2,2-diethoxyethoxy)isoxazol-5-yl)-3-methylbutanoate

[1153]

[1154] To a stirred solution of methyl 2-(3-hydroxyisoxazol-5-yl)-3-methylbutanoate (7 g, 35.140 mmol, 1.00 eq) and 2-bromo-1,1-diethoxyethane (7.62 g, 38.654 mmol, 1.1 eq) in DMF (70 mL) was added K2CO3 (9.71 g, 70.280 mmol, 2 eq). The resulting mixture was stirred at 80 ° C overnight. The resulting mixture was diluted with EtOAc (300 mL). The organic layer was washed with water (300 mL), brine (300 mL), then dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by preparative HPLC to provide the title compound (5.2 g, 46.92%) as a brown solid. LCMS (ESI) m / z: [M+H] + = 316.

[1155] Step 9: 2-(3-(2,2-diethoxyethoxy)isoxazol-5-yl)-3-methylbutanoic acid

[1156]

[1157] A solution of methyl 2-(3-(2,2-diethoxyethoxy)isoxazol-5-yl)-3-methylbutanoate (5.2 g, 16.489 mmol, 1.00 equiv) and LiOH (1.97 g, 82.445 mmol, 5 equiv) in MeOH (15 mL) and H2O (45 mL) was prepared, and the resulting mixture was stirred at room temperature for 2 h. The mixture was acidified to pH 5 with concentrated HCl. The resulting mixture was extracted with EtOAc (300 mL x 3). The combined organic layers were washed with brine (100 mL x 1) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure and the filtrate was concentrated under vacuum. The crude product was used directly in the next step without further purification. LCMS (ESI) m / z: [M+H] + = 302.

[1158] Step 10: (2S,4R)-1-((R)-2-(3-(2,2-diethoxyethoxy)isoxazol-5-yl)-3-methylbutane Preparation of 4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[1159]

[1160] To a stirred solution of 2-(3-(2,2-diethoxyethoxy)isoxazol-5-yl)-3-methylbutanoic acid (4.95 g, 16.427 mmol, 1.00 equiv) and (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (5.44 g, 16.427 mmol, 1 equiv) in DMF (50 mL) was added HATU (6.87 g, 18.070 mmol, 1.1 equiv) and DIEA (6.37 g, 49.281 mmol, 3 equiv). The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was diluted with EtOAc (300 mL). The organic layer was washed with water (300 mL), brine (300 mL), then dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography eluting with PE / EA (1:1) to provide a crude product. The crude product (6.2 g) was purified by chiral-HPLC using a CHIRALART Amylose-SA column eluted with 40% methanol in carbon dioxide to provide the title compound (2.8 g, 27.73%) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ 8.99 (d, J = 3.4 Hz, 1H), 8.43(d, J= 7.7 Hz, 1H), 7.49 - 7.41 (m, 2H), 7.41 - 7.31 (m, 2H), 6.14 (s, 1H),5.10 (d, J = 3.6 Hz, 1H), 4.97 - 4.87 (m, 1H), 4.81 (t, J = 5.2 Hz, 1H), 4.37(t, J = 7.9 Hz, 1H), 4.32 - 4.23 (m, 1H), 4.09 (d, J = 5.3 Hz, 2H), 3.73 -3.49 (m, 6H), 3.45 (d, J = 10.8 Hz, 1H), 2.46 (d, J = 2.1 Hz, 3H), 2.31 -2.15 (m, 1H), 2.03 (ddd, J = 11.9, 8.1, 3.0 Hz, 1H), 1.78 (ddd, J = 12.8,8.0, 4.7 Hz, 1H), 1.41 (dd, J = 29.6, 7.0 Hz, 3H), 1.13 (t, J = 7.0 Hz, 6H),0.96 (t, J = 6.4 Hz, 3H), 0.81 (dd, J = 14.4, 6.7 Hz, 3H)。LCMS (ESI) m / z: [M+H] + = 615.35。

[1161] Step 11: (2S,4R)-4-hydroxy-1-((R)-3-methyl-2-(3-(2-oxoethoxy)isoxazol-5-yl) Preparation of (4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (K-49)

[1162]

[1163] To a stirred solution of H2SO4 (1M) (6.00 mL) and THF (6.00 mL) was added (2S,4R)-1-((R)-2-(3-(2,2-diethoxyethoxy)isoxazol-5-yl)-3-methylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (300.00 mg, 0.499 mmol, 1.00 equiv) at room temperature. The resulting mixture was stirred at 50 °C for 8 h. The reaction was quenched by the addition of ice water, and the mixture was basified to pH 7 with saturated NaHCO3(aq). The resulting mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (2 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to provide (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]-1-[(2R)-3-methyl-2-[3-(2-oxoethoxy)-1,2-oxazol-5-yl]butanoyl]pyrrolidine-2-carboxamide (256 mg, 97.3%) as a white solid, which was used directly without further purification. LCMS (ESI) m / z: [M+H] + =541.

[1164] Preparation of 4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)oxy)ethyl)benzoic acid (I-64)

[1165]

[1166] Step 1: Preparation of 4-(2-((3-amino-6-chloropyridazin-4-yl)oxy)ethyl)-N-methylbenzamide

[1167]

[1168] To a solution of 4-(2-hydroxyethyl)-N-methylbenzamide (1.00 g, 5.580 mmol, 1.00 equiv) in THF (30.00 mL) was added portionwise NaH (333.33 mg, 8.370 mmol, 1.50 equiv, 60% in oil) at 0°C under an atmosphere of dry nitrogen. The resulting mixture was stirred at 0°C for 30 minutes and 4-bromo-6-chloropyridazin-3-amine (1.40 g, 6.696 mmol, 1.20 equiv) was added at room temperature. The mixture was heated to 60°C and stirred for 6 hours. The mixture was poured into a saturated aqueous NH4Cl solution (20 mL) and extracted with DCM (3 x 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated, and the residue was purified by silica gel column chromatography eluting with DCM to 20:1 DCM / MeOH to provide 4-(2-((3-amino-6-chloropyridazin-4-yl)oxy)ethyl)-N-methylbenzamide (546 mg, 28.39%) as a light yellow solid. LCMS (ESI) m / z: [M+H] + = 307.

[1169] Step 2: 4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)oxy)ethyl)-N-methylbenzoyl Preparation of amines

[1170]

[1171] A mixture of 4-(2-((3-amino-6-chloropyridazin-4-yl)oxy)ethyl)-N-methylbenzamide (546.00 mg, 1.780 mmol, 1.00 equiv) and 2-hydroxyphenylboronic acid (368.26 mg, 2.670 mmol, 1.50 equiv) in 4:1 dioxane / water (10.00 mL) was sparged with nitrogen for 5 minutes. Pd(dppf)Cl2 (130.24 mg, 0.178 mmol, 0.1 equiv) and K2CO3 (45.05 mg, 0.326 mmol, 2.0 equiv) were added. The resulting mixture was stirred at 80°C for 4 h. The mixture was cooled to room temperature and quenched with water. The resulting mixture was extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (20 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure and purified to afford 4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)oxy)ethyl)-N-methylbenzamide (429 mg, 62.83%) as a brown solid. LCMS (ESI) m / z [M+H] + = 365.

[1172] Step 3: Reaction of 4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)oxy)ethyl)benzoic acid (I-64) preparation

[1173]

[1174] A mixture of 4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)oxy)ethyl)-N-methylbenzamide (429.00 mg, 1.179 mmol, 1.00 equiv) in concentrated HCl (10.00 mL, 12 M) was stirred at 90 °C for 8 h. The mixture was cooled to room temperature, diluted with water (5 mL), neutralized with NaHCO3 to pH 3, and filtered. The filter cake was further purified by reverse phase preparative HPLC to afford 4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)oxy)ethyl)benzoic acid (I-64, 389 mg, 90.91%) as a white solid. LCMS (ESI) m / z [M+H] + = 352.20.

