OTUB1 small-molecule binders and OTUB1-recruiting deubiquitinase-targeting chimeras (dubtacs)

Abstract

Disclosed are compounds that are irreversible small-molecule binders of OTUB1 (also known as Otubain 1), and bivalent compounds (e.g. bi-functional small molecule compounds) which recruit OTUB1 through the irreversible small-molecule binders of OTUB1 to de-ubiquitinylate and stabilize proteins such as AMPK, cGAS and CFTR. Also disclosed are methods for the treatment of AMPK, cGAS and CFTR-mediated diseases in a subject in need thereof. Also disclosed are methods for designing such compounds including irreversible small-molecule binders of OTUB1 and related bivalent compounds as de-ubiquitinase-targeting chimeras (DUBTACs) for targeted protein stabilization.

Description

[0001] OTUB1 Small-Molecule Binders and OTUBl-recruiting Deubiquitinase- targeting Chimeras (DUBTACs)

[0002] STATEMENT REGARDING GOVERNMENT FUNDING

[0003] This invention was made with government support under R35CA253027 awarded by the National Institutes of Health. The government has certain rights in the invention.

[0004] CROSS-REFERENCE TO RELATED APPLICATION

[0005] The present application claims priority to U.S. Provisional Application Serial No. 63 / 497,651, filed on April 21, 2023, the contents of which are hereby incorporated by reference in their entirety.

[0006] TECHNICAL FIELD

[0007] The present disclosure is directed to compounds that are irreversible small-molecule binders of OTUB1 (also known as Otubain 1), and bivalent compounds (e.g. bi-functional small molecule compounds) which recruit OTUB1 through the irreversible small-molecule binders of OTUB1 to de-ubiquitinylate and stabilize proteins such as AMPK, cGAS and CFTR. The present disclosure is also related to methods for the treatment of AMPK, cGAS and CFTR-mediated diseases in a subject in need thereof. The disclosure also relates to methods for designing such compounds including irreversible small-molecule binders of OTUB1 and related bivalent compounds as deubiquitinase-targeting chimeras (DUBTACs) for targeted protein stabilization (TPS).

[0008] BACKGROUND

[0009] Protein ubiquitination plays a critical role in a variety of cellular process, like protein degradation, quality control, trafficking, and signaling. Therefore, controlling the ubiquitination level of disease related proteins can achieve substantial therapeutic effects. For example, inducing the ubiquitination of oncoproteins such as Myc and Ras to lead to their degradation would greatly benefit related cancer therapy. Similarly, deubiquitination and stabilization of proteins such as tumor suppressors would also be greatly beneficial for the treatment of relevant diseases including cancer.

[0010] 5' adenosine monophosphate-activated protein kinase (also known as AMPK) is an enzyme that plays key roles in cellular energy homeostasis. Recent studies identified AMPK as regulating diverse metabolic and physiological processes. However, it is downregulated in major chronic diseases, such as obesity, inflammation, diabetes and cancer. Therefore, stabilization of AMPK in the cellular context may enhance AMPK activation, and has the potential to provide a novel therapeutic strategy to treat such diseases (Jeon, 2016).

[0011] The protein stabilization strategy could also be applied to other targets, such as cyclic GMP-AMP (cGAMP) synthase (also known as cGAS). The cGAS-stimulator of interferon genes (STING) pathway is a component of the innate immune system. cGAS detects the presence of cytosolic DNA and, in response, triggers expression of inflammatory genes that can lead to senescence or to the activation of defense mechanisms. In tumor cells, the cGAS-STING pathway exerts its antitumor effects through the induction of a robust type I IFN response, which activates immune cells, particularly dendritic cells (DCs), in the tumor micro environment (TME) (Samson and Ablasser, 2022).

[0012] In addition to AMPK and cGAS, Cystic fibrosis transmembrane conductance regulator (CFTR) is another suitable protein target for stabilization. CFTR AF508 mutation destabilizes CFTR, and leads to the cystic fibrosis phenotype (Ward et al., 1995). Therefore, deubiquitination of AF508 mutant CFTR could be an effective strategy to prevent the cystic fibrosis process.

[0013] The targeted protein degradation (TPD) field has made great progress over the past 10 years. PROteolysis TArgeting Chimera (PROTAC) technology has been widely utilized in the drug discovery field (Dale et al., 2022). Deubiquitinase-targeting chimera (DUBTAC), which hijacks a cellular deubiquitinase to remove the polyubiquitin attached to the target protein by inducing close proximity between the target protein and deubiquitinase, recently emerged as an effective technology to improve target protein stability by reducing the proteasomal degradation of the target protein (Henning et al., 2022; Liu et al., 2022). DUBTACs provide a novel therapeutic approach for treating various diseases.

[0014] SUMMARY The present disclosure includes two parts: (A) small molecules that covalently bind the deubiquitinase OTUB1, and (B) heterobifunctional small molecules that stabilize AMPK, cGAS, or CFTR by recruiting OTUB 1.

[0015] A. OTUB1 irreversible small-molecule binders

[0016] OTUB1 (also known as OUT domain-containing ubiquitin aldehyde-binding protein 1) is a K48 linkage-specific deubiquitinating enzyme. There are very few small molecules reported to bind OTUB1, including the covalent binder EN523 (Henning et al., 2022), and reversible inhibitor compound 61 (Tan et al., 2022), ML364 (WO2020223551A1). However, EN523 has low binding affinity to OTUB1, and compound 61 and ML364 inhibit the enzymatic activity of OTUB1. Therefore, development of more effective OTUB1 small-molecule binders that lead to effective DUBTACs is needed.

[0017] Described herein are small molecules that covalently modify OTUB1.

[0018] In one aspect, the present disclosure features small molecules that covalently modify OTUB1 of Formula (A-I) below:

[0019] Formula (A-I) or a tautomer thereof, or a pharmaceutically acceptable salt of the compound or the tautomer, wherein

[0020] ADis selected from N, or CRD2;

[0021] BD is selected from C=O, CH2, CH RD2, or C(RD2)2;

[0022] Ring CDis absent, or selected from C3-C12cycloalkyl, 3-12-membered heterocyclic, C6- C10aryl, and 5-10 membered heteroaryl;

[0023] Ring DDis a saturated or partially unsaturated 4-12 membered heterocyclic;

[0024] FD is selected from N, or CRD2;

[0025] LD1is a bond, or a bivalent group selected from -O-, -NRD5-, -C(O)-, -C(O)O-, -C(O)NRD5-, -NRD5C(O)-, -OC(O)NRD5-, -NRD5C(O)O-, -NRD6C(O)NRD5-, -S(O)-, -S(O)2-, -S(O)NRD5-, - S(O)2NRD5-, CI-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, and C2-C6alkynylene;

[0026] LD2is selected from -C(O)-, -S(O)-, -S(O)2-, -NRD5C(O)-, and C1-C6alkylene;

[0027] LD3is a bond, or a bivalent group selected from C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, and C2-C6alkynylene;

[0028] RD1is selected from C1-C6haloalkyl, C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2-C6heteroalkynyl, partially unsaturated C4-C8 cycloalkyl, and partially unsaturated 4-8 membered heterocyclic, where each said C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2- C6, heteroalkynyl, partially unsaturated C4-C cycloalkyl, and partially unsaturated 4-8 membered heterocyclic were optionally substituted with hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O)NRD7RD8, C(O)ORD7, tri(Ci-C3alkyl) silyl, C1-C6alkyl, C1-C6heteroalkyl, and 3-8 membered heterocyclic; each RD2is independently selected from hydrogen, halogen, cyano, ORD7, NRD7RD8, Ci- C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6heteroalkenyl, C1-C6alkynyl, C2-C6heteroalkynyl, C3-C8 cycloalkyl, and 3-8 membered heterocyclic; or two RD2groups, together with the atom(s) to which they are connected, optionally form a C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, where each said C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, together with Ring DD, can optionally form fused rings or bridged rings; each RD3is selected from hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O)ORD7, C(O)NRD7RD8, -OC(O)NRD7RD8, - NRD9C(O)NRD7RD8-, -S(O)NRD7RD8-, -S(O)2NRD7RD8-, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C1-C6alkynyl, C3-C12cycloalkyl, 3-12- membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl; or two RD3groups, together with the atom(s) to which they are connected, optionally form a C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, where each said C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, together with Ring CD, optionally form fused rings or bridged rings;

[0029] RD4is RD4aor RD4b;

[0030] RD4ais selected from hydrogen, ORD7, NRD7RD8, C(O)RD7, C(O)ORD7, C(O)NRD7RD8, - OC(O)NRD7RD8, -NRD9C(O)NRD7RD8-, S(O)RD7, S(O)2RD7, S(O)NRD7RD8, S(O)2NRD7RD8, C1-C6 alkyl, Ci-Ce haloalkyl, Ci-Ce heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C12 cycloalkyl, 3-12- membered heterocyclic, Ce-Cw aryl, and 5-10 membered heteroaryl;

[0031] Ro4bis a bivalent group that connect to the Linker moiety in Formula (B), and Ro4bis selected from a bond, -O-, -N-, -C(O)-, -C(O)O-, -C(O)NRD7-, -OC(O)NRD7-, -NRD9C(O)NRD7-, -S(O)-, -S(O)2-, -S(O)NRD7-, -S(O)2NRD7-, CI-C6alkylene, Ci-C6haloalkylene, Ci-C6heteroalkylene, C2-C6 alkenylene, C2-C6 alkynylene, C3-C12 cycloalkylene, 3-12-membered heterocyclicene, Ce-Cio arylene, and 5-10 membered heteroarylene; each RD5and each RD6are independently selected from hydrogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci-Ce heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C12 cycloalkyl, 3-12 membered heterocyclic, Ce-Cio aryl, and 5-10 membered heteroaryl; each RD7, each RD8and each RD9are independently selected from hydrogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci-Ce heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C12 cycloalkyl, 3-12 membered heterocyclic, Ce-Cio aryl, and 5-10 membered heteroaryl; or

[0032] RD7and RD8, together with the atom(s) to which they are connected, optionally form 3-12- membered heterocyclic ring; mDis an integer of 0-8; and nDis an integer of 0-8.

[0033] In some embodiments, LD1is a bond, or a bivalent group selected from -C(O)-, -NRD5C(O)-, and -C(O)1STRD:>- In some embodiments, LD1is a bond. In some embodiments, LD1is -C(O)- In some embodiments, LD1is -C(O)NRD5-. In some embodiments, LD1is -NRD5C(O)-. In some embodiments, LD3is a bond, or a bivalent group selected from -CH2-, or -CH2CH2-. In some embodiments, LD3is a bond. In some embodiments, LD3is a bond -CH2-.

[0034] In some embodiments, the small molecules that covalently modify OTUB 1 are selected from Formulae (A-I-a), (A-I-b), (A-I-c), or (A-I-d):

[0035] Formula (A-I-a), Formula (A-I-b),

[0036] 5

[0037] Formula (A-I-c), or Formula (A-I-d).

[0038] In some embodiments, AD is N. In some embodiments, AD is CRD2. In some embodiments, AD in Formula (A-I-a) is N. In some embodiments, AD in Formula (A-I-b) is CRD2. In some embodiments, AD in Formula (A-I-c) is N. In some embodiments, AD in Formula (A-I-d) is CRD2.

[0039] In some embodiments, the small molecules that covalently modify OTUB1 are selected from Formulae (A-I-al), (A-I-bl), (A-I-cl), or (A-I-dl):

[0040] Formula (A-I-c 1), or Formula (A-I-d 1)

[0041] In some embodiments, the small molecules that covalently modify OTUB1 are selected from Formulae (A-I-al), (A-I-bl), (A-I-cl), and (A-I-dl). In some embodiments, the small molecules that covalently modify OTUB1 comprise Formula (A-I-al). In some embodiments, the small molecules that covalently modify OTUB1 comprise Formula (A-I-bl). In some embodiments, the small molecules that covalently modify OTUB1 comprise Formula (A-I-cl). In some embodiments, the small molecules that covalently modify OTUB1 comprise Formula (A-I- dl).

