COMPOUNDS AND USES THEREOF

BACKGROUND

Disorders can be affected by the BAF complex. BRD9 is a component of the BAF complex. The present invention relates to useful compositions and methods for the treatment of BAF complex-related disorders, such as cancer and infection.

SUMMARY

Bromodomain-containing protein 9 (BRD9) is a protein encoded by the BRD9 gene on chromosome 5. BRD9 is a component of the BAF (BRG1- or BRM-associated factors) complex, a SWI/SNF ATPase chromatin remodeling complex, and belongs to family IV of the bromodomain-containing proteins. BRD9 is present in several SWI/SNF ATPase chromatin remodeling complexes and is upregulated in multiple cancer cell lines. Accordingly, agents that reduce the levels and/or activity of BRD9 may provide new methods for the treatment of disease and disorders, such as cancer and infection. The inventors have found that depleting BRD9 in cells results in the depletion of the SS18-SSX fusion protein in those cells. The SS18-SSX fusion protein has been detected in more than 95% of synovial sarcoma tumors and is often the only cytogenetic abnormality in synovial sarcoma. Additionally, evidence suggests that the BAF complex is involved in cellular antiviral activities. Thus, agents that degrade BRD9 (e.g., compounds) are useful in the treatment of disorders (e.g., cancers or infections) related to BAF, BRD9, and/or SS18-SSX.

The present disclosure features compounds and methods useful for treating BAF-related disorders (e.g., cancer or infection).

In an aspect, the disclosure features a compound having the structure of Formula I:

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where

R¹is H, optionally substituted C₁-C₆alkyl, optionally substituted C₂-C₆alkenyl, optionally substituted C₁-C₆heteroalkyl, or optionally substituted C₃-C₁₀carbocyclyl;

Z¹is CR²or N;

R²is H, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, or optionally substituted C₂-C₉heteroaryl;

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X¹is CR^X1or N;

X²is O or S;

R^X1is H or optionally substituted C₁-C₆alkyl;

R³is H, cyano, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₁-C₆alkoxy, optionally substituted amino, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₃-C₁₀heterocyclyl, or optionally substituted C₂-C₉heteroaryl; and

G is optionally substituted C₃-C₁₀carbocyclyl, C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, or optionally substituted C₂-C₉heteroaryl, or a pharmaceutically acceptable salt thereof.

In some embodiments,

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In some embodiments,

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In some embodiments,

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In some embodiments,

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In some embodiments,

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In some embodiments,

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In some embodiments, the compound of Formula I has the structure of Formula IA:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula I has the structure of Formula IB:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula I has the structure of Formula IC:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula I has the structure of Formula ID:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula I has the structure of Formula IE:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula I has the structure of Formula IF:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, R¹is H, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, or optionally substituted C₃-C₁₀carbocyclyl. In some embodiments, R¹is H, optionally substituted C₁-C₆alkyl, optionally substituted C₂-C₆alkenyl, or optionally substituted C₃-C₁₀carbocyclyl. In some embodiments, R¹is H, optionally substituted C₁-C₆alkyl, or optionally substituted C₃-C₁₀carbocyclyl.

In some embodiments, R¹is H. In some embodiments, R¹is optionally substituted C₁-C₆alkyl. In some embodiments, R¹is optionally substituted C₂-C₆alkenyl. In some embodiments, R¹is optionally substituted C₃-C₁₀carbocyclyl.

In some embodiments, optionally substituted C₁-C₆alkyl is C₁-C₆perfluoroalkyl.

In some embodiments, R¹is

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In some embodiments, R¹is

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In some embodiments, R¹is

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In some embodiments, R¹is H,

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In some embodiments, R¹is

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In some embodiments, R¹is H,

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In some embodiments, R¹is H,

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In some embodiments, R¹is H,

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In some embodiments, R¹is H or

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In some embodiments, R¹is H. In some embodiments, R¹is

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In some embodiments, Z¹is CR². In some embodiments, Z¹is N.

In some embodiments, R²is H, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₃-C₁₀carbocyclyl, or optionally substituted C₆-C₁₀aryl.

In some embodiments, R²is H, halogen, or optionally substituted C₁-C₆alkyl.

In some embodiments, R²is H, F, or

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In some embodiments, R²is H. In some embodiments, R²is F. In some embodiments, R²is

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In some embodiments, R^X1is H. In some embodiments, R^X1is

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In some embodiments, X¹is CH or N. In some embodiments, X¹is CH. In some embodiments, X¹is N.

In some embodiments, X²is O. In some embodiments, X²is S.

In some embodiments, R³is H, optionally substituted C₁-C₆alkyl, optionally substituted C₂-C₉heteroaryl, or optionally substituted C₁-C₆heteroalkyl.

In some embodiments, R³is H, cyano, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₁-C₆alkoxy, optionally substituted amino, optionally substituted C₃-C₁₀heterocyclyl, or optionally substituted C₂-C₉heteroaryl. In some embodiments, R³is optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, or optionally substituted C₃-C₁₀heterocyclyl. In some embodiments, R³is cyano, optionally substituted C₁-C₆alkoxy, or optionally substituted amino.

In some embodiments, R³is H, optionally substituted C₁-C₆alkyl, or optionally substituted C₁-C₆heteroalkyl. In some embodiments, R³is H or optionally substituted C₁-C₆alkyl. In some embodiments, R³is H. In some embodiments, R³is optionally substituted C₁-C₆alkyl.

In some embodiments, R³is optionally substituted C₂-C₉heteroaryl or optionally substituted C₁-C₆heteroalkyl. In some embodiments, R³is optionally substituted C₁-C₆heteroalkyl.

In some embodiments, R³is

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where

W¹is NR⁴or O;

W²is NH, O, or S;

W³is NR⁷or O;

R⁴is H or optionally substituted C₁-C₆alkyl;

R⁵is optionally substituted C₁-C₆alkyl, optionally substituted C₃-C₁₀carbocyclyl, or optionally substituted C₂-C₉heterocyclyl;

R⁶is H, optionally substituted C₁-C₆alkyl, optionally substituted C₃-C₁₀carbocyclyl, or optionally substituted C₂-C₉heterocyclyl;

R⁷is H or optionally substituted C₁-C₆alkyl; and

R⁸is H, optionally substituted C₁-C₆alkyl, optionally substituted C₃-C₁₀carbocyclyl, or optionally substituted C₂-C₉heterocyclyl.

In some embodiments, R³is optionally substituted C₂-C₉heteroaryl,

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In some embodiments, R³is optionally substituted C₂-C₉heteroaryl. In some embodiments, R³is

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In some embodiments, R³is

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In some embodiments, R³is

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In some embodiments, W¹is NR⁴. In some embodiments, W¹is O.

In some embodiments, W²is NH. In some embodiments, W²is O. In some embodiments, W²is S.

In some embodiments, W³is NR⁷. In some embodiments, W³is O.

In some embodiments, R³is

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In some embodiments, R³is

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In some embodiments, R³is

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In some embodiments, R⁴is H,

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In some embodiments, R⁵is

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In some embodiments, R⁵is

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In some embodiments, R⁶is

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In some embodiments, R⁶is

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In some embodiments, R⁶is H,

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In some embodiments, R⁶is H,

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In some embodiments, R⁷is H,

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In some embodiments, R³is H. In some embodiments, R³is cyano. In some embodiments, R³is

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In some embodiments, R³is

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In some embodiments, R³is

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In some embodiments, R³is

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In some embodiments, R³is

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In some embodiments, R³is

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In some embodiments, R³is

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In some embodiments, G is optionally substituted C₃-C₁₀carbocyclyl or optionally substituted C₂-C₉heterocyclyl. In some embodiments, G is optionally substituted C₆-C₁₀aryl or optionally substituted C₂-C₉heteroaryl.

In some embodiments, G is optionally substituted C₃-C₁₀carbocyclyl. In some embodiments, G is optionally substituted C₆-C₁₀aryl. In some embodiments, G is optionally substituted C₂-C₉heterocyclyl. In some embodiments, G is optionally substituted C₂-C₉heteroaryl.

In some embodiments, G is

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where

each of R^G1, R^G2, R^G3, R^G4, and R^G6is, independently, H, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₂-C₉heteroaryl, optionally substituted C₂-C₆alkenyl, optionally substituted C₂-C₆heteroalkenyl, optionally substituted —O—C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₂-C₅heterocyclyl, hydroxyl, thiol, or optionally substituted amino; or R^G1and R^G2, R^G2and R^G3, R^G3and R^G4, and/or R^G4and R^G6, together with the carbon atoms to which each is attached, combine to form optionally substituted C₆-C₁₀aryl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heteroaryl, or optionally substituted C₂-C₉heterocyclyl.

In some embodiments, each of R^G1, R^G2, R^G3, R^G4, and R^G5is, independently, H, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₂-C₉heteroaryl, optionally substituted C₂-C₆alkenyl, optionally substituted C₂-C₆heteroalkenyl, optionally substituted —O—C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₂-C₅heterocyclyl, hydroxyl, thiol, or optionally substituted amino; or R^G1and R^G2, R^G2and R^G3, R^G3and R^G4, and/or R^G4and R^G5, together with the carbon atoms to which each is attached, combine to form optionally substituted C₆-C₁₀aryl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heteroaryl, or optionally substituted C₂-C₉heterocyclyl.

In some embodiments, each of R^G1, R^G2, R^G3, R^G4, and R^G5is, independently, H, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted —O—C₃-C₆carbocyclyl, or optionally substituted —C₁-C₃alkyl-C₂-C₅heterocyclyl; or R^G1and R^G2, R^G2and R^G3, R^G3and R^G4, and/or R^G4and R^G5, together with the carbon atoms to which each is attached, combine to form optionally substituted C₂-C₉heteroaryl or optionally substituted C₂-C₉heterocyclyl.

In some embodiments, each of R^G1, R^G2, R^G3, R^G4, and R^G5is, independently, H, F, Cl,

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In some embodiments, each of R^G1, R^G2, R^G3, R^G4, and R^G5is, independently, H, F,

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In some embodiments, each of R^G1, R^G2, R^G3, R^G4, and R^G5is, independently, H, F, Cl,

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In some embodiments, R^G1is H; R^G2is

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and R^G5is H. In some embodiments, R^G1is H; R^G2is

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R^G3is

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R^G4is H; and R^G5is

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In some embodiments, R^G1is H; R^G2is

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R^G3is

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R^G4is Cl or F; R^G5is H. In some embodiments, R^G1is H; R^G2is

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R^G3is

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R^G4is H; and R^G5is H. In some embodiments, R^G1is H; R^G2is

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R^G3is

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R^G4is

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and R^G5is H.

In some embodiments, R^G1and R^G2; R^G2and R^G3; R^G3and R^G4, and/or R^G4and R^G5, together with the carbon atoms to which each is attached, combine to form optionally substituted C₂-C₉heterocyclyl. In some embodiments, R^G1and R^G2; R^G2and R^G3; R^G3and R^G4, and/or R^G4and R^G5, together with the carbon atoms to which each is attached, combine to form optionally substituted C₂-C₉heteroaryl.

In some embodiments, G is

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where R^G6is H or optionally substituted C₁-C₆alkyl. In some embodiments, G is

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where R^G6is H or optionally substituted C₁-C₆alkyl.

In some embodiments, G is

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where R^G6is H or optionally substituted C₁-C₆alkyl.

In some embodiments, R^G6is H,

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In some embodiments, R^G6is H or

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In some embodiments, R^G6is H.

In some embodiments, R^G1is H, F,

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In some embodiments, R^G1is H.

In some embodiments, R^G2is H, F,

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In some embodiments, R^G2is H.

In some embodiments, R^G3is H, F,

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In some embodiments, R^G3is H.

In some embodiments, R^G4is H, F,

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In some embodiments, R^G4is H.

In some embodiments, R^G5is H, F,

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In some embodiments, R^G5is H.

In some embodiments, one or more of R^G1, R^G2, R^G3, R^G4, and R^G5is H. In some embodiments, two or more of R^G1, R^G2, R^G3, R^G4, and R^G5is H. In some embodiments, three or more of R^G1, R^G2, R^G3, R^G4, and R^G5is H. In some embodiments, each of R^G1, R^G2, R^G3, R^G4, and R^G5is H.

In some embodiments, G is

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where

each of R^G7, R^G8, R^G9, R^G10, and R^G11is, independently, H, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₂-C₉heteroaryl, optionally substituted C₂-C₆alkenyl, optionally substituted C₂-C₆heteroalkenyl, optionally substituted —O—C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₂-C₅heterocyclyl, hydroxyl, thiol, or optionally substituted amino; or R^G7and R^G8, R^G8and R^G9, R^G9and R^G10, and/or R^G10and R^G11, together with the carbon atoms to which each is attached, combine to form optionally substituted C₆-C₁₀aryl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heteroaryl, or C₂-C₉heterocyclyl.

In some embodiments, each of R^G7, R^G8, R^G9, R^G10, and R^G11is, independently, H, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₂-C₉heteroaryl, optionally substituted C₂-C₆alkenyl, optionally substituted C₂-C₆heteroalkenyl, hydroxyl, thiol, or optionally substituted amino; or R^G7and R^G8, R^G8and R^G9, R^G9and R^G10, and/or R^G10and R^G11, together with the carbon atoms to which each is attached, combine to form optionally substituted C₆-C₁₀aryl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heteroaryl, or C₂-C₉heterocyclyl.

In some embodiments, each of R^G7, R^G8, R^G9, R^G10, and R^G11is, independently, H, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted —O—C₃-C₆carbocyclyl, or optionally substituted —C₁-C₃alkyl-C₂-C₅heterocyclyl; or R^G7and R^G8, R^G8and R^G9, R^G9and R^G10, and/or R^G10and R^G11, together with the carbon atoms to which each is attached, combine to form optionally substituted C₆-C₁₀aryl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heteroaryl, or C₂-C₉heterocyclyl.

In some embodiments, each of R^G7, R^G8, R^G9, R^G10, and R^G11is, independently, H, F, Cl,

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In some embodiments, R^G8is

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In some embodiments, G is

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In some embodiments, R^G7is H; R^G8is

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R^G9is H; and R^G11is H.

In some embodiments, G is

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where

each of R^G12, R^G13, and R^G14is, independently, H, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₂-C₉heteroaryl, optionally substituted C₂-C₆alkenyl, optionally substituted C₂-C₆heteroalkenyl, optionally substituted —O—C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₂-C₅heterocyclyl, hydroxyl, thiol, or optionally substituted amino; or R^G12and R^G14, together with the carbon atoms to which each is attached, combine to form optionally substituted C₆-C₁₀aryl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heteroaryl, or optionally substituted C₂-C₉heterocyclyl.

In some embodiments, each of R^G12, R^G13, and R^G14is, independently, H, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₂-C₉heteroaryl, optionally substituted C₂-C₆alkenyl, optionally substituted C₂-C₆heteroalkenyl, hydroxyl, thiol, or optionally substituted amino; or R^G12and R^G14, together with the carbon atoms to which each is attached, combine to form optionally substituted C₆-C₁₀aryl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heteroaryl, or optionally substituted C₂-C₉heterocyclyl.

In some embodiments, the compound of Formula IA has the structure of Formula IAa:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IA has the structure of Formula IAb:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IA has the structure of Formula IAc:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IA has the structure of Formula IAd:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IA has the structure of Formula IAe:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IA has the structure of Formula IAf:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IA has the structure of Formula IAg:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IA has the structure of Formula IAh:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IB has the structure of Formula IBa:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IB has the structure of Formula IBb:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IB has the structure of Formula IBc:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IB has the structure of Formula IBd:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IB has the structure of Formula IBe:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IB has the structure of Formula IBf:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IB has the structure of Formula IBg:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IB has the structure of Formula IBh:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IC has the structure of Formula ICa:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IC has the structure of Formula ICb:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula ID has the structure of Formula IDa:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula ID has the structure of Formula IDb:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IE has the structure of Formula IEa:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IF has the structure of Formula IF:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound has the structure of any one of compounds B1-B11 in Table 1, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound has the structure of any one of compounds B1-B10 in Table 1, or a pharmaceutically acceptable salt thereof.

In an aspect, the disclosure features a compound having the structure of any one of compounds B1-B11 in Table 1, or a pharmaceutically acceptable salt thereof.

In another aspect, the disclosure features a compound having the structure of any one of compounds B1-B10 in Table 1, or a pharmaceutically acceptable salt thereof.

TABLE 1

Compounds B1-B11 of the Disclosure

Comp-

ound

No.
Structure

B1

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B10

B11

In an aspect, the disclosure features a compound having the structure of Formula II:

A-L-B Formula II,

where

L is a linker;

B is a degradation moiety; and

A has the structure of Formula III:

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where

R¹is H, optionally substituted C₁-C₆alkyl, optionally substituted C₂-C₆alkenyl, optionally substituted C₁-C₆heteroalkyl, or optionally substituted C₃-C₁₀carbocyclyl;

Z¹is CR²or N;

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X¹is CR^X1or N;

X²is O or S;

R^X1is H or optionally substituted C₁-C₆alkyl;

R³″ is H,

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cyano, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₁-C₆alkoxy, optionally substituted amino, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₃-C₁₀heterocyclyl, or optionally substituted C₂-C₉heteroaryl;

R³′ is absent, optionally substituted C₁-C₆alkylene, optionally substituted C₂-C₉heteroarylene, or optionally substituted C₁-C₆heteroalkylene;

G″ is

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optionally substituted C₃-C₁₀carbocyclyl, C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, or optionally substituted C₂-C₉heteroaryl;

G′ is optionally substituted C₃-C₁₀carbocyclylene, C₂-C₉heterocyclylene, optionally substituted C₆-C₁₀arylene, or optionally substituted C₂-C₉heteroarylene; and

A¹is a bond between A and the linker,

where G″ is

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or R³″ is

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or a pharmaceutically acceptable salt thereof.

In some embodiments,

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In some embodiments,

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In some embodiments,

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In some embodiments,

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In some embodiments,

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In some embodiments,

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In some embodiments, the compound of Formula III has the structure of Formula IIIA:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula III has the structure of Formula IIIB:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula III has the structure of Formula IIIC:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula III has the structure of Formula IIIC:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula III has the structure of Formula IIIE:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula III has the structure of Formula IIIF:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, optionally substituted C₁-C₆alkyl is C₁-C₆perfluoroalkyl.

In some embodiments, R¹is

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In some embodiments, R¹is

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In some embodiments, R¹is

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In some embodiments, R¹is H,

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In some embodiments, R¹is

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In some embodiments, R¹is H,

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In some embodiments, R¹is H,

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In some embodiments, R¹is H,

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In some embodiments, R¹is H or

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In some embodiments, R¹is H. In some embodiments, R¹is

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In some embodiments, Z¹is CR². In some embodiments, Z¹is N.

In some embodiments, R²is H, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₃-C₁₀carbocyclyl, or optionally substituted C₆-C₁₀aryl.

In some embodiments, R²is H, halogen, or optionally substituted C₁-C₆alkyl.

In some embodiments, R²is H, F, or

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In some embodiments, R²is H. In some embodiments, R²is F. In some embodiments, R²is

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In some embodiments, R^X1is H. In some embodiments, R^X1is

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In some embodiments, X¹is CH or N. In some embodiments, X¹is CH. In some embodiments, X¹is N.

In some embodiments, X²is O. In some embodiments, X²is S.

In some embodiments, X¹is CH and X²is S.

In some embodiments, R³″ is H, cyano, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₁-C₆alkoxy, optionally substituted amino, optionally substituted C₃-C₁₀heterocyclyl, or optionally substituted C₂-C₉heteroaryl. In some embodiments, R³″ is optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, or optionally substituted C₃-C₁₀heterocyclyl. In some embodiments, R³″ is cyano, optionally substituted C₁-C₆alkoxy, or optionally substituted amino.

In some embodiments, R³″ is H, optionally substituted C₁-C₆alkyl, or optionally substituted C₁-C₆heteroalkyl. In some embodiments, R³″ is H or optionally substituted C₁-C₆alkyl. In some embodiments, R³″ is H. In some embodiments, R³″ is optionally substituted C₁-C₆alkyl.

In some embodiments, R³″ is optionally substituted C₂-C₉heteroaryl or optionally substituted C₁-C₆heteroalkyl. In some embodiments, R³″ is optionally substituted C₁-C₆heteroalkyl.

In some embodiments, R³″ is

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where

W¹is NR⁴or O;

W²is NH, O, or S;

W³is NR′ or O;

R⁴is H or optionally substituted C₁-C₆alkyl;

R⁵is optionally substituted C₁-C₆alkyl, optionally substituted C₃-C₁₀carbocyclyl, or optionally substituted C₂-C₉heterocyclyl;

R⁶is H, optionally substituted C₁-C₆alkyl, optionally substituted C₃-C₁₀carbocyclyl, or optionally substituted C₂-C₉heterocyclyl;

R⁷is H or optionally substituted C₁-C₆alkyl; and

R⁸is H, optionally substituted C₁-C₆alkyl, optionally substituted C₃-C₁₀carbocyclyl, or optionally substituted C₂-C₉heterocyclyl.

In some embodiments, R³″ is optionally substituted C₂-C₉heteroaryl,

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In some embodiments, R³″ is optionally substituted C₂-C₉heteroaryl. In some embodiments, R³″

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In some embodiments, R³″ is

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In some embodiments, R³″ is

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In some embodiments, W¹is NR⁴. In some embodiments, W¹is O.

In some embodiments, W²is NH. In some embodiments, W²is O. In some embodiments, W²is S.

In some embodiments, W³is NR⁷. In some embodiments, W³is O.

In some embodiments, R³″ is

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In some embodiments, R³″ is

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In some embodiments, R³″ is

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In some embodiments, R⁴is H,

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In some embodiments, R⁵is

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In some embodiments, R⁵is

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In some embodiments, R⁶is

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In some embodiments, R⁶is

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In some embodiments, R⁶is H,

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In some embodiments, R⁶is H,

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In some embodiments, R⁷is H,

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In some embodiments, R³″ is H. In some embodiments, R³″ is cyano. In some embodiments, R³″ is

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In some embodiments, R³″ is

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In some embodiments, R³″ is

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In some embodiments, R³″ is

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In some embodiments, R³″ is

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In some embodiments, R³″ is

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In some embodiments, R³″ is

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In some embodiments, G″ is

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In some embodiments, G′ is optionally substituted C₃-C₁₀carbocyclylene or optionally substituted C₂-C₉heterocyclylene. In some embodiments, G′ is optionally substituted C₆-C₁₀arylene or optionally substituted C₂-C₉heteroarylene.

In some embodiments, G′ is optionally substituted C₃-C₁₀carbocyclylene. In some embodiments, G′ is optionally substituted C₆-C₁₀arylene. In some embodiments, G′ is optionally substituted C₂-C₉heterocyclylene. In some embodiments, G′ is optionally substituted C₂-C₉heteroarylene.

In some embodiments, G′ is

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where

each of R^G1′, R^G2′, R^G3′, R^G4′, and R^G6′ is, independently, H, A¹, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₂-C₉heteroaryl, optionally substituted C₂-C₆alkenyl, optionally substituted C₂-C₆heteroalkenyl, optionally substituted —O—C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₂-C₅heterocyclyl, hydroxyl, thiol, or optionally substituted amino; or R^G1′ and R^G2′, R^G2′ and R^G3′, R^G3′ and R^G4′, and/or R^G4′ and R^G5′, together with the carbon atoms to which each is attached, combine to form

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is optionally substituted C₆-C₁₀aryl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heteroaryl, or optionally substituted C₂-C₉heterocyclyl, any of which is optionally substituted with A¹, where one of R^G1′, R^G2′, R^G3′, R^G4′, and R^G5′ is A¹, or

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is substituted with A¹.

In some embodiments, each of R^G1′, R^G2′, R^G3′, R^G4′, and R^G5′ is, independently, H, A¹, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₂-C₉heteroaryl, optionally substituted C₂-C₆alkenyl, optionally substituted C₂-C₆heteroalkenyl, optionally substituted —O—C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₂-C₅heterocyclyl, hydroxyl, thiol, or optionally substituted amino; or R^G1′ and R^G2′, R^G2′ and R^G3′, R^G3′ and R^G4′, and/or R^G4′ and R^G5′, together with the carbon atoms to which each is attached, combine to form

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is substituted with A¹.

In some embodiments, each of R^G1′, R^G2′, R^G3′, R^G4′, and R^G5′ is, independently, H, A¹, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted —O—C₃-C₆carbocyclyl, or optionally substituted —C₁-C₃alkyl-C₂-C₅heterocyclyl; or R^G1′ and R^G2′, R^G2′ and R^G3′, R^G3′ and R^G4′, and/or R^G4′ and R^G5′, together with the carbon atoms to which each is attached, combine to form

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is optionally substituted C₂-C₉heteroaryl or optionally substituted C₂-C₉heterocyclyl, any of which is optionally substituted with A¹, where one of R^G1′, R^G2′, R^G3′, R^G4′, and R^G5′ is A¹, or

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is substituted with A¹.

In some embodiments, each of R^G1′, R^G2′, R^G3′, R^G4′, and R^G5′ is, independently, H, A¹, F, Cl,

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In some embodiments, each of R^G1′, R^G2′, R^G3′, R^G4′, and R^G5′ is, independently, H, A¹, F,

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In some embodiments, each of R^G1′, R^G2′, R^G3′, R^G4′, and R^G5′ is, independently, H, A¹, F, Cl,

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In some embodiments, R^G3′ is A¹.

In some embodiments, R^G1′ is H; R^G2′ is

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R^G3′ is A¹; R^G4′ is

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and R^G5′ is H. In some embodiments, R^G1′ is H; R^G2′ is

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R^G3′ is A¹; R^G4′ is H; and R^G5′ is

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In some embodiments, R^G1′ is H; R^G2′ is

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R^G3′ is A¹; R^G4′ is H; and R^G5′ is H. In some embodiments, R^G1′ is H; R^G2′ is

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R^G3′ is A¹; R^G4′ is H; and R^G5′ is H. In some embodiments, R^G1′ is H; R^G2′ is

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R^G3′ is A¹; R^G4′ is

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and R^G5′ is H.

In some embodiments, R^G1′ and R^G2′, R^G2′ and R^G3′, R^G3′ and R^G4′, and/or R^G4′ and R^G5′, together with the carbon atoms to which each is attached, combine to form

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is optionally substituted C₂-C₉heterocyclyl, which is optionally substituted with A¹, where one of R^G1′, R^G2′, R^G3′, R^G4′, and R^G5′ is A¹, or

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is substituted with A¹. In some embodiments, R^G1′ and R^G2′, R^G2′ and R^G3′, R^G3′ and R^G4′, and/or R^G4′ and R^G5′, together with the carbon atoms to which each is attached, combine to form

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is optionally substituted C₂-C₉heteroaryl, which is optionally substituted with A¹, where one of R^G1′, R^G2′, R^G3′, R^G4′, and R^G5′ is A¹, or

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is substituted with A¹.

In some embodiments, G′ is

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where R^G6′ is H, A¹, or optionally substituted C₁-C₆alkyl. In some embodiments, G′ is

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where R^G6′ is H, A¹, or optionally substituted C₁-C₆alkyl.

In some embodiments, R^G1′ and R^G2′, R^G2′ and R^G3′, R^G3′ and R^G4′, and/or R^G4′ and R^G5′, together with the carbon atoms to which each is attached, combine to form

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is optionally substituted C₂-C₉heterocyclyl or optionally substituted C₂-C₉heteroaryl, any of which is optionally substituted with A¹, where one of R^G1′, R^G2′, R^G3′, R^G4′, and R^G5′ is A¹, or

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is substituted with A¹.

In some embodiments, G′ is

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where R^G6′ is H, A¹, or optionally substituted C₁-C₆alkyl.

In some embodiments, R^G6′ is H, A¹,

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In some embodiments, R^G6′ is H, A¹, or

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In some embodiments, R^G6′ is H or A¹.

In some embodiments, R^G6′ is H. In some embodiments, R^G6′ is A¹.

In some embodiments, R^G1′ is H, A¹, F,

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In some embodiments, RG″ is H.

In some embodiments, R^G2′ is H, A¹, F,

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In some embodiments, R^G2′ is H.

In some embodiments, R^G3′ is H, A¹, F,

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In some embodiments, R^G3′ is H.

In some embodiments, R^G4′ is H, A¹, F,

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In some embodiments, R^G4′ is H.

In some embodiments, R^G5′ is H, A¹, F,

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In some embodiments, R^G5′ is H.

In some embodiments, one or more of R^G1′, R^G2′, R^G3′, R^G4′, and R^G5′ is H. In some embodiments, two or more of R^G1′, R^G2′, R^G3′, R^G4′, and R^G5′ is H. In some embodiments, three or more of R^G1′, R^G2′, R^G3′, R^G4′, and R^G5′ is H.

In some embodiments, R^G1′ is A¹. In some embodiments, R^G2′ is A¹. In some embodiments, R^G3′ is A¹. In some embodiments, R^G4′ is A¹. In some embodiments, R^G5′ is A¹. In some embodiments,

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is substituted with A¹.

In some embodiments, G′ is

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where

each of R^G7′, R^G8′, R^G9′, R^G10′, and R^G11′ is, independently, H, A¹, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₂-C₉heteroaryl, optionally substituted C₂-C₆alkenyl, optionally substituted C₂-C₆heteroalkenyl, optionally substituted —O—C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₂-C₅heterocyclyl, hydroxyl, thiol, or optionally substituted amino; or R^G7′ and R^G8′, R^G8′ and R^G9′, R^G9′ and R^G10′, and/or R^G10′ and R^G11′, together with the carbon atoms to which each is attached, combine to form

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is optionally substituted C₆-C₁₀aryl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heteroaryl, or C₂-C₉heterocyclyl, any of which is optionally substituted with A¹, where one of R^G7′, R^G8′, R^G9′, R^G10′, and R^G11′ is A¹; or

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is substituted with A¹.

In some embodiments, each of R^G7′, R^G8′, R^G9′, R^G10′, and R^G11′ is, independently, H, A¹, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₂-C₉heteroaryl, optionally substituted C₂-C₆alkenyl, optionally substituted C₂-C₆heteroalkenyl, optionally substituted —O—C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₂-C₅heterocyclyl, hydroxyl, thiol, or optionally substituted amino; or R^G7′ and R^G8′, R^G8′ and R^G9′, R^G9′ and R^G10′, and/or R^G10′ and R^G11′, together with the carbon atoms to which each is attached, combine to form

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is substituted with A¹.

In some embodiments, each of R^G7′, R^G8′, R^G9′, R^G10′, and R^G11′ is, independently, H, A¹, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted —O—C₃-C₆carbocyclyl, or optionally substituted —C₁-C₃alkyl-C₂-C₅heterocyclyl; or R^G7′ and R^G8′, R^G8′ and R^G9′, R^G9′ and R^G10′, and/or R^G10′ and R^G11′, together with the carbon atoms to which each is attached, combine to form

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is substituted with A¹.

In some embodiments, each of R^G7′, R^G8′, R^G9′, R^G10′, and R^G11′ is, independently, H, A¹, F, Cl,

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In some embodiments, R^G8′ is

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In some embodiments, G′ is

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In some embodiments, R^G7′ is H; R^G8′ is

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R^G9′ is A¹; and R^G11′ is H.

In some embodiments, G′ is

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where

each of R^G12′, R^G13′, and R^G14′ is, independently, H, A¹, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₂-C₉heteroaryl, optionally substituted C₂-C₆alkenyl, optionally substituted C₂-C₆heteroalkenyl, optionally substituted —O—C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₂-C₅heterocyclyl, hydroxyl, thiol, or optionally substituted amino; or R^G12′ and R^G14′, together with the carbon atoms to which each is attached, combine to form

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is substituted with A¹.

In some embodiments, each of R^G12′, R^G13′, and R^G14′ is, independently, H, A¹, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₂-C₉heteroaryl, optionally substituted C₂-C₆alkenyl, optionally substituted C₂-C₆heteroalkenyl, optionally substituted —O—C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₂-C₅heterocyclyl, hydroxyl, thiol, or optionally substituted amino; or R^G12′ and R^G14′, together with the carbon atoms to which each is attached, combine to form

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is substituted with A¹.

In some embodiments, R³″ is

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In some embodiments, R³′ is absent.

In some embodiments, R³″ is

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and R³′ is absent.

In some embodiments, R³′ is optionally substituted C₁-C₆alkylene, optionally substituted C₂-C₉heteroarylene, or optionally substituted C₁-C₆heteroalkylene. In some embodiments, optionally substituted C₂-C₉heteroarylene or optionally substituted C₁-C₆heteroalkylene.

In some embodiments, R³′ is

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where

W¹is NR⁴′ or O;

W²is NH, O, or S;

W³is NR⁷′ or O;

R⁴′ is H or optionally substituted C₁-C₆alkyl;

R⁵′ is absent, optionally substituted C₁-C₆alkylene, optionally substituted C₃-C₁₀carbocyclylene, or optionally substituted C₂-C₉heterocyclylene;

R⁶′ is absent, optionally substituted C₁-C₆alkylene, optionally substituted C₃-C₁₀carbocyclylene, or optionally substituted C₂-C₉heterocyclylene,

R⁷′ is H or optionally substituted C₁-C₆alkyl; and

R⁸′ is absent, optionally substituted C₁-C₆alkyl, optionally substituted C₃-C₁₀carbocyclyl, or optionally substituted C₂-C₉heterocyclyl.

In some embodiments, R³′ is optionally substituted C₂-C₉heteroarylene,

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In some embodiments, R³′ is optionally substituted C₂-C₉heteroarylene. In some embodiments, R³′ is

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In some embodiments, R³′ is

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In some embodiments, W¹is NR^4′. In some embodiments, W¹is O.

In some embodiments, W²is NH. In some embodiments, W²is O. In some embodiments, W²is S.

In some embodiments, W³is NR″. In some embodiments, W³is O.

In some embodiments, R³′ is

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In some embodiments, R³′ is

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In some embodiments, R³′ is

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In some embodiments, R⁴′ is H,

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In some embodiments, R⁷′ is H,

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In some embodiments, G″ is optionally substituted C₃-C₁₀carbocyclyl or optionally substituted C₂-C₉heterocyclyl. In some embodiments, G″ is optionally substituted C₆-C₁₀aryl or optionally substituted C₂-C₉heteroaryl.

In some embodiments, G″ is optionally substituted C₃-C₁₀carbocyclyl. In some embodiments, G is optionally substituted C₆-C₁₀aryl. In some embodiments, G is optionally substituted C₂-C₉heterocyclyl. In some embodiments, G″ is optionally substituted C₂-C₉heteroaryl.

In some embodiments, G″ is

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where

each of R^G1, R^G2, R^G3, R^G4, and R^G5is, independently, H, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₂-C₉heteroaryl, optionally substituted C₂-C₆alkenyl, optionally substituted C₂-C₆heteroalkenyl, optionally substituted —O—C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₃-C₆carbocyclyl, optionally substituted —C₁-C₃alkyl-C₂-C₅heterocyclyl, hydroxyl, thiol, or optionally substituted amino; or R^G1and R^G2, R^G2and R^G3, R^G3and R^G4, and/or R^G4and R^G5, together with the carbon atoms to which each is attached, combine to form optionally substituted C₆-C₁₀aryl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heteroaryl, or optionally substituted C₂-C₉heterocyclyl.

In some embodiments, each of R^G1, R^G2, R^G3, R^G4, and R^G5is, independently, H, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted —O—C₃-C₆carbocyclyl, or optionally substituted —C₁-C₃alkyl-C₂-C₅heterocyclyl; or R^G1and R^G2, R^G2and R^G3, R^G3and R^G4, and/or R^G4and R^G5, together with the carbon atoms to which each is attached, combine to form optionally substituted C₂-C₉heteroaryl or optionally substituted C₂-C₉heterocyclyl.

In some embodiments, each of R^G1, R^G2, R^G3, R^G4, and R^G5is, independently, H, F, Cl,

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In some embodiments, each of R^G1, R^G2, R^G3, R^G4, and R^G5is, independently, H, F,

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In some embodiments, each of R^G1, R^G2, R^G3, R^G4, and R^G5is, independently, H, F, Cl

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In some embodiments, R^G1is H; R^G2is

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R^G3is

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R^G4is

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and R^G5is H. In some embodiments, R^G1is H; R^G2is

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R^G3is

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R^G4is H; and R^G5is

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In some embodiments, R^G1is H; R^G2is

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R^G3is

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R^G4is Cl or F; and R^G5is H. In some embodiments, R^G1is H; R^G2is

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R^G3is

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R^G4is H; and R^G5is H. In some embodiments, R^G1is H; R^G2is

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R^G3is

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R^G4is

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and R^G5is H.

In some embodiments, R^G1and R^G2, R^G2and R^G3, R^G3and R^G4, and/or R^G4and R^G5, together with the carbon atoms to which each is attached, combine to form optionally substituted C₂-C₉heterocyclyl. In some embodiments, R^G1and R^G2, R^G2and R^G3, R^G3and R^G4, and/or R^G4and R^G6, together with the carbon atoms to which each is attached, combine to form optionally substituted C₂-C₉heteroaryl.

In some embodiments, R^G1and R^G2, R^G2and R^G3, R^G3and R^G4, and/or R^G4and R^G6, together with the carbon atoms to which each is attached, combine to form optionally substituted C₂-C₉heterocyclyl. In some embodiments, R^G1and R^G2, R^G2and R^G3, R^G3and R^G4, and/or R^G4and R^G6, together with the carbon atoms to which each is attached, combine to form optionally substituted C₂-C₉heteroaryl.

In some embodiments, G″ is

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where R^G6is H or optionally substituted C₁-C₆alkyl. In some embodiments, G″ is

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where R^G6is H or optionally substituted C₁-C₆alkyl.

In some embodiments, G″ is

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where R^G6is H or optionally substituted C₁-C₆alkyl.

In some embodiments, R^G6is H,

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In some embodiments, R^G6is H or

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In some embodiments, R^G6is H.

In some embodiments, R^G1is H, F,

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In some embodiments, R^G1is H.

In some embodiments, R^G2is H, F,

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In some embodiments, R^G2is H.

In some embodiments, R^G3is H, F,

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In some embodiments, R^G3is H.

In some embodiments, R^G4is H, F,

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In some embodiments, R^G4is H.

In some embodiments, R^G5is H, F,

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In some embodiments, R^G5is H.

In some embodiments, G″ is

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where

In some embodiments, each of R^G7, R^G8, R^G9, R^G10, and R^G11is, independently, H, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted —O—C₃-C₆carbocyclyl, or optionally substituted —C₁-C₃alkyl-C₂-C₅heterocyclyl; or R^G7and R^G8, R^G8and R^G9, R^G9and R^G10, and/or R^G10and R^G11, together with the carbon atoms to which each is attached, combine to form optionally substituted C₆-C₁₀aryl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heteroaryl, or C₂-C₉heterocyclyl.

In some embodiments, each of R^G7, R^G8, R^G9, R^G10, and R^G11is, independently, H, F, Cl,

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In some embodiments, R^G8is

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In some embodiments, G″ is

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In some embodiments, R^G7is H; R^G8is

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R^G9is H; and R^G11is H.

In some embodiments, G″ is

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where

In some embodiments, A has the structure of Formula IIIAa:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, A has the structure of Formula IIIAb:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, A has the structure of Formula IIIAc:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, A has the structure of Formula IIIAd:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, A has the structure of Formula IIIAe:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, A has the structure of Formula IIIAf:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, A has the structure of Formula IIIAg:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, A has the structure of Formula lllAh:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, A has the structure of Formula IIIBa:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, A has the structure of Formula IIIBb:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, A has the structure of Formula IIIBc:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, A has the structure of Formula IIIBd:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, A has the structure of Formula IIIBe:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, A has the structure of Formula IIIBf:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, A has the structure of Formula IIIBg:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, A has the structure of Formula IIIBh:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IIIC has the structure of Formula IIICa:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IIIC has the structure of Formula IIICb:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IIID has the structure of Formula IIIDa:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IIID has the structure of Formula IIIDb:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IIIE has the structure of Formula IIIEa:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula IIIF has the structure of Formula IIIF:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the degradation moiety is a ubiquitin ligase binding moiety.

In some embodiments, the ubiquitin ligase binding moiety comprises Cereblon ligands, IAP (Inhibitors of Apoptosis) ligands, mouse double minute 2 homolog (MDM2), or von Hippel-Lindau (VHL) ligands, or derivatives or analogs thereof.

In some embodiments, the degradation moiety is a ubiquitin ligase binding moiety.

In some embodiments, the degradation moiety includes the structure of Formula Y:

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where

A²is a bond between the degradation moiety and the linker;

v1 is 0, 1, 2, 3, 4, or 5;

u1 is 1, 2, or 3;

T¹is a bond or

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T²is

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R^5Ais H, optionally substituted C₁-C₆alkyl, or optionally substituted C₁-C₆heteroalkyl;

each R^J1is, independently, halogen, optionally substituted C₁-C₆alkyl, or optionally substituted C₁-C₆heteroalkyl;

J^Ais absent, O, optionally substituted amino, optionally substituted C₁-C₆alkyl, or optionally substituted C₁-C₆heteroalkyl; and

J is absent, optionally substituted C₃-C₁₀carbocyclylene, optionally substituted C₆-C₁₀arylene, optionally substituted C₂-C₉heterocyclylene, or optionally substituted C₂-C₉heteroarylene, or a pharmaceutically acceptable salt thereof.

In some embodiments, T²is

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In some embodiments, T²is

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In some embodiments, T²is

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In some embodiments, T²is

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In some embodiments, the structure of Formula Y has the structure of Formula Y1:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, T¹is a bond. In some embodiments, T¹is

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In some embodiments, the structure of Formula Y has the structure of Formula Y2:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the structure of Formula Y has the structure of Formula Z:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, u1 is 1. In some embodiments, u1 is 2. In some embodiments u1 is 3.

In some embodiments, the structure of Formula Z has the structure of Formula AA0:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the structure of Formula Z has the structure of Formula AB:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the structure of Formula Z has the structure of Formula AC:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, J^Ais absent. In some embodiments, J^Ais optionally substituted C₁-C₆alkyl. In some embodiments, J^Ais optionally substituted C₁-C₆heteroalkyl. In some embodiments, J^Ais O or optionally substituted amino.

In some embodiments, J^Ais

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In some embodiments, the structure of Formula AA0 has the structure of Formula AA0:

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or a pharmaceutically acceptable salt thereof.

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

In some embodiments, the structure of Formula AA has the structure of Formula AA1:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the structure of Formula AB has the structure of Formula AB1:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the structure of Formula AC has the structure of Formula AC1:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, J is absent. In some embodiments, J is optionally substituted C₃-C₁₀carbocyclylene or optionally substituted C₆-C₁₀arylene. In some embodiments, J is optionally substituted C₂-C₉heterocyclylene or optionally substituted C₂-C₉heteroarylene.

In some embodiments, J is optionally substituted heterocyclylene. In some embodiments, J is optionally substituted C₆-C₁₀arylene.

In some embodiments, J is

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In some embodiments, the structure of Formula AA has the structure of Formula AA2:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the structure of Formula AA has the structure of Formula AA3:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the structure of Formula AA has the structure of Formula AA4:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, R^A5is H or optionally substituted C₁-C₆alkyl. In some embodiments, R^A5is optionally substituted C₁-C₆heteroalkyl.

In some embodiments, R^A5is H or methyl. In some embodiments, R^A5is H. In some embodiments, R^A5is methyl. In some embodiments, R^A5is

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In some embodiments, the structure of Formula AA has the structure of Formula A:

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where

Y¹is

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R^A5is H, optionally substituted C₁-C₆alkyl, or optionally substituted C₁-C₆heteroalkyl;

R^A6is H or optionally substituted C₁-C₆alkyl; and R^A7is H or optionally substituted C₁-C₆alkyl; or R^A6and R^A7, together with the carbon atom to which each is bound, combine to form optionally substituted C₃-C₆carbocyclyl or optionally substituted C₂-C₅heterocyclyl; or R^A6and R^A7, together with the carbon atom to which each is bound, combine to form optionally substituted C₃-C₆carbocyclyl or optionally substituted C₂-C₅heterocyclyl;

R^A8is H, optionally substituted C₁-C₆alkyl, or optionally substituted C₁-C₆heteroalkyl;

each of R^A1, R^A2, R^A3, and R^A4is, independently, H, A², halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₂-C₉heteroaryl, optionally substituted C₂-C₆alkenyl, optionally substituted C₂-C₆heteroalkenyl, optionally substituted —O—C₃-C₆carbocyclyl, hydroxyl, thiol, or optionally substituted amino; or R^A1and R^A2, R^A2and R^A3, and/or R^A3and R^A4, together with the carbon atoms to which each is attached, combine to form

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is optionally substituted C₆-C₁₀aryl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heteroaryl, or C₂-C₉heterocyclyl, any of which is optionally substituted with A², where one of R^A1, R^A2, R^A3, and R^A4is A², or

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is substituted with A², or a pharmaceutically acceptable salt thereof.

In some embodiments, each of R^A1, R^A2, R^A3, and R^A4is, independently, H, A², halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₂-C₉heteroaryl, optionally substituted C₂-C₆alkenyl, optionally substituted C₂-C₆heteroalkenyl, hydroxyl, thiol, or optionally substituted amino; or R^A1and R^A2, R^A2and R^A3, and/or R^A3and R^A4, together with the carbon atoms to which each is attached, combine to form

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is optionally substituted C₆-C₁₀aryl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heteroaryl, or C₂-C₉heterocyclyl, any of which is optionally substituted with A², where one of R^A1, R^A2, R^A3, and R^A4is A², or

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is substituted with A², or a pharmaceutically acceptable salt thereof.

In some embodiments, each of R^A1, R^A2, R^A3, and R^A4is, H, A², halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted —O—C₃-C₆carbocyclyl, hydroxyl, optionally substituted amino; or R^A1and R^A2, R^A2and R^A3, or R^A3and R^A4, together with the carbon atoms to which each is attached, combine to form

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is optionally substituted C₂-C₉heterocyclyl, which is optionally substituted with A², where one of R^A1, R^A2, R^A3, and R^A4is A², or

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is substituted with A².

In some embodiments, each of R^A1, R^A2, R^A3, and R^A4is, independently, H, A², F,

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or R^A1and R^A2, R^A2and R^A3, or R^A3and R^A4, together with the carbon atoms to which each is attached, combine to form

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is optionally substituted C₂-C₉heterocyclyl, which is optionally substituted with A², where one of R^A1, R^A2, R^A3, and R^A4is A², or

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is substituted with A².

In some embodiments, R^A1is A². In some embodiments, R^A2is A². In some embodiments, R^A3is A². In some embodiments, R^A4is A². In some embodiments, R^A5is A².

In some embodiments, R^A5is H or optionally substituted C₁-C₆alkyl.

In some embodiments, R^A5is H or

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In some embodiments, R^A5is H. In some embodiments, R^A5is

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In some embodiments, Y¹is

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In some embodiments, Y¹is

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In some embodiments, Y¹is

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In some embodiments, each of R^A6and R^A7is, independently, H, F,

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or R^A6and R^A7, together with the carbon atom to which each is bound, combine to form

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In some embodiments, R^A6is H and R^A7is H.

In some embodiments, Y¹is

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In some embodiments, Y¹is

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In some embodiments, Y¹is

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

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or a pharmaceutically acceptable salt thereof.

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

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or a pharmaceutically acceptable salt thereof.

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

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the structure of Formula A has the structure of Formula A4:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the structure of Formula A has the structure of Formula A5:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the structure of Formula A has the structure of Formula A6:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the structure of Formula A has the structure of Formula A7:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the structure of Formula A has the structure of Formula A8:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the structure of Formula A has the structure of Formula A9:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the structure of Formula A has the structure of Formula A10:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, wherein the structure of Formula A is

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or derivative or analog thereof.

In some embodiments, the structure of Formula A is

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In some embodiments, the structure of Formula A is

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or derivative or analog thereof.

In some embodiments,

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where R^A9is H, A², optionally substituted C₁-C₆alkyl, or optionally substituted C₁-C₆heteroalkyl.

In some embodiments, the structure of Formula A is

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In some embodiments, R^A9is H, A², or optionally substituted C₁-C₆alkyl. In some embodiments, R^A9is H, A², or methyl. In some embodiments, R^9Ais H. In some embodiments, R^9Ais methyl. In some embodiments, R^A9is A².

In some embodiments, the structure of Formula A is

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In some embodiments, the structure of Formula AA has the structure of Formula B:

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where

R^A5is H, optionally substituted C₁-C₆alkyl, or optionally substituted C₁-C₆heteroalkyl;

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is optionally substituted C₆-C₁₀aryl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heteroaryl, or C₂-C₉heterocyclyl, any of which is optionally substituted with A², where one of R^A1, R^A2, R^A3, and R^A4is A², or

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is substituted with A², or a pharmaceutically acceptable salt thereof.

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is optionally substituted C₂-C₉heterocyclyl, which is optionally substituted with A², where one of R^A1, R^A2, R^A3, and R^A4is A², or

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is substituted with A².

In some embodiments, each of R^A1, R^A2, R^A3, and R^A4is, independently, H, A², F,

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or R^A1and R^A2, R^A2and R^A3, or R^A3and R^A4, together with the carbon atoms to which each is attached, combine to form

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is optionally substituted C₂-C₉heterocyclyl, which is optionally substituted with A², where one of R^A1, R^A2, R^A3, and R^A4is A², or

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is substituted with A².

In some embodiments, R^A1is A². In some embodiments, R^A2is A². In some embodiments, R^A3is A². In some embodiments, R^A4is A2. In some embodiments, R^A5is A².

In some embodiments, R^A5is H or optionally substituted C₁-C₆alkyl.

In some embodiments, R^A5is H or

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In some embodiments, R^A5is H. In some embodiments, R^A5is

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In some embodiments, the structure of Formula B has the structure of Formula B1:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the structure of Formula B has the structure of Formula B2:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the structure of Formula B has the structure of Formula B3:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the structure of Formula B has the structure of Formula B4:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the structure of Formula B is

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In some embodiments, the structure of Formula B is

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In some embodiments, the structure of Formula B is

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

In some embodiments, the von Hippel-Lindau ligand has the structure of

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or derivative or analog thereof.

In some embodiments, the degradation moiety includes the structure of Formula C:

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where

R^B1is H, A², optionally substituted C₁-C₆alkyl, or optionally substituted C₁-C₆heteroalkyl;

R^B2is H, optionally substituted C₁-C₆alkyl, or optionally substituted C₁-C₆heteroalkyl;

R^B3is A², optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₁-C₆alkyl C₃-C₁₀carbocyclyl, or optionally substituted C₁-C₆alkyl C₆-C₁₀aryl;

R^B4is H, optionally substituted C₁-C₆alkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₁-C₆alkyl C₃-C₁₀carbocyclyl, or optionally substituted C₁-C₆alkyl C₆-C₁₀aryl;

R^B5is H, optionally substituted C₁-C₆alkyl, or optionally substituted C₁-C₆heteroalkyl;

v2 is 0, 1, 2, 3, or 4;

each R^B6is, independently, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₂-C₉heteroaryl, optionally substituted C₂-C₆alkenyl, optionally substituted C₂-C₆heteroalkenyl, hydroxy, thiol, or optionally substituted amino; and

each of R^B7and R^B8is, independently, H, halogen, optionally substituted C₁-C₆alkyl, or optionally substituted C₆-C₁₀aryl,

where one of R^B1and R^B3is A², or a pharmaceutically acceptable salt thereof.

In some embodiments, the structure of Formula C is

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or derivative or analog thereof.

In some embodiments, the structure of Formula C is

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In some embodiments, the degrader moiety includes the structure of Formula D:

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where

A²is a bond between B and the linker;

each of R^C1, R^C2, and R^C7is, independently, H, optionally substituted C₁-C₆alkyl, or optionally substituted C₁-C₆heteroalkyl;

R^C3is optionally substituted C₁-C₆alkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₁-C₆alkyl C₃-C₁₀carbocyclyl, or optionally substituted C₁-C₆alkyl C₆-C₁₀aryl;

R^C5is optionally substituted C₁-C₆alkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₁-C₆alkyl C₃-C₁₀carbocyclyl, or optionally substituted C₁-C₆alkyl C₆-C₁₀aryl;

v3 is 0, 1, 2, 3, or 4;

each R^C8is, independently, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₂-C₉heteroaryl, optionally substituted C₂-C₆alkenyl, optionally substituted C₂-C₆heteroalkenyl, hydroxy, thiol, or optionally substituted amino;

v4 is 0, 1, 2, 3, or 4; and

each R^C9is, independently, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₂-C₉heteroaryl, optionally substituted C₂-C₆alkenyl, optionally substituted C₂-C₆heteroalkenyl, hydroxy, thiol, or optionally substituted amino, or a pharmaceutically acceptable salt thereof.

In some embodiments, the structure of Formula D is

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or derivative or analog thereof.

In some embodiments, the degrader moiety includes the structure of Formula E:

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where

A²is a bond between B and the linker;

each of R^C10and R^C11is, independently, H, optionally substituted C₁-C₆alkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₁-C₆alkyl C₃-C₁₀carbocyclyl, or optionally substituted C₁-C₆alkyl C₆-C₁₀aryl;

v5 is 0, 1, 2, 3, or 4;

each R^C12is, independently, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₂-C₉heteroaryl, optionally substituted C₂-C₆alkenyl, optionally substituted C₂-C₆heteroalkenyl, hydroxy, thiol, or optionally substituted amino;

v6 is 0, 1, 2, 3, or 4; and

each R²¹is, independently, halogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted C₃-C₁₀carbocyclyl, optionally substituted C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, optionally substituted C₂-C₉heteroaryl, optionally substituted C₂-C₆alkenyl, optionally substituted C₂-C₆heteroalkenyl, hydroxy, thiol, or optionally substituted amino, or a pharmaceutically acceptable salt thereof.

In some embodiments, the structure of Formula E is

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or derivative or analog thereof.

In some embodiments, the degradation moiety includes the structure of Formula FA:

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where

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or a bicyclic moiety which is substituted with A²and substituted with one or more groups independently selected from H, R^FF1, and oxo;

custom-character is a single bond or a double bond;

u2 is 0, 1, 2, or 3;

A²is a bond between the degrader and the linker;

Y^Fais CR^FbR^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;

Y^Fbis NH, NR^FF1, CH₂, CHR^FF1, C(R^FF1)₂, O, or S;

Y^Fcis CR^FdR^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;

each of R^Fb, R^Fc, R^Fd, and R^Feis, independently, H, alkyl, aliphatic, heteroaliphatic, aryl, heteroaryl, carbocyclyl, hydroxyl, alkoxy, amino, —NHalkyl, or —Nalkyl₂;

or R^Fband R^Fc, together with the carbon atom to which each is attached, combine to 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;

or R^Fdand R^Fe, together with the carbon atom to which each is attached, combine to 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

or R^Fdand R^Fb, together with the carbon atoms to which each is attached, combine to form a 1, 2, 3, or 4 carbon bridged ring;

each of Y^Fdand Y^Ffis, independently, CH₂, CHR^FF2, C(R^FF2)₂, C(O), N, NH, NR^FF3, O, S, or S(O);

Y^Feis a bond or a divalent moiety attached to Y^Fdand Y^Ffthat contains 1 to 5 contiguous carbon atoms that form a 3 to 8-membered ring,

- wherein 1, 2, or 3 carbon atoms can be replaced with a nitrogen, oxygen, or sulfur atom;
- wherein one of the ring atoms is substituted with A²and the others are substituted with one or more groups independently selected from H and R^FF1; and
- wherein the contiguous atoms of Y^Fecan be attached through a single or double bond;

each R^FF1is, independently, H, alkyl, alkenyl, alkynyl, aliphatic, heteroaliphatic, carbocyclyl, halogen, hydroxyl, amino, cyano, alkoxy, aryl, heteroaryl, heterocyclyl, alkylamino, alkylhydroxyl, or haloalkyl;

each R^FF2is, independently, alkyl, alkene, alkyne, halogen, hydroxyl, alkoxy, azide, amino, —C(O)H, —C(O)OH, —C(O)(aliphatic, including alkyl), —C(O)O(aliphatic, including alkyl), —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

R^FF3is alkyl, alkenyl, alkynyl, —C(O)H, —C(O)OH, —C(O)alkyl, or —C(O)Oalkyl,

wherein if Y^Fdor Y^Ffis substituted with A², then Y^Feis a bond, or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula FA has the structure of Formula FA1:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the degradation moiety includes the structure of Formula FB:

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where

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or a bicyclic moiety which is substituted with A²and substituted with one or more groups independently selected from H, R^FF1, and oxo;

A²is a bond between the degrader and the linker;

Y^Fais CR^FbR^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;

each of Y^Fband Y^Fgis, independently, NH, NR^FF1, CH₂, CHR^FF1, C(R^FF1)₂, O, or S;

Y^Fcis CR^FdR^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;

each of R^Fb, R^Fc, R^Fd, R^Fe, R^Ff, and R^Fgis independently, H, alkyl, aliphatic, heteroaliphatic, aryl, heteroaryl, carbocyclyl, hydroxyl, alkoxy, amino, —NHalkyl, or —Nalkyl₂;

or R^Ffand R^Fg, together with the carbon atom to which each is attached, combine to 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;

or R^Fdand R^Fb, together with the carbon atoms to which each is attached, combine to form a 1, 2, 3, or 4 carbon bridged ring;

or R^Fdand R^Ff, together with the carbon atoms to which each is attached, combine to form a 1, 2, 3, or 4 carbon bridged ring;

or R^Fband R^Fg, together with the carbon atoms to which each is attached, combine to form a 1, 2, 3, or 4 carbon bridged ring;

each of Y^Fdand Y^Ffis, independently, CH₂, CHR^FF2, C(R^FF2)₂, C(O), N, NH, NR^FF3, O, S, or S(O);

Y^Feis a bond or a divalent moiety attached to Y^Fdand Y^Ffthat contains 1 to 5 contiguous carbon atoms that form a 3 to 8-membered ring,

- wherein 1, 2, or 3 carbon atoms can be replaced with a nitrogen, oxygen, or sulfur atom;
- wherein one of the ring atoms is substituted with A²and the others are substituted with one or more groups independently selected from H and R^FF1; and
- wherein the contiguous atoms of Y^Fecan be attached through a single or double bond;

—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

R^FF3is alkyl, alkenyl, alkynyl, —C(O)H, —C(O)OH, —C(O)alkyl, or —C(O)Oalkyl,

wherein if Y^Fdor Y^Ffis substituted with A², then Y^Feis a bond, or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula FB has the structure of Formula FB1:

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or a pharmaceutically acceptable salt thereof.

In some embodiments, the degradation moiety includes the structure of Formula F1:

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where A²is a bond between the degrader and the linker; and R^F1is absent or O, or a pharmaceutically acceptable salt thereof.

In some embodiments, R^F1is absent. In some embodiments, R^F1is O.

In some embodiments, the structure of Formula F1 is

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In some embodiments, the degradation moiety includes the structure Formula F2:

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where A²is a bond between the degrader and the linker; and Y²is CH₂or NH, or a pharmaceutically acceptable salt thereof.

In some embodiments, Y²is NH. In some embodiments, Y²is CH₂.

In some embodiments, structure of Formula F2 is

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In some embodiments, the degradation moiety includes the structure Formula G:

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where A²is a bond between the degrader and the linker; and Y³is CH₂or NH, or a pharmaceutically acceptable salt thereof.

In some embodiments, Y³is NH. In some embodiments, Y³is CH₂.

In some embodiments, structure of Formula G is

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The degradation moiety may also include structures found in, e.g., WO2017/197036; WO2019/204354, WO2019/236483, WO2020/010177; and WO2020/010227, the structures of which are herein incorporated by reference.