[1175] Preparation of (4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)oxy)ethyl)phenyl)(3-(piperidin-4-yl)azetidin-1-yl)methanone (K-50)

[1176]

[1177] Step 1: 4-[1-[4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]oxy]ethyl)benzoyl Preparation of tert-butyl]azetidin-3-yl]piperidine-1-carboxylate

[1178]

[1179] To a stirred solution of 4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]oxy]ethyl)benzoic acid (50.0 mg, 0.142 mmol, 1.00 equiv) and tert-butyl 4-(azetidin-3-yl)piperidine-1-carboxylate (34.2 mg, 0.142 mmol, 1.00 equiv) in DMF (2.0 mL) was added HATU (64.9 mg, 0.171 mmol, 1.20 equiv) and DIEA (36.7 mg, 0.285 mmol, 2.00 equiv) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The mixture was purified by reverse phase flash chromatography using the following conditions (column, C18 silica gel; mobile phase, MeCN in water (0.1% FA), 0-100% gradient over 30 min; detector, UV 254 / 220 nm). This gave the title compound as an off-white solid (55 mg, 57.37%). LCMS (ESI) m / z: [M+H]+ = 574.

[1180] Step 2: 2-[6-amino-5-(2-[4-[3-(piperidin-4-yl)azetidine-1-carbonyl]phenyl]ethoxy) Preparation of [3-pyridazinyl]phenol (K-50)

[1181]

[1182] To a stirred solution of tert-butyl 4-[1-[4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]oxy]ethyl)benzoyl]azetidin-3-yl]piperidine-1-carboxylate (55.0 mg, 0.096 mmol, 1.00 equiv) in DCM (2.0 mL) was added TFA (1.0 mL) at room temperature. The resulting solution was stirred for 2 h and the resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography using the following conditions (column, C18 silica gel; mobile phase, MeCN in water (0.1% FA), 0-100% gradient in 30 min; detector, UV 254 / 220 nm). This produced the title compound (20 mg, 44.05%) as a pink solid. LCMS (ESI) m / z: [M+H] + = 474.

[1183] In a similar manner as described in the preparation of K-50, the following intermediates in Table A13 were prepared.

[1184]

[1185]

[1186]

[1187] Preparation of 2-(6-amino-5-[2-[4-(aminomethyl)phenyl]ethoxy]pyridazin-3-yl)phenol FA (I-65)

[1188]

[1189] Step 1: Preparation of 2-(4-(aminomethyl)phenyl)ethan-1-ol

[1190]

[1191] To a stirred mixture of 4-(2-hydroxyethyl)benzonitrile (2.00 g, 13.589 mmol, 1.00 equiv) in MeOH (50.0 mL) was added concentrated HCl (1.0 mL) and wet 10% Pd / C (1.00 g) at room temperature. The reaction vessel was purged three times with hydrogen and the resulting mixture was stirred at room temperature for 16 h under 5 atmospheres of hydrogen. The reaction vessel was sparged with nitrogen and the catalyst was removed by Celite® filtration. The filter cake was washed with MeOH and the filtrate was concentrated in vacuo to provide 2-(4-(aminomethyl)phenyl)ethane-1-ol (2.08 g, quantitative) as a light brown solid, which was used in the next step without further purification. LCMS (ESI) m / z [M+H] + = 152.

[1192] Step 2: Preparation of tert-butyl 4-(2-hydroxyethyl)benzyl)carbamate

[1193]

[1194] To a stirred mixture of 2-(4-(aminomethyl)phenyl)ethane-1-ol (2.00 g, 1.00 equivalent, 13.23 mmol) in DCM (30.00 mL) was added portionwise Boc O (4.33 g, 1.50 equivalent, 19.840 mmol) and TEA (2.677 g, 2.00 equivalent, 26.454 mmol) at 0 ° C. The reaction mixture was stirred at room temperature for 4 h and then quenched with water (10 mL). The resulting mixture was extracted with DCM (3 x 50 mL) and the combined organic layers were washed with brine (10 mL), dried over anhydrous Na SO dried, filtered, and concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography (C18 column; mobile phase, ACN in water, gradient 0% to 80% over 30 min; detector, UV 254 nm) to provide tert-butyl 4-(2-hydroxyethyl)benzyl)carbamate (2.14 g, 64.46%) as a white solid. LCMS (ESI) m / z [M+H] + = 252.

[1195] Step 3: tert-N-[(4-[2-[(3-amino-6-chloropyridazin-4-yl)oxy]ethyl]phenyl)methyl]carbamic acid Preparation of butyl ester

[1196]

[1197] To a stirred mixture of tert-butyl 4-(2-hydroxyethyl)benzyl)carbamate (2.14 g, 8.53 mmol, 1.00 equiv) in DMF (20 mL) at 0 °C was added portionwise t-BuOK (1.43 g, 12.79 mmol, 1.50 equiv). The reaction mixture was stirred at 0 ° C for 45 min. A solution of 4-bromo-6-chloropyridazin-3-amine (1.78 g, 8.53 mmol, 1.00 equiv) in anhydrous DMF (10 mL) was added dropwise at room temperature. The reaction mixture was heated at 60 ° C for 4 h and then quenched with water (50 mL). The resulting mixture was extracted with DCM (3 x 50 mL) and the combined organic layers were washed with brine (100 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography to provide tert-butyl N-[(4-[2-[(3-amino-6-chloropyridazin-4-yl)oxy]ethyl]phenyl)methyl]carbamate (1.65 g, 51.17%) as a brown solid. LCMS (ESI) m / z [M+H] + = 379.

[1198] Step 4: N-[[4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]oxy]ethyl)phenyl]methyl] Preparation of tert-butyl carbamate

[1199]

[1200] To a stirred mixture of tert-butyl N-[(4-[2-[(3-amino-6-chloropyridazin-4-yl)oxy]ethyl]phenyl)methyl]carbamate (1.00 g, 2.640 mmol, 1.00 equiv) and 2-hydroxyphenylboronic acid (545 mg, 3.960 mmol, 1.50 equiv) in dioxane (12.0 mL) and water (3.0 mL) was added KCO (0.729 g, 5.280 mmol, 2.0 equiv) and Pd(PPh) (305.184 mg, 0.264 mmol, 0.10 equiv) at room temperature under an atmosphere of dry nitrogen. The resulting mixture was stirred at 80° C. under an atmosphere of dry nitrogen for 4 h. The mixture was cooled to room temperature and diluted with water (10 mL). The resulting mixture was extracted with EtOAc (3 x 50 mL). The combined organic layers were dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to provide tert-butyl N-[[4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]oxy]ethyl)phenyl]methyl]carbamate (765 mg, 66.46%) as a white solid. LCMS (ESI) m / z [M+H] + = 437.

[1201] Step 5: 2-(6-amino-5-[2-[4-(aminomethyl)phenyl]ethoxy]pyridazin-3-yl)phenol FA (I- Preparation of 65)

[1202]

[1203] To a stirred mixture of tert-butyl N-[[4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]oxy]ethyl)phenyl]methyl]carbamate (200.00 mg, 0.459 mmol, 1.00 equiv) in DCM (10 mL) was added TFA (2 mL) dropwise at room temperature under an atmosphere of dry nitrogen. The resulting mixture was stirred at room temperature for 1 h and then concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography to provide intermediate I-65 (98.00 mg, 55.9%) as a white solid. LCMS (ESI) m / z [M+H] + = 337.25.

[1204] In a similar manner as described in the preparation of 1-65, the following intermediates in Table A14 were prepared.

[1205]

[1206] Preparation of N-(4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)oxy)ethyl)benzyl)-2-(2,6-diazaspiro[3.3]heptane-2-yl)acetamide (K-65)

[1207]

[1208] Step 1: Preparation of tert-butyl 6-(2-ethoxy-2-oxoethyl)-2,6-diazaspiro[3.3]heptane-2-carboxylate Preparation

[1209]

[1210] To a stirred solution of tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate hemioxalate (200.00 mg, 0.694 mmol, 1.00 equiv) and ethyl bromoacetate (139.02 mg, 0.832 mmol, 1.20 equiv) in DMF (4.00 mL) was added KCO (239.69 mg, 1.734 mmol, 2.5 equiv) at room temperature. The resulting mixture was stirred at 50 ° C for 2 h. The residue was purified by reverse phase flash chromatography to provide the title compound (280 mg, crude) as a white solid. LCMS (ESI) m / z: [M + H] + =285.