[0042] 6 In some embodiments, Ring DD in Formulae (A-I-a), (A-I-al), (A-I-bl), (A-I-cl), and (A- I-dl) is selected from 6-8 membered monocyclic heterocyclic, 7-12 membered spiro heterocyclic, 7-12 membered bridged heterocyclic, and 6-12 membered fused heterocyclic.

[0043] In some embodiments, The small molecules that covalently modify OTUB1 are selected from Formulae (A-I-a2), (A-I-a3), (A-I-a4), (A-I-a5), (A-I-b2), (A-I-b3), (A-I-b4), (A-I-b5), (A-I- c2), (A-I-c3), (A-I-c4), (A-I-c5), (A-I-d2), (A-I-d3), (A-I-b4), and (A-I-d5):

[0044] Formula (A-I-d4), Formula (A-I-d5), wherein each ED1, each ED2, each ED3, each ED4, and each ED5are independently selected from null, O, CO, SO, S(O)2, NRD2, and CRD2RD2, with the proviso that no two oxygen atoms are connected to each other;

[0045] OD1is selected from an integer of 1-4; and

[0046] OD2, OD3, OD4, and OD5are independent selected from an integer of 0-4.

[0047] In some embodiments, the small molecules that covalently modify OTUB1 comprise Formulae (A-I-a3), (A-I-b2), and (A-I-c4). In some embodiments, the small molecules that covalently modify OTUB 1 comprise Formula (A-I-a3). In some embodiments, the small molecules that covalently modify 0TUB1 comprise Formula (A-I-b2). In some embodiments, the small molecules that covalently modify OTUB1 comprise Formula (A-I-c4).

[0048] In some embodiments, each ED1, each ED2, each ED3, each ED4, and each ED5are independently selected from null, CO, and CRD2RD2. In some embodiments, each ED1, each ED2, each ED3, each ED4, and each ED5are independently selected from CO and CRD2RD2.

[0049] In some embodiments, ED1, ED2, ED3, ED4, and ED3in Formulae (A-I-a2), (A-I-a3), (A-I- a4), (A-I-a5), (A-I-b2), (A-I-b3), (A-I-b4), (A-I-b5), (A-I-c2), (A-I-c3), (A-I-c4), (A-I-c5), (A-I- d2), (A-I-d3), (A-I-b4), and (A-I-d5) are CRD2RD2.

[0050] In some embodiments, the small molecules that covalently modify OTUB1 are selected from Formulae (A-I-a6), (A-I-a7), (A-I-a8), (A-I-a9), (A-I-b6), (A-I-b7), (A-I-b8), (A-I-b9), (A-I- c6), (A-I-c7), (A-I-c8), (A-I-c9), (A-I-d6), (A-I-d7), (A-I-d8), and (A-I-d9):

[0051] Formula (A-I-d8), and Formula (A-I-d9), wherein

[0052] RD2is a substituent that can attach to anywhere on the monocyclic and bicyclic rings.

[0053] In some embodiments, FD is selected from N, CH, or CMe. In some embodiments, FD is selected from N. In some embodiments, FD is selected from CH. In some embodiments, FD is selected from CMe.

[0054] In some embodiments, LD3is selected from a bond or -CH2-. In some embodiments, LD3is selected from a bond. In some embodiments, LD3is selected from -CH2-. In some embodiments, the small molecules that covalently modify OTUB1 comprise Formulae (A-I-a6), (A-I-a7), (A-I-b6), (A-I-c8), and (A-I-d6). In some embodiments, the small molecules that covalently modify OTUB1 comprise Formula (A-I-a6). In some embodiments, the small molecules that covalently modify OTUB1 comprise Formula (A-I-a7). In some embodiments, the small molecules that covalently modify OTUB1 comprise Formula (A-I-b6). In some embodiments, the small molecules that covalently modify OTUB1 comprise Formula (A-I- c8). In some embodiments, the small molecules that covalently modify OTUB 1 comprise Formula (A-I-d6).

[0055] In some embodiments, BD is selected from C=O and C(RD2)2. In some embodiments, BD is C=O. In some embodiments, BD is C(RD2)2.

[0056] In some embodiments, Ring DDin Formulae (A-I), (A-I-a), (A-I-al), (A-I-b), (A-I-bl), (A- I-c), (A-I-cl), (A-I-d), and (A-I-dl) is selected from Ring DDal, Ring DDa2, Ring DDb1, Ring DDb2, Ring DDC1, and Ring DDC2.

[0057] In some embodiments, Ring DDin Formulae (A-I-a), (A-I-al), (A-I-c), and (A-I-cl) is Ring DDalcomprising RD2substituted groups of:

[0058] In some embodiments, Ring DDin Formulae (A-I-a), (A-I-al), (A-I-c), and (A-I-cl) is Ring DDa2comprising RD2substituted groups of:

[0059]

[0060] In some embodiments, Ring DDin Formulae (A-I-a) and (A-I-al) is selected from

[0061] In some embodiments, the small molecules that covalently modify OTUB1 comprise

[0062] Formula (A-I-alO):

[0063] Formula (A-I-alO).

[0064] In some embodiments, the small molecules that covalently modify OTUB1 comprise Formula (A-I-al 1):

[0065]

[0066] In some embodiments, Ring DDin Formulae (A-I-c) and (A-I-cl) is

[0067] The small molecules that covalently modify OTUB1 comprise Formula (A-I-clO):

[0068] Formula (A-I-c 10).

[0069] In some embodiments, Ring DDin Formulae (A-I-b), (A-I-bl), (A-I-d), and (A-I-dl) is Ring DDb1comprising RD2substituted groups of:

[0070] In some embodiments, Ring DDin Formulae (A-I-b), (A-I-bl), (A-I-d), and (A-I-dl), is Ring DDb2comprising RD2substituted groups of:

[0071]

[0072] In some embodiments, Ring DDin Formulae (A-I-b), (A-I-bl), (A-I-d), and (A-I-dl) is

[0073] In some embodiments, the small molecules that covalently modify OTUB1 comprise

[0074] Formula (A-I-b 10):

[0075] In some embodiments, the small molecules that covalently modify OTUB1 comprise

[0076] Formula (A-I-dlO): In some embodiments, Ring DDin Formulae (A-I-b), (A-I-bl), (A-I-d), and (A-I-dl) is Ring DDb1comprising RD2substituted groups of:

[0077] In some embodiments, Ring DDin Formulae (A-I-b), (A-I-bl), (A-I-d), and (A-I-dl), is

[0078] Ring DDb2comprising RD2substituted groups of:

[0079] In some embodiments, Ring DDin Formulae (A-I-b), (A-I-bl), (A-I-d), and (A-I-dl) is

[0080] In some embodiments, the small molecules that covalently modify OTUB1 comprise Formula (A-I-b 11):

[0081]

[0082] In some embodiments, the small molecules that covalently modify OTUB1 comprise

[0083] Formula (A-I-dl 1):

[0084] In some embodiments, ED1in Formulae (A-I-c2), (A-I-c3), (A-I-c4), and (A-I-c5) that is adjacent to N atom is -C(O)-; and the rest of the ED1, ED2, ED3, ED4, and ED5are CRD2RD2. The small molecules that covalently modify OTUB1 comprise Formulae (A-I-cl l), (A-I-cl2), (A-I- cl 3), and (A-I-C14): wherein

[0085] OD1-1 is selected from an integer of 0-3. In some embodiments, the small molecules that covalently modify OTUB1 comprise Formulae (A-I-cl l) and (A-I-cl2). In some embodiments, the small molecules that covalently modify OTUB1 comprise Formulae (A-I-cl 1). In some embodiments, the small molecules that covalently modify OTUB1 comprise Formulae (A-I-cl2).

[0086] In some embodiments, Ring DDin Formulae (A-I-c) and (A-I-cl) is Ring DDC1comprising RD2substituted groups of:

[0087] In some embodiments, Ring DDin Formulae (A-I-c) and (A-I-cl) is Ring DDC2comprising RD2substituted groups of:

[0088] In some embodiments, Ring DDin Formulae (A-I-c) and (A-I-cl) is

[0089] In some embodiments, the small molecules that covalently modify OTUB1 comprise

[0090] Formulae (A-I-C15) and (A-I-C16):

[0091] Formula (A-I-c 15), and Formula (A-I-c 16).

[0092] In some embodiments, Ring CD is selected from C3-C12cycloalkyl, 3-12-membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl. In some embodiments, Ring CDis C3- C12cycloalkyl. In some embodiments, Ring CD is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. In some embodiments, Ring CD is cyclopentyl. In some embodiments, Ring CD is 3-12-membered heterocyclic. In some embodiments, Ring CD is selected from azetidinyl, pyrrolidinyl, piperidinyl, and piperazinyl. In some embodiments, Ring CD is C6-

[0093] C10aryl. In some embodiments, Ring CD is phenyl. In some embodiments, Ring CD is 5-10 membered heteroaryl. In some embodiments, Ring CD is selected from thiophenyl, benzothiophenyl, tetrahydrobenzothiophenyl, thiazolyl, imidazolyl, furanyl, pyrrolyl, pyrazolyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl, benzofuranyl, indolyl, and indazolyl. In some embodiments, Ring CD is selected from thiophenyl, benzothiophenyl, tetrahydrobenzothiophenyl, benzofuranyl, and indolyl. In some embodiments, Ring CD is thiophenyl. In some embodiments, In some embodiments, Ring CD is In some embodiments, Ring CD is benzothiophenyl. In some embodiments, Ring some embodiments, Ring CDis tetrahydrobenzothiophenyl. In some embodiments, Ring

[0094] In some embodiments, LD2is selected from -C(O)-, C1-C6alkylene, and -NRD5C(O)-. In some embodiments, LD2is selected from -C(O)-, -NHC(O)-, and methylene. In some embodiments, LD2is -C(O)-. In some embodiments, -NHC(O)-. LD2is In some embodiments, LD2is methylene.

[0095] In some embodiments, LD2is -C(O)-; and RD1is selected from C1-C6haloalkyl, C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2-C6heteroal kynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic, where each said C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2-C6heteroal kynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic were optionally substituted with hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O)NRD7RD8, C(O)ORD7, tri(Ci-C3alkyl) silyl, C1-C6alkyl, Ci- Ce heteroalkyl, and 3-8 membered heterocyclic. In some embodiments, LD2is C1-C6alkylene; and RD1is selected from C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2-C6heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic, where each said C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2-C6heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic were optionally substituted with hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O)NRD7RD8, C(O)ORD7, tri(C1- C3alkyl) silyl, C1-C6alkyl, C1-C6heteroalkyl, and 3-8 membered heterocyclic. In some embodiments, LD2is methylene; and RD1is selected from C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2-C6heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic, where each said C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2-C6heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic were optionally substituted with hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O)NRD7RD8, C(O)ORD7, tri(C1-C3alkyl) silyl, C1-C6hCet1e-Cro6alkyl, and 3-8 membered heterocyclic.