In some embodiments, the linker has the structure of Formula IV:

A¹-(B¹)_f—(C¹)_g—(B²)_h-(D)-(B³)_i—(C²)_j—(B⁴)_k-A² Formula IV

where

A¹is a bond between the linker and A;

A²is a bond between B and the linker;

each of B¹, B², B³, and B⁴is, independently, optionally substituted C₁-C₂alkylene, optionally substituted C₁-C₃heteroalkylene, O, S, S(O)₂, or NR^N;

each R^Nis, independently, H, optionally substituted C_1-4alkyl, optionally substituted C_2-4alkenyl, optionally substituted C_2-4alkynyl, optionally substituted C_2-6heterocyclyl, optionally substituted C_6-12aryl, or optionally substituted C_1-7heteroalkyl;

each of C¹and C²is, independently, carbonyl, thiocarbonyl, sulphonyl, or phosphoryl;

each of f, g, h, i, j, and k is, independently, 0 or 1; and

D is optionally substituted C_1-10alkylene, optionally substituted C_2-10alkenylene, optionally substituted C_2-10alkynylene, optionally substituted C_2-6heterocyclylene, optionally substituted C_6-12arylene, optionally substituted C₂-C₁₀polyethylene glycol, or optionally substituted C_1-10heteroalkylene, or a chemical bond linking A¹-(B¹)_f—(C¹)_g—(B²)_h— to —(B³)_i—(C²)_k—(B⁴)_k-A².

In some embodiments, each of B¹, B², B³, and B⁴is, independently, optionally substituted C₁-C₄alkylene, optionally substituted C₁-C₄heteroalkylene, or NR^N.

In some embodiments, each R^Nis, independently, H or optionally substituted C₁-C₄alkylene.

In some embodiments, each R^Nis, independently, H or methyl.

In some embodiments, each of B¹and B⁴is, independently,

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In some embodiments, B¹is

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In some embodiments, each of C¹and C²is, independently,

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In some embodiments, C¹is

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In some embodiments, B²is NR^N. In some embodiments, B²is optionally substituted C₁-C₄alkylene.

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

In some embodiments, the linker has the structure of

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wherein

x is 1, 2, 3, 4, 5, 6, 7, or 8;

y is 1, 2, 3, or 4;

R^xis H, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, or optionally substituted C₃-C₆carbocyclyl;

R^yis H, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, or optionally substituted C₃-C₆carbocyclyl; and

W is O or NR^w, wherein R^wis H, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, or optionally substituted C₃-C₆carbocyclyl.

In some embodiments, the linker has the structure of

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In some embodiments, R^xis H or me optionally substituted C₁-C₆alkyl. In some embodiments, R^yis H or optionally substituted C₁-C₆alkyl. In some embodiments, R^wis H or optionally substituted C₁-C₆alkyl.

In some embodiments, R^xis H or methyl. In some embodiments, R^yis H or methyl. In some embodiments, R^wis H or methyl.

In some embodiments, the linker has the structure of

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In some embodiments, the linker has the structure of

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In some embodiments, the linker has the structure of

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In some embodiments, the linker has the structure of Formula V:

A¹(E¹)-(F¹)—(C³)_m-(E³)_n-(F²)_o1—(F³)_o2-(E²)_p-A², Formula V

where

A¹is a bond between the linker and A;

A²is a bond between B and the linker;

each of m, n, o1, o2, and p is, independently, 0 or 1;

each of E¹and E²is, independently, O, S, NR^N, optionally substituted C_1-10alkylene, optionally substituted C_2-10alkenylene, optionally substituted C_2-10alkynylene, optionally substituted C₂-C₁₀polyethylene glycol, or optionally substituted C_1-10heteroalkylene;

E³is optionally substituted C₁-C₆alkylene, optionally substituted C₁-C₆heteroalkylene, O, S, or NR^N;

C³is carbonyl, thiocarbonyl, sulphonyl, or phosphoryl; and

each of F¹, F², and F³is, independently, optionally substituted C₃-C₁₀carbocyclylene, optionally substituted C_2-10heterocyclylene, optionally substituted C₆-C₁₀arylene, or optionally substituted C₂-C₉heteroarylene.

In some embodiments, the linker has the structure of Formula Va:

A¹-(E¹)-(F¹)—(C³)_m-(E²)_p-A². Formula Va

In some embodiments, the linker has the structure of Formula Vb:

A¹-(E¹)-(F¹)-(E²)_p-A². Formula Vb

In some embodiments, the linker has the structure of Formula Vc:

A¹-(E¹)-(F¹)-A². Formula Vc

In some embodiments, the linker has the structure of Formula Vd:

A¹-(E¹)-(F¹)—(C³)_m—(F²)_o1-A². Formula Vd

In some embodiments, the linker has the structure of Formula Ve:

A¹-(E¹)-(F¹)-(E³)_n-(F²)_o1-(E²)_p-A². Formula Ve

In some embodiments, the linker has the structure of Formula Vf:

A¹-(E¹)-(F¹)—(C³)_m-(E³)_n-(F₂)_o1-(E²)_p-A². Formula Vf

In some embodiments, the linker has the structure of Formula Vg:

A¹-(E¹)-(F¹)-(E³)_n-(F²)_o1-A², Formula Vg

In some embodiments, each of E and E²is, independently, NR^N, optionally substituted C_1-10alkylene, optionally substituted C₂-C₁₀polyethylene glycolene, or optionally substituted C_1-10heteroalkylene.

In some embodiments, E³is optionally substituted C₁-C₆alkylene, O, S, or NR^N;

In some embodiments, E³is optionally substituted C₁-C₆alkylene. In some embodiments, E³is optionally substituted C₁-C₃alkylene. In some embodiments, E³is O, S, or NR^N.

In some embodiments, E³is C₁-C₆alkylene. In some embodiments, E³is C₁-C₃alkylene. In some embodiments, E³is O.

In some embodiments, E³is

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where a is 0, 1, 2, 3, 4, or 5.

In some embodiments, E³is

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In some embodiments, each R_Nis, independently, H or optionally substituted C_1-4alkyl.

In some embodiments, each R_Nis, independently, H or methyl.

In some embodiments, E is

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where a is 0, 1, 2, 3, 4, or 5.

In some embodiments, E¹is

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where a is 0, 1, 2, 3, 4, or 5.

In some embodiments, E¹is

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In some embodiments, E¹is

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In some embodiments, E¹is

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In some embodiments, E¹is

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where

b is 0, 1, 2, 3, 4, 5, or 6;

R^ais H, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, or optionally substituted C₃-C₆carbocyclyl;

R^bis H, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, or optionally substituted C₃-C₆carbocyclyl; and

R^cis H, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, or optionally substituted C₃-C₆carbocyclyl.

In some embodiments, E¹is

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In some embodiments, E¹is

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In some embodiments, E¹is

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In some embodiments, R^ais H or optionally substituted C₁-C₆alkyl. In some embodiments, R^bis H or optionally substituted C₁-C₆alkyl. In some embodiments, R^cis H or optionally substituted C₁-C₆alkyl.

In some embodiments, R^ais H or methyl. In some embodiments, R^bis H or methyl. In some embodiments, R^cis H or methyl.

In some embodiments, b is 0, 1, 2, or 3. In some embodiments, b is 0. In some embodiments, b is 1. In some embodiments, b is 2. In some embodiments, b is 3.

In some embodiments, E¹is

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In some embodiments, E¹is

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In some embodiments, E¹is

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In some embodiments, E¹is

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In some embodiments, E¹is

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In some embodiments, E¹is

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In some embodiments, E²is O, NR^w,

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wherein

c is 0, 1, 2, 3, 4, 5, 6, 7, or 8;

d is 0, 1, 2, or 3;

e is 0, 1, 2, 3, 4, 5, or 6;

f is 0, 1, 2, 3, or 4;

R^dis H, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, or optionally substituted C₃-C₆carbocyclyl;

R^eis H, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, or optionally substituted C₃-C₆carbocyclyl;

R^fis H, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, or optionally substituted C₃-C₆carbocyclyl;

R^gis H, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, or optionally substituted C₃-C₆carbocyclyl; and

W is O or NR^w, wherein R^wis H or optionally substituted C₁-C₆alkyl.

In some embodiments, E²is O, NR^w,

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In some embodiments, R^dis H or optionally substituted C₁-C₆alkyl. In some embodiments, Re is H or optionally substituted C₁-C₆alkyl. In some embodiments, R^fis H or optionally substituted C₁-C₆alkyl. In some embodiments, R^gis H or optionally substituted C₁-C₆alkyl. In some embodiments, R^wis H or optionally substituted C₁-C₆alkyl.

In some embodiments, R^dis H or methyl. In some embodiments, R^eis H or methyl. In some embodiments, R^fis H or methyl. In some embodiments, R^gis H or methyl. In some embodiments, R^wis H or methyl.

In some embodiments, E²is

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In some embodiments, E²is O,

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In some embodiments, each of F¹, F², or F³is, independently, optionally substituted C₃-C₁₀carbocyclylene.

In some embodiments, the C₃-C₁₀carbocyclylene is monocyclic. In some embodiments, the C₃-C₁₀carbocyclylene is polycyclic.

In some embodiments, the C₃-C₁₀carbocyclylene is bicyclic.

In some embodiments, the C₃-C₁₀carbocyclylene is bridged. In some embodiments, the C₃-C₁₀carbocyclylene is fused. In some embodiments, the C₃-C₁₀carbocyclylene is spirocyclic.

In some embodiments, the C₃-C₁₀carbocyclylene is

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In some embodiments, F²is

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In some embodiments, the C₃-C₁₀carbocyclylene is

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In some embodiments, F¹is

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In some embodiments, each of F¹, F², or F³is, independently, optionally substituted C₂-C₉heterocyclylene.

In some embodiments, the C₂-C₉heterocyclylene is monocyclic. In some embodiments, the C₂-C₉heterocyclylene is polycyclic.

In some embodiments, the C₂-C₉heterocyclylene is bicyclic.

In some embodiments, the C₂-C₉heterocyclylene is bridged. In some embodiments, the C₂-C₉heterocyclylene is fused. In some embodiments, the C₂-C₉heterocyclylene is spirocyclic.

In some embodiments, the C₂-C₉heterocyclylene includes a quaternary amine.

In some embodiments, the C₂-C₉heterocyclylene is

embedded image

where

q1 is 0, 1, 2, 3, or 4;

q2 is 0, 1, 2, 3, 4, 5, or 6;

q3 is 0, 1, 2, 3, 4, 5, 6, 7, or 8;

each R^his, independently, ²H, halogen, optionally substituted C₁-C₆alkyl, ORⁱ², or NRⁱ³Rⁱ⁴; or two R^hgroups, together with the carbon atom to which each is attached, combine to form optionally substituted C₃-C₁₀carbocyclyl or optionally substituted C₂-C₉heterocyclyl; or two R^hgroups, together with the carbon atoms to which each is attached, combine to form optionally substituted C₃-C₁₀carbocyclyl or optionally substituted C₂-C₉heterocyclyl;

Rⁱ¹is H or optionally substituted C₁-C₆alkyl;

Rⁱ²is H, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆heteroalkyl, or optionally substituted C₃-C₆carbocyclyl;

Rⁱ³is H or optionally substituted C₁-C₆alkyl; and

Rⁱ⁴is H or optionally substituted C₁-C₆alkyl.

In some embodiments, each R^his, independently, halogen, optionally substituted C₁-C₆alkyl, ORⁱ², or NRⁱ³Rⁱ⁴. In some embodiments, Rⁱ¹is H or optionally substituted C₁-C₆alkyl. In some embodiments, Rⁱ²is H or optionally substituted C₁-C₆alkyl. In some embodiments, Rⁱ³is H or optionally substituted C₁-C₆alkyl. In some embodiments, Rⁱ⁴is H or optionally substituted C₁-C₆alkyl.

In some embodiments, the C₂-C₉heterocyclylene is

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In some embodiments, each R^his, independently, halogen, optionally substituted C₁-C₆alkyl, ORⁱ², or NRⁱ³Rⁱ⁴. In some embodiments, each R^his, independently, halogen, optionally substituted C₁-C₆alkyl, or NRⁱ³Rⁱ⁴.

In some embodiments, each R^his, independently, ²H, halogen, cyano, optionally substituted C₁-C₆alkyl, ORⁱ², or NRⁱ³Rⁱ⁴. In some embodiments, two R^hgroups, together with the carbon atom to which each is attached, combine to form optionally substituted C₃-C₁₀carbocyclyl or optionally substituted C₂-C₉heterocyclyl. In some embodiments, two R^hgroups, together with the carbon atoms to which each is attached, combine to form optionally substituted C₃-C₁₀carbocyclyl or optionally substituted C₂-C₉heterocyclyl.

In some embodiments, each R^his, independently, ²H, F, methyl,

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In some embodiments, each R^his, independently, F, methyl, or NRⁱ³Rⁱ⁴.

In some embodiments, q1 is 0, 1, or 2. In some embodiments, q1 is 0. In some embodiments, q1 is 1. In some embodiments, q1 is 2.

In some embodiments, q2 is 0, 1, or 2. In some embodiments, q2 is 0. In some embodiments, q2 is 1. In some embodiments, q2 is 2.

In some embodiments, q3 is 0, 1, or 2. In some embodiments, q3 is 0. In some embodiments, q3 is 1. In some embodiments, q3 is 2.

In some embodiments, the C₂-C₉heterocyclylene is

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In some embodiments, the C₂-C₉heterocyclylene is

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In some embodiments, the C₂-C₉heterocyclylene is

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In some embodiments, the C₂-C₉heterocyclylene is

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In some embodiments, F¹is

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In some embodiments, F¹is

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In some embodiments, F¹is

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In some embodiments, F²is

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In some embodiments, F²is

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In some embodiments, F³is

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In some embodiments, F³is

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In some embodiments, Rⁱ¹is H or methyl. In some embodiments, Rⁱ²is H or methyl. In some embodiments, Rⁱ³is H or methyl. In some embodiments, Rⁱ⁴is H or methyl.

In some embodiments, the C₂-C₉heterocyclylene is

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In some embodiments, the C₂-C₉heterocyclylene is

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In some embodiments, the C₂-C₉heterocyclylene is

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In some embodiments, the C₂-C₉heterocyclylene is

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In some embodiments, the C₂-C₉heterocyclylene is

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In some embodiments, F¹is

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In some embodiments, F¹is

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In some embodiments, F¹is

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In some embodiments, F²is

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In some embodiments, the C₂-C₉heterocyclyl is

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In some embodiments, the C₂-C₉heterocyclyl is

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In some embodiments, the C₂-C₉heterocyclyl is

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In some embodiments, the C₂-C₉heterocyclyl is

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In some embodiments, F¹is

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In some embodiments, F¹is

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In some embodiments, F¹is

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In some embodiments, F¹is

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In some embodiments, F¹is

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In some embodiments, F²is

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In some embodiments, F²is

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In some embodiments, F²is

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In some embodiments, F²is

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In some embodiments, F³is

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In some embodiments, each of F¹, F², or F³is, independently, optionally substituted C₆-C₁₀arylene.

In some embodiments, the C₆-C₁₀arylene is

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In some embodiments, each of F¹, F², or F³is, independently, optionally substituted C₂-C₉heteroarylene.

In some embodiments, the C₂-C₉heteroarylene is

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In some embodiments, F²is

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In some embodiments, F²is

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In some embodiments, C³is

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In some embodiments, C³is

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In some embodiments, m is 1. In some embodiments, p is 1.

In some embodiments, the linker has the structure of

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In some embodiments, the linker has the structure of

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In some embodiments, the linker has the structure of:

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In some embodiments, the linker is absent.

In some embodiments, the linker is optionally substituted C₃-C₁₀carbocyclylene, optionally substituted C_2-10heterocyclylene, optionally substituted C₆-C₁₀arylene, or optionally substituted C₂-C₉heteroarylene.

In some embodiments, the linker is optionally substituted C₃-C₁₀carbocyclylene or optionally substituted C_2-10heterocyclylene. In some embodiments, the linker is optionally substituted C₆-C₁₀arylene or optionally substituted C₂-C₉heteroarylene.

In some embodiments, the linker is optionally substituted C_2-10heterocyclylene.

In some embodiments, the C₂-C₉heterocyclylene is monocyclic. In some embodiments, the C₂-C₉heterocyclylene is polycyclic.

In some embodiments, the C₂-C₉heterocyclylene is bicyclic.

In some embodiments, the C₂-C₉heterocyclylene is bridged. In some embodiments, the C₂-C₉heterocyclylene is fused. In some embodiments, the C₂-C₉heterocyclylene is spirocyclic.

In some embodiments, the linker has the structure of

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In some embodiments, the linker has the structure of

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In some embodiments, the compound has the structure of any one of compounds D1-D15 in Table 2A, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound has the structure of any one of compounds D16-D90 in Table 2B, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound has the structure of any one of compounds D91-D220 in Table 2C, or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound has the structure of any one of compounds D3, D10, D12, D13, and D15 in Table 2A, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound has the structure of any one of compounds D16-D29, D33, D35-D43, D46, D49, D50, D57, D60, D65, D66, D70, D72-D75, D77-D79, D83-D87, D89, and D90 in Table 2B, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound has the structure of any one of compounds D96-D101-D123-D136, D138-D141, D144, D146, D147, D149-D170, and D173-D219 in Table 2C, or a pharmaceutically acceptable salt thereof.

In an aspect, the disclosure features a compound having the structure of any one of compounds D1-D15 in Table 2A, or a pharmaceutically acceptable salt thereof.

In an aspect, the disclosure features a compound having the structure of any one of compounds D16-D90 in Table 2B, or a pharmaceutically acceptable salt thereof.

In an aspect, the disclosure features a compound having the structure of any one of compounds D91-D220 in Table 2C, or a pharmaceutically acceptable salt thereof.

TABLE 2A

Compounds D1-D15 of the Disclosure

Compound No.
Structure

D1

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D10

D11

D12

D13

D14

D15

TABLE 2B

Compounds D16-D90 of the Disclosure

Comp-

ound

No.
Structure

D16

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D17

D18

D19

D20

D21

D22

D23

D24

D25

D26

D27

D28

D29

D30

D31

D32

D33

D34

D35

D36

D37

D38

D39

D40

D41

D42

D43

D44

D45

D46

D47

D48

D49

D50

D51

D52

D53

D54

D55

D56

D57

D58

D59

D60

D61

D62

D63

D64

D65

D66

D67

D68

D69

D70

D71

D72

D73

D74

D75

D76

D77

D78

D79

D80

D81

D82

D83

D84

D85

D86

D87

D88

D89

D90

TABLE 2C

Compounds D91-D220 of the Disclosure

Compound No.
Structure

D91

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D92

D93

D94

D95

D96

D97

D98

D99

D100

D101

D102

D103

D104

D105

D106

D107

D108

D109

D110

D111

D112

D113

D114

D115

D116

D117

D118

D119

D120

D121

D122

D123

D124

D125

D126

D127

D128

D129

D130

D131

D132

D133

D134

D135

D136

D137

D138

D139

D140

D141

D142

D143

D144

D145

D146

D147

D148

D149

D150

D151

D152

D153

D154

D155

D156

D157

D158

D159

D160

D161

D162

D163

D164

D165

D166

D167

D168

D169

D170

D171

D172

D173

D174

D175

D176

D177

D178

D179

D180

D181

D182

D183

D184

D185

D186

D187

D188

D189

D190

D191

D192

D193

D194

D195

D196

D197

D198

D199

D200

D201

D202

D203

D204

D205

D206

D207

D208

D209

D210

D211

D212

D213

D214

D215

D216

D217

D218

D219

D220

In an aspect, the disclosure features a compound having the structure of any one of DD1 and DD2, or a pharmaceutically acceptable salt thereof.

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In another aspect, the disclosure features a pharmaceutical composition including any of the foregoing compounds, or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable excipient.

In an aspect, the disclosure features a method of inhibiting the level and/or activity of BRD9 in a cell, the method involving contacting the cell with an effective amount of any of the foregoing compounds, or pharmaceutically acceptable salts thereof, or a pharmaceutical composition thereof.

In another aspect, the disclosure features a method of reducing the level and/or activity of BRD9 in a cell, the method involving contacting the cell with an effective amount of any of the foregoing compounds, or pharmaceutically acceptable salts thereof, or a pharmaceutical composition thereof.

In some embodiments, the cell is a cancer cell.

In some embodiments, the cancer is a malignant, rhabdoid tumor, a CD8+ T-cell lymphoma, endometrial carcinoma, ovarian carcinoma, bladder cancer, stomach cancer, pancreatic cancer, esophageal cancer, prostate cancer, renal cell carcinoma, melanoma, colorectal cancer, a sarcoma (e.g., a soft tissue sarcoma, synovial sarcoma, Ewing's sarcoma, osteosarcoma, rhabdomyosarcoma, adult fibrosarcoma, alveolar soft-part sarcoma, angiosarcoma, clear cell sarcoma, desmoplastic small round cell tumor, epithelioid sarcoma, fibromyxoid sarcoma, gastrointestinal stromal tumor, Kaposi sarcoma, liposarcoma, leiomyosarcoma, malignant mesenchymoma malignant peripheral nerve sheath tumors, myxofibrosarcoma, low-grade rhabdomyosarcoma), non-small cell lung cancer (e.g., squamous or adenocarcinoma), stomach cancer, or breast cancer. In some embodiments, the cancer is a malignant, rhabdoid tumor, a CD8+ T-cell lymphoma, endometrial carcinoma, ovarian carcinoma, bladder cancer, stomach cancer, pancreatic cancer, esophageal cancer, prostate cancer, renal cell carcinoma, melanoma, or colorectal cancer. In some embodiments, the cancer is a sarcoma (e.g., synovial sarcoma or Ewing's sarcoma), non-small cell lung cancer (e.g., squamous or adenocarcinoma), stomach cancer, or breast cancer. In some embodiments, the cancer is sarcoma (e.g., synovial sarcoma or Ewing's sarcoma). In some embodiments, the sarcoma is synovial sarcoma.

In an aspect, the disclosure features a method of treating a BAF complex-related disorder in a subject in need thereof, the method involving administering to the subject an effective amount of any of the foregoing compounds, or pharmaceutically acceptable salts thereof, or a pharmaceutical composition thereof. In some embodiments, the BAF complex-related disorder is cancer. In some embodiments, the BAF complex-related disorder is infection.

In another aspect, the disclosure features a method of treating an SS18-SSX fusion protein-related disorder in a subject in need thereof, the method involving administering to the subject an effective amount of any of the foregoing compounds, or pharmaceutically acceptable salts thereof, or a pharmaceutical composition thereof. In some embodiments, the SS18-SSX fusion protein-related disorder is cancer. In some embodiments, the SS18-SSX fusion protein-related disorder is infection. In some embodiments of any of the foregoing methods, the SS18-SSX fusion protein is a SS18-SSX1 fusion protein, a SS18-SSX2 fusion protein, or a SS18-SSX4 fusion protein.

In yet another aspect, the disclosure features a method of treating a BRD9-related disorder in a subject in need thereof, the method involving administering to the subject an effective amount of any of the foregoing compounds, or pharmaceutically acceptable salts thereof, or a pharmaceutical composition thereof. In some embodiments, the BRD9-related disorder is cancer. In some embodiments, the BRD9-related disorder is infection.

In some embodiments, the cancer is squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, hepatocellular carcinomas, and renal cell carcinomas, cancer of the bladder, bowel, breast, cervix, colon, esophagus, head, kidney, liver, lung, neck, ovary, pancreas, prostate, and stomach; leukemias; benign and malignant lymphomas, particularly Burkitt's lymphoma and Non-Hodgkin's lymphoma; benign and malignant melanomas; myeloproliferative diseases; sarcomas, including Ewing's sarcoma, hemangiosarcoma, Kaposi's sarcoma, liposarcoma, myosarcomas, peripheral neuroepithelioma, synovial sarcoma, gliomas, astrocytomas, oligodendrogliomas, ependymomas, gliobastomas, neuroblastomas, ganglioneuromas, gangliogliomas, medulloblastomas, pineal cell tumors, meningiomas, meningeal sarcomas, neurofibromas, and Schwannomas; bowel cancer, breast cancer, prostate cancer, cervical cancer, uterine cancer, lung cancer, ovarian cancer, testicular cancer, thyroid cancer, astrocytoma, esophageal cancer, pancreatic cancer, stomach cancer, liver cancer, colon cancer, melanoma; carcinosarcoma, Hodgkin's disease, Wilms' tumor and teratocarcinomas. Additional cancers which may be treated using the disclosed compounds according to the present invention include, for example, acute granulocytic leukemia, acute lymphocytic leukemia (ALL), acute myelogenous leukemia (AML), adenocarcinoma, adenosarcoma, adrenal cancer, adrenocortical carcinoma, anal cancer, anaplastic astrocytoma, angiosarcoma, appendix cancer, astrocytoma, Basal cell carcinoma, B-Cell lymphoma, bile duct cancer, bladder cancer, bone cancer, bone marrow cancer, bowel cancer, brain cancer, brain stem glioma, breast cancer, triple (estrogen, progesterone and HER-2) negative breast cancer, double negative breast cancer (two of estrogen, progesterone and HER-2 are negative), single negative (one of estrogen, progesterone and HER-2 is negative), estrogen-receptor positive, HER2-negative breast cancer, estrogen receptor-negative breast cancer, estrogen receptor positive breast cancer, metastatic breast cancer, luminal A breast cancer, luminal B breast cancer, Her2-negative breast cancer, HER2-positive or negative breast cancer, progesterone receptor-negative breast cancer, progesterone receptor-positive breast cancer, recurrent breast cancer, carcinoid tumors, cervical cancer, cholangiocarcinoma, chondrosarcoma, chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), colon cancer, colorectal cancer, craniopharyngioma, cutaneous lymphoma, cutaneous melanoma, diffuse astrocytoma, ductal carcinoma in situ (DCIS), endometrial cancer, ependymoma, epithelioid sarcoma, esophageal cancer, ewing sarcoma, extrahepatic bile duct cancer, eye cancer, fallopian tube cancer, fibrosarcoma, gallbladder cancer, gastric cancer, gastrointestinal cancer, gastrointestinal carcinoid cancer, gastrointestinal stromal tumors (GIST), germ cell tumor glioblastoma multiforme (GBM), glioma, hairy cell leukemia, head and neck cancer, hemangioendothelioma, Hodgkin lymphoma, hypopharyngeal cancer, infiltrating ductal carcinoma (IDC), infiltrating lobular carcinoma (ILC), inflammatory breast cancer (IBC), intestinal Cancer, intrahepatic bile duct cancer, invasive/infiltrating breast cancer, Islet cell cancer, jaw cancer, Kaposi sarcoma, kidney cancer, laryngeal cancer, leiomyosarcoma, leptomeningeal metastases, leukemia, lip cancer, liposarcoma, liver cancer, lobular carcinoma in situ, low-grade astrocytoma, lung cancer, lymph node cancer, lymphoma, male breast cancer, medullary carcinoma, medulloblastoma, melanoma, meningioma, Merkel cell carcinoma, mesenchymal chondrosarcoma, mesenchymous, mesothelioma metastatic breast cancer, metastatic melanoma metastatic squamous neck cancer, mixed gliomas, monodermal teratoma, mouth cancer mucinous carcinoma, mucosal melanoma, multiple myeloma, Mycosis Fungoides, myelodysplastic syndrome, nasal cavity cancer, nasopharyngeal cancer, neck cancer, neuroblastoma, neuroendocrine tumors (NETs), non-Hodgkin's lymphoma, non-small cell lung cancer (NSCLC), oat cell cancer, ocular cancer, ocular melanoma, oligodendroglioma, oral cancer, oral cavity cancer, oropharyngeal cancer, osteogenic sarcoma, osteosarcoma, ovarian cancer, ovarian epithelial cancer ovarian germ cell tumor, ovarian primary peritoneal carcinoma, ovarian sex cord stromal tumor, Paget's disease, pancreatic cancer, papillary carcinoma, paranasal sinus cancer, parathyroid cancer, pelvic cancer, penile cancer, peripheral nerve cancer, peritoneal cancer, pharyngeal cancer, pheochromocytoma, pilocytic astrocytoma, pineal region tumor, pineoblastoma, pituitary gland cancer, primary central nervous system (CNS) lymphoma, prostate cancer, rectal cancer, renal cell carcinoma, renal pelvis cancer, rhabdomyosarcoma, salivary gland cancer, soft tissue sarcoma, bone sarcoma, sarcoma, sinus cancer, skin cancer, small cell lung cancer (SCLC), small intestine cancer, spinal cancer, spinal column cancer, spinal cord cancer, squamous cell carcinoma, stomach cancer, synovial sarcoma, T-cell lymphoma, testicular cancer, throat cancer, thymoma/thymic carcinoma, thyroid cancer, tongue cancer, tonsil cancer, transitional cell cancer, tubal cancer, tubular carcinoma, undiagnosed cancer, ureteral cancer, urethral cancer, uterine adenocarcinoma, uterine cancer, uterine sarcoma, vaginal cancer, vulvar cancer, T-cell lineage acute lymphoblastic leukemia (T-ALL), T-cell lineage lymphoblastic lymphoma (T-LL), peripheral T-cell lymphoma, Adult T-cell leukemia, Pre-B ALL, Pre-B lymphomas, large B-cell lymphoma, Burkitts lymphoma, B-cell ALL, Philadelphia chromosome positive ALL, Philadelphia chromosome positive CML, juvenile myelomonocytic leukemia (JMML), acute promyelocytic leukemia (a subtype of AML), large granular lymphocytic leukemia, Adult T-cell chronic leukemia, diffuse large B cell lymphoma, follicular lymphoma; Mucosa-Associated Lymphatic Tissue lymphoma (MALT), small cell lymphocytic lymphoma, mediastinal large B cell lymphoma, nodal marginal zone B cell lymphoma (NMZL); splenic marginal zone lymphoma (SMZL); intravascular large B-cell lymphoma; primary effusion lymphoma; or lymphomatoid granulomatosis; B-cell prolymphocytic leukemia; splenic lymphoma/leukemia, unclassifiable, splenic diffuse red pulp small B-cell lymphoma; lymphoplasmacytic lymphoma; heavy chain diseases, for example, Alpha heavy chain disease, Gamma heavy chain disease, Mu heavy chain disease, plasma cell myeloma, solitary plasmacytoma of bone; extraosseous plasmacytoma; primary cutaneous follicle center lymphoma, T cell/histocyte rich large B-cell lymphoma, DLBCL associated with chronic inflammation; Epstein-Barr virus (EBV)+ DLBCL of the elderly; primary mediastinal (thymic) large B-cell lymphoma, primary cutaneous DLBCL, leg type, ALK+ large B-cell lymphoma, plasmablastic lymphoma; large B-cell lymphoma arising in HHV8-associated multicentric, Castleman disease; B-cell lymphoma, unclassifiable, with features intermediate between diffuse large B-cell lymphoma, or B-cell lymphoma, unclassifiable, with features intermediate between diffuse large B-cell lymphoma and classical Hodgkin lymphoma.

In some embodiments, the infection is viral infection (e.g., an infection with a virus of the Retroviridae family such as the lentiviruses (e.g. Human immunodeficiency virus (HIV) and deltaretroviruses (e.g., human T cell leukemia virus I (HTLV-I), human T cell leukemia virus II (HTLV-II)); Hepadnaviridae family (e.g. hepatitis B virus (HBV)); Flaviviridae family (e.g. hepatitis C virus (HCV)); Adenoviridae family (e.g. Human Adenovirus); Herpesviridae family (e.g. Human cytomegalovirus (HCMV), Epstein-Barr virus, herpes simplex virus 1 (HSV-1), herpes simplex virus 2 (HSV-2), human herpesvirus 6 (HHV-6), Herpesvirus K*, CMV, varicella-zoster virus); Papillomaviridae family (e.g. Human Papillomavirus (HPV, HPV E1)); Parvoviridae family (e.g. Parvovirus B19); Polyomaviridae family (e.g. JC virus and BK virus); Paramyxoviridae family (e.g. Measles virus); or Togaviridae family (e.g. Rubella virus)). In some embodiments, the disorder is Coffin Siris, Neurofibromatosis (e.g., NF-1, NF-2, or Schwannomatosis), or Multiple Meningioma. In an aspect, the disclosure features a method of treating a cancer in a subject in need thereof, the method including administering to the subject an effective amount of any of the foregoing compounds, or pharmaceutically acceptable salts thereof, or any of the foregoing pharmaceutical compositions.

In another aspect, the disclosure features a method for treating a viral infection in a subject in need thereof. This method includes administering to the subject an effective amount of any of the foregoing compounds, or pharmaceutically acceptable salts thereof, or any of the foregoing pharmaceutical compositions. In some embodiments, the viral infection is an infection with a virus of the Retroviridae family such as the lentiviruses (e.g. Human immunodeficiency virus (HIV) and deltaretroviruses (e.g., human T cell leukemia virus I (HTLV-I), human T cell leukemia virus II (HTLV-II)); Hepadnaviridae family (e.g. hepatitis B virus (HBV)), Flaviviridae family (e.g. hepatitis C virus (HCV)), Adenoviridae family (e.g. Human Adenovirus), Herpesviridae family (e.g. Human cytomegalovirus (HCMV), Epstein-Barr virus, herpes simplex virus 1 (HSV-1), herpes simplex virus 2 (HSV-2), human herpesvirus 6 (HHV-6), Herpesvirus K*, CMV, varicella-zoster virus), Papillomaviridae family (e.g. Human Papillomavirus (HPV, HPV E1)), Parvoviridae family (e.g. Parvovirus B19), Polyomaviridae family (e.g. JC virus and BK virus), Paramyxoviridae family (e.g. Measles virus), Togaviridae family (e.g. Rubella virus).

In another embodiment of any of the foregoing methods, the method further includes administering to the subject an additional anticancer therapy (e.g., chemotherapeutic or cytotoxic agent or radiotherapy).

In particular embodiments, the additional anticancer therapy is: a chemotherapeutic or cytotoxic agent (e.g., doxorubicin or ifosfamide), a differentiation-inducing agent (e.g., retinoic acid, vitamin D, cytokines), a hormonal agent, an immunological agent, or an anti-angiogenic agent. Chemotherapeutic and cytotoxic agents include, but are not limited to, alkylating agents, cytotoxic antibiotics, antimetabolites, vinca alkaloids, etoposides, and others (e.g., paclitaxel, taxol, docetaxel, taxotere, cis-platinum). A list of additional compounds having anticancer activity can be found in L. Brunton, B. Chabner and B. Knollman (eds). Goodman and Gilman's The Pharmacological Basis of Therapeutics, Twelfth Edition, 2011, McGraw Hill Companies, New York, N.Y.

In particular embodiments, the compound of the invention and the additional anticancer therapy and any of the foregoing compounds or pharmaceutical compositions are administered within 28 days of each other (e.g., within 21, 14, 10, 7, 5, 4, 3, 2, or 1 days) or within 24 hours (e.g., 12, 6, 3, 2, or 1 hours; or concomitantly) each in an amount that together are effective to treat the subject.

Chemical Terms

The terminology employed herein is for the purpose of describing particular embodiments and is not intended to be limiting.

For any of the following chemical definitions, a number following an atomic symbol indicates that total number of atoms of that element that are present in a particular chemical moiety. As will be understood, other atoms, such as hydrogen atoms, or substituent groups, as described herein, may be present, as necessary, to satisfy the valences of the atoms. For example, an unsubstituted C₂alkyl group has the formula —CH₂CH₃. When used with the groups defined herein, a reference to the number of carbon atoms includes the divalent carbon in acetal and ketal groups but does not include the carbonyl carbon in acyl, ester, carbonate, or carbamate groups. A reference to the number of oxygen, nitrogen, or sulfur atoms in a heteroaryl group only includes those atoms that form a part of a heterocyclic ring.

Herein a phrase of the form “optionally substituted X” (e.g., optionally substituted alkyl) is intended to be equivalent to “X, wherein X is optionally substituted” (e.g., “alkyl, wherein said alkyl is optionally substituted”). It is not intended to mean that the feature “X” (e.g., alkyl) per se is optional. As described herein, certain compounds of interest may contain one or more “optionally substituted” moieties. In general, the term “substituted”, whether preceded by the term “optionally” or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent, e.g., any of the substituents or groups described herein. Unless otherwise indicated, an “optionally substituted” group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position. Combinations of substituents envisioned by the present disclosure are preferably those that result in the formation of stable or chemically feasible compounds. The term “stable”, as used herein, refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and, in certain embodiments, their recovery, purification, and use for one or more of the purposes disclosed herein.

The term “aliphatic,” as used herein, refers to a saturated or unsaturated, straight, branched, or cyclic hydrocarbon. “Aliphatic” is intended herein to include, but is not limited to, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, and cycloalkynyl moieties, and thus incorporates each of these definitions. In one embodiment, “aliphatic” is used to indicate those aliphatic groups having 1-20 carbon atoms. The aliphatic chain can be, for example, mono-unsaturated, di-unsaturated, tri-unsaturated, or polyunsaturated, or alkynyl. Unsaturated aliphatic groups can be in a cis or trans configuration. In one embodiment, the aliphatic group contains from 1 to about 12 carbon atoms, more generally from 1 to about 6 carbon atoms or from 1 to about 4 carbon atoms. In one embodiment, the aliphatic group contains from 1 to about 8 carbon atoms. In certain embodiments, the aliphatic group is C₁-C₂, C₁-C₃, C₁-C₄, C₁-C₅, or C₁-C₆. The specified ranges as used herein indicate an aliphatic group having each member of the range described as an independent species. For example, the term C₁-C₆aliphatic as used herein indicates a straight or branched alkyl, alkenyl, or alkynyl group having from 1, 2, 3, 4, 5, or 6 carbon atoms and is intended to mean that each of these is described as an independent species. For example, the term C₁-C₄aliphatic as used herein indicates a straight or branched alkyl, alkenyl, or alkynyl group having from 1, 2, 3, or 4 carbon atoms and is intended to mean that each of these is described as an independent species. In one embodiment, the aliphatic group is substituted with one or more functional groups that results in the formation of a stable moiety.

The term “heteroaliphatic,” as used herein, refers to an aliphatic moiety that contains at least one heteroatom in the chain, for example, an amine, carbonyl, carboxy, oxo, thio, phosphate, phosphonate, nitrogen, phosphorus, silicon, or boron atoms in place of a carbon atom. In one embodiment, the only heteroatom is nitrogen. In one embodiment, the only heteroatom is oxygen. In one embodiment, the only heteroatom is sulfur. “Heteroaliphatic” is intended herein to include, but is not limited to, heteroalkyl, heteroalkenyl, heteroalkynyl, heterocycloalkyl, heterocycloalkenyl, and heterocycloalkynyl moieties. In one embodiment, “heteroaliphatic” is used to indicate a heteroaliphatic group (cyclic, acyclic, substituted, unsubstituted, branched or unbranched) having 1-20 carbon atoms. In one embodiment, the heteroaliphatic group is optionally substituted in a manner that results in the formation of a stable moiety. Nonlimiting examples of heteroaliphatic moieties are polyethylene glycol, polyalkylene glycol, amide, polyamide, polylactide, polyglycolide, thioether, ether, alkyl-heterocycle-alkyl, —O-alkyl-O-alkyl, and alkyl-O-haloalkyl.

The term “acyl,” as used herein, represents a hydrogen or an alkyl group that is attached to a parent molecular group through a carbonyl group, as defined herein, and is exemplified by formyl (i.e., a carboxyaldehyde group), acetyl, trifluoroacetyl, propionyl, and butanoyl. Exemplary unsubstituted acyl groups include from 1 to 6, from 1 to 11, or from 1 to 21 carbons.

The term “alkyl,” as used herein, refers to a branched or straight-chain monovalent saturated aliphatic hydrocarbon radical 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). An “alkylene” is a divalent alkyl group.

The term “alkenyl,” as used herein, alone or in combination with other groups, refers to a straight chain or branched hydrocarbon residue having a carbon-carbon double bond and having 2 to 20 carbon atoms (e.g., 2 to 16 carbon atoms, 2 to 10 carbon atoms, 2 to 6, or 2 carbon atoms). An “alkenylene” is a divalent alkenyl group.

The term “alkynyl,” as used herein, alone or in combination with other groups, refers to a straight chain or branched hydrocarbon residue having a carbon-carbon triple bond and having 2 to 20 carbon atoms (e.g., 2 to 16 carbon atoms, 2 to 10 carbon atoms, 2 to 6, or 2 carbon atoms). An “alkynylene” is a divalent alkynyl group.

The term “amino,” as used herein, represents —N(R^N1)₂, wherein each R^N1is, independently, H, OH, NO₂, N(R^N2)₂, SO₂OR^N2, SO₂R^N2, SOR^N2, an N-protecting group, alkyl, alkoxy, aryl, arylalkyl, cycloalkyl, acyl (e.g., acetyl, trifluoroacetyl, or others described herein), wherein each of these recited R^N1groups can be optionally substituted; or two R^N1combine to form an alkylene or heteroalkylene, and wherein each R^N2is, independently, H, alkyl, or aryl. The amino groups of the compounds described herein can be an unsubstituted amino (i.e., —NH₂) or a substituted amino (i.e., —N(R^N1)₂).

The term “aryl,” as used herein, refers to an aromatic mono- or polycarbocyclic radical of, e.g., 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.

The term “arylalkyl,” as used herein, represents an alkyl group substituted with an aryl group. Exemplary unsubstituted arylalkyl groups are from 7 to 30 carbons (e.g., from 7 to 16 or from 7 to 20 carbons, such as C₁-C₆alkyl C₆-C₁₀aryl, C₁-C₁₀alkyl C₆-C₁₀aryl, or C₁-C₂₀alkyl C₆-C₁₀aryl), such as, benzyl and phenethyl. In some embodiments, the alkyl and the aryl each can be further substituted with 1, 2, 3, or 4 substituent groups as defined herein for the respective groups.

The term “azido,” as used herein, represents a —N₃group.

The term “bridged cyclyl,” as used herein, refers to a bridged polycyclic group of 5 to 20 atoms, containing from 1 to 3 bridges. Bridged cyclyl includes bridged carbocyclyl (e.g., norbornyl) and bridged heterocyclyl (e.g., 1,4-diazabicyclo[2.2.2]octane).

The term “cyano,” as used herein, represents a —CN group.

The term “carbocyclyl,” as used herein, refers to a non-aromatic C₃-C₁₂, monocyclic or polycyclic (e.g., bicyclic or tricyclic) structure in which the rings are formed by carbon atoms. Carbocyclyl structures include cycloalkyl groups (e.g., cyclohexyl) and unsaturated carbocyclyl radicals (e.g., cyclohexenyl). Polycyclic carbocyclyl includes spirocyclic carbocyclyl, bridged carbocyclyl, and fused carbocyclyl. A “carbocyclylene” is a divalent carbocyclyl group.

The term “cycloalkyl,” as used herein, refers to a saturated, non-aromatic, monovalent mono- or polycarbocyclic radical of 3 to 10, preferably 3 to 6 carbon atoms. This term is further exemplified by radicals such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, and adamantyl.

The terms “halo” or “halogen,” as used herein, mean a fluorine (fluoro), chlorine (chloro), bromine (bromo), or iodine (iodo) radical.

The term “heteroalkyl,” as used herein, refers to an alkyl group, as defined herein, in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen, or sulfur. In some embodiments, the heteroalkyl group can be further substituted with 1, 2, 3, or 4 substituent groups as described herein for alkyl groups. Examples of heteroalkyl groups are an “alkoxy” which, as used herein, refers to alkyl-O— (e.g., methoxy and ethoxy), and an “alkylamino” which, as used herein, refers to N(alkyl)R^Na, where R^Nais H or alkyl (e.g., methylamino). A “heteroalkylene” is a divalent heteroalkyl group.

The term “heteroalkenyl,” as used herein, refers to an alkenyl group, as defined herein, in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen, or sulfur. In some embodiments, the heteroalkenyl group can be further substituted with 1, 2, 3, or 4 substituent groups as described herein for alkenyl groups. Examples of heteroalkenyl groups are an “alkenoxy” which, as used herein, refers to alkenyl-O—. A “heteroalkenylene” is a divalent heteroalkenyl group.

The term “heteroalkynyl,” as used herein, refers to an alkynyl group, as defined herein, in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen, or sulfur. In some embodiments, the heteroalkynyl group can be further substituted with 1, 2, 3, or 4 substituent groups as described herein for alkynyl groups. Examples of heteroalkynyl groups are an “alkynoxy” which, as used herein, refers to alkynyl-O—. A “heteroalkynylene” is a divalent heteroalkynyl group.

The term “heteroaryl,” as used herein, refers to an aromatic monocyclic or polycyclic structure of 5 to 12 atoms having at least one aromatic ring containing 1, 2, or 3 ring atoms selected from nitrogen, oxygen, and sulfur, with the remaining ring atoms being carbon. One or two ring carbon atoms of the heteroaryl group may be replaced with a carbonyl group. Examples of heteroaryl groups are pyridyl, pyrazoyl, benzooxazolyl, benzoimidazolyl, benzothiazolyl, imidazolyl, oxazolyl, and thiazolyl. A “heteroarylene” is a divalent heteroaryl group.

The term “heteroarylalkyl,” as used herein, represents an alkyl group substituted with a heteroaryl group. Exemplary unsubstituted heteroarylalkyl groups are from 7 to 30 carbons (e.g., from 7 to 16 or from 7 to 20 carbons, such as C₁-C₆alkyl C₂-C₉heteroaryl, C₁-C₁₀alkyl C₂-C₉heteroaryl, or C₁-C₂₀alkyl C₂-C₉heteroaryl). In some embodiments, the alkyl and the heteroaryl each can be further substituted with 1, 2, 3, or 4 substituent groups as defined herein for the respective groups.

The term “heterocyclyl,” as used herein, refers a monocyclic or polycyclic radical (e.g., bicyclic or tricyclic) having 3 to 12 atoms having at least one non-aromatic ring containing 1, 2, 3, or 4 ring atoms selected from N, O, or S, and no aromatic ring containing any N, O, or S atoms. Polycyclic heterocyclyl includes spirocyclic heterocyclyl, bridged heterocyclyl, and fused heterocyclyl. Examples of heterocyclyl groups include, but are not limited to, morpholinyl, thiomorpholinyl, furyl, piperazinyl, piperidinyl, pyranyl, pyrrolidinyl, tetrahydropyranyl, tetrahydrofuranyl, and 1,3-dioxanyl. A “heterocyclylene” is a divalent heterocyclyl group.

The term “heterocyclylalkyl,” as used herein, represents an alkyl group substituted with a heterocyclyl group. Exemplary unsubstituted heterocyclylalkyl groups are from 7 to 30 carbons (e.g., from 7 to 16 or from 7 to 20 carbons, such as C₁-C₆alkyl C₂-C₉heterocyclyl, C₁-C₁₀alkyl C₂-C₉heterocyclyl, or C₁-C₂₀alkyl C₂-C₉heterocyclyl). In some embodiments, the alkyl and the heterocyclyl each can be further substituted with 1, 2, 3, or 4 substituent groups as defined herein for the respective groups.

The term “hydroxyalkyl,” as used herein, represents alkyl group substituted with an —OH group.

The term “hydroxyl,” as used herein, represents an —OH group.

The term “imine,” as used herein, represents ═NR^Ngroup, where R^Nis, e.g., H or alkyl.

The term “N-protecting group,” as used herein, represents those groups intended to protect an amino group against undesirable reactions during synthetic procedures. Commonly used N-protecting groups are disclosed in Greene, “Protective Groups in Organic Synthesis,” 3rd Edition (John Wiley & Sons, New York, 1999). N-protecting groups include, but are not limited to, acyl, aryloyl, or carbamyl groups such as formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthalyl, o-nitrophenoxyacetyl, α-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, 4-nitrobenzoyl, and chiral auxiliaries such as protected or unprotected D, L, or D, L-amino acids such as alanine, leucine, and phenylalanine; sulfonyl-containing groups such as benzenesulfonyl, and p-toluenesulfonyl; carbamate forming groups such as benzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl, 2,4-20 dimethoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl, 3,4,5-trimethoxybenzyloxycarbonyl, 1-(p-biphenylyl)-1-methylethoxycarbonyl, α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, benzhydryloxy carbonyl, t-butyloxycarbonyl, diisopropylmethoxycarbonyl, isopropyloxycarbonyl, ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl, 2,2,2,-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitrophenoxy carbonyl, 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, t-butylacetyl, alanyl, phenylsulfonyl, benzyl, t-butyloxycarbonyl (Boc), and benzyloxycarbonyl (Cbz).

The term “nitro,” as used herein, represents an —NO₂group.

The term “oxo,” as used herein, represents an ═O group.

The term “thiol,” as used herein, represents an —SH group.

The alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl (e.g., cycloalkyl), aryl, heteroaryl, and heterocyclyl groups may be substituted or unsubstituted. When substituted, there will generally be 1 to 4 substituents present, unless otherwise specified. Substituents include, for example: alkyl (e.g., unsubstituted and substituted, where the substituents include any group described herein, e.g., aryl, halo, hydroxy), aryl (e.g., substituted and unsubstituted phenyl), carbocyclyl (e.g., substituted and unsubstituted cycloalkyl), halogen (e.g., fluoro), hydroxyl, heteroalkyl (e.g., substituted and unsubstituted methoxy, ethoxy, or thioalkoxy), heteroaryl, heterocyclyl, amino (e.g., NH₂or mono- or dialkyl amino), azido, cyano, nitro, oxo, sulfonyl, or thiol. Aryl, carbocyclyl (e.g., cycloalkyl), heteroaryl, and heterocyclyl groups may also be substituted with alkyl (unsubstituted and substituted such as arylalkyl (e.g., substituted and unsubstituted benzyl)).

Compounds described herein (e.g., compounds of the invention) can have one or more asymmetric carbon atoms and can exist in the form of optically pure enantiomers, mixtures of enantiomers such as, for example, racemates, optically pure diastereoisomers, mixtures of diastereoisomers, diastereoisomeric racemates, or mixtures of diastereoisomeric racemates. The optically active forms can be obtained for example by resolution of the racemates, by asymmetric synthesis or asymmetric chromatography (chromatography with a chiral adsorbent or eluant). That is, certain of the disclosed compounds may exist in various stereoisomeric forms. Stereoisomers are compounds that differ only in their spatial arrangement. Enantiomers are pairs of stereoisomers whose mirror images are not superimposable, most commonly because they contain an asymmetrically substituted carbon atom that acts as a chiral center. “Enantiomer” means one of a pair of molecules that are mirror images of each other and are not superimposable. Diastereomers are stereoisomers that are not related as mirror images, most commonly because they contain two or more asymmetrically substituted carbon atoms and represent the configuration of substituents around one or more chiral carbon atoms. Enantiomers of a compound can be prepared, for example, by separating an enantiomer from a racemate using one or more well-known techniques and methods, such as, for example, chiral chromatography and separation methods based thereon. The appropriate technique and/or method for separating an enantiomer of a compound described herein from a racemic mixture can be readily determined by those of skill in the art. “Racemate” or “racemic mixture” means a compound containing two enantiomers, wherein such mixtures exhibit no optical activity; i.e., they do not rotate the plane of polarized light. “Geometric isomer” means isomers that differ in the orientation of substituent atoms in relationship to a carbon-carbon double bond, to a cycloalkyl ring, or to a bridged bicyclic system. Atoms (other than H) on each side of a carbon-carbon double bond may be in an 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 configurations relative to the core molecule. Certain of the disclosed compounds may exist in atropisomeric forms. Atropisomers are stereoisomers resulting from hindered rotation about single bonds where the steric strain barrier to rotation is high enough to allow for the isolation of the conformers. The compounds described herein (e.g., the compounds of the invention) may be prepared as individual isomers by either isomer-specific synthesis or resolved from an isomeric mixture. Conventional resolution techniques include forming the salt of a free base of each isomer of an isomeric pair using an optically active acid (followed by fractional crystallization and regeneration of the free base), forming the salt of the acid form of each isomer of an isomeric pair using an optically active amine (followed by fractional crystallization and regeneration of the free acid), forming an ester or amide of each of the isomers of an isomeric pair using an optically pure acid, amine or alcohol (followed by chromatographic separation and removal of the chiral auxiliary), or resolving an isomeric mixture of either a starting material or a final product using various well known chromatographic methods. When the stereochemistry of a disclosed compound is named or depicted by structure, the named or depicted stereoisomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by weight relative to the other stereoisomers. When a single enantiomer is named or depicted by structure, the depicted or named enantiomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by weight optically pure. When a single diastereomer is named or depicted by structure, the depicted or named diastereomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by weight pure. Percent optical purity is the ratio of the weight of the enantiomer or over the weight of the enantiomer plus the weight of its optical isomer. Diastereomeric purity by weight is the ratio of the weight of one diastereomer or over the weight of all the diastereomers. When the stereochemistry of a disclosed compound is named or depicted by structure, the named or depicted stereoisomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by mole fraction pure relative to the other stereoisomers. When a single enantiomer is named or depicted by structure, the depicted or named enantiomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by mole fraction pure. When a single diastereomer is named or depicted by structure, the depicted or named diastereomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by mole fraction pure. Percent purity by mole fraction is the ratio of the moles of the enantiomer or over the moles of the enantiomer plus the moles of its optical isomer. Similarly, percent purity by moles fraction is the ratio of the moles of the diastereomer or over the moles of the diastereomer plus the moles of its isomer. When a disclosed compound is named or depicted by structure without indicating the stereochemistry, and the compound has at least one chiral center, it is to be understood that the name or structure encompasses either enantiomer of the compound free from the corresponding optical isomer, a racemic mixture of the compound, or mixtures enriched in one enantiomer relative to its corresponding optical isomer. When a disclosed compound is named or depicted by structure without indicating the stereochemistry and has two or more chiral centers, it is to be understood that the name or structure encompasses a diastereomer free of other diastereomers, a number of diastereomers free from other diastereomeric pairs, mixtures of diastereomers, mixtures of diastereomeric pairs, mixtures of diastereomers in which one diastereomer is enriched relative to the other diastereomer(s), or mixtures of diastereomers in which one or more diastereomer is enriched relative to the other diastereomers. The invention embraces all of these forms.

Compounds of the present disclosure also include all of the isotopes of the atoms occurring in the intermediate or final compounds. “Isotopes” refers to atoms having the same atomic number but different mass numbers resulting from a different number of neutrons in the nuclei. For example, isotopes of hydrogen include tritium and deuterium.

Unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. Exemplary isotopes that can be incorporated into compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, such as ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹³N, ¹⁵N, ¹⁵O, ¹⁷O, ¹⁸O, ³²P, ³³P, ³⁸S, ¹⁸F, ³⁶Cl, ¹²³I and ¹²⁵I. Isotopically-labeled compounds (e.g., those labeled with ³H and ¹⁴C) can be useful in compound or substrate tissue distribution assays. Tritiated (i.e., ³H) and carbon-14 (i.e., ¹⁴C) isotopes can be useful for their ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium (i.e., ²H) may afford certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements). In some embodiments, one or more hydrogen atoms are replaced by ²H or ³H, or one or more carbon atoms are replaced by ¹³C- or ¹⁴C-enriched carbon. Positron emitting isotopes such as ¹⁵O, ¹³N, ¹¹C, and ¹⁸F are useful for positron emission tomography (PET) studies to examine substrate receptor occupancy. Preparations of isotopically labelled compounds are known to those of skill in the art. For example, isotopically labeled compounds can generally be prepared by following procedures analogous to those disclosed for compounds of the present invention described herein, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

As is known in the art, many chemical entities can adopt a variety of different solid forms such as, for example, amorphous forms or crystalline forms (e.g., polymorphs, hydrates, solvate). In some embodiments, compounds of the present invention may be utilized in any such form, including in any solid form. In some embodiments, compounds described or depicted herein may be provided or utilized in hydrate or solvate form.

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

Definitions

In this application, unless otherwise clear from 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 “including” and “including” may be understood to encompass itemized components or steps whether presented by themselves or together with one or more additional components or steps.

As used herein, the terms “about” and “approximately” refer to a value that is within 10% above or below the value being described. For example, the term “about 5 nM” indicates a range of from 4.5 to 5.5 nM.

As used herein, the term “administration” refers to the administration of a composition (e.g., a compound or a preparation that includes a compound as described herein) to a subject or system. Administration to an animal subject (e.g., to a human) may be by any appropriate route. For example, in some embodiments, administration may be bronchial (including by bronchial instillation), buccal, enteral, interdermal, intra-arterial, 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 vitreal.

As used herein, the term “adult soft tissue sarcoma” refers to a sarcoma that develops in the soft tissues of the body, typically in adolescent and adult subjects (e.g., subjects who are at least 10 years old, 11 years old, 12 years old, 13 years old, 14 years old, 15 years old, 16 years old, 17 years old, 18 years old, or 19 years old). Non-limiting examples of adult soft tissue sarcoma include, but are not limited to, synovial sarcoma, fibrosarcoma, malignant fibrous histiocytoma, dermatofibrosarcoma, liposarcoma, leiomyosarcoma, hemangiosarcoma, Kaposi's sarcoma, lymphangiosarcoma, malignant peripheral nerve sheath tumor/neurofibrosarcoma, extraskeletal chondrosarcoma, extraskeletal osteosarcoma, extraskeletal myxoid chondrosarcoma, and extraskeletal mesenchymal.

The term “antisense,” as used herein, refers to a nucleic acid comprising a polynucleotide that is sufficiently complementary to all or a portion of a gene, primary transcript, or processed mRNA, so as to interfere with expression of the endogenous gene (e.g., BRD9). “Complementary” polynucleotides are those that are capable of base pairing according to the standard Watson-Crick complementarity rules. Specifically, purines will base pair with pyrimidines to form a combination of guanine paired with cytosine (G:C) and adenine paired with either thymine (A:T) in the case of DNA, or adenine paired with uracil (A:U) in the case of RNA. It is understood that two polynucleotides may hybridize to each other even if they are not completely complementary to each other, provided that each has at least one region that is substantially complementary to the other.

The term “antisense nucleic acid” includes single-stranded RNA as well as double-stranded DNA expression cassettes that can be transcribed to produce an antisense RNA. “Active” antisense nucleic acids are antisense RNA molecules that are capable of selectively hybridizing with a primary transcript or mRNA encoding a polypeptide having at least 80% sequence identity (e.g., 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.9% identity, or more) with the targeted polypeptide sequence (e.g., a BRD9 polypeptide sequence). The antisense nucleic acid can be complementary to an entire coding strand, or to only a portion thereof. In some embodiments, an antisense nucleic acid molecule is antisense to a “coding region” of the coding strand of a nucleotide sequence. The term “coding region” refers to the region of the nucleotide sequence comprising codons that are translated into amino acid residues. In some embodiments, the antisense nucleic acid molecule is antisense to a “noncoding region” of the coding strand of a nucleotide sequence. The term “noncoding region” refers to 5′ and 3′ sequences that flank the coding region that are not translated into amino acids (i.e., also referred to as 5′ and 3′ untranslated regions). The antisense nucleic acid molecule can be complementary to the entire coding region of mRNA, or can be antisense to only a portion of the coding or noncoding region of an mRNA. For example, the antisense oligonucleotide can be complementary to the region surrounding the translation start site. An antisense oligonucleotide can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides in length.

As used herein, the term “BAF complex” refers to the BRG1- or HRBM-associated factors complex in a human cell.

As used herein, the term “BAF complex-related disorder” refers to a disorder that is caused or affected by the level and/or activity of a BAF complex.