[1211] Step 2: Preparation of 2-(6-(tert-butoxycarbonyl)-2,6-diazaspiro[3.3]heptane-2-yl)acetic acid

[1212]

[1213] A solution of tert-butyl 6-(2-ethoxy-2-oxoethyl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (200.00 mg, 0.703 mmol, 1.00 equiv) and LiOH (67.37 mg, 2.813 mmol, 4.00 equiv) in MeOH (2.00 mL) and H2O (0.50 mL) was stirred at 60 °C for 2 h. The resulting mixture was concentrated under reduced pressure to afford the title compound (250 mg, crude) as a white solid. LCMS (ESI) m / z: [M+H] + =257.

[1214] Step 3: 6-(2-((4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)oxy)ethyl)benzyl)amino Preparation of tert-butyl (2-(2-methyl)-2-oxoethyl)-2,6-diazaspiro[3.3]heptane-2-carboxylate

[1215]

[1216] To a stirred solution of 2-(6-amino-5-(4-(aminomethyl)phenethoxy)pyridazin-3-yl)phenol (150.00 mg, 0.585 mmol, 1.00 equiv) and 2-(6-(tert-butoxycarbonyl)-2,6-diazaspiro[3.3]heptan-2-yl)acetic acid (196.87 mg, 0.585 mmol, 1.00 equiv) in DMF (4.00 mL) was added HATU (222.53 mg, 0.585 mmol, 1 equiv) and DIEA (226.92 mg, 1.756 mmol, 3 equiv) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The residue was purified by reverse phase flash chromatography to afford the title compound (37 mg, 11.00%) as a white solid. LCMS (ESI) m / z: [M+H] + =575.

[1217] Step 4: N-[[4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]oxy]ethyl)phenyl]methane Preparation of 2-[2,6-diazaspiro[3.3]heptan-2-yl]acetamide (K-65)

[1218]

[1219] To a stirred solution of tert-butyl 6-(2-((4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)oxy)ethyl)benzyl)amino)-2-oxoethyl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (37.00 mg, 0.064 mmol, 1.00 equiv) in DCM (1.20 mL) was added TFA (0.40 mL) dropwise at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, MeCN in water (0.1% FA), 10% to 50% gradient over 10 min; detector, UV 254 nm. To afford the title compound (26.80 mg, 87.86%) as a white solid. LCMS (ESI) m / z: [M+H] + =475.

[1220] In a similar manner as described in the preparation of K-65, the following intermediates in Table A15 were prepared:

[1221]

[1222]

[1223]

[1224] Preparation of 4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl](methyl)amino]ethyl)benzoic acid (I-66)

[1225]

[1226] Step 1: Preparation of tert-butyl N-[2-[4-(dimethylcarbamoyl)phenyl]ethyl]carbamate

[1227]

[1228] To the stirred solution of 4-[2-[(tert-butoxycarbonyl)amino]ethyl]benzoic acid (5.0 g, 18.8 mmol, 1.0 equivalent) in DMF (50.0 mL), add DIEA (7.3 g, 56.5 mmol, 3.0 equivalents) and HATU (1.3 equivalents).The mixture obtained is stirred at room temperature for 30 min, then dimethylamine (1.02 g, 22.6 mmol, 1.2 equivalents) is added. The mixture obtained is stirred at room temperature for other 2 h, then diluted with water, and extracted with EtOAc (3 x 15 mL). The organic layer merged is washed with salt water (15 mL), through anhydrous Na sO dry, filter, and under reduced pressure concentrate to provide N-[2-[4-(dimethylcarbamoyl)phenyl]ethyl]t-butyl carbamate (5.9 g) as a yellow oil. The crude product is used in the next step without being further purified. LCMS (ESI) m / z: [M+H] + = 237.1.

[1229] Step 2: Preparation of tert-butyl N-[2-[4-(dimethylcarbamoyl)phenyl]ethyl]-N-methylcarbamate

[1230]

[1231] In a 250 mL round-bottom flask, tert-butyl N-[2-[4-(dimethylcarbamoyl)phenyl]ethyl]carbamate (5.9 g, 20.2 mmol, 1.0 equivalent) and DMF (100.0 mL) were added. NaH (1.94 g, 80.8 mmol, 4.0 equivalents) was added portionwise to the above mixture at 0 ° C. The mixture was stirred at room temperature for 30 min, followed by dropwise addition of iodomethane (5.7 g, 40.4 mmol, 2.0 equivalents). The mixture was stirred for another 1 h and then quenched with water at 0 ° C. and extracted with EtOAc (3 x 30 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / EtOAc 1:1) to provide tert-butyl N-[2-[4-(dimethylcarbamoyl)phenyl]ethyl]-N-methylcarbamate (4 g, 64.69%) as a light yellow oil. LCMS (ESI) m / z: [M+H]+ = 251.1

[1232] Step 3: Preparation of N,N-dimethyl-4-[2-(methylamino)ethyl]benzamide

[1233]

[1234] To a 250 mL round-bottom flask containing tert-butyl N-[2-[4-(dimethylcarbamoyl)phenyl]ethyl]-N-methylcarbamate (4.9 g, 16.0 mmol, 1.0 equiv) in DCM (50.0 mL) was added 4M HCl in dioxane (50.0 mL) at 0°C. The resulting mixture was stirred at room temperature for 2 h and then concentrated under vacuum to produce N,N-dimethyl-4-[2-(methylamino)ethyl]benzamide (4 g, quantitative) as a yellow solid. The crude product was used directly in the next step without further purification. LCMS (ESI) m / z: [M+H]+ = 207.3

[1235] Step 4: 4-[2-[(3-amino-6-chloropyridazin-4-yl)(methyl)amino]ethyl]-N,N-dimethylbenzoyl Preparation of amines

[1236]

[1237] To a 250 mL round-bottom flask containing N,N-dimethyl-4-[2-(methylamino)ethyl]benzamide (4.0 g, 19.4 mmol, 1.0 equiv) in ACN (72.0 mL) was added DIEA (12.5 g, 96.9 mmol, 5.0 equiv) and 4-bromo-6-chloropyridazin-3-amine (4.9 g, 23.3 mmol, 1.2 equiv). The resulting mixture was stirred at 100°C under an atmosphere of dry nitrogen for 16 h. The mixture was then cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / EtOAc 1:1) to provide 4-[2-[(3-amino-6-chloropyridazin-4-yl)(methyl)amino]ethyl]-N,N-dimethylbenzamide (2.3 g, 35.53%) as a yellow solid. LCMS (ESI) m / z: [M+H]+ =334.3

[1238] Step 5: 4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl](methyl)amino]ethyl)-N,N-dihydro- Preparation of methylbenzamide

[1239]

[1240] To a solution of 4-[2-[(3-amino-6-chloropyridazin-4-yl)(methyl)amino]ethyl]-N,N-dimethylbenzamide (1.0 g, 3.0 mmol, 1.0 equiv) and 2-hydroxyphenylboronic acid (537 mg, 3.9 mmol, 1.3 equiv) in dioxane (60.0 mL) and water (3.0 mL) was added KCO (1.2 g, 9.0 mmol, 3.0 equiv) and Pd(dppf)Cl (219 mg, 0.3 mmol, 0.1 equiv). After stirring at 80° C. under a nitrogen atmosphere for 2 h, the resulting mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / EtOAc 1:1) to afford 4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl](methyl)amino]ethyl)-N,N-dimethylbenzamide (530 mg, 45.19%) as a brown solid. LCMS (ESI) m / z: [M+H] + = 392.2.

[1241] Step 6: 4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl](methyl)amino]ethyl)benzoic acid Preparation of (I-66)

[1242]

[1243] To a 25 mL round-bottom flask was added 4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl](methyl)amino]ethyl)-N,N-dimethylbenzamide (360 mg, 0.9 mmol, 1.0 equiv) and 12 N HCl (5.0 mL). The resulting mixture was stirred at 70 °C for 16 h, then cooled to room temperature and concentrated under reduced pressure. The crude product was purified by reverse phase preparative HPLC to afford 4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl](methyl)amino]ethyl)benzoic acid (I-66) as a gray solid. , 37 mg, 10.26%). LCMS (ESI) m / z: [M+H] + = 365.3.