[0096] In some embodiments, LD2is -NHC(O)-; and RD1is selected from C1-C6haloalkyl, C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2-C6heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic, where each said C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2-C6heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic were optionally substituted with hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O)NRD7RD8, C(O)ORD7, tri(Ci-C3alkyl) silyl, C1-C6alkyl, C1- C6heteroalkyl, and 3-8 membered heterocyclic. In some embodiments, LD2is C1-C6alkylene; and RD1is selected from C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2-C6heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 memberedheterocyclic, where each said C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2-C6heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic were optionally substituted with hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O) NRD7RD8, C(O)ORD7, tri(Ci- C3alkyl) silyl, C1-C6alkyl, C1-C6heteroalkyl, and 3-8 membered heterocyclic. In some embodiments, LD2is methylene; and RD1is selected from C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2-C6heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic, where each said C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2-C6heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic were optionally substituted with hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O)NRD7RD8, C(O)ORD7, tri(Ci-C3alkyl) silyl, aClk1-yCl,6hCet1e-Cro6alkyl, and 3-8 membered heterocyclic.

[0097] In some embodiment Rs,D1is C1-C6haloalkyl. In some embodiments, RD1is halomethyl. In some embodiment Rs,D1is CH2C1 or CH2F. In some embodiments, RD1is selected from C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2-C6heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic, where each said C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2-C6heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic were optionally substituted with hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O)NRD7RD8, C(O)ORD7, tri(Ci-C3alkyl) silyl, C1-C6alkyl, C1- C6heteroalkyl, and 3-8 membered heterocyclic. In some embodiment Rs,D1is selected from C2-C3alkenyl, C2-C3alkynyl, partially unsaturated C4-C6cycloalkyl, and partially unsaturated 4-6 membered heterocyclic, where each said C2-C6alkenyl, C2-C6alkynyl, partially unsaturated C4-C6cycloalkyl, and partially unsaturated 4-6 membered heterocyclic were optionally substituted with hydrogen, halogen, cyano, NRD7RD8, C(O)NRD7RD8, C(O)ORD7, tri(C1-C3alkyl) silyl, C1-C6alkyl, 11-C6heteroalkyl, and 3-8 membered heterocyclic. In some embodiment Rs,D1is selected from vinyl, propylenyl, ethynyl, propargyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, dihydrofuranyl, dihydropyrrolyl, dihydropyranyl, and tertrahydopyridinyl, where each said vinyl, propylenyl, ethynyl, propargyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, dihydrofuranyl, dihydropyrrolyl, dihydropyranyl, and tertrahydopyridinyl were optionally substituted with hydrogen, halogen, cyano, NRD7RD8, C(O)NRD7RD8, C(O)ORD7, trimethyl silyl, C1-C3alkyl, C1-C3heteroalkyl, and 3-6 membered heterocyclic. In some embodiment Rs,D1is selected from vinyl and propylenyl, where each said vinyl and propylenyl were optionally substituted with hydrogen, halogen, cyano, NRD7RD8, C(O)NRD7RD8, C(O)ORD7, trimethyl silyl, C1-C3alkyl, C1-C3heteroalkyl, and 3-6 membered heterocyclic. In some embodiments, RD1is selected from

[0098] In some embodiments, - LD2-RD1is selected from

[0099]

[0100] In some embodiments, - LD2-RD1is selected from

[0101] , D . In some embodiments, - LD2- In some embodiments, each RD2is independently selected from hydrogen, halogen, cyano, ORD7, NRD7RD8, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6heteroalkenyl, C1-C6alkynyl, C2-C6heteroalkynyl, C3-C8cycloalkyl, and 3-8 membered heterocyclic. In some embodiments, each RD2is selected from hydrogen, halogen, hydroxy, C1-C6alkoxy, amino, C1-C6alkyl amino, (C1-C6alkyl)( C1-C6alkyl) amino, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C8 cycloalkyl, 3-8 membered heterocyclic. In some embodiments, each RD2is selected from hydrogen, halogen, C1-C6alkyl, C1-C6haloalkyl, C3-C8cycloalkyl, 3-8 membered heterocyclic. In some embodiments, RD2is hydrogen. In some embodiments, RD2is halogen. In some embodiments, RD2is C1-C6alkyl.

[0102] In some embodiments, two RD2groups, together with the atom(s) to which they are connected, optionally form a C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, where each said C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, together with Ring DD, can optionally form fused rings or bridged rings. In some embodiments, two RD2groups, together with the atom(s) to which they are connected and Ring DD, optionally form a bridged 3-12-membered heterocyclic ring. In some embodiments, two RD2groups, together with the atom(s) to which they are connected and Ring DD, optionally form a bridged 7-membered heterocyclic ring. In some embodiments, two RD2groups, together with the atom(s) to which they are connected and Ring DD, optionally form a bridged 8-membered heterocyclic ring.

[0103] In some embodiments, each RD3is independently selected from hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O)ORD7, C(O)NRD7RD8, -OC(O)NRD7RD8, - NRD9C(O)NRD7RD8-, - S(O)NRD7RD8-, -S(O)2NRD7RD8-, CI-C6alkyl, C h1a-Clo6alkyl, C he1-tCer6oalkyl, C2-C6alkenyl, C1-C6alkynyl, C3-C12cycloalkyl, 3-12-membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl. In some embodiments, each RD3is selected from hydrogen, halogen, cyano, ORD7, NRD7RD8, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, 3-12-membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl. In some embodiments, each RD3is selected from methyl, ethyl, isopropyl, piperidinyl, piperazinyl, and phenyl.

[0104] In some embodiments, two RD3groups, together with the atom(s) to which they are connected, optionally form a C3-C12cycloalkyl ring, or 3-12-membered heterocyclic ring, where each said C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, together with Ring CD, optionally form fused rings or bridged rings. In some embodiments, RD4is RD4aIn some embodiments, RD4ais selected from hydrogen, ORD7, NRD7RD8, C(O)NRD7RD8, S(O)2NRD7RD8, CI-C6alkylene, C1- hCa6loalkylene, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, 3-12-membered heterocyclic, Ce-Cio aryl, and 5-10 membered heteroaryl. In some embodiments, RD4is Ro4b. In some embodiments, RD4is a bivalent group that connect to the Linker moiety in Formula (B), and Ro4bis selected from a bond, -O-, -N-, -C(O)-, -C(O)O-, -C(O)NRD7-, -S(O)2-, -S(O)2NRD7-, C1-C al6kylene, Ci-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, C2-C6alkynylene, C3-C12cycloalkylene, 3- 12-membered heterocyclicene, C6-C10arylene, and 5-10 membered heteroarylene. In some embodiments, RD41is a bond.

[0105] In some embodiments, each RD5and each RD6are independently selected from hydrogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic.

[0106] In some embodiments, each RD7, each RD8and each RD9are independently selected from hydrogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic.

[0107] In some embodiments, RD7and RD8, together with the atom(s) to which they are connected, optionally form 4-6-membered heterocyclic ring.

[0108] In some embodiments, mDis an integer of 0-4. In some embodiments, mDis an integer of 1-3. In some embodiments, OD1is selected from an integer of 1-3. In some embodiments, OD2is selected from an integer of 0-3. In some embodiments, OD3is selected from an integer of 0-3. In some embodiments, OD4is selected from an integer of 0-3. In some embodiments, OD5is selected from an integer of 0-3.

[0109] In some embodiments, the small molecules that covalently modify OTUB1 are selected from the structures of Formulae (A-I), (A-I-a), (A-I-al), (A-I-a2), (A-I-a3), (A-I-a4), (A-I-a5), (A- I-a6), (A-I-a7), (A-I-a8), (A-I-a9), (A-I-alO), (A-I-al 1), (A-I-b), (A-I-bl), (A-I-b2), (A-I-b3), (A- I-b4), (A-I-b5), (A-I-b6), (A-I-b7), (A-I-b8), (A-I-b9), (A-I-blO), (A-I-bl l), (A-I-c), (A-I-cl), (A- Lc2), (A-I-c3), (A-I-c4), (A-I-c5), (A-I-c6), (A-I-c7), (A-I-c8), (A-I-c9), (A-I-clO), (A-I-cl l), (A- I-cl2), (A-I-C13), (A-I-C14), (A-I-C15), (A-I-C16), (A-I-d), (A-I-dl), (A-I-d2), (A-I-d3), (A-I-d4), (A-I-d5), (A-I-d6), (A-I-d7), (A-I-d8), (A-I-d9), (A-I-dlO), and (A-I-dl 1), wherein RD4is RD4aIn some embodiments, the small molecules that covalently modify OTUB1 are selected from the structures of Formulae (A-I), (A-I-a), (A-I-al), (A-I-a2), (A-I-a3), (A-I-a4), (A-I-a5), (A-I-a6), (A- I-a7), (A-I-a8), (A-I-a9), (A-I-alO), (A-I-al l), (A-I-b), (A-I-b 1), (A-I-b2), (A-I-b3), (A-I-b4), (A- I-b5), (A-I-b6), (A-I-b7), (A-I-b8), (A-I-b9), (A-I-b 10), (A-I-b 11), (A-I-c), (A-I-cl), (A-I-c2), (A- I-c3), (A-I-C4), (A-I-C5), (A-I-C6), (A-I-c7), (A-I-c8), (A-I-c9), (A-I-c 10), (A-I-c 11), (A-I-c 12), (A-I-C13), (A-I-C14), (A-I-C15), (A-I-d), (A-I-dl), (A-I-d2), (A-I-d3), (A-I-d4), (A-I-b5), (A-I-b6), (A-I-b 10), (A-I-d 11), wherein RD4is Ro4a; and exclude

[0110] In some embodiments, the small molecules that covalently modify OTUB1 are selected from XS154-91, XS154-130, XS154-148, XS154-114, XS154-149, XS159-13, XS159-107, XS165-30, XS159-90, XS165-53, XS165-38, XS165-33, XS165-54, XS154-184, XS165-75, XS165-77, XS165-127, XS165-106, XS165-112, XS165-97, XS165-118, XS165-110, XS165-119, XS165-123, XS165-126, XS165-120, XS165-121, XS165-100, XS165-99, XS165-113, XS165- 109, XS165-117, XS165-105, XS165-154, XS165-170, XS165-155, XS165-172, XS165-169, XS165-177, XS175-45, XS175-46, XS175-59, XS175-63, XS175-64, XS175-67, XS175-68, XS175-70, XS175-71, XS175-76, XS175-110, XS175-120, XS175-126, XS175-132, XS175-133, XS175-137, XS175-143, XS175-148, XS175-149, XS175-158, XS175-159, XS175-160, XS175- 173, XS175-174, XS175-175, XS175-176, XS175-178, XS175-179, XS175-180, XS175-186, XS186-3, XS185-4, XS186-5, XS185-6, XS185-24, XS185-29, XS185-43, XS185-46, XS185-59, XS185-64, XS185-65, XS185-66, XS185-67, XS185-69, XS185-70, XS185-71, XS185-72, XS185-75, XS185-78, XS185-86, XS185-87, XS185-90, 91, XS185-96, XS185-97, XS185-101, XS185-109, XS185-113, XS185-114, XS185-116, XS185-118, XS185-122, XS185-131, XS185- 134, XS185-135, XS185-138, XS185-140, XS185-147, XS185-149, XS185-150, XS185-171, XS190-9, XS190-27, XS190-38, XS190-44, XS190-45, XS190-46, XS190-47, XS190-48, XS190-49, XS190-50, XS190-59, XS190-68, XS190-69, XS190-70, XS190-75, XS190-75-2, XS190-77, XS190-80, XS190-81, XS190-126, XS190-127, XS190-128, XS190-129, XS190-130, XS190-137, XS190-138, XS190-144, XS190-157, XS190-158, XS190-170, XS190-181, XS190- 182, XS190-183, XS190-186, XS197-3, XS197-4, XS197-5, XS197-6, XS197-14, XS197-29, XS197-38, XS197-49, XS197-50, XS197-70, XS197-71, XS197-72, XS197-73, XS197-74, XS197-85, XS197-93, XS197-94, XS197-96, XS197-133, XS197-145, XS197-176, XS197-185, XS197-186, XS209-21 , XS209-22, XS209-23, XS209-24, XS209-25, XS209-26, XS209-27, XS209-28, XS209-30, XS209-38, XS209-39, XS209-40, XS209-53, XS209-54, XS209-55, XS209-57, XS209-60, XS209-62, XS209-65, XS209-74, XS209-75, XS209-92, XS209-99, XS209-100, XS209-101, XS209-122, XS209-139, XS209-140, XS209-165, XS209-166, XS209- 167, XS209-168, XS209-174, XS209-175, XS209-176, XS209-184, XS209-185, XS224-6, XS224-106, XS224-107, XS224-108, XS224-109, XS224-110, XS224-111, XS224-116, XS224- 117, XS224-118, XS224-119, XS224-143, XS224-144, XS224-145, XS224-147, XS224-148, XS224-149, XS224-150, XS224-154, XS224-155, XS224-156, XS224-157, XS224-158, XS224- 159, or analogs thereof.