As used herein, the terms “GBAF complex” and “GBAF” refer to a SWI/SNF ATPase chromatin remodeling complex in a human cell. GBAF complex subunits may include, but are not limited to, ACTB, ACTL6A, ACTL6B, BICRA, BICRAL, BRD9, SMARCA2, SMARCA4, SMARCC1, SMARCD1, SMARCD2, SMARCD3, and SS18. The term “cancer” refers to a condition caused by the proliferation of malignant neoplastic cells, such as tumors, neoplasms, carcinomas, sarcomas, leukemias, and lymphomas.

As used herein, the term “BRD9” refers to bromodomain-containing protein 9, a component of the BAF (BRG1- or BRM-associated factors) complex, a SWI/SNF ATPase chromatin remodeling complex, and belongs to family IV of the bromodomain-containing proteins. BRD9 is encoded by the BRD9 gene, the nucleic acid sequence of which is set forth in SEQ ID NO: 1. The term “BRD9” also refers to natural variants of the wild-type BRD9 protein, such as proteins having at least 85% identity (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.9% identity, or more) to the amino acid sequence of wild-type BRD9, which is set forth in SEQ ID NO: 2.

As used herein, the term “BRD9-related disorder” refers to a disorder that is caused or affected by the level and/or activity of BRD9. The term “cancer” refers to a condition caused by the proliferation of malignant neoplastic cells, such as tumors, neoplasms, carcinomas, sarcomas, leukemias, and lymphomas.

As used herein, a “combination therapy” or “administered in combination” means that two (or more) different agents or treatments are administered to a subject as part of a defined treatment regimen for a particular disease or condition. The treatment regimen defines the doses and periodicity of administration of each agent such that the effects of the separate agents on the subject overlap. In some embodiments, the delivery of the two or more agents is simultaneous or concurrent and the agents may be co-formulated. In some embodiments, the two or more agents are not co-formulated and are administered in a sequential manner as part of a prescribed regimen. In some embodiments, administration of two or more agents or treatments in combination is such that the reduction in a symptom, or other parameter related to the disorder is greater than what would be observed with one agent or treatment delivered alone or in the absence of the other. The effect of the two treatments can be partially additive, wholly additive, or greater than additive (e.g., synergistic). Sequential or substantially simultaneous administration of each therapeutic agent can be effected by any appropriate route including, but not limited to, oral routes, intravenous routes, intramuscular routes, and direct absorption through mucous membrane tissues. The therapeutic agents can be administered by the same route or by different routes. For example, a first therapeutic agent of the combination may be administered by intravenous injection while a second therapeutic agent of the combination may be administered orally.

A “compound of the present invention” and similar terms as used herein, whether explicitly noted or not, refers to compounds useful for treating BAF-related disorders (e.g., cancer or infection) described herein, including, e.g., compounds of Formula I or Formula II (e.g., compounds of Table 2A, Table 2B, and Table 2C), as well as salts (e.g., pharmaceutically acceptable salts), solvates, hydrates, stereoisomers (including atropisomers), and tautomers thereof. Those skilled in the art will appreciate that certain compounds described herein can exist in one or more different isomeric (e.g., stereoisomers, geometric isomers, atropisomers, and tautomers) or isotopic (e.g., in which one or more atoms has been substituted with a different isotope of the atom, such as hydrogen substituted for deuterium) forms. Unless otherwise indicated or clear from context, a depicted structure can be understood to represent any such isomeric or isotopic form, individually or in combination. Compounds described herein can be asymmetric (e.g., having one or more stereocenters). All stereoisomers, such as enantiomers and diastereomers, are intended unless otherwise indicated. Compounds of the present disclosure that contain asymmetrically substituted carbon atoms can be isolated in optically active or racemic forms. Methods on how to prepare optically active forms from optically active starting materials are known in the art, such as by resolution of racemic mixtures or by stereoselective synthesis. Many geometric isomers of olefins, C═N double bonds, and the like can also be present in the compounds described herein, and all such stable isomers are contemplated in the present disclosure. Cis and trans geometric isomers of the compounds of the present disclosure are described and may be isolated as a mixture of isomers or as separated isomeric forms. In some embodiments, one or more compounds depicted herein may exist in different tautomeric forms. As will be clear from context, unless explicitly excluded, references to such compounds encompass all such tautomeric forms. In some embodiments, tautomeric forms result from the swapping of a single bond with an adjacent double bond and the concomitant migration of a proton. In certain embodiments, a tautomeric form may be a prototropic tautomer, which is an isomeric protonation states having the same empirical formula and total charge as a reference form. Examples of moieties with prototropic tautomeric forms are ketone-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, amide-imidic acid pairs, enamine-imine pairs, and annular forms where a proton can occupy two or more positions of a heterocyclic system, such as, 1H- and 3H-imidazole, 1H-, 2H- and 4H-1,2,4-triazole, 1H- and 2H-isoindole, and 1H- and 2H-pyrazole. In some embodiments, tautomeric forms can be in equilibrium or sterically locked into one form by appropriate substitution. In certain embodiments, tautomeric forms result from acetal interconversion.

As used herein, the term “degrader” refers to a small molecule compound including a degradation moiety, wherein the compound interacts with a protein (e.g., BRD9) in a way which results in degradation of the protein, e.g., binding of the compound results in at least 5% reduction of the level of the protein, e.g., in a cell or subject.

As used herein, the term “degradation moiety” refers to a moiety whose binding results in degradation of a protein, e.g., BRD9. In one example, the moiety binds to a protease or a ubiquitin ligase that metabolizes the protein, e.g., BRD9.

By “determining the level of a protein” is meant the detection of a protein, or an mRNA encoding the protein, by methods known in the art either directly or indirectly. “Directly determining” means performing a process (e.g., performing an assay or test on a sample or “analyzing a sample” as that term is defined herein) to obtain the physical entity or value. “Indirectly determining” refers to receiving the physical entity or value from another party or source (e.g., a third-party laboratory that directly acquired the physical entity or value). Methods to measure protein level generally include, but are not limited to, western blotting, immunoblotting, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunoprecipitation, immunofluorescence, surface plasmon resonance, chemiluminescence, fluorescent 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 a property of a protein including, but not limited to, enzymatic activity or interaction with other protein partners. Methods to measure mRNA levels are known in the art.

As used herein, the terms “effective amount,” “therapeutically effective amount,” and “a “sufficient amount” of an agent that reduces the level and/or activity of BRD9 (e.g., in a cell or a subject) described herein refer to a quantity sufficient to, when administered to the subject, including a human, effect beneficial or desired results, including clinical results, and, as such, an “effective amount” or synonym thereto depends on the context in which it is being applied. For example, in the context of treating cancer, it is an amount of the agent that reduces the level and/or activity of BRD9 sufficient to achieve a treatment response as compared to the response obtained without administration of the agent that reduces the level and/or activity of BRD9. The amount of a given agent that reduces the level and/or activity of BRD9 described herein that will correspond to such an amount will vary depending upon various factors, such as the given agent, the pharmaceutical formulation, the route of administration, the type of disease or disorder, the identity of the subject (e.g., age, sex, and/or weight) or host being treated, and the like, but can nevertheless be routinely determined by one of skill in the art. Also, as used herein, a “therapeutically effective amount” of an agent that reduces the level and/or activity of BRD9 of the present disclosure is an amount which results in a beneficial or desired result in a subject as compared to a control. As defined herein, a therapeutically effective amount of an agent that reduces the level and/or activity of BRD9 of the present disclosure may be readily determined by one of ordinary skill by routine methods known in the art. Dosage regimen may be adjusted to provide the optimum therapeutic response.

As used herein, the term “inhibitor” refers to any agent which reduces the level and/or activity of a protein (e.g., BRD9). Non-limiting examples of inhibitors include small molecule inhibitors, degraders, antibodies, enzymes, or polynucleotides (e.g., siRNA).

The term “inhibitory RNA agent” refers to an RNA, or analog thereof, having sufficient sequence complementarity to a target RNA to direct RNA interference. Examples also include a DNA that can be used to make the RNA. RNA interference (RNAi) refers to a sequence-specific or selective process by which a target molecule (e.g., a target gene, protein, or RNA) is down-regulated. Generally, an interfering RNA (“iRNA”) is a double-stranded short-interfering RNA (siRNA), short hairpin RNA (shRNA), or single-stranded micro-RNA (miRNA) that results in catalytic degradation of specific mRNAs, and also can be used to lower or inhibit gene expression.

By “level” is meant a level of a protein, or mRNA encoding the protein, as compared to a reference. The reference can be any useful reference, as defined herein. By a “decreased level” or an “increased level” of a protein is meant a decrease or increase in protein level, as compared to a reference (e.g., a decrease or an increase by 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 an increase of more than about 10%, about 15%, about 20%, about 50%, about 75%, about 100%, or about 200%, as compared to a reference; a decrease or an increase by 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 by 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). A level of a protein may be expressed in mass/vol (e.g., g/dL, mg/mL, μg/mL, ng/mL) or percentage relative to total protein or mRNA in a sample.

The terms “miRNA” and “microRNA” refer to an RNA agent, preferably a single-stranded agent, of about 10-50 nucleotides in length, preferably between about 15-25 nucleotides in length, which is capable of directing or mediating RNA interference. Naturally-occurring miRNAs are generated from stem-loop precursor RNAs (i.e., pre-miRNAs) by Dicer. The term “Dicer” as used herein, includes Dicer as well as any Dicer ortholog or homolog capable of processing dsRNA structures into siRNAs, miRNAs, siRNA-like or miRNA-like molecules. The term microRNA (“miRNA”) is used interchangeably with the term “small temporal RNA” (“stRNA”) based on the fact that naturally-occurring miRNAs have been found to be expressed in a temporal fashion (e.g., during development).

By “modulating the activity of a BAF complex,” is meant altering the level of an activity related to a BAF complex (e.g., GBAF), or a related downstream effect. The activity level of a BAF complex may be measured using any method known in the art, e.g., the methods described in Kadoch et al, Cell 153:71-85 (2013), the methods of which are herein incorporated by reference.

“Percent (%) sequence identity” with respect to a reference polynucleotide or polypeptide sequence is defined as the percentage of nucleic acids or amino acids in a candidate sequence that are identical to the nucleic acids or amino acids in the reference polynucleotide or polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity. Alignment for purposes of determining percent nucleic acid or amino acid sequence identity can be achieved in various ways that are within the capabilities of one of skill in the art, for example, using publicly available computer software such as BLAST, BLAST-2, or Megalign software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared. For example, percent sequence identity values may be generated using the sequence comparison computer program BLAST. As an illustration, the percent sequence identity of a given nucleic acid or amino acid sequence, A, to, with, or against a given nucleic acid or amino acid sequence, B, (which can alternatively be phrased as a given nucleic acid or amino acid sequence, A that has a certain percent sequence identity to, with, or against a given nucleic acid or amino acid sequence, B) is calculated as follows:

100 multiplied by (the fraction X/Y)

where X is the number of nucleotides or amino acids scored as identical matches by a sequence alignment program (e.g., BLAST) in that program's alignment of A and B, and where Y is the total number of nucleic acids in B. It will be appreciated that where the length of nucleic acid or amino acid sequence A is not equal to the length of nucleic acid or amino acid sequence B, the percent sequence identity of A to B will not equal the percent sequence identity of B to A.

A “pharmaceutically acceptable excipient,” as used herein, refers any ingredient other than the compounds described herein (for example, a vehicle capable of suspending or dissolving the active compound) and having the properties of being substantially nontoxic and non-inflammatory in a patient. Excipients may include, for example: antiadherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, sorbents, suspending or dispersing agents, sweeteners, and waters of hydration. Exemplary excipients include, but are not limited to: butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, crosslinked polyvinyl pyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropyl cellulose, hydroxypropyl methylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methyl paraben, microcrystalline cellulose, polyethylene glycol, polyvinyl pyrrolidone, povidone, pregelatinized starch, propyl paraben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol.

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

The compounds described herein may have ionizable groups so as to be capable of preparation as pharmaceutically acceptable salts. These salts may be acid addition salts involving inorganic or organic acids or the salts may, in the case of acidic forms of the compounds described herein, be prepared from inorganic or organic bases. Frequently, the compounds are prepared or used as pharmaceutically acceptable salts prepared as addition products of pharmaceutically acceptable acids or bases. Suitable pharmaceutically acceptable acids and bases and methods for preparation of the 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. Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, and valerate salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium, as well as nontoxic ammonium, quaternary ammonium, and amine cations, including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, and ethylamine.

The term “pharmaceutical composition,” as used herein, represents a composition containing a compound described herein formulated with a pharmaceutically acceptable excipient, and manufactured or sold with the approval of a governmental regulatory agency as part of a therapeutic regimen for the treatment of disease in a mammal. Pharmaceutical compositions can be formulated, for example, for oral administration in unit dosage form (e.g., a tablet, capsule, caplet, gelcap, or syrup); for topical administration (e.g., as a cream, gel, lotion, or ointment); for intravenous administration (e.g., as a sterile solution free of particulate emboli and in a solvent system suitable for intravenous use); or in any other pharmaceutically acceptable formulation.

By “reducing the activity of BRD9,” is meant decreasing the level of an activity related to an BRD9, or a related downstream effect. A non-limiting example of inhibition of an activity of BRD9 is decreasing the level of a BAF complex (e.g., GBAF) in a cell. The activity level of BRD9 may be measured using any method known in the art. In some embodiments, an agent which reduces the activity of BRD9 is a small molecule BRD9 inhibitor. In some embodiments, an agent which reduces the activity of BRD9 is a small molecule BRD9 degrader.

By “reducing the level of BRD9,” is meant decreasing the level of BRD9 in a cell or subject. The level of BRD9 may be measured using any method known in the art.

By a “reference” is meant any useful reference used to compare protein or mRNA levels. The reference can be any sample, standard, standard curve, or level that is used for comparison purposes. The reference can be a normal reference sample or a reference standard or level. A “reference sample” can be, for example, a control, e.g., a predetermined negative control value such as a “normal control” or a prior sample taken from the same subject; a sample from a normal healthy subject, such as a normal cell or normal tissue; a sample (e.g., a cell or tissue) from a subject not having a disease; a sample from a subject that is diagnosed with a disease, but not yet treated with a compound described herein; a sample from a subject that has been treated by a compound described herein; or a sample of a purified protein (e.g., any described herein) at a known normal concentration. By “reference standard or level” is meant a value or number derived from a reference sample. A “normal control value” is a pre-determined value indicative of non-disease state, e.g., a value expected in a healthy control subject. Typically, a normal control value is expressed as a range (“between X and Y”), a high threshold (“no higher than X”), or a low threshold (“no lower than X”). A subject having a measured value within the normal control value for a particular biomarker is typically referred to as “within normal limits” for that biomarker. A normal reference standard or level can be a value or number derived from a normal subject not having a disease or disorder (e.g., cancer); a subject that has been treated with a compound described herein. In preferred embodiments, the reference sample, standard, or level is matched to the sample subject sample by at least one of the following criteria: age, weight, sex, disease stage, and overall health. A standard curve of levels of a purified protein, e.g., any described herein, within the normal reference range can also be used as a reference.

The terms “short interfering RNA” and “siRNA” (also known as “small interfering RNAs”) refer to an RNA agent, preferably a double-stranded agent, of about 10-50 nucleotides in length, the strands optionally having overhanging ends comprising, for example 1, 2 or 3 overhanging nucleotides (or nucleotide analogs), which is capable of directing or mediating RNA interference. Naturally-occurring siRNAs are generated from longer dsRNA molecules (e.g., >25 nucleotides in length) by a cell's RNAi machinery (e.g., Dicer or a homolog thereof).

The term “shRNA”, as used herein, refers to an RNA agent having a stem-loop structure, comprising a first and second region of complementary sequence, the degree of complementarity and orientation of the regions being sufficient such that base pairing occurs between the regions, the first and second regions being joined by a loop region, the loop resulting from a lack of base pairing between nucleotides (or nucleotide analogs) within the loop region.

As used herein, the term “subject” refers to any organism to which a composition in accordance with the invention may 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 seek or be in need of treatment, require treatment, be receiving treatment, be receiving treatment in the future, or be a human or animal who is under care by a trained professional for a particular disease or condition.

As used herein, the term “SS18-SSX fusion protein-related disorder” refers to a disorder that is caused or affected by the level and/or activity of SS18-SSX fusion protein.

As used herein, the terms “treat,” “treated,” or “treating” mean both therapeutic treatment and prophylactic or preventative measures wherein the object is to prevent or slow down (lessen) an undesired physiological condition, disorder, or disease, or obtain beneficial or desired clinical results. Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms; diminishment of the extent of a condition, disorder, or disease; stabilized (i.e., not worsening) state of condition, disorder, or disease; delay in onset or slowing of condition, disorder, or disease progression; amelioration of the condition, disorder, or disease state or remission (whether partial or total), whether detectable or undetectable; an amelioration of at least one measurable physical parameter, not necessarily discernible by the patient; or enhancement or improvement of condition, disorder, or disease. Treatment includes eliciting a clinically significant response without excessive levels of side effects. Treatment also includes prolonging survival as compared to expected survival if not receiving treatment.

As used herein, the terms “variant” and “derivative” are used interchangeably and refer to naturally-occurring, synthetic, and semi-synthetic analogues of a compound, peptide, protein, or other substance described herein. A variant or derivative of a compound, peptide, protein, or other substance described herein may retain or improve upon the biological activity of the original material.

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

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a series of graphs illustrating the effect of specific guide RNA (sgRNA) targeting of the BRD9 BAF complex subunit on synovial sarcoma cell growth. The Y-axis indicated the dropout ratio. The X-axis indicates the nucleotide position of the BRD9 gene. The grey box indicates the range of the negative control sgRNAs in the screen. The SYO1 cell line carries SS18-SSX2 fusion protein. The breakpoint joining the N-terminal region of SS18 to the C-terminal region of SSX2 are indicated by the black lines in their respective panel. The linear protein sequence is show with BRD9 PFAM domains annotated from the PFAM database.

FIG. 2 is an image illustrating dose dependent depletion of BRD9 levels in a synovial sarcoma cell line (SYO1) in the presence of a BRD9 degrader.

FIG. 3 is an image illustrating sustained suppression of BRD9 levels in a synovial sarcoma cell line (SYO1) in the presence of a BRD9 degrader over 72 hours.

FIG. 4 is an image illustrating sustained suppression of BRD9 levels in two cell lines (293T and SYO1) in the presence of a BRD9 degrader over 5 days.

FIG. 5 is an image illustrating sustained suppression of BRD9 levels in synovial sarcoma cell lines (SYO1 and Yamato) in the presence of a BRD9 degrader over 7 days compared to the levels in cells treated with CRISPR reagents.

FIG. 6 is an image illustrating the effect on cell growth of six cell lines (SYO1, Yamato, A549, HS-SY-II, ASKA, and 293T) in the presence of a BRD9 degrader and a BRD9 inhibitor.

FIG. 7 is an image illustrating the effect on cell growth of two cell lines (SYO1 and G401) in the presence of a BRD9 degrader.

FIG. 8 is an image illustrating the effect on cell growth of three synovial sarcoma cell lines (SYO1, HS-SY-II, and ASKA) in the presence of a BRD9 degrader, BRD9 binder and E3 ligase binder.

FIG. 9 is an image illustrating the effect on cell growth of three non-synovial sarcoma cell lines (RD, HCT116, and Calu6) in the presence of a BRD9 degrader, BRD9 binder and E3 ligase binder.

FIG. 10 is a graph illustrating the percentage of SYO1 in various cell cycle phases following treatment with DMSO, Compound 1 at 200 nM, or Compound 1 at 1 μM for 8 or 13 days.

FIG. 11 is a series of contour plots illustrating the percentage of SYO1 cells in various cell cycle phases following treatment with DMSO, Compound 1 at 200 nM, Compound 1 at 1 μM, or lenalidomide at 200 nM for 8 days. Numerical values corresponding to each contour plot are found in the table below.

FIG. 12 is a series of contour plots illustrating the percentage of SYO1 cells in various cell cycle phases following treatment with DMSO, Compound 1 at 200 nM, Compound 1 at 1 μM, or lenalidomide at 200 nM for 13 days. Numerical values corresponding to each contour plot are found in the table below.

FIG. 13 is a series of contour plots illustrating the percentage of early- and late-apoptotic SYO1 cells following treatment with DMSO, Compound 1 at 200 nM, Compound 1 at 1 μM, or lenalidomide at 200 nM for 8 days. Numerical values corresponding to each contour plot are found in the table below.

FIG. 14 is a graph illustrating the proteins present in BAF complexes including the SS18-SSX fusion protein.

DETAILED DESCRIPTION

The present disclosure features compositions and methods useful for the treatment of BAF-related disorders (e.g., cancer and infection). The disclosure further features compositions and methods useful for inhibition of the level and/or activity of BRD9, e.g., for the treatment of disorders such as cancer (e.g., sarcoma) and infection (e.g., viral infection), e.g., in a subject in need thereof.

Compounds

Compounds described herein reduce the level of an activity related to BRD9, or a related downstream effect, or reduce the level of BRD9 in a cell or subject. Exemplary compounds described herein have the structure according to Formula I or Formula II.

Formula I is:

embedded image

where

R¹is H, optionally substituted C₁-C₆alkyl, optionally substituted C₂-C₆alkenyl, optionally substituted C₁-C₆heteroalkyl, or optionally substituted C₃-C₁₀carbocyclyl;

Z¹is CR²or N;

embedded image

X¹is CR^X1or N;

X²is O or S;

R^X1is H or optionally substituted C₁-C₆alkyl;

Formula II is:

A-L-B Formula II,

where

L is a linker;

B is a degradation moiety; and

A has the structure of Formula III:

embedded image

where

R¹is, independently, H, optionally substituted C₁-C₆alkyl, optionally substituted C₂-C₆alkenyl, optionally substituted C₁-C₆heteroalkyl, or optionally substituted C₃-C₁₀carbocyclyl;

Z¹is CR²or N;

embedded image

X¹is CR^X1or N;

X²is O or S;

R^X1is H or optionally substituted C₁-C₆alkyl;

R³is H,

embedded image

R³′ is absent, optionally substituted C₁-C₆alkylene, optionally substituted C₂-C₉heteroarylene, or optionally substituted C₁-C₆heteroalkylene;

G is

embedded image

optionally substituted C₃-C₁₀carbocyclyl, C₂-C₉heterocyclyl, optionally substituted C₆-C₁₀aryl, or optionally substituted C₂-C₉heteroaryl;

G′ is optionally substituted C₃-C₁₀carbocyclylene, C₂-C₉heterocyclylene, optionally substituted C₆-C₁₀arylene, or optionally substituted C₂-C₉heteroarylene; and

A¹is a bond between A and the linker,

where G is

embedded image

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound has the structure of any one of compounds B1-B10 in Table 1, or a pharmaceutically acceptable salt thereof.

Other embodiments, as well as exemplary methods for the synthesis of production of these compounds, are described herein.

Pharmaceutical Uses

The compounds described herein are useful in the methods of the invention and, while not bound by theory, are believed to exert their desirable effects through their ability to modulate the level, status, and/or activity of a BAF complex, e.g., by inhibiting the activity or level of the BRD9 protein in a cell within the BAF complex in a mammal.

An aspect of the present invention relates to methods of treating disorders related to BRD9 such as cancer in a subject in need thereof. In some embodiments, the compound is administered in an amount and for a time effective to result in one of (or more, e.g., two or more, three or more, four or more of): (a) reduced tumor size, (b) reduced rate of tumor growth, (c) increased tumor cell death (d) reduced tumor progression, (e) reduced number of metastases, (f) reduced rate of metastasis, (g) decreased tumor recurrence (h) increased survival of subject, and (i) increased progression free survival of a subject.

Treating cancer can result in a reduction in size or volume of a tumor. For example, after treatment, tumor size is reduced by 5% or greater (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or greater) relative to its size prior to treatment. Size of a tumor may be measured by any reproducible means of measurement. For example, the size of a tumor may be measured as a diameter of the tumor.

Treating cancer may further result in a decrease in number of tumors. For example, after treatment, tumor number is reduced by 5% or greater (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or greater) relative to number prior to treatment. Number of tumors may be measured by any reproducible means of measurement, e.g., the number of tumors may be measured by counting tumors visible to the naked eye or at a specified magnification (e.g., 2×, 3×, 4×, 5×, 10×, or 50×).

Treating cancer can result in a decrease in number of metastatic nodules in other tissues or organs distant from the primary tumor site. For example, after treatment, the number of metastatic nodules is reduced by 5% or greater (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater) relative to number prior to treatment. The number of metastatic nodules may be measured by any reproducible means of measurement. For example, the number of metastatic nodules may be measured by counting metastatic nodules visible to the naked eye or at a specified magnification (e.g., 2×, 10×, or 50×).

Treating cancer can result in an increase in average survival time of a population of subjects treated according to the present invention in comparison to a population of untreated subjects. For example, the average survival time is increased by more than 30 days (more than 60 days, 90 days, or 120 days). An increase in average survival time of a population may be measured by any reproducible means. An increase in average survival time of a population may be measured, for example, by calculating for a population the average length of survival following initiation of treatment with the compound described herein. An increase in average survival time of a population may also be measured, for example, by calculating for a population the average length of survival following completion of a first round of treatment with a pharmaceutically acceptable salt of a compound described herein.

Treating cancer can also result in a decrease in the mortality rate of a population of treated subjects in comparison to an untreated population. For example, the mortality rate is decreased by more than 2% (e.g., more than 5%, 10%, or 25%). A decrease in the mortality rate of a population of treated subjects may be measured by any reproducible means, for example, by calculating for a population the average number of disease-related deaths per unit time following initiation of treatment with a pharmaceutically acceptable salt of a compound described herein. A decrease in the mortality rate of a population may also be measured, for example, by calculating for a population the average number of disease-related deaths per unit time following completion of a first round of treatment with a pharmaceutically acceptable salt of a compound described herein.

Combination Therapies

A method of the invention can be used alone or in combination with an additional therapeutic agent, e.g., other agents that treat cancer or symptoms associated therewith, or in combination with other types of therapies to treat cancer. In combination treatments, the dosages of one or more of the therapeutic compounds may be reduced from standard dosages when administered alone. For example, doses may be determined empirically from drug combinations and permutations or may be deduced by isobolographic analysis (e.g., Black et al., Neurology 65:S3-S6 (2005)). In this case, dosages of the compounds when combined should provide a therapeutic effect.

In some embodiments, the second therapeutic agent is a chemotherapeutic agent (e.g., a cytotoxic agent or other chemical compound useful in the treatment of 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 urea, methyl hydrazine derivatives, adrenocortical suppressant, adrenocorticosteroides, progestins, estrogens, antiestrogen, androgens, antiandrogen, and gonadotropin-releasing hormone analog. Also included is 5-fluorouracil (5-FU), leucovorin (LV), irenotecan, oxaliplatin, capecitabine, paclitaxel, and doxetaxel. Non-limiting examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclosphosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, trietylenephosphoramide, triethiylenethiophosphoramide and trimethylolomelamine; acetogenins (especially bullatacin and bullatacinone); a camptothecin (including the synthetic analogue topotecan); bryostatin; callystatin; CC-1065 (including its adozelesin, carzelesin and bizelesin synthetic analogues); cryptophycins (particularly cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including the synthetic analogues, KW-2189 and CB1-TM1); eleutherobin; pancratistatin; a sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine; antibiotics such as the enediyne antibiotics (e.g., calicheamicin, especially calicheamicin gammalI and calicheamicin omegalI (see, e.g., Agnew, Chem. Intl. Ed Engl. 33:183-186 (1994)); dynemicin, including dynemicin A; bisphosphonates, such as clodronate; an esperamicin; as well as neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromophores), aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, ADRIAMYCIN® (doxorubicin, including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; anti-adrenals such as aminoglutethimide, mitotane, trilostane; folic acid replenisher such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elformithine; elliptinium acetate; an epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK® polysaccharide complex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin; sizofuran; spirogermanium; tenuazonic acid; triaziquone; 2,2′,2″-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A and anguidine); urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside (“Ara-C”); cyclophosphamide; thiotepa; taxoids, e.g., TAXOL® (paclitaxel; Bristol-Myers Squibb Oncology, Princeton, N.J.), ABRAXANE®, cremophor-free, albumin-engineered nanoparticle formulation of paclitaxel (American Pharmaceutical Partners, Schaumberg, Ill.), and TAXOTERE® doxetaxel (Rhone-Poulenc Rorer, Antony, France); chlorambucil; GEMZAR® gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum coordination complexes such as cisplatin, oxaliplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; NAVELBINE® vinorelbine; novantrone; teniposide; edatrexate; daunomycin; aminopterin; xeloda; ibandronate; irinotecan (e.g., CPT-11); topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid; capecitabine; and pharmaceutically acceptable salts, acids or derivatives of any of the above. Two or more chemotherapeutic agents can be used in a cocktail to be administered in combination with the first therapeutic agent described herein. Suitable dosing regimens of combination chemotherapies are known in the art and described in, for example, Saltz et al., Proc. Am. Soc. Clin. Oncol. 18:233a (1999), and Douillard et al., Lancet 355(9209):1041-1047 (2000).

In some embodiments, the second therapeutic agent is a therapeutic agent which is a biologic such a cytokine (e.g., interferon or an interleukin (e.g., IL-2)) used in cancer treatment. In some embodiments the biologic is an anti-angiogenic agent, such as an anti-VEGF agent, e.g., bevacizumab (AVASTIN®). In some embodiments the biologic is an immunoglobulin-based biologic, e.g., 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); SIMULECT® (basiliximab); SYNAGIS® (palivizumab); REMICADE® (infliximab); HERCEPTIN® (trastuzumab); MYLOTARG® (gemtuzumab ozogamicin); CAM PATH® (alemtuzumab); ZEVALIN® (ibritumomab tiuxetan); HUMIRA® (adalimumab); XOLAIR® (omalizumab); BEXXAR® (tositumomab-l-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® (raxibacumab); BENLYSTA® (belimumab); YERVOY® (ipilimumab); ADCETRIS® (brentuximab vedotin); PERJETA® (pertuzumab); KADCYLA® (ado-trastuzumab emtansine); and GAZYVA® (obinutuzumab). Also included are antibody-drug conjugates.

The second agent may be a therapeutic agent which is a non-drug treatment. For example, the second therapeutic agent is radiation therapy, cryotherapy, hyperthermia, and/or surgical excision of tumor tissue.

The second agent may be a checkpoint inhibitor. In one embodiment, the inhibitor of checkpoint is an inhibitory antibody (e.g., a monospecific antibody such as a monoclonal antibody). The antibody may be, e.g., humanized or fully human. In some embodiments, the inhibitor of checkpoint is a fusion protein, e.g., an Fc-receptor fusion protein. In some embodiments, the inhibitor of checkpoint is an agent, such as an antibody, that interacts with a checkpoint protein. In some embodiments, the inhibitor of checkpoint is an agent, such as an antibody, that interacts with the ligand of a checkpoint protein. In some embodiments, the inhibitor of checkpoint is an inhibitor (e.g., an inhibitory antibody or small molecule inhibitor) of CTLA-4 (e.g., an anti-CTLA4 antibody or fusion a protein such as ipilimumab/YERVOY® or tremelimumab). In some embodiments, the inhibitor of checkpoint is an inhibitor (e.g., an inhibitory antibody or small molecule inhibitor) of PD-1 (e.g., nivolumab/OPDIVO®; pembrolizumab/KEYTRUDA®; pidilizumab/CT-011). In some embodiments, the inhibitor of checkpoint is an inhibitor (e.g., an inhibitory antibody or small molecule inhibitor) of PDL1 (e.g., MPDL3280A/RG7446; MEDI4736; MSB0010718C; BMS 936559). In some embodiments, the inhibitor of checkpoint 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 some embodiments, the inhibitor of checkpoint 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 ligands, or a combination thereof.

In some embodiments, the anti-cancer therapy is a T cell adoptive transfer (ACT) therapy. In some embodiments, the T cell is an activated T cell. The T cell may be modified to express a chimeric antigen receptor (CAR). CAR modified T (CAR-T) cells can be generated by any method known in the art. For example, the CAR-T cells can be generated by introducing a suitable expression vector encoding the CAR to a T cell. Prior to expansion and genetic modification of the T cells, a source of T cells is obtained from a subject. T cells can be obtained from a number of sources, including peripheral blood mononuclear cells, bone marrow, lymph node tissue, cord blood, thymus tissue, tissue from a site of infection, ascites, pleural effusion, spleen tissue, and tumors. In certain embodiments of the present invention, any number of T cell lines available in the art, may be used. In some embodiments, the T cell is an autologous T cell. Whether prior to or after genetic modification of the T cells to express a desirable protein (e.g., a CAR), the T cells can be activated and expanded generally using methods as described, for example, in U.S. Pat. Nos. 6,352,694; 6,534,055; 6,905,680; 6,692,964; 5,858,358; 6,887,466; 6,905,681; 7,144,575; 7,067,318; 7,172,869; 7,232,566; 7,175,843; 5,883,223; 6,905,874; 6,797,514; 6,867,041; and U.S. Patent Application Publication No. 20060121005.

In any of the combination embodiments described herein, the first and second therapeutic agents are administered simultaneously or sequentially, in either order. The first therapeutic agent may 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 hours 16, up to 17 hours, up 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.

Pharmaceutical Compositions

The pharmaceutical compositions described herein are preferably formulated into pharmaceutical compositions for administration to human subjects in a biologically compatible form suitable for administration in vivo.

The compounds described herein may be used in the form of the free base, in the form of salts, solvates, and as prodrugs. All forms are within the methods described herein. In accordance with the methods of the invention, the described compounds or salts, solvates, or prodrugs thereof may be administered to a patient in a variety of forms depending on the selected route of administration, as will be understood by those skilled in the art. The compounds described herein may be administered, for example, by oral, parenteral, buccal, sublingual, nasal, rectal, patch, pump, intratumoral, or transdermal administration and the pharmaceutical compositions formulated accordingly. Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary, intrathecal, rectal, and topical modes of administration. Parenteral administration may be by continuous infusion over a selected period of time.

A compound described herein may be orally administered, for example, with an inert diluent or with an assimilable edible carrier, or it may be enclosed in hard or soft shell gelatin capsules, or it may be compressed into tablets, or it may be incorporated directly with the food of the diet. For oral therapeutic administration, a compound described herein may be incorporated with an excipient and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, and wafers. A compound described herein may also be administered parenterally. Solutions of a compound described herein can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, DMSO, and mixtures thereof with or without alcohol, and in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms. Conventional procedures and ingredients for the selection and preparation of suitable formulations are described, for example, in Remington's Pharmaceutical Sciences (2012, 22nd ed.) and in The United States Pharmacopeia: The National Formulary (USP 41 NF36), published in 2018. The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases the form must be sterile and must be fluid to the extent that may be easily administered via syringe. Compositions for nasal administration may conveniently be formulated as aerosols, drops, gels, and powders. Aerosol formulations typically include a solution or fine suspension of the active substance in a physiologically acceptable aqueous or non-aqueous solvent and are usually presented in single or multidose quantities in sterile form in a sealed container, which can take the form of a cartridge or refill for use with an atomizing device. Alternatively, the sealed container may be a unitary dispensing device, such as a single dose nasal inhaler or an aerosol dispenser fitted with a metering valve which is intended for disposal after use. Where the dosage form includes an aerosol dispenser, it will contain a propellant, which can be a compressed gas, such as compressed air or an organic propellant, such as fluorochlorohydrocarbon. The aerosol dosage forms can also take the form of a pump-atomizer. Compositions suitable for buccal or sublingual administration include tablets, lozenges, and pastilles, where the active ingredient is formulated with a carrier, such as sugar, acacia, tragacanth, gelatin, and glycerine. Compositions for rectal administration are conveniently in the form of suppositories containing a conventional suppository base, such as cocoa butter. A compound described herein may be administered intratumorally, for example, as an intratumoral injection. Intratumoral injection is injection directly into the tumor vasculature and is specifically contemplated for discrete, solid, accessible tumors. Local, regional, or systemic administration also may be appropriate. A compound described herein may advantageously be contacted by administering an injection or multiple injections to the tumor, spaced for example, at approximately, 1 cm intervals. In the case of surgical intervention, the present invention may be used preoperatively, such as to render an inoperable tumor subject to resection. Continuous administration also may be applied where appropriate, for example, by implanting a catheter into a tumor or into tumor vasculature.

The compounds described herein may be administered to an animal, e.g., a human, alone or in combination with pharmaceutically acceptable carriers, as noted herein, the proportion of which is determined by the solubility and chemical nature of the compound, chosen route of administration, and standard pharmaceutical practice.

Dosages

The dosage of the compounds described herein, and/or compositions including a compound described herein, can 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 the symptoms; the frequency of the treatment, and the type of concurrent treatment, if any; and the clearance rate of the compound in the animal to be treated. One of skill in the art can determine the appropriate dosage based on the above factors. The compounds described herein may be administered initially in a suitable dosage that may be adjusted as required, depending on the clinical response. In general, satisfactory results may be obtained when the compounds described herein are administered to a human at a daily dosage of, for example, between 0.05 mg and 3000 mg (measured as the solid form). Dose ranges include, for example, between 10-1000 mg (e.g., 50-800 mg). In some embodiments, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or 1000 mg of the compound is administered.

Alternatively, the dosage amount can be calculated using the body weight of the patient. For example, the dose of a compound, or pharmaceutical composition thereof, administered to a patient may range from 0.1-100 mg/kg (e.g., 0.1-50 mg/kg (e.g., 0.25-25 mg/kg)). In exemplary, non-limiting embodiments, the dose may range from 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 from 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).

Kits

The invention also features kits including (a) a pharmaceutical composition including an agent that reduces the level and/or activity of BRD9 in a cell or subject described herein, and (b) a package insert with instructions to perform any of the methods described herein. In some embodiments, the kit includes (a) a pharmaceutical composition including an agent that reduces the level and/or activity of BRD9 in a cell or subject described herein, (b) an additional therapeutic agent (e.g., an anti-cancer agent), and (c) a package insert with instructions to perform any of the methods described herein.

EXAMPLES
Example 1—High Density Tiling sgRNA Screen Against Human BAF Complex Subunits in Synovial Sarcoma Cell Line SYO1

The following example shows that BRD9 sgRNA inhibits cell growth in synovial sarcoma cells.

Procedure: To perform high density sgRNA tiling screen, an sgRNA library against BAF complex subunits was custom synthesized at Cellecta (Mountain View, Calif.). Sequences of DNA encoding the BRD9-targeting sgRNAs used in this screen are listed in Table 3. Negative and positive control sgRNA were included in the library. Negative controls consisted of 200 sgRNAs that do not target human genome. The positive controls are sgRNAs targeting essential genes (CDC16, GTF2B, HSPA5, HSPA9, PAFAH1B1, PCNA, POLR2L, RPL9, and SF3A3). DNA sequences encoding all positive and negative control sgRNAs are listed in Table 4. Procedures for virus production, cell infection, and performing the sgRNA screen were previously described (Tsherniak et al, Cell 170:564-576 (2017); Munoz et al, Cancer Discovery 6:900-913 (2016)). For each sgRNA, 50 counts were added to the sequencing counts and for each time point the resulting counts were normalized to the total number of counts. The log 2 of the ratio between the counts (defined as dropout ratio) at day 24 and day 1 post-infection was calculated. For negative control sgRNAs, the 2.5 and 97.5 percentile of the log 2 dropout ratio of all non-targeting sgRNAs was calculated and considered as background (grey box in the graph). Protein domains were obtained from PFAM regions defined for the UNIPROT identifier: Q9H8M2.

Results: As shown in FIG. 1, targeted inhibition of the GBAF complex component BRD9 by sgRNA resulted in growth inhibition of the SYO1 synovial sarcoma cell line. sgRNAs against other components of the BAF complexes resulted in increased proliferation of cells, inhibition of cell growth, or had no effect on SYO1 cells. These data show that targeting various subunits of the GBAF complex represents a therapeutic strategy for the treatment of synovial sarcoma.