[1244] Preparation of 4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)(ethyl)amino)ethyl)benzoic acid (K-79)

[1245]

[1246] Step 1: Preparation of tert-butyl N-[2-(4-cyanophenyl)ethyl]carbamate

[1247]

[1248] To 3-(4-cyanophenyl)propionic acid (2.00 g, 11.416 mmol, 1.00 equiv) was added at room temperature. t To a stirred solution of 1-BuOH (20.00 mL) was added DPPA (6.28 g, 22.820 mmol, 2.00 equiv) and TEA (2.31 g, 22.833 mmol, 2.00 equiv). The resulting mixture was stirred at 60°C overnight. The resulting mixture was concentrated under reduced pressure and then washed with citric acid and subsequently with brine. The organic phase was separated and dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography eluting with PE / EtOAc (4:1) to provide tert-butyl N-[2-(4-cyanophenyl)ethyl]carbamate (1.5 g, 50.68%) as a white solid. LCMS (ESI) m / z [M+H] + =247.

[1249] Step 2: Preparation of tert-butyl N-[2-(4-cyanophenyl)ethyl]-N-ethylcarbamate

[1250]

[1251] To a stirred solution of N-[2-(4-cyanophenyl)ethyl]t-butyl carbamate (1.50 g, 6.090 mmol, 1.00 equivalent) in DMF (20.00 mL) was added iodoethane (1.14 g, 7.309 mmol, 1.20 equivalents) and NaH (0.49 g, 12.251 mmol, 2.01 equivalents, 60%) at 0 ° C. The resulting mixture was stirred at room temperature for 2 h. The reaction was quenched at room temperature by adding frozen water (100 mL). The resulting mixture was extracted with EtOAc (3 x 100 mL). The combined organic layer was washed with salt water (3 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to provide tert-butyl N-[2-(4-cyanophenyl)ethyl]-N-ethylcarbamate (1.1 g, 60.57%) as a white solid, which was used directly in the next step without further purification. LCMS (ESI) m / z: [M+H] + =275.

[1252] Step 3: Preparation of 4-[2-(ethylamino)ethyl]benzonitrile

[1253]

[1254] To a stirred solution of tert-butyl N-[2-(4-cyanophenyl)ethyl]-N-ethylcarbamate (1.10 g, 4.009 mmol, 1.00 equiv) in DCM (10.00 mL) was added TFA (3.67 mL, 49.409 mmol, 12.32 equiv) at room temperature. After stirring at room temperature for 2 h, the resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, MeCN in water (0.2% FA), 0% to 100% gradient in 30 min; detector, UV 254 nm. The product was collected at 10% gradient to provide 4-[2-(ethylamino)ethyl]benzonitrile (600 mg, 77.30%) as a colorless syrup. LCMS (ESI) m / z [M+H ]+ =175.

[1255] Step 4: Preparation of 4-[2-[(3-amino-6-chloropyridazin-4-yl)(ethyl)amino]ethyl]benzonitrile

[1256]

[1257] To a stirred mixture of 4-[2-(ethylamino)ethyl]benzonitrile (600.00 mg, 3.443 mmol, 1.00 equiv) and 4-bromo-6-chloropyridazin-3-amine (1435.48 mg, 6.886 mmol, 2.00 equiv) in DMF (10.00 mL) was added DIEA (1335.10 mg, 10.329 mmol, 3.00 equiv) at room temperature. The resulting mixture was stirred at 100°C overnight. The mixture was diluted with water (100 mL) and extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (3 x 100 mL) and then dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (3:1) to provide 4-[2-[(3-amino-6-chloropyridazin-4-yl)(ethyl)amino]ethyl]benzonitrile (360 mg, 31.87%) as a yellow solid. LCMS (ESI) m / z [M+H ]+ =302.

[1258] Step 5: Preparation of 4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl](ethyl)amino]ethyl)benzonitrile Preparation

[1259]

[1260] To a solution of 4-[2-[(3-amino-6-chloropyridazin-4-yl)(ethyl)amino]ethyl]benzonitrile (360.00 mg, 1.193 mmol, 1.00 equiv) and 2-hydroxyphenylboronic acid (493.62 mg, 3.579 mmol, 3.00 equiv) in dioxane (5.00 mL) and H O (1.00 mL) was added Cs CO (1166.03 mg, 3.579 mmol, 3.00 equiv) and XPhos Pd G (100.97 mg, 0.119 mmol, 0.10 equiv). After stirring at 95 ° C under a nitrogen atmosphere for 4 h, the mixture was cooled to room temperature. The reaction mixture was filtered and concentrated in vacuo. The residue was purified by reverse phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, ACN in water (0.2% FA), 0% to 100% gradient in 10 min; detector, UV 254 nm. The product was collected at 30% gradient to provide 4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl](ethyl)amino]ethyl)benzonitrile (320 mg, 67.17%) as a yellow solid. LCMS (ESI) m / z [M+H ]+ =360.

[1261] Step 6: 4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl](ethyl)amino]ethyl)benzoic acid Preparation of (K-79)

[1262]

[1263] To a stirred mixture of 4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl](ethyl)amino]ethyl)benzonitrile (120.00 mg, 0.334 mmol, 1.00 equiv) in HO (1.00 mL) and MeOH (1.00 mL) was added KOH (32.41 mg, 0.578 mmol, 1.73 equiv) portionwise. The resulting mixture was stirred at 80°C for 2 h. The mixture was cooled to room temperature. The reaction mixture was filtered through a short pad of celite and concentrated in vacuo. The residue was purified by reverse phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, ACN in water (0.2% FA), gradient from 0% to 100% over 30 min; detector, UV 254 nm. This afforded 4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl](ethyl)amino]ethyl)benzoic acid (61 mg, 45.87%) as a white solid. LCMS (ESI) m / z [M+H ]+ =379.

[1264] Preparation of 2-[6-amino-5-[4-(2-aminoethyl)piperazin-1-yl]pyridazin-3-yl]phenol (I-67)

[1265]

[1266] Step 1: Preparation of tert-butyl N-[2-[4-(3-amino-6-chloropyridazin-4-yl)piperazin-1-yl]ethyl]carbamate preparation

[1267]

[1268] To a solution of 3-amino-4-bromo-6-chloropyridazine (4.16 g, 19.958 mmol, 1.00 equiv) in DMF (25.00 mL) was added tert-butyl N-[2-(piperazin-1-yl)ethyl]carbamate (9.15 g, 0.040 mmol, 2 equiv) and DIEA (9.90 mL, 56.837 mmol, 2.85 equiv). The mixture was stirred at 120° C. overnight. After cooling to room temperature, the solvent was removed under vacuum. The residue was purified by silica gel column chromatography eluting with DCM / MeOH (20:1) to provide tert-butyl N-[2-[4-(3-amino-6-chloropyridazin-4-yl)piperazin-1-yl]ethyl]carbamate (2.1329 g, 29.95%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 357.

[1269] Step 2: N-(2-[4-[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]piperazin-1-yl]ethyl)amino Preparation of tert-butyl ester

[1270]

[1271] To a solution of tert-butyl N-[2-[4-(3-amino-6-chloropyridazin-4-yl)piperazin-1-yl]ethyl]carbamate (1.98 g, 5.548 mmol, 1.00 equiv) in dioxane / water (4:1, 50 mL) was added (2-hydroxyphenyl)boronic acid (1.15 g, 8.338 mmol, 1.50 equiv), K2CO3 (3 equiv, 2.3 g), and XPhos Pd G3 (0.1 equiv, 0.555 mmol, 469.64 mg). The resulting mixture was stirred at 80°C under an atmosphere of dry nitrogen. After cooling to room temperature, the solvent was removed under vacuum. The residue was applied to a silica gel column and eluted with DCM / MeOH (30:1) to afford tert-butyl N-(2-[4-[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]piperazin-1-yl]ethyl)carbamate (1.325 g, 57.6%) as a yellow solid. LCMS (ESI) m / z: [M+H] + = 415.