[0111] In some embodiments, the small molecules that covalently modify OTUB1 are

[0112] B. OTUBl-recruiting DUBTACs

[0113] The ubiquitination and degradation of many proteins is the cause of several classes of diseases. Therefore, de-ubiquitination and stabilization of these kinds of proteins may provide a novel therapeutic strategy.

[0114] In an aspect, this disclosure provides a method of treating AMPK, cGAS, or AF508-CFTR mediated diseases, the method including administering one or more AMPK, cGAS, or CFTR DUBTACs to a subject who has an AMPK, cGAS, or AF508-CFTR -mediated disease, the AMPK, cGAS, or CFTR DUBTACs being bivalent compounds including an AMPK, cGAS, or CFTR ligand conjugated to a de-ubiquitination tag via a linker, which would stabilize AMPK, cGAS, or CFTR. The AMPK, cGAS, or AF508-CFTR -mediated disease can be a disease resulting from AMPK, cGAS, or CFTR destabilization. The AMPK, cGAS, or AF508-CFTR-mediated disease can have reduced AMPK, cGAS, or CFTR expression relative to a wild-type tissue of the same species and tissue type.

[0115] More specifically, the present disclosure provides a bivalent compound including an AMPK, cGAS, or CFTR ligand conjugated to OTUB1 recruiter via a linker.

[0116] In some aspects, the AMPK, cGAS, or CFTR DUBTACs have the form “PI-Linker- OTUB1 Recruiter”, as shown below:

[0117] Formula (B) wherein PI (a ligand for a “protein of interest,” i.e., the protein to be subjected to de-ubiquitination) (e.g., a AMPK, cGAS, or CFTR ligand), and OTUB1 Recruiter comprises a de-ubiquitination tag (e g., a ligand for OTUB1). Exemplary AMPK, cGAS, or CFFTR ligands (PI), exemplary OTUB1 binders (OTUB1 Recruiters), and exemplary linkers (Linker) are illustrated below:

[0118] AMPK ligand

[0119] In some embodiments, the AMPK ligand can be an AMPK activator or AMPK inhibitor which potently binds to AMPK. In some embodiments, the AMPK ligand comprises Metformin (Xiao et al., 2020), AICAR (Nakamaru et al., 2005), ZMP, AMP, A-769662 (Sanders et al., 2007), 991 (Ngoei et al., 2018), PF-06685249 (Edmonds et al., 2018), PXL770 (Cusi et al., 2021), MK- 8722 (Myers et al., 2017), PF-739 (Aledavood et al., 2021), 4-azaindole, RSVA405 (Vingtdeux et al., 2011), COH-SR4 (Figarola and Rahbar, 2013), B10 (Sun et al., 2020), Dorsomorphin (Kim et al., 2011), BAY-3827 (Lemos et al., 2021), GSK-690693 (Rhodes et al., 2008) or a derivative thereof. In some embodiments, the AMPK ligand is a compound disclosed in one or more of WO2009124636, W02009100130, W02010036613, WO2011029855, WO2011080277, WO2011032320, WO2013116491, W02014133008, W02016008404, W02016001224, each of which is incorporated by reference in its entirety.

[0120] In some embodiments, the AMPK ligand is an AMPK inhibitor. In some embodiments, the AMPK ligand is an ATP-competitive AMPK inhibitor. In some embodiments, the AMPK ligand is an AMPK activator. In some embodiments, the AMPK ligand is an AMPK activator, where the AMPK activator binds to an allosteric binding site. In some embodiments, the AMPK activator binds to an allosteric binding site located at the α / β subunit interface. In some embodiments, the

[0121] AMPK activator has different AMPK isoform selectivity. In some embodiments, the AMPK activator is a pan-AMPK isoform activator.

[0122] In some embodiments, the AMPK ligand comprises the structure of Formula (B-I):

[0123] Formula (B-I), or a pharmaceutically acceptable salt of the compound or the tautomer thereof, wherein

[0124] AA is selected from

[0125] BAis selected from denotes the point of attachment to Linker in Formula (B);

[0126] CA is selected from N or CRA4;

[0127] Ring DA and Ring EAare independently selected from null, C3-C12cycloalkyl, 3-12- membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl;

[0128] XA and YA are independently selected from a bond, -O-, -S-, -NRA9-, -C(O)-, -C(O)O-, - C(O)NRA9-, -O-C(O)NRA9-, -NRA10C(O)NRA9-, -S(O)-, -S(O)2-, -S(O)NRA9-, -S(O)2NRA9-, C1- C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, and C2-C6alkynylene; RA1, RA2, each RA4, each RA5, and each RA7are independently selected from H, halogen, cyano, ORA11, NRAHRA12, C(O)ORAU, C(O)NRAURA12, S(O)2NRAURA12, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic; RA3is selected from H, halogen, cyano, ORA11, NRAHRA12, C(O)ORAU, C(O)NRAHRA12, S(O)2NRAHRA12, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, 3-12 membered heterocyclic, Ce-Cio aryl, and 5-10 membered heteroaryl; or RA1and RA2, RA2and RA3, RA3and RA4, two RA5, or two RA7, together with the atoms to which they are attached, optionally form C4-C6 cycloalkyl or 5-7 membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl; RA6and RA8are independently selected from a bond, -O-, -NRA13-, -C(O)-, -C(O)O-, - C(O)NRA13-, -O-C(O)NRA13-, -NRA14C(O)NRA13-, -S(O)-, -S(O)2-, -S(O)NRA13-, -S(O)2NRA13-, C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, C2-C6alkynylene, C6-C10arylene, and 5-10 membered heteroarylene; each RA9, each RA10, each RA11, each RA12, each RA13, and each RA14are independently selected from H, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic, Ce-Cio aryl, and 5-10 membered heteroaryl; or RA11and RA12together with the atoms to which they are attached, optionally form 3-12 membered heterocyclic; and mA and nA are independently selected from 0, 1, 2, 3, and 4.

[0129] In some embodiments, AA is N; and BA is . In some embodiments, AA and BA is N. In some embodiments,

[0130] In some embodiments, the AMPK ligand comprises the structure of Formulae (B-I-a) and Formulae (B-I-b) :

[0131]

[0132] Formula (B-I-a), and Formula (B-I-b), wherein

[0133] AA and are BA independently selected from N, CRA4

[0134] In some embodiments, XA is selected from -O-, -S-, -NRA9-, -C(O)-, C1-C6alkylene, C1- C6, haloalkylene, C1-C6heteroalkylene, C6-C10arylene, and 5-10 membered heteroarylene. In some embodiments, XA is selected from -O-, -S-, -NRA9-, -C(O)-, methylene, and halomethylene, In some embodiments, XAis selected from -O-, -S-, -NH-, -C(O)-, CH2, and CF2, In some embodiments, XA is -O-.

[0135] In some embodiments, YA is selected from a bond, -C(O)-, -C(O)O-, -C(O)NRA9-, -S(O)-, -S(O)2-, -S(O)NRA9-, -S(O)2NRA9-. In some embodiments, YA is selected from a bond, -C(O)-, - C(O)NRA9-, and -S(O)2NRA9-. In some embodiments, YA is -C(O)NRA9-.

[0136] In some embodiments, the AMPK ligand comprise the structure of Formulae (B-I-al), (B- I-bl), and (B-I-b2):

[0137] Formula (B-I-al), Formula (B-I-b 1), and Formula (B-I-b2).

[0138] In some embodiments, AA is N. In some embodiments, AA is CRA4. In some embodiments, BA is N. In some embodiments, BA is CRA4. In some embodiments, CA is N. In some embodiments, CA is CRA4. In some embodiments, Ring DAis selected from null, C3-C12cycloalkyl, 3-12-membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl. In some embodiments, Ring DAis C3- C12cycloalkyl. In some embodiments, Ring DAis selected from cyclobutyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. In some embodiments, Ring DA is 3-12- membered heterocyclic. In some embodiments, Ring DA is selected from tetrahydrofuryl, hexahydrofuro[3,2-Z>]furyl, azetidinyl, pyrrolidinyl, piperidinyl, and piperazinyl. In some embodiments, Ring DA is C6-C10aryl. In some embodiments, Ring DA is phenyl. In some embodiments, Ring DA is 5-10 membered heteroaryl. In some embodiments, Ring DA is selected from thiophenyl, benzothiophenyl, tetrahydrobenzothiophenyl, thiazolyl, imidazolyl, furanyl, pyrrolyl, pyrazolyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl, benzofuranyl, indolyl, and indazolyl.

[0139] In some embodiments, Ring EA selected from null, C3-C12cycloalkyl, 3-12-membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl. In some embodiments, Ring EA selected from nullC,6-C10aryl, and 5-10 membered heteroaryl. In some embodiments, Ring EA selected from null. In some embodiments, Ring EAISC6-C10aryl. In some embodiments, Ring EAis phenyl. . In some embodiments, Ring EA IS 5-10 membered heteroaryl. In some embodiments, Ring EA IS selected from thiophenyl, benzothiophenyl, tetrahydrobenzothiophenyl, thiazolyl, imidazolyl, furanyl, pyrrolyl, pyrazolyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl, benzofuranyl, indolyl, and indazolyl.

[0140] In some embodiments, the AMPK ligand comprises the structure of Formulae (B-I-a2), (B- I-a3), (B-I-a4), (B-I-b3), and (B-I-b4):

[0141] Formula (B-I-a2), Formula (B-I-a3), Formula (B-I-a4),

[0142]

[0143] Formula (B-I-b3), Formula (B-I-b4), wherein

[0144] FA is selected from N or CRA3; and

[0145] GA is selected from N or CRA7

[0146] In some embodiments, RA3is selected from C3-C12cycloalkyl, 3-12 membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl. In some embodiments, RA3is selected from C6-C10aryl and 5-10 membered heteroaryl.

[0147] In some embodiments, the AMPK ligand comprises the structure of Formulae (B-I-a5), (B- I-a6), (B-I-a7), (B-I-b5), and (B-I-b6):

[0148] Formula (B-I-b5), Formula (B-I-b6), wherein

[0149] ArA is selected from C6-C10aryl, or 5-10 membered heteroaryl; each RA15is independently selected from H, halogen, cyano, ORA11, NRAHRA12, C(O)ORA11C(O)NRA11R1A12, S(O)2NRA11RA12, -N=S(O)RA11RA12, CI-C6alkyl, haClo1a-Clk6yl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, 3-12 membered heterocyclic, C6-C10aryl and 5-10 membered heteroaryl, or two of RA15, together with the atoms to which they are attached, optionally form C3-C12cycloalkyl, 3-12 membered heterocyclic, C6-C10aryl or 5-10 membered heteroaryl; and

[0150] OA is selected from 0, 1, 2, 3, 4, 5, or 6.