TABLE 3

BRD9 sgRNA Library

SEQ ID

NO
Nucleic Acid Sequence

203
CAAGAAGCACAAGAAGCACA

204
CTTGTGCTTCTTGCCCATGG

205
CTTCTTGTGCTTCTTGCCCA

206
ACAAGAAGCACAAGGCCGAG

207
CTCGTAGGACGAGCGCCACT

208
CGAGTGGCGCTCGTCCTACG

209
GAGTGGCGCTCGTCCTACGA

210
AGGCTTCTCCAGGGGCTTGT

211
AGATTATGCCGACAAGCCCC

212
ACCTTCAGGACTAGCTTTAG

213
AGCTTTAGAGGCTTCTCCAG

214
CTAGCTTTAGAGGCTTCTCC

215
TAGCTTTAGAGGCTTCTCCA

216
CTAAAGCTAGTCCTGAAGGT

217
GCCTCTAAAGCTAGTCCTGA

218
CTTCACTTCCTCCGACCTTC

219
AAGCTAGTCCTGAAGGTCGG

220
AGTGAAGTGACTGAACTCTC

221
GTGACTGAACTCTCAGGATC

222
ATAGTAACTGGAGTCGTGGC

223
CATCATAGTAACTGGAGTCG

224
TGACCTGTCATCATAGTAAC

225
ACTCCAGTTACTATGATGAC

226
CTTTGTGCCTCTCTCGCTCA

227
GGTCAGACCATGAGCGAGAG

228
GAAGAAGAAGAAGTCCGAGA

229
GTCCAGATGCTTCTCCTTCT

230
GTCCGAGAAGGAGAAGCATC

231
GGAGAAGCATCTGGACGATG

232
TGAGGAAAGAAGGAAGCGAA

233
ATCTGGACGATGAGGAAAGA

234
AGAAGAAGCGGAAGCGAGAG

235
GAAGAAGCGGAAGCGAGAGA

236
CCGCCCAGGAAGAGAAGAAG

237
AGAGAGGGAGCACTGTGACA

238
AGGGAGCACTGTGACACGGA

239
GAGGGAGCACTGTGACACGG

240
GCACTGTGACACGGAGGGAG

241
GAGGCTGACGACTTTGATCC

242
AGGCTGACGACTTTGATCCT

243
TCCACCTCCACCTTCTTCCC

244
CGACTTTGATCCTGGGAAGA

245
CTTTGATCCTGGGAAGAAGG

246
TGATCCTGGGAAGAAGGTGG

247
TCCTGGGAAGAAGGTGGAGG

248
CGGACTGGCCGATCTGGGGG

249
ACGCTCGGACTGGCCGATCT

250
AGGTGGAGCCGCCCCCAGAT

251
CGCTCGGACTGGCCGATCTG

252
GCTCGGACTGGCCGATCTGG

253
CACGCTCGGACTGGCCGATC

254
TGTGTCCGGCACGCTCGGAC

255
CTGGCTGTGTCCGGCACGCT

256
ATCGGCCAGTCCGAGCGTGC

257
CACCCTTGCCTGGCTGTGTC

258
CGAGCGTGCCGGACACAGCC

259
TGTTCCAGGAGTTGCTGAAT

260
CACACCTATTCAGCAACTCC

261
GCTGGCGGAGGAAGTGTTCC

262
TTTACCTCTGAAGCTGGCGG

263
CCCCGGTTTACCTCTGAAGC

264
ACTTCCTCCGCCAGCTTCAG

265
CAGGAAAAGCAAAAAATCCA

266
GCTTTCAGAAAAGATCCCCA

267
AGGAAAAGCAAAAAATCCAT

268
GGAAAAGCAAAAAATCCATG

269
GGAGCAATTGCATCCGTGAC

270
GTCACGGATGCAATTGCTCC

271
TTTATTATCATTGAATATCC

272
AATGATAATAAAACATCCCA

273
ATAAAACATCCCATGGATTT

274
TTCATGGTGCCAAAATCCAT

275
TTTCATGGTGCCAAAATCCA

276
TAATGAATACAAGTCAGTTA

277
CAAGTCAGTTACGGAATTTA

278
ATAATGCAATGACATACAAT

279
AACTTGTAGTACACGGTATC

280
CTTCGCCAACTTGTAGTACA

281
AGATACCGTGTACTACAAGT

282
GCGAAGAAGATCCTTCACGC

283
TCATCTTAAAGCCTGCGTGA

284
TTCTCAGCAGGCAGCTCTTT

285
CAATGAAGATACAGCTGTTG

286
ACTGGTACAACTTCAGGGAC

287
CTTGTACTGGTACAACTTCA

288
ACTTGTACTGGTACAACTTC

289
TTGGCAGTTTCTACTTGTAC

290
TACCTGATAACTTCTCTACT

291
AGCCGAGTAGAGAAGTTATC

292
AGCTGCATGTTTGAGCCTGA

293
GCTGCATGTTTGAGCCTGAA

294
AAGCTGCAGGCATTCCCTTC

295
GGTACTGTCCGTCAAGCTGC

296
AGGGAATGCCTGCAGCTTGA

297
CTTGACGGACAGTACCGCAG

298
CGCCAGCACGTGCTCCTCTG

299
TACCGCAGAGGAGCACGTGC

300
AGAGGAGCACGTGCTGGCGC

301
GGAGCACGTGCTGGCGCTGG

302
AGCACGCAGCTGACGAAGCT

303
GCACGCAGCTGACGAAGCTC

304
CAGCTGACGAAGCTCGGGAC

305
AAGCTCGGGACAGGATCAAC

306
CCTTGCCGCCTGGGAGGAAC

307
AGGATCAACCGGTTCCTCCC

308
ATCAACCGGTTCCTCCCAGG

309
GCACTACCTTGCCGCCTGGG

310
AGAGCACTACCTTGCCGCCT

311
CCGGTTCCTCCCAGGCGGCA

312
TCCTCTTCAGATAGCCCATC

313
ATGGGCTATCTGAAGAGGAA

314
GGGCTATCTGAAGAGGAACG

315
TGGGCTATCTGAAGAGGAAC

316
TATCTGAAGAGGAACGGGGA

317
ATCTGAAGAGGAACGGGGAC

318
TGTTGACCACGCTGTAGAGC

319
GCTCTACAGCGTGGTCAACA

320
CGGGAGCCTGCTCTACAGCG

321
CGTGGTCAACACGGCCGAGC

322
CCCACCATCAGCGTCCGGCT

323
ACGGCCGAGCCGGACGCTGA

324
GGGCACCCACCATCAGCGTC

325
GCCGAGCCGGACGCTGATGG

326
CCATGTCCGTGTTGCAGAGG

327
CCGAGCCGGACGCTGATGGT

328
CGAGCTCAAGTCCACCGGGT

329
GCGAGCTCAAGTCCACCGGG

330
AGAGCGAGCTCAAGTCCACC

331
GAGAGCGAGCTCAAGTCCAC

332
GAAGCCTGGGAGTAGCTTAC

333
CTCTCCAGTAAGCTACTCCC

334
AGCCCAGCGTGGTGAAGCCT

335
AAGCCCAGCGTGGTGAAGCC

336
ACTCCCAGGCTTCACCACGC

337
CTCCCAGGCTTCACCACGCT

338
CTCGTCTTTGAAGCCCAGCG

339
CACTGGAGAGAAAGGTGACT

340
GCACTGGAGAGAAAGGTGAC

341
AGTAGTGGCACTGGAGAGAA

342
CGAAAGCGCAGTAGTGGCAC

343
CTGCATCGAAAGCGCAGTAG

344
ATGCAGAATAATTCAGTATT

345
AGTATTTGGCGACTTGAAGT

346
CGACTTGAAGTCGGACGAGA

347
GAGCTGCTCTACTCAGCCTA

348
CACGCCTGTCTCATCTCCGT

349
TCAGCCTACGGAGATGAGAC

350
CAGGCGTGCAGTGTGCGCTG

351
CCGCGGCCCCTCTAGCCTGC

352
CATCCTTCACAAACTCCTGC

353
TAGCCTGCAGGAGTTTGTGA

354
CAGGAGTTTGTGAAGGATGC

355
AGGAGTTTGTGAAGGATGCT

356
TGGGAGCTACAGCAAGAAAG

357
GAGCTACAGCAAGAAAGTGG

358
GAAAGTGGTGGACGACCTCC

359
CGCCTGTGATCTGGTCCAGG

360
CTCCGCCTGTGATCTGGTCC

361
GACCTCCTGGACCAGATCAC

362
CTCCTGGACCAGATCACAGG

363
GCTGGAAGAGCGTCCTAGAG

364
TGCAGCCCACCTGCTTCAGC

365
GACGCTCTTCCAGCTGAAGC

366
CTCTTCCAGCTGAAGCAGGT

367
GCTCTTCCAGCTGAAGCAGG

368
CCTCCAGATGAAGCCAAGGT

369
GCTTCATCTGGAGGCTTCAT

370
GGCTTCATCTGGAGGCTTCA

371
CTTACCTTGGCTTCATCTGG

372
AAACTTACCTTGGCTTCATC

373
GAAGCCTCCAGATGAAGCCA

374
TCCTAGGGTGTCCCCAACCT

375
CCTAGGGTGTCCCCAACCTG

376
GTGTCTGTCTCCACAGGTTG

377
TGTGTCTGTCTCCACAGGTT

378
CCACAGGTTGGGGACACCCT

379
AGAGCTGCTGCTGTCTCCTA

380
CAGAGCTGCTGCTGTCTCCT

381
AGACAGCAGCAGCTCTGTTC

382
ATCCACAGAAACGTCGGGAT

383
GAGATATCCACAGAAACGTC

384
GGAGATATCCACAGAAACGT

385
GTCCTATCCCGACGTTTCTG

386
TCTCCATGCTCAGCTCTCTG

387
CTCACCCAGAGAGCTGAGCA

388
ATCTCCATGCTCAGCTCTCT

389
TATCTCCATGCTCAGCTCTC

390
ATGTCCTGTTTACACAGGGA

391
TTACACAGGGAAGGTGAAGA

392
AGTTCAAATGGCTGTCGTCA

393
TGACGACAGCCATTTGAACT

394
AAGTTCAAATGGCTGTCGTC

395
TCGTCTCATCCAAGTTCAAA

396
TGAGACGACGAAGCTCCTGC

397
GTGCTTCGTGCAGGTCCTGC

398
GCAGGACCTGCACGAAGCAC

399
GCTCCGCCTGTGCTTCGTGC

400
GGACCTGCACGAAGCACAGG

401
CACGAAGCACAGGCGGAGCG

402
AGGCGGAGCGCGGCGGCTCT

403
AGGGAGCTGAGGTTGGACGA

404
GTTGGACAGGGAGCTGAGGT

405
AGGCGTTGGACAGGGAGCTG

406
CCCTCTCGGAGGCGTTGGAC

407
CCTCTCGGAGGCGTTGGACA

408
CTGGTCCCTCTCGGAGGCGT

409
CCCTGTCCAACGCCTCCGAG

410
CCTGTCCAACGCCTCCGAGA

411
GTGGTGCTGGTCCCTCTCGG

412
CAGGTGGTGCTGGTCCCTCT

413
GCATCTCACCCAGGTGGTGC

414
CGAGAGGGACCAGCACCACC

415
GAGAGGGACCAGCACCACCT

416
GTGGGGGCATCTCACCCAGG

417
CCCCGACACTCAGGCGAGAA

418
TCCCCGACACTCAGGCGAGA

419
AGCCCTTCTCGCCTGAGTGT

420
CTGGCTGCTCCCCGACACTC

421
CCCTTCTCGCCTGAGTGTCG

422
GCCCTTCTCGCCTGAGTGTC

423
TAGGGGTCGTGGGTGACGTC

424
AAGAAACTCATAGGGGTCGT

425
GAAGAAACTCATAGGGGTCG

426
GAGACTGAAGAAACTCATAG

427
GGAGACTGAAGAAACTCATA

428
TGGAGACTGAAGAAACTCAT

429
TCTTCAGTCTCCAGAGCCTG

430
TTGGCAGAGGCCGCAGGCTC

431
TAGGTCTTGGCAGAGGCCGC

432
CTAGAGTTAGGTCTTGGCAG

433
GGTGGTCTAGAGTTAGGTCT

TABLE 4

Control sgRNA Library

SEQ

ID

NO.
gRNA Label
Gene
Nucleic Acid Sequence

434
1|sg_Non_Targeting_Human_0001|
Non_Targeting_Human
GTAGCGAACGTGTCCGGCGT

Non_Targeting_Human

435
1|sg_Non_Targeting_Human_0002|
Non_Targeting_Human
GACCGGAACGATCTCGCGTA

Non_Targeting_Human

436
1|sg_Non_Targeting_Human_0003|
Non_Targeting_Human
GGCAGTCGTTCGGTTGATAT

Non_Targeting_Human

437
1|sg_Non_Targeting_Human_0004|
Non_Targeting_Human
GCTTGAGCACATACGCGAAT

Non_Targeting_Human

438
1|sg_Non_Targeting_Human_0005|
Non_Targeting_Human
GTGGTAGAATAACGTATTAC

Non_Targeting_Human

439
1|sg_Non_Targeting_Human_0006|
Non_Targeting_Human
GTCATACATGGATAAGGCTA

Non_Targeting_Human

440
1|sg_Non_Targeting_Human_0007|
Non_Targeting_Human
GATACACGAAGCATCACTAG

Non_Targeting_Human

441
1|sg_Non_Targeting_Human_0008|
Non_Targeting_Human
GAACGTTGGCACTACTTCAC

Non_Targeting_Human

442
1|sg_Non_Targeting_Human_0009|
Non_Targeting_Human
GATCCATGTAATGCGTTCGA

Non_Targeting_Human

443
1|sg_Non_Targeting_Human_0010|
Non_Targeting_Human
GTCGTGAAGTGCATTCGATC

Non_Targeting_Human

444
1|sg_Non_Targeting_Human_0011|
Non_Targeting_Human
GTTCGACTCGCGTGACCGTA

Non_Targeting_Human

445
1|sg_Non_Targeting_Human_0012|
Non_Targeting_Human
GAATCTACCGCAGCGGTTCG

Non_Targeting_Human

446
1|sg_Non_Targeting_Human_0013|
Non_Targeting_Human
GAAGTGACGTCGATTCGATA

Non_Targeting_Human

447
1|sg_Non_Targeting_Human_0014|
Non_Targeting_Human
GCGGTGTATGACAACCGCCG

Non_Targeting_Human

448
1|sg_Non_Targeting_Human_0015|
Non_Targeting_Human
GTACCGCGCCTGAAGTTCGC

Non_Targeting_Human

449
1|sg_Non_Targeting_Human_0016|
Non_Targeting_Human
GCAGCTCGTGTGTCGTACTC

Non_Targeting_Human

450
1|sg_Non_Targeting_Human_0017|
Non_Targeting_Human
GCGCCTTAAGAGTACTCATC

Non_Targeting_Human

451
1|sg_Non_Targeting_Human_0018|
Non_Targeting_Human
GAGTGTCGTCGTTGCTCCTA

Non_Targeting_Human

452
1|sg_Non_Targeting_Human_0019|
Non_Targeting_Human
GCAGCTCGACCTCAAGCCGT

Non_Targeting_Human

453
1|sg_Non_Targeting_Human_0020|
Non_Targeting_Human
GTATCCTGACCTACGCGCTG

Non_Targeting_Human

454
1|sg_Non_Targeting_Human_0021|
Non_Targeting Human
GTGTATCTCAGCACGCTAAC

Non_Targeting_Human

455
1|sg_Non_Targeting_Human_0022|
Non_Targeting_Human
GTCGTCATACAACGGCAACG

Non_Targeting_Human

456
1|sg_Non_Targeting_Human_0023|
Non_Targeting_Human
GTCGTGCGCTTCCGGCGGTA

Non_Targeting_Human

457
1|sg_Non_Targeting_Human_0024|
Non_Targeting_Human
GCGGTCCTCAGTAAGCGCGT

Non_Targeting_Human

458
1|sg_Non_Targeting_Human_0025|
Non_Targeting_Human
GCTCTGCTGCGGAAGGATTC

Non_Targeting_Human

459
1|sg_Non_Targeting_Human_0026|
Non_Targeting_Human
GCATGGAGGAGCGTCGCAGA

Non_Targeting_Human

460
1|sg_Non_Targeting_Human_0027|
Non_Targeting_Human
GTAGCGCGCGTAGGAGTGGC

Non_Targeting_Human

461
1|sg_Non_Targeting_Human_0028|
Non_Targeting_Human
GATCACCTGCATTCGTACAC

Non_Targeting_Human

462
1|sg_Non_Targeting_Human_0029|
Non_Targeting_Human
GCACACCTAGATATCGAATG

Non_Targeting_Human

463
1|sg_Non_Targeting_Human_0030|
Non_Targeting_Human
GTTGATCAACGCGCTTCGCG

Non_Targeting_Human

464
1|sg_Non_Targeting_Human_0031|
Non_Targeting_Human
GCGTCTCACTCACTCCATCG

Non_Targeting_Human

465
1|sg_Non_Targeting_Human_0032|
Non_Targeting_Human
GCCGACCAACGTCAGCGGTA

Non_Targeting_Human

466
1|sg_Non_Targeting_Human_0033|
Non_Targeting_Human
GGATACGGTGCGTCAATCTA

Non_Targeting_Human

467
1|sg_Non_Targeting_Human_0034|
Non_Targeting_Human
GAATCCAGTGGCGGCGACAA

Non_Targeting_Human

468
1|sg_Non_Targeting_Human_0035|
Non_Targeting_Human
GCACTGTCAGTGCAACGATA

Non_Targeting_Human

469
1|sg_Non_Targeting_Human_0036|
Non_Targeting_Human
GCGATCCTCAAGTATGCTCA

Non_Targeting_Human

470
1|sg_Non_Targeting_Human_0037|
Non_Targeting_Human
GCTAATATCGACACGGCCGC

Non_Targeting_Human

471
1|sg_Non_Targeting_Human_0038|
Non_Targeting_Human
GGAGATGCATCGAAGTCGAT

Non_Targeting_Human

472
1|sg_Non_Targeting_Human_0039|
Non_Targeting_Human
GGATGCACTCCATCTCGTCT

Non_Targeting_Human

473
1|sg_Non_Targeting_Human_0040|
Non_Targeting_Human
GTGCCGAGTAATAACGCGAG

Non_Targeting_Human

474
1|sg_Non_Targeting_Human_0041|
Non_Targeting_Human
GAGATTCCGATGTAACGTAC

Non_Targeting_Human

475
1|sg_Non_Targeting_Human_0042|
Non_Targeting_Human
GTCGTCACGAGCAGGATTGC

Non_Targeting_Human

476
1|sg_Non_Targeting_Human_0043|
Non_Targeting_Human
GCGTTAGTCACTTAGCTCGA

Non_Targeting_Human

477
1|sg_Non_Targeting_Human_0044|
Non_Targeting_Human
GTTCACACGGTGTCGGATAG

Non_Targeting_Human

478
1|sg_Non_Targeting_Human_0045|
Non_Targeting_Human
GGATAGGTGACCTTAGTACG

Non_Targeting_Human

479
1|sg_Non_Targeting_Human_0046|
Non_Targeting_Human
GTATGAGTCAAGCTAATGCG

Non_Targeting_Human

480
1|sg_Non_Targeting_Human_0047|
Non_Targeting_Human
GCAACTATTGGAATACGTGA

Non_Targeting_Human

481
1|sg_Non_Targeting_Human_0048|
Non_Targeting_Human
GTTACCTTCGCTCGTCTATA

Non_Targeting_Human

482
1|sg_Non_Targeting_Human_0049|
Non_Targeting_Human
GTACCGAGCACCACAGGCCG

Non_Targeting_Human

483
1|sg_Non_Targeting_Human_0050|
Non_Targeting_Human
GTCAGCCATCGGATAGAGAT

Non_Targeting_Human

484
1|sg_Non_Targeting_Human_0051|
Non_Targeting_Human
GTACGGCACTCCTAGCCGCT

Non_Targeting_Human

485
1|sg_Non_Targeting_Human_0052|
Non_Targeting_Human
GGTCCTGTCGTATGCTTGCA

Non_Targeting_Human

486
1|sg_Non_Targeting_Human_0053|
Non_Targeting_Human
GCCGCAATATATGCGGTAAG

Non_Targeting_Human

487
1|sg_Non_Targeting_Human_0054|
Non_Targeting_Human
GCGCACGTATAATCCTGCGT

Non_Targeting_Human

488
1|sg_Non_Targeting_Human_0055|
Non_Targeting_Human
GTGCACAACACGATCCACGA

Non_Targeting_Human

489
1|sg_Non_Targeting_Human_0056|
Non_Targeting_Human
GCACAATGTTGACGTAAGTG

Non_Targeting_Human

490
1|sg_Non_Targeting_Human_0057|
Non_Targeting_Human
GTAAGATGCTGCTCACCGTG

Non_Targeting_Human

491
1|sg_Non_Targeting_Human_0058|
Non_Targeting_Human
GTCGGTGATCCAACGTATCG

Non_Targeting_Human

492
1|sg_Non_Targeting_Human_0059|
Non_Targeting_Human
GAGCTAGTAGGACGCAAGAC

Non_Targeting_Human

493
1|sg_Non_Targeting_Human_0060|
Non_Targeting_Human
GTACGTGGAAGCTTGTGGCC

Non_Targeting_Human

494
1|sg_Non_Targeting_Human_0061|
Non_Targeting_Human
GAGAACTGCCAGTTCTCGAT

Non_Targeting_Human

495
1|sg_Non_Targeting_Human_0062|
Non_Targeting_Human
GCCATTCGGCGCGGCACTTC

Non_Targeting_Human

496
1|sg_Non_Targeting_Human_0063|
Non_Targeting_Human
GCACACGACCAATCCGCTTC

Non_Targeting_Human

497
1|sg_Non_Targeting_Human_0064|
Non_Targeting_Human
GAGGTGATCGATTAAGTACA

Non_Targeting_Human

498
1|sg_Non_Targeting_Human_0065|
Non_Targeting_Human
GTCACTCGCAGACGCCTAAC

Non_Targeting_Human

499
1|sg_Non_Targeting_Human_0066|
Non Targeting_Human
GCGCTACGGAATCATACGTT

Non_Targeting_Human

500
1|sg_Non_Targeting_Human_0067|
Non_Targeting_Human
GGTAGGACCTCACGGCGCGC

Non_Targeting_Human

501
1|sg_Non_Targeting_Human_0068|
Non_Targeting_Human
GAACTGCATCTTGTTGTAGT

Non_Targeting_Human

502
1|sg_Non_Targeting_Human_0069|
Non_Targeting_Human
GATCCTGATCCGGCGGCGCG

Non_Targeting_Human

503
1|sg_Non_Targeting_Human_0070|
Non_Targeting_Human
GGTATGCGCGATCCTGAGTT

Non_Targeting_Human

504
1|sg_Non_Targeting_Human_0071|
Non_Targeting_Human
GCGGAGCTAGAGAGCGGTCA

Non_Targeting_Human

505
1|sg_Non_Targeting_Human_0072|
Non_Targeting_Human
GAATGGCAATTACGGCTGAT

Non_Targeting_Human

506
1|sg_Non_Targeting_Human_0073|
Non_Targeting_Human
GTATGGTGAGTAGTCGCTTG

Non_Targeting_Human

507
1|sg_Non_Targeting_Human_0074|
Non_Targeting_Human
GTGTAATTGCGTCTAGTCGG

Non_Targeting_Human

508
1|sg_Non_Targeting_Human_0075|
Non_Targeting_Human
GGTCCTGGCGAGGAGCCTTG

Non_Targeting_Human

509
1|sg_Non_Targeting_Human_0076|
Non_Targeting_Human
GAAGATAAGTCGCTGTCTCG

Non_Targeting_Human

510
1|sg_Non_Targeting_Human_0077|
Non_Targeting_Human
GTCGGCGTTCTGTTGTGACT

Non_Targeting_Human

511
1|sg_Non_Targeting_Human_0078|
Non_Targeting_Human
GAGGCAAGCCGTTAGGTGTA

Non_Targeting_Human

512
1|sg_Non_Targeting_Human_0079|
Non_Targeting_Human
GCGGATCCAGATCTCATTCG

Non_Targeting_Human

513
1|sg_Non_Targeting_Human_0080|
Non_Targeting_Human
GGAACATAGGAGCACGTAGT

Non_Targeting_Human

514
1|sg_Non_Targeting_Human_0081|
Non_Targeting_Human
GTCATCATTATGGCGTAAGG

Non_Targeting_Human

515
1|sg_Non_Targeting_Human_0082|
Non_Targeting_Human
GCGACTAGCGCCATGAGCGG

Non_Targeting_Human

516
1|sg_Non_Targeting_Human_0083|
Non_Targeting_Human
GGCGAAGTTCGACATGACAC

Non_Targeting_Human

517
1|sg_Non_Targeting_Human_0084|
Non_Targeting_Human
GCTGTCGTGTGGAGGCTATG

Non_Targeting_Human

518
1|sg_Non_Targeting_Human_0085|
Non_Targeting_Human
GCGGAGAGCATTGACCTCAT

Non_Targeting_Human

519
1|sg_Non_Targeting_Human_0086|
Non_Targeting_Human
GACTAATGGACCAAGTCAGT

Non_Targeting_Human

520
1|sg_Non_Targeting_Human_0087|
Non_Targeting_Human
GCGGATTAGAGGTAATGCGG

Non_Targeting_Human

521
1|sg_Non_Targeting_Human_0088|
Non_Targeting_Human
GCCGACGGCAATCAGTACGC

Non_Targeting_Human

522
1|sg_Non_Targeting_Human_0089|
Non_Targeting_Human
GTAACCTCTCGAGCGATAGA

Non_Targeting_Human

523
1|sg_Non_Targeting_Human_0090|
Non_Targeting_Human
GACTTGTATGTGGCTTACGG

Non_Targeting_Human

524
1|sg_Non_Targeting_Human_0091|
Non_Targeting_Human
GTCACTGTGGTCGAACATGT

Non_Targeting_Human

525
1|sg_Non_Targeting_Human_0092|
Non_Targeting_Human
GTACTCCAATCCGCGATGAC

Non_Targeting_Human

526
1|sg_Non_Targeting_Human_0093|
Non_Targeting_Human
GCGTTGGCACGATGTTACGG

Non_Targeting_Human

527
1|sg_Non_Targeting_Human_0094|
Non_Targeting_Human
GAACCAGCCGGCTAGTATGA

Non_Targeting_Human

528
1|sg_Non_Targeting_Human_0095|
Non_Targeting_Human
GTATACTAGCTAACCACACG

Non_Targeting_Human

529
1|sg_Non_Targeting_Human_0096|
Non_Targeting_Human
GAATCGGAATAGTTGATTCG

Non_Targeting_Human

530
1|sg_Non_Targeting_Human_0097|
Non_Targeting_Human
GAGCACTTGCATGAGGCGGT

Non_Targeting_Human

531
1|sg_Non_Targeting_Human_0098|
Non Targeting_Human
GAACGGCGATGAAGCCAGCC

Non_Targeting_Human

532
1|sg_Non_Targeting_Human_0099|
Non_Targeting_Human
GCAACCGAGATGAGAGGTTC

Non_Targeting_Human

533
1|sg_Non_Targeting_Human_0100|
Non_Targeting_Human
GCAAGATCAATATGCGTGAT

Non_Targeting_Human

534
1|sg_Non_Targeting_Human_GA_0101|
Non_Targeting_Human
ACGGAGGCTAAGCGTCGCAA

Non_Targeting_Human

535
1|sg_Non_Targeting_Human_GA_0102|
Non_Targeting_Human
CGCTTCCGCGGCCCGTTCAA

Non_Targeting_Human

536
1|sg_Non_Targeting_Human_GA_0103|
Non_Targeting_Human
ATCGTTTCCGCTTAACGGCG

Non_Targeting_Human

537
1|sg_Non_Targeting_Human_GA_0104|
Non_Targeting_Human
GTAGGCGCGCCGCTCTCTAC

Non_Targeting_Human

538
1|sg_Non_Targeting_Human_GA_0105|
Non_Targeting_Human
CCATATCGGGGCGAGACATG

Non_Targeting_Human

539
1|sg_Non_Targeting_Human_GA_0106|
Non_Targeting_Human
TACTAACGCCGCTCCTACAG

Non_Targeting_Human

540
1|sg_Non_Targeting_Human_GA_0107|
Non_Targeting Human
TGAGGATCATGTCGAGCGCC

Non_Targeting_Human

541
1|sg_Non_Targeting_Human_GA_0108|
Non_Targeting Human
GGGCCCGCATAGGATATCGC

Non_Targeting_Human

542
1|sg_Non_Targeting_Human_GA_0109|
Non_Targeting_Human
TAGACAACCGCGGAGAATGC

Non_Targeting_Human

543
1|sg_Non Targeting_Human_GA_0110|
Non_Targeting_Human
ACGGGCGGCTATCGCTGACT

Non_Targeting_Human

544
1|sg_Non_Targeting_Human_GA_0111|
Non_Targeting_Human
CGCGGAAATTTTACCGACGA

Non_Targeting_Human

545
1|sg_Non_Targeting_Human_GA_0112|
Non_Targeting_Human
CTTACAATCGTCGGTCCAAT

Non_Targeting_Human

546
1|sg_Non_Targeting_Human_GA_0113|
Non_Targeting_Human
GCGTGCGTCCCGGGTTACCC

Non_Targeting_Human

547
1|sg_Non_Targeting_Human_GA_0114|
Non_Targeting_Human
CGGAGTAACAAGCGGACGGA

Non_Targeting_Human

548
1|sg_Non_Targeting_Human_GA_0115|
Non_Targeting_Human
CGAGTGTTATACGCACCGTT

Non_Targeting_Human

549
1|sg_Non_Targeting_Human_GA_0116|
Non_Targeting_Human
CGACTAACCGGAAACTTTTT

Non_Targeting_Human

550
1|sg_Non_Targeting_Human_GA_0117|
Non_Targeting_Human
CAACGGGTTCTCCCGGCTAC

Non_Targeting_Human

551
1|sg_Non_Targeting_Human_GA_0118|
Non_Targeting_Human
CAGGAGTCGCCGATACGCGT

Non_Targeting_Human

552
1|sg_Non_Targeting_Human_GA_0119|
Non_Targeting_Human
TTCACGTCGTCTCGCGACCA

Non_Targeting_Human

553
1|sg_Non_Targeting_Human_GA_0120|
Non_Targeting_Human
GTGTCGGATTCCGCCGCTTA

Non_Targeting_Human

554
1|sg_Non_Targeting_Human_GA_0121|
Non_Targeting_Human
CACGAACTCACACCGCGCGA

Non_Targeting_Human

555
1|sg_Non_Targeting_Human_GA_0122|
Non_Targeting_Human
CGCTAGTACGCTCCTCTATA

Non_Targeting_Human

556
1|sg_Non_Targeting_Human_GA_0123|
Non_Targeting_Human
TCGCGCTTGGGTTATACGCT

Non_Targeting_Human

557
1|sg_Non_Targeting_Human_GA_0124|
Non_Targeting_Human
CTATCTCGAGTGGTAATGCG

Non_Targeting_Human

558
1|sg_Non_Targeting_Human_GA_0125|
Non_Targeting_Human
AATCGACTCGAACTTCGTGT

Non_Targeting_Human

559
1|sg_Non_Targeting_Human_GA_0126|
Non Targeting_Human
CCCGATGGACTATACCGAAC

Non_Targeting_Human

560
1|sg_Non Targeting_Human_GA_0127|
Non_Targeting_Human
ACGTTCGAGTACGACCAGCT

Non_Targeting_Human

561
1|sg_Non_Targeting_Human_GA_0128|
Non_Targeting Human
CGCGACGACTCAACCTAGTC

Non_Targeting_Human

562
1|sg_Non_Targeting_Human GA_0129|
Non_Targeting Human
GGTCACCGATCGAGAGCTAG

Non_Targeting_Human

563
1|sg_Non_Targeting_Human_GA_0130|
Non_Targeting_Human
CTCAACCGACCGTATGGTCA

Non_Targeting_Human

564
1|sg_Non_Targeting_Human_GA_0131|
Non_Targeting_Human
CGTATTCGACTCTCAACGCG

Non_Targeting_Human

565
1|sg_Non_Targeting_Human_GA_0132|
Non_Targeting_Human
CTAGCCGCCCAGATCGAGCC

Non_Targeting_Human

566
1|sg_Non_Targeting_Human_GA_0133|
Non_Targeting_Human
GAATCGACCGACACTAATGT

Non_Targeting_Human

567
1|sg_Non_Targeting_Human_GA_0134|
Non Targeting_Human
ACTTCAGTTCGGCGTAGTCA

Non_Targeting_Human

568
1|sg_Non_Targeting_Human_GA_0135|
Non_Targeting_Human
GTGCGATGTCGCTTCAACGT

Non_Targeting_Human

569
1|sg_Non_Targeting_Human_GA_0136|
Non_Targeting_Human
CGCCTAATTTCCGGATCAAT

Non_Targeting Human

570
1|sg_Non_Targeting_Human_GA_0137|
Non_Targeting_Human
CGTGGCCGGAACCGTCATAG

Non_Targeting_Human

571
1|sg_Non_Targeting_Human_GA_0138|
Non_Targeting_Human
ACCCTCCGAATCGTAACGGA

Non_Targeting_Human

572
1|sg_Non_Targeting_Human_GA_0139|
Non_Targeting_Human
AAACGGTACGACAGCGTGTG

Non_Targeting_Human

573
1|sg_Non_Targeting_Human_GA_0140|
Non_Targeting_Human
ACATAGTCGACGGCTCGATT

Non_Targeting_Human

574
1|sg_Non_Targeting_Human_GA_0141|
Non_Targeting_Human
GATGGCGCTTCAGTCGTCGG

Non_Targeting_Human

575
1|sg_Non_Targeting_Human_GA_0142|
Non_Targeting_Human
ATAATCCGGAAACGCTCGAC

Non_Targeting_Human

576
1|sg_Non_Targeting_Human_GA_0143|
Non Targeting_Human
CGCCGGGCTGACAATTAACG

Non_Targeting_Human

577
1|sg_Non_Targeting_Human_GA_0144|
Non_Targeting_Human
CGTCGCCATATGCCGGTGGC

Non_Targeting_Human

578
1|sg_Non_Targeting_Human_GA_0145|
Non_Targeting_Human
CGGGCCTATAACACCATCGA

Non_Targeting_Human

579
1|sg_Non_Targeting_Human_GA_0146|
Non_Targeting_Human
CGCCGTTCCGAGATACTTGA

Non_Targeting_Human

580
1|sg_Non_Targeting_Human_GA_0147|
Non_Targeting_Human
CGGGACGTCGCGAAAATGTA

Non_Targeting_Human

581
1|sg_Non_Targeting_Human_GA_0148|
Non_Targeting_Human
TCGGCATACGGGACACACGC

Non_Targeting_Human

582
1|sg Non_Targeting_Human_GA_0149|
Non_Targeting_Human
AGCTCCATCGCCGCGATAAT

Non_Targeting_Human

583
1|sg_Non_Targeting_Human_GA_0150|
Non_Targeting_Human
ATCGTATCATCAGCTAGCGC

Non_Targeting_Human

584
1|sg_Non_Targeting_Human_GA_0151|
Non_Targeting_Human
TCGATCGAGGTTGCATTCGG

Non_Targeting_Human

585
1|sg_Non_Targeting_Human_GA_0152|
Non_Targeting Human
CTCGACAGTTCGTCCCGAGC

Non_Targeting_Human

586
1|sg_Non_Targeting_Human_GA_0153|
Non_Targeting_Human
CGGTAGTATTAATCGCTGAC

Non_Targeting_Human

587
1|sg_Non_Targeting_Human_GA_0154|
Non_Targeting_Human
TGAACGCGTGTTTCCTTGCA

Non_Targeting_Human

588
1|sg_Non_Targeting_Human_GA_0155|
Non_Targeting_Human
CGACGCTAGGTAACGTAGAG

Non_Targeting_Human

589
1|sg_Non_Targeting_Human_GA_0156|
Non_Targeting_Human
CATTGTTGAGCGGGCGCGCT

Non_Targeting_Human

590
1|sg_Non_Targeting_Human_GA_0157|
Non_Targeting_Human
CCGCTATTGAAACCGCCCAC

Non_Targeting_Human

591
1|sg_Non_Targeting_Human_GA_0158|
Non_Targeting_Human
AGACACGTCACCGGTCAAAA

Non_Targeting_Human

592
1|sg_Non_Targeting_Human_GA_0159|
Non Targeting Human
TTTACGATCTAGCGGCGTAG

Non_Targeting_Human

593
1|sg_Non_Targeting_Human_GA_0160|
Non_Targeting_Human
TTCGCACGATTGCACCTTGG

Non_Targeting_Human

594
1|sg_Non_Targeting_Human_GA_0161|
Non_Targeting_Human
GGTTAGAGACTAGGCGCGCG

Non_Targeting_Human

595
1|sg_Non_Targeting_Human_GA_0162|
Non_Targeting_Human
CCTCCGTGCTAACGCGGACG

Non_Targeting_Human

596
1|sg_Non_Targeting_Human_GA_0163|
Non_Targeting_Human
TTATCGCGTAGTGCTGACGT

Non_Targeting_Human

597
1|sg_Non_Targeting_Human_GA_0164|
Non_Targeting_Human
TACGCTTGCGTTTAGCGTCC

Non_Targeting_Human

598
1|sg_Non_Targeting_Human_GA_0165|
Non_Targeting_Human
CGCGGCCCACGCGTCATCGC

Non_Targeting_Human

599
1|sg_Non_Targeting_Human_GA_0166|
Non_Targeting_Human
AGCTCGCCATGTCGGTTCTC

Non_Targeting_Human

600
1|sg_Non_Targeting_Human_GA_0167|
Non_Targeting_Human
AACTAGCCCGAGCAGCTTCG

Non_Targeting_Human

601
1|sg_Non_Targeting_Human_GA_0168|
Non_Targeting_Human
CGCAAGGTGTCGGTAACCCT

Non_Targeting_Human

602
1|sg_Non_Targeting_Human_GA_0169|
Non_Targeting_Human
CTTCGACGCCATCGTGCTCA

Non_Targeting_Human

603
1|sg_Non_Targeting_Human_GA_0170|
Non_Targeting_Human
TCCTGGATACCGCGTGGTTA

Non_Targeting_Human

604
1|sg_Non_Targeting_Human_GA_0171|
Non_Targeting_Human
ATAGCCGCCGCTCATTACTT

Non_Targeting_Human

605
1|sg_Non_Targeting_Human_GA_0172|
Non_Targeting_Human
GTCGTCCGGGATTACAAAAT

Non_Targeting_Human

606
1|sg Non_Targeting_Human_GA_0173|
Non_Targeting_Human
TAATGCTGCACACGCCGAAT

Non_Targeting_Human

607
1|sg_Non_Targeting_Human_GA_0174|
Non_Targeting_Human
TATCGCTTCCGATTAGTCCG

Non_Targeting_Human

608
1|sg_Non_Targeting_Human_GA_0175|
Non_Targeting_Human
GTACCATACCGCGTACCCTT

Non_Targeting_Human

609
1|sg_Non_Targeting_Human_GA_0176|
Non_Targeting_Human
TAAGATCCGCGGGTGGCAAC

Non_Targeting_Human

610
1|sg_Non_Targeting_Human_GA_0177|
Non_Targeting_Human
GTAGACGTCGTGAGCTTCAC

Non_Targeting_Human

611
1|sg_Non_Targeting_Human_GA_0178|
Non_Targeting_Human
TCGCGGACATAGGGCTCTAA

Non_Targeting_Human

612
1|sg_Non_Targeting_Human_GA_0179|
Non_Targeting_Human
AGCGCAGATAGCGCGTATCA

Non_Targeting_Human

613
1|sg_Non_Targeting_Human_GA_0180|
Non_Targeting_Human
GTTCGCTTCGTAACGAGGAA

Non_Targeting_Human

614
1|sg_Non_Targeting_Human_GA_0181|
Non_Targeting_Human
GACCCCCGATAACTTTTGAC

Non_Targeting_Human

615
1|sg_Non_Targeting_Human_GA_0182|
Non_Targeting_Human
ACGTCCATACTGTCGGCTAC

Non_Targeting_Human

616
1|sg_Non_Targeting_Human_GA_0183|
Non_Targeting_Human
GTACCATTGCCGGCTCCCTA

Non_Targeting_Human

617
1|sg_Non_Targeting_Human_GA_0184|
Non_Targeting_Human
TGGTTCCGTAGGTCGGTATA

Non_Targeting_Human

618
1|sg_Non_Targeting_Human_GA_0185|
Non_Targeting_Human
TCTGGCTTGACACGACCGTT

Non_Targeting_Human

619
1|sg_Non_Targeting_Human_GA_0186|
Non_Targeting_Human
CGCTAGGTCCGGTAAGTGCG

Non_Targeting_Human

620
1|sg_Non_Targeting_Human_GA_0187|
Non_Targeting_Human
AGCACGTAATGTCCGTGGAT

Non_Targeting_Human

621
1|sg_Non_Targeting_Human_GA_0188|
Non_Targeting_Human
AAGGCGCGCGAATGTGGCAG

Non_Targeting_Human

622
1|sg_Non_Targeting_Human_GA_0189|
Non_Targeting_Human
ACTGCGGAGCGCCCAATATC

Non_Targeting_Human

623
1|sg_Non_Targeting_Human_GA_0190|
Non_Targeting_Human
CGTCGAGTGCTCGAACTCCA

Non_Targeting_Human

624
1|sg_Non_Targeting_Human_GA_0191|
Non_Targeting_Human
TCGCAGCGGCGTGGGATCGG

Non_Targeting_Human

625
1|sg_Non_Targeting_Human_GA_0192|
Non_Targeting_Human
ATCTGTCCTAATTCGGATCG

Non_Targeting_Human

626
1|sg_Non_Targeting_Human_GA_0193|
Non_Targeting_Human
TGCGGCGTAATGCTTGAAAG

Non_Targeting_Human

627
1|sg_Non_Targeting_Human_GA_0194|
Non_Targeting_Human
CGAACTTAATCCCGTGGCAA

Non_Targeting_Human

628
1|sg_Non_Targeting_Human_GA_0195|
Non_Targeting Human
GCCGTGTTGCTGGATACGCC

Non_Targeting_Human

629
1|sg_Non_Targeting_Human_GA_0196|
Non_Targeting_Human
TACCCTCCGGATACGGACTG

Non_Targeting_Human

630
1|sg_Non_Targeting_Human_GA_0197|
Non_Targeting_Human
CCGTTGGACTATGGCGGGTC

Non_Targeting_Human

631
1|sg_Non_Targeting_Human_GA_0198|
Non_Targeting_Human
GTACGGGGCGATCATCCACA

Non_Targeting_Human

632
1|sg_Non_Targeting_Human_GA_0199|
Non_Targeting_Human
AAGAGTAGTAGACGCCCGGG

Non_Targeting_Human

633
1|sg_Non_Targeting_Human_GA_0200|
Non_Targeting_Human
AAGAGCGAATCGATTTCGTG

Non_Targeting_Human

634
3|sg_hCDC16_CC_1|CDC16
CDC16
TCAACACCAGTGCCTGACGG

635
3|sg_hCDC16_CC_2|CDC16
CDC16
AAAGTAGCTTCACTCTCTCG

636
3|sg_hCDC16_CC_3|CDC16
CDC16
GAGCCAACCAATAGATGTCC

637
3|sg_hCDC16_CC_4|CDC16
CDC16
GCGCCGCCATGAACCTAGAG

638
3|sg_hGTF2B_CC_1|GTF2B
GTF2B
ACAAAGGTTGGAACAGAACC

639
3|sg_hGTF2B_CC 2|GTF2B
GTF2B
GGTGACCGGGTTATTGATGT

640
3|sg_hGTF2B_CC_3|GTF2B
GTF2B
TTAGTGGAGGACTACAGAGC

641
3|sg_hGTF2B_CC_4|GTF2B
GTF2B
ACATATAGCCCGTAAAGCTG

642
3|sg_hHSPA5_CC_1|HSPA5
HSPA5
CGTTGGCGATGATCTCCACG

643
3|sg_hHSPA5_CC_2|HSPA5
HSPA5
TGGCCTTTTCTACCTCGCGC

644
3|sg_hHSPA5_CC_3|HSPA5
HSPA5
AATGGAGATACTCATCTGGG

645
3|sg_hHSPA5_CC_4|HSPA5
HSPA5
GAAGCCCGTCCAGAAAGTGT

646
3|sg_hHSPA9_CC_1|HSPA9
HSPA9
CAATCTGAGGAACTCCACGA

647
3|sg_hHSPA9_CC_2|HSPA9
HSPA9
AGGCTGCGGCGCCCACGAGA

648
3|sg_hHSPA9_CC_3|HSPA9
HSPA9
ACTTTGACCAGGCCTTGCTA

649
3|sg_hHSPA9_CC_4|HSPA9
HSPA9
ACCTTCCATAACTGCCACGC

650
3|sg_hPAFAH1B1_CC_1|PAFAH1B1
PAFAH1B1
CGAGGCGTACATACCCAAGG

651
3|sg hPAFAH1B1_CC_2|PAFAH1B1
PAFAH1B1
ATGGTACGGCCAAATCAAGA

652
3|sg hPAFAH1B1_CC_3|PAFAH1B1
PAFAH1B1
TCTTGTAATCCCATACGCGT

653
3|sg_hPAFAH1B1_CC_4|PAFAH1B1
PAFAH1B1
ATTCACAGGACACAGAGAAT

654
3|sg_hPCNA_CC_1|PCNA
PCNA
CCAGGGCTCCATCCTCAAGA

655
3|sg_hPCNA_CC_2|PCNA
PCNA
TGAGCTGCACCAAAGAGACG

656
3|sg_hPCNA_CC_3|PCNA
PCNA
ATGTCTGCAGATGTACCCCT

657
3|sg_hPCNA_CC_4|PCNA
PCNA
CGAAGATAACGCGGATACCT

658
3|sg_hPOLR2L_CC_1|POLR2L
POLR2L
GCTGCAGGCCGAGTACACCG

659
3|sg_hPOLR2L_CC_2|POLR2L
POLR2L
ACAAGTGGGAGGCTTACCTG

660
3|sg_hPOLR2L_CC_3|POLR2L
POLR2L
GCAGCGTACAGGGATGATCA

661
3|sg_hPOLR2L_CC_4|POLR2L
POLR2L
GCAGTAGCGCTTCAGGCCCA

662
3|sg_hRPL9_CC_1|RPL9
RPL9
CAAATGGTGGGGTAACAGAA

663
3|sg_hRPL9_CC_2|RPL9
RPL9
GAAAGGAACTGGCTACCGTT

664
3|sg_hRPL9_CC_3|RPL9
RPL9
AGGGCTTCCGTTACAAGATG

665
3|sg_hRPL9_CC_4|RPL9
RPL9
GAACAAGCAACACCTAAAAG

666
3|sg_hSF3A3_CC_1|SF3A3
SF3A3
TGAGGAGAAGGAACGGCTCA

667
3|sg_hSF3A3_CC_2|SF3A3
SF3A3
GGAAGAATGCAGAGTATAAG

668
3|sg_hSF3A3_CC_3|SF3A3
SF3A3
GGAATTTGAGGAACTCCTGA

669
3|sg_hSF3A3_CC_4|SF3A3
SF3A3
GCTCACCGGCCATCCAGGAA

670
3|sg_hSF3B3_CC_1|SF3B3
SF3B3
ACTGGCCAGGAACGATGCGA

671
3|sg_hSF3B3_CC_2|SF3B3
SF3B3
GCAGCTCCAAGATCTTCCCA

672
3|sg_hSF3B3_CC_3|SF3B3
SF3B3
GAATGAGTACACAGAACGGA

673
3|sg_hSF3B3_CC_4|SF3B3
SF3B3
GGAGCAGGACAAGGTCGGGG

Example 2—BRD9 Degrader Depletes BRD9 Protein

The following example demonstrates the depletion of the BRD9 protein in synovial sarcoma cells treated with a BRD9 degrader.

Procedure: Cells were treated with DMSO or the BRD9 degrader, Compound 1 (also known as dBRD9, see Remillard et al, Angew. Chem. Int. Ed. Engl. 56(21):5738-5743 (2017); see structure of compound 1 below), for indicated doses and timepoints.

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Whole cell extracts were fractionated by SDS-PAGE and transferred to a polyvinylidene difluoride membrane using a transfer apparatus according to the manufacturer's protocols (Bio-Rad). After incubation with 5% nonfat milk in TBST (10 mM Tris, pH 8.0, 150 mM NaCl, 0.5% Tween 20) for 60 min, the membrane was incubated with antibodies against BRD9 (1:1,000, Bethyl laboratory A303-781A), GAPDH (1:5,000, Cell Signaling Technology), and/or MBP (1:1,000, BioRad) overnight at 4° C. Membranes were washed three times for 10 minutes and incubated with anti-mouse or anti-rabbit antibodies conjugated with either horseradish peroxidase (HRP, FIGS. 2-3) or IRDye (FIG. 4, 1:20,000, LI-COR) for at least 1 h. Blots were washed with TBST three times and developed with either the ECL system according to the manufacturer's protocols (FIGS. 2-3) or scanned on an Odyssey CLx Imaging system (FIG. 4).

Results: Treatment of SYO1 synovial sarcoma cells with the BRD9 degrader Compound 1 results in dose dependent (FIG. 2) and time dependent (FIG. 3) depletion of BRD9 in the cells. Further, as shown in FIG. 4, the depletion of BRD9 by Compound 1 is replicated in a non-synovial sarcoma cell line (293T) and may be sustained for at least 5 days.

Example 3—Inhibition of Growth of Synovial Cell Lines by BRD9 Inhibitors and BRD9 Degraders

The following example demonstrates that BRD9 degraders and inhibitors selectively inhibit growth of synovial sarcoma cells.

Procedures:

Cells were treated with DMSO or the BRD9 degrader, Compound 1, at indicated concentrations, and proliferation was monitored from day 7 to day 14 by measuring confluency over time using an IncuCyte live cell analysis system (FIG. 5). Growth medium and compounds were refreshed every 3-4 days.

Cells were seeded into 12-well plates and treated with DMSO, 1 μM BRD9 inhibitor, Compound 2 (also known as BI-7273, see Martin et al, J Med Chem. 59(10):4462-4475 (2016); see structure of compound 2 below), or 1 μM BRD9 degrader, Compound 1.

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The number of cells was optimized for each cell line. Growth medium and compounds were refreshed every 3-5 days. SYO1, Yamato, A549, 293T and HS-SY-II cells were fixed and stained at day 11. ASKA cells were fixed and stained at day 23. Staining was done by incubation with crystal violet solution (0.5 g Crystal Violet, 27 ml 37% Formaldehyde, 100 mL 10×PBS, 10 mL Methanol, 863 dH2O to 1 L) for 30 minutes followed by 3× washes with water and drying the plates for at least 24h at room temperature. Subsequently plates were scanned on an Odyssey CLx Imaging system (FIG. 6).

Cells were seeded into 96-well ultra low cluster plate (Costar, #7007) in 200 μL complete media and treated at day 2 with DMSO, Staurosporin, or BRD9 degrader, Compound 1, at indicated doses (FIG. 7). Media and compounds were changed every 5 d and cell colonies were imaged at day 14.

Results: As shown in FIGS. 5, 6, and 7, treatment of synovial sarcoma cell lines (SYO1, Yamato, HS-SY-II, and ASKA) with a BRD9 inhibitor, Compound 2, or a BRD9 degrader, Compound 1, results in inhibition of the growth of the cells, but does not result in inhibition of the growth of non-synovial control cancer cell lines (293T, A549, G401).

Example 4—Selective Inhibition of Growth of Synovial Cell Lines by BRD9 Degraders and BRD9 Binders

The following example demonstrates that BRD9 degraders and binders selectively inhibit growth of synovial sarcoma cells.

Procedure: Cells were seeded into 6-well or 12-well plates and were treated daily with a BRD9 degrader (Compound 1), a bromo-domain BRD9 binder (Compound 2), E3 ligase binder (lenalidomide), DMSO, or staurosporin (positive control for cell killing), at indicated concentrations. The number of cells was optimized for each cell line. Growth media was refreshed every 5 days. By day 14, medium was removed, cells were washed with PBS, and stained using 500 μL of 0.005% (w/v) crystal violet solution in 25% (v/v) methanol for at least 1 hour at room temperature. Subsequently plates were scanned on an Odyssey CLx Imaging system.

Results: As shown in FIGS. 8 and 9, treatment of synovial sarcoma cell lines (SYO1, HS-SY-II, and ASKA) with Compound 1 or Compound 2 resulted in inhibition of the growth of the cells, but did not result in inhibition of the growth of non-synovial control cancer cell lines (RD, HCT116, and Calu6). Overall, Compound 1 showed most significant growth inhibition in all synovial cell lines.

Example 5—Inhibition of Cell Growth in Synovial Sarcoma Cells

The following example shows that BRD9 degraders inhibit cell growth and induce apoptosis in synovial sarcoma cells.

Procedure: SYO1 cells were treated for 8 or 13 days with DMSO, a BRD9 degrader (Compound 1) at 200 nM or 1 μM, or an E3 ligase binder (lenalidomide) at 200 nM. Compounds were refreshed every 5 days. Cell cycle analysis was performed using the Click-iT™ Plus EdU Flow Cytometry Assay (Invitrogen). The apoptosis assay was performed using the Annexin V-FITC Apoptosis Detection Kit (Sigma A9210). Assays were performed according to the manufacturer's protocol.

Results: As shown in FIGS. 10-13, treatment with Compound 1 for 8 or 13 days resulted in reduced numbers of cells in the S-phase of the cell cycle as compared to DMSO and lenalidomide. Treatment with Compound 1 for 8 days also resulted in increased numbers of early- and late-apoptotic cells as compared to DMSO controls.

Example 6—Composition for SS18-SSX1-BAF

The following example shows the identification of BRD9 as a component of SS18-SSX containing BAF complexes.

Procedure: A stable 293T cell line expressing HA-SS18SSX1 was generated using lentiviral integration. SS18-SSX1 containing BAF complexes were subject to affinity purification and subsequent mass spectrometry analysis revealed SS18-SSX1 interacting proteins.

Results: As shown in FIG. 14, BAF complexes including the SS18-SSX fusion protein also included BRD9. More than 5 unique peptides were identified for ARID1A (95 peptides), ARID1B (77 peptides), SMARCC1 (69 peptides), SMARCD1 (41 peptides), SMARCD2 (37 peptides), DPF2 (32 peptides), SMARCD3 (26 peptides), ACTL6A (25 peptides), BRD9 (22 peptides), DPF1 Isoform 2 (18 peptides), DPF3 (13 peptides), and ACTL6B (6 peptides).

Example 7—Preparation of N-(1,1-dioxo-1λ6,2-thiazinan-4-yl)-5-methyl-4-oxo-7-[3-(trifluoromethyl)phenyl]thieno[3,2-c]pyridine-2-carboximidamide (Compound B1)

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Step 1: Preparation of 4-amino-1λ6,2-thiazinane-1,1-dione (i-1)

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4-(benzylamino)-1λ6,2-thiazinane-1,1-dione (200.00 mg, 0.832 mmol, 1.00 equiv) and Pd/C (199.27 mg, 1.873 mmol, 2.25 equiv) in MeOH (10.00 mL) were stirred under an atmosphere of hydrogen at balloon and room temperature for 3 hours. The solid was filtered out, and the filtrate was concentrated under reduced pressure to afford 4-amino-1λ6,2-thiazinane-1,1-dione (120 mg, 96%) as a brown solid. This material was used directly in the next step without further purification. LCMS (ESI) m/z: [M+H]+=151.

Step 2: Preparation of 4-amino-1λ6,2-thiazinane-1,1-dione Hydrochloride (i-2)

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4-amino-1λ6,2-thiazinane-1,1-dione (110.00 mg, 0.732 mmol, 1.00 equiv) was added a solution of HCl in 1,4-dioxane (4 mL, 4 M), and the resulting mixture was stirred at room temperature for 2 hours. The solvent was removed under reduced pressure to afford 4-amino-1λ6,2-thiazinane-1,1-dione hydrochloride (131 mg, 96%) as a brown solid. This material was used directly in the next step without further purification. LCMS (ESI) m/z: [M+H]+=151.

Step 3: Preparation of N-(1,1-dioxo-1λ6,2-thiazinan-4-yl)-5-methyl-4-oxo-7-[3-(trifluoromethyl)phen yl]thieno[3,2-c]pyridine-2-carboximidamide (Compound B1)

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To a solution of 5-methyl-4-oxo-7-[3-(trifluoromethyl)phenyl]thieno[3,2-c]pyridine-2-carbonitrile (50.00 mg, 0.150 mmol, 1.00 equiv) in MeOH (1.00 mL) was added NaOMe (8.08 mg, 0.150 mmol, 1 equiv), and the resulting mixture was stirred at 75° C. for 3 hours. Then 4-amino-1λ6,2-thiazinane-1,1-dione hydrochloride (33.50 mg, 0.179 mmol, 1.2 equiv) was added to the reaction mixture. After stirring at 75° C. for 16 hours, the crude product was purified by flash C18-flash chromatography, elution gradient 0 to 50% MeCN in water (containing 0.1% NH₄HCO₃). Pure fractions were evaporated to dryness to afford crude product. The crude product was purified by preparative HPLC (conditions: XBridge Prep OBD 018 Column, 19*250 mm, 5 μm; Mobile Phase A: Water (10 mmol/L NH₄HCO₃), Mobile Phase B: ACN; Flow rate: 25 mL/minute; Gradient: 35 B to 50 B in 12 minutes; 254/220 nm; R_T(retention time): 8.62 minutes). Fractions containing the desired compound were evaporated to dryness to afford N-(1,1-dioxo-1λ6,2-thiazinan-4-yl)-5-methyl-4-oxo-7-[3-(trifluoromethyl)phenyl]thieno[3,2-c]pyridine-2-carboximidamide (5.8 mg, 7.9%) as a white solid. LCMS (ESI) m/z: [M+H]+=485.20.

Example 8—Preparation of 7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-2-[3H-imidazo[4,5-c]pyridin-2-yl]-5-methylthieno[3,2-c]pyridin-4-one (Compound B2)

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Step 1: Preparation of 5-methyl-4-oxothieno[3,2-c]pyridine-2-carbaldehyde (i-6)

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To a solution of 2-bromo-5-methylthieno[3,2-c]pyridin-4-one (2.00 g, 8.193 mmol, 1.00 equiv) in THF (20.00 mL) was added n-BuLi (1.05 g, 16.386 mmol, 2.00 equiv) in dropwise at −78° C. under nitrogen atmosphere, the resulting mixture was stirred at −78° C. for 30 minutes, then DMF (5.99 g, 81.930 mmol, 10.00 equiv) was added to the reaction mixture. The reaction was warmed to room temperature naturally. The reaction was quenched by the addition of sat. NH₄Cl (aq.) (30 mL) at 0° C., and extracted with EtOAc (ethyl acetate) (50 mL×4). The organic layer was washed with water (50 mL) and brine (50 mL), then dried over anhydrous sodium sulfate, filtered, and concentrated to give crude product that was purified by flash silica chromatography, elution gradient 0 to 60% EtOAc in petroleum ether (PE). Pure fractions were evaporated to dryness to afford 5-methyl-4-oxothieno[3,2-c]pyridine-2-carbaldehyde (658 mg, 42%) as a brown solid. LCMS (ESI) m/z: [M+H]⁺=194.

Step 2: Preparation of 7-bromo-5-methyl-4-oxothieno[3,2-c]pyridine-2-carbaldehyde (i-7)

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To a solution of 5-methyl-4-oxothieno[3,2-c]pyridine-2-carbaldehyde (600.00 mg, 3.105 mmol, 1.00 equiv) in THF (10.00 mL) was added NBS (663.23 mg, 3.726 mmol, 1.20 equiv), the resulting mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with EtOAc (50 mL), and washed with water (3×30 mL) and saturated brine (1×30 mL). The organic layer was dried over Na₂SO₄, filtered and evaporated to afford crude product. The crude product was purified by flash silica chromatography, elution gradient 0 to 60% EtOAc in petroleum ether. Pure fractions were evaporated to dryness to afford 7-bromo-5-methyl-4-oxothieno[3,2-c]pyridine-2-carbaldehyde (510 mg, 60%) as a brown solid. LCMS (ESI) m/z: [M+H]⁺=272.

Step 3: Preparation of 7-bromo-2-[3H-imidazo[4,5-c]pyridin-2-yl]-5-methylthieno[3,2-c]pyridin-4-one (i-9)

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To a solution of 7-bromo-5-methyl-4-oxothieno[3,2-c]pyridine-2-carbaldehyde (300.00 mg, 1.102 mmol, 1.00 equiv) and 3,4-diaminopyridine (120.31 mg, 1.102 mmol, 1 equiv) in DMF (5.00 mL) was added Na₂S₂O₅(628.75 mg, 3.307 mmol, 3 equiv), the resulting mixture was stirred at 120° C. for 16 hours. The solvent was removed under reduced pressure. The residue was purified by flash silica chromatography, elution gradient 0 to 10% MeOH in DCM. Pure fractions were evaporated to dryness to afford 7-bromo-2-[3H-imidazo[4,5-c]pyridin-2-yl]-5-methylthieno[3,2-c]pyridin-4-one (165 mg, 41%) as a brown solid. LCMS (ESI) m/z: [M+H]⁺=361.

Step 4: Preparation of 7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-2-[3H-imidazo[4,5-c]pyridin-2-yl]-5-methylthieno[3,2-c]pyridin-4-one (Compound 82)

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To a solution of 7-bromo-2-[3H-imidazo[4,5-c]pyridin-2-yl]-5-methylthieno[3,2-c]pyridin-4-one (60.00 mg, 0.166 mmol, 1.00 equiv), 4-[(dimethylamino)methyl]-3,5-dimethoxyphenylboronic acid (47.65 mg, 0.199 mmol, 1.20 equiv) and 052003 (108.24 mg, 0.332 mmol, 2.00 equiv) in DMF (1.00 mL) and H₂O (0.25 mL) was added Pd(dppf)Cl₂(12.15 mg, 0.017 mmol, 0.10 equiv), the resulting mixture was stirred at 100° C. for 1 hour under a nitrogen atmosphere. The mixture was concentrated, the crude product was purified by reverse phase flash with the following conditions to afford crude product: column, 018 silica gel; mobile phase, MeCN in water, 5% to 50% gradient in 10 min; detector, UV 254 nm. The crude product was purified by Prep-HPLC (conditions: XSelect CSH Prep C18 OBD Column, 5 μm, 19*150 mm; Mobile Phase A: Water (0.05% TFA, trifluoroacetic acid); Mobile Phase B: ACN (acetonitrile); Flow rate: 25 mL/minute; Gradient: 8% B to 8% B in 2 minutes; 254/220 nm; R_T(retention time): 13.87 minutes) to afford 7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-2-[3H-imidazo[4,5-c]pyridin-2-yl]-5-methylthieno[3,2-c]pyridin-4-one (3 mg, 3.8%) as a yellow solid. ¹H NMR (400 MHz, Methanol-d4) δ 9.19 (s, 1H), 8.55 (d, J=6.5 Hz, 1H), 8.46 (s, 1H), 8.10 (d, J=6.5 Hz, 1H), 7.93 (s, 1H), 7.11 (s, 2H), 4.44 (s, 2H), 4.05 (s, 6H), 3.75 (s, 3H), 2.94 (s, 6H). LCMS (ESI) m/z: [M+H]⁺=476.30.

Example 9—Preparation of N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-2-yl)-1-methylpiperidine-4-carboxamide (Compound B3)

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Step 1: Preparation of 2-bromo-7-chloro-5-methylthieno[3,2-c]pyridin-4-one (i-11)

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To a solution of 2-bromo-5-methylthieno[3,2-c]pyridin-4-one (1.50 g, 6.145 mmol, 1.00 equiv) in DMF (5.00 mL) was added NCS (0.90 mg, 0.007 mmol, 1.1 equiv) at 25° C. The resulting solution was stirred at 70° C. for 3 hours. The resulting mixture was concentrated. The residue was purified by chromatography on silica gel eluted with petroleum ether/EtOAc (2/1) to afford 2-bromo-7-chloro-5-methylthieno[3,2-c]pyridin-4-one (1.6 g, 94%) as a brown solid. LCMS (ESI) m/z: [M+H]+=278

Step 2: Preparation of 7-chloro-2-[(diphenylmethylidene)amino]-5-methylthieno[3,2-c]pyridin-4-one (i-12)

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To a solution of 2-bromo-7-chloro-5-methylthieno[3,2-c]pyridin-4-one (1.6 g, 5.744 mmol, 1.00 equiv) and benzophenone imine (1093.09 mg, 6.032 mmol, 1.05 equiv) in dioxane (15 mL) was added Xantphos (332.37 mg, 0.574 mmol, 0.1 equiv), sodium tert-butoxide (1656.09 mg, 17.232 mmol, 3 equiv), and Pd₂(dba)₃(262.99 mg, 0.288 mmol, 0.05 equiv) at 25° C. The resulting solution was stirred for 2 hours at 80° C. The resulting mixture was concentrated. The residue was purified by chromatography on silica gel eluted with petroleum ether/EtOAc (5:1) to afford 7-chloro-2-[(diphenylmethylidene)amino]-5-methylthieno[3,2-c]pyridin-4-one (1.6 g, 74%) as a dark yellow solid. LCMS (ESI) m/z: [M+H]+=379.

Step 3: Preparation of 2-amino-7-chloro-5-methylthieno[3,2-c]pyridin-4-one (i-13)

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To a solution of 7-chloro-2-[(diphenylmethylidene)amino]-5-methylthieno[3,2-c]pyridin-4-one (1.60 g, 4.223 mmol, 1.00 equiv) in THF (10.00 mL) was added HCl (5.00 mL) at 25° C. The resulting solution was stirred at 25° C. for 2 hours. The solvent was removed under reduced pressure. This resulted in 1.5 g (crude) of 2-amino-7-chloro-5-methylthieno [3,2-c]pyridin-4-one as a dark brown solid. The product was used in the next step directly without further purification. LCMS (ESI) m/z: [M+H]+=215.

Step 4: Preparation of N-[7-chloro-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl]-1-methylpiperidine-4-carboxamide (i-14)

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To a solution of 1-methylpiperidine-4-carboxylic acid (80.04 mg, 0.559 mmol, 1.20 equiv) and HATU (354.25 mg, 0.932 mmol, 2.00 equiv) in DMF (1.00 mL) was added 2-amino-7-chloro-5-methylthieno[3,2-c]pyridin-4-one (100.00 mg, 0.466 mmol, 1.00 equiv) and DIEA (180.62 mg, 1.397 mmol, 3.00 equiv), the resulting solution was stirred at 25 degree for 2 hours. The resulting mixture was concentrated. The residue was applied onto a silica gel column with CH₂Cl₂/MeOH (20:1). This resulted in (80 mg, 51%) of N-[7-chloro-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl]-1-methylpiperidine-4-carboxamide as a yellow solid. LCMS (ESI) m/z: [M+H]+=340.10

Step 5: Preparation of N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-2-yl)-1-methylpiperidine-4-carboxamide (Compound 83

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To a solution of N-[7-chloro-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-2-yl]-1-methylpiperidine-4-carboxamide (60.00 mg, 0.177 mmol, 1.00 equiv) and [4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]boronic acid (50.65 mg, 0.212 mmol, 1.20 equiv) in H₂O (1.00 mL) and dioxane (3.00 mL) was added 052003 (172.57 mg, 0.530 mmol, 3.00 equiv) and Pd(dppf)Cl₂.CH₂Cl₂(12.92 mg, 0.018 mmol, 0.10 equiv). The resulting solution was stirred at 80° C. for 2 hours (under N₂atmosphere). The crude product was purified by preparative HPLC (conditions: XSelect CSH Prep C18 OBD Column, 19*150 mm; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 25 mL/minute; Gradient: 20% B to 55% B in 8 minutes; 254 nm; R_T: 7.12 minutes). Fractions containing the desired compound were evaporated to dryness to afford N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-2-yl)-1-methylpiperidine-4-carboxamide (32 mg, 36%) as a white solid. ¹H NMR (400 MHz, Methanol-d4) δ 7.68 (s, 1H), 7.23 (s, 1H), 7.06 (s, 2H), 4.42 (s, 2H), 4.02 (s, 6H), 3.75 (s, 3H), 3.64 (d, J=12.6 Hz, 2H), 3.16-3.06 (m, 2H), 2.92 (d, J=1.0 Hz, 9H), 2.82-2.71 (m, 1H), 2.21 (d, J=14.5 Hz, 2H), 2.10-1.94 (m, 2H). LCMS (ESI) m/z: [M+H]+=499.30.

Example 10—Preparation of N-(7-[4-[(Dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-2-yl)-4-methylmorpholine-2-carboxamide Formic Acid (Compound B4 Formic Acid)

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Step 1: Preparation of tert-butyl 2-([7-chloro-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl]carbamoyl l)morpholine-4-carboxylate (i-15)

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To a solution of 2-amino-7-chloro-5-methylthieno[3,2-c]pyridin-4-one (507.00 mg, 2.362 mmol, 1.00 equiv) and 4-(tert-butoxycarbonyl)morpholine-2-carboxylic acid (710.00 mg, 3.070 mmol, 1.3 equiv)) in DMF (6 mL) was added HATU (1796.03 mg, 4.724 mmol, 2 equiv) and DIEA (1220.97 mg, 9.447 mmol, 4 equiv) at 25° C. The resulting solution was stirred at 25° C. for 2 hours. The resulting mixture was concentrated. The residue was purified by chromatography on silica gel eluted with petroleum ether/EtOAc (2:1) to afford tert-butyl2-([7-chloro-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl]carbamoyl)morpholine-4-carboxylate (820 mg, 81%) as a brown solid. LCMS (ESI) m/z: [M+H]+=428.

Step 2: Preparation of N-[7-chloro-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl]morpholine-2-carboxamide (i-16)

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To a solution of tert-butyl 2-([7-chloro-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl]carbamoyl)morpholine-4-carboxylate (800.00 mg, 1.870 mmol, 1.00 equiv) in DCM (6.00 mL) was added TFA (2.00 mL) at 25° C. The resulting solution was stirred at 25° C. for 2 hours. The solvent was removed under reduced pressure. This resulted in 1 g (crude) of N-[7-chloro-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl]morpholine-2-carboxamide as a dark brown solid. The product was used in the next step directly without further purification. LCMS (ESI) m/z: [M+H]+=328.

Step 3: Preparation of N-[7-chloro-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl]-4-methylmorpholine-2-carboxamide (i-17)

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To a solution of N-[7-chloro-5-methyl-4-oxothie[3,2-c]pyridin-2-yl]morpholine-2-carboxamide (1.00 g, 3.051 mmol, 1.00 equiv) in HCHO (4.00 mL) and MeOH (10.00 mL) was added NaBH₃CN (0.38 g, 0.006 mmol, 2 equiv) and CH₃COOH (0.50 mL, 8.726 mmol, 2.86 equiv) at 25° C. The resulting solution was stirred at 25° C. for 4 hours. The resulting mixture was concentrated. The residue was purified by chromatography on silica gel eluted with petroleum ether/EtOAc (1:1) to afford N-[7-chloro-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl]-4-methylmorpholine-2-carboxamide (284 mg, 27%) as a brown solid. LCMS (ESI) m/z: [M+H]+=342.

Step 4: Preparation of N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-2-yl)-4-methylmorpholine-2-carboxamide Formic Acid (Compound 84 Formic Acid)

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To a solution of N-[7-chloro-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-2-yl]-4-methylmorpholine-2-carboxamide (100 mg, 0.293 mmol, 1.00 equiv) and [4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]boronic acid (104.92 mg, 0.439 mmol, 1.5 equiv) in H₂O (1 mL) and dioxane (4 mL) was added 052003 (190.64 mg, 0.585 mmol, 2 equiv) and XPhos palladium(II) biphenyl-2-amine chloride (23.02 mg, 0.029 mmol, 0.10 equiv) at 25° C. The resulting solution was stirred at 90° C. for 12 hours. The crude product was purified by Prep-HPLC (conditions: Sun Fire Prep 018 OBD Column 19×150 mm 10 nm; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 25 mL/minute; Gradient: 3% B to 3% B in 2 minutes; 254/220 nm; R_T: 7.5 minutes) to afford N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-2-yl)-4-methylmorpholine-2-carboxamide; formic acid (31.7 mg, 20%) as a light brown solid. ¹H NMR (400 MHz, Methanol-d4) δ 8.46 (s, 1H), 7.67 (s, 1H), 7.39 (d, J=0.9 Hz, 1H), 7.06 (d, J=1.1 Hz, 2H), 4.41 (s, 2H), 4.27 (dd, J=10.3, 2.8 Hz, 1H), 4.14-4.01 (m, 1H), 4.02 (s, 6H), 3.79 (td, J=11.4, 2.5 Hz, 1H), 3.74 (s, 3H), 3.12 (d, J=11.7 Hz, 1H), 2.91 (s, 6H), 2.77 (d, J=11.9 Hz, 1H), 2.38 (s, 3H), 2.28 (t, J=11.6 Hz, 1H), 2.17 (t, J=10.9 Hz, 1H). LCMS (ESI) m/z: [M+H]+=501.35.

Example 11—Preparation of 7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxo-N-(6-oxopiperidin-3-yl)-4H,5H-thieno[3,2-c]pyridine-2-carboxamide (Compound B5)

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Step 1: Preparation of Methyl 5-methyl-4-oxo-4,5-dihydrothieno[3,2-c]pyridine-2-carboxylate (i-18)

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To a mixture of 2-bromo-5-methylthieno[3,2-c]pyridin-4-one (2.44 g, 9.996 mmol, 1.00 equiv) and Pd(dppf)Cl₂.CH₂Cl₂(0.82 g, 1.00 mmol, 0.100 equiv) in MeOH (20.00 mL) was added TEA (2.78 mL, 27.460 mmol, 2.00 equiv). The resulting mixture was stirred at 100° C. for 16 hours under CO atmosphere (50 atm). Then the mixture was concentrated under reduced pressure, and the resulting residue was purified by flash silica chromatography, elution gradient 0 to 70% EtOAc in petroleum ether to give methyl 5-methyl-4-oxo-4,5-dihydrothieno[3,2-c]pyridine-2-carboxylate (1.81 g, 81%) as a yellow solid. LCMS (ESI) m/z: [M+H]⁺=224.

Step 2: Preparation of Methyl 7-bromo-5-methyl-4-oxo-4,5-dihydrothieno[3,2-c]pyridine-2-carboxylate (i-19)

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To a stirred mixture of methyl 5-methyl-4-oxothieno[3,2-c]pyridine-2-carboxylate (1.80 g, 8.063 mmol, 1.00 equiv) in) DMF (30 mL) was added NBS (1.58 g, 8.869 mmol, 1.10 equiv). The resulting mixture was stirred at room temperature for 1 hour under nitrogen atmosphere. The mixture was then diluted with water (100 mL) and extracted with EtOAc (300 mL×3). The organic layers were combined and dried over anhydrous Na₂SO₄, filtered, and concentrated to give methyl 7-bromo-5-methyl-4-oxo-4,5-dihydrothieno[3,2-c]pyridine-2-carboxylate (2.43 g, crude) as a yellow solid. This material was used directly in the next step without further purification. LCMS (ESI) m/z: [M+H]⁺=302.

Step 3: Preparation of 7-bromo-5-methyl-4-oxo-4,5-dihydrothieno[3,2-c]pyridine-2-carboxylic Acid (i-20)

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Into a sealed tube was added methyl 7-bromo-5-methyl-4-oxothieno[3,2-c]pyridine-2-carboxylate (2.43 g, 8.05 mmol, 1.00 equiv) and concentrated hydrochloric acid (40 mL). The mixture was stirred for 4 hours at 90° C. The resulting mixture was concentrated to give 7-bromo-5-methyl-4-oxo-4,5-dihydrothieno[3,2-c]pyridine-2-carboxylic acid (2.20 g, 95%) as a brown solid. This material was used directly in the next step without further purification. LCMS (ESI) m/z: [M+H]⁺=288.

Step 4: Preparation of 7-bromo-5-methyl-4-oxo-N-(6-oxopiperidin-3-yl)thieno[3,2-c]pyridine-2-carboxamide (i-21)

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To a solution of 7-bromo-5-methyl-4-oxothieno[3,2-c]pyridine-2-carboxylic acid (50.00 mg, 0.174 mmol, 1.00 equiv) and HATU (131.97 mg, 0.347 mmol, 2.00 equiv) in DMF (1.00 mL) was added 5-aminopiperidin-2-one (21.79 mg, 0.191 mmol, 1.10 equiv) and DIEA (67.29 mg, 0.521 mmol, 3.00 equiv). The resulting solution was stirred at 25° C. for 2 hours. The resulting mixture was concentrated. The residue was applied onto a silica gel column with CH₂Cl₂/MeOH (20:1) to afford 7-bromo-5-methyl-4-oxo-N-(6-oxopiperidin-3-yl)thieno[3,2-c]pyridine-2-carboxamide (42 mg, 63%) as a yellow solid. LCMS (ESI) m/z: [M+H]+=386.01.

Step 5: Preparation of 7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxo-N-(6-oxopiperidin-3-yl)-4H,5H-thieno[3,2-c]pyridine-2-carboxamide (Compound 85)

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To a solution of 7-bromo-5-methyl-4-oxo-N-(6-oxopiperidin-3-yl)-4H,5H-thieno[3,2-c]pyridine-2-carboxamide (30.00 mg, 0.078 mmol, 1.00 equiv) and [4-[(dimethylamino)methyl]-3,5-dimethoxy phenyl]boronic acid (22.40 mg, 0.094 mmol, 1.20 equiv) in H₂O (1.00 mL) and dioxane (3.00 mL) was added Cs₂CO₃(76.31 mg, 0.234 mmol, 3.00 equiv) and Pd(dppf)Cl₂.CH₂Cl₂(6.38 mg, 0.008 mmol, 0.10 equiv). The resulting solution was stirred at 80° C. for 2 hours (under N₂atmosphere). The crude product was purified by preparative HPLC (conditions: XSelect CSH Prep 018 OBD Column, 5 μm, 19150 mm; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 25 mL/minute; Gradient: 20% B to 55% B in 8 minutes; 254 nm; R_T: 7.12 minutes). Fractions containing the desired compound were evaporated to dryness to afford 7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxo-N-(6-oxopiperidin-3-yl)-4H,5H-thieno[3,2-c]pyridine-2-carboxamide (28 mg, 72%) as a white solid. ¹H NMR (400 MHz, Methanol-d4) δ 8.57 (s, 1H, FA), 8.28 (s, 1H), 7.86 (s, 1H), 7.06 (s, 2H), 4.37 (s, 2H), 4.37-4.30 (m, 1H), 4.02 (s, 6H), 3.75 (s, 3H), 3.58 (dd, J=12.0, 4.9 Hz, 1H), 3.30-3.26 (m, 1H), 2.88 (s, 6H), 2.55-2.47 (m, 2H), 2.19-2.01 (m, 2H). LCMS (ESI) m/z: [M+H]+=499.35.