[1272] Step 3: Preparation of 2-[6-amino-5-[4-(2-aminoethyl)piperazin-1-yl]pyridazin-3-yl]phenol (I-67) Preparation

[1273]

[1274] A solution of tert-butyl N-(2-[4-[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]piperazin-1-yl]ethyl)carbamate (1.3 g, 3.316 mmol, 1.00 equiv) in TFA / DCM (1:5, 108 mL) was stirred at room temperature overnight. After removing the solvent under vacuum, the residue was purified by reverse phase flash chromatography. This provided I-67 (0.497 g, 50.4%) as a yellow solid, which was used without further purification. LCMS (ESI) m / z: [M+H] + = 315.

[1275] Preparation of 2-(6-amino-5-(4-((methylamino)methyl)phenethoxy)pyridazin-3-yl)phenol (I-68)

[1276]

[1277] Step 1: Preparation of 2-(4-((methylamino)methyl)phenyl)ethan-1-ol

[1278]

[1279] To a stirred solution of tert-butyl N-[[4-(2-hydroxyethyl)phenyl]methyl]carbamate (1.00 g, 3.979 mmol, 1.00 equiv) in THF (40 mL) was added LiAlH4 (906.09 mg, 23.873 mmol, 6.00 equiv) portionwise at 0 °C under an atmosphere of dry nitrogen. The resulting mixture was stirred at 80 °C for 4 h, then cooled to room temperature, diluted with DCM (30 mL), and quenched with water. The resulting mixture was concentrated under reduced pressure and the residue was purified by reverse phase flash chromatography to afford 2-(4-((methylamino)methyl)phenyl)ethan-1-ol (1.3 g) as an off-white solid. LCMS (ESI) m / z [M+H] + = 166.

[1280] Step 2: Preparation of tert-butyl (4-(2-hydroxyethyl)benzyl)(methyl)carbamate

[1281]

[1282] To a stirred mixture of 2-(4-((methylamino)methyl)phenyl)ethane-1-ol (800.00 mg, 4.842 mmol, 1.00 equivalent) and TEA (979.83 mg, 9.683 mmol, 2.00 equivalent) in DCM (50 mL) was dropwise added tert-butyl dicarbonate (1585.00 mg, 7.262 mmol, 1.50 equivalent) at room temperature under an atmosphere of dry nitrogen. The resulting mixture was stirred overnight and then concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography to provide tert-butyl (4-(2-hydroxyethyl)benzyl)(methyl)carbamate (380 mg, 22.18%) as a white solid. LCMS (ESI) m / z [M+H] + = 266.

[1283] Step 3: tert-Butyl (4-(2-((3-amino-6-chloropyridazin-4-yl)oxy)ethyl)benzyl)(methyl)carbamate Preparation of esters

[1284]

[1285] To a stirred mixture of 4-bromo-6-chloropyridazin-3-amine (226.86 mg, 1.088 mmol, 0.76 equiv) and tert-butyl (4-(2-hydroxyethyl)benzyl)(methyl)carbamate (380.00 mg, 1.432 mmol, 1.00 equiv) in DMF (10 mL) was added NaH (51.89 mg, 2.162 mmol, 1.51 equiv) portionwise at room temperature under an atmosphere of dry nitrogen. The resulting mixture was stirred at 50 ° C for 3 h, then cooled to room temperature and quenched with water (20 mL). The mixture was diluted with EtOAc (10 mL) and washed with brine (2 x 10 mL). The organic layer was dried over Na SO , filtered, and concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography to afford tert-butyl (4-(2-((3-amino-6-chloropyridazin-4-yl)oxy)ethyl)benzyl)(methyl)carbamate as a brown solid (150 mg, 26.66%). LCMS (ESI) m / z [M+H] + = 393.

[1286] Step 4: (4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)oxy)ethyl)benzyl)(methyl)amino Preparation of tert-butyl formate

[1287]

[1288] To a solution of tert-butyl (4-(2-((3-amino-6-chloropyridazin-4-yl)oxy)ethyl)benzyl)(methyl)carbamate (100.00 mg, 0.255 mmol, 1.00 equiv) and 2-hydroxyphenylboronic acid (52.66 mg, 0.382 mmol, 1.50 equiv) in dioxane (2 mL) and water (0.5 mL) was added KCO (70.36 mg, 0.509 mmol, 2.00 equiv) and Pd(PPh) (29.41 mg, 0.025 mmol, 0.10 equiv). After stirring at 100 ° C. under an atmosphere of dry nitrogen for 4 h, the resulting mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by preparative TLC (DCM / MeOH 10:1) to afford tert-butyl (4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)oxy)ethyl)benzyl)(methyl)carbamate (140 mg) as a brown solid. LCMS (ESI) m / z [M+H] + =451.

[1289] Step 5: Reaction of 2-(6-amino-5-(4-((methylamino)methyl)phenethoxy)pyridazin-3-yl)phenol (I-68) preparation

[1290]

[1291] To a stirred solution of tert-butyl (4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)oxy)ethyl)benzyl)(methyl)carbamate (140.00 mg, 0.311 mmol, 1.00 equiv) in DCM (3.0 mL) was added TFA (1.00 mL) dropwise at room temperature under an atmosphere of dry nitrogen. The resulting mixture was stirred for 1 h, then concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography to afford I-68 (80 mg, 73.50%) as a brown oil. LCMS (ESI) m / z [M+H] = 351.

[1292] Preparation of 4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)(methyl)amino)ethyl)benzaldehyde (K-80)

[1293]

[1294] Step 1: Preparation of 4-(2-((3-amino-6-chloropyridazin-4-yl)(methyl)amino)ethyl)benzaldehyde

[1295]

[1296] To a stirred mixture of 4-[2-[(3-amino-6-chloropyridazin-4-yl)(methyl)amino]ethyl]benzonitrile (200.00 mg, 0.695 mmol, 1.00 equiv) in anhydrous DCM (3.0 mL) at 0°C under a nitrogen atmosphere was added DIBAL-H (0.97 mL, 1.5 M in toluene, 1.459 mmol, 2.10 equiv) dropwise. The resulting mixture was stirred at room temperature for 1 h. The mixture was acidified to pH 3 with HCl (0.5 M) at 0°C and then neutralized to pH 7 with saturated NaHCO (aq). The resulting mixture was extracted with DCM (3 x 100 mL). The combined organic layers were washed with brine (3 x 100 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure to give 4-(2-((3-amino-6-chloropyridazin-4-yl)(methyl)amino)ethyl)benzaldehyde (162 mg, 75.84%) as a white solid. The material was used directly in the next step without further purification. LCMS (ESI) m / z: [M+H] + = 291.

[1297] Step 2: 4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)(methyl)amino)ethyl)benzaldehyde Preparation of (K-80)

[1298]

[1299] To a stirred mixture of 4-(2-((3-amino-6-chloropyridazin-4-yl)(methyl)amino)ethyl)benzaldehyde (237.20 mg, 1.720 mmol, 5.00 equiv) in dioxane (5.00 mL) and H O (1.00 mL) was added Pd(dtbpf)Cl (22.42 mg, 0.034 mmol, 0.10 equiv) and Cs CO (448.25 mg, 1.376 mmol, 4.00 equiv). The resulting mixture was stirred at 100 ° C under a nitrogen atmosphere overnight. The mixture was cooled to room temperature, then filtered through a short pad of celite and concentrated in vacuo. The residue was purified by reverse-phase flash chromatography under the following conditions (column, C18 silica gel; mobile phase, ACN in water (0.1% FA), gradient from 0% to 60% over 40 minutes; detector, UV 254 nm). This afforded 4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)(methyl)amino)ethyl)benzaldehyde (29 mg, 20.57%) as a yellow solid. LCMS (ESI) m / z [M+H]+ = 349.

[1300] Preparation of 4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)amino)ethyl)benzaldehyde (K-81)

[1301]

[1302] Step 1: Preparation of methyl 4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)amino)ethyl)benzoate Preparation

[1303]

[1304] In a 40-mL vial purged with nitrogen and maintained under an inert atmosphere of nitrogen, methyl 4-[2-[(3-amino-6-chloropyridazin-4-yl)amino]ethyl]benzoate (250.00 mg, 0.815 mmol, 1.00 equiv), 2-hydroxyphenylboronic acid (168.62 mg, 1.222 mmol, 1.5 equiv), Xphos Pd G3 (68.99 mg, 0.081 mmol, 0.1 equiv), K2CO3 (225.27 mg, 1.630 mmol, 2 equiv), H2O (1.00 mL), and 1,4-dioxane (10.00 mL) were placed. The resulting mixture was stirred at 80°C for 12 h. The mixture was diluted with water (30 mL) and extracted with 2 x 50 mL of ethyl acetate. The organic layers were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by flash preparative HPLC (IntelFlash-1) using the following conditions: column, silica gel; mobile phase, increasing from ACN = 0 to ACN = 30 over 40 min; detector: 254 nm. This afforded 200 mg (67.34%) of methyl 4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)amino)ethyl)benzoate as a yellow solid. LCMS (ESI) m / z: [M+H] + =365.