[0151] In some embodiments, ArA is selected from C6-C1a0ryl and 5-10 membered heteroaryl. In some embodiments, ArA is selected from phenyl, naphthalene, indane, 5,6, 7,8- tetrahydronaphthalene, biphenyl, pyrazole, pyridine, pyrazine, pyrimidine, thiazole, thiophene, benzoimidazole, quinoline, isoquinoline, indole, indazole, carbazole, benzotriazole, benzofuran, benzothiazole, benzo[b]thiophene, benzo[d]isooxazole, 3,4-dihydro-2H-benzo[l,4]oxazine, benzo[l,3]dioxole, benzo[ 1,4] di oxane, l / / -pyrrolo[2,3-b]pyridine, [l,2,4]triazolo[4,3-a] pyridine, 3,4 dihydropyrido[3,2-b][l,4]oxazine, 3,4-dihydro-2H-l,4-benzoxazine, 2,3-dihydro-lH-indole, 2,3-dihydro-lH-isoindole, 2,3 -dihydrobenzoimidazole, 1,2-dihydroquinoline, 1, 2,3,4- tetrahydroisoqumoline, l,2,3,4-tetrahydrocyclopenta[b] indole, 1,2,3,4-tetrahydroquinoxaHne, 1,2,3,6-tetrahydropyridine. In some embodiments, ArA is selected from phenyl, pyridine, biphenyl, and indole. In some embodiments, ArA is indole.

[0152] In some embodiments, RA6is selected from a bond, -O-, -NRA13-, -C(O)-, -C(O)O-, - C(O)NRA13-, -S(O)2NRA13-, C1-C6alkylene, C1-C6haloalkylene, and C1-C6heteroalkylene. In some embodiments, RA6is selected from -C(O)-, -C(3-. In some embodiments, RA6is -C(O)NRA13-. In some embodiments, the N atom of the amide attaches two Linker-OTUBl Recruiter moieties.

[0153] In some embodiments, RA8is selected from a bond, -O-, -NRA13-, -C(O)-, -C(O)O-, - C(O)NRA13-, -S(O)2NRA13-, C1-C6alkylene, C1-C6haloalkylene, and C1-C6heteroalkylene.

[0154] In some embodiments, RA1is selected from H, halogen, cyano, C1-C6alkyl, C1-C6haloalkyl, Ci-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic. In some embodiments, RA1is selected from H and CH3. In some embodiments, RA1is H. In some embodiments, RA1is CH3.

[0155] In some embodiments, RA2is selected from H, halogen, cyano, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic. In some embodiments, RA2 is selected from halogen. In some embodiments, RA2selected from F or Cl. In some embodiments, RA2is F. In some embodiments, RA2is Cl.

[0156] In some embodiments, each RA4is selected from H, halogen, cyano, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic. In some embodiments, RA4is H.

[0157] In some embodiments, each RA5is selected from H, halogen, cyano, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic. In some embodiments, each RA5is selected from H, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl. In some embodiments, RA5is H, CH3, CH2CH3, and CH2OH.

[0158] In some embodiments, each RA7is selected from H, halogen, cyano, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic. In some embodiments, each RA7is selected from H, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl. In some embodiments, RA7is H, CH3, and CH2CH3.

[0159] In some embodiments, each RA9is independently selected from H, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic, C6- aCry1l0, and 5-10 membered heteroaryl. In some embodiments, each RA9is independently selected from H, Ci- C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic. In some embodiments, RA9is H.

[0160] In some embodiments, each RA10is independently selected from H, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic, C6- aCry10l, and 5-10 membered heteroaryl. In some embodiments, each RA10is independently selected from H, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic. In some embodiments, RA10is H.

[0161] In some embodiments, each RA11and each RA12are independently selected from H, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic, C6- C10aryl, and 5-10 membered heteroaryl. In some embodiments, each RA11and each RA12are independently selected from H, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic. In some embodiments, RA11is selected from H and CH3. In some embodiments, RA12is selected from H and CH3.

[0162] In some embodiments, RA11and RA12together with the atoms to which they are attached, optionally form 3-12 membered heterocyclic. In some embodiments, each RA13is independently selected from H, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic, C6- aCry1l0, and 5-10 membered heteroaryl. In some embodiments, each RA13is independently selected from H, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic.

[0163] In some embodiments, each RA14is independently selected from H, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic, C6- aCry1l0, and 5-10 membered heteroaryl. In some embodiments, each RA14is independently selected from H, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic.

[0164] In some embodiments, each RA15is independently selected from each RA15is independently selected from H, halogen, cyano, ORA11, NRA11A12, C(O)ORA11, C(O)NRA11RA12, S(O)2NRA' 'RA12, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, 3-12 membered heterocyclic, C6-C1a0ryl and 5-10 membered heteroaryl.

[0165] In some embodiments, each RA15is independently selected from each RA15is independently selected from H, halogen, cyano, ORA11, NRA’ 'RA12, S(O)2NRA1 1RA12, -N=S(O)RA11RA12, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, 3-12 membered heterocyclic, C6-C10aryl and 5-10 membered heteroaryl. In some embodiments, each RA15is independently selected from each RA15is independently selected from H, CN, CH3, cyclopropyl, hydroxymethyl cyclopropyl, OCH3, dimethyl amino, -N=S(O)(CH3)2, and -Ph-N=S(O)(CH3)2.

[0166] In some embodiments, mA is selected from 0, 1, 2, 3, and 4. In some embodiments, mA is selected from 0, 1, and 2. In some embodiments, nA is selected from 0, 1, 2, 3, and 4. In some embodiments, nA is selected from 0, 1, and 2. In some embodiments, OA is selected from 0, 1, 2, 3, 4, and 5.

[0167] In some embodiments, the AMPK ligand comprise the structure of Formulae (B-I), (B-I- a), (B-I-al), (B-I-a2) and (B-I-a5).

[0168] In some embodiments, the AMPK ligand comprise a derivative of following compounds:

[0169] Wherein, ♦* denotes the point of attachment to Linker. cGAS ligand

[0170] In some embodiments, the cGAS ligand is seleted from a cGAS activator or cGAS inhibitor which potently binds with cGAS. In some embodiments, the cGAS ligand is cGAS activator. In some embodiments, the cGAS ligand is cGAS inhibitor. In some embodiments, the cGAS ligand comprises hydroxychloroquinine (HCQ), Quinacrine (QC), X6, RU114757, RU191752, RU100840, RU.365, RU.521, J001, G001, G108, G150, GMO, CPD-25, aspirin, CPD-C, PF- 06928215(Zhao et al., 2022). In some embodiments, the cGAS ligand is a compound disclosed in one or more of U.S. 62 / 318,435, U.S. 62 / 355,403, U.S. 2018 / 0230115, WO2019241787,

[0171] W02019153002, WO2022140387, W02023004437, each of which is incorporated herein by reference in its entirety.

[0172] In some embodiments, the cGAS ligand comprise the structure of Formula (C-I):

[0173] Formula (C-I), or a pharmaceutically acceptable salt of the compound or the tautomer thereof, wherein denotes the point of attachment to Linker in Formula (B);

[0174] ♦ ♦ connects to either RBlbor RB311, with the proviso that when ** connects to RBlb,

[0175] RB3is RB3a; and when ** connects to Rs3b, RB1is RB1S;

[0176] AB is selected from N and CRB4;

[0177] BB is selected from N, O, S, CRB5and NRB5;

[0178] CB is selected from N and C; DB is selected from N and C;

[0179] XBis XBa-XBb;

[0180] XBais selected from a bond, -C(O)-, -C(O)O-, -C(O)NRB6-, -S(O)-, -S(O)2-, -S(O)NRB6-, - S(O)2NRB6-;

[0181] XBbis selected from a bond, C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C3- C12 cycloalkylene, and 3-12 membered heterocyclicene, C6-C10arylene, and 5-10 membered heteroarylene; RB1is selected from monovalent group RB1aand bivalent group RBlb;

[0182] RBlais selected from H, halogen, cyano, ORB7, NRB7RB8, C(O)ORB7, C(O)NRB7RB8, S(O)2NRB7RB8, C1-C6alkyl, C1-C ha6loalkyl, C1- hCet6eroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3- C12cycloalkyl, 3-12 membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl;

[0183] RBlbis connected to the Linker, and is selected from a bond, -O-, -NRB7-, -C(O)O-, - C(O)NRB7-, -S(O)2NRB7-, C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, C2-C6alkynylene, C3-C12cycloalkylene, 3-12 membered heterocyclicene, C6-C10arylene, and 5-10 membered heteroarylene; each RB2is independently selected from H, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, or two RB2, together with the atoms to which they are attached, optionally form C3-C12cycloalkyl or 3-12 membered heterocyclic; RB3is selected from monovalent group RB3aand bivalent group RB3b;

[0184] RB3ais selected from H, halogen, ORB9, NRB9RB10, C(O)ORB9, C(O)NRB9RB10, S(O)2NRB9RB10, C1-C6alkyl, C1- hCal6oalkyl, C h1-eCte6roalkyl, C2-C6alkenyl, C2-C6alkynyl, C3- Ci2cycloalkyl, and 3-12 membered heterocyclic;

[0185] RB3bis connected to the Linker, and is selected from a bond, -O-, -NRB9-, -C(O)O-, - C(O)NRB9-, -S(O)2NRB9-, C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, C2-C6alkynylene, C3-Ci2cycloalkylene, and 3-12 membered heterocyclicene; each RB4is independently selected from H, halogen, cyano, ORB7, NRB7RB8, C(O)ORB7, C(O)NRB7RB8, S(O)2NRB7RB8, C1-C3alkyl, C1-C3haloalkyl, C1-C3heteroalkyl, C2-C4 alkenyl, C2- C4 alkynyl, C3-C6 cycloalkyl, and 3-6 membered heterocyclic, or two RB4, together with the atoms to which they are attached, optionally form partially unsaturated C3-C12cycloalkyl, partially unsaturated 3-12 membered heterocyclic, C6-C ar10ylene, or 5-10 membered heteroarylene; RB5is selected from H, halogen, C1-C3alkyl, C1-C3haloalkyl, C1-C3heteroalkyl, C3-C6cycloalkyl, and 3-6 membered heterocyclic; RB6, each RB7, each RB8, each RB9, and each RB10, are independently selected from H, C1- C6 alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic, or RB7and RB8, and RB9and RB10together with the atoms to which they are attached, optionally form 3-12 membered heterocyclic; mB is selected from 1 and 2; riB is selected from 0, 1, 2 and 3; and

[0186] OB is selected from 0, 1, 2, 3, 4, 5 and 6.

[0187] In some embodiments, * connects to RB1b, RB3is RB3a. In some embodiments, ** connects to RB3b, RB1is RB1 1

[0188] In some embodiments, the cGAS ligand comprises the structures of Formulae (C-I-a) and (C-I-b):

[0189] Formula (C-I-a), Formula (C-I-b).

[0190] In some embodiments, AB is CRB4In some embodiments, BB is NRB5. In some embodiments, CB is C. In some embodiments, DB is C.

[0191] In some embodiments, the cGAS ligand comprises the structures of Formulae (C-I-al) and (C-I-bl):

[0192]

[0193] Formula (C-I-al), Formula (C-I-bl).