Example 12—Preparation of 7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-N-(1-methylpiperidin-4-yl)-4-oxo-4H,5H-thieno[3,2-c]pyridine-2-carboxamide (Compound B6)

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Step 1: Preparation of 7-bromo-5-methyl-N-(1-methylpiperidin-4-yl)-4-oxothieno[3,2-c]pyridine-2-carboxamide (i-22)

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To a solution of 7-bromo-5-methyl-4-oxothieno[3,2-c]pyridine-2-carboxylic acid (50.00 mg, 0.174 mmol, 1.00 equiv) and HATU (131.97 mg, 0.347 mmol, 2.00 equiv) in DMF (1.00 mL) was added 1-methylpiperidin-4-amine (21.80 mg, 0.191 mmol, 1.10 equiv) and DIEA (67.29 mg, 0.521 mmol, 3.00 equiv). The resulting solution was stirred at 25° C. for 2 hours. The resulting mixture was concentrated. The residue was applied onto a silica gel column with CH₂Cl₂/MeOH (20:1). This resulted in (45 mg, 68%) of 7-bromo-5-methyl-N-(1-methylpiperidin-4-yl)-4-oxothieno[3,2-c]pyridine-2-carboxamide as a yellow solid. LCMS (ESI) m/z: [M+H]+=384.01.

Step 2: Preparation of 7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-N-(1-methylpiperidin-4-yl)-4-oxo-4H,5H-thieno[3,2-c]pyridine-2-carboxamide (Compound 86)

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To a solution of 7-bromo-5-methyl-N-(1-methylpiperidin-4-yl)-4-oxo-4H,5H-thieno[3,2-c]pyridine-2-carboxamide (30.00 mg, 0.078 mmol, 1.00 equiv) and [4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]boronic acid (22.40 mg, 0.094 mmol, 1.20 equiv) in H₂O (1.00 mL) and dioxane (3.00 mL) was added Cs₂CO₃(76.31 mg, 0.234 mmol, 3.00 equiv) and Pd(dppf)Cl₂.CH₂Cl₂(6.38 mg, 0.008 mmol, 0.10 equiv). The resulting solution was stirred at 80° C. for 2 hours (under N2 atmosphere). The crude product was purified by preparative HPLC (conditions: XSelect CSH Prep C18 OBD Column, 5 μm, 19*150 mm; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 25 mL/minute; Gradient: 20% B to 55% B in 8 minutes; 254 nm; R_T: 7.12 minutes). Fractions containing the desired compound were evaporated to dryness to afford 7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-N-(1-methylpiperidin-4-yl)-4-oxo-4H,5H-thieno[3,2-c]pyridine-2-carboxamide (21 mg, 54%) as a white solid. ¹H NMR (400 MHz, Methanol-d4) δ 8.52 (s, 1H, FA), 8.26 (s, 1H), 7.86 (s, 1H), 7.07 (s, 2H), 4.40 (s, 2H), 4.02 (s, 7H), 3.75 (s, 3H), 3.27 (d, J=11.2 Hz, 2H), 2.90 (s, 6H), 2.79-2.67 (m, 2H), 2.62 (s, 3H), 2.12 (d, J=13.4 Hz, 2H), 1.94-1.79 (m, 2H). LCMS (ESI) m/z: [M+H]+=499.25.

Example 13—Preparation of 7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-2-(4-methylpiperazine-1-carbonyl)-4H,5H-thieno[3,2-c]pyridin-4-one (Compound B7)

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Step 1: Preparation of 7-bromo-5-methyl-2-(4-methylpiperazine-1-carbonyl)thieno[3,2-c]pyridin-4-one (i-23)

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To a solution of 7-bromo-5-methyl-4-oxothieno[3,2-c]pyridine-2-carboxylic acid (150.00 mg, 0.521 mmol, 1.00 equiv), HATU (296.93 mg, 0.781 mmol, 1.50 equiv) and DIEA (134.57 mg, 1.041 mmol, 2.00 equiv) in DMF (10.00 mL). The resulting mixture was stirred for 10 minutes at 25° C. Then 1-methylpiperazine (62.58 mg, 0.625 mmol, 1.20 equiv) was added to the reaction mixture. The resulting solution was stirred for 2 hours at 25° C. The resulting mixture was concentrated. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1:1). This resulted in 80 mg (42%) of 7-bromo-5-methyl-2-(4-methylpiperazine-1-carbonyl)thieno[3,2-c]pyridin-4-one as a yellow solid.

Step 2: Preparation of 7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-2-(4-methylpiperazine-1-carbonyl)-4H,5H-thieno[3,2-c]pyridin-4-one (Compound 87)

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To a solution of 7-bromo-5-methyl-2-(4-methylpiperazine-1-carbonyl)-4H,5H-thieno[3,2-c]pyridin-4-one (100.00 mg, 0.270 mmol, 1.00 equiv) and 052003 (263.99 mg, 0.810 mmol, 3.00 equiv) in 1,4-dioxane (5.00 mL) was added [4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]boronic acid (64.57 mg, 0.270 mmol, 1.00 equiv), Pd(dppf)Cl₂.CH₂Cl₂(22.06 mg, 0.027 mmol, 0.10 equiv), and H₂O (1.00 mL). The resulting solution was stirred for 2 hours at 80° C. The resulting solution was diluted with 10 mL of water. The resulting solution was extracted with ethyl acetate (3×10 mL) and the organic layers combined and dried over anhydrous sodium sulfate and concentrated. The crude product was purified by Prep-HPLC (conditions: 2 #SHIMADZU (HPLC-01); SunFire 018 OBD Prep Column, 19 mm×250 mm; mobile phase: Water (0.1% FA, formic acid) and ACN (hold 5% Phase B in 2 minutes, up to 12% in 10 minutes); UV Detector). This resulted in 5.3 mg (4.1%) of 7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-2-(4-methylpiperazine-1-carbonyl)-4H,5H-thieno[3,2-c]pyridin-4-one as a brown semi-solid. ¹H NMR (400 MHz, Methanol-d4) δ 8.45 (s, 2H, FA), 7.93 (s, 1H), 7.87 (s, 1H), 7.06 (s, 2H), 4.41 (s, 2H), 4.01 (s, 6H), 3.97-3.85 (m, 4H), 3.75 (s, 3H), 2.91 (s, 6H), 2.75-2.64 (m, 4H), 2.45 (s, 3H); LCMS (ESI) m/z: [M+H]+=485.20.

Example 14—Preparation of 2-(4-[2-[N-(1,1-dioxo-1λ6-thian-4-yl)carbamimidoyl]-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-7-yl]-2-methoxyphenoxy)acetic Acid Trifluoroacetic Acid (Compound B8 Trifluoroacetic Acid)

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Step 1 Preparation of 5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridine-2-carbonitrile (i-24)

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To a solution of 2-bromo-5-methyl-4H,5H-thieno[3,2-c]pyridin-4-one (500.00 mg, 2.048 mmol, 1.00 equiv) and Zn(CN)₂(529.22 mg, 4.506 mmol, 2.20 equiv) in DMF (20.00 mL) was added Pd(dppf)Cl₂(149.87 mg, 0.205 mmol, 0.10 equiv). The resulting mixture was stirred at 120° C. for 16 hours under N₂atmosphere. Fifteen identical reactions were set up in 15 sealed tubes in parallel. The reaction mixtures were combined and diluted with EA (500 mL). The resulting mixture was washed with water (3×300 mL) and saturated brine (300 mL). The organic layer was dried over Na₂SO₄, filtered, and evaporated to afford crude product. The crude product was purified by flash silica chromatography, elution gradient 0 to 100% EtOAc in petroleum ether. Pure fractions were evaporated to dryness to afford 5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridine-2-carbonitrile (4.4 g, 75%) as a black solid.

Step 2: Preparation of 7-bromo-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridine-2-carbonitrile (i-25)

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To a solution of 5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridine-2-carbonitrile (4.40 g, 23.131 mmol, 1.00 equiv) in THF (50.00 mL) was added NBS (6.18 g, 34.697 mmol, 1.50 equiv). The resulting mixture was stirred at room temperature for 16 hours. The reaction mixture was diluted with EA (400 mL). The resulting mixture was washed with water (3×200 mL) and saturated brine (300 mL). The organic layer was dried over Na₂SO₄, filtered, and evaporated to afford crude product. The crude product was purified by flash silica chromatography, elution gradient 0 to 100% EtOAc in petroleum ether. Pure fractions were evaporated to dryness to afford 7-bromo-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridine-2-carbonitrile (3.5 g, 56%) as a brown solid.

Step 3: Preparation of 7-bromo-N-(1,1-dioxo-1λ6-thian-4-yl)-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridine-2-carboximidamide (i-26)

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To a solution of 7-bromo-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridine-2-carbonitrile (3 g, 11.147 mmol, 1.00 equiv) in MeOH (45.00 mL, 11.451 mmol, 99.70 equiv) was added MeONa (2.00 g, 11.147 mmol, 1 equiv) solution in MeOH (30% by weight). The reaction mixture was heated to 75° C. for 3 hours. Then 4-amino-1λ6-thiane-1,1-dione hydrochloride (2.48 g, 0.013 mmol, 1.2 equiv) was added at 75° C., and the reaction mixture was stirred for a further 16 hours. The volatile components were removed in vacuo, and the resulting solid was suspended in MeOH (50 mL), filtered under reduced pressure, and washed with MeOH (20 mL) to afford 7-bromo-N-(1,1-dioxo-1λ6-thian-4-yl)-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridine-2-carboximidamide (3.0 g, 64%) as a brown solid.

Step 4: Preparation of Ethyl 2-(4-bromo-2-methoxyphenoxy)acetate (i-27)

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To a solution of 4-bromo-2-methoxyphenol (5 g, 24.626 mmol, 1 equiv) and K2003 (6.81 g, 49.253 mmol, 2 equiv) in acetone (50 mL) was added ethyl 2-bromoacetate (4.94 g, 29.580 mmol, 1.20 equiv). The resulting mixture was stirred at room temperature for 16 hours. The reaction mixture was diluted with EA (500 mL), The resulting mixture was washed with water (3×300 mL) and saturated brine (300 mL). The organic layer was dried over Na₂SO₄, filtered, and evaporated to afford crude product. The crude product was purified by flash silica chromatography, elution gradient 0 to 8% EtOAc in petroleum ether. Pure fractions were evaporated to dryness to afford ethyl 2-(4-bromo-2-methoxyphenoxy)acetate (4 g, 56%) as a yellow oil.

Step 5: Preparation of Ethyl 2-[2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy]acetate (i-28)

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To a solution of ethyl 2-(4-bromo-2-methoxyphenoxy)acetate (6 g, 20.752 mmol, 1 equiv) and 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (6.32 g, 24.903 mmol, 1.2 equiv) in solvent dioxane (120 mL) was added KOAc (4.07 g, 41.505 mmol, 2 equiv) and Pd(dppf)Cl₂(1.52 g, 2.075 mmol, 0.1 equiv). The resulting solution was stirred at 90° C. for 1 hour (under N2 atmosphere). The resulting mixture was washed with water (3×300 mL). The organic phase was concentrated under reduced pressure. The residue was purified by flash silica chromatography, elution gradient 0 to 60% EtOAc in petroleum ether. Pure fractions were evaporated to dryness to afford ethyl 2-[2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy]acetate (6.9 g, 98%) as a yellow oil.

Step 6: Preparation of ethyl 2-(4-[2-[N-(1,1-dioxo-1λ6-thian-4-yl)carbamimidoyl]-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-7-yl]-2-methoxyphenoxy)acetate (i-29)

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To a solution of 7-bromo-N-(1,1-dioxo-1λ6-thian-4-yl)-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridine-2-carboximidamide (1.00 g, 2.391 mmol, 1.00 equiv), ethyl 2-[2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy]acetate (0.88 g, 2.630 mmol, 1.10 equiv), and K3PO4 (1.01 g, 4.781 mmol, 2.00 equiv) in water (2.00 mL) and DMF (20.00 mL) was added Pd(dppf)Cl₂(0.17 g, 0.239 mmol, 0.10 equiv). The resulting mixture was stirred at 75° C. for 1 hour. The reaction mixture was diluted with EA (500 mL). The resulting mixture was washed with water (3×300 mL) and saturated brine (300 mL). The organic layer was dried over Na₂SO₄, filtered, and evaporated to afford crude product. The crude product was purified by flash silica chromatography, elution gradient 0 to 15% MeOH in DCM. Pure fractions were evaporated to dryness to afford ethyl 2-(4-[2-[N-(1,1-dioxo-1λ6-thian-4-yl)carbamimidoyl]-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-7-yl]-2-methoxyphenoxy)acetate (860 mg, 66%) as a brown solid.

Step 7: Preparation of 2-(4-[2-[N-(1,1-dioxo-1λ6-thian-4-yl)carbamimidoyl]-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-7-yl]-2-methoxyphenoxy)acetic Acid Trifluoroacetic Acid (Compound B8 Trifluoroacetic Acid)

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To a solution of ethyl 2-(4-[2-[N-(1,1-dioxo-1λ6-thian-4-yl)carbamimidoyl]-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-7-yl]-2-methoxyphenoxy)acetate (560 mg, 1.023 mmol, 1.00 equiv) in MeOH (25.00 mL) and H₂O (2.50 mL) was added LiOH.H₂O (429.48 mg, 10.226 mmol, 10.00 equiv). The resulting mixture was stirred at room temperature for 5 hours. The mixture was acidified to pH 2 with 1 N HCl (aq.). The crude product was purified by HPLC (conditions: XSelect CSH Prep C18 OBD Column, 5 μm, 19*150 mm; Mobile Phase A: Water (0.05% TFA), Mobile Phase B: ACN; Flow rate: 25 mL/minute; Gradient: 5% B to 22% B in 8 minutes; 254 nm; R_T: 5.68 minutes). Fractions containing the desired compound were evaporated to dryness to afford 2-(4-[2-[N-(1,1-dioxo-11A6-thian-4-yl)carbamimidoyl]-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-7-yl]-2-methoxyphenoxy)acetic acid trifluoroacetic acid (330 mg, 49%) as a yellow solid. LCMS (ES, m/z): 520.20. ¹H NMR (400 MHz, DMSO-d6) δ9.73 (d, J=8.1 Hz, 1H), 9.65 (s, 1H), 9.34 (s, 1H), 8.55 (s, 1H), 7.99 (s, 1H), 7.25-7.12 (m, 2H), 7.02 (d, J=8.4 Hz, 1H), 4.76 (s, 2H), 4.01 (d, J=8.9 Hz, 1H), 3.87 (s, 3H), 3.63 (s, 3H), 3.30-3.15 (m, 4H). 2.28-2.15 (m, 4H).

Example 15—Preparation of 7-(3,4-dimethoxyphenyl)-2-(1H-imidazol-2-yl)-5-methyl-4H,5H-thieno[3,2-c]pyridine-4-one (Compound B9)

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Step 1: Preparation of 7-bromo-2-(1H-imidazol-2-yl)-5-methyl-4H,5H-thieno[3,2-c]pyridin-4-one (i-30)

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To a solution of 7-bromo-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridine-2-carbonitrile (50.00 mg, 0.186 mmol, 1.00 equiv) and NaOMe (100.37 mg, 1.858 mmol, 10.00 equiv) in MeOH (1.00 mL) was stirred for 3 hours at room temperature. To the above mixture was added AcOH (446.28 mg, 7.432 mmol, 40.00 equiv) and HCl (1.50 mL, 49.368 mmol, 265.72 equiv) at room temperature. The resulting mixture was stirred for additional 16 hours at 75° C. The residue was dissolved in EtOAc (100 mL). The resulting mixture was washed with water (3×30 mL). The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (PE/EtOAc 5:1) to afford 7-bromo-2-(1H-imidazol-2-yl)-5-methyl-4H,5H-thieno[3,2-c]pyridin-4-one (30 mg, 52%) as a white solid. LCMS (ES, m/z): 310.0.

Step 2: Preparation of 7-(3,4-dimethoxyphenyl)-2-(1H-imidazol-2-yl)-5-methyl-4H,5H-thieno[3,2-c]pyridine-4-one (Compound 89)

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To a solution of 7-bromo-2-(1H-imidazol-2-yl)-5-methyl-4H,5H-thieno[3,2-c]pyridin-4-one (30.00 mg, 0.097 mmol, 1.00 equiv) and (3,4-dimethoxyphenyl)boronic acid (17.60 mg, 0.097 mmol, 1 equiv) in dioxane (1.50 mL) and H₂O (0.50 mL) was added Cs₂CO₃(63.03 mg, 0.193 mmol, 2.00 equiv) and XPhos-Pd-G3 (16.37 mg, 0.019 mmol, 0.2 equiv). The resulting solution was stirred at 80° C. for 1 hour (under N₂atmosphere). The crude product was purified by preparative HPLC (conditions: XBridge Shield RP18 OBD Column, 5 μm, 19*150 mm; Mobile Phase A: Water (10 mmol/L NH₄HCO₃), Mobile Phase B: ACN; Flow rate: 25 mL/minute; Gradient: 25% B to 33% B in 8 minutes; 254 nm; R_T: 6.77 minutes). Fractions containing the desired compound were evaporated to dryness to afford 7-(3,4-dimethoxyphenyl)-2-(1H-imidazol-2-yl)-5-methyl-4H,5H-thieno[3,2-c]pyridin-4-one (10.8 mg, 30%) as a white solid. LCMS (ES, m/z): 368.1. ¹H-NMR (400 MHz, CD₃OD) δ 7.96 (s, 1H), 7.62 (s, 1H), 7.26 (m, J=2.1 Hz, 1H), 7.24 (m, J=8.1, 2.2 Hz, 1H), 7.17 (s, 2H), 7.11 (m, J=8.1 Hz, 1H), 3.94 (s, 3H), 3.92 (s, 3H), 3.74 (s, 3H).

Example 16—Preparation of 7-(3,4-dimethoxyphenyl)-5-methyl-4H,5H-thieno[3,2-c]pyridin-4-one (Compound B10)

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Step 1: Preparation of 7-bromothieno[3,2-c]pyridin-4(5H)-one (i-32)

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A solution of 4H,5H-thieno[3,2-c]pyridin-4-one (1 g, 6.615 mmol, 1 equiv) and NBS (1.41 g, 7.938 mmol, 1.2 equiv) in DMF (10 mL) was stirred for 16 hours at room temperature. The resulting solution was diluted with of EtOAc. The resulting mixture was washed with water (3×50 mL). The resulting mixture was concentrated. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (30/70). Fractions containing the desired compound were evaporated to dryness to afford 7-bromo-4H,5H-thieno[3,2-c]pyridin-4-one (440.0 mg, 29%) as a purple solid. LCMS (ES, m/z): 230.0.

Step 2: Preparation of 7-bromo-5-methyl-4H,5H-thieno[3,2-c]pyridin-4-one (i-33)

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To a solution of 7-bromo-4H,5H-thieno[3,2-c]pyridin-4-one (200.00 mg, 0.869 mmol, 1.00 equiv) and Cs₂CO₃(849.67 mg, 2.608 mmol, 3 equiv) in THF (10.00 mL) was added MeI (246.76 mg, 1.739 mmol, 2 equiv) at room temperature. The resulting mixture was stirred for 2 hours at room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (hexane/EtOAc 5:1) to afford 7-bromo-5-methyl-4H,5H-thieno[3,2-c]pyridin-4-one (200 mg, 94%) as a brown solid. LCMS (ES, m/z): 244.0.

Step 3: Preparation of 7-(3,4-dimethoxyphenyl)-5-methyl-4H,5H-thieno[3,2-c]pyridin-4-one (Compound B10)

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To a solution of 7-bromo-5-methyl-4H,5H-thieno[3,2-c]pyridin-4-one (100.00 mg, 0.410 mmol, 1.00 equiv) and (3,4-dimethoxyphenyl)boronic acid (74.55 mg, 0.410 mmol, 1 equiv) in solvent H₂O (1.00 mL) and dioxane (3.00 mL) was added 052003 (266.95 mg, 0.819 mmol, 2 equiv) and XPhos-Pd-G3 (69.35 mg, 0.082 mmol, 0.2 equiv). The resulting solution was stirred at 80° C. for 1 hour (under N₂atmosphere). The crude product was purified by preparative HPLC (conditions: XSelect CSH Prep C18 OBD Column, 5 μm, 19*150 mm; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 25 mL/minute; Gradient: 20% B to 55% B in 8 minutes; 254 nm; R_T: 7.12 minutes). Fractions containing the desired compound were evaporated to dryness to afford 7-(3,4-dimethoxyphenyl)-5-methyl-4H,5H-thieno[3,2-c]pyridin-4-one (60.1 mg, 48%) as a white solid. LCMS (ES, m/z): 302. ¹H NMR (400 MHz, DMSO) δ 7.72 (s, 1H), 7.66 (d, J=5.4 Hz, 1H), 7.58 (d, J=5.4 Hz, 1H), 7.22-7.15 (m, 2H), 7.09 (d, J=8.1 Hz, 1H), 3.83 (d, J=6.4 Hz, 6H), 3.60 (s, 3H).

Example 17—Preparation of Intermediates
Preparation of Ethyl 2-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-(trifluoromethyl)phenoxy]-acetate (i-35)

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Step 1: Preparation of Ethyl 2-[4-bromo-2-(trifluoromethyl)phenoxy]acetate (i-34)

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To a solution of 4-bromo-2-(trifluoromethyl)phenol (5 g, 20.746 mmol, 1 equiv) and K₂CO₃(5.73 g, 41.493 mmol, 2 equiv) in acetone (50 mL) was added ethyl 2-bromoacetate (5.20 g, 31.119 mmol, 1.5 equiv) at room temperature. The resulting mixture was stirred for 1 hour at room temperature. The solid was then filtered off, the filtrate was concentrated, and the residue was purified by silica gel column chromatography, eluted with PE/EtOAc (5:1) to afford ethyl 2-[4-bromo-2-(trifluoromethyl)phenoxy] acetate (6.5 g, 96%) as a white solid. LCMS (ESI) m/z: [M+H]+=327.

Step 2: Preparation of Ethyl 2-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-(trifluoromethyl)phenoxy]-acetate (i-35)

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To a solution of ethyl 2-[4-bromo-2-(trifluoromethyl)phenoxy]acetate (1 g, 3.057 mmol, 1 equiv) and 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (931.6 mg, 3.669 mmol, 1.2 equiv) in dioxane (10 mL) was added KOAc (0.60 g, 6.114 mmol, 2.00 equiv) and Pd(dppf)Cl₂(223.7 mg, 0.306 mmol, 0.1 equiv). The resulting mixture was stirred for 1 h at 90° C. under a nitrogen atmosphere. The mixture was concentrated and the residue was purified by silica gel column chromatography, eluted with PE/EtOAc (5:1) to afford ethyl 2-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-(trifluoromethyl)phenoxy]acetate (800 mg, 70%) as a yellow oil. LCMS (ESI) m/z: [M+H]+=375.

Preparation of Tert-Butyl N-(8-[[2-(2,6-dioxopiperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-4-yl]amino]octyl)carbamate (i-37)

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Step 1: Preparation of Tert-Butyl (8-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4yl)amino)octyl)carbamate (i-36)

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To a solution of 3-(4-amino-1-oxo-2,3-dihydro-1H-isoindol-2-yl)piperidine-2,6-dione (200 mg, 0.773 mmol, 1 equiv) and tert-butyl N-(8-oxooctyl)carbamate (225.31 mg, 0.927 mmol, 1.2 equiv) in MeOH (10 mL) was added NaBH₃CN (145.41 mg, 2.319 mmol, 3 equiv) and AcOH (0.5 mL). The resulting solution was stirred at 25° C. for 1 hour. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (5:1) to afford tert-butyl N-(8-[[2-(2,6-dioxopiperidin-3-yl)-1-oxo-2,3-ihydro-1H-isoindol-4-yl]amino]octyl)carbamate (202 mg, 62%) as an off-white solid. LCMS (ESI) m/z: [M+H]+=487.

Step 2: Preparation of tert-butyl N-(8-[[2-(2,6-dioxopiperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-4-yl]amino]octyl)carbamate (i-37)

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To a solution of tert-butyl N-(8-[[2-(2,6-dioxopiperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-4-yl]amino]octyl)carbamate (100 mg, 0.310 mmol, 1 equiv) in DCM (5.0 ml) was added TFA (2.0 ml). The resulting solution was stirred at 25° C. for 1 hour. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography (conditions: column, 018 silica gel; mobile phase, MeCN in water, 10% to 50% gradient in 10 minutes; detector, UV 254 nm). This resulted in 3-[4-[(8-aminooctyl)amino]-1-oxo-2,3-dihydro-1H-isoindol-2-yl]piperidine-2,6-dione (103 mg, 87%) as a yellow solid. LCMS (ESI) m/z: [M+H]+=387.

Preparation of 5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)pentanal (i-39)

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Step 1: Preparation of 4-(4-(1,3-dioxolan-2-yl)butoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (i-38)

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To a stirred mixture of 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindole-1,3-dione (400 mg, 1.459 mmol, 1.00 equiv) and 2-(4-bromobutyl)-1,3-dioxolane (274.5 mg, 1.313 mmol, 0.90 equiv) in DMF (4.00 mL) was added Cs₂CO₃(950.5 mg, 2.917 mmol, 2.00 equiv) at 60° C. under nitrogen atmosphere. The resulting mixture was stirred for 5 hours. The resulting mixture was filtered with acetonitrile (200 mL), concentrated, and purified by Prep-TLC (PE/EtOAc (1:1)) to afford 4-[4-(1,3-dioxolan-2-yl)butoxy]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (81 mg, 13%) as a colorless oil. LCMS (ESI) m/z: [M+H]⁺=403.

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

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To a stirred mixture of 4-[4-(1,3-dioxolan-2-yl)butoxy]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (46.0 mg, 0.114 mmol, 1.00 equiv) in H₂O (3.0 mL) was added 4 M HCl in dioxane (3 mL) dropwise at room temperature under nitrogen atmosphere. After 3 hours, saturated NaHCO₃solution (50 mL) was added to the mixture. The mixture was then extracted with EtOAc (100 mL×3), and the combined organic layers were concentrated to give 5-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]pentanal (35 mg, 70%) as a light yellow semi-solid. LCMS (ESI) m/z: [M+H]₊=359.

Preparation of tert-butyl 4-([7-chloro-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl]carbamoyl)piperidine-1-carboxylate (i-40)

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To a solution of 2-amino-7-chloro-5-methylthieno[3,2-c]pyridin-4-one (300.0 mg, 1.397 mmol, 1.00 equiv)v) and 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (416.5 mg, 1.817 mmol, 1.3 equiv) in solvent DMF (4 mL) was added DIEA (722.5 mg, 5.590 mmol, 4 equiv) and HATU (1062.7 mg, 2.795 mmol, 2 equiv). The resulting solution was stirred at 25° C. for 2 hours. The mixture was diluted with EtOAc (30 mL) and washed with water (30 mL×3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. The crude product was purified by chromatography on silica gel eluted with DCM/MeOH (from 20:1 to 10:1) to give tert-butyl 4-([7-chloro-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl]carbamoyl)piperidine-1-carboxylate (360 mg, 61%) as a brown solid. LCMS (ESI) m/z: [M+H]+=426.

The following compounds (intermediates i-41, i-42, and i-43) were prepared in a similar manner as described in the preparation of intermediate i-40.

Compound (Intermediate No.)
Structure
Analytical Data

tert-butyl 3-([7-chloro-5-methyl-4-oxothieno[3,2- c]pyridin-2-yl]carbamoyl)piperidine-1- carboxylate (i-41)

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340 mg (57%) as a yellow oil; LCMS (ESI) m/z: [M + H]⁺ = 426.12

7-chloro-5-methyl-4-oxothieno[3,2-c]pyridin-2- yl]-1-methylpiperidine-4-carboxamide (i-42)

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80 mg (51%) as a yellow solid; LCMS (ESI) m/z: [M + H]+ = 340

tert-butyl 4-(7-bromo-5-methyl-4-oxo-4,5- dihydrothieno[3,2-c]pyridine-2- carboxamido)piperidine-1-carboxylate (i-42)

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154 mg (31%) as a yellow solid; LCMS (ESI) m/z: [M + H]⁺ = 470

Preparation of tert-butyl 2-[([2,6-dimethoxy-4-[5-methyl-2-(1-methylpiperidine-4-amido)-4-oxothieno[3,2-c]pyridin-7-yl]phenyl]methyl)(methyl)amino]acetate (i-44)

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To a solution of N-[7-chloro-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl]-1-methylpiperidine-4-carboxamide (50.0 mg, 0.147 mmol, 1.00 equiv) and 4-([[2-(tert-butoxy)-2-oxoethyl](methyl)amino]methyl)-3,5-dimethoxyphenylboronic acid (59.9 mg, 0.177 mmol, 1.20 equiv) in solvent dioxane (1.50 mL) and H₂O (0.50 mL) was added Pd(dppf)Cl₂*CH₂Cl₂(8.5 mg, 0.015 mmol, 0.10 equiv) and 052003 (95.9 mg, 0.294 mmol, 2.00 equiv), and the resulting solution was stirred at 90° C. for 3 hours. The reaction mixture was then diluted with 10 mL of water and extracted ethyl acetate (2×20 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was applied onto a silica gel column with CH₂Cl₂/MeOH (20:1). This resulted in tert-butyl 2-[([2,6-dimethoxy-4-[5-methyl-2-(1-methylpiperidine-4-amido)-4-oxothieno[3,2-c]pyridin-7-yl]phenyl]methyl)(methyl)amino]acetate (40 mg, 45%) as a yellow solid. LCMS (ESI) m/z: [M+H]+=599.

The following compounds (intermediates i-45, i-46, and i-47) were prepared in a similar manner as described in the preparation of intermediate i-44.

Compound (Intermediate No.)
Structure
Analytical Data

tert-butyl 4-[(7-[4-[(dimethylamino)methyl]-3,5- dimethoxyphenyl]-5-methyl-4-oxothieno[3,2- c]pyridin-2-yl)carbamoyl]piperidine-1- carboxylate (i-45)

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168 mg (34%) as a brown solid; LCMS (ESI) m/z: [M + H]+ = 585

tert-butyl 3-[(7-[4-[(dimethylamino)methyl]-3,5- dimethoxyphenyl]-5-methyl-4-oxothieno[3,2- c]pyridin-2-yl)carbamoyl]piperidine-1- carboxylate (i-46)

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260 mg (63%) as a red oil; LCMS (ESI) m/z: [M + H]+ = 585.27

tert-butyl 4-(7-(4-((dimethylamino)methyl)-3,5- dimethoxyphenyl)-5-methyl-4-oxo-4,5- dihydrothieno[3,2-c]pyridine-2- carboxamido)piperidine-1-carboxylate (i-47)

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122 mg (62%) as a brown solid; LCMS (ESI) m/z: [M + H]⁺ = 585

Preparation of ethyl 2-(4-[2-[N-(1,1-dioxo-1λ6-thian-4-yl)carbamimidoyl]-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-7-yl]-2-(trifluoromethyl)phenoxy)acetate (i-48)

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To a solution of ethyl 2-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-(trifluoromethyl) phenoxy]acetate (100 mg, 0.267 mmol, 1 equiv) and 7-bromo-N-(1,1-dioxo-1λ6-thian-4-yl)-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridine-2-carboximidamide (111.8 mg, 0.267 mmol, 1 equiv) in DMF (3 mL) and H₂O (0.3 mL) was added K3PO4 (113.46 mg, 0.535 mmol, 2 equiv) and Pd(dppf)Cl₂(39.1 mg, 0.053 mmol, 0.2 equiv) at room temperature. The resulting mixture was stirred for 1 hour at 75° C. under nitrogen atmosphere. The mixture was concentrated and the residue was purified by silica gel column chromatography, eluted with CH₂Cl₂/MeOH (5:1) to afford ethyl 2-(4-[2-[N-(1,1-dioxo-1λ6-thian-4-yl)carbamimidoyl]-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-7-yl]-2-(trifluoromethyl)phenoxy) acetate (97 mg, 62%) as an off-white solid. LCMS (ESI) m/z: [M+H]+=586.

Preparation of N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl)piperidine-4-carboxamide (i-49)

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To a solution of tert-butyl 4-[(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl)carbamoyl]piperidine-1-carboxylate (168.0 mg, 0.287 mmol, 1.00 equiv) in

DCM (9.00 mL) was added TFA (3.00 mL, 13.463 mmol, 262.41 equiv), and the resulting solution was stirred at 25° C. for 2 hours. The solvent was removed to afford N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl)piperidine-4-carboxamide (180 mg, crude) as a brown solid that was used directly without further purification. LCMS (ESI) m/z: [M+H]+=485.

The following compounds (intermediates i-50 and i-51) were prepared in a similar manner as described in the preparation of intermediate i-49.

Compound (Intermediate No.)
Structure
Analytical Data

N-(7-[4-[(dimethylamino)methyl]-3,5- dimethoxyphenyl]-5-methyl-4- oxothieno[3,2-c]pyridin-2-yl)piperidine-3- carboxamide (i-50)

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160 mg (88%) as a red oil; LCMS (ESI) m/z: [M + H]⁺ = 485.21

N-(7-[4-[(dimethylamino)methyl]-3,5- dimethoxyphenyl]-5-methyl-4- oxothieno[3,2-c]pyridin-2-yl)piperidine-4- carboxamide (i-51)

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180 mg (crude) as a brown solid (used directly without further purification); LCMS (ESI) m/z: [M + H]+ = 485

Preparation of [([2,6-dimethoxy-4-[5-methyl-2-(1-methylpiperidine-4-amido)-4-oxothieno[3,2-c]pyridin-7-yl]phenyl]methyl)(methyl)amino]acetic Acid (i-52)

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To a solution of tert-butyl 2-[([2,6-dimethoxy-4-[5-methyl-2-(1-methylpiperidine-4-amido)-4-oxothieno[3,2-c]pyridin-7-yl]phenyl]methyl)(methyl)amino]acetate (40.0 mg, 0.067 mmol, 1.00 equiv) in DCM (2.00 mL) was added TFA (2.00 mL), and the resulting solution was stirred at 25° C. for 1 hour. The resulting mixture was concentrated to afford [([2,6-dimethoxy-4-[5-methyl-2-(1-methylpiperidine-4-amido)-4-oxothieno[3,2-c]pyridin-7-yl]phenyl]methyl)(methyl)amino]acetic acid (50 mg, crude) as a yellow solid that was used directly without further purification.

Preparation of 2-(4-[2-[N-(1,1-dioxo-1λ6-thian-4-yl)carbamimidoyl]-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-7-yl]-2-(trifluoromethyl)phenoxy)acetic Acid (i-53)

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A solution of ethyl 2-(4-[2-[N-(1,1-dioxo-1λ6-thian-4-yl)carbamimidoyl]-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-7-yl]-2-(trifluoromethyl)phenoxy)acetate (90 mg, 0.154 mmol, 1 equiv) and LiOH.H₂O (64.49 mg, 1.537 mmol, 10 equiv) in MeOH (2 mL) and H₂O (0.5 mL) was stirred for 1 hour at room temperature. The resulting mixture was concentrated under vacuum and afforded 2-(4-[2-[N-(1,1-dioxo-1A6-thian-4-yl)carbamimidoyl]-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-7-yl]-2-(trifluoromethyl)phenoxy)acetic acid (80 mg, 93%) as a yellow solid. LCMS (ESI) m/z: [M+H]+=558.

Preparation of tert-butyl 4-[7-bromo-5-methyl-4-oxothieno[3,2-c]pyridine-2-amido]-3,3-difluoro piperidine-1-carboxylate (i-55)

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To a solution of 7-bromo-5-methyl-4-oxothieno[3,2-c]pyridine-2-carboxylic acid (200.0 mg, 0.694 mmol, 1.00 equiv) and tert-butyl 4-amino-3,3-difluoropiperidine-1-carboxylate (213.2 mg, 0.902 mmol, 1.3 equiv) in DMF (4 mL) was added DIEA (448.57 mg, 3.471 mmol, 5 equiv) and HATU (527.8 mg, 1.388 mmol, 2 equiv), and the resulting solution was stirred at room temperature for 4 hours. The mixture was diluted with EtOAc (20 mL) and washed with water (30 mL×3). The organic layers were combined and dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. The crude product was purified by chromatography on silica gel eluted with DCM/MeOH (from 30:1 to 20:1) to give tert-butyl 4-[7-bromo-5-methyl-4-oxothieno[3,2-c]pyridine-2-amido]-3,3-difluoropiperidine-1-carboxylate (100 mg, 28.45%) as a brown yellow solid. LCMS (ESI) m/z: [M+H]+=506.

Preparation of Tert-Butyl 4-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridine-2-amido)-3,3-difluoropiperidine-1-carboxylate (i-57)

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To a solution of tert-butyl 4-[7-bromo-5-methyl-4-oxothieno[3,2-c]pyridine-2-amido]-3,3-difluoropiperidine-1-carboxylate (90.0 mg, 0.178 mmol, 1.00 equiv) and 4-[(dimethylamino)methyl]-3,5-dimethoxyphenylboronic acid (63.7 mg, 0.267 mmol, 1.5 equiv) in dioxane (4 mL) and H₂O (0.4 mL) was added 052003 (115.8 mg, 0.355 mmol, 2 equiv) and RuPhos Palladacycle Gen.4 (15.1 mg, 0.018 mmol, 0.1 equiv). The resulting solution was stirred at room temperature for 2 hours (under N₂atmosphere). The mixture was diluted with EtOAc (20 mL) and washed with water (30 mL×3). The organic layers were combined and dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. The crude product was purified by chromatography on silica gel eluted with DCM/MeOH (from 20:1 to 10:1) to give tert-butyl 4-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridine-2-amido)-3,3-difluoropiperidine-1-carboxylate (66 mg, 59.82%) as an off-white solid. LCMS (ESI) m/z: [M+H]+=621.

Preparation of N-(3,3-difluoropiperidin-4-yl)-7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridine-2-carboxamide (i-58)

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To a solution of tert-butyl 4-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridine-2-amido)-3,3-difluoropiperidine-1-carboxylate (66.0 mg, 0.106 mmol, 1.00 equiv) in DCM (9.00 mL) was added TFA (3 mL, 40.389 mmol, 379.85 equiv). The resulting solution was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure to afford N-(3,3-difluoropiperidin-4-yl)-7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridine-2-carboxamide (66 mg, crude) as a yellow solid that was used directly without further purification. LCMS (ESI) m/z: [M+H]+=521.

Preparation of 7-bromo-5-methyl-N-(1-methylpiperidin-4-yl)-4-oxo-4,5-dihydrothieno[3,2-c]pyridine-2-carboxamide (i-59)

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To a stirred mixture of 7-bromo-5-methyl-4-oxothieno[3,2-c]pyridine-2-carboxylic acid (200.00 mg, 0.694 mmol, 1.00 equiv) and 1-methylpiperidin-4-amine (118.90 mg, 1.041 mmol, 1.50 equiv) in DMF (2.00 mL) was added HATU (527.88 mg, 1.388 mmol, 2 equiv) at 0° C. After 10 minutes, DIEA (358.86 mg, 2.777 mmol, 4.00 equiv) was added, and the reaction mixture was stirred for 10 minutes at 0° C. The mixture was allowed to stirred one hour at room temperature. The resulting mixture was extracted with the mixture of DCM and isopropanol (3:1; 3×100 mL), concentrated, and purified by silica gel column chromatography, eluted with DCM/MeOH (10:1) to afford 7-bromo-5-methyl-N-(1-methylpiperidin-4-yl)-4-oxothieno[3,2-c]pyridine-2-carboxamide (232 mg, 82.10%) as a yellow solid. LCMS (ESI) m/z: [M+H]⁺=384.

Preparation of 7-(4-formyl-3,5-dimethoxyphenyl)-5-methyl-N-(1-methylpiperidin-4-yl)-4-oxo-4,5-dihydrothieno[3,2-c]pyridine-2-carboxamide (i-60)

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To a stirred mixture of 7-bromo-5-methyl-N-(1-methylpiperidin-4-yl)-4-oxothieno[3,2-c]pyridine-2-carboxamide (232.00 mg, 0.604 mmol, 1.00 equiv) and 4-formyl-3,5-dimethoxyphenylboronic acid (190.16 mg, 0.906 mmol, 1.50 equiv) in dioxane (4.00 mL) and H₂O (0.80 mL) was added Pd(dppf)Cl₂.CH₂Cl₂(49.30 mg, 0.060 mmol, 0.10 equiv) and Cs₂CO₃(590.10 mg, 1.811 mmol, 3.00 equiv). The mixture was allowed to stir at 80° C. under nitrogen atmosphere for 3 hours. The resulting mixture was filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with DCM/MeOH (10:1) to afford 7-(4-formyl-3,5-dimethoxyphenyl)-5-methyl-N-(1-methylpiperidin-4-yl)-4-oxothieno[3,2-c]pyridine-2-carboxamide (131 mg, 43.67%) as a yellow solid. LCMS (ESI) m/z: [M+H]⁺=470.

Preparation of tert-butyl 2-[[(2,6-dimethoxy-4-[5-methyl-2-[(1-methylpiperidin-4-yl)carbamoyl]-4-oxothieno[3,2-c]pyridin-7-yl]phenyl)methyl]methyl)amino]acetate (i-61)

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To a stirred mixture of 7-(4-formyl-3,5-dimethoxyphenyl)-5-methyl-N-(1-methylpiperidin-4-yl)-4-oxothieno[3,2-c]pyridine-2-carboxamide (131.00 mg, 0.279 mmol, 1.00 equiv) and tert-butyl 2-(methylamino)acetate hydrochloride (60.82 mg, 0.335 mmol, 1.20 equiv) in MeOH (1.50 mL) was added TEA (84.69 mg, 0.837 mmol, 3.00 equiv) at room temperature. After 30 minutes, NaBH₃CN (52.60 mg, 0.837 mmol, 3.00 equiv) was added at room temperature. After 3 hours, the reaction mixture was purified by silica gel column chromatography, eluted with DCM/MeOH (20:1) to afford tert-butyl 2-[[(2,6-dimethoxy-4-[5-methyl-2-[(1-methylpiperidin-4-yl)carbamoyl]-4-oxothieno[3,2-c]pyridin-7-yl]phenyl)methyl] (methyl)amino]acetate (84 mg, 50.28%) as a yellow solid. LCMS (ESI) m/z: [M+H]⁺=599.

Preparation of [[(2,6-dimethoxy-4-[5-methyl-2-[(1-methylpiperidin-4-yl)carbamoyl]-4-oxothieno[3,2-c]pyridin-7-yl]phenyl)methyl]methyl)amino] trifluoroacetic Acid (i-62)

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To a stirred mixture of tert-butyl 2-[[(2,6-dimethoxy-4-[5-methyl-2-[(1-methylpiperidin-4-yl)carbamoyl]-4-oxothieno[3,2-c]pyridin-7-yl]phenyl)methyl](methyl)amino]acetate (84.00 mg, 0.140 mmol, 1.00 equiv) in DCM (4.00 mL) was added TFA (1.00 mL) dropwise at room temperature and stirred for one hour at room temperature. The resulting mixture was concentrated under vacuum. This resulted in [[(2,6-dimethoxy-4-[5-methyl-2-[(1-methylpiperidin-4-yl)carbamoyl]-4-oxothieno[3,2-c]pyridin-7yl]phenyl) methyl](methyl) amino] trifluoroacetic acid (101 mg, 96.48%) as a yellow crude solid. LCMS (ESI) m/z: [M+H]⁺=543.

Example 18—Preparation of N-[7-(4-[[([[2-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]ethoxy)ethyl]carbamoyl]methyl)(methyl)amino]methyl]-3,5-dimethoxyphenyl)-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl]-1-methylpiperidine-4-carboxamide Formic Acid (Compound D1 Formic Acid)

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To a solution of [([2,6-dimethoxy-4-[5-methyl-2-(1-methylpiperidine-4-amido)-4-oxothieno[3,2-c]pyridin-7-yl]phenyl]methyl)(methyl)amino]acetic acid (50.0 mg, 0.092 mmol, 1.00 equiv) and HATU (70.1 mg, 0.184 mmol, 2.00 equiv) in DMF (2.00 mL) was added 4-[2-(2-aminoethoxy)ethoxy]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (33.3 mg, 0.092 mmol, 1.00 equiv) and DIEA (35.7 mg, 0.276 mmol, 3.00 equiv). The resulting solution was stirred at 25° C. for 2 hours. The crude product was purified by preparative HPLC (conditions: XSelect CSH Prep C18 OBD Column, 5 μm, 19*150 mm; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 25 mL/minute; Gradient: 20% B to 55% B in 8 minutes; 254 nm; R_T: 7.12 minutes) to afford N-[7-(4-[[([[2-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]ethoxy)ethyl]carbamoyl]methyl)(methyl)amino]methyl]-3,5-dimethoxyphenyl)-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl]-1-methylpiperidine-4-carboxamide formic acid (7 mg, 8.6%) of as a yellow solid. ¹H NMR (300 MHz, Methanol-d4) δ 8.40 (br s, 0.4H, FA), 7.64-7.53 (m, 2H), 7.25 (d, J=7.2 Hz, 1H), 7.20-7.09 (m, 2H), 6.97 (s, 2H), 5.07 (dd, J=12.9, 5.3 Hz, 1H), 4.29 (s, 2H), 4.07-3.90 (m, 8H), 3.73 (s, 4H), 3.63-3.48 (m, 7H), 3.46-3.38 (m, 2H), 3.18-3.02 (m, 2H), 2.91-2.72 (m, 9H), 2.64-2.47 (m, 1H), 2.25-1.98 (m, 5H). LCMS (ESI) m/z: [M+H]+=886.80.

The following compound (compound D2) was prepared in a similar manner as described for compound D1.

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2-(4-[2-[N-(1,1-dioxo-A6-thian-4-yl) carbamimidoyl]-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-7-yl]-2-methoxyphenoxy)-N-(8-[[2-(2,6-dioxopiperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-4-yl]amino]octyl)acetamide (14 mg, 16%) as a yellow solid. ¹H NMR (300 MHz, Methanol-d4) δ 8.54 (s, 0.4H, FA), 8.21 (s, 1H), 7.74 (s, 1H), 7.34-7.16 (m, 3H), 7.13 (d, J=8.3 Hz, 1H), 7.02 (d, J=7.5 Hz, 1H), 6.75 (d, J=8.0 Hz, 1H), 5.16 (dd, J=13.3, 5.1 Hz, 1H), 4.59 (d, J=5.2 Hz, 3H), 4.28 (d, J=3.0 Hz, 2H), 3.96 (s, 3H), 3.73 (s, 3H), 3.29-3.10 (m, 4H), 3.03-2.79 (m, 3H), 2.51 (td, J=13.2, 5.0 Hz, 1H), 2.32-2.15 (m, 5H), 1.61 (dd, J=15.5, 7.3 Hz, 4H), 1.36 (s, 8H). LCMS (ESI) m/z: [M+H]+=888.30.

Example 19—Preparation of 2-(4-[2-[N-(1,1-dioxo-1λ6-thian-4-yl)carbamimidoyl]-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-7-yl]-2-(trifluoromethyl)phenoxy)-N-(8-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-4-yl]amino]octyl)acetamide (Compound D3)

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To a solution of 2-(4-[2-[N-(1,1-dioxo-1λ6-thian-4-yl)carbamimidoyl]-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-7-yl]-2-(trifluoromethyl)phenoxy)acetic acid (69 mg, 0.124 mmol, 1 equiv) and DIEA (N,N-diisopropylamine) (48 mg, 0.371 mmol, 3 equiv) in DMF (1 mL) was added PyBOP (96.6 mg, 0.186 mmol, 1.5 equiv) and 4-[(8-aminooctyl)amino]-2-(2,6-dioxopiperidin-3-yl)-2,3-dihydro-1H-isoindole-1,3-dione (49.6 mg, 0.124 mmol, 1 equiv). The resulting solution was stirred at room temperature for overnight. Without workup, the crude product (69 mg) was purified by Prep-HPLC (conditions: SunFire Prep 018 OBD Column 19×150 mm 5 μm 10 nm; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 25 mL/minute; Gradient: 25% B to 55% B in 8 minutes; 254 nm; R_T: 7.35 minutes) to afford 2-(4-[2-[N-(1,1-dioxo-1λ6-thian-4-yl)carbamimidoyl]-5-methyl-4-oxo-4H,5H-thieno[3,2-c]pyridin-7-yl]-2-(trifluoromethyl)phenoxy)-N-(8-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-4-yl]amino]octyl)acetamide (30 mg, 25%) as a yellow solid. ¹H NMR (300 MHz, DMSO-d6) δ 11.10 (s, 1H), 8.23 (s, 1H), 8.14 (s, 0.26H, FA), 7.94-7.78 (m, 4H), 7.62-7.50 (m, 1H), 7.30 (d, J=8.8 Hz, 1H), 7.03 (dd, J=13.8, 7.8 Hz, 2H), 6.51 (t, J=5.9 Hz, 1H), 5.05 (dd, J=12.8, 5.3 Hz, 1H), 4.74 (s, 2H), 3.69 (s, 1H), 3.58 (s, 3H), 3.26 (d, J=6.6 Hz, 2H), 3.20-3.09 (m, 5H), 3.12-2.96 (m, 1H), 2.98-2.79 (m, 1H), 2.61 (s, 1H), 2.03 (s, 4H), 1.79-1.68 (m, 1H), 1.55-1.45 (m, 4H), 1.30 (s, 8H). LCMS (ESI) m/z: [M+H]+=940.50.

Example 20—Preparation of N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl)-1-[3-[2-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]amino]ethoxy)ethoxy]propanoyl]piperidine-4-carboxamide Formic Acid (Compound D4)

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To a solution of 3-[2-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]amino]ethoxy)ethoxy]propanoic acid (39.4 mg, 0.091 mmol, 1.1 equiv), HOBt (22.3 mg, 0.165 mmol, 2 equiv), and EDCI (31.7 mg, 0.165 mmol, 2 equiv) in DMF (1 mL) was added N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl)piperidine-4-carboxamide (40.0 mg, 0.083 mmol, 1.00 equiv) and DIEA (53.4 mg, 0.413 mmol, 5 equiv), and the resulting solution was stirred at 25° C. for 2 hours. Without any additional work-up, the mixture was purified by Prep-HPLC (conditions: SunFire 018 OBD Prep Column, 100 Å, 5 μm, 19 mm×250 mm; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate:25 mL/minute; Gradient:11 B to 26 B in 13 minutes; 254 nm) to give N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl)-1-[3-[2-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]amino]ethoxy)ethoxy]propanoyl]piperidine-4-carboxamide formic acid (22.3 mg, 29%). ¹H NMR (400 MHz, Methanol-d4) δ 8.56 (br s, 0.8H, FA), 7.64 (s, 1H), 7.47-7.41 (m, 1H), 7.15 (d, J=2.2 Hz, 1H), 7.03-6.98 (m, 3H), 6.95 (d, J=7.2 Hz, 1H), 5.02 (dd, J=12.7, 5.4 Hz, 1H), 4.59 (d, J=13.3 Hz, 1H), 4.28 (s, 2H), 4.13 (d, J=13.9 Hz, 1H), 3.99 (s, 6H), 3.81-3.66 (m, 9H), 3.65-3.61 (m, 2H), 3.52-3.44 (m, 2H), 3.23-3.14 (m, 1H), 2.88-2.79 (m, 8H), 2.77-2.60 (m, 4H), 2.55-2.46 (m, 1H), 2.11-2.01 (m, 1H), 1.95-1.74 (m, 3H), 1.73-1.60 (m, 1H). LCMS (ESI) m/z: [M+H]+=900.50.

Example 21—Preparation of N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl)-1-[3-[2-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]amino]ethoxy)ethoxy]propanoyl]piperidine-3-carboxamide (Compound D5)

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To a stirred solution of 3-[2-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]amino]ethoxy)ethoxy] propanoic acid (53.66 mg, 0.124 mmol, 1.20 equiv) in DMF (1.00 mL) was added HOBT (27.88 mg, 0.206 mmol, 2.00 equiv) and EDCI (39.56 mg, 0.206 mmol, 2.00 equiv) at 25° C. The resulting mixture was stirred for 20 minutes at 25° C. Then N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl)piperidine-3-carboxamide (50.00 mg, 0.103 mmol, 1.00 equiv) and DIEA (40.00 mg, 0.310 mmol, 3.00 equiv) was added to the reaction mixture. The resulting mixture was stirred for 2 hours at room temperature. The crude product was purified by Prep-HPLC (conditions: SunFire 018 OBD Prep Column, 100 Å, 5 μm, 19 mm×250 mm; mobile phase, Water (0.1% FA) and ACN (11% Phase B up to 23% in 18 minutes); Detector, UV) to afford N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl)-1-[3-[2-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]amino]ethoxy)ethoxy]propanoyl]piperidin e-3-carboxamide (12 mg, 13%) as a green solid. ¹H NMR (400 MHz, Methanol-d4) δ 8.57 (brs, 0.8H, FA), 7.66-7.58 (m, 1H), 7.57-7.25 (m, 1H), 7.19-7.06 (m, 1H), 7.05-6.95 (m, 3H), 6.86-6.76 (m, 1H), 5.04 (dd, J=12.5, 5.4 Hz, 1H), 4.53-4.40 (m, 1H), 4.37-4.24 (m, 0.5H), 4.20 (d, J=7.0 Hz, 2H), 4.03-3.98 (m, 6H), 3.95-3.88 (m, 0.5H), 3.81-3.62 (m, 11H), 3.52-3.48 (m, 1H), 3.45-3.36 (m, 1.5H), 3.21-3.12 (m, 0.5H), 3.06-2.92 (m, 1H), 2.87-2.63 (m, 11H), 2.61-2.53 (m, 1H), 2.25-1.98 (m, 2H), 1.86-1.73 (m, 2H), 1.69-1.46 (m, 1H). LCMS (ESI) m/z: [M+H]+=900.50.

Example 22—Preparation of N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl)-1-(5-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]pentyl)piperidine-4-carboxamide Formic Acid (Compound D6 Formic Acid)

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To a solution of N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl)piperidine-4-carboxamide (30.0 mg, 0.062 mmol, 1.00 equiv) and 5-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]pentanal (24.4 mg, 0.068 mmol, 1.10 equiv) in DMF (1.00 mL) was added sodium triacetoxyborohydride (26.2 mg, 0.124 mmol, 2.00 equiv), and the resulting solution was stirred at room temperature for 2 hours. Without any additional work-up, the mixture was purified by Prep-HPLC (conditions: Xselect CSH F-Phenyl OBD column, 19*250, 5 μm; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 25 mL/minute; Gradient:7 B to 19 B in 12 minutes; 254/220 nm) to give N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl)-1-(5-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]pentyl)piperidine-4-carboxamide; formic acid (4.7 mg, 8.7%). ¹H NMR (400 MHz, Methanol-d4) δ 8.55 (br s, 1.1H, FA), 7.67 (s, 1H), 7.63-7.57 (m, 1H), 7.29 (d, J=7.1 Hz, 1H), 7.21 (s, 1H), 7.16 (d, J=8.4 Hz, 1H), 7.05 (s, 2H), 5.13 (dd, J=12.9, 5.5 Hz, 1H), 4.38 (s, 2H), 4.02 (s, 6H), 3.85 (t, J=7.0 Hz, 2H), 3.74 (s, 3H), 3.39-3.35 (m, 2H), 2.93-2.87 (m, 8H), 2.78 (s, 2H), 2.71-2.56 (m, 4H), 2.15-2.08 (m, 1H), 2.05-1.92 (m, 4H), 1.77-1.60 (m, 4H), 1.44-1.34 (m, 2H). LCMS (ESI) m/z: [M+H]+=827.50.

Compound D11 was prepared in a similar manner as described for compound D6.

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7-(4-((dimethylamino)methyl)-3,5-dimethoxyphenyl)-N-(1-(5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)pentyl)piperidin-4-yl)-5-methyl-4-oxo-4,5-dihydrothieno[3,2-c]pyridine-2-carboxamide trifluoroacetic acid (12.8 mg, 26%) as a white solid. ¹H NMR (300 MHz, Methanol-d4) δ 8.25 (s, 1H), 7.87 (s, 1H), 7.67 (t, J=7.8 Hz, 1H), 7.39 (d, J=7.2 Hz, 1H), 7.23 (d, J=8.4 Hz, 1H), 7.06 (s, 2H), 5.16 (dd, J=12.8, 5.4 Hz, 1H), 4.42 (s, 2H), 4.24-4.10 (m, 1H), 4.02 (s, 6H), 3.88 (t, J=6.8 Hz, 2H), 3.75 (s, 3H), 3.69 (s, 2H), 3.25-3.06 (m, 4H), 2.96-2.87 (m, 8H), 2.77-2.60 (m, 1H), 2.31-2.14 (m, 3H), 2.01-1.88 (m, 2H), 1.86-1.62 (m, 4H), 1.51-1.39 (m, 2H). LCMS (ESI) m/z: [M+H]⁺=827.40.

Example 23—Preparation of (3R) and (3S) N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxo thieno[3,2-c]pyridin-2-yl)-1-(5-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]pentanoyl)piperidine-3-carboxamide (Compounds D7 and D8)

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To a stirred solution of N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl)piperidine-3-carboxamide (50.00 mg, 0.103 mmol, 1.00 equiv) and 5-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]pentanoic acid (46.35 mg, 0.124 mmol, 1.20 equiv) in DMF (1 mL) was added DIEA (26.67 mg, 0.206 mmol, 2.00 equiv) dropwise at room temperature. The resulting mixture was stirred for 15 minutes at room temperature. Then HATU (58.84 mg, 0.155 mmol, 1.50 equiv) was added to the reaction mixture. The resulting mixture was stirred for 1 hour at room temperature. The crude products were purified by Prep-HPLC (conditions: SunFire C18 OBD Prep Column, 100 Å, 5 μm, 19 mm×250 mm; mobile phase, Water (0.1% FA) and ACN (7% Phase B up to 21% in 12 minutes); Detector, UV).

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(3R)—N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl)-1-(5-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]pentanoyl)piperidine-3-carboxamide (9.7 mg, 11%) as a white semi-solid. ¹H NMR (400 MHz, Methanol-d4) δ 7.78-7.61 (m, 2H), 7.42 (dd, J=8.0, 2.5 Hz, 1H), 7.34 (d, J=8.5 Hz, 0.5H), 7.23 (d, J=7.2 Hz, 0.5H), 7.18 (s, 0.5H), 7.06 (s, 0.5H), 7.04 (d, J=2.3 Hz, 2H), 5.10-4.92 (m, 1H), 4.41 (d, J=5.1 Hz, 2H), 4.31-4.18 (m, 3H), 4.14 (d, J=12.6 Hz, 0.5H), 4.03 (d, J=2.8 Hz, 6H), 3.89 (d, J=14.1 Hz, 0.5H), 3.73 (d, J=5.0 Hz, 3H), 3.52-3.43 (m, 1H), 3.26-3.14 (m, 1H), 2.92 (d, J=4.2 Hz, 6H), 2.78-2.55 (m, 6H), 2.13-1.99 (m, 2H), 1.97-1.84 (m, 6H), 1.68-1.47 (m, 1H). LCMS (ESI) m/z: [M+H]+=841.32.

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(3S)—N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl)-1-(5-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]pentanoyl)piperidine-3-carboxamide (11.3 mg, 13%) as a white semi-solid. ¹H NMR (300 MHz, Methanol-d4) δ 7.79-7.71 (m, 0.5H), 7.64 (s, 1H), 7.60 (d, J=7.9 Hz, 0.5H), 7.43 (d, J=7.8 Hz, 1H), 7.35 (d, J=8.4 Hz, 1H), 7.19 (s, 0.5H), 7.05 (s, 2H), 7.04 (s, 0.5H), 6.92 (s, 0.5H), 5.12-5.08 (m, 0.5H), 4.96-4.92 (m, 0.5H), 4.42 (d, J=4.5 Hz, 2H), 4.35-4.22 (m, 3H), 4.21-4.14 (m, 0.5H), 4.03 (s, 6H), 3.96-3.88 (m, 0.5H), 3.74 (d, J=5.8 Hz, 3H), 3.43-3.40 (m, 1H), 3.24-3.09 (m, 1H), 2.92 (s, 6H), 2.88-2.49 (m, 6H), 2.16-2.00 (m, 2H), 1.99-1.76 (m, 6H), 1.69-1.42 (m, 1H). LCMS (ESI) m/z: [M+H]+=841.32.