[1305] Step 2: Preparation of 2-(6-amino-5-((4-(hydroxymethyl)phenethyl)amino)pyridazin-3-yl)phenol

[1306]

[1307] In an 8-mL sealed test tube was placed methyl 4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)amino)ethyl)benzoate (150.00 mg, 0.412 mmol, 1.00 equiv) and THF (2.00 mL) and the vessel was cooled to 0°C. LAH (31.25 mg, 0.823 mmol, 2 equiv) was added at 0°C. The resulting solution was stirred at 0°C for 1 h. The reaction was quenched by the addition of 0.3 mL of water and 0.9 mL of 10% aqueous NaOH. The solid was removed by filtration and the filtrate was concentrated under vacuum. This produced 110 mg (crude) of 2-(6-amino-5-((4-(hydroxymethyl)phenethyl)amino)pyridazin-3-yl)phenol as an off-white solid. LCMS (ESI) m / z: [M+H] + =337.

[1308] Step 3: Preparation of 4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)amino)ethyl)benzaldehyde (K-81) preparation

[1309]

[1310] In a 50-mL round-bottom flask was placed 2-(6-amino-5-((4-(hydroxymethyl)phenethyl)amino)pyridazin-3-yl)phenol (90.00 mg, 0.268 mmol, 1.00 equiv), DCM (20.00 mL) and MnO2 (465.19 mg, 5.351 mmol, 20 equiv). The resulting solution was stirred at room temperature for 24 h. The solid was filtered off. The filtrate was concentrated under vacuum and the residue was purified on a silica gel column with ethyl acetate / petroleum ether (4:1). This yielded 25 mg (27.95%) of 4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]amino]ethyl)benzaldehyde as a dark yellow solid. LCMS (ESI) m / z: [M+H] + =335.

[1311] Preparation of 4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)amino)ethyl)benzoic acid (K-82)

[1312]

[1313] Step 1: Preparation of methyl 4-(2-aminoethyl)benzoate

[1314]

[1315] To a solution of 4-[2-[(tert-butoxycarbonyl)amino]ethyl]benzoic acid (3.00 g, 11.308 mmol, 1.00 equiv) in MeOH (20.00 mL) was added H2SO4 (2.50 mL, 46.901 mmol, 4.15 equiv) and the resulting solution was stirred at 25 °C for 20 hours. The resulting mixture was concentrated under reduced pressure and diluted with water (20 mL). The mixture was adjusted to pH 7-8 with saturated sodium bicarbonate solution and extracted with CHCl (50 mL x 3). The organic layers were combined and dried over anhydrous sodium sulfate, filtered, and concentrated to give methyl 4-(2-aminoethyl)benzoate (1.4 g, 69.08%) as a brown solid, which was used in the next step without further purification. LCMS (ESI) m / z: [M+H] + =180.

[1316] Step 2: Preparation of methyl 4-[2-[(3-amino-6-chloropyridazin-4-yl)amino]ethyl]benzoate

[1317]

[1318] To a solution of methyl 4-(2-aminoethyl)benzoate (1.40 g, 7.812 mmol, 1.00 eq) and 4-bromo-6-chloropyridazin-3-amine (1.95 g, 9.374 mmol, 1.2 eq) in DMF (10 mL) was added DIEA (2.02 g, 15.623 mmol, 2 eq). The resulting solution was stirred at 110 ° C for 15 hours. The mixture was cooled to room temperature, then filtered through a short pad of celite and the filtrate was concentrated in vacuo. The residue was purified by flash C18 chromatography eluting with a 0-26% gradient of ACN in H2O to produce methyl 4-[2-[(3-amino-6-chloropyridazin-4-yl)amino]ethyl]benzoate (305 mg, 12.73%) as a yellow solid. LCMS (ESI) m / z: [M+H] + =307.

[1319] Step 3: Preparation of methyl 4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]amino]ethyl)benzoate Preparation

[1320]

[1321] To a solution of 4-[2-[(3-amino-6-chloropyridazin-4-yl)amino]ethyl]benzoic acid methyl ester and 2-hydroxyphenylboronic acid (70.0 mg, 0.652 mmol, 2 equivalents) in dioxane (4.00 mL) and H O (1.00 mL) was added Xphos Pd G (27.6 mg, 0.033 mmol, 0.1 equivalent) and K CO (90.1, 0.652 mmol, 2 equivalents). The resulting solution was stirred at 80° C. for 3 hours. The mixture was diluted with water (30 mL) and extracted with EtOAc (30 mL x 3). The organic layers were combined and dried over anhydrous sodium sulfate, filtered and concentrated to produce a crude product. The crude product was purified by preparative TLC (CH2Cl2 / MeOH 10:1) to give methyl 4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]amino]ethyl)benzoate (75 mg, 63.13%) as a yellow solid. LCMS (ESI) m / z: [M+H] + =365.

[1322] Step 4: Reaction of 4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]amino]ethyl)benzoic acid (K-82) preparation

[1323]

[1324] To a solution of methyl 4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]amino]ethyl)benzoate (55.0 mg, 0.151 mmol, 1.00 equiv) in THF (0.50 mL) and H O (0.10 mL) was added LiOH (36.2 mg, 1.509 mmol, 10 equiv) and the resulting solution was stirred at 25 ° C for 15 hours. The mixture was diluted with water (10 mL) and extracted with EtOAc (10 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by flash C18 chromatography eluting with a 0-26% ACN gradient in H O to produce 4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]amino]ethyl)benzoic acid (32 mg, 60.51%) as a yellow solid. LCMS (ESI) m / z: [M+H] + =351.

[1325] Preparation of 2-(6-amino-5-((4-(aminomethyl)phenethyl)amino)pyridazin-3-yl)phenol (K-83)

[1326]

[1327] Step 1: Preparation of N4-[2-[4-(aminomethyl)phenyl]ethyl]-6-chloropyridazine-3,4-diamine

[1328]

[1329] To a solution of 4-[2-[(3-amino-6-chloropyridazin-4-yl)amino]ethyl]benzamide (100.00 mg, 0.343mmol, 1.00 equivalent) in THF (3 mL) was added LiAlH4 (130.10 mg, 3.428 mmol, 10 equivalents). The resulting solution was stirred at 25 ° C for 5 hours. The mixture was carefully quenched with water (5 mL), diluted with EtOAc (50 mL), and washed with water (50 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated to produce a crude product. The crude product was purified by flash C18 chromatography, eluting with a 0-22% ACN gradient in H2O to produce N4-[2-[4-(aminomethyl)phenyl]ethyl]-6-chloropyridazine-3,4-diamine (37 mg, 38.86%) as a white solid. LCMS (ESI) m / z: [M+H] + =278.

[1330] Step 2: 2-[6-amino-5-([2-[4-(aminomethyl)phenyl]ethyl]amino)pyridazin-3-yl]phenol (K- Preparation of 83)

[1331]

[1332] To a stirred solution of N4-[2-[4-(aminomethyl)phenyl]ethyl]-6-chloropyridazine-3,4-diamine (37.00 mg, 0.133 mmol, 1.00 equiv) and 2-hydroxyphenylboronic acid (36.75 mg, 0.266 mmol, 2 equiv) in dioxane (1.6 mL) and H2O (0.4 mL) was added Xphos Pd G3 (11.28 mg, 0.013 mmol, 0.1 equiv) and K2CO3 (36.82 mg, 0.266 mmol, 2.00 equiv). The resulting mixture was stirred at 80 ° C for 2 h. The mixture was cooled to room temperature. The reaction mixture was filtered through a short pad of celite and concentrated in vacuo. The residue was purified by flash C18 chromatography eluting with a 0-8% gradient of ACN in H2O to give 2-[6-amino-5-([2-[4-(aminomethyl)phenyl]ethyl]amino)pyridazin-3-yl]phenol as a brown solid (19 mg, 42.52%). LCMS (ESI) m / z: [M+H] + =336.