[0194] In some embodiments, RB1ais selected from H, halogen, C3-C12cycloalkyl, 3-12 membered heterocyclic,C6-C10aryl, and 5-10 membered heteroaryl. In some embodiments, RB13is selected from C3-C12cycloalkyl, 3-12 membered heterocyclic, C6-C a1r0yl, and 5-10 membered heteroaryl. In some embodiments, RB1ais selected from C6-C ar10yl, and 5-10 membered heteroaryl. In some embodiments, RB1ais 5-10 membered heteroaryl. In some embodiments, RB13is selected from furyl, thiophenyl, pyrrolyl, pyrazolyl, oxazolyl, oxadiazolyl, thiazolyl, isoxazolyl, isothiazolyl, imidazolyl, triazolyl, pyridinyl, pyrimidinyl, pyrazinyl, and pyridazinyl. In some embodiments, RB13is pyrazolyl. In some embodiments, RB11is = pyridinyl.

[0195] In some embodiments, RBlbis selected from C3-C12cycloalkylene, 3-12 membered heterocyclicene, C6-C10arylene, and 5-10 membered heteroarylene. In some embodiments, RB11is selected from C6-C10arylene, and 5-10 membered heteroarylene.

[0196] In some embodiments, the cGAS ligand comprises the structures of Formulae (C-I-a2) and (C-I-b2):

[0197] Formula (C-I-a2), Formula (C-I-b2), wherein

[0198] ArB is selected from C6-C10aryl, and 5-10 membered heteroaryl; each RB1 1is independently selected from H, halogen, cyano, ORB7, NRB7RB8, C(O)ORB7, C(O)NRB7RB8, S(O)2NRB7RB8, C1-C6alkyl, C h1a-lCo6alkyl, hCe1t-eCr6oalkyl, C2-C6alkenyl, C2- C6, alkynyl, C3-C12cycloalkyl, 3-12 membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl; or two RB11, together with the atoms to which they are attached, optionally form partially unsaturated C3-C12cycloalkyl, partially unsaturated 3-12 membered heterocyclic, C6-C a1r0yl, or 5-10 membered heteroaryl;

[0199] PB is selected from 0, 1, 2, 3, 4, and 5; and

[0200] PB-I is selected from 0, 1, 2, 3, and 4.

[0201] In some embodiments, XB3is selected from -C(O)-, -S(O)-, and -S(O)2-. In some embodiments, XBais -C(O)-. In some embodiments, XBbis selected from C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C3-C12cycloalkylene, and 3-12 membered heterocyclicene.

[0202] In some embodiments, the cGAS ligand comprises the structures of Formulae (C-I-a3) and (C-I-b3):

[0203] Formula (C-I-a3), Formula (C-I-b3), wherein each RB12is independently selected from H, halogen, ORB7, NRB7RB8, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic; or two RB12, RB12and RB3a, and RB12and RBb, together with the atoms to which they are attached, optionally form C3-C12cycloalkyl, 3-12 membered heterocyclic; and qB is sleeted from 0, 1, 2, 3, 4, and 5.

[0204] In some embodiments, ArB is 5-10 membered heteroaryl. In some embodiments, ArB is selected from furyl, thiophenyl, pyrrolyl, pyrazolyl, oxazolyl, oxadiazolyl, thiazolyl, isoxazolyl, isothiazolyl, imidazolyl, triazolyl, pyridinyl, pyrimidinyl, pyrazinyl, and pyridazinyl. In some embodiments, ArB is pyrazolyl. In some embodiments, ArB is pyridinyl.

[0205] In some embodiments, the cGAS ligand comprises the structures of Formulae (C-I-a4) and (C-I-b4):

[0206]

[0207] Formula (C-I-a4), Formula (C-I-b4).

[0208] In some embodiments, each RB2is independently selected from H, C1-C3alkyl, C1-C3haloalkyl, C1-C3heteroalkyl, C3-C6cycloalkyl. In some embodiments, RB2is H. In some embodiments, RB2is C1-C3alkyl. In some embodiments, RB2is CH3. In some embodiments, two RB2, together with the atoms to which they are attached, optionally form C3-C6cycloalkyl or 3-6 membered heterocyclic. In some embodiments, two RB2, together with the atoms to which they are attached, optionally form C3-C5 cycloalkyl.

[0209] In some embodiments, Ru3ais selected from H, halogen, ORB9, NRB9RB10, C1-C6alkyl, Ci- Cg haloalkyl, Ci-Cg heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic. In some embodiments, RB3ais selected from H, ORB9and NRB9RB10. In some embodiments, RB3ais selected from H, OH, OCH3, OCH2CH2OCH3, NH2, NHCH3, and NHC(O)CH2NH2. In some embodiments, RB3ais OH.

[0210] In some embodiments, RB3bis selected from a bond, -O-, -NRB9-, Ci-Cg alkylene, C1-C6haloalkylene, Ci-Cg heteroalkylene, C3-C12cycloalkylene, and 3-12 membered heterocyclicene. In some embodiments, RB31)is selected from a bond, -O-, -NRB9-. In some embodiments, RB3bis selected from a bond, -O-, -NH-, and -N(CH3)-. In some embodiments, RB3bis -O-.

[0211] In some embodiments, each RB4is independently selected from H, halogen, cyano, ORB7, NRB7RB8, C1-C3alkyl, C1-C3haloalkyl, C1-C3heteroalkyl, C2-C4alkenyl, C2-C4alkynyl, C3-C6cycloalkyl, and 3-6 membered heterocyclic. In some embodiments, each RB4is independently selected from H. In some embodiments, each RB4is independently selected from cyano. In some embodiments, each RB4is independently selected from halogen. In some embodiments, each RB4is independently selected from F. In some embodiments, each RB4is independently selected from Cl. In some embodiments, each RB4is independently selected from methyl, ethyl, propyl, isopropyl, cyclopropyl, trifluoromethyl, difluoromethyl, methoxy, ethoxy, cyclopropoxy, and azetidinyl. In some embodiments, each RB4is independently selected from H. In some embodiments, each RB4 is independently selected from cyano. In some embodiments, each RB4is independently selected from halogen. In some embodiments, each RB4is independently selected from F. In some embodiments, each RB4is independently selected from Cl. In some embodiments, each RB4is independently selected from Br.

[0212] In some embodiments, two RB4, together with the atoms to which they are attached, optionally form partially unsaturated C3-C6cycloalkyl, partially unsaturated 3-6 membered heterocyclic,C6-C10aryl, or 5-10 membered heteroaryl.

[0213] In some embodiments, RB5is selected from H, C1-C3alkyl, C1-C3haloalkyl, C1-C3heteroalkyl, C3-C6cycloalkyl, and 3-6 membered heterocyclic. In some embodiments, RB5is selected from H and C1-C3alkyl. In some embodiments, RB5is H. In some embodiments, RB5is CH3.

[0214] In some embodiments, RB6, each RB7, each RB8, each RB9, and each RB10, are independently selected from H, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic. In some embodiments, RB6, each RB7, each RB8, each RB9, and each RB10, are independently selected from H, methyl, ethyl, propyl, isopropyl, cyclopropyl, trifluoromethyl, and difluoromethyl. In some embodiments, RB6, each RB7, each RB8, each RB9, and each RB10, are H. In some embodiments, RB6, each RB7, each RB8, each RB9, and each RB10, are methyl.

[0215] In some embodiments, RB7and RB8, and RB9and RB10together with the atoms to which they are attached, optionally form 3-6 membered heterocyclic.

[0216] In some embodiments, each RB11is independently selected from H, halogen, cyano, ORB7, NRB7RB8, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, 3-12 membered heterocyclic. In some embodiments, RB11is H. In some embodiments, each RB11is CH3.

[0217] In some embodiments, two RB11, together with the atoms to which they are attached, optionally form partially unsaturated C3-C12cycloalkyl, 3-12 membered partially unsaturated heterocyclic,C6-C10aryl, or 5-10 membered heteroaryl.

[0218] In some embodiments, each RB12is independently selected from H, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic. In some embodiments, each RB12is independently selected from H, and C1-C6alkyl. In some embodiments, RB12is H. In some embodiments, two RB12, RB12and RB3a, and RB12and RB3b, together with the atoms to which they are attached, optionally form C3-C6cycloalkyl, 3-6 membered heterocyclic.

[0219] In some embodiments, mBis 1. In some embodiments, nB is 1. In some embodiments, nB is 2. In some embodiments, OB is 0. In some embodiments, OB is 1. In some embodiments, OB is 2. In some embodiments, PB is 0. In some embodiments, PB is 1. In some embodiments, PB is 2. In some embodiments, PB-I is 0. In some embodiments, PB-I is 1. In some embodiments, qB is 0. In some embodiments, qβ is 1. In some embodiments, qβ is 2.

[0220] In some embodiments, the cGAS ligand comprises the structures of Formulae (C-I), (C-I- a), (C-I-al), (C-I-a2), (C-I-a3), and (C-I-a4).

[0221] In some embodiments, the cGAS ligand may be a derivative of following compounds:

[0222] Wherein, ♦* denotes the point of attachment to Linker.

[0223] CFTR Ligand

[0224] In some embodiments, the CFTR ligand can be a CFTR potentiator which potently binds with either wild type CFTR or mutant CFTR. In some embodiments, the CFTR ligand comprises ivacftor, lumacaftor, tezacaftor, elexacafor, or icenticaftor, or derivatives thereof. In some embodiments, the CFTR ligand is a compound disclosed in one or more of U.S. Patent No. 7,999,113; U.S. Patent No. 8,247,436; U.S. 8,410,274; WO 2011 / 133953; and WO2018 / 037350, each of which is incorporated by reference in its entirety.

[0225] In some embodiments, the CFTR ligand comprises the structures of Formula (D-I):

[0226] Formula (D-I), or a tautomer thereof, or a pharmaceutically acceptable salt of the compound or the tautomer, wherein denotes the point of attachment to Linker in Formula (B).

[0227] Ac and Cc are independently selected from O, S, or C(Rc8)(Rc9);

[0228] Bc is C(Rc10)(Rc11) or NRc12;

[0229] Arc1and Arc2are independently selected from null, C6-C10aryl and 5-10 membered heteroaryl;

[0230] Rc1and Rc2are independently selected from H, halo, ORc13, NRc13Rc14, C(O)ORc13, C(O)NRC13RC14, S(O)2NRC13RC14, C1-C6alkyl, hCal1o-Cal6kyl, heCte1-rCoa6lkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic, or

[0231] Rc1and Rc2, together with the atoms to which they are attached, optionally form C3-C12cycloalkyl; Rc3and Rc6are independently selected from H, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic; each Rc4, each Rc5, and each Rc7are independently selected from H, halogen, cyano, ORc13, NRc13Rc14, C(O)ORc13, C(O)NRC13RC14, S(O)2NRC13RC14, CI-C6alkyl, hCal1o-aClk6yl, Ci-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic, two Rc4, two Rc5, or two Rc7, together with the atoms to which they are attached, optionally form partially unsaturated C3-C12cycloalkyl, partially unsaturated 3-12 membered heterocyclic, C6-C10aryl and 5-10 membered heteroaryl;

[0232] Rc8, Rc9, Rc10and Rc11are independently selected from H, halogen, ORc13, NRc13Rc14, C(O)ORc13, C(O)NRc13Rc14, S(O)2NRC13RC14, CI-C6alkyl, C h1a-lCo6alkyl, C h1e-tCe6roalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic, or

[0233] Rc8and Rc9, and Rc10and Rc11, together with the atoms to which they are attached, optionally form C3-C12cycloalkyl and 3-12 membered heterocyclic;

[0234] Rc12is selected from H, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic;

[0235] Rc13and Rc14are independently selected from H, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic, or

[0236] Rc13and Rc14together with the atoms to which they are attached, optionally form 3-12 membered heterocyclic; me is 0, 1 , 2, or 3; nc is 0, 1, 2, or 3; and oc is 0, 1, 2, or 3.