Example 24—Preparation of N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl)-1-(5-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]pentyl)piperidine-3-carboxamide (Compound D9)

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A solution of N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl)piperidine-3-carboxamide (30.00 mg, 0.062 mmol, 1.00 equiv) and 5-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]pentanal (26.62 mg, 0.074 mmol, 1.20 equiv) in DMF (1.5 mL) was stirred for 20 minutes at room temperature. Then NaBH₃CN (7.78 mg, 0.124 mmol, 2.00 equiv) was added to the reaction mixture. The resulting mixture was stirred for 2 hours at room temperature. The crude product was purified by Prep-HPLC (conditions: SunFire 018 OBD Prep Column, 100 Å, 5 μm, 19 mm×250 mm; mobile phase, Water (0.1% FA) and ACN (7% Phase B up to 21% in 12 minutes); Detector, UV). This resulted in N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl)-1-(5-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]pentyl) piperidine-3-carboxamide (5.2 mg, 10%) as a red semi-solid. ¹H NMR (400 MHz, Methanol-d4) δ 8.45 (brs, 0.2H, FA), 7.66 (d, J=5.8 Hz, 1H), 7.62-7.55 (m, 1H), 7.33-7.12 (m, 3H), 7.04 (d, J=2.0 Hz, 2H), 5.14-5.04 (m, 1H), 4.41 (s, 2H), 4.01 (d, J=5.8 Hz, 6H), 3.86 (d, J=7.5 Hz, 2H), 3.74 (s, 3H), 3.31-3.12 (m, 3H), 3.04-2.95 (m, 3H), 2.91 (s, 6H), 2.89-2.82 (m, 2H), 2.73-2.60 (m, 1H), 2.10-1.93 (m, 3H), 1.90-1.73 (m, 4H), 1.66 (t, J=7.1 Hz, 2H), 1.48-1.38 (m, 2H). LCMS (ESI) m/z: [M+H]⁺=827.80.

Example 25—Preparation of 7-(4-((dimethylamino)methyl)-3,5-dimethoxyphenyl)-N-(1-(5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)pentanoyl)piperidin-4-yl)-5-methyl-4-oxo-4,5-dihydrothieno[3,2-c]pyridine-2-carboxamide Formic Acid (Compound D10 Formic Acid)

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Into a stirred mixture of 7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxo-N-(piperidin-4-yl)thieno[3,2-c]pyridine-2-carboxamide (25.0 mg, 0.052 mmol, 1.00 equiv) and 5-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]pentanoic acid (19.3 mg, 0.052 mmol, 1.00 equiv) in DMF (0.50 mL) was added HATU (49.0 mg, 0.129 mmol, 2.50 equiv) at 0° C. under nitrogen atmosphere. After 10 minutes, to the above mixture was added DIEA (33.3 mg, 0.258 mmol, 5.00 equiv) at 0° C. under nitrogen atmosphere and stirred for 10 minutes. Then the mixture was allowed to stir at room temperature for 2 hours. The crude product was purified by Prep-HPLC (conditions: Sunfire C18 OBD Prep Column, 100A, 5 μm, 19 mm*250 mm; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 25 mL/minute; Gradient: 12% B to 35% B in 15 minutes; 254/220 nm; R_T: 11.34 minutes). This resulted in 7-(4-((dimethylamino)methyl)-3,5-dimethoxyphenyl)-N-(1-(5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)pentanoyl)piperidin-4-yl)-5-methyl-4-oxo-4,5-dihydrothieno[3,2-c]pyridine-2-carboxamide; formic acid (7.8 mg, 17%) as a white solid. ¹H NMR (300 MHz, Methanol-d4) δ 8.56 (br s, 0.9H, FA), 8.23 (s, 1H), 7.84 (s, 1H), 7.83-7.76 (m, 1H), 7.47 (d, J=8.8 Hz, 2H), 7.06 (s, 2H), 5.14-5.05 (m, 1H), 4.61-4.52 (m, 1H), 4.36-4.27 (m, 4H), 4.16-4.05 (m, 2H), 4.01 (s, 6H), 3.74 (s, 3H), 2.85 (s, 7H), 2.81-2.73 (m, 3H), 2.69-2.59 (m, 3H), 2.14-2.06 (m, 1H), 2.04-1.87 (m, 6H), 1.66-1.47 (m, 2H). LCMS (ESI) m/z: [M+H]⁺=841.50.

Compound D12 was prepared in a similar manner as described in compound D10.

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N-(7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridin-2-yl)-1-(5-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]pentanoyl)piperidine-4 carboxamide formic acid (8.2 mg, 15%) as a light yellow solid. ¹H NMR (400 MHz, Methanol-d4) δ 8.57 (brs, 0.7H, FA), 7.79 (dd, J=8.6, 7.3 Hz, 1H), 7.64 (s, 1H), 7.45 (dd, J=7.6, 3.3 Hz, 2H), 7.20 (d, J=3.1 Hz, 1H), 7.03 (s, 2H), 5.10-5.04 (m, 1H), 4.62-4.56 (m, 1H), 4.32-4.23 (m, 4H), 4.16-4.08 (m, 1H), 4.00 (s, 6H), 3.74 (s, 3H), 3.23-3.15 (m, 1H), 2.79 (s, 6H), 2.76-2.59 (m, 6H), 2.15-2.07 (m, 1H), 1.99-1.84 (m, 7H), 1.81-1.58 (m, 2H). LCMS (ESI) m/z: [M+H]+=841.35.

Example 26—Preparation of 7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-N-[1-(5-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]pentyl)-3,3-difluoropiperidin-4-yl]-5-methyl-4-oxothieno[3,2-c]pyridine-2-carboxamide Formic Acid (Compound D13 Formic Acid)

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To a solution of N-(3,3-difluoropiperidin-4-yl)-7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridine-2-carboxamide (40.0 mg, 0.077 mmol, 1.00 equiv) and 5-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]pentanal (30.29 mg, 0.085 mmol, 1.1 equiv) in DMF (1 mL) was added NaBH(AcO)₃(32.6 mg, 0.154 mmol, 2 equiv). The resulting solution was stirred at room temperature for 2 hours. Without any additional work-up, the mixture was purified by Prep-HPLC (conditions: SunFire 018 OBD Prep Column, 100 Å, 5 μm, 19 mm×250 mm; Mobile Phase A: Water (0.05% FA), Mobile Phase B: ACN; Flow rate:25 mL/minute; Gradient:13 B to 25 B in 10 minutes; 254 nm) to afford 7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-N-[1-(5-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]pentyl)-3,3-difluoropiperidin-4-yl]-5-methyl-4-oxothieno[3,2-c]pyridine-2-carboxamide formic acid (20.9 mg, 29.78%) as a white solid. ¹H NMR (400 MHz, DMSO-d6) δ 11.11 (s, 1H), 8.87 (d, J=8.8 Hz, 1H), 8.59 (s, 1H), 8.15 (s, 0.3H, FA), 7.97 (s, 1H), 7.82 (t, J=7.9 Hz, 1H), 7.54 (d, J=8.6 Hz, 1H), 7.45 (d, J=7.3 Hz, 1H), 6.96 (s, 2H), 5.09 (dd, J=12.9, 5.4 Hz, 1H), 4.38-4.20 (m, 3H), 3.88 (s, 6H), 3.79 (s, 2H), 3.63 (s, 3H), 3.15 (s, 1H), 2.96-2.83 (m, 2H), 2.70-2.56 (m, 3H), 2.41 (s, 8H), 2.19 (t, J=11.4 Hz, 1H), 2.08-2.00 (m, 1H), 1.92-1.73 (m, 4H), 1.58-1.43 (m, 4H). LCMS (ESI) m/z: [M+H]+=863.35.

Example 27—Preparation of 7-(4-[[([[2-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4 yl]oxy]ethoxy)ethyl] carbamoyl]methyl)(methyl)amino]methyl]-3,5-dimethoxyphenyl)-5-methyl-N-(1-methylpiperidin-4-yl)-4-oxothieno[3,2-c]pyridine-2-carboxamide Formic Acid (Compound D14 Formic Acid)

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To a stirred mixture of [[(2,6-dimethoxy-4-[5-methyl-2-[(1-methylpiperidin-4-yl)carbamoyl]-4-oxothieno[3,2-c]pyridin-7-yl]phenyl)methyl](methyl)amino]acetic acid (50.00 mg, 0.076 mmol, 1.00 equiv) and 4-[2-(2-aminoethoxy)ethoxy]-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione trifluoroacetic acid (36.20 mg, 0.076 mmol, 1.00 equiv) in DMF (1.00 mL) was added HATU (72.38 mg, 0.190 mmol, 2.50 equiv) at 0° C. under atmosphere. After 10 minutes, DIEA (49.20 mg, 0.381 mmol, 5.00 equiv) was added at 0° C. under nitrogen atmosphere, and the reaction mixture was then stirred for 10 minutes. Then the mixture was allowed to stir at room temperature for 3 hours. The crude product (0.1% TA) was concentrated, purified by Prep-TLC (DCM/MeOH=10:1) to afford crude product. Then the crude product was purified by Prep-HPLC (conditions: Sunfire C18 OBD Prep Column, 100 Å, 5 μm, 19 mm*250 mm; Mobile Phase A: Water (0.05% FA), Mobile Phase B: ACN; Flow rate: 25 mL/minute; Gradient: 12 B to 35 B in 12 minutes; 254 nm; R_T: 11.68 minutes). This resulted in 7-(4-[[([[2-(2-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxy]ethoxy)ethyl]carbamoyl]methyl)(methyl)amino]methyl]-3,5-dimethoxyphenyl)-5-methyl-N-(1-methylpiperidin-4-yl)-4-oxothieno [3,2-c]pyridine-2-carboxamide formic acid (3.0 mg, 7.96%) as a white solid. ¹H NMR (400 MHz, Methanol-d4) δ 8.45 (s, 1.0H, FA), 8.21 (s, 1H), 7.78 (s, 1H), 7.64 (dd, J=8.6, 7.2 Hz, 1H), 7.33 (dd, J=7.8, 2.1 Hz, 2H), 6.88 (s, 2H), 5.07 (dd, J=12.7, 5.5 Hz, 1H), 4.32-4.26 (m, 2H), 4.17-4.08 (m, 1H), 4.02 (s, 2H), 3.92 (s, 6H), 3.90-3.86 (m, 2H), 3.73 (s, 3H), 3.69 (t, J=5.2 Hz, 2H), 3.54-3.43 (m, 6H), 3.09 (t, J=12.2 Hz, 2H), 2.91-2.80 (m, 4H), 2.78-2.61 (m, 2H), 2.56 (s, 3H), 2.27-2.18 (m, 2H), 2.14-2.06 (m, 1H), 2.02-1.89 (m, 2H). LCMS (ESI) m/z: [M+H]⁺=886.20.

Example 28—Preparation of 7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-N-[1-(5-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]pentyl)-3,3-difluoropiperidin-4-yl]-5-methyl-4-oxothieno[3,2-c]pyridine-2-carboxamide Formic Acid (Compound D15 Formic Acid)

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To a solution of N-(3,3-difluoropiperidin-4-yl)-7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-5-methyl-4-oxothieno[3,2-c]pyridine-2-carboxamide (40.0 mg, 0.077 mmol, 1.00 equiv) and 5-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]pentanal (30.3 mg, 0.085 mmol, 1.1 equiv) in DMF (1 mL) was added NaBH(AcO)₃(32.6 mg, 0.154 mmol, 2 equiv). The resulting solution was stirred at room temperature for 2 hours. Without any additional work-up, the mixture was purified by Prep-HPLC (conditions: SunFire 018 OBD Prep Column, 100 Å, 5 μm, 19 mm×250 mm; Mobile Phase A: Water (0.05% FA), Mobile Phase B:ACN; Flow rate:25 mL/minute; Gradient:15 B to 25 B in 12 minutes; 254 nm) to afford 7-[4-[(dimethylamino)methyl]-3,5-dimethoxyphenyl]-N-[1-(5-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]oxy]pentyl)-3,3-difluoropiperidin-4-yl]-5-methyl-4-oxothieno[3,2-c]pyridine-2-carboxamide formic acid (14 mg, 19.81%) as a white solid. ¹H NMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 8.88 (d, J=8.8 Hz, 1H), 8.60 (s, 1H), 8.15 (s, 0.3H, FA), 7.98 (s, 1H), 7.84 (d, J=8.3 Hz, 1H), 7.44 (d, J=2.3 Hz, 1H), 7.36 (dd, J=8.3, 2.3 Hz, 1H), 6.97 (s, 2H), 6.53 (s, 0.3H, FA salt), 5.13 (dd, J=12.9, 5.3 Hz, 1H), 4.35 (s, 1H), 4.20 (t, J=6.5 Hz, 2H), 3.88 (s, 6H), 3.82 (s, 2H), 3.63 (s, 3H), 3.15 (s, 1H), 2.95-2.84 (m, 2H), 2.65-2.55 (m, 3H), 2.44 (s, 8H), 2.18 (t, J=11.3 Hz, 1H), 2.10-2.01 (m, 1H), 1.91-1.74 (m, 4H), 1.57-1.41 (m, 4H). LCMS (ESI) m/z: [M+H]+=863.35.

Example 29—Preparation of Compounds D16-D90

In analogy to the procedures described in the examples above, compounds D16-D90 were prepared using the appropriate starting materials

Compound

No.
LCMS

¹H NMR

D16
LCMS (ESI) m/z:

¹H NMR (400 MHz, DMSO-d6) δ 11.11 (s, 1H), 8.87 (d, J = 8.8 Hz,

[M + H]+ = 863.35
1H), 8.59 (s, 1H), 8.15 (s, 0.3H, FA), 7.97 (s, 1H), 7.82 (t, J = 7.9

Hz, 1H), 7.54 (d, J = 8.6 Hz, 1H), 7.45 (d, J = 7.3 Hz, 1H), 6.96 (s,

2H), 5.09 (dd, J = 12.9, 5.4 Hz, 1H), 4.38-4.20 (m, 3H), 3.88 (s,

6H), 3.79 (s, 2H), 3.63 (s, 3H), 3.15 (s, 1H), 2.96-2.83 (m, 2H),

2.70-2.56 (m, 3H), 2.41 (s, 8H), 2.19 (t, J = 11.4 Hz, 1H), 2.08-

2.00 (m, 1H), 1.92-1.73 (m, 4H), 1.58-1.43 (m, 4H).

D17
879.15

¹H NMR (300 MHz, DMSO-d6) δ 10.96 (s, 1H), 8.89 (d, J = 8.8 Hz,

1H), 8.60 (s, 1H), 8.17 (s, 2H, FA), 8.01 (s, 1H), 7.53 (d, J = 8.8

Hz, 1H), 7.34 (d, J = 1.6 Hz, 1H), 7.25 (d, J = 1.7 Hz, 1H), 7.07 (d,

J = 7.6 Hz, 2H), 5.15-4.95 (m, 1H), 4.47-4.14 (m, 3H), 3.89 (s,

3H), 3.62 (s, 3H), 3.59-3.52 (m, 2H), 3.33-3.14 (m, 4H), 3.03-

2.79 (m, 3H), 2.69-2.54 (m, 8H), 2.46-2.26 (m, 3H), 2.25-2.16

(m, 7H), 2.05-1.90 (m, 1H), 1.90-1.69 (m, 2H).

D18
929.25

¹H NMR (300 MHz, DMSO-d6) δ 10.95 (s, 1H), 8.95 (d, J = 9.0 Hz,

1H), 8.62 (d, J = 13.0 Hz, 1H), 8.14 (s, 0.47H, FA), 8.00 (s, 1H),

7.50 (d, J = 8.2 Hz, 1H), 7.00 (s, 2H), 6.56-6.45 (m, 2H), 5.04

(dd, J = 13.2, 5.1 Hz, 1H), 4.62 (s, 2H), 4.31 (d, J = 16.9 Hz, 1H),

4.18 (d, J = 17.0 Hz, 2H), 4.00 (s, 2H), 3.90 (s, 6H), 3.65 (d, J =

9.8 Hz, 7H), 3.22 (dd, J = 15.8, 3.3 Hz, 2H), 3.01-2.84 (m, 3H),

2.64-2.53 (m, 6H), 2.44-2.31 (m, 5H), 2.02-1.88 (m, 3H), 1.88-

1.66 (m, 4H).

D19
915.25

¹H NMR (300 MHz, DMSO-d6) δ 10.96 (s, 1H), 8.98 (s, 1H, TFA

salt), 8.92 (d, J = 8.8 Hz, 1H), 8.61 (s, 1H), 8.02 (s, 1H), 7.53 (d,

J = 8.2 Hz, 1H), 7.03 (s, 2H), 6.56-6.48 (m, 2H), 5.05 (dd, J = 13.1,

5.0 Hz, 1H), 4.50-4.10 (m, 7H), 3.93 (s, 6H), 3.74 (s, 4H), 3.64

(s, 3H), 3.04-2.84 (m, 4H), 2.81-2.76 (m, 8H), 2.69-2.55 (m,

5H), 2.40-2.30 (m, 2H), 2.12-1.94 (m, 5H), 1.91-1.75 (m,

2H).

D20
752.2

¹H NMR (400 MHz, DMSO-d6) δ 10.97 (s, 1H), 9.63 (s, 1H), 8.23

(s, 2H, FA), 8.14 (s, 1H), 7.37 (d, J = 8.1 Hz, 1H), 7.14 (s, 2H),

6.72-6.64 (m, 2H), 5.13-5.04 (m, 1H), 4.31 (d, J = 16.6 Hz, 1H),

4.18 (d, J = 16.6 Hz, 1H), 3.86 (s, 6H), 3.81 (s, 2H), 3.68 (s, 3H),

3.56 (s, 6H), 3.17 (s, 2H), 2.91 (m, J = 17.7, 13.4, 5.4 Hz, 1H),

2.59 (d, J = 17.0 Hz, 2H), 2.45 (d, J = 6.9 Hz, 2H), 2.40-2.22 (m,

5H), 2.04-1.93 (m, 1H), 1.72 (t, J = 5.4 Hz, 4H).

D21
734.34

¹H NMR (300 MHz, DMSO-d6) δ 10.99 (s, 1H), 9.87 (d, J = 53.7

Hz, 2H, TFA), 8.26 (d, J = 2.0 Hz, 1H), 7.90 (s, 1H), 7.41 (d, J =

8.8 Hz, 1H), 7.14 (d, J = 2.1 Hz, 1H), 6.91 (d, J = 2.2 Hz, 2H), 6.70

(dd, J = 4.8, 2.4 Hz, 2H), 5.07 (dd, J = 13.3, 5.1 Hz, 1H), 4.50-

4.28 (m, 3H), 4.20 (d, J = 17.0 Hz, 2H), 3.95 (s, 6H), 3.73 (s, 2H),

3.65 (s, 5H), 2.96 (s, 3H), 2.78 (d, J = 4.7 Hz, 2H), 2.63 (s, 2H),

2.43-2.31 (m, 1H), 2.12 (d, J = 13.6 Hz, 2H), 2.03-1.79 (m, 3H).

D22
818.15

¹H NMR (400 MHz, DMSO-d6) δ 10.97 (s, 1H), 9.51 (s, 1H, TFA),

9.05 (s, 1H, TFA), 8.61 (s, 1H), 8.03 (s, 1H), 7.60 (d, J = 8.4 Hz,

1H), 7.19-7.10 (m, 2H), 7.06 (s, 2H), 5.07 (dd, J = 13.3, 5.1 Hz,

1H), 4.64 (s, 1H), 4.44-4.30 (m, 3H), 4.24 (d, J = 17.0 Hz, 1H),

3.99 (d, J = 12.7 Hz, 2H), 3.96 (s, 6H), 3.76 (s, 1H), 3.65 (s,

3H), 3.50 (m, 1H), 3.40 (m, 1H), 3.25 (m, 1H), 3.21 (m, 2H), 3.00-

2.70 (m, 5H), 2.69-2.55 (m, 2H), 2.45-2.29 (m, 2H), 2.09 (s,

2H), 2.03-1.93 (m, 1H).

D23
854.35

¹H NMR (400 MHz, Methanol-d4) δ 8.36 (s, 1H), 8.30 (s, 0.3H,

FA), 7.85 (s, 1H), 7.76 (d, J = 8.5 Hz, 1H), 7.46 (d, J = 2.3 Hz, 1H),

7.33 (dd, J = 8.6, 2.3 Hz, 1H), 7.05 (s, 2H), 5.11 (dd, J = 12.5, 5.5

Hz, 1H), 4.50-4.37 (m, 1H), 4.31 (s, 2H), 4.02 (s, 6H), 3.75 (s,

3H), 3.69 (s, 4H), 3.40-3.35 (m, 2H), 3.31-3.21 (m, 2H), 3.19

(t, J = 9.9 Hz, 1H), 2.97 (d, J = 12.0 Hz, 1H), 2.89 (ddd, J = 18.3,

14.2, 5.1 Hz, 1H), 2.78 (d, J = 3.2 Hz, 1H), 2.73 (d, J = 9.9 Hz,

1H), 2.47 (dd, J = 28.7, 12.4 Hz, 1H), 2.40 (s, 3H), 2.31 (t, J = 11.7

Hz, 1H), 2.19-2.04 (m, 1H), 2.01 (dd, J = 11.9, 4.4 Hz, 1H), 1.96

(d, J = 7.4 Hz, 1H).

D24
LCMS (ESI) m/z:

¹H NMR (300 MHz, Methanol-d4) δ 8.37 (d, J = 1.8 Hz, 1H), 7.89

[M + H]+ = 846.25
(s, 1H), 7.49 (d, J = 8.2 Hz, 1H), 7.37 (d, J = 11.8 Hz, 2H), 7.08 (s,

2H), 5.16 (dd, J = 13.3, 5.0 Hz, 1H), 4.74 (m, 1H) 4.43 (d, J = 6.3

Hz, 4H), 4.10 (d, J = 12.5 Hz, 1H), 4.02 (s, 6H), 3.95 (d, J = 12.6

Hz, 2H), 3.87 (s, 1H), 3.76 (s, 3H), 3.53 (d, 2H), 3.22 (s, 1H), 2.92

(s, 8H), 2.89-2.74 (m, 2H), 2.60-2.42 (m, 1H), 2.21 (s, 5H),

1.90 (t, J = 11.8 Hz, 2H).

D25
LCMS (ESI) m/z:

¹H NMR (300 MHz, Methanol-d4) δ 8.36 (s, 1H), 7.88 (s, 1H), 7.67

[M + H]⁺ = 846.25.
(d, J = 9.2 Hz, 1H), 7.13-7.03 (t, 4H), 5.12 (dd, J = 13.2, 5.1 Hz,

1H), 4.75 (s, 1H), 4.52-4.33 (m, 4H), 4.12 (d, J = 12.9 Hz, 2H),

4.02 (s, 6H), 3.93 (s, 1H), 3.75 (s, 3H), 3.65-3.46 (m, 3H), 3.25

(s, 1H), 2.99 (d, J = 12.6 Hz, 2H), 2.92-2.73 (m, 7H), 2.57-2.40

(m, 1H), 2.27 (s, 1H), 2.21 (s, 5H), 1.90 (q, J = 12.2, 11.6 Hz, 2H).

D26
LCMS (ESI) m/z:

¹H NMR (300 MHz, DMSO-d6, D₂O) δ 8.45 (s, 1H), 7.90 (s, 1H),

[M + H]⁺ = 943.45.
7.75 (d, J = 8.5 Hz, 1H), 7.40 (d, J = 2.0 Hz, 1H), 7.31 (dd, J = 8.6,

2.3 Hz, 1H), 6.99-6.95 (m, 2H), 5.02 (dd, J = 12.8, 5.6 Hz, 1H),

4.76-4.58 (m, 1H), 4.35-4.12 (m, 4H), 3.96-3.83 (m, 7H), 3.72-

3.48 (m, 7H), 3.39 (d, J = 11.5 Hz, 2H), 3.31-3.07 (m, 7H), 2.97

(t, J = 12.3 Hz, 2H), 2.86 (s, 3H), 2.83-2.72 (m, 1H), 2.69-2.55

(m, 2H), 2.19-2.00 (m, 3H), 1.87-1.56 (m, 5H), 1.49-1.32 (m,

2H).

D27
LCMS (ESI) m/z:

¹H NMR (300 MHz, DMSO-d6, D₂O) δ 8.45 (s, 1H), 7.90 (s, 1H),

[M + H]⁺ = 935.55.
7.75 (d, J = 8.5 Hz, 1H), 7.40 (d, J = 2.0 Hz, 1H), 7.31 (dd, J = 8.6,

2.3 Hz, 1H), 6.99-6.95 (m, 2H), 5.02 (dd, J = 12.8, 5.6 Hz, 1H),

4.76-4.58 (m, 1H), 4.35-4.12 (m, 4H), 3.96-3.83 (m, 7H), 3.72-

3.48 (m, 7H), 3.39 (d, J = 11.5 Hz, 2H), 3.31-3.07 (m, 7H), 2.97

(t, J = 12.3 Hz, 2H), 2.86 (s, 3H), 2.83-2.72 (m, 1H), 2.69-2.55

(m, 2H), 2.19-2.00 (m, 3H), 1.87-1.56 (m, 5H), 1.49-1.32 (m,

2H).

D28
LCMS (ESI) m/z:

¹H NMR (300 MHz, Methanol-d4) δ 8.58 (s, 1H, FA), 8.35 (s, 1H),

[M + H]⁺ = 875.50.
7.85-7.78 (m, 2H), 7.42 (d, J = 2.2 Hz, 1H), 7.33 (dd, J = 8.3, 2.3

Hz, 1H), 7.02 (s, 2H), 5.12 (dd, J = 12.2, 5.4 Hz, 1H), 4.52-4.39

(m, 1H), 4.36 (s, 2H), 4.20 (t, J = 6.3 Hz, 2H), 4.06 (t, J = 8.0 Hz,

4H), 4.00 (s, 6H), 3.74 (s, 3H), 3.26-3.18 (m, 1H), 3.07-2.97

(m, 1H), 2.89-2.69 (m, 3H), 2.59-2.36 (m, 5H), 2.34-2.23 (m,

1H), 2.20-2.07 (m, 1H), 2.02-1.85 (m, 4H), 1.69-1.54 (m, 4H).

D29
LCMS (ESI) m/z:

¹H NMR (300 MHz, Methanol-d4) δ 8.52 (s, 2H, FA), 8.22 (s, 1H),

[M + H]⁺ = 853.45.
7.86-7.76 (m, 2H), 7.46 (dd, J = 8.3, 2.4 Hz, 2H), 7.05 (s, 2H),

5.09 (dd, J = 12.5, 5.2 Hz, 1H), 4.55 (d, J = 13.7 Hz, 1H), 4.48 (s,

2H), 4.34-4.26 (m, 2H), 4.20 (t, J = 8.2 Hz, 4H), 4.14-4.05 (m,

2H), 4.02 (s, 6H), 3.74 (s, 3H), 3.29-3.18 (m, 1H), 2.86-2.60

(m, 6H), 2.58-2.46 (m, 2H), 2.14-1.87 (m, 7H), 1.69-1.44 (m,

2H).

D30
LCMS (ESI) m/z:

¹H NMR (400 MHz, DMSO-d6 with a drop of D2O) δ 9.07 (d, J =

[M + H]⁺ = 922.35
8.5 Hz, 1H, FA salt), 8.49 (s, 1H), 7.91 (s, 1H), 7.76 (dd, J = 8.5,

7.3 Hz, 1H), 7.42 (dd, J = 11.9, 7.9 Hz, 2H), 6.92 (s, 2H), 5.01 (dd,

J = 12.8, 5.5 Hz, 1H), 4.53 (br s, 1H), 4.28 (s, 4H), 3.85-3.31 (m,

6H), 3.78 (s, 4H), 3.60 (s, 3H), 3.55 (d, J = 5.6 Hz, 2H), 3.35-

3.21 (m, 3H), 2.87-2.55 (m, 10H), 2.48-2.40 (m, 2H), 2.09-

1.93 (m, 3H).

D31
LCMS (ESI) m/z:

¹H NMR (400 MHz, DMSO-d6) δ 11.10 (s, 1H), 8.27 (s, 1H), 8.14

[M + H]⁺ = 925.15.
(s, 0.4H, FA), 7.98 (dd, J = 5.3, 3.5 Hz, 2H), 7.96-7.86 (m, 2H),

7.79 (d, J = 7.0 Hz, 2H), 7.56 (dd, J = 8.6, 7.1 Hz, 1H), 7.03 (dd,

J = 13.0, 7.8 Hz, 2H), 6.86 (s, 1H), 6.48 (t, J = 5.9 Hz, 1H), 5.05 (dd,

J = 12.7, 5.4 Hz, 1H), 3.85 (s, 2H), 3.71 (d, J = 16.7 Hz, 1H), 3.60

(s, 3H), 3.52-3.37 (m, 2H), 3.23 (q, J = 6.5 Hz, 3H), 2.99 (d, J =

6.6 Hz, 2H), 2.88 (ddd, J = 16.7, 13.7, 5.2 Hz, 1H), 2.59 (d, J =

18.0 Hz, 2H), 2.49-2.41 (m, 1H), 2.22 (s, 1H), 2.06-1.94 (m,

1H), 1.66-1.45 (m, 2H), 1.42-1.00 (m, 11H).

D32
LCMS (ESI) m/z:

¹H NMR (400 MHz, Methanol-d4) δ 8.45 (s, 1.0H, FA), 8.21 (s,

[M + H]⁺ = 886.20
1H), 7.78 (s, 1H), 7.64 (dd, J = 8.6, 7.2 Hz, 1H), 7.33 (dd, J = 7.8,

2.1 Hz, 2H), 6.88 (s, 2H), 5.07 (dd, J = 12.7, 5.5 Hz, 1H), 4.32-

4.26 (m, 2H), 4.17-4.08 (m, 1H), 4.02 (s, 2H), 3.92 (s, 6H), 3.90-

3.86 (m, 2H), 3.73 (s, 3H), 3.69 (t, J = 5.2 Hz, 2H), 3.54-3.43

(m, 6H), 3.09 (t, J = 12.2 Hz, 2H), 2.91-2.80 (m, 4H), 2.78-2.61

(m, 2H), 2.56 (s, 3H), 2.27-2.18 (m, 2H), 2.14-2.06 (m, 1H),

2.02-1.89 (m, 2H).

D33
LCMS (ESI) m/z:

[M + H]+ = 921.65

D34
LCMS (ESI) m/z:

[M + H]+ = 921.45

D35
LCMS (ESI) m/z:

[M + H]+ = 935.6

D36
LCMS (ESI) m/z:

[M + H]+ = 907.4

D37
LCMS (ESI) m/z:

[M + H]+ = 893.45

D38
LCMS (ESI) m/z:

[M + H]+ = 879.25

D39
LCMS (ESI) m/z:

[M + H]+ = 893.5

D40
LCMS (ESI) m/z:

[M + H]+ = 921.75

D41
LCMS (ESI) m/z:

[M + H]+ = 893.15

D42
LCMS (ESI) m/z:

[M + H]+ = 907.65

D43
LCMS (ESI) m/z:

¹H NMR (400 MHz, DMSO-d6) δ 11.07 (s, 1H), 8.82-8.73 (m,

[M + H]+ = 893.3
1H), 8.36 (d, J = 4.1 Hz, 1H), 8.22 (s, 2H, FA), 7.92 (d, J = 4.3 Hz,

1H), 7.65 (dd, J = 8.5, 6.6 Hz, 1H), 7.32 (d, J = 2.3 Hz, 1H), 7.26-

7.18 (m, 1H), 6.91 (s, 2H), 5.11-5.00 (m, 1H), 3.85 (d, J = 2.3 Hz,

6H), 3.61 (d, J = 3.9 Hz, 6H), 3.45-3.38 (m, 4H), 3.25 (t, J = 5.8

Hz, 1H), 3.19 (t, J = 6.5 Hz, 1H), 2.93-2.83 (m, 1H), 2.72-2.66

(m, 1H), 2.65-2.60 (m, 2H), 2.59-2.53 (m, 7H), 2.48-2.44 (m,

3H), 2.26 (d, J = 2.3 Hz, 8H), 2.05-1.94 (m, 3H), 1.55 (dd, J =

12.5, 6.1 Hz, 1H), 1.46-1.36 (m, 1H), 1.08-0.97 (m, 1H).

D44
LCMS (ESI) m/z:

[M + H]+ = 893.3

D45
929.45

D46
943.45

¹H NMR (300 MHz, DMSO-d6) δ 11.07 (s, 1H), 8.92 (d, J = 8.6 Hz,

1H), 8.59 (d, J = 8.7 Hz, 1H), 8.22 (s, 1H, FA), 7.96 (s, 1H), 7.66

(d, J = 8.4 Hz, 1H), 6.92 (s, 2H), 6.81 (s, 1H), 6.73-6.63 (m, 1H),

5.06 (dd, J = 12.6, 5.4 Hz, 1H), 4.75-4.07 (m, 4H), 4.04-3.93

(m, 4H), 3.85 (s, 6H), 3.63 (s, 4H), 3.54-3.51 (m, 4H), 2.93-

2.55 (m, 8H), 2.20 (s, 7H), 2.12-2.00 (m, 3H), 1.93-1.82 (m,

2H).

D47
LCMS (ESI) m/z:

[M + H]+ = 461.45

D48
LCMS (ESI) m/z:

[M + H]+ = 951.4

D49
LCMS (ESI) m/z:

¹H NMR (300 MHz, DMSO-d6) δ 11.09 (s, 1H), 8.97 (t, J = 5.8 Hz,

[M + H]+ = 937.4
1H), 8.40 (s, 1H), 8.16 (s, 1H FA), 7.99 (s, 1H), 7.69 (d, J = 8.5 Hz,

1H), 7.35 (d, J = 2.2 Hz, 1H), 7.27 (dd, J = 8.6, 2.2 Hz, 1H), 6.99

(s, 2H), 5.08 (dd, J = 12.7, 5.4 Hz, 1H), 4.09 (t, J = 6.5 Hz, 1H),

4.00 (s, 2H), 3.90 (s, 6H), 3.62 (s, 3H), 3.60-3.49 (m, 4H), 3.46-

3.42 (m, 6H), 3.02-2.60 (m, 9H), 2.59-2.55 (m, 7H), 2.37 (t, J =

7.0 Hz, 2H), 2.07-1.87 (m, 3H), 1.78-1.68 (m, 2H), 1.67-1.57

(m, 4H), 1.49-1.39 (m, 1H).

D50
LCMS (ESI) m/z:

[M + H]+ = 923.4

D51
LCMS (ESI) m/z:

[M + H]+ = 907.4

D52
LCMS (ESI) m/z:

[M + H]+ = 893.4

D53
LCMS (ESI) m/z:

¹H NMR (300 MHz, DMSO-d6) δ 11.09 (s, 1H), 8.80 (t, J = 5.7 Hz,

[M + H]⁺ = 879.
1H), 8.36 (s, 1H), 7.92 (s, 1H), 7.66 (d, J = 8.5 Hz, 1H), 7.32 (d,

J = 2.2 Hz, 1H), 7.23 (dd, J = 8.7, 2.3 Hz, 1H), 6.90 (s, 2H), 5.07

(dd, J = 12.8, 5.4 Hz, 1H), 3.58 (d, J = 12.5 Hz, 5H), 3.37 (d, J =

13.7 Hz, 8H), 3.24 (t, J = 6.2 Hz, 2H), 2.89 (td, J = 17.8, 15.7, 5.3

Hz, 1H), 2.69 (s, 3H), 2.55 (s, 3H), 2.34 (d, J = 6.2 Hz, 3H), 2.22

(s, 6H), 2.17 (d, J = 8.5 Hz, 1H), 2.01 (s, 1H), 1.89 (dd, J = 12.1,

6.6 Hz, 2H).

D54
LCMS (ESI) m/z:

[M + H]+ = 921.25

D55
LCMS (ESI) m/z:

[M + H]+ = 907.35

D56
LCMS (ESI) m/z:

[M + H]+ = 454.5

D57
LCMS (ESI) m/z:

[M + H]+ = 893.35

D58
LCMS (ESI) m/z:

[M + H]+ = 907.35

D59
LCMS (ESI) m/z:

¹H NMR (400 MHz, Methanol-d4) δ 8.56 (s, 1H, FA), 7.83-7.74

[M + H]⁺ = 863.50.
(m, 1H), 7.67 (s, 1H), 7.45 (d, J = 7.8 Hz, 2H), 7.25 (d, J = 1.9 Hz,

1H), 7.06 (s, 2H), 5.10 (dd, J = 12.2, 5.4 Hz, 1H), 4.36 (s, 2H),

4.27 (t, J = 6.1 Hz, 2H), 4.02 (s, 6H), 3.74 (s, 3H), 3.24-3.17 (m,

1H), 3.03-2.93 (m, 2H), 2.88 (s, 6H), 2.83-2.67 (m, 3H), 2.57-

2.42 (m, 3H), 2.27 (t, J = 11.1 Hz, 1H), 2.19-2.09 (m, 2H), 1.99-

1.88 (m, 3H), 1.71-1.57 (m, 4H).

D60

D61
LCMS (ESI) m/z:

[M + H]+ = 469.3

D62
LCMS (ESI) m/z:

[M + H]+ = 909.2

D63
LCMS (ESI) m/z:

¹H NMR (400 MHz, DMSO-d6) δ 11.10 (d, J = 3.2 Hz, 1H), 8.86

[M + H]+ = 893.25
(dd, J = 26.1, 5.8 Hz, 1H), 8.55 (d, J = 38.1 Hz, 1H), 8.15 (s, 1H,

FA), 8.04-7.87 (m, 1H), 7.66 (dd, J = 25.8, 8.5 Hz, 1H), 7.38-

7.24 (m, 1H), 7.20-7.07 (m, 1H), 7.03-6.90 (m, 2H), 5.08 (dd,

J = 12.7, 6.4 Hz, 1H), 4.37-4.25 (m, 1H), 4.04 (s, 2H), 3.89 (d, J =

15.0 Hz, 6H), 3.77-3.53 (m, 7H), 3.45-3.41 (m, 3H), 3.24 (s,

3H), 3.16-3.04 (m, 2H), 3.03-2.73 (m, 4H), 2.64-2.55 (m,

10H), 2.41-2.36 (m, 2H), 2.05-1.85 (m, 2H).

D64
LCMS (ESI) m/z:

[M + H]+ = 865.25

D65
LCMS (ESI) m/z:

[M + H]+ = 879.35

D66
LCMS (ESI) m/z:

[M + H]+ = 879.3

D67
921.5

D68
LCMS (ESI) m/z:

¹H NMR (300 MHz, DMSO-d6) δ 11.08 (s, 1H), 8.80 (dd, J = 11.0,

[M + H]+ = 893.5
5.8 Hz, 1H), 8.54 (s, 1H), 8.14 (s, 1H FA), 8.01-7.92 (m, 1H),

7.65 (dd, J = 18.7, 8.5 Hz, 1H), 7.37-7.15 (m, 2H), 6.99 (d, J =

11.3 Hz, 2H), 5.11-5.01 (m, 1H), 4.48-4.20 (m, 2H), 4.13 (s,

2H), 3.92 (s, 6H), 3.62 (d, J = 9.3 Hz, 3H), 3.48-3.36 (m, 10H),

2.98-2.72 (m, 4H), 2.70-2.59 (m, 9H), 2.42-2.34 (m, 1H), 2.14-

1.92 (m, 2H), 1.78-1.56 (m, 3H).

D69
LCMS (ESI) m/z:

[M + H]+ = 879.4

D70
LCMS (ESI) m/z:

[M + H]+ = 865.25

D71
LCMS (ESI) m/z:

¹H NMR (300 MHz, Methanol-d4) δ 7.81 (d, J = 8.3 Hz, 1H), 7.46

[M + H]⁺ = 813.25.
(s, 1H), 7.42 (d, J = 2.2 Hz, 1H), 7.33 (dd, J = 8.4, 2.3 Hz, 1H),

6.95 (s, 2H), 6.41 (s, 1H), 5.11 (dd, J = 12.4, 5.4 Hz, 1H), 4.37 (d,

J = 13.5 Hz, 1H), 4.22 (t, J = 5.8 Hz, 2H), 4.06 (s, 2H), 3.99-3.92

(m, 7H), 3.71 (s, 3H), 3.59-3.51 (m, 1H), 3.06-2.93 (m, 1H),

2.91-2.68 (m, 4H), 2.66-2.51 (m, 8H), 2.23-2.05 (m, 3H), 1.96-

1.82 (m, 4H), 1.54-1.35 (m, 2H).

D72
LCMS (ESI) m/z:

¹H NMR (300 MHz, DMSO-d6) δ 11.09 (s, 1H), 9.04-8.96 (m,

[M + H]+ = 907.45
1H), 8.39 (s, 1H), 8.29 (s, 2H, FA), 7.94 (s, 1H), 7.68 (dd, J = 8.5,

3.4 Hz, 1H), 7.35 (d, J = 2.7 Hz, 1H), 7.27 (d, J = 7.3 Hz, 1H), 6.91-

6.86 (m, 2H), 5.07 (dd, J = 12.7, 5.3 Hz, 1H), 4.44-4.34 (m,

1H), 3.84 (s, 6H), 3.62 (s, 4H), 3.53-3.48 (m, 10H), 2.93-2.82

(m, 2H), 2.63-2.55 (m, 6H), 2.33-2.23 (m, 3H), 2.21-2.07 (m,

9H), 2.00-1.80 (m, 3H).

D73
LCMS (ESI) m/z:

¹H NMR (300 MHz, DMSO-d6) δ 11.09 (s, 1H), 9.01-8.92 (m,

[M + H]+ = 893.3
1H), 8.39 (s, 1H), 8.23 (s, 3H, FA), 7.95 (s, 1H), 7.68 (d, J = 8.5

Hz, 1H), 7.35 (d, J = 2.2 Hz, 1H), 7.27 (d, J = 8.8 Hz, 1H), 6.91 (s,

2H), 5.08 (dd, J = 12.7, 5.3 Hz, 1H), 4.33-4.24 (m, 1H), 3.85 (s,

6H), 3.62 (s, 5H), 3.44 (s, 5H), 2.97-2.81 (m, 2H), 2.77-2.69

(m, 3H), 2.64-2.53 (m, 5H), 2.39-2.33 (m, 3H), 2.27 (s, 6H),

2.17-2.09 (m, 2H), 2.04-1.84 (m, 4H), 1.68 (s, 2H).

D74
LCMS (ESI) m/z:

¹H NMR (300 MHz, DMSO-d6) δ 11.09 (s, 1H), 9.00-8.93 (m,

[M + H]+ = 893.3
1H), 8.37 (s, 1H), 8.20 (s, 1H, FA), 7.95 (s, 1H), 7.68 (d, J = 8.5

Hz, 1H), 7.35 (d, J = 2.2 Hz, 1H), 7.30-7.23 (m, 1H), 6.91 (s, 2H),

5.08 (dd, J = 12.8, 5.3 Hz, 1H), 4.31-4.19 (m, 2H), 4.12 (s, 1H),

3.92 (s, 1H), 3.87-3.80 (m, 7H), 3.63-3.56 (m, 6H), 3.45-3.40

(m, 8H), 2.91-2.83 (m, 1H), 2.64-2.55 (m, 5H), 2.32-2.20 (m,

10H), 2.08-1.96 (m, 1H).

D75
LCMS (ESI) m/z:

[M + H]+ = 879.45

D76
LCMS (ESI) m/z:

[M + H]+ = 907.35

D77
LCMS (ESI) m/z:

¹H NMR (300 MHz, Methanol-d4) δ 8.24 (s, 1H), 7.87 (s, 1H), 7.80

[M + H]⁺ = 867.50.
(d, J = 8.4 Hz, 1H), 7.52 (d, J = 2.3 Hz, 1H), 7.39 (dd, J = 8.6, 2.3

Hz, 1H), 7.07 (s, 2H), 5.12 (dd, J = 12.3, 5.4 Hz, 1H), 4.60 (d, J =

14.2 Hz, 1H), 4.42 (s, 2H), 4.40-4.29 (m, 2H), 4.20-4.12 (m,

1H), 4.02 (s, 7H), 3.87-3.78 (m, 2H), 3.75 (s, 4H), 3.66-3.45

(m, 4H), 3.21-3.12 (m, 1H), 2.99-2.89 (m, 8H), 2.86-2.70 (m,

3H), 2.19-2.07 (m, 3H), 1.72-1.57 (m, 2H).

D78
LCMS (ESI) m/z:

¹H NMR (400 MHz, DMSO-d₆) δ 11.09 (s, 1H), 8.66 (d, J = 7.6 Hz,

[M + H]⁺ = 881.50.
1H), 8.41 (s, 1H), 8.23 (s, 1H, FA), 7.95 (s, 1H), 7.68 (d, J = 8.5

Hz, 1H), 7.36 (d, J = 2.3 Hz, 1H), 7.27 (dd, J = 8.7, 2.3 Hz, 1H),

6.91 (s, 2H), 5.08 (dd, J = 12.9, 5.4 Hz, 1H), 4.35 (d, J = 13.1 Hz,

1H), 4.05-3.92 (m, 3H), 3.85 (s, 6H), 3.62 (s, 3H), 3.57 (s, 2H),

3.47-3.41 (m, 6H), 3.17-3.11 (m, 1H), 2.94-2.83 (m, 1H), 2.76-

2.66 (m, 1H), 2.61-2.55 (m, 7H), 2.22 (s, 6H), 2.06-1.98 (m,

1H), 1.92-1.79 (m, 2H), 1.55-1.32 (m, 2H).

D79
LCMS (ESI) m/z:

¹H NMR (300 MHz, Methanol-d4) δ 8.35 (d, J = 1.6 Hz, 1H), 7.88

[M + H]⁺ = 889.25
(s, 1H), 7.81 (d, J = 8.4 Hz, 1H), 7.51 (d, J = 2.2 Hz, 1H), 7.40 (d,

J = 8.4 Hz, 1H), 7.08 (s, 2H), 5.13 (dd, J = 12.1, 5.3 Hz, 1H), 4.58-

4.45 (m, 1H), 4.42 (s, 2H), 4.02 (s, 6H), 3.87-3.77 (m, 4H), 3.76

(s, 3H), 3.69-3.57 (m, 4H), 3.49-3.38 (m, 3H), 3.16 (d, J = 11.8

Hz, 1H), 3.03-2.97 (m, 2H), 2.92 (s, 6H), 2.90-2.70 (m, 3H),

2.70-2.41 (m, 2H), 2.19-1.94 (m, 3H).

D80
LCMS (ESI) m/z:

¹H NMR (400 MHz, Methanol-d4) δ 8.53 (s, 2H, FA), 8.32 (s, 1H),

[M + H]⁺ = 841.35
7.87 (s, 1H), 7.80 (dd, J = 8.6, 7.3 Hz, 1H), 7.47 (dd, J = 7.9, 4.8

Hz, 2H), 7.06 (s, 2H), 5.09 (dd, J = 12.7, 5.5 Hz, 1H), 4.41-4.31

(m, 4H), 4.23-4.17 (m, 1H), 4.02 (s, 6H), 3.99-3.82 (m, 1H),

3.75 (s, 3H), 3.49-3.38 (m, 2H), 2.89 (s, 6H), 2.88-2.82 (m,

4H), 2.78-2.65 (m, 2H), 2.23-2.12 (m, 3H), 2.06-1.93 (m, 8H),

1.86-1.75 (m, 2H).

D81
LCMS (ESI) m/z:

¹H NMR (300 MHz, DMSO-d6) δ 11.09 (s, 1H), 8.79 (s, 1H), 8.37

[M + H]⁺ = 828.30.
(s, 1H), 8.23 (s, 2H FA), 7.94 (s, 1H), 7.81 (dd, J = 8.5, 7.2 Hz,

1H), 7.53 (d, J = 8.5 Hz, 1H), 7.44 (d, J = 7.2 Hz, 1H), 6.91 (s, 2H),

5.08 (dd, J = 12.8, 5.4 Hz, 1H), 4.23 (t, J = 6.1 Hz, 2H), 3.84 (s,

6H), 3.62 (s, 3H), 3.53 (s, 2H), 3.17-3.11 (m, 3H), 2.93-2.88

(m, 2H), 2.64-2.59 (m, 2H), 2.42-2.31 (m, 3H), 2.19 (s, 6H),

2.07-1.98 (m, 1H), 1.95-1.84 (m, 2H), 1.81-1.74 (m, 2H), 1.71-

1.54 (m, 5H), 1.30-1.08 (m, 3H).

D82
LCMS (ESI) m/z:

¹H NMR (300 MHz, Methanol-d4) δ 8.56 (s, 2H, FA), 8.27 (s, 1H),

[M + H]+ = 838.98.
7.87-7.77 (m, 2H), 7.47 (dd, J = 7.7, 2.6 Hz, 2H), 7.03 (s, 2H),

5.21-5.10 (m, 1H), 4.30 (t, J = 5.7 Hz, 2H), 4.24 (s, 2H), 4.17-

4.08 (m, 1H), 4.00 (s, 7H), 3.75 (s, 3H), 3.29-3.14 (m, 2H), 3.10-

2.98 (m, 2H), 2.95-2.86 (m, 1H), 2.81-2.69 (m, 9H), 2.50 (dd,

J = 15.0, 8.0 Hz, 1H), 2.21-2.11 (m, 1H), 2.04-1.80 (m, 7H),

1.74-1.63 (m, 2H).

D83
LCMS (ESI) m/z:

¹H NMR (300 MHz, DMSO-d₆) δ 11.11 (s, 1H), 9.40 (s, 1H, TFA),

[M + H]⁺ = 863.
8.63 (s, 1H), 8.45 (s, 1H), 8.01 (s, 1H), 7.86 (d, J = 8.3 Hz, 1H),

7.45 (d, J = 2.3 Hz, 1H), 7.38 (dd, J = 8.3, 2.3 Hz, 1H), 7.02 (s,

2H), 5.13 (dd, J = 12.8, 5.4 Hz, 1H), 4.26 (dd, J = 9.4, 5.4 Hz, 4H),

3.93 (s, 7H), 3.63 (s, 4H), 3.40 (d, J = 1.3 Hz, 1H), 3.02-2.82 (m,

3H), 2.76 (d, J = 4.7 Hz, 7H), 2.66-2.53 (m, 2H), 2.53-2.52 (m,

1H), 2.26-1.90 (m, 7H), 1.77 (s, 1H), 1.60 (s, 1H).

D84
LCMS (ESI) m/z:

¹H NMR (400 MHz, DMSO-d6) δ 11.10 (s, 1H), 8.82 (t, J = 8.0 Hz,

[M + H]+ = 845.40.
1H), 8.48 (d, J = 67.2 Hz, 1H), 8.19 (s, 1H, FA), 7.96 (d, J = 2.3

Hz, 1H), 7.82 (t, J = 7.8 Hz, 1H), 7.53 (d, J = 8.5 Hz, 1H), 7.45 (d,

J = 7.2 Hz, 1H), 6.93 (s, 2H), 6.57 (s, 1H), 5.09 (dd, J = 12.9, 5.5

Hz, 1H), 4.81-4.45 (m, 1H), 4.23 (t, J = 6.4 Hz, 2H), 3.86 (s, 6H),

3.65-3.61 (m, 4H), 3.15-3.08 (m, 1H), 2.95-2.78 (m, 3H), 2.62-

2.57 (m, 2H), 2.34-2.25 (m, 7H), 2.20-1.92 (m, 4H), 1.90-

1.73 (m, 3H), 1.66-1.36 (m, 6H).

D85
LCMS (ESI) m/z:

¹H NMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 8.85 (d, J = 8.0 Hz,

[M + H]⁺ = 845.35
1H), 8.50 (d, J = 66.8 Hz, 1H), 8.14 (s, 1H, FA), 8.00 (d, J = 1.1

Hz, 1H), 7.85 (d, J = 8.2 Hz, 1H), 7.44 (d, J = 2.3 Hz, 1H), 7.36

(dd, J = 8.3, 2.2 Hz, 1H), 7.03 (s, 2H), 6.53 (s, 1H), 5.13 (dd, J =

13.0, 5.3 Hz, 1H), 4.27-4.17 (m, 4H), 3.93 (s, 7H), 3.64 (d, J =

1.5 Hz, 3H), 2.96-2.84 (m, 2H), 2.76 (s, 6H), 2.63-2.58 (m, 2H),

2.47-2.34 (m, 2H), 2.12-1.95 (m, 3H), 1.89-1.68 (m, 3H), 1.66-

1.40 (m, 5H), 1.30-1.21 (m, 1H).

D86
LCMS (ESI) m/z:

¹H NMR (300 MHz, Methanol-d4) δ 8.31 (br s, 1H, FA), 8.27 (s,

[M + H]+ = 881.45.
1H), 7.88 (s, 1H), 7.73 (d, J = 8.5 Hz, 1H), 7.42 (s, 1H), 7.30 (d,

J = 8.5 Hz, 1H), 7.06 (s, 2H), 5.10 (dd, J = 12.4, 5.3 Hz, 1H), 4.42 (s,

2H), 4.22-4.12 (m, 1H), 4.02 (s, 6H), 3.75 (s, 3H), 3.69-3.53

(m, 7H), 3.22-3.08 (m, 4H), 2.92 (s, 9H), 2.82-2.69 (m, 5H),

2.26 (d, J = 14.0 Hz, 2H), 2.16-1.95 (m, 3H), 1.93-1.81 (m, 2H),

1.81-1.67 (m, 2H).

D87
LCMS (ESI) m/z:

¹H NMR (300 MHz, DMSO-d6) δ 11.11 (s, 1H), 8.61 (d, J = 7.9 Hz,

[M + H]+ = 841.35.
1H), 8.39 (s, 1H), 8.18 (s, 1H, FA), 7.93 (s, 1H), 7.82 (d, J = 8.3

Hz, 1H), 7.42 (d, J = 2.2 Hz, 1H), 7.34 (d, J = 8.4 Hz, 1H), 6.91 (s,

2H), 5.11 (dd, J = 13.0, 5.3 Hz, 1H), 4.19 (t, J = 6.4 Hz, 2H), 4.03

(s, 1H), 3.85 (s, 6H), 3.64-3.56 (m, 5H), 2.75-2.72 (m, 1H),

2.69-2.60 (m, 3H), 2.60-2.53 (m, 4H), 2.30-2.22 (m, 7H), 1.97-

1.66 (m, 8H), 1.60-1.43 (m, 5H).

D88
LCMS (ESI) m/z:

¹H NMR (400 MHz, DMSO-d6) δ 11.10 (s, 1H), 8.60 (d, J = 7.9 Hz,

[M + H]⁺ = 841.35.
1H), 8.36 (d, J = 2.4 Hz, 1H), 8.19 (s, 2H, FA), 7.92 (s, 1H), 7.87-

7.74 (m, 1H), 7.51 (d, J = 9.0 Hz, 1H), 7.42 (dd, J = 7.2, 2.6 Hz,

1H), 6.90 (s, 2H), 5.11-5.04 (m, 1H), 4.23 (t, J = 6.3 Hz, 2H),

4.03 (s, 1H), 3.85 (s, 6H), 3.61 (s, 3H), 3.55 (s, 2H), 2.93-2.84

(m, 1H), 2.78-2.70 (m, 1H), 2.69-2.54 (m, 7H), 2.21 (s, 6H),

2.08-1.99 (m, 1H), 1.93-1.70 (m, 7H), 1.59-1.47 (m, 5H).

D89
LCMS (ESI) m/z:

¹H NMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 8.61 (d, J = 7.7 Hz,

[M + H]+ = 827.35.
1H), 8.42 (s, 1H), 8.21 (s, 1H, FA), 7.94 (s, 1H), 7.84 (d, J = 8.3

Hz, 1H), 7.44 (d, J = 2.3 Hz, 1H), 7.36 (dd, J = 8.3, 2.3 Hz, 1H),

6.91 (s, 2H), 5.12 (dd, J = 13.0, 5.4 Hz, 1H), 4.19 (t, J = 6.5 Hz,

2H), 3.84 (s, 6H), 3.74-3.67 (m, 1H), 3.62 (s, 3H), 3.55 (s, 2H),

2.95-2.84 (m, 3H), 2.64-2.54 (m, 2H), 2.35-2.29 (m, 2H), 2.21

(s, 6H), 2.07-1.94 (m, 3H), 1.83-1.73 (m, 4H), 1.62-1.42 (m,

6H).

D90
LCMS (ESI) m/z:

¹H NMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 8.88 (d, J = 8.8 Hz,

[M + H]+ = 863.35..
1H), 8.60 (s, 1H), 8.15 (s, 0.3H, FA), 7.98 (s, 1H), 7.84 (d, J = 8.3

Hz, 1H), 7.44 (d, J = 2.3 Hz, 1H), 7.36 (dd, J = 8.3, 2.3 Hz, 1H),

6.97 (s, 2H), 6.53 (s, 0.3H, FA salt), 5.13 (dd, J = 12.9, 5.3 Hz,

1H), 4.35 (s, 1H), 4.20 (t, J = 6.5 Hz, 2H), 3.88 (s, 6H), 3.82 (s,

2H), 3.63 (s, 3H), 3.15 (s, 1H), 2.95-2.84 (m, 2H), 2.65-2.55

(m, 3H), 2.44 (s, 8H), 2.18 (t, J = 11.3 Hz, 1H), 2.10-2.01 (m,

1H), 1.91-1.74 (m, 4H), 1.57-1.41 (m, 4H).

Example 30—Preparation of Compounds D91-D220, DD1, and DD2

In analogy to the procedures described in the examples above, compounds D91-D220, DD1, and DD2 were prepared using the appropriate starting materials

Compound

No.
LCMS

¹H NMR

D91
662.30

¹H NMR (400 MHz, Methanol-d4) δ 8.27-8.21 (m, 1H), 7.92 (s,

1H), 7.74-7.70 (m, 1H), 7.65-7.60 (m, 1H), 7.56-7.50 (m, 1H),

6.99 (s, 2H), 5.13 (dd, J = 13.3, 5.2 Hz, 1H), 4.53-4.38 (m, 2H),

4.18-4.10 (m, 2H), 4.10-4.01 (m, 2H), 3.97 (s, 6H), 3.83-3.74

(m, 2H), 3.71 (s, 3H), 3.67-3.57 (m, 1H), 2.99-2.85 (m, 1H),

2.84-2.75 (m, 1H), 2.56-2.40 (m, 1H), 2.26-2.15 (m, 1H).

D92
637.35

¹H NMR (300 MHz, Methanol-d4) δ 7.82-7.78 (m, 1H), 7.73 (s,

1H), 7.70-7.64 (m, 2H), 7.60-7.52 (m, 2H), 7.05 (s, 2H), 5.16

(dd, J = 13.3, 5.2 Hz, 1H), 4.58-4.42 (m, 2H), 4.27 (s, 2H), 4.24-

4.15 (m, 2H), 4.00 (s, 6H), 3.97-3.89 (m, 2H), 3.80-3.70 (m,

4H), 3.03-2.75 (m, 2H), 2.60-2.41 (m, 1H), 2.29-2.16 (m, 1H).