[1333] Preparation of 4-(4-(2-((4-(3-(3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)pyridin-2-yl)oxy)ethyl)piperazin-1-yl)butanoic acid (K-84)

[1334]

[1335] Step 1: 5-(4-[2-[(4-[3-[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]-3,8-diazabicyclo Preparation of [3.2.1] methyl [octan-8-yl]pyridin-2-yl]oxy]ethyl]piperazin-1-yl]pentanoate

[1336] To a stirred solution of 2-[6-amino-5-(8-[2-[2-(piperazin-1-yl)ethoxy]pyridin-4-yl]-3,8-diazabicyclo[3.2.1]octan-3-yl)pyridazin-3-yl]phenol (20 mg, 0.040 mmol, 1.00 equiv) and methyl 4-oxobutanoate (9.24 mg, 2.00 equiv) in MeOH (2.00 mL) was added AcOH (0.03 mL, 0.040 mmol) and NaBH3CN (12.50 mg, 0.200 mmol, 5.00 equiv) at room temperature. The resulting mixture was stirred for 2 h. The reaction was quenched with H2O at room temperature. The residue was purified by reverse phase flash chromatography to provide the title compound (8.9 mg, 36.26%). LCMS (ESI) m / z: [M+H] + = 603.

[1337] Step 2: 4-(4-(2-((4-(3-(3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)-3,8-diazabicyclo Preparation of [3.2.1] (octane-8-yl)pyridin-2-yl)oxy)ethyl)piperazin-1-yl)butanoic acid (K-84)

[1338]

[1339] To a stirred mixture of methyl 5-(4-[2-[(4-[3-[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]-3,8-diazabicyclo[3.2.1]octan-8-yl]pyridin-2-yl)oxy]ethyl]piperazin-1-yl)pentanoate (8.00 mg, 0.013 mmol, 1.00 equiv) in MeOH (0.60 mL) and HO (0.30 mL) was added LiOH (3.18 mg, 0.130 mmol, 10.00 equiv) at room temperature. After 4 h, the mixture was neutralized to pH 7 with HCl (1 M). The resulting mixture was concentrated under reduced pressure to afford the title compound, which was used in the next step without further purification. LCMS (ESI) m / z: [M+H] + = 589.

[1340] The following intermediates in Table A16 were prepared in a similar manner as described in the preparation of K-84.

[1341]

[1342] Preparation of 2-(6-amino-5-(8-(2-((1r,3r)-3-(piperidin-4-yloxy)cyclobutyloxy)pyridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-3-yl)pyridazin-3-yl)phenol (K-86)

[1343]

[1344] Step 1: 4-((1r,3r)-3-((4-(3-(3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)-3,8-diazo Preparation of tert-butyl heterobicyclo[3.2.1]octan-8-yl)pyridin-2-yl)oxy)cyclobutyloxy)piperidine-1-carboxylate

[1345]

[1346] To a stirred solution of tert-butyl 4-[(1r,3r)-3-([4-[3-(3-amino-6-chloropyridazin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl]pyridin-2-yl]oxy)cyclobutyloxy]piperidine-1-carboxylate (200.00 mg, 0.341 mmol, 1.00 equiv) and 2-hydroxyphenylboronic acid (141.19 mg, 1.023 mmol, 3.00 equiv) in 1,4-dioxane (8.00 mL) and H2O (2.00 mL) at room temperature was added XPhosPdG3 (57.77 mg, 0.068 mmol, 0.20 equiv) and Cs2CO3 (333.53 mg, 1.023 mmol, 3.00 equiv). The resulting mixture was stirred at 100° C. for 1 h under a nitrogen atmosphere. The mixture was cooled to room temperature. The residue was purified by reverse phase flash chromatography to provide the title compound (224 mg, crude) as a brown solid. LCMS (ESI) m / z: [M+H] + = 644.

[1347] Step 2: 2-[6-amino-5-(8-[2-[(1r,3r)-3-(piperidin-4-yloxy)cyclobutyloxy]pyridine-4-yloxy] Preparation of 3,8-diazabicyclo[3.2.1]octan-3-yl)pyridazin-3-yl]phenol (K-86)

[1348]

[1349] To a stirred solution of tert-butyl 4-((1r,3r)-3-((4-(3-(3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)pyridin-2-yl)oxy)cyclobutyloxy)piperidine-1-carboxylate (224.00 mg, 0.348 mmol, 1.00 equiv) in DCM (6.00 mL) was added TFA (3.00 mL, 40.389 mmol, 116.08 equiv) at room temperature. The resulting mixture was stirred at room temperature for 1 h and then concentrated under reduced pressure to afford the title compound (373 mg, crude) as a brown oil. LCMS (ESI) m / z: [M+H] + = 544.

[1350] Preparation of 2-(6-amino-5-(4-(piperazin-1-ylmethyl)phenethoxy)pyridazin-3-yl)phenol (K-87)

[1351]

[1352] Step 1: 4-(4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)oxy)ethyl)benzyl)piperazine- Preparation of tert-butyl 1-formate

[1353]

[1354] To a stirred solution of 4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]oxy]ethyl)benzaldehyde (80.00 mg, 0.24 mmol, 1.00 equiv) and tert-butyl piperazine-1-carboxylate (66.72 mg, 0.0368 mmol, 1.50 equiv) in a mixture of CHCl (4 mL) and methanol (4 mL) was added acetic acid until the solution reached pH 6. NaBHCN (60.00 mg, 0.960 mmol, 4.00 equiv) was then added portionwise at room temperature. The resulting mixture was stirred for 2 h and then concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography to afford the title compound (60 mg, 53.27%) as a white solid. LCMS (ESI) m / z: [M+H] + =506.

[1355] Step 2: Preparation of 2-(6-amino-5-(4-(piperazin-1-ylmethyl)phenethoxy)pyridazin-3-yl)phenol (K-87) preparation

[1356]

[1357] To a stirred solution of tert-butyl 4-(4-(2-((3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)oxy)ethyl)benzyl)piperazine-1-carboxylate (60.00 mg, 0.12 mmol, 1.00 equiv) in DCM (6.00 mL) was added TFA (2.00 mL) dropwise at room temperature. The resulting mixture was stirred for 2 h and then concentrated in vacuo. The crude product mixture was used directly in the next step without further purification. This produced the title compound (68.00 mg, crude) as a brown oil. LCMS (ESI) m / z: [M+H] + =405.

[1358] The following intermediates in Table A17 were prepared in a similar manner as described in the preparation of K-87.

[1359]

[1360] Example 2. Preparation of N-(2-[[2-(2-[4-[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]piperazin-1-yl]acetylamino)ethyl](methyl)amino]ethyl)-2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-4-yl]oxy]acetamide (Compound 104)

[1361]

[1362] To a stirred mixture of 2-(4-(3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl)piperazin-1-yl)acetic acid (I-62, 36.2 mg, 0.083 mmol, 1.20 equiv) in DMF (1 mL) at room temperature under an atmosphere of dry nitrogen are added DIEA (49.05 mg, 0.380 mmol, 5.00 equiv) and HATU (37.52 mg, 0.099 mmol, 1.10 equiv), and the resulting mixture is stirred for 30 min. To the above mixture was added N-(2-((2-aminoethyl)(methyl)amino)ethyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetamide (I-63, 36.02 mg, 0.083 mmol, 1.10 equiv) portionwise. The resulting mixture was stirred at room temperature for another 1 h. The reaction was quenched with water (5 mL) at room temperature, and the resulting mixture was extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (10 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by reverse phase preparative HPLC to afford Compound 104 (4.7 mg, 8.00%) as a yellow solid. 1 H NMR (300 MHz, DMSO- d 6 ) δ 11.12 (s, 1H), 8.14 (s, 1H), 7.97 (d, J = 5.9 Hz, 1H), 7.89 (d, J = 8.0 Hz, 1H), 7.78 (t, J = 7.8 Hz, 2H), 7.48 (d, J = 6.6 Hz, 2H), 7.38 (d, J = 8.6 Hz, 1H), 7.24 (t, J = 7.8 Hz, 1H), 6.89 (d, J = 7.7 Hz, 2H), 6.25 (s,2H), 5.12 (dd, J = 12.9, 5.3 Hz, 1H), 4.78 (s, 2H), 3.14 (s, 7H), 3.00 - 2.90(m, 5H), 2.62 -2.50 (m, 9H), 2.30 (s, 3H), 2.04 (dd, J= 12.7, 6.5 Hz, 1H). LCMS (ESI) m / z [M+H] + = 743.55.