[0237] In some embodiments, Arc1is 5-10 membered heteroaryl. In some embodiments, Arc1is selected from furyl, thiophenyl, pyrrolyl, pyrazolyl, oxazolyl, oxadiazolyl, thiazolyl, isoxazolyl, isothiazolyl, imidazolyl, triazolyl, pyridinyl, pyrimidinyl, pyrazinyl, and pyridazinyl. In some embodiments, ArB is pyridinyl. In some embodiments, Arc2is C6-C1a0ryl. In some embodiments, Arc2is phenyl.

[0238] In some embodiments, the CFTR ligand comprises the structures of Formula (D-I-a):

[0239] Formula (D-I-a), or a tautomer thereof, or a pharmaceutically acceptable salt of the compound or the tautomer.

[0240] In some embodiments, the CFTR ligand comprises the structures of Formula (D-I-al):

[0241] Formula (D-I-a 1), or a tautomer thereof, or a pharmaceutically acceptable salt of the compound or the tautomer.

[0242] In some embodiments, Ac is O. In some embodiments, Be is C(Rc10)(Rc11). In some embodiments, Cc is O.

[0243] In some embodiments, the CFTR ligand comprises the structures of Formula (D-I-a2):

[0244] Formula (D-I-a2), or a tautomer thereof, or a pharmaceutically acceptable salt of the compound or the tautomer.

[0245] In some embodiments, Rc1is selected from H, C1-C3alkyl, C2-C3alkenyl, C2-C3alkynyl, C3-C5 cycloalkyl, and 3-5 membered heterocyclic. In some embodiments, Rc2is selected from H, C1-C3alkyl, C2-C3 alkenyl, C2-C3 alkynyl, C3-C5 cycloalkyl, and 3-5 membered heterocyclic. In some embodiments, Rc1and Rc2, together with the atoms to which they are attached, optionally form C3-C6cycloalkyl. In some embodiments, Rc1and Rc2, together with the atoms to which they are attached, optionally form ]cyclopropyl. In some embodiments, Rc3is selected from H, C1-C6alkyl, C1-C6haloalkyl, and C1-C6heteroalkyl. In some embodiments, Rc3is H.

[0246] In some embodiments, each Rc4is independently selected from H, halo, cyano, ORc13, NRC13RC14, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic. In some embodiments, each Rc4is independetly selected from H and C1-C6alkyl. In some embodiments, each Rc4is independently selected from H and CH3.

[0247] In some embodiments, each Rc5is independently selected from H, halo, cyano, ORc13, NRC13RC14, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic. In some embodiments, each Rc3is independently selected from H and C1-C6alkyl. In some embodiments, Rc5is H.

[0248] In some embodiments, Rc6is selected from H, C1-C6alkyl, C1-C6haloalkyl, and C1-C6heteroalkyl. In some embodiments, Rc6is H. In some embodiments, Rc3is CH3.

[0249] In some embodiments, each Rc7is independently selected from H, halogen, cyano, ORc13, NRC13RC14, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic. In some embodiments, each Rc7is independently selected from H and C1-C6alkyl. In some embodiments, Rc7is H.

[0250] In some embodiments, Rc8and Rc9are independently selected from H, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic. In some embodiments, Rc8and Rc9are independently selected from H and C1-C6alkyl,

[0251] In some embodiments, Rc10and Rc11are independently selected from H, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic. In some embodiments, Rc10and Rc11are independently selected from H, halogen and C1-C6alkyl. In some embodiments. Rc10and Rc11are independently selected from H, F, Cl, CH3, and CF3. In some embodiments, both Rc10and Rc11are F.

[0252] In some embodiments, Rc12is selected from H, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic.

[0253] In some embodiments, Rc13and Rc14are independently selected from H, C1-C6alkyl, Ci- C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclic. In some embodiments, Rc13and Rc14are independently selected from H and C1-C6alkyl. In some embodiments, Rc13and Rc14are independently selected from H and CH3. In some embodiments, Rc13and Rc14together with the atoms to which they are attached, optionally form 3-6 membered heterocyclic.

[0254] In some embodiments, me is 1. In some embodiments, nc is 0. In some embodiments, oc is 0.

[0255] In some embodiments, the CFTR ligand comprises the structures of Formulae (D-I-a3) and (D-I-a4):

[0256] Formula (D-I-a3), Formula (D-I-a4).

[0257] In some embodiments, the CFTR ligand may be a derivative of following compounds:

[0258] Wherein, •* denotes the point of attachment to Linker.

[0259] Linkers

[0260] In all of the above-described compounds, the AMPK, cGAS or CFTR ligand is conjugated to the de-ubiquitination tag through a linker. The linker can include, for example, acyclic or cyclic saturated or unsaturated carbon, ethylene glycol, amide, amino, ether, urea, carbamate, aromatic, heteroaromatic, heterocyclic and / or carbonyl containing groups with different lengths. In some embodiments, the linker can be a moiety of Formula (E):

[0261] Formula (E) wherein

[0262] A, W and B, at each occurrence, are independently selected from null, or bivalent moiety and R NR1S(O)2N(R2)R , wherein

[0263] R and R are independently selected from a bond, optionally substituted Rr-(C1-C8alkyl), or a moiety comprising of optionally substituted C1-C8alkyl, optionally substituted C2-C8alkenyl, optionally substituted C2-C8alkynyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkoxyC1-C8alkyl, optionally substituted C1-C8alkylaminoC1-C8alkyl, optionally substitutedC1-C8haloalkyl, optionally substituted C1-C8alkylene, optionally substituted C2-C8alkenylene, optionally substituted C2-C8alkynylene, optionally substituted C1-C8hydroxy alkylene, optionally substituted C1-C8alkoxyC1-C8alkylene, optionally substituted C1-C8alkylaminoCi- Csalkylene, optionally substituted C1-C8haloalkylene, optionally substituted 3-10 membered carbocyclic, optionally substituted 4-10 membered heterocyclic, optionally substituted C3-C13fused cycloalkyl, optionally substituted C3-C13fused heterocyclic, optionally substituted C3-C13bridged cycloalkyl, optionally substituted C3-C13bridged heterocyclic, optionally substituted C3- C13 spiro cycloalkyl, optionally substituted C3-C13spiro heterocyclic, optionally substituted aryl, and optionally substituted heteroaryl;

[0264] Rris selected from optionally substituted 3-10 membered carbocyclic, optionally substituted 4-10 membered heterocyclic, optionally substituted C3-C13fused cycloalkyl, optionally substituted C3-C13fused heterocyclic, optionally substituted C3-C13bridged cycloalkyl, optionally substituted C3-C13bridged heterocyclic, optionally substituted C3-C13spiro cycloalkyl, optionally substituted C3-C13spiro heterocyclic, optionally substituted aryl, and optionally substituted heteroaryl; R1and R2are independently selected from hydrogen, optionally substituted C1-C8alkyl, optionally substituteCd2-C8alkenyl, optionally substituted C2-C8alkynyl, optionally substituted C1-C8alkoxyalkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkylaminoC1-C8alkyl, optionally substituted 3-10 membered carbocyclic, optionally substituted 4-10 membered heterocyclic, optionally substituted aryl, and optionally substituted heteroaryl;

[0265] R and R , R1and R2, R and R1, R andR2, R and R1, R andR2together with the atom to which they are connected form a 3-20 membered cycloalkyl or 4-20 membered heterocyclic ring; and m is 0 to 15.

[0266] In one embodiment, the linker moiety is of Formula (E-l)

[0267] Formula (E-l), wherein

[0268] R1, R2, R3and R4, at each occurrence, are independently selected from hydrogen, halogen, hydroxyl, amino, cyano, nitro, optionally substituted C1-C8alkyl, optionally substituteC2-C8alkenyl, optionally substituted C2-C8alkynyl, optionally substituted C1-C8alkoxy, optionally substituted C1-C8alkoxyalkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkylamino, and optionally substituted C1-C8alkylaminoC1-C8alkyl, optionally substituted 3-10 membered carbocyclic, optionally substituted 3-8 membered cycloalkoxy, optionally substituted 3-10 membered carbocyclicamino, optionally substituted 4-8 membered membered heterocyclic, optionally substituted aryl, and optionally substituted heteroaryl, or

[0269] R1and R2, R3and R4together with the atom to which they are connected form a 3-20 membered cycloalkyl or 4-20 membered heterocyclic ring; A, W and B, at each occurrence, are independently selected from null, or bivalent moiety selected from R -R ”, R COR , R CO2R , R C(O)N(R5)R , R’C(S)N(R5)R”, R OR”, R OC(O)R”, R OC(O)OR ”, R OCONR5R”, R SR ”, R SOR”, R SO2R , R SO2N(R5)R”, RN(R5)R”, RNR5COR ”, RNR5C(O)OR’ , RNR5CON(R6)R”, R NR5C(S)R ”, RNR5S(O)R ”, RNR5S(O)2R ”, and R NR5S(O)2N(R6)R ”, wherein

[0270] R and R are independently selected from null, optionally substituted Rr-(C1-C8alkyl), or a moiety comprising of optionally substituted C1-C8alkyl, optionally substituted C2-C8alkenyl, optionally substituted C2-C8alkynyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkoxyC1-C8alkyl, optionally substituted C1-C8alkylaminoC1-C8alkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1-C8alkylene, optionally substituted C2-C8alkenylene, optionally substituted C2-C8alkynylene, optionally substituted C1-C8hydroxy alkylene, optionally substituted C1-C8alkoxyC1-C8alkylene, optionally substituted C1-C8alkylaminoCi- Csalkylene, optionally substituted C1-C8haloalkylene, optionally substituted 3-10 membered carbocyclic, optionally substituted 4-10 membered heterocyclic, optionally substituted C3-C13fused cycloalkyl, optionally substituted C3-C13fused heterocyclic, optionally substituted C3-C13bridged cycloalkyl, optionally substituted C3-C13bridged heterocyclic, optionally substituted C3- C13 spiro cycloalkyl, optionally substituted C3-C13spiro heterocyclic, optionally substituted aryl, and optionally substituted heteroaryl;

[0271] Rris selected from optionally substituted 3-10 membered carbocyclic, optionally substituted 4-10 membered heterocyclic, optionally substituted C3-C13fused cycloalkyl, optionally substituted C3-C13fused heterocyclic, optionally substituted C3-C13bridged cycloalkyl, optionally substituted C3-C13bridged heterocyclic, optionally substituted C3-C13spiro cycloalkyl, optionally substituted C3-C13spiro heterocyclic, optionally substituted aryl, and optionally substituted heteroaryl;

[0272] R5and R6are independently selected from hydrogen, optionally substituted C1-C8alkyl, optionally substituted C2-C8alkenyl, optionally substituted C2-C8alkynyl, optionally substituted C1-C8alkoxyalkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkylaminoC1-C8alkyl, optionally substituted 3-10 membered carbocyclic, optionally substituted 4-10 membered heterocyclic, optionally substituted aryl, and optionally substituted heteroaryl; R and R , R5and R6, R and R5, R andR6, R and R5, R andR6together with the atom to which they are connected form a 3-20 membered cycloalkyl or 4-20 membered heterocyclic ring; m is 0 to 15; n, at each occurrence, is 0 to 15; and o is 0 to 15.