D93
674.35

¹H NMR (300 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.68 (d, J = 7.7 Hz,

1H), 8.40 (s, 1H), 7.97 (s, 1H), 7.46 (d, J = 8.4 Hz, 1H), 7.38-

7.22 (m, 3H), 7.19 (t, J = 1.9 Hz, 1H), 7.15-7.06 (m, 1H), 5.11

(dd, J = 13.2, 5.1 Hz, 1H), 4.41-4.18 (m, 2H), 4.06-3.92 (m,

3H), 3.80-3.76 (m, 3H), 3.60 (s, 3H), 3.02-2.83 (m, 3H), 2.60

(d, J = 16.5 Hz, 1H), 2.48-2.31 (m, 1H), 1.96 (dd, J = 27.3, 11.8

Hz, 3H), 1.70 (d, J = 12.2 Hz, 2H).

D94
808.55

¹H NMR (300 MHz, Methanol-d4) δ 7.53-7.48 (m, 2H), 7.40-

7.33 (m, 2H), 7.01-6.98 (m, 2H), 6.28 (s, 1H), 5.15 (dd, J = 13.3,

5.1 Hz, 1H), 4.51-4.42 (m, 2H), 4.41-4.35 (m, 2H), 4.06-3.96

(m, 10H), 3.71 (s, 3H), 3.49-3.40 (m, 4H), 3.25-3.07 (m, 6H),

3.02-2.85 (m, 3H), 2.84-2.76 (m, 1H), 2.60-2.51 (m, 1H), 2.50-

2.40 (m, 2H), 2.25-2.14 (m, 1H), 2.11-1.97 (m, 2H), 1.83-

1.48 (m, 5H).

D95
605.36

¹H NMR (400 MHz, DMSO-d6) δ 10.96 (s, 1H), 10.49 (s, 1H), 8.67

(s, 1H), 8.23-8.16 (m, 1H), 8.00 (s, 1H), 7.52 (t, J = 2.2 Hz, 1H),

7.38 (dd, J = 7.8, 1.9 Hz, 1H), 7.27 (t, J = 8.2 Hz, 1H), 6.94 (s, 2H),

6.80 (dd, J = 8.0, 2.5 Hz, 1H), 5.17 (dd, J = 10.7, 5.2 Hz, 1H), 3.86

(s, 6H), 3.64 (s, 3H), 3.63-3.55 (m, 2H), 2.81-2.70 (m, 1H), 2.68-

2.56 (m, 1H), 2.32-2.10 (m, 8H).

D96
631.35

¹H NMR (300 MHz, DMSO-d6) δ 10.96 (s, 1H), 8.72 (d, J = 7.7 Hz,

1H), 8.46 (s, 1H), 7.96 (s, 1H), 7.18 (t, J = 8.4 Hz, 1H), 6.86 (d, J =

2.2 Hz, 2H), 6.70-6.61 (m, 3H), 6.51 (d, J = 8.0 Hz, 1H), 5.26

(dd, J = 10.4, 5.2 Hz, 1H), 4.01 (s, 1H), 3.89 (s, 6H), 3.82 (d, J =

12.1 Hz, 2H), 3.67 (s, 3H), 2.88 (q, J = 10.2, 8.1 Hz, 2H), 2.81-

2.62 (m, 2H), 2.31-2.12 (m, 2H), 1.94 (d, J = 12.2 Hz, 2H), 1.81-

1.64 (m, 2H).

D97
711.20

¹H NMR (400 MHz, Methanol-d4) δ 8.54 (s, 1H, FA), 7.43 (s, 1H),

7.37 (d, J = 8.2 Hz, 1H), 7.01-6.91 (m, 3H), 6.85 (s, 1H), 6.77 (d,

J = 8.3 Hz, 1H), 6.29 (s, 1H), 5.16-5.08 (m, 1H), 4.44-4.30 (m,

2H), 4.18-3.99 (m, 2H), 3.98-3.92 (m, 6H), 3.75-3.67 (m, 7H),

3.62 (s, 1H), 3.08-2.95 (m, 2H), 2.88 (s, 3H), 2.82-2.70 (m,

1H), 2.50-2.39 (m, 1H), 2.22-2.11 (m, 1H), 2.05-2.00 (m, 4H).

D98
789.2

¹H NMR (400 MHz, DMSO-d6) δ 10.97 (s, 1H), 8.86 (s, 1H), 8.57

(s, 1H, FA), 7.92 (s, 1H), 7.43 (d, J = 8.3 Hz, 1H), 7.28 (d, J = 8.5

Hz, 1H), 7.23-7.05 (m, 2H), 6.79 (d, J = 2.3 Hz, 2H), 6.60-6.50

(m, 1H), 5.10 (dd, J = 13.2, 5.1 Hz, 1H), 4.42-4.12 (m, 3H), 3.90-

3.73 (m, 8H), 3.60 (s, 3H), 3.24-3.09 (m, 1H), 3.00-2.84 (m,

2H), 2.82-2.69 (m, 2H), 2.64-2.54 (m, 2H), 2.46-2.32 (m, 2H),

2.27-2.07 (m, 1H), 2.03-1.95 (m, 1H), 1.93-1.72 (m, 4H), 1.66-

1.49 (m, 2H).

D99
779.3

¹H NMR (300 MHz, DMSO-d6) δ 10.97 (s, 1H), 8.16 (s, FA, 1H),

7.88 (s, 1H), 7.70 (d, J = 5.4 Hz, 1H), 7.61 (d, J = 5.3 Hz, 1H), 7.38

(d, J = 8.2 Hz, 1H), 6.97 (s, 2H), 6.73-6.61 (m, 2H), 5.08 (dd, J =

13.3, 5.1 Hz, 1H), 4.36-4.14 (m, 2H), 3.88 (s, 6H), 3.81 (s, 2H),

3.62 (s, 3H), 3.58 (s, 4H), 3.13-3.02 (m, 2H), 2.99-2.78 (m,

2H), 2.64-2.55 (m, 1H), 2.47-2.38 (m, 2H), 2.38-2.20 (m, 4H),

2.19-2.06 (m, 2H), 2.03-1.92 (m, 1H), 1.87-1.53 (m, 7H), 1.32-

1.06 (m, 2H).

D100
809.55

¹H NMR (300 MHz, Methanol-d4) δ 7.66 (s, 1H), 7.41 (d, J = 8.2

Hz, 1H), 7.01 (s, 2H), 6.88 (s, 1H), 6.83-6.78 (m, 1H), 6.76 (s,

1H), 5.14 (dd, J = 13.2, 5.2 Hz, 1H), 4.56-4.30 (m, 4H), 4.04-

3.97 (m, 9H), 3.86-3.76 (m, 3H), 3.75-3.71 (m, 4H), 3.69-3.53

(m, 4H), 3.36-3.34 (m, 1H), 3.23-3.07 (m, 5H), 2.99-2.78 (m,

2H), 2.62-2.43 (m, 1H), 2.37-1.99 (m, 9H), 1.72-1.61 (m, 1H).

D101
804.55

¹H NMR (300 MHz, Methanol-d4) δ 8.61 (s, 1H) FA, 8.36 (s, 1H),

8.04 (s, 1H), 7.46 (d, J = 8.2 Hz, 1H), 7.10 (s, 2H), 6.94-6.88 (m,

1H), 6.87-6.81 (m, 1H), 5.20 (dd, J = 13.2, 5.1 Hz, 1H), 4.54-

4.39 (m, 4H), 4.08 (s, 6H), 3.81 (s, 3H), 3.76-3.72 (m, 4H), 3.65-

3.55 (m, 2H), 3.26-3.11 (m, 2H), 3.02-2.81 (m, 2H), 2.77-

2.40 (m, 7H), 2.31-2.18 (m, 1H), 2.16-2.07 (m, 2H), 2.00 (s,

5H), 1.72-1.49 (m, 2H).

D102
874.5

¹H NMR (400 MHz, DMSO-d6) δ 10.95 (s, 1H), 8.88 (d, J = 8.9 Hz,

1H), 8.59 (s, 1H), 8.17 (s, 2H, FA), 7.96 (s, 1H), 7.50 (d, J = 8.7 Hz,

1H), 7.05 (d, J = 7.9 Hz, 2H), 6.93 (s, 2H), 5.05 (dd, J = 13.3, 5.1

Hz, 1H), 4.38-4.16 (m, 3H), 3.85 (s, 8H), 3.63 (s, 5H), 3.19-3.14

(m, 2H), 2.95-2.76 (m, 5H), 2.64-2.58 (m, 1H), 2.41-2.36 (m,

2H), 2.26 (d, J = 4.0 Hz, 6H), 2.21-2.15 (m, 1H), 2.00-1.86 (m,

2H), 1.82-1.73 (m, 3H), 1.54-1.50 (m, 1H), 1.46-1.38 (m, 2H),

1.30-1.19 (m, 2H).

D103
650.2

¹H NMR (300 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.42 (d, 1H), 8.46

(s, 1H), 8.06-7.94 (m, 3H), 7.86-7.73 (m, 2H), 7.42 (d, 1H), 6.82-

6.72 (m, 2H), 5.11 (dd, 1H), 4.91-4.78 (m, 1H), 4.42-4.15 (m,

4H), 3.89-3.79 (m, 2H), 3.61 (s, 3H), 3.01-2.83 (m, 1H), 2.66-

2.53 (m, 1H), 2.45-2.28 (m, 1H), 2.05-1.95 (m, 1H).

D104
927.45

¹H NMR (400 MHz, DMSO-d6) δ 10.95 (s, 1H), 10.14-9.84 (m,

2H, TFA), 9.07 (s, 1H), 8.61 (s, 1H), 8.02 (s, 1H), 7.52 (d, J = 8.3

Hz, 1H), 7.03 (d, J = 2.9 Hz, 2H), 6.54-6.45 (m, 2H), 5.05 (dd,

J = 13.2, 5.2 z, 1H), 4.71-4.57 (m, 1H), 4.43-4.33 (m, 2H), 4.31-

4.16 (m, 4H), 4.08-4.00 (m, 2H), 3.93 (s, 6H), 3.78 (s, 2H), 3.69

(s, 3H), 3.51 (s, 5H), 3.45 (s, 2H), 3.39 (s, 2H), 3.19 (s, 2H), 3.01-

2.89 (m, 3H), 2.80 (s, 3H), 2.70-2.54 (m, 1H), 2.38-2.32 (m,

1H), 2.10 (s, 4H), 1.94 (d, J = 14.0 Hz, 3H).

D105
688.3

¹H NMR (300 MHz, DMSO-d6) δ 10.95 (s, 1H), 9.07 (s, 1H, TFA),

8.84-8.76 (m, 1H), 8.48 (s, 1H), 8.01 (s, 1H), 7.40-7.18 (m, 2H),

7.09-7.02 (m, 4H), 5.21-5.13 (m, 1H), 4.26 (d, J = 5.0 Hz, 2H),

4.15-4.01 (m, 2H), 3.93 (s, 6H), 3.64 (s, 3H), 3.30-3.14 (m, 2H),

2.78 (d, J = 4.5 Hz, 6H), 2.75-2.71 (m, 1H), 2.70-2.61 (m, 2H),

2.23-2.13 (m, 2H), 2.06-1.97 (m, 2H), 1.93-1.82 (m, 2H).

D106
696.3

¹H NMR (300 MHz, Methanol-d4) δ 7.78-7.67 (m, 2H), 7.60 (d,

J = 5.4 Hz, 1H), 7.44 (d, J = 8.3 Hz, 1H), 7.37-7.28 (m, 2H), 7.04

(s, 2H), 5.15 (dd, J = 13.3, 5.1 Hz, 1H), 4.49-4.32 (m, 2H), 4.20

(s, 2H), 3.98 (s, 6H), 3.76 (s, 3H), 3.60 (s, 1H), 3.47 (d, J = 10.6

Hz, 2H), 3.25-3.12 (m, 3H), 3.00-2.80 (m, 4H), 2.51 (dd, J =

12.9, 4.8 Hz, 1H), 2.24-2.03 (m, 5H), 1.84 (s, 2H).

D107
825.4

¹H NMR (400 MHz, DMSO-d6) δ 8.15 (s, 1H, FA), 7.86 (s, 1H),

7.68 (d, J = 5.4 Hz, 1H), 7.64 (d, J = 8.3 Hz, 1H), 7.60 (d, J = 5.4

Hz, 1H), 6.93 (s, 2H), 6.78 (s, 1H), 6.65 (dd, J = 8.4, 2.2 Hz, 1H),

5.78-5.68 (m, 1H), 5.63 (dd, J = 9.5, 4.0 Hz, 1H), 5.28-5.19 (m,

1H), 3.85 (s, 6H), 3.74 (s, 4H), 3.61 (s, 3H), 3.56 (s, 2H), 3.51 (s,

1H), 3.22-3.16 (m, 2H), 3.12-2.98 (m, 2H), 2.87-2.78 (m, 1H),

2.65-2.54 (m, 2H), 2.47-2.38 (m, 2H), 2.11-2.03 (m, 1H), 1.87-

1.80 (m, 1H), 1.76-1.72 (m, 4H), 0.89-0.83 (m, 3H), 0.83-

0.77 (m, 3H).

D108
688.35

¹H NMR (300 MHz, Methanol-d4) δ 8.56 (s, 1H, FA salt), 8.24 (s,

1H), 7.84 (s, 1H), 7.17 (t, J = 8.1 Hz, 1H), 7.05 (s, 2H), 6.74-6.64

(m, 2H), 6.57 (d, J = 8.2 Hz, 1H), 5.09 (dd, J = 9.6, 4.8 Hz, 1H),

4.34 (s, 2H), 4.01 (s, 7H), 3.74 (s, 5H), 2.94-2.88 (m, 1H), 2.85

(s, 6H), 2.81-2.71 (m, 3H), 2.43-2.13 (m, 2H), 2.10-2.00 (m,

2H), 1.85-1.75 (m, 2H).

D109
670.2

¹H NMR (300 MHz, DMSO-d6) δ 8.36 (s, 1H), 7.86 (s, 1H), 7.48

(d, J = 8.4 Hz, 1H), 7.38-7.32 (m, 1H), 7.29 (s, 1H), 6.77 (d, J =

2.2 Hz, 2H), 6.56 (t, J = 2.2 Hz, 1H), 5.07 (dd, J = 13.2, 5.1 Hz,

1H), 4.42-4.17 (m, 2H), 3.98 (s, 1H), 3.82-3.71 (m, 8H), 3.58 (s,

3H), 3.04-2.80 (m, 3H), 2.61 (d, J = 17.7 Hz, 1H), 2.41-2.30 (m,

1H), 2.05-1.87 (m, 3H), 1.71 (dd, J = 22.0, 10.0 Hz, 2H).

D110
724.3

¹H NMR (300 MHz, Methanol-d4) δ 8.57 (s, 1H, FA salt), 7.68 (s,

1H), 7.44-7.35 (m, 2H), 7.00 (s, 2H), 6.87 (d, J = 2.1 Hz, 1H),

6.79 (dd, J = 8.2, 2.3 Hz, 1H), 5.14 (dd, J = 13.1, 5.1 Hz, 1H), 4.48-

4.30 (m, 2H), 4.14 (s, 2H), 3.98 (s, 6H), 3.73 (s, 7H), 3.33-3.20

(m, 1H), 3.08 (s, 4H), 2.99-2.73 (m, 2H), 2.58-2.41 (m, 1H),

2.17 (d, J = 12.6 Hz, 1H), 2.07 (s, 4H), 1.41 (d, J = 6.8 Hz, 6H).

D111
718.35

¹H NMR (400 MHz, DMSO-d6) δ 10.97 (s, 1H), 7.89 (s, 1H), 7.74-

7.55 (m, 2H), 7.39 (d, J = 8.8 Hz, 1H), 6.95 (s, 2H), 6.77-6.67 (m,

2H), 5.17-5.00 (m, 1H), 4.42-4.11 (m, 2H), 3.87 (s, 8H), 3.72-

3.55 (m, 7H), 3.02-2.81 (m, 1H), 2.75-2.54 (m, 4H), 2.46-2.28

(m, 2H), 2.00 (s, 3H).

D112
875.45

¹H NMR (300 MHz, Methanol-d4) δ 8.42 (d, J = 15.6 Hz, 1H), 7.87

(s, 1H), 7.55 (d, J = 8.5 Hz, 1H), 7.41 (d, J = 7.6 Hz, 2H), 7.08 (s,

2H), 5.27-5.10 (m, 1H), 4.44 (d, J = 14.5 Hz, 5H), 4.02 (s, 6H),

3.75 (s, 3H), 3.63 (s, 8H), 3.46 (s, 3H), 3.14 (d, J = 12.3 Hz, 1H),

3.01-2.89 (m, 9H), 2.90-2.77 (m, 1H), 2.78-2.38 (m, 3H), 2.26-

2.16 (m, 1H), 2.10-1.91 (m, 2H).

D113
696.3

¹H NMR (300 MHz, DMSO-d6) δ 8.32 (s, 1H, FA), 7.80 (s, 1H),

7.69-7.56 (m, 2H), 7.43 (d, J = 8.2 Hz, 1H), 7.28-7.13 (m, 2H),

7.01-6.88 (m, 2H), 5.04-4.89 (m, 1H), 4.31 (d, 2H), 4.19 (d, J =

16.2 Hz, 2H), 3.91-3.72 (m, 6H), 3.60 (s, 5H), 3.48 (s, 1H), 3.33-

2.91 (m, 4H), 2.90-2.71 (m, 2H), 2.71-2.56 (m, 1H), 2.43-

2.31 (m, 1H), 2.29-2.12 (m, 1H), 2.11-1.80 (m, 4H), 1.70 (d, J =

13.8 Hz, 1H), 1.48 (s, 1H).

D114
713.35

¹H NMR (300 MHz, DMSO-d6 + a drop of D2O) δ 8.45 (s, 1H), 7.97

(s, 1H), 7.41 (d, J = 8.3 Hz, 1H), 7.01 (s, 2H), 6.91-6.83 (m, 1H),

6.80 (d, J = 2.3 Hz, 1H), 5.06 (dd, J = 13.3, 5.1 Hz, 1H), 4.58 (t,

J = 5.7 Hz, 1H), 4.38-4.10 (m, 4H), 3.92 (s, 6H), 3.53-3.45 (m,

1H), 3.42-3.36 (m, 3H), 3.39-3.25 (m, 3H), 2.92-2.75 (m, 1H),

2.77 (s, 6H), 2.66-2.54 (m, 1H), 2.43-2.24 (m, 2H), 2.17-2.07

(m, 1H), 2.05-1.95 (m, 1H).

D115
642.3

¹H NMR (300 MHz, DMSO-d6) δ 10.99 (s, 1H), 9.39 (d, J = 7.1 Hz,

1H), 8.43 (s, 1H), 7.92 (s, 1H), 7.42 (d, J = 8.8 Hz, 1H), 6.78 (dd,

J = 6.6, 2.3 Hz, 4H), 6.58 (t, J = 2.2 Hz, 1H), 5.11 (dd, J = 13.2, 5.0

Hz, 1H), 4.84 (d, J = 6.8 Hz, 1H), 4.40-4.15 (m, 4H), 3.82 (s, 8H),

3.60 (s, 3H), 2.95-2.83 (m, 1H), 2.62 (s, 1H), 2.45-2.34 (m, 1H),

2.05-1.94 (m, 1H).

D116
669.35

¹H NMR (300 MHz, DMSO-d6) δ 10.99 (s, 1H), 9.41 (d, J = 7.1 Hz,

1H), 8.45 (s, 1H), 8.16 (s, 0.2H, FA), 7.98 (s, 1H), 7.43 (d, J = 8.8

Hz, 1H), 6.95 (s, 2H), 6.82-6.72 (m, 2H), 5.11 (dd, J = 13.2, 5.0

Hz, 1H), 4.89-4.79 (m, 1H), 4.40-4.15 (m, 4H), 3.87 (s, 6H), 3.84-

3.74 (m, 4H), 3.62 (s, 3H), 2.95-2.83 (m, 1H), 2.63 (s, 1H), 2.39

(s, 7H), 2.05-1.95 (m, 1H).

D117
718.3

¹H NMR (300 MHz, DMSO) δ 10.98 (s, 1H), 8.99 (d, J = 7.6 Hz,

1H), 8.43 (s, 1H), 8.05-7.94 (m, 3H), 7.86-7.73 (m, 2H), 7.44

(d, J = 8.3 Hz, 1H), 7.38-7.17 (m, 2H), 5.10 (dd, J = 13.3, 5.1 Hz,

1H), 4.48-4.40 (m, 1H), 4.39-4.18 (m, 2H), 3.62 (s, 3H), 3.31-

3.12 (m, 5H), 3.01-2.83 (m, 1H), 2.65-2.55 (m, 2H), 2.45-2.34

(m, 1H), 2.33-2.19 (m, 2H), 2.05-1.85 (m, 3H), 1.76-1.70 (m,

4H).

D118
751.35

¹H NMR (300 MHz, DMSO-d6) δ 10.99 (s, 1H), 8.98-8.89 (m,

1H), 8.59 (d, J = 2.3 Hz, 1H), 8.15 (s, 0.2H, FA salt), 8.02 (s, 1H),

7.46 (d, J = 8.5 Hz, 1H), 7.36 (dt, J = 8.6, 2.2 Hz, 1H), 7.27 (d, J =

2.3 Hz, 1H), 7.20-7.10 (m, 2H), 5.12 (dd, J = 13.2, 5.2, 1H), 4.70-

4.49 (m, 1H), 4.36 (d, J = 16.9 Hz, 1H), 4.29-4.08 (m, 2H), 3.91

(s, 3H), 3.87 (s, 1H), 3.62 (s, 5H), 3.51-3.42 (m, 1H), 3.21-3.08

(m, 1H), 3.01-2.83, (m, 1H), 2.66-2.55 (m, 1H), 2.45-2.37 (m,

1H), 2.27 (s, 6H), 2.08-1.92 (m, 3H).

D119
726.35

¹H NMR (300 MHz, Methanol-d4) δ 8.56 (s, 1H, FA salt), 7.75 (s,

1H), 7.59 (s, 1H), 7.41 (d, J = 8.2 Hz, 1H), 7.04 (s, 2H), 6.88 (s,

1H), 6.80 (d, J = 8.3 Hz, 1H), 5.19-5.08 (m, 1H), 4.72 (s, 2H),

4.40 (d, J = 5.2 Hz, 2H), 4.27 (s, 2H), 4.00 (s, 6H), 3.82-3.62 (m,

8H), 3.42 (s, 3H), 3.24-3.18 (m, 3H), 3.00-2.71 (m, 2H), 2.56-

2.45 (m, 1H), 2.15-2.09 (m, 5H).

D120
712.3

¹H NMR (300 MHz, Methanol-d4) δ 8.57 (s, 1H), 7.63 (s, 1H), 7.39

(d, J = 8.2 Hz, 1H), 6.93 (s, 2H), 6.85 (d, J = 2.2 Hz, 1H), 6.80-

6.73 (m, 2H), 5.13 (dd, J = 13.3, 5.1 Hz, 1H), 4.47-4.27 (m, 2H),

4.02 (s, 5H), 3.95 (s, 6H), 3.72 (s, 3H), 3.70 (s, 4H), 3.06-2.72 (m,

6H), 2.60-2.37 (m, 1H), 2.23-2.11 (m, 1H), 2.01 (t, J = 5.6 Hz,

4H).

D121
696.35

¹H NMR (300 MHz, DMSO-d6) δ 8.26 (s, 1H, FA), 7.83 (s, 1H),

7.67 (d, J = 5.4 Hz, 1H), 7.60 (d, J = 5.4 Hz, 1H), 7.50 (d, J = 8.4

Hz, 1H), 7.00 (d, J = 9.3 Hz, 2H), 6.91 (d, J = 1.9 Hz, 2H), 5.01

(dd, J = 13.3, 5.1 Hz, 1H), 4.31 (d, J = 16.9 Hz, 1H), 4.21 (d, 1H),

3.80 (d, J = 1.6 Hz, 7H), 3.71-3.60 (m, 6H), 3.50 (s, 1H), 3.28-

3.01 (m, 2H), 2.98-2.77 (m, 1H), 2.59 (d, J = 17.1 Hz, 4H), 2.39-

2.24 (m, 1H), 2.05-1.90 (m, 2H), 1.89-1.78 (m, 1H), 1.77-1.58

(m, 3H), 1.57-1.42 (m, 1H).

D122
613.25

¹H NMR (400 MHz, DMSO-d6) δ 10.97 (s, 1H), 7.86 (s, 1H), 7.69

(d, J = 5.4 Hz, 1H), 7.60 (d, J = 5.3 Hz, 1H), 7.39 (d, J = 8.3 Hz,

1H), 6.96 (s, 2H), 6.86 (dd, J = 8.4, 2.4 Hz, 1H), 6.79 (d, J = 2.3

Hz, 1H), 5.10 (dd, J = 13.3, 5.1 Hz, 1H), 4.33 (d, J = 16.4 Hz, 1H),

4.25-4.07 (m, 2H), 3.85 (s, 6H), 3.62 (s, 4H), 3.55-3.40 (m,

3H), 2.99-2.83 (m, 1H), 2.71-2.55 (m, 2H), 2.46-2.34 (m, 1H),

2.18-2.08 (m, 1H), 2.08-1.97 (m, 1H).

D123
627.2

¹H NMR (400 MHz, DMSO-d6 with a drop of D2O) δ 8.30 (s, 0.1H,

FA), 7.82 (s, 1H), 7.67 (d, J = 5.4 Hz, 1H), 7.60 (d, J = 5.4 Hz, 1H),

7.45 (d, J = 8.4 Hz, 1H), 7.35-7.28 (m, 1H), 7.23-7.18 (m, 1H),

6.90 (s, 2H), 5.08 (dd, J = 13.3, 5.1 Hz, 1H), 4.40-4.18 (m, 2H),

3.88-3.78 (m, 8H), 3.60 (s, 3H), 3.42-3.32 (m, 1H), 2.97-2.83

(m, 1H), 2.82-2.70 (m, 2H), 2.68-2.55 (m, 1H), 2.49-2.31 (m,

3H), 2.05-1.97 (m, 1H), 1.55 (d, J = 12.4 Hz, 2H).

D124
687.35

¹H NMR (400 MHz, DMSO-d6) δ 10.18 (s, 1H), 8.67 (d, J = 7.7 Hz,

1H), 8.41 (s, 1H), 8.22 (s, 1H, FA), 7.94 (s, 1H), 7.23-7.14 (m,

1H), 6.89 (d, J = 14.8 Hz, 4H), 6.67 (d, J = 7.4 Hz, 1H), 4.46 (s,

2H), 4.00-3.87 (m, 1H), 3.84 (s, 6H), 3.75 (s, 1H), 3.72 (s, 1H),

3.61 (s, 3H), 3.56 (s, 2H), 3.28 (t, J = 6.8 Hz, 2H), 2.81 (t, J = 11.9

Hz, 2H), 2.54 (d, J = 6.8 Hz, 2H), 2.22 (s, 6H), 1.93-1.85 (m, 2H),

1.72-1.59 (m, 2H).

D125
552.1

¹H NMR (400 MHz, DMSO-d6) δ 11.02 (s, 1H), 8.14-7.99 (m,

3H), 7.76-7.70 (m, 2H), 7.70-7.59 (m, 4H), 5.16 (dd, J = 13.3,

5.1 Hz, 1H), 4.57 (d, J = 17.5 Hz, 1H), 4.43 (d, J = 17.5 Hz, 1H),

3.63 (s, 3H), 3.00-2.87 (m, 1H), 2.70-2.58 (m, 1H), 2.45-2.32

(m, 1H), 2.11-2.01 (m, 1H).

D126
846.4

¹H NMR (400 MHz, DMSO-d6) δ 8.62 (d, J = 7.7 Hz, 1H), 8.41 (s,

1H), 8.26 (s, 1H, FA), 7.94 (s, 1H), 7.44 (d, J = 8.4 Hz, 1H), 7.32

(d, J = 8.4 Hz, 1H), 7.21 (s, 1H), 6.90 (s, 2H), 5.10 (dd, J = 13.3,

5.1 Hz, 1H), 4.39-4.16 (m, 2H), 4.01-3.97 (m, 1H), 3.90 (d, J =

12.3 Hz, 1H), 3.84 (s, 6H), 3.72 (d, J = 10.4 Hz, 1H), 3.78-3.56

(m, 3H), 3.52 (s, 2H), 3.07-3.02 (m, 1H), 2.98-2.82 (m, 5H),

2.64-2.55 (m, 3H), 2.52-2.28 (m, 1H), 2.19 (s, 6H), 2.03-1.87

(m, 3H), 1.82-1.77 (m, 2H), 1.58-1.51 (m, 2H).

D127
846.35

¹H NMR (400 MHz, DMSO-d6) δ 8.61 (d, J = 7.6 Hz, 1H), 8.41 (s,

1H), 8.20 (s, 1H, FA), 7.94 (s, 1H), 7.52 (d, J = 8.5 Hz, 1H), 7.17-

7.08 (m, 2H), 6.93 (s, 2H), 5.05 (dd, J = 13.2, 5.1 Hz, 1H), 4.45-

4.16 (m, 2H), 4.12 (s, 1H), 4.01 (d, J = 12.4 Hz, 1H), 3.86 (s, 6H),

3.78-3.65 (m, 3H), 3.62 (s, 3H), 3.15 (d, J = 12.9 Hz, 1H), 3.05-

2.82 (m, 5H), 2.64-2.55 (m, 3H), 2.42-2.34 (m, 1H), 2.32 (s,

6H), 2.01-1.87 (m, 3H), 1.81-1.77 (m, 2H), 1.58-1.50 (m, 2H).

D128
678.1

¹H NMR (300 MHz, DMSO-d6) δ 10.99 (s, 1H), 8.70 (d, J = 7.7 Hz,

1H), 8.43 (s, 1H), 8.08-7.91 (m, 3H), 7.80 (d, J = 6.7 Hz, 2H),

7.47 (d, J = 8.5 Hz, 1H), 7.31 (d, J = 25.7 Hz, 2H), 5.12 (dd, J =

13.2, 5.1 Hz, 1H), 4.44-4.15 (m, 2H), 4.11-3.90 (m, 1H), 3.82

(d, J = 12.5 Hz, 2H), 3.61 (s, 3H), 3.01-2.91 (m, 2H), 2.90 (s,

1H), 2.74 (s, 1H), 2.73-2.70 (m, 1H), 2.68-2.55 (m, 1H), 2.43-

231 (m, 1H), 2.07-1.85 (m, 3H), 1.80-1.61 (m, 2H).

D129
704.3

¹H NMR (400 MHz, DMSO-d6) δ 10.99 (s, 1H), 8.92 (dd, J = 9.0,

2.9 Hz, 1H), 8.57 (d, J = 2.9 Hz, 1H), 7.89 (s, 1H), 7.46 (d, J = 8.4

Hz, 1H), 7.36 (d, J = 8.5, 2.6 Hz, 1H), 7.26 (s, 1H), 7.09 (s, 1H),

7.01 (s, 1H), 6.87 (s, 1H), 5.17-5.07 (m, 1H), 4.62-4.57 (m,

1H), 4.36 (d, J = 16.8 Hz, 1H), 4.28-4.10 (m, 2H), 3.92-3.80 (m,

4H), 3.61 (s, 3H), 3.48-3.40 (m, 1H), 3.20-3.09 (m, 1H), 2.99-

2.86 (m, 1H), 2.72-2.56 (m, 3H), 2.44-2.36 (m, 1H), 2.13-1.92

(m, 3H), 1.24 (t, J = 7.6 Hz, 3H).

D130
839.4

¹H NMR (300 MHz, DMSO-d6) δ 11.00 (s, 1H), 9.26-8.96 TFA

(m, 1H), 8.83 (s, 1H), 8.47 (s, 1H), 8.02 (s, 1H), 7.49 (d, J = 8.3

Hz, 1H), 7.38-7.22 (m, 2H), 7.02 (s, 2H), 5.11 (dd, J = 13.3, 5.1

Hz, 1H), 4.45-4.11 (m, 5H), 4.09-3.76 (m, 9H), 3.67-3.61 (m,

5H), 3.28-3.06 (m, 6H), 3.04-2.85 (m, 3H), 2.84-2.71 (m, 6H),

2.69-2.56 (m, 2H), 2.46-2.26 (m, 2H), 2.23-1.92 (m, 4H), 1.92-

1.72 (m, 2H).

D131
706.25

¹H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.94-8.87 (m,

1H), 8.56 (d, J = 2.8 Hz, 1H), 7.92 (s, 1H), 7.46 (d, J = 8.4 Hz, 1H),

7.40-7.32 (m, 1H), 7.26 (d, J = 1.5 Hz, 1H), 6.79 (d, J = 2.2 Hz,

2H), 6.57 (t, J = 2.2 Hz, 1H), 5.15-5.07 (m, 1H), 4.70-4.51 (m,

1H), 4.35 (d, J = 16.8 Hz, 1H), 4.27-4.18 (m, 1H), 4.14 (s, 1H),

3.88 (d, J = 13.0 Hz, 1H), 3.82 (s, 6H), 3.60 (s, 3H), 3.47-3.40 (m,

1H), 3.15-3.11 (m, 1H), 2.94-2.86 (m, 1H), 2.64-2.56 m, 1H),

2.42-2.38 (m, 1H), 2.04-1.89 (m, 3H).

D132
696.35

¹H NMR (400 MHz, DMSO-d6) δ 7.76 (s, 1H), 7.38 (d, J = 8.1 Hz,

1H), 7.29 (d, J = 1.3 Hz, 1H), 6.89 (s, 2H), 6.73-6.66 (m, 2H),

5.02 (dd, J = 13.3, 5.1 Hz, 1H), 4.36-4.12 (m, 2H), 3.84 (s, 6H),

3.72-3.62 (m, 4H), 3.60-3.55 (m, 7H), 2.93-2.80 (m, 1H), 2.70-

2.52 (m, 6H), 2.52-2.25 (m, 1H), 2.43-2.27 (m, 1H), 2.03-

1.95 (m, 1H), 1.80-1.76 (m, 4H).

D133
753.3

¹H NMR (300 MHz, DMSO-d6) δ 10.95 (s, 1H), 8.20 (s, 1H, FA),

7.96 (s, 1H), 7.87 (s, 1H), 7.47 (d, J = 8.2 Hz, 1H), 6.91 (s, 2H),

6.54-6.43 (m, 2H), 5.04 (dd, J = 13.3, 5.1 Hz, 1H), 4.36-4.11

(m, 2H), 3.85 (s, 6H), 3.62 (s, 8H), 3.57-3.51 (m, 3H), 3.30-

3.25 (m, 2H), 3.08-3.02 (m, 2H), 2.99-2.78 (m, 1H), 2.64-2.58

(m, 2H), 2.45-2.24 (m, 4H), 2.00-1.92 (m, 1H), 1.76-1.70 (m,

4H).

D134
846.35

¹H NMR (400 MHz, Methanol-d4) δ 8.38 (s, 1H), 7.89 (s, 1H), 7.49

(d, J = 8.4 Hz, 1H), 7.41-7.34 (m, 2H), 7.07 (s, 2H), 5.16 (dd, J =

13.3, 5.1 Hz, 1H), 4.43 (d, J = 10.6 Hz, 4H), 4.02 (s, 6H), 4.00-

3.93 (m, 3H), 3.76 (s, 3H), 3.69-3.57 (m, 2H), 3.39 (s, 2H), 3.24

(d, J = 13.0 Hz, 1H), 2.92 (s, 9H), 2.85-2.81 (m, 1H), 2.55-2.46

(m, 1H), 2.30-2.15 (m, 5H), 1.99-1.87 (m, 2H).

D135
846.5

¹H NMR (400 MHz, Methanol-d4) δ 8.55 (s, 1H, FA), 8.37 (s, 1H),

7.86 (s, 1H), 7.47 (d, J = 8.3 Hz, 1H), 7.40-7.33 (m, 2H), 7.05 (s,

2H), 5.15 (d, J = 8.1 Hz, 1H), 4.49-4.41 (m, 3H), 4.35 (d, J = 13.8

Hz, 2H), 4.01 (s, 6H), 3.86 (d, J = 12.1 Hz, 2H), 3.76 (s, 3H), 3.37

(s, 1H), 3.16-3.10 (m, 1H), 2.98-2.91 (m, 1H), 2.89-2.79 (m,

9H), 2.71-2.61 (m, 2H), 2.57-2.49 (m, 2H), 2.25-2.17 (m, 1H),

2.07-1.97 (m, 4H), 1.81-1.68 (m, 2H).

D136
670.3

¹H NMR (400 MHz, DMSO-d6) δ 10.97 (s, 1H), 8.68 (s, TFA, 1H),

7.93 (s, 1H), 7.72 (d, J = 5.3 Hz, 1H), 7.66-7.57 (m, 2H), 7.20-

7.12 (m, 2H), 7.08 (s, 2H), 5.07 (dd, J = 13.2, 5.1 Hz, 1H), 4.50 (d,

J = 13.9, 4.3 Hz, 1H), 4.43-4.19 (m, 3H), 3.97 (s, 6H), 3.95-

3.74 (m, 4H), 3.73-3.62 (m, 4H), 3.61-3.53 (m, 1H), 2.99-2.85

(m, 1H), 2.64-2.57 (m, 1H), 2.45-2.36 (m, 1H), 2.04-1.93 (m,

1H), 1.50 (dd, J = 6.5, 3.3 Hz, 6H).

D137
858.6

¹H NMR (300 MHz, Methanol-d4) δ 8.36 (s, 1H), 7.90 (d, J = 1.6

Hz, 1H), 7.57 (d, J = 9.0 Hz, 1H), 7.08 (d, J = 1.3 Hz, 2H), 6.79 (d,

J = 7.2 Hz, 2H), 5.11 (dd, J = 13.4, 5.1 Hz, 1H), 4.45-4.31 (m,

6H), 4.12-3.90 (m, 9H), 3.88-3.81 (m, 1H), 3.79-3.66 (m, 4H),

3.62-3.54 (m, 2H), 3.03-2.85 (m, 8H), 2.84-2.74 (m, 1H), 2.55-

2.10 (m, 9H).

D138
739.45

¹H NMR (300 MHz, DMSO-d6) δ 10.99 (s, 1H), 8.97 (s, 1H, TFA),

8.81 (d, J = 4.9 Hz, 1H), 8.29 (s, 1H), 8.00 (s, 1H), 7.41 (d, J = 8.8

Hz, 1H), 7.04 (s, 2H), 6.71 (dd, J = 5.7, 2.4 Hz, 2H), 5.08 (dd, J =

13.3, 5.1 Hz, 1H), 4.39-4.13 (m, 4H), 3.95 (s, 6H), 3.76 (s, 2H),

3.64 (s, 4H), 3.38 (d, J = 12.2 Hz, 2H), 3.19-3.09 (m, 2H), 2.94-

2.82 (m, 1H), 2.79 (d, J = 4.2 Hz, 3H), 2.60 (d, J = 16.2 Hz, 1H),

2.43-2.25 (m, 1H), 2.14 (d, J = 13.7 Hz, 2H), 2.06-1.96 (m, 3H).

D139
707.35

¹H NMR (400 MHz, DMSO-d6) δ 10.97 (s, 1H), 8.49 (d, J = 1.6 Hz,

1H), 8.14 (d, J = 1.8 Hz, 2H), 7.39 (d, J = 8.0 Hz, 1H), 6.97 (s, 2H),

6.70 (d, J = 7.5 Hz, 2H), 5.08 (dd, J = 13.2, 5.1 Hz, 1H), 4.36-4.15

(m, 2H), 3.90 (s, 8H), 3.63 (d, J = 4.3 Hz, 8H), 2.96-2.85 (m, 2H),

2.77 (s, 2H), 2.63-2.60 (m, 1H), 2.38 (dd, J = 13.2, 4.6 Hz, 1H),

2.06-1.95 (m, 2H), 1.87 (s, 3H)

D140
846.25

¹H NMR (300 MHz, Acetonitrile-d3) δ 8.29 (s, 1H), 7.74 (s, 1H),

7.46 (d, J = 8.3 Hz, 1H), 7.39-7.27 (m, 2H), 6.98 (s, 2H), 5.07

(dd, J = 13.3, 5.2 Hz, 1H), 4.87-4.65 (m, 1H), 4.42-4.23 (m,

3H), 3.94 (s, 7H), 3.83 (s, 1H), 3.64 (d, J = 20.7 Hz, 4H), 3.55-

3.37 (m, 2H), 3.30-2.99 (m, 3H), 2.81-2.80 (m, 6H), 2.52-2.30

(m, 3H), 2.27-2.10 (m, 4H), 1.87 (s, 2H), 1.73 (s, 1H), 1.32 (d,

J = 21.7 Hz, 2H).

D141
846.2

¹H NMR (300 MHz, Acetonitrile-d3) δ 8.29 (s, 1H), 7.75 (s, 1H),

7.48 (d, J = 8.2 Hz, 1H), 7.41-7.30 (m, 2H), 6.97 (s, 2H), 5.07

(dd, J = 13.3, 5.1 Hz, 1H), 4.79 (d, J = 22.1 Hz, 1H), 4.32 (q, J =

10.1, 9.0 Hz, 3H), 3.93 (s, 7H), 3.67 (s, 4H), 3.62-3.48 (m, 3H),

2.95-2.90 (m, 4H), 2.80 (s, 6H), 2.43 (dd, J = 13.1, 5.0 Hz, 1H),

2.22 (q, J = 22.0, 16.5 Hz, 5H), 1.94-1.82 (m, 2H), 1.74 (s, 1H),

1.31 (d, J = 21.7 Hz, 2H).

D142
860.5

¹H NMR (400 MHz, DMSO-d6) δ 8.58 (d, J = 1.2 Hz, 1H), 8.28 (s,

1H, FA), 7.95 (s, 1H), 7.43 (d, J = 8.4 Hz, 1H), 7.28 (dd, J = 8.5,

2.4 Hz, 1H), 7.18 (d, J = 2.3 Hz, 1H), 6.92 (s, 2H), 5.16 (dd, J =

13.4, 5.1 Hz, 1H), 4.42-4.13 (m, 3H), 3.84 (s, 6H), 3.80 (s, 2H),

3.62 (s, 3H), 3.57 (s, 2H), 3.17 (s, 1H), 3.00 (s, 3H), 2.96 (dd, J =

12.8, 5.2 Hz, 2H), 2.80-2.69 (m, 3H), 2.62-2.58 (m, 2H), 2.43-

2.35 (m, 2H), 2.22 (s, 6H), 2.04-1.97 (m, 1H), 1.84-1.80 (m,

4H), 1.71-1.46 (m, 2H).

D143
777.55

¹H NMR (400 MHz, DMSO-d6 with a drop of D2O) δ 8.5-8.50 (m,

1H), 8.28 (s, 1H), 7.95 (s, 1H), 7.47 (d, J = 8.4 Hz, 1H), 7.38-7.31

(m, 1H), 7.26 (s, 1H), 6.97 (s, 2H), 5.12 (dd, J = 13.3, 4.9 Hz, 1H),

4.58-4.53 (m, 1H), 4.36 (d, J = 16.9 Hz, 1H), 4.22 (dd, J = 17.0,

3.6 Hz, 1H), 4.13 (s, 1H), 4.04 (s, 2H), 3.88 (s, 7H), 3.61 (s, 3H),

3.41-3.28 (m, 1H), 3.11 (t, J = 12.3 Hz, 1H), 2.99 (s, 3H), 2.97-

2.88 (m, 1H), 2.81-2.72 (m, 1H), 2.59 (s, 6H), 2.42-2.34 (m, 1H),

2.06-2.01 (m, 2H), 1.96-1.91 (m, 1H).

D144
670.2

¹H NMR (400 MHz, Methanol-d4) δ 7.77 (s, 1H), 7.71 (d, J = 5.4

Hz, 1H), 7.62 (d, J = 5.4 Hz, 1H), 7.53 (d, J = 8.2 Hz, 1H), 7.38 (d,

J = 10.2 Hz, 2H), 7.09 (s, 2H), 5.24-5.12 (m, 1H), 4.69-4.53 (m,

2H), 4.52-4.37 (m, 3H), 4.04 (s, 6H), 3.88 (s, 2H), 3.76 (s, 3H),

3.74-3.51 (m, 3H), 3.00-2.86 (m, 1H), 2.85-2.76 (m, 1H), 2.61-

2.42 (m, 1H), 2.25-2.15 (m, 1H), 1.62 (d, J = 6.4 Hz, 6H).

D145
858.6

¹H NMR (300 MHz, Methanol-d4) δ 8.57 (br s, 2H, FA), 8.36 (d,

J = 1.5 Hz, 1H), 7.88 (s, 1H), 7.35 (d, J = 8.4 Hz, 1H), 7.01 (d, J =

11.0 Hz, 4H), 5.15 (dd, J = 13.7, 6.0 Hz, 1H), 4.67-4.62 (m, 3H),

4.45-4.36 (m, 3H), 4.18 (s, 2H), 3.99 (s, 6H), 3.91-3.81 (m,

2H), 3.76 (s, 3H), 3.67 (s, 1H), 3.53 (s, 2H), 2.94-2.81 (m, 2H),

2.72 (s, 6H), 2.58-2.13 (m, 9H).

D146
727.3

¹H NMR (300 MHz, DMSO-d6) δ 10.99 (s, 1H), 8.68 (m, J = 7.7

Hz, 1H), 8.41 (s, 1H), 8.18 (d, J = 2.2 Hz, 1H), 7.44 (m, J = 8.5 Hz,

1H), 7.21 (m, J = 2.3 Hz, 1H), 6.95 (s, 2H), 5.11 (dd, J = 13.3, 5.1

Hz, 1H), 4.45-4.26 (m, 1H), 4.25-4.12 (m, 1H), 4.05-3.95 (m,

2H), 3.92 (s, 6H), 3.83-3.81 (m, 1H), 3.80-3.76 (m, 2H), 3.62

(s, 3H), 3.00-2.81 (m, 3H), 2.55 (s, 2H), 2.39 (s, 6H), 2.06-1.96

(m, 1H), 1.95-1.81 (m, 2H), 1.78-1.55 (m, 2H).

D147
645.3

¹H NMR (300 MHz, Methanol-d4) δ 8.52 (s, 0.2H, FA), 7.75 (s,

1H), 7.71 (d, J = 5.5 Hz, 1H), 7.61 (d, J = 5.5 Hz, 1H), 7.49 (d, J =

8.2 Hz, 1H), 7.38-7.32 (m, 2H), 7.04 (s, 2H), 5.16 (dd, J = 13.3,

5.2 Hz, 1H), 4.49-4.35 (m, 2H), 4.16 (s, 2H), 3.99 (s, 6H), 3.42

(s, 4H), 3.17 (m, 4H), 2.94-2.85 (m, 1H), 2.85-2.75 (m, 1H),

2.56-2.45 (m, 1H), 2.24-2.13 (m, 1H).

D148
860.5

¹H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.41 (d, J = 8.9 Hz,

1H), 7.95 (d, J = 1.3 Hz, 1H), 7.42 (d, J = 8.5 Hz, 1H), 7.27 (dd, J =

8.5, 2.4 Hz, 1H), 7.17 (d, J = 2.4 Hz, 1H), 6.99 (d, J = 4.5 Hz, 2H),

5.10 (dd, J = 13.2, 5.1 Hz, 1H), 4.43-4.03 (m, 5H), 3.91 (d, J =

3.2 Hz, 6H), 3.82 (d, J = 11.9 Hz, 2H), 3.59 (s, 3H), 3.16 (s, 1H),

2.92 (s, 2H), 2.77 (s, 3H), 2.75-2.56 (m, 11H), 2.46-2.30 (m,

2H), 1.99 (d, J = 12.0 Hz, 1H), 1.80 (s, 4H), 1.56 (d, J = 12.7 Hz,

2H).

D149
767.4

¹H NMR (400 MHz, Methanol-d4) δ 8.56 (s, 1H, FA), 8.27 (s, 1H),

7.85 (s, 1H), 7.45 (d, J = 8.4 Hz, 1H), 7.39-7.30 (m, 2H), 7.05 (s,

2H), 5.15 (dd, J = 13.3, 5.2 Hz, 1H), 4.53 (t, J = 8.3 Hz, 1H), 4.52-

4.34 (m, 2H), 4.31 (s, 2H), 4.01 (s, 6H), 3.75 (s, 3H), 3.31-3.17

(m, 4H), 2.99-2.72 (m, 8H), 2.59-2.43 (m, 1H), 2.46-2.36 (m,

2H), 2.23-2.13 (m, 1H), 2.01-1.97 (m, 2H), 1.88 (t, J = 5.6 Hz,

2H), 1.82 (t, J = 5.6 Hz, 2H).

D150
810.55

¹H NMR (300 MHz, Methanol-d4) δ 8.54 (s, FA 2H), 8.27 (s, 1H),

7.87 (s, 1H), 7.47 (d, J = 8.2 Hz, 1H), 7.35 (d, J = 10.4 Hz, 2H),

7.07 (s, 2H), 5.16 (dd, J = 13.3, 5.2 Hz, 1H), 4.53-4.33 (m, 4H),

4.11-3.97 (m, 7H), 3.96-3.86 (m, 2H), 3.75 (s, 3H), 3.02-2.87

(m, 8H), 2.87-2.65 (m, 6H), 2.58-2.43 (m, 1H), 2.30-2.01 (m,

6H), 1.94-1.69 (m, 4H).

D151
763.35

¹H NMR (300 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.17 (s, 1H), 9.01-

8.91 (m, 1H), 8.59 (d, J = 1.8 Hz, 1H), 8.02 (s, 1H), 7.55 (d, J = 8.4

Hz, 1H), 7.23-7.12 (m, 2H), 7.04 (s, 2H), 5.13-5.01 (m, 1H),

4.62 (s, 1H), 4.41-4.16 (m, 6H), 4.01 (s, 1H), 3.93 (s, 6H), 3.63

(s, 3H), 3.49 (dd, J = 30.5, 13.7 Hz, 1H), 3.22 (t, J = 12.3 Hz, 1H),

2.92 (s, 1H), 2.78 (d, J = 4.6 Hz, 6H), 2.60 (d, J = 17.0 Hz, 1H),

2.40 (d, J = 13.6 Hz, 1H), 2.00 (dd, J = 20.3, 10.8 Hz, 3H).

D152
763.35

¹H NMR (300 MHz, DMSO-d6) δ 11.00 (s, 1H), 9.08 (s, 1H), 9.00-

8.90 (m, 1H), 8.60 (d, J = 2.3 Hz, 1H), 8.02 (s, 1H), 7.47 (d, J = 8.5

Hz, 1H), 7.42-7.32 (m, 1H), 7.28 (d, J = 9.0 Hz, 1H), 7.04 (s, 2H),

5.18-5.05 (m, 1H), 4.60 (s, 1H), 4.36 (d, J = 16.9 Hz, 1H), 4.25

(d, J = 4.4 Hz, 3H), 4.23-4.08 (m, 1H), 3.93 (s, 6H), 3.63 (s, 3H),

3.54-3.07 (m, 3H), 3.02-2.84 (m, 1H), 2.82-2.80 (m, 1H), 2.78

(d, J = 4.6 Hz, 6H), 2.61 (d, J = 16.5 Hz, 1H), 2.46-2.35 (m, 1H),

1.98 (d, J = 25.6 Hz, 3H).

D153
794.2

¹H NMR (400 MHz, Methanol-d4) δ 8.35 (s, 3H, FA), 7.62 (d, J =

5.4 Hz, 1H), 7.47-7.37 (m, 2H), 6.90-6.82 (m, 3H), 6.79 (d, 1H),

5.14 (dd, J = 13.3, 5.1 Hz, 1H), 4.47-4.32 (m, 4H), 3.95 (s, 6H),

3.75 (s, 3H), 3.72 (s, 4H), 3.59 (d, J = 12.0 Hz, 2H), 3.23-3.19

(m, 1H), 3.01-3.75 (m, 6H), 2.68 (s, 3H), 2.57-2.44 (m, 1H),

2.41 (s, 3H), 2.23-2.15 (m, 1H), 2.13-2.01 (m, 7H), 1.63 (s,

2H).

D154
683.3

¹H NMR (400 MHz, DMSO-d6) δ 10.99 (s, 1H), 9.50-9.43 (m, 1H,

TFA), 7.94 (s, 1H), 7.72 (d, J = 5.4 Hz, 1H), 7.67-7.60 (m, 2H),

7.34-7.24 (m, 2H), 7.09 (s, 2H), 5.09 (dd, J = 13.3, 5.1 Hz, 1H),

4.97-4.62 (m, 1H), 4.59-4.49 (m, 1H), 4.40 (d, J = 17.0 Hz, 1H),

4.28 (d, J = 16.9 Hz, 1H), 4.09 (d, J = 13.1 Hz, 1H), 3.98 (s, 6H),

3.83 (d, J = 13.9 Hz, 1H), 3.64 (s, 3H), 3.58 (d, J = 13.5 Hz, 2H),

3.29 (t, J = 11.8 Hz, 1H), 3.07 (d, J = 12.9 Hz, 1H), 2.93 (ddd, J =

17.9, 13.5, 5.4 Hz, 1H), 2.77-2.54 (m, 3H), 2.40 (td, J = 13.1, 4.5

Hz, 1H), 2.14 (s, 3H), 2.03-1.88 (m, 2H).

D155
763.35

¹H NMR (400 MHz, Methanol-d4) δ 8.55 (s, 1H, FA), 8.36 (d, J =

0.7 Hz, 1H), 7.87 (s, 1H), 7.68 (d, J = 9.3 Hz, 1H), 7.20-7.14 (m,

2H), 7.07 (s, 2H), 5.17-5.08 (m, 1H), 4.73-4.63 (m, 1H), 4.50-

4.42 (m, 2H), 4.38 (s, 2H), 4.34-4.22 (m, 1H), 4.09 (d, J = 13.7

Hz, 1H), 4.02 (s, 6H), 3.75 (s, 3H), 3.44-3.38 (m, 1H), 3.23-

3.14 (m, 1H), 2.98-2.92 (m, 1H), 2.89 (s, 6H), 2.83-2.76 (m,

1H), 2.55-2.44 (m, 1H), 2.23-2.13 (m, 2H), 2.13-2.02 (m, 1H).

D156
763.35

¹H NMR (400 MHz, Methanol-d4) δ 8.56 (s, 1H, FA), 8.38 (s, 1H),

7.87 (s, 1H), 7.50 (d, J = 8.3 Hz, 1H), 7.43-7.35 (m, 2H), 7.07 (s,

2H), 5.20-5.13 (m, 1H), 4.71-4.59 (m, 2H), 4.51-4.41 (m, 2H),

4.37 (s, 2H), 4.10 (s, 1H), 4.02 (s, 6H), 3.93 (d, J = 12.9 Hz, 1H),

3.75 (s, 3H), 3.16-3.08 (m, 1H), 2.98-2.91 (m, 1H), 2.87 (s,

6H), 2.84-2.78 (m, 1H), 2.58-2.47 (m, 1H), 2.26-2.15 (m, 2H),

2.13-2.04 (m, 1H).

D157
844.35

¹H NMR (400 MHz, DMSO-d6) δ 8.55 (s, 1H), 7.90 (s, 1H), 7.43

(d, J = 8.4 Hz, 1H), 7.27 (d, J = 8.0 Hz, 1H), 7.17 (s, 2H), 7.11 (s,

1H), 5.05 (dd, J = 13.4, 4.8 Hz, 1H), 4.40-4.15 (m, 3H), 3.86 (s,

4H), 3.82-3.80 (m, 3H), 3.61 (s, 3H), 3.16 (s, 1H), 2.95-2.80 (m,

2H), 2.80-2.65 (m, 4H), 2.64-2.54 (m, 3H), 2.51 (s, 6H), 2.45-

2.35 (m, 2H), 2.04-1.96 (m, 1H), 1.87-1.75 (m, 4H), 1.64-1.47

(m, 2H), 1.17 (t, J = 7.6 Hz, 3H).

D158
696.35

¹H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.92 (s, 1H, TFA

salt), 7.53 (s, 2H), 7.42 (d, J = 8.9 Hz, 1H), 6.84 (s, 2H), 6.72 (s,

2H), 5.08 (dd, J = 13.3, 5.1 Hz, 1H), 4.39-4.17 (m, 4H), 3.88 (s,

6H), 3.77 (s, 2H), 3.65 (s, 2H), 3.62 (s, 3H), 3.21-3.12 (m, 2H),

2.98-2.85 (m, 1H), 2.64-2.60 (m, 2H), 2.57 (s, 1H), 2.42-2.37

(m, 1H), 2.35 (s, 3H), 2.14 (d, J = 14.0 Hz, 2H), 2.00 (t, J = 11.7

Hz, 3H).

D159
849.4

¹H NMR (400 MHz, Methanol-d4) δ 8.57 (s, 1H, FA), 8.39 (d, J =

5.2 Hz, 1H), 7.86 (s, 1H), 7.50 (d, J = 8.4 Hz, 1H), 7.34 (s, 1H),

7.23 (d, J = 8.3, 2.4 Hz, 1H), 7.05 (s, 2H), 5.17 (dd, J = 13.3, 6.9,

5.1 Hz, 1H), 4.49-4.39 (m, 3H), 4.35 (s, 2H), 4.11 (t, 2H), 4.01 (s,

6H), 3.75 (s, 3H), 3.29-3.14 (m, 1H), 3.07-2.98 (m, 1H), 2.97-

2.88 (m, 1H), 2.86 (s, 6H), 2.83-2.76 (m, 1H), 2.59-2.47 (m,

3H), 2.45-2.33 (m, 1H), 2.32-2.25 (m, 1H), 2.24-2.15 (m, 1H),

2.08-1.98 (m, 1H), 1.98-1.92 (m, 1H), 1.92-1.83 (m, 2H), 1.69-

1.52 (m, 4H).

D160
822.4

¹H NMR (400 MHz, Methanol-d4) δ 7.82 (d, J = 8.2 Hz, 1H), 7.67

(s, 1H), 7.30 (d, J = 2.3 Hz, 1H), 7.25 (dd, J = 8.3, 2.3 Hz, 1H),

7.14 (d, J = 1.3 Hz, 1H), 7.04 (d, J = 3.1 Hz, 2H), 5.12 (dd, J =

12.5, 5.4 Hz, 1H), 4.98 (d, J = 6.5 Hz, 1H), 4.40 (s, 2H), 4.00-3.99

(s, 6H), 3.69-3.63 (s, 3H), 3.60-3.49 (m, 4H), 3.37 (s, 1H), 3.23-

3.12 (m, 2H), 3.12-3.07 (m, 2H), 3.07-2.93 (m, 1H), 2.92-

2.74 (m, 4H), 2.66 (d, J = 1.2 Hz, 3H), 2.65 (s, 1H), 2.05 (m, 1H),

2.16-2.05 (m, 6H), 2.03-1.97 (m, 3H), 1.64 (q, J = 12.1 Hz, 2H).

D161
807.43

D162
846.35

¹H NMR (400 MHz, Methanol-d4 with a drop of D2O) δ 8.37 (s,

1H), 8.36 (br s, 1H, FA), 7.87 (s, 1H), 7.45 (d, J = 8.4 Hz, 1H), 7.39-

7.31 (m, 2H), 7.08 (s, 2H), 5.15 (dd, J = 13.3, 5.2 Hz, 1H), 4.49-

4.34 (m, 5H), 4.02 (s, 6H), 3.86 (d, J = 12.3 Hz, 2H), 3.75 (s, 3H),

3.30-3.26 (m, 1H), 3.16-3.09 (m, 1H), 2.92 (s, 7H), 2.86-2.74

(m, 3H), 2.71-2.60 (m, 2H), 2.56-2.48 (m, 2H), 2.24-2.15 (m,

1H), 2.05-1.92 (m, 4H), 1.71 (q, J = 12.6 Hz, 2H).