[1363] Using procedures similar to those used to prepare compound 104, the following compounds in Table B were prepared.

[1364] Table B

[1365]

[1366]

[1367]

[1368]

[1369] Example 3. Preparation of 4-(2-[[3-amino-6-(2-hydroxyphenyl)pyridazin-4-yl]oxy]ethyl)...

Claims

1. A compound having a structure of Formula I: in X 1 Is O or NR 2 ; Each X 2 are independently halogen; k is 0 or 1; m is 0; R 1 is halogen or C1-C6 alkyl; R 2 is H or C1-C6 alkyl; L 1 is C1-C6 alkylene; L is included structural joints; n is 0, 1, 2, or 3; L 2 is an optionally substituted C1-C6 alkylene group, an optionally substituted C1-C 20 heteroalkylene or optionally substituted C3-C9 heterocyclylene; Each L 3 is independently -O-, optionally substituted C1-C 20 heteroalkylene, optionally substituted C3-C 10 Carbocyclylene, optionally substituted C3-C 10 Carbocyclylene-C1-C 20 alkylene, optionally substituted C3-C9 heterocyclylene or optionally substituted C3-C9 heterocyclylene-C1-C 20 Alkylene; Where L 2 and L 3 The "optionally substituted" in the definition of means optionally substituted by oxo; and D is the degradation part, Wherein the degradation part comprises the structure of formula A: in Y 1 yes R A5 is H, C1-C6 alkyl; R A6 is H; and R A7 It is H; R A8 is H, C1-C6 alkyl; R A1 , R A2 , R A3 and R A4 Each of which is independently H, A 2 ;and A 2 is the bond between the degradation moiety and the linker; or Wherein the degradation part has the structure of formula C: in L 6 Yes-N(R B1 )(R B2 ), R B1 It is H, A 2 , optionally substituted C1-C6 alkyl; R B2 is H, optionally substituted C1-C6 alkyl; R B3 Yes A 2 , C1-C6 alkyl; R B4 It is H; R B5 It is H; v2 is 0, 1; Each R B6 Independently is A 2 ; R B7 and R B8 Each of R is independently H, halogen, C1-C6 alkyl; B9 is H or C1-C6 alkyl; and A 2 is the bond between the degradable moiety and the linker; Where R B1 , R B3 and R B6 One and only one of them is A 2 ;or wherein the degradation portion comprises The structure of A 2 is the bond between the degradable moiety and the linker; or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1, wherein X 1 It's O.

3. The compound according to claim 1 or 2, wherein L 1 yes 4. The compound according to claim 1 or 2, wherein L 2 yes 5. The compound according to claim 1 or 2, wherein L 2 yes 6. The compound of claim 1 or 2, wherein each L 3 is independently optionally substituted C3-C 10 Carbocyclylene-C1-C6 alkylene, optionally substituted C3-C9 heterocyclylene or optionally substituted C3-C9 heterocyclylene-C1-C6 alkylene, wherein the "optionally substituted" means optionally substituted by oxo.

7. The compound of claim 1 or 2, wherein each L 3 Independently 8. The compound of claim 1 or 2, wherein n is 0.

9. The compound of claim 1 or 2, wherein R A5 is H or 10. The compound of claim 1 or 2, wherein R A1 , R A2 , R A3 and R A4 Each of is independently H or A 2 ;or R A1 Yes A 2 And R A2 , R A3 and R A4 Each of is H; or R A2 Yes A 2 And R A1 , R A3 and R A4 Each of is H; or R A3 Yes A 2 And R A1 , R A2 and R A4 Each of is H; or R A4 Yes A 2 And R A1 , R A2 and R A3 Each of is H.

11. The compound of claim 1 or 2, wherein Y 1 yes 12. The compound of claim 1 or 2, wherein Y 1 yes 13. The compound of claim 1 or 2, wherein the degradation moiety comprises a structure of formula A2: or a pharmaceutically acceptable salt thereof.

14. The compound of claim 1 or 2, wherein the degradation moiety comprises a structure of formula A4: or a pharmaceutically acceptable salt thereof.

15. The compound of claim 1 or 2, wherein the degradation moiety comprises a structure of formula A5: or a pharmaceutically acceptable salt thereof.

16. The compound of claim 1 or 2, wherein the degradation moiety comprises a structure of formula A6: or a pharmaceutically acceptable salt thereof.

17. The compound of claim 1 or 2, wherein the degradation moiety comprises a structure of formula A8: or a pharmaceutically acceptable salt thereof.

18. The compound of claim 1 or 2, wherein the degradation portion comprises a structure of formula A10: or a pharmaceutically acceptable salt thereof.

19. The compound of claim 1 or 2, wherein the degradation portion comprises structure.

20. The compound of claim 1 or 2, wherein the degradation portion comprises structure.

21. The compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein the degradation portion has a structure of Formula C1:

22. The compound of claim 21 or a pharmaceutically acceptable salt thereof, wherein the degradation portion has a structure of Formula C2:

23. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R B9 It is a C1-C6 alkyl group.

24. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R B9 Bonded to the (S)-stereocenter.

25. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R B9 It's hydrogen.

26. The compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein the degradation portion has the following structure:

27. The compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein the degradation portion has the following structure:

28. The compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein the degradation portion has the following structure:

29. The compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein the degradation portion has the following structure:

30. The compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein the degradation portion is 31. A compound selected from Compounds 1 to 75 in Table 1 and a pharmaceutically acceptable salt thereof:

32. A compound selected from the group consisting of compounds 105-267 in Table 2 and pharmaceutically acceptable salts thereof:

33. A compound selected from compounds 76-104 in Table 2 or a pharmaceutically acceptable salt thereof:

34. A pharmaceutical composition comprising a compound according to any one of claims 1 to 33 and a pharmaceutically acceptable excipient.

35. Use of a compound according to any one of claims 1 to 33 or a pharmaceutical composition according to claim 34 for the preparation of a medicament for treating a BAF complex-associated disorder in a subject in need thereof, wherein the BAF complex-associated disorder is colorectal cancer, breast cancer and pancreatic cancer.

36. Use of a compound according to any one of claims 1 to 33 or a pharmaceutical composition according to claim 34 for the preparation of a medicament for treating a disorder associated with a BRG1 loss-of-function mutation in a subject in need thereof, wherein the disorder associated with a BRG1 loss-of-function mutation is colorectal cancer, breast cancer and pancreatic cancer.

37. Use of a compound according to any one of claims 1 to 33 or a pharmaceutical composition according to claim 34 for the preparation of a medicament for treating cancer in a subject in need thereof, wherein the cancer is selected from colorectal cancer, breast cancer and pancreatic cancer.

38. The use of any one of claims 35-37, wherein the breast cancer is ER-positive breast cancer, ER-negative breast cancer, triple-positive breast cancer, or triple-negative breast cancer.

39. The use of claim 37, wherein the cancer is breast cancer, and the breast cancer is ER-positive breast cancer, ER-negative breast cancer, triple-positive breast cancer, or triple-negative breast cancer.

40. A method for reducing the level and / or activity of BRG1 and / or BRM in a cancer selected from colorectal cancer, breast cancer, and pancreatic cancer for non-therapeutic and non-diagnostic purposes, the method comprising contacting cells of the cancer selected from colorectal cancer, breast cancer, and pancreatic cancer with an effective amount of a compound according to any one of claims 1 to 33 or a pharmaceutical composition according to claim 34.

41. The method of claim 40, wherein the cancer is breast cancer, and the breast cancer is ER-positive breast cancer, ER-negative breast cancer, triple-positive breast cancer, or triple-negative breast cancer.

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