D163
849.4

¹H NMR (300 MHz, DMSO-d6) δ 10.99 (s, 1H), 8.88 (d, J = 8.8 Hz,

1H), 8.59 (s, 1H), 8.24 (s, 1H, FA), 7.97 (s, 1H), 7.63 (d, J = 8.4

Hz, 1H), 7.17 (d, J = 2.2 Hz, 1H), 7.05 (d, J = 8.4, 2.2 Hz, 1H),

6.93 (s, 2H), 5.08 (dd, J = 13.2, 5.1 Hz, 1H), 4.33 (q, J = 22.5 Hz,

3H), 4.07 (t, J = 6.3 Hz, 2H), 3.85 (s, 6H), 3.62 (s, 5H), 3.20-3.08

(m, 1H), 2.97-2.83 (m, 2H), 2.66-2.56 (m, 1H), 2.46-2.31 (m,

4H), 2.26 (s, 6H), 2.20-2.08 (m, 1H), 2.05-1.92 (m, 1H), 1.91-

1.82 (m, 1H), 1.83-1.71 (m, 3H), 1.57-1.38 (m, 4H).

D164
763.2

¹H NMR (400 MHz, DMSO-d6) δ 10.97 (s, 1H), 8.97-8.90 (m,

1H), 8.57 (d, J = 2.5 Hz, 1H), 8.27 (s, 1H, FA), 7.96 (s, 1H), 7.55

(d, J = 8.4 Hz, 1H), 7.23-7.14 (m, 2H), 6.91 (s, 2H), 5.10-5.02

(m, 1H), 4.75-4.56 (m, 1H), 4.41-4.16 (m, 3H), 4.04-3.92 (m,

1H), 3.84 (s, 6H), 3.51 (s, 3H), 3.57-3.46 (m, 2H), 3.22 (t, J = 12.3

Hz, 1H), 2.97-2.85 (m, 1H), 2.69-2.56 (m, 1H), 2.45-2.32 (m,

1H), 2.17 (s, 6H), 2.06-1.89 (m, 3H).

D165
763.35

¹H NMR (300 MHz, Methanol-d4) δ 8.55 (s, FA, 1H), 8.37 (s, 1H),

7.86 (s, 1H), 7.54-7.45 (m, 1H), 7.43-7.32 (m, 2H), 7.07 (s, 2H),

5.23-5.11 (m, 1H), 4.74-4.56 (m, 1H), 4.48-4.35 (m, 4H), 4.12-

4.02 (m, 7H), 3.97-3.87 (m, 1H), 3.78-3.72 (s, 3H), 3.27-3.05

(m, 2H), 2.88-2.78 (m, 8H), 2.56-2.47 (m, 1H), 2.28-2.14 (m,

2H), 2.13-2.05 (m, 1H).

D166
875.5

¹H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.06 (s, 1H, TFA

salt), 8.91 (dd, J = 9.0, 2.8 Hz, 1H), 8.61 (s, 1H), 8.01 (s, 1H), 7.51

(d, J = 8.4 Hz, 1H), 7.38-7.27 (m, 2H), 7.03 (s, 2H), 5.10 (dd, J =

13.2, 5.1 Hz, 1H), 4.38 (d, J = 17.0 Hz, 2H), 4.28-4.19 (m, 3H),

3.92 (s, 6H), 3.63 (s, 3H), 3.57-3.45 (m, 7H), 3.37-3.35 (m, 4H),

3.05-2.99 (m, 1H), 2.93-2.89 (m, 1H), 2.88-2.83 (m, 2H), 2.80-

2.74 (m, 6H), 2.65-2.58 (m, 2H), 2.43-2.36 (m, 2H), 2.05-

1.96 (m, 1H), 1.94-1.76 (m, 2H).

D167
846.4

¹H NMR (300 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.88 (d, J = 8.7 Hz,

1H), 8.60 (s, 1H), 8.23 (s, 2H, FA salt), 7.97 (s, 1H), 7.43 (d, J =

8.5 Hz, 1H), 7.28 (dd, J = 8.5, 2.3 Hz, 1H), 7.17 (d, J = 2.3 Hz,

1H), 6.93 (s, 2H), 5.10 (dd, J = 13.3, 5.1 Hz, 1H), 4.53-4.09 (m,

4H), 3.86 (s, 6H), 3.80 (s, 2H), 3.64 (d, J = 8.8 Hz, 5H), 3.20-

3.14 (m, 1H), 2.99-2.83 (m, 2H), 2.74 (t, J = 11.9 Hz, 2H), 2.65-

2.54 (m, 3H), 2.44-2.34 (m, 1H), 2.29 (s, 6H), 2.04-1.94 (m,

1H), 1.85-1.79 (m, 4H), 1.62-1.52 (m, 2H).

D168
629.15

¹H NMR (400 MHz, Methanol-d4) δ 7.77 (s, 1H), 7.71 (d, J = 5.4

Hz, 1H), 7.64-7.56 (m, 2H), 7.26 (d, J = 8.5 Hz, 1H), 7.21 (s, 1H),

7.08 (s, 2H), 5.38-5.06 (m, 2H), 4.76 (s, 2H), 4.65 (d, J = 16.6

Hz, 2H), 4.55-4.40 (m, 2H), 4.37-4.32 (m, 2H), 4.08-3.95 (m,

6H), 3.75 (s, 3H), 2.98-2.86 (m, 1H), 2.85-2.76 (m, 1H), 2.58-

2.45 (m, 1H), 2.22-2.18 (m, 1H).

D169
643.1

¹H NMR (400 MHz, Methanol-d4) δ 7.90 (d, J = 8.3 Hz, 1H), 7.77

(s, 1H), 7.71 (d, J = 5.4 Hz, 1H), 7.61 (d, J = 5.4 Hz, 1H), 7.37 (d,

J = 2.2 Hz, 1H), 7.32 (dd, J = 8.3, 2.3 Hz, 1H), 7.08 (s, 2H), 5.34 (s,

1H), 5.15 (dd, J = 12.5, 5.4 Hz, 1H), 4.78 (dd, J = 12.0, 6.3 Hz,

2H), 4.64 (s, 2H), 4.38 (d, J = 10.8 Hz, 2H), 4.00 (s, 6H), 3.75 (s,

3H), 2.96-2.83 (m, 1H), 2.83-2.66 (m, 2H), 2.20-2.10 (m, 1H).

D170
629.1

¹H NMR (400 MHz, Methanol-d4) δ 7.83-7.74 (m, 2H), 7.71 (d,

J = 5.4 Hz, 1H), 7.61 (d, J = 5.4 Hz, 1H), 7.13-7.05 (m, 4H), 5.38-

5.08 (m, 2H), 4.77 (s, 2H), 4.66 (s, 2H), 4.55-4.40 (m, 2H), 4.35

(s, 2H), 4.07-3.92 (m, 6H), 3.75 (s, 3H), 3.00-2.86 (m, 1H),

2.85-2.75 (m, 1H), 2.57-2.42 (m, 1H), 2.27-2.10 (m, 1H).

D171
889.45

¹H NMR (300 MHz, DMSO-d6) δ 11.00 (s, 1H), 9.08 (s, 1H, TFA

salt), 8.83 (d, J = 8.7 Hz, 1H), 8.53 (s, 1H), 7.52 (d, J = 8.3 Hz,

1H), 7.40-7.28 (m, 2H), 6.82 (d, J = 2.2 Hz, 2H), 5.11 (dd, J =

13.2, 5.0 Hz, 1H), 4.42-4.22 (m, 5H), 3.86 (s, 6H), 3.62 (s, 3H),

3.50-3.45 (m, 8H), 3.07-2.95 (m, 2H), 2.95-2.90 (m, 1H), 2.89-

2.74 (m, 9H), 2.68-2.54 (m, 2H), 2.45-2.30 (m, 6H), 2.01-

1.79 (m, 3H).

D172
941.40

¹H NMR (300 MHz, MeOD) δ 8.27 (s, 1H), 7.42 (d, J = 8.2 Hz, 1H),

6.92-6.76 (m, 4H), 5.14 (dd, J = 13.2, 5.1 Hz, 1H), 4.85-4.68 (m,

1H), 4.67-4.51 (m, 2H), 4.48-4.31 (m, 4H), 4.23-4.18 (m, 2H),

3.96 (s, 7H), 3.81-3.70 (m, 7H), 3.69-3.39 (m, 8H), 3.26-3.05

(m, 2H), 3.02-2.72 (m, 5H), 2.60-2.40 (m, 4H), 2.36-2.08 (m,

7H).

D173
822.35

¹H NMR (300 MHz, Methanol-d4) δ 8.39 (s, 1H, FA), 7.82 (d, J =

8.3 Hz, 1H), 7.71 (s, 1H), 7.36 (d, J = 1.4 Hz, 1H), 7.33-7.22 (m,

2H), 7.04 (s, 2H), 5.13 (dd, J = 12.4, 5.4 Hz, 1H), 5.03-4.96 (m,

1H), 4.69-4.59 (m, 1H), 4.40 (s, 2H), 4.01 (s, 6H), 3.74 (s, 3H),

3.63-3.52 (m, 2H), 3.22-3.13 (m, 2H), 3.11-2.96 (m, 3H), 2.93-

2.70 (m, 5H), 2.64-2.56 (m, 5H), 2.23-2.04 (m, 6H), 2.02-

1.87 (m, 4H), 1.91-1.53 (m, 2H).

D174
794.25

¹H NMR (400 MHz, DMSO-d6) δ 7.82 (s, 1H), 7.69-7.59 (m, 2H),

7.51 (d, J = 8.3 Hz, 1H), 7.16-7.07 (m, 2H), 6.99 (s, 2H), 4.99

(dd, J = 13.3, 5.1 Hz, 1H), 4.86-4.78 (m, 1H), 4.42-4.24 (m,

2H), 4.21 (d, J = 6.4 Hz, 2H), 3.88 (s, 6H), 3.60 (s, 3H), 3.37 (dd,

J = 28.0, 12.1 Hz, 4H), 3.16 (s, 1H), 3.01 (t, J = 12.7 Hz, 2H), 2.92

(d, J = 6.9 Hz, 2H), 2.90-2.78 (m, 2H), 2.70-2.64 (m, 1H), 2.39

(s, 2H), 2.08-1.73 (m, 11H), 1.44 (q, J = 12.1 Hz, 2H).

D175
915.35

¹H NMR (300 MHz, DMSO-d6) δ 10.99 (s, 1H), 9.07 (s, 1H, TFA

salt), 8.93 (d, J = 8.8 Hz, 1H), 8.62 (s, 1H), 8.02 (s, 1H), 7.42 (d,

J = 8.9 Hz, 1H), 7.03 (s, 2H), 6.77-6.65 (m, 2H), 5.08 (dd, J = 13.2,

5.1 Hz, 1H), 4.49-4.38 (m, 1H), 4.39-4.14 (m, 5H), 3.93 (s, 6H),

3.70 (s, 2H), 3.64 (s, 3H), 3.29 (s, 6H), 3.07-2.95 (m, 2H), 2.95-

2.89 (m, 1H), 2.91-2.69 (m, 9H), 2.68-2.56 (m, 2H), 2.45-2.28

(m, 2H), 2.17-1.78 (m, 7H).

D176
682.45

¹H NMR (400 MHz, DMSO-d6) δ 11.00 (s, 1H), 9.32 (d, J = 7.7 Hz,

1H, TFA), 7.94 (s, 1H), 7.72 (d, J = 5.4 Hz, 1H), 7.63 (d, J = 5.4

Hz, 1H), 7.55 (d, J = 8.4 Hz, 1H), 7.49-7.39 (m, 2H), 7.09 (s, 2H),

5.14 (dd, J = 13.3, 5.1 Hz, 1H), 4.53 (dd, J = 13.2, 7.9 Hz, 1H),

4.40 (d, J = 16.9 Hz, 1H), 4.27 (dd, J = 17.3, 1.8 Hz, 1H), 4.09 (d,

J = 13.1 Hz, 1H), 3.98 (s, 6H), 3.88 (d, J = 13.7 Hz, 1H), 3.73 (s,

2H), 3.64 (s, 2H), 3.51 (s, 1H), 3.47 (d, J = 7.4 Hz, 1H), 3.31 (s,

1H), 3.08 (d, J = 12.8 Hz, 1H), 3.00-2.87 (m, 1H), 2.61 (d, J =

19.7 Hz, 3H), 2.40 (dt, J = 13.3, 6.7 Hz, 1H), 2.22 (d, J = 10.8 Hz,

2H), 2.12 (q, J = 8.9, 8.5 Hz, 1H), 2.06-1.98 (m, 1H), 1.92 (d, J =

11.0 Hz, 1H).

D177
749.25

¹H NMR (400 MHz, Methanol-d4) δ 8.57 (s, 1H, FA), 8.39 (s, 1H),

7.86 (s, 1H), 7.49 (d, J = 8.2 Hz, 1H), 7.05 (s, 3H), 7.03-6.97 (m,

1H), 5.24-5.12 (m, 2H), 4.48-4.35 (m, 2H), 4.21 (s, 2H), 4.06-

3.97 (m, 7H), 3.96-3.83 (m, 2H), 3.75 (s, 3H), 3.57 (t, J = 8.4 Hz,

1H), 2.99-2.87 (m, 1H), 2.85-2.80 (m, 1H), 2.74 (s, 6H), 2.56-

2.46 (m, 1H), 2.24-2.15 (m, 1H).

D178
682.2

¹H NMR (400 MHz, DMSO-d6) δ 10.94 (s, 1H), 8.28 (s, 1H), 7.87

(s, 1H), 7.69 (d, J = 5.4 Hz, 1H), 7.60 (d, J = 5.3 Hz, 1H), 7.47 (d,

J = 8.3 Hz, 1H), 6.92 (s, 2H), 6.54-6.44 (m, 2H), 5.03 (dd, J = 13.3,

5.1 Hz, 1H), 4.30 (d, J = 16.9 Hz, 1H), 4.17 (d, J = 16.8 Hz, 1H),

3.85 (s, 6H), 3.61 (s, 6H), 3.52 (s, 2H), 2.98-2.83 (m, 1H), 2.58

(d, J = 17.1 Hz, 1H), 2.46-2.28 (m, 6H), 2.02-1.88 (m, 1H), 1.72

(t, J = 5.3 Hz, 4H).

D179
848.25

¹H NMR (400 MHz, DMSO-d6) δ 11.06 (s, 1H), 9.01 (s, 1H), 8.95

(s, 1H, TFA), 8.61 (s, 1H), 8.02 (s, 1H), 7.73 (d, J = 7.8 Hz, 1H),

7.03 (s, 2H), 6.71 (d, J = 7.9 Hz, 1H), 6.17 (s, 1H), 5.24 (dd, J =

12.4, 5.2 Hz, 1H), 4.25 (d, J = 5.0 Hz, 2H), 3.93 (s, 9H), 3.64 (s,

3H), 2.92-2.83 (m, 4H), 2.81-2.75 (m, 7H), 2.68-2.59 (m, 2H),

2.51-2.41 (m, 2H), 2.17-2.04 (m, 2H), 1.98-1.82 (m, 4H), 1.64-

1.47 (m, 2H).

D180
749.3

¹H NMR (300 MHz, Methanol-d4) δ 8.38 (s, 1H), 7.88 (s, 1H), 7.69

(d, J = 8.8 Hz, 1H), 7.08 (s, 2H), 6.88-6.68 (m, 2H), 5.32-5.07

(m, 2H), 4.54-4.3 (m, 4H), 4.03 (s, 6H), 3.99-3.88 (m, 2H), 3.75

(s, 3H), 3.67-3.54 (m, 1H), 2.92 (s, 7H), 2.83-2.72 (m, 2H), 2.58-

2.38 (m, 1H), 2.23-2.09 (m, 1H).

D181
832.35

¹H NMR (400 MHz, Methanol-d4) δ 8.56 (s, 1H, FA), 8.32 (s, 1H),

7.86 (s, 1H), 7.65 (d, 1H), 7.11 (d, J = 7.6 Hz, 2H), 7.05 (s, 2H),

5.12 (dd, J = 13.3, 5.1 Hz, 1H), 4.83-4.77 (m, 1H), 4.48-4.35 (d,

J = 16.8 Hz, 2H), 4.32 (s, 2H), 4.01 (s, 6H), 3.93 (d, J = 13.0 Hz,

2H), 3.75 (s, 3H), 3.45-3.38 (m, 2H), 3.08-2.95 (m, 3H), 2.95-

2.89 (m, 1H), 2.84 (s, 6H), 2.81-2.70 (m, 2H), 2.57-2.42 (m,

2H), 2.21-2.13 (m, 1H), 2.04 (t, J = 12.7 Hz, 2H), 1.74-1.56 (m,

2H).

D182
832.35

¹H NMR (400 MHz, Methanol-d4) δ 8.65 (br s, FA, 1H), 8.33 (s,

1H), 7.86 (s, 1H), 7.46 (d, J = 8.3 Hz, 1H), 7.39-7.30 (m, 2H),

7.06 (s, 2H), 5.15 (dd, J = 13.4, 5.1 Hz, 1H), 4.83-4.78(m, 1H),

4.51-4.39 (m, 2H), 4.33 (s, 2H), 4.01 (s, 6H), 3.87-3.68 (m,

5H), 3.48-3.39 (m, 2H), 3.08-2.96 (m, 1H), 2.97-2.88 (m, 3H),

2.85 (s, 6H), 2.83-2.71 (m, 2H), 2.58-2.39 (m, 2H), 2.23-2.15

(m, 1H), 2.13-1.98 (m, 2H), 1.79-1.59(m, 2H).

D183
696.15

¹H NMR (400 MHz, DMSO-d6) δ 11.08 (s, 1H), 8.21 (s, 1H, FA),

7.87 (s, 1H), 7.69 (d, J = 5.4 Hz, 1H), 7.66-7.55 (m, 2H), 6.93 (s,

2H), 6.78 (d, J = 2.1 Hz, 1H), 6.69-6.60 (m, 1H), 5.12-4.97 (m,

1H), 3.85 (s, 6H), 3.73 (s, 4H), 3.62 (s, 3H), 3.53 (s, 2H), 2.97-

2.81 (m, 1H), 2.70-2.53 (m, 2H), 2.46-2.31 (m, 4H), 2.07-1.95

(m, 1H), 1.80-1.67 (m, 4H).

D184
807.35

¹H NMR (300 MHz, DMSO-d6) δ 11.09 (s, 1H), 9.45 (s, 1H, TFA

salt), 9.23 (s, 1H, TFA salt), 7.85 (s, 1H), 7.69 (dd, J = 8.3, 2.6 Hz,

1H), 7.33 (d, J = 1.4 Hz, 1H), 7.03 (s, 2H), 6.78 (d, J = 2.2 Hz, 1H),

6.66 (dd, J = 8.3, 2.1 Hz, 1H), 5.06 (dd, J = 12.9, 5.4 Hz, 1H), 4.35-

4.19 (m, 2H), 3.99-3.79 (m, 10H), 3.61 (s, 3H), 3.54-3.50 (m,

2H), 3.48-3.46 (m, 2H), 3.25-3.16 (m, 2H), 3.10-2.83 (m, 7H),

2.66-2.60 (m, 1H), 2.56-2.53 (m, 3H), 2.20-2.10 (m, 2H), 2.06-

1.88 (m, 5H), 1.58-1.39 (m, 2H).

D185
793.25

¹H NMR (300 MHz, Methanol-d4) δ 8.49 (s, 1H, FA), 7.80-7.57

(m, 4H), 7.07 (s, 2H), 6.83 (d, J = 2.0 Hz, 1H), 6.67 (dd, J = 8.4,

2.1 Hz, 1H), 5.07 (dd, J = 12.4, 5.4 Hz, 1H), 4.39 (s, 2H), 4.01 (s,

6H), 3.79 (d, J = 16.3 Hz, 7H), 3.55 (d, J = 11.6 Hz, 2H), 3.23-

3.07 (m, 2H), 2.96-2.81 (m, 1H), 2.81-2.67 (m, 2H), 2.58 (s,

4H), 2.39 (s, 2H), 2.16-2.00 (m, 3H), 1.99-1.89 (m, 5H), 1.55

(s, 2H).

D186
682.2

¹H NMR (400 MHz, DMSO-d6) δ 7.82 (s, 1H), 7.73-7.55 (m, 2H),

7.40 (d, J = 8.9 Hz, 1H), 6.98 (s, 2H), 6.83-6.67 (m, 2H), 5.09-

4.91 (m, 1H), 4.44-4.12 (m, 4H), 3.89 (s, 6H), 3.73 (s, 2H), 3.61

(s, 5H), 3.34 (d, J = 12.2 Hz, 2H), 3.09 (t, J = 11.9 Hz, 2H), 2.90-

2.75 (m, 1H), 2.70-2.57 (m, 1H), 2.44-2.28 (m, 1H), 2.17-1.88

(m, 5H).

D187
846.1

¹H NMR (400 MHz, DMSO-d6) δ 11.09 (s, 1H), 8.97 (d, J = 8.4 Hz,

1H), 8.53 (s, 1H), 8.18 (s, 1H, FA), 7.98 (s, 1H), 7.68 (d, J = 8.5

Hz, 1H), 7.36 (s, 1H), 7.28 (dd, J = 8.7, 2.3 Hz, 1H), 6.92 (s, 2H),

5.08 (dd, J = 12.9, 5.4 Hz, 1H), 4.71-4.58 (m, 1H), 4.01 (d, J =

12.9 Hz, 2H), 3.85 (s, 6H), 3.62 (s, 3H), 3.56 (s, 2H), 3.08 (t, J =

11.7 Hz, 3H), 2.96-2.76(m, 3H), 2.73-2.53 (m, 4H), 2.22 (s,

6H), 2.08-1.86 (m, 3H), 1.53-1.46 (m, 2H).

D188
927.58

¹H NMR (400 MHz, Methanol-d4) δ 8.55 (s, 2H, FA), 8.36 (s, 1H),

7.85 (s, 1H), 7.39 (d, J = 8.2 Hz, 1H), 7.06 (s, 2H), 6.85 (d, J = 2.1

Hz, 1H), 6.77 (dd, J = 8.2, 2.2 Hz, 1H), 5.14 (dd, J = 13.2, 5.1 Hz,

1H), 4.62 (s, 1H), 4.47 (s, 2H), 4.46-4.32 (m, 2H), 4.22 (t, J = 9.6

Hz, 2H), 4.02 (s, 6H), 3.95 (t, J = 9.3 Hz, 2H), 3.75 (s, 3H), 3.67 (s,

4H), 3.23-3.04 (m, 2H), 2.99-2.89 (m, 2H), 2.85-2.74(m, 1H),

2.69 (d, J = 6.9 Hz, 2H), 2.55-2.45 (m, 5H), 2.39 (s, 3H), 2.29 (t,

J = 11.7 Hz, 1H), 2.21-2.13 (m, 1H), 2.08-1.99 (m, 1H), 1.98-

1.86 (m, 4H), 1.31 (s, 2H).

D189
808.35

¹H NMR (300 MHz, DMSO-d6) δ 11.14 (s, 1H), 9.22 (s, 2H, TFA),

7.94-7.83 (m, 2H), 7.75-7.60 (m, 2H), 7.37-7.26 (m, 2H), 7.06

(d, J = 1.7 Hz, 2H), 5.17-4.97 (m, 2H), 4.27 (d, J = 27.8 Hz, 2H),

3.94 (s, 6H), 3.63 (s, 3H), 3.55-3.49 (m, 1H), 3.46 (s, 3H), 3.26-

3.17 (m, 2H), 3.06-2.83 (m, 6H), 2.68-2.53 (m, 3H), 2.10-1.79

(m, 10H), 1.58-1.39 (m, 2H).

D190
763.15

¹H NMR (300 MHz, DMSO-d6) δ 11.11 (s, 1H), 9.25 (d, J = 8.5 Hz,

1H), 8.55 (s, 1H), 8.17 (s, 1H, FA), 7.99 (s, 1H), 7.76 (d, J = 8.4

Hz, 1H), 7.09 (d, J = 2.2 Hz, 1H), 6.97 (dd, J = 8.5, 2.2 Hz, 1H),

6.93 (s, 2H), 5.16-5.04 (m, 2H), 4.14-4.01 (m, 3H), 3.86 (s,

6H), 3.66-3.57 (m, 6H), 3.00-2.81 (m, 1H), 2.65-2.54 (m, 2H),

2.25 (s, 6H), 2.07-2.01 (m, 1H).

D191
832.4

¹H NMR (300 MHz, Methanol-d4) δ 8.56 (s, FA, 1H), 8.34 (s, 1H),

7.84 (s, 1H), 7.62 (d, J = 8.3 Hz, 1H), 7.02 (s, 2H), 6.82 (d, J = 2.1

Hz, 1H), 6.65 (dd, J = 8.3, 2.1 Hz, 1H), 5.07 (dd, J = 12.4, 5.4 Hz,

1H), 4.60-4.39 (m, 1H), 4.27 (s, 2H), 4.15 (t, J = 7.8 Hz, 2H), 4.00

(s, 6H), 3.96-3.83 (m, 2H), 3.72 (s, 3H), 3.63-3.51 (m, 1H), 3.28-

3.21 (m, 1H), 3.09-3.00 (m, 1H), 2.95-2.60 (m, 9H), 2.58-

2.39 (m, 1H), 2.36-2.23 (m, 1H), 2.18-1.93 (m, 3H).

D192
777.25

¹H NMR (300 MHz, DMSO-d6) δ 11.12 (s, 1H), 9.05 (s, 1H, TFA),

8.94 (d, J = 8.8 Hz, 1H), 8.57 (s, 1H), 8.02 (s, 1H), 7.72 (d, J = 8.5

Hz, 1H), 7.51 (s, 1H), 7.41 (d, J = 8.6 Hz, 1H), 7.04 (s, 2H), 5.10

(dd, J = 12.8, 5.3 Hz, 1H), 4.76-4.63 (m, 1H), 4.58-4.52 (m,

1H), 4.25 (d, J = 4.8 Hz, 2H), 4.18 (d, J = 13.6 Hz, 1H), 3.93 (s,

6H), 3.76-3.51 (m, 5H), 2.93-2.83 (m, 1H), 2.82-2.75 (m, 6H),

2.66-2.54 (m, 2H), 2.09-1.92 (m, 3H).

D193
794.2

¹H NMR (300 MHz, DMSO-d6) δ 8.28 (s, 3H), 8.08 (d, J = 5.4 Hz,

1H), 7.76 (s, 1H), 7.62 (d, J = 8.4 Hz, 1H), 7.50 (d, J = 5.1 Hz, 1H),

7.03 (s, 1H), 6.96 (d, J = 8.4 Hz, 1H), 6.82 (s, 2H), 5.02 (dd, J =

13.2, 5.1 Hz, 1H), 4.85-4.74 (m, 1H), 4.38 (d, J = 17.1 Hz, 1H),

4.24 (d, J = 17.1 Hz, 1H), 3.96 (s, 2H), 3.86 (s, 6H), 3.62 (s, 3H),

3.16 (d, J = 9 Hz, 2H), 2.96-2.80 (m, 1H), 2.74-2.52 (m, 3H),

2.48-2.12 (m, 9H), 2.05-1.90 (m, 1H), 1.85-1.50 (m, 9H), 1.40-

1.20 (m, 2H).

D194
642.15

¹H NMR (400 MHz, DMSO-d6) δ 10.97 (s, 1H), 8.15 (s, 1H), 7.88

(s, 1H), 7.69 (d, J = 5.4 Hz, 1H), 7.60 (d, J = 5.3 Hz, 1H), 7.42 (d,

J = 8.5 Hz, 1H), 7.25 (dd, J = 8.5, 2.4 Hz, 1H), 7.14 (d, J = 2.3 Hz,

1H), 6.94 (s, 2H), 5.09 (dd, J = 13.3, 5.1 Hz, 1H), 4.37-4.14 (m,

2H), 3.87 (s, 6H), 3.62 (d, J = 3.5 Hz, 5H), 3.17 (t, J = 4.9 Hz, 4H),

2.91 (ddd, J = 17.2, 13.6, 5.4 Hz, 1H), 2.61 (d, J = 5.0 Hz, 5H),

2.45-2.33 (m, 1H), 2.03-1.95 (m, 1H).

D195
874.2

¹H NMR (300 MHz, DMSO-d6) δ 11.00 (s, 1H), 8.89 (d, J = 9.0 Hz,

1H), 8.59 (s, 1H), 8.26 (s, 3H, FA), 7.96 (s, 1H), 7.65 (d, J = 7.8

Hz, 1H), 7.50 (s, 1H), 7.41 (d, J = 8.0 Hz, 1H), 6.92 (s, 2H), 5.11

(dd, J = 13.2, 5.1 Hz, 1H), 4.43-4.29 (m, 2H), 3.84 (s, 6H), 3.65-

3.59 (m, 5H), 3.24-3.00 (m, 3H), 2.98-2.79 (m, 3H), 2.66-2.55

(m, 7H), 2.21 (s, 6H), 2.22-2.04 (m, 3H), 2.04-1.88 (m, 2H),

1.79-1.76 (m, 6H).

D196
875.35

¹H NMR (400 MHz, Methanol-d4) δ 8.91 (dd, J = 8.8 Hz, 0.16H,

TFA salt), 8.34 (s, 1H), 7.88 (s, 1H), 7.73 (d, J = 9.0 Hz, 1H), 7.20

(d, J = 7.7 Hz, 2H), 7.07 (s, 2H), 5.14 (dd, J = 13.3, 5.1 Hz, 1H),

4.51 (s, 3H), 4.43 (d, J = 15.7 Hz, 2H), 4.02 (s, 6H), 3.75 (s, 3H),

3.68 (d, J = 11.2 Hz, 4H), 3.62 (s, 4H), 3.49-3.43 (m, 2H), 3.38

(s, 2H), 3.17 (d, J = 11.5 Hz, 1H), 2.99 (t, J = 5.3 Hz, 2H), 2.92 (s,

6H), 2.80 (d, J = 17.3 Hz, 1H), 2.65 (dd, J = 28.9, 12.3 Hz, 1H),

2.57-2.41 (m, 2H), 2.22-2.14 (m, 1H), 2.12-2.04 (m, 1H), 2.02-

1.97 (m, 1H).

D197
735.34

¹H NMR (400 MHz, Methanol-d4) δ 8.00 (s, 1H), 7.86 (d, J = 2.1

Hz, 1H), 7.41 (d, J = 8.2 Hz, 1H), 7.16 (s, 2H), 7.09 (d, J = 2.1 Hz,

1H), 6.87 (d, J = 2.1 Hz, 1H), 6.79 (dd, J = 8.2, 2.1 Hz, 1H), 5.13

(dd, J = 13.3, 5.1 Hz, 1H), 4.54 (d, J = 31.5 Hz, 2H), 4.37 (dd, J =

16.3, 9.0 Hz, 4H), 4.17 (d, J = 23.2 Hz, 2H), 4.02 (s, 6H), 3.76 (s,

7H), 3.54 (d, J = 12.4 Hz, 4H), 3.18-2.98 (m, 2H), 2.91 (s, 4H),

2.79 (ddd, J = 17.6, 4.7, 2.4 Hz, 1H), 2.50 (qd, J = 13.2, 4.7 Hz,

1H), 2.34-2.02 (m, 4H).

D198
736.2

¹H NMR (300 MHz, DMSO-d6) δ 8.74 (s, 1H), 8.35 (s, 1H), 7.40 (s,

1H), 7.14 (s, 2H), 6.70 (s, 2H), 5.06 (dd, J = 13.1, 5.1 Hz, 1H),

4.45-4.27 (m, 3H), 4.19 (d, J = 16.7 Hz, 3H), 4.02 (d, J = 10.1

Hz, 2H), 3.95 (s, 6H), 3.88 (s, 1H), 3.72 (s, 2H), 3.67 (s, 3H), 3.65

(s, 2H), 3.43 (s, 2H), 3.16 (s, 1H), 3.05-2.81 (m, 3H), 2.76-2.54

(m, 1H), 2.28 (s, 2H), 2.12 (d, J = 13.6 Hz, 2H), 2.02-1.80 (m,

3H).

D199
794.2

¹H NMR (300 MHz, DMSO-d6) δ 8.28 (s, 3H), 8.08 (d, J = 5.4 Hz,

1H), 7.76 (s, 1H), 7.62 (d, J = 8.4 Hz, 1H), 7.50 (d, J = 5.1 Hz, 1H),

7.03 (s, 1H), 6.96 (d, J = 8.4 Hz, 1H), 6.82 (s, 2H), 5.02 (dd, J =

13.2, 5.1 Hz, 1H), 4.85-4.74 (m, 1H), 4.38 (d, J = 17.1 Hz, 1H),

4.24 (d, J = 17.1 Hz, 1H), 3.96 (s, 2H), 3.86 (s, 6H), 3.62 (s, 3H),

3.16 (d, J = 9 Hz, 2H), 2.96-2.80 (m, 1H), 2.74-2.52 (m, 3H),

2.48-2.12 (m, 9H), 2.05-1.90 (m, 1H), 1.85-1.50 (m, 9H), 1.40-

1.20 (m, 2H).

D200
875.45

¹H NMR (400 MHz, Methanol-d4) δ 8.48-8.35 (m, 1H), 7.87 (s,

1H), 7.54 (d, J = 8.8 Hz, 1H), 7.46-7.34 (m, 2H), 7.07 (s, 2H),

5.18 (ddd, J = 20.4, 13.3, 5.1 Hz, 1H), 4.55-4.44 (m, 5H), 4.47-

4.36 (m, 6H), 4.02 (s, 3H), 3.70-3.54 (m, 6H), 3.65-3.60 (m,

2H), 3.49-3.42 (m, 1H), 3.15-3.08 (m, 2H), 3.01-2.94 (m, 2H),

2.92 (s, 6H), 2.88-2.86 (m, 3H), 2.85-2.82 (m, 1H), 2.83-2.73

(m, 1H), 2.59-2.52 (m, 1H), 2.51-2.37 (m, 1H), 2.24-2.15 (m,

1H), 2.00-1.95 (m, 1H).

D201
752.35

¹H NMR (400 MHz, DMSO-d6) δ 10.97 (s, 1H), 9.18 (s, 1H), 7.93

(s, 1H), 7.42-7.36 (m, 1H), 6.95 (s, 2H), 6.68 (d, J = 7.2 Hz, 2H),

5.08 (dd, J = 13.3, 5.1 Hz, 1H), 4.29 (s, 1H), 4.17 (d, J = 24.7 Hz,

3H), 3.93 (s, 8H), 3.68 (s, 3H), 3.58 (s, 6H), 2.98-2.76 (m, 2H),

2.61 (s, 1H), 2.56 (d, J = 7.4 Hz, 2H), 2.45-2.29 (m, 5H), 2.05-

1.93 (m, 1H), 1.79-1.71 (m, 4H).

D202
697.3
. ¹H NMR (400 MHz, DMSO-d6) δ 11.08 (s, 1H), 9.24 (s, 1H), 8.21

(s, 1H, FA), 8.06 (s, 1H), 7.63 (d, J = 8.3 Hz, 1H), 6.91 (s, 2H),

6.78 (s, 1H), 6.65 (d, J = 8.4, 2.1 Hz, 1H), 5.05 (dd, J = 12.9, 5.4

Hz, 1H), 3.85 (s, 7H), 3.73 (s, 4H), 3.66 (s, 4H), 3.52 (s, 2H), 2.95-

2.81 (m, 1H), 2.60 (s, 1H), 2.40 (s, 3H), 2.05-1.95 (m, 1H),

1.73 (s, 4H).

D203
683.5

¹H NMR (300 MHz, Methanol-d4) δ 9.04 (s, 1H), 7.79 (s, 1H), 7.42

(d, J = 8.2 Hz, 1H), 7.10-6.94 (m, 2H), 6.89 (d, J = 2.2 Hz, 1H),

6.85-6.75 (m, 1H), 5.15 (dd, J = 13.3, 5.1 Hz, 1H), 4.57-4.26

(m, 4H), 4.11-3.92 (m, 7H), 3.78 (s, 7H), 3.56 (s, 2H), 3.09 (s,

1H), 3.05-2.72 (m, 2H), 2.68-2.38 (m, 1H), 2.37-1.94 (m, 5H).

D204
683.2

¹H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.28 (s, 1H), 8.47

(s, 1H, FA), 8.10 (s, 1H), 7.29 (d, J = 8.2 Hz, 1H), 7.02 (s, 2H),

6.96-6.87 (m, 2H), 6.20 (s, 1H, FA salt), 5.08 (dd, J = 13.3, 5.1

Hz, 1H), 4.51 (s, 2H), 4.38-4.09 (m, 2H), 3.86 (s, 6H), 3.68 (s,

3H), 3.63 (s, 3H), 3.51 (s, 2H), 3.21-3.13 (m, 2H), 2.98-2.87

(m, 1H), 2.65-2.56 (m, 2H), 2.47-2.35 (m, 1H), 2.14 (s, 2H),

2.01-1.91 (m, 1H), 1.79 (s, 2H).

D205
666.45

¹H NMR (400 MHz, DMSO-d6) δ 10.97 (s, 1H), 7.42 (d, J = 8.4 Hz,

1H), 7.31 (d, J = 1.2 Hz, 1H), 7.25 (dd, J = 8.5, 2.4 Hz, 1H), 7.14

(d, J = 2.3 Hz, 1H), 6.59-6.52 (m, 2H), 5.09 (dd, J = 13.3, 5.1 Hz,

1H), 4.39-4.16 (m, 2H), 3.81 (s, 6H), 3.62 (s, 2H), 3.54 (s, 3H),

3.22-3.11 (m, 4H), 2.98-2.84 (m, 1H), 2.72-2.56 (m, 5H), 2.46-

2.36 (m, 1H), 2.33 (s, 3H), 2.04 (s, 3H), 2.02-1.94 (m, 1H).

D206
630.2

¹H NMR (400 MHz, DMSO-d6) δ 10.99 (s, 1H), 9.25 (s, 1H), 8.14

(s, 0.1H, FA), 8.09 (s, 1H), 7.67 (d, J = 8.3 Hz, 1H), 7.09 (s, 1H),

7.05-6.97 (m, 3H), 6.54 (s, 0.2H, FA salt), 5.09 (dd, J = 13.2, 5.1

Hz, 2H), 4.47-4.15 (m, 6H), 3.90 (s, 8H), 3.67 (s, 3H), 3.03-

2.77 (m, 1H), 2.71-2.60 (m, 1H), 2.43-2.30 (m, 1H), 2.10-1.93

(m, 1H).

D207
682.3

¹H NMR (300 MHz, DMSO-d6) δ 10.97 (s, 1H), 8.22 (s, 1H, FA),

8.09 (d, J = 5.3 Hz, 1H), 7.78 (s, 1H), 7.52 (d, J = 5.3 Hz, 1H), 7.37

(d, J = 8.0 Hz, 1H), 6.78 (s, 2H), 6.69 (d, J = 7.9 Hz, 2H), 5.08 (dd,

J = 13.3, 5.1 Hz, 1H), 4.37-4.11 (m, 2H), 3.84 (s, 6H), 3.63 (s,

3H), 3.55 (d, J = 10.1 Hz, 6H), 2.98-2.82 (m, 1H), 2.62 (s, 1H),

2.45-2.21 (s, 5H), 2.06-1.90 (m, 1H), 1.72 (t, J = 5.5 Hz, 4H).

D208
696.25

¹H NMR (300 MHz, DMSO-d6) δ 11.07 (s, 1H), 8.55 (d, J = 3.1 Hz,

1H), 8.16 (FA, s, 1H), 7.89 (d, J = 3.2 Hz, 1H), 7.63 (d, J = 8.4 Hz,

1H), 7.32 (s, 1H), 6.81 (d, J = 16.7 Hz, 3H), 6.72-6.57 (m, 1H),

5.17-4.96 (m, 1H), 3.85 (s, 6H), 3.81-3.68 (m, 5H), 3.57 (s,

3H), 3.49 (s, 4H), 2.98-2.79 (m, 2H), 2.76-2.63 (m, 1H), 2.63-

2.55 (m, 1H), 2.07-1.90 (m, 1H), 1.82-1.65 (m, 4H).

D209
697.3

¹H NMR (300 MHz, Methanol-d4) δ 9.10 (s, 1H), 8.01 (s, 1H), 7.66

(d, J = 8.3 Hz, 1H), 7.03 (s, 2H), 6.86 (d, J = 2.1 Hz, 1H), 6.70 (dd,

J = 8.3, 2.1 Hz, 1H), 5.08 (dd, J = 12.4, 5.4 Hz, 1H), 4.21 (s, 2H),

4.01 (s, 6H), 3.88 (s, 4H), 3.77 (s, 3H), 3.28-3.00 (m, 4H), 2.97-

2.58 (m, 3H), 2.22-1.99 (m, 5H).

D210
667.35

¹H NMR (400 MHz, DMSO-d6) δ 10.96 (s, 1H), 8.74 (s, 1H), 8.30

(s, 1H), 8.14 (s, 1H, FA), 7.37 (d, J = 8.0 Hz, 1H), 7.06 (s, 2H),

6.70-6.69 (m, 2H), 5.08 (dd, J = 13.2, 5.1 Hz, 1H), 4.33-4.16 (m,

2H), 3.88 (s, 6H), 3.72-3.65 (m, 5H), 3.58 (s, 4H), 2.98-2.82 (m,

1H), 2.66-2.56 (m, 4H), 2.45-2.24 (m, 2H), 2.05-1.96 (m, 1H),

1.78 (s, 4H).

D211
662.3

¹H NMR (300 MHz, MeOD) δ 8.17-8.06 (m, 3H), 7.77-7.68 (m,

3H), 7.57 (d, J = 7.1 Hz, 2H), 7.51 (d, J = 8.6 Hz, 1H), 5.17 (dd, J =

13.3, 5.2 Hz, 1H), 4.56-4.37 (m, 2H), 4.25-4.12 (m, 1H), 3.85

(d, J = 12.7 Hz, 2H), 3.75 (s, 3H), 3.26-3.19 (m, 2H), 3.03-2.74

(m, 2H), 2.62-2.41 (m, 1H), 2.25-2.13 (m, 3H), 2.03-1.85 (m,

2H).

D212
630.3

¹H NMR (300 MHz, DMSO-d6) δ 10.99 (s, 1H), 9.23 (s, 1H), 8.24

(s, 1H, FA), 8.06 (s, 1H), 7.50 (d, J = 8.4 Hz, 1H), 7.12 (dd, J =

8.2, 2.5 Hz, 1H), 7.02 (d, J = 2.4 Hz, 1H), 6.91 (s, 2H), 5.09 (dd,

J = 13.1, 5.0 Hz, 1H), 4.85-4.75 (m, 1H), 4.45-4.16 (m, 2H), 3.85

(s, 6H), 3.72-3.63 (m, 7H), 3.13-3.07 (m, 2H), 2.94-2.83 (m,

1H), 2.65-2.59 (m, 1H), 2.42-2.35 (m, 1H), 2.04-1.98 (m, 1H).

D213
630.15

¹H NMR (300 MHz, Methanol-d4) δ 9.10 (s, 1H), 8.01 (s, 1H), 7.56

(d, J = 8.3 Hz, 1H), 7.27-7.16 (m, 2H), 7.02 (s, 2H), 5.23-5.04

(m, 2H), 4.54-4.43 (m, 4H), 4.43-4.39 (m, 2H), 4.12-3.93 (m,

8H), 3.77 (s, 3H), 3.00-2.86 (m, 1H), 2.85-2.75 (m, 1H), 2.60-

2.42 (m, 1H), 2.25-2.13 (m, 1H).

D214
697.15

¹H NMR (400 MHz, DMSO-d6) δ 10.96 (s, 1H), 7.99 (s, 1H), 7.36

(d, J = 8.0 Hz, 1H), 6.88 (s, 2H), 6.68 (d, J = 8.1 Hz, 2H), 5.08 (dd,

J = 13.3, 5.1 Hz, 1H), 4.38-4.13 (m, 2H), 3.85 (s, 6H), 3.63 (s,

3H), 3.57 (d, J = 4.5 Hz, 6H), 2.97-2.83 (m, 1H), 2.77 (s, 3H),

2.63-2.54 (m, 2H), 2.37 (dd, J = 13.2, 4.6 Hz, 2H), 2.02-1.94

(m, 1H), 1.73 (t, J = 5.5 Hz, 4H).

D215
683.2

¹H NMR (400 MHz, Methanol-d4) δ 9.47 (s, 1H), 7.99 (s, 1H), 7.38

(d, J = 8.2 Hz, 1H), 7.19 (s, 2H), 6.86 (d, J = 2.2 Hz, 1H), 6.77 (m,

J = 8.2, 2.3 Hz, 1H), 5.16-5.07 (m, 1H), 4.37 (d, J = 7.8 Hz, 2H),

4.30 (s, 2H), 3.98 (s, 6H), 3.78 (s, 3H), 3.74 (s, 4H), 3.24 (s, 4H),

2.96-2.83 (m, 1H), 2.83-2.73 (m, 1H), 2.56-2.40 (m, 1H), 2.21-

2.09 (m, 5H).

D216
682.3

¹H NMR (300 MHz, DMSO-d6) δ 10.97 (s, 1H), 8.55 (d, J = 3.2 Hz,

1H), 7.89 (d, J = 3.2 Hz, 1H), 7.44-7.27 (m, 2H), 6.84 (s, 2H),

6.76-6.62 (m, 2H), 5.08 (dd, J = 13.1, 5.1 Hz, 1H), 4.38-4.12

(m, 2H), 3.86 (s, 6H), 3.67-3.53 (m, 6H), 3.50 (s, 3H), 3.02-

2.80 (m, 1H), 2.66-2.52 (m, 4H), 2.43-2.26 (m, 2H), 2.06-1.91

(m, 1H), 1.75 (s, 4H).

D217
667.45

¹H NMR (300 MHz, Methanol-d4) δ 8.74 (s, 1H), 8.16 (s, 1H), 7.42

(d, J = 8.2 Hz, 1H), 7.38 (s, 2H), 6.89 (d, J = 2.1 Hz, 1H), 6.81 (dd,

J = 8.2, 2.3 Hz, 1H), 5.14 (dd, J = 13.3, 5.1 Hz, 1H), 4.45-4.36

(m, 4H), 4.05 (s, 6H), 3.91-3.68 (m, 7H), 3.62-3.40 (m, 2H),

3.29-3.12 (m, 2H), 3.01-2.74 (m, 2H), 2.50 (qd, J = 13.0, 4.7

Hz, 1H), 2.39-1.98 (m, 5H).

D218
747.45

¹H NMR (300 MHz, Methanol-d4) δ 8.59 (s, 1H), 8.24 (s, 1H, FA),

7.86 (s, 1H), 7.56 (d, J = 8.3 Hz, 1H), 7.48-7.37 (m, 2H), 7.30 (s,

2H), 5.23 (ddd, J = 13.2, 5.2, 2.2 Hz, 1H), 4.71 (dd, J = 15.5, 7.7

Hz, 1H), 4.53-4.41 (m, 4H), 4.10 (d, J = 1.1 Hz, 7H), 3.98 (d, J =

13.1 Hz, 1H), 3.84 (s, 3H), 3.32 (d, J = 13.3 Hz, 1H), 3.20 (d, J =

6.3 Hz, 1H), 3.05-2.96 (m, 7H), 2.92-2.82 (m, 1H), 2.58 (qd, J =

13.1, 4.7 Hz, 1H), 2.33-2.09 (m, 3H).

D219
654.4

¹H NMR (300 MHz, DMSO-d6) δ 8.08 (s, 1H), 7.68 (s, 1H), 7.50

(d, J = 8.3 Hz, 1H), 7.36 (d, J = 14.9 Hz, 2H), 6.93 (d, J = 2.2 Hz,

2H), 6.51 (s, 1H), 5.03 (dd, J = 13.4, 5.1 Hz, 1H), 4.43-4.20 (m,

2H), 4.06-3.93 (m, 3H), 3.78 (s, 6H), 3.59 (s, 3H), 3.01 (t, J =

12.1 Hz, 2H), 2.88-2.72 (m, 1H), 2.70-2.62 (m, 1H), 2.43-2.24

(m, 1H), 1.97 (dd, J = 22.7, 10.4 Hz, 3H), 1.73 (t, J = 10.8 Hz, 2H).

D220
794.7

¹H NMR (400 MHz, Methanol-d4) δ 7.78-7.69 (m, 3H), 7.61 (d,

J = 5.4 Hz, 1H), 7.08 (s, 2H), 7.06-6.97 (m, 2H), 5.14 (dd, J = 13.4,

5.2 Hz, 1H), 4.55-4.37 (m, 4H), 4.02 (s, 6H), 3.76 (s, 3H), 3.65

(d, J = 12.5 Hz, 2H), 3.59-3.53 (m, 1H), 3.40-3.33 (m, 1H), 3.28-

3.16 (m, 2H), 3.10 (d, J = 6.7 Hz, 2H), 3.04-2.87 (m, 3H), 2.84-

2.75 (m, 1H), 2.74-2.64 (m, 1H), 2.57-2.41 (m, 2H), 2.36-

2.21 (m, 2H), 2.21-1.93 (m, 9H), 1.71-1.57 (m, 2H).

DD1
775.10

¹H NMR (300 MHz, DMSO-d6) δ 12.39 (s, 1H), 9.05-8.89 (m,

2H), 8.61 (d, J = 8.4 Hz, 2H), 8.32 (dd, J = 6.4, 2.9 Hz, 1H), 7.96

(s, 1H), 7.57 (s, 1H), 7.36-7.24 (m, 2H), 6.90 (s, 2H), 4.87 (d, J =

47.4 Hz, 2H), 4.55 (s, 1H), 3.83 (s, 7H), 3.62 (s, 4H), 3.46 (s, 2H),

2.13 (s, 8H).

DD2
654.20

¹H NMR (300 MHz, Methanol-d4) δ 9.05 (s, 1H), 8.44-8.36 (m,

2H), 7.74-7.67 (m, 2H), 7.61-7.50 (m, 2H), 7.36-7.27 (m, 2H),

7.00 (s, 2H), 4.12-3.86 (m, 12H), 3.75 (s, 3H), 3.11-2.98 (m,

4H).

Example 31—BRD9 Bromodomain TR-FRET Competition Binding Assay

This example demonstrates the ability of the compounds of the disclosure to biochemically inhibit BRD9 bromodomain in a competition binding assay.

Procedure: His-Flag-BRD9 (P133-K239; Swiss Prot Q9H8M2; SEQ ID NO:1 mgsshhhhhhenlyfq/gdykddddkgslevlfqg/PAENESTPIQQLLEHFLRQLQRKDPHGFFAFPVTDAIAPGYSMII KHPMDFGTMKDKIVANEYKSVTEFKADFKLMCDNAMTYNRPDTVYYKLAKKILHAGFKMMSK) was cloned, expressed, purified, and then treated with TEV protease. Cleaved His tag was removed by purification. The binding of a biotinylated small molecule ligand of BRD9 was assessed via the LANCE® TR-FRET platform (PerkinElmer), and the compounds were assayed for inhibitory activity against this interaction.

Results: A mixture of biotinylated-ligand and SureLight™ Allophycocyanin-Streptavidin (APC-SA, PerkinElmer AD0201) in 50 mM HEPES (pH 7.4), 50 mM NaCl, 1 mM TCEP (pH 7), 0.01% (v/v) Tween-20, 0.01% (w/v) bovine serum albumin was added to a white 384-well PerkinElmer Proxiplate Plus plate. DMSO or 3-fold serially diluted compounds were then added to the Proxiplate followed by addition of Flag-BRD9. After a 10-minute incubation at room temperature, Eu-W1024 anti-FLAG (PerkinElmer, AD0273) was added. The final reaction mixture that contained 3.75 nM biotinylated ligand, 3 nM Flag-BRD9, 7.5 nM SureLight™ Allophycocyanin-Streptavidin, and 0.2 nM Eu-W1024 anti-FLAG was incubated at room temperature for 90 minutes.

The plates were then read on a PerkinElmer Envision plate reader to determine the ratio of emission at 665 nm over 615 nm. Data was normalized to a DMSO control (100%) and a no protein control (0%) and then fit to a four parameter, non-linear curve fit to calculate an IC₅₀(μM) as shown in Table 5. As shown by the results in Table 5, a number of compounds of the present disclosure exhibit an IC₅₀value of <1 μM for BRD9 binding, indicating their affinity for targeting BRD9.

TABLE 5

Bromodomain TR-FRET Binding

Compound No.
Bromodomain TR-FRET BRD9 IC₅₀(nM)

B1
+++

B2
++++

B3
+++

B4
+++

B5
++++

B6
++++

B7
++

B8
+++

B9
+++

B10
+++

B11
++

D1
++

D2
++++

D3
+++

D4
+++

D5
++

D6
++++

D7
++

D8
++

D9
+++

D10
++++

D11
++++

D12
++++

D13
++++

D14
+++

D15
++++

“+” indicates inhibitory effect of ≥1000 nM;

“++” indicates inhibitory effect of ≥100 nM;

“+++” indicates inhibitory effect of ≥10 nM;

“++++” indicates inhibitory effect of <10 nM;

“NT” indicates not tested

Example 32—SYO1 BRD9 NanoLuc Degradation Assay

This example demonstrates the ability of the compounds of the disclosure to degrade a Nanoluciferase-BRD9 fusion protein in a cell-based degradation assay.

Procedure: A stable SYO-1 cell line expressing 3×FLAG-NLuc-BRD9 was generated. On day 0 cells were seeded in 30 μL media into each well of 384-well cell culture plates. The seeding density was 8000 cells/well. On day 1, cells were treated with 30 nL DMSO or 30 nL of 3-fold serially DMSO-diluted compounds (10 points in duplicates with 1 μM as final top dose). Subsequently plates were incubated for 6 hours in a standard tissue culture incubator and equilibrated at room temperature for 15 minutes. Nanoluciferase activity was measured by adding 15 μL of freshly prepared Nano-Glo Luciferase Assay Reagent (Promega N1130), shaking the plates for 10 minutes and reading the bioluminescence using an EnVision reader.

Results: The Inhibition % was calculated using the following formula: % Inhibition=100×(Lum_HC−Lum_Sample)/(Lum_HC−Lum_LC). DMSO treated cells are employed as High Control (HC) and 1 μM of a known BRD9 degrader standard treated cells are employed as Low Control (LC). The data was fit to a four parameter, non-linear curve fit to calculate IC₅₀(μM) values as shown in Table 6A and Table 6B. As shown by the results in Table 6A and Table 6B, a number of compounds of the present disclosure exhibit an IC₅₀value of <1 μM for the degradation of BRD9, indicating their use as compounds for reducing the levels and/or activity of BRD9 and their potential for treating BRD9-related disorders.

TABLE 6A

SYO1 BRD9-NanoLuc Degradation

Compound No.
SYO1 BRD9-NanoLuc degradation IC₅₀(nM)

D1
+

D2
+++

D3
++++

D4
+++

D5
+

D6
+

D7
++

D8
++

D9
+

D10
++++

D11
+

D12
++++

D13
++++

D14
++

D15
++++

“+” indicates inhibitory effect of ≥1000 nM;

“++” indicates inhibitory effect of ≥100 nM;

“+++” indicates inhibitory effect of ≥10 nM;

“++++” indicates inhibitory effect of <10 nM;

“NT” indicates not tested

TABLE 6B

SYO1 BRD9-NanoLuc Degradation

Compound No.
SYO1 BRD9-NanoLuc degradation IC₅₀(nM)

D16
++++

D17
++++

D18
++++

D19
++++

D20
++++

D21
++++

D22
++++

D23
++++

D24
++++

D25
++++

D26
++++

D27
++++

D28
++++

D29
++++

D30
+++

D31
+++

D32
++

D33
++++

D34
++

D35
++++

D36
++++

D37
++++

D38
++++

D39
++++

D40
++++

D41
++++

D42
++++

D43
++++

D44
+++

D45
+

D46
++++

D47
+++

D48
+++

D49
++++

D50
++++

D51
+++

D52
+++

D53
+++

D54
++

D55
++

D56
+++

D57
++++

D58
+++

D59
+++

D60
++++

D61
++

D62
+++

D63
++

D64
++

D65
++++

D66
++++

D67
+++

D68
+++

D69
+++

D70
++++

D71
+++

D72
++++

D73
++++

D74
++++

D75
++++

D76
+++

D77
++++

D78
++++

D79
++++

D80
++

D81
++

D82
++

D83
++++

D84
++++

D85
++++

D86
++++

D87
++++

D88
+++

D89
++++

D90
++++

“+” indicates inhibitory effect of ≥1000 nM;

“++” indicates inhibitory effect of ≥100 nM;

“+++” indicates inhibitory effect of ≥10 nM;

“++++” indicates inhibitory effect of <10 nM;

“NT” indicates not tested

TABLE 6C

SYO1 BRD9-NanoLuc Degradation

Compound No.
SYO1 BRD9-NanoLuc degradation IC₅₀(nM)

D91
++++

D92
++++

D93
++++

D94
++++

D95
++

D96
++++

D97
++++

D98
++++

D99
++++

D100
++++

D101
++++

D102
++++

D103
++++

D104
++++

D105
++++

D106
++++

D107
++++

D108
++++

D109
++++

D110
++++

D111
++++

D112
++++

D113
++++

D114
++++

D115
++++

D116
++++

D117
++++

D118
++++

D119
++++

D120
++++

D121
++++

D122
++++

D123
++++

D124
++++

D125
++++

D126
++++

D127
++++

D128
++++

D129
++++

D130
++++

D131
++++

D132
++++

D133
++++

D134
++++

D135
++++

D136
++++

D137
++

D138
++++

D139
++++

D140
++++

D141
++++

D142
+

D143
+

D144
++++

D145
++

D146
++++

D147
++++

D148
+

D149
++++

D150
++++

D151
++++

D152
++++

D153
++++

D154
++++

D155
++++

D156
++++

D157
++++

D158
++++

D159
++++

D160
++++

D161
++++

D162
++++

D163
++++

D164
++++

D165
++++

D166
++++

D167
++++

D168
++++

D169
++++

D170
++++

D171
+++

D172
+

D173
++++

D174
++++

D175
++++

D176
++++

D177
++++

D178
++++

D179
++++

D180
++++

D181
++++

D182
++++

D183
++++

D184
++++

D185
++++

D186
++++

D187
++++

D188
++++

D189
++++

D190
++++

D191
++++

D192
++++

D193
++++

D194
++++

D195
++++

D196
++++

D197
++++

D198
+++

D199
++++

D200
++++

D201
++++

D202
++++

D203
++++

D204
++

D205
++++

D206
++++

D207
++++

D208
++++

D209
++++

D210
++++

D211
+++

D212
++++

D213
++++

D214
++++

D215
++++

D216
++++

D217
++++

D218
+++

D219
++++

D220
++++

DD1
+

DD2
+

“+” indicates inhibitory effect of ≥1000 nM;

“++” indicates inhibitory effect of ≥100 nM;

“+++” indicates inhibitory effect of ≥10 nM;

“++++” indicates inhibitory effect of <10 nM;

“NT” indicates not tested

Other Embodiments

All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference in their entirety to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference in its entirety. Where a term in the present application is found to be defined differently in a document incorporated herein by reference, the definition provided herein is to serve as the definition for the term.

While the invention has been described in connection with specific embodiments thereof, it will be understood that invention is capable of further modifications and this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure that come within known or customary practice within the art to which the invention pertains and may be applied to the essential features hereinbefore set forth, and follows in the scope of the claims.

Other embodiments are in the claims.

	Number	Date	Country
	62881128	Jul 2019	US
	62798418	Jan 2019	US

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