[0273] In another embodiment, the linker moiety is of Formula (E-2)

[0274] Formula (E-2), wherein

[0275] R1and R2, at each occurrence, are independently selected from hydrogen, halogen, hydroxyl, amino, cyano, nitro, and optionally substituted C1-C8alkyl, optionally substituted C1-C8alkoxy, optionally substituted C1-C8alkoxy C1-C8alkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkylamino, Ci- CsalkylaminoC1-C8alkyl, optionally substituted 3-10 membered carbocyclic, optionally substituted 3-8 membered cycloalkoxy, optionally substituted 3-10 membered carbocyclicamino, optionally substituted 4-10 membered heterocyclic, optionally substituted aryl, and optionally substituted heteroaryl, or

[0276] R1and R2together with the atom to which they are connected form a 3-20 membered cycloalkyl or 4-20 membered heterocyclic ring;

[0277] A and B, at each occurrence, are independently selected from null, or bivalent moiety selected from R -R , R COR , R CO2R , R C(O)NR3R’ , R C(S)NR3R”, R’OR”, R OC(O)R’ , R OC(O)OR , R OCON(R3)R , R SR , R SOR , R SO2R , R SO2N(R3)R , R N(R3)R , R’NR3COR”, RNR3C(O)OR ”, R’NR3CON(R4)R’, R’NR3C(S)R ”, R’NR3S(O)R”, R’NR3S(O)2R”, and R NR3S(O)2N(R4)R ”, wherein

[0278] R and R are independently selected from null, optionally substituted Rr-(C1-C8alkyl), or a moiety comprising of optionally substituted C1-C8alkyl, optionally substituted C2-C8alkenyl, optionally substitutedC2-C8alkynyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkoxyC1-C8alkyl, optionally substituted C1-C8alkylaminoC1-C8alkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1-C8alkylene, optionally substituted C2-C8 alkenylene, optionally substituted C2-C8alkynylene, optionally substituted C1-C8hydroxyalkylene, optionally substituted C1-C8alkoxyC1-C8alkylene, optionally substituted C1-C8alkylaminoCi- Csalkylene, optionally substituted C1-C8haloalkylene, optionally substituted 3-10 membered carbocyclic, optionally substituted 4-10 membered heterocyclic, optionally substituted C3-C13fused cycloalkyl, optionally substituted C3-C13fused heterocyclic, optionally substituted C3-C13bridged cycloalkyl, optionally substituted C3-C13bridged heterocyclic, optionally substituted C3- C13 spiro cycloalkyl, optionally substituted C3-C13spiro heterocyclic, optionally substituted aryl, and optionally substituted heteroaryl;

[0279] Rris selected from optionally substituted 3-10 membered carbocyclic, optionally substituted 4-10 membered heterocyclic, optionally substituted C3-C13fused cycloalkyl, optionally substituted C3-C13fused heterocyclic, optionally substituted C3-C13bridged cycloalkyl, optionally substituted C3-C13bridged heterocyclic, optionally substituted C3-C13spiro cycloalkyl, optionally substituted C3-C13spiro heterocyclic, optionally substituted aryl, and optionally substituted heteroaryl;

[0280] R3and R4are independently selected from hydrogen, optionally substituted C1-C8alkyl, optionally substituted C2-C8alkenyl, optionally substituted C2-C8alkynyl, optionally substituted C1-C8alkoxyalkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkylaminoC1-C8alkyl, optionally substituted 3-10 membered carbocyclic, optionally substituted 4-10 membered heterocyclic, optionally substituted aryl, and optionally substituted heteroaryl;

[0281] R and R , R3and R4, R and R3, R and R4, R and R3, R and R4together with the atom to which they are connected form a 3-20 membered cycloalkyl or 4-20 membered heterocyclic ring; m, at each occurrence, is 0 to 15; and n is 0 to 15.

[0282] In another embodiment, the linker moiety is of Formula (E-3):

[0283] Formula (E-3) wherein X is selected from O, NH, and NR7;

[0284] R1, R2, R3, R4, R5, and R6, at each occurrence, are independently selected from hydrogen, halogen, hydroxyl, amino, cyano, nitro, optionally substituted C1-C8alkyl, optionally substituted C2-C8 alkenyl, optionally substituted C2-C8alkynyl, optionally substituted C1-C8alkoxy, optionally substituted C1-C8alkoxy C1-C8alkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkylamino, optionally substituted C1-C8alkylaminoC1-C8alkyl, optionally substituted 3-10 membered carbocyclic, optionally substituted 3-8 membered cycloalkoxy, optionally substituted 4-10 membered heterocyclic, optionally substituted aryl, and optionally substituted heteroaryl;

[0285] A and B are independently selected from null, or bivalent moiety selected from R -R , R COR , R CO2R ”, R C(O)N(R8)R’ , R C(S)N(R8)R’ , R OR , R OC(O)R”, R OC(O)OR”, R OCON(R8)R , R SR , R SOR , R SO2R , R SO2N(R8)R , R N(R8)R , R NR8COR , R’NR8C(O)OR”, R’NR8CON(R9)R”, R’NR8C(S)R”, R’NR8S(O)R”, R’NR8S(O)2R”, and RNR8S(O)2N(R9)R ’, wherein

[0286] R and R are independently selected from null, optionally substituted Rr-(C1-C8alkyl), or a moiety comprising of optionally substituted C1-C8alkyl, optionally substituted C2-C8alkenyl, optionally substitutedC2-C8alkynyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkoxyC1-C8alkyl, optionally substituted C1-C8alkylaminoC1-C8alkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1-C8alkylene, optionally substituted C2-C8alkenylene, optionally substituted C2-C8alkynylene, optionally substituted C1-C8hydroxyalkylene, optionally substituted C1-C8alkoxyC1-C8alkylene, optionally substituted C1-C8alkylaminoCi- Csalkylene, optionally substituted C1-C8haloalkylene, optionally substituted 3-10 membered carbocyclic, optionally substituted 4-10 membered heterocyclic, optionally substituted C3-C13fused cycloalkyl, optionally substituted C3-C13fused heterocyclic, optionally substituted C3-C13bridged cycloalkyl, optionally substituted C3-C13bridged heterocyclic, optionally substituted C3- C13 spiro cycloalkyl, optionally substituted C3-C13spiro heterocyclic, optionally substituted aryl, and optionally substituted heteroaryl;

[0287] Rris selected from optionally substituted 3-10 membered carbocyclic, optionally substituted 4-10 membered heterocyclic, optionally substituted C3-C13fused cycloalkyl, optionally substituted C3-C13fused heterocyclic, optionally substituted C3-C13bridged cycloalkyl, optionally substituted C3-C13bridged heterocyclic, optionally substituted C3-C13spiro cycloalkyl, optionally substituted C3-C13spiro heterocyclic, optionally substituted aryl, and optionally substituted heteroaryl;

[0288] R7, R8and R9are independently selected from hydrogen, optionally substituted C1-C8alkyl, optionally substituted C2-C8alkenyl, optionally substituted C2-C8alkynyl, optionally substituted C1-C8alkoxyalkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkylaminoC1-C8alkyl, optionally substituted 3-10 membered carbocyclic, optionally substituted 4-10 membered heterocyclic, optionally substituted aryl, and optionally substituted heteroaryl;

[0289] R and R , R8and R9, R and R8, R and R9, R and R8, R and R9together with the atom to which they are connected form a 3-20 membered cycloalkyl or 4-20 membered heterocyclic ring; m, at each occurrence, is 0 to 15; n, at each occurrence, is 0 to 15; o is 0 to 15; and p is 0 to 15.

[0290] In some embodiments, in Formular (E-3), m and n is 0 or 1, and p is 0 to 15;

[0291] In some embodiments, in Formular (E-3), X is selected from O and NH;

[0292] In some embodiments, in Formular (E-3), R1, R2, R3, R4, R5, and R6, are independently selected from hydrogen, and optionally substituted C1-C6alkyl.

[0293] In some embodiments, Formulae E, E-l, E-2, and E-3, the linker moiety comprises a ring selected from the group consisting of a 3 to 13 membered ring, a 3 to 13 membered fused ring, a 3 to 13 membered bridged ring, and a 3 to 13 membered spiro ring.

[0294] In some aspects of Formulae E, E-l, E-2, and E-3, the linker moiety comprises a ring selected from the group consisting of Formula Fl, F2, F3, F4 and F5:

[0295] Formula F4, Formula F5,

[0296] Wherein

[0297] X' and Y' are independently selected from N and CRb;

[0298] A1, B1, C1and D1, at each occurrence, are independently selected from null, O, CO, SO, SO2, NRb, and CRbRc;

[0299] A2, B2, C2and D2, at each occurrence, are independently selected from N and CRb;

[0300] A3, B3, C3, D3, and E3at each occurrence, are independently selected from N, O, S, NRb, and CRb;

[0301] Rband Rc, at each occurrence, are independently selected from hydrogen, halogen, hydroxyl, amino, cyano, nitro, optionally substituted C1-C8alkyl, optionally substituted C2-C8alkenyl, optionally substituted C2-C8alkynyl, optionally substituted C1-C8alkoxy, optionally substituted C1-C8alkoxyalkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkylamino, optionally substituted C1-C8alkylaminoCi- C8alkyl, optionally substituted 3-10 membered carbocyclic, optionally substituted 3-10 membered cycloalkoxy, optionally substituted 3-10 membered carbocyclicamino, optionally substituted 3-8 membered heterocyclic, optionally substituted aryl, and optionally substituted heteroaryl;

[0302] Rband Rb, or Rband Rctogether with the atom to which they are connected form a 3-8 membered carbocyclic or 3-8 membered heterocyclic ring; and m1, n1, o1and p1are independently selected from 0, 1, 2, 3, 4, and 5.

[0303] In one embodiment, A, B, and W, at each occurrence, are independently selected from null, optionally substituted -(CH2)o-8-, optionally substituted -(CH2)o-8-CO-(CH2)o-8-, optionally substituted -(CH2)o-8-NH-(CH2)o-s-, optionally substituted -(CH2)o-8-NH-CO-(CH2)o-8-, optionally substituted -(CH2)o-8-CO-NH-(CH2)o-8-, optionally substituted -(CH2)o-3-NH-(CH2)o-3-CO-NH- (CH2)O-8-, optionally substituted -(CH2)O-3-NH-(CH2)I-3-NH-CO-(CH2)O-8-, optionally substituted - (CH2)O-8-CO-NH-(CH2)I-3-NH-(CH2)O-3-, optionally substituted -(CH2)o-8-Rr-(CH2)o-s-, optionally substituted -(CH2)o-3-CO-(CH2)o-3- Rr-(CH2)o-3-, -(CH2)o-8-Rr-(CH2)o-8-, optionally substituted - (CH2)O-3-NH-CO-(CH2)0-3-Rr-(CH2)0-3-, optionally substituted <CH2)o-3-NH-(CH2)0-3- Rr-(CH2)o-

[0304] In one embodiment, Rris of Formula Fl, F2, F3, F4, or F5.

[0305] In one embodiment, Rris selected from

[0306]

[0307] In another embodiment, the length of the linker is 0 to 40 atoms.

[0308] In another embodiment, the length of the linker is 0 to 20 atoms.

[0309] In another embodiment, the length of the linker is 0 to 10 atoms.

[0310] In another embodiment, the length of the linker is 0 to 40 atoms.

[0311] In another embodiment, the linker is selected from null, optionally substituted -CO-(CH2)o- io-, optionally substituted -(CH2)O-IO-, optionally substituted -(CH2)1-2-(CO)NH-(CH2)o-io-, optionally substituted -(CH2)I-2-(CO)NH-(CH2)I-3-(OCH2CH2)I-7-, optionally substituted -(CH2)o- I-CO-(CH2)1-3-(OCH2CH2)1-7-, optionally substituted -CO-(CH2)0-3-(alkenylene)-(CH2)0-3-, optionally substituted -CO-(CH2)0-3-(alkynylene)-(CH2)0-3-, optionally substituted -CO-(CH2)0-3- (3-8 membered carbocyclic)-(CH2)0-3-, optionally substituted -CO-(CH2)0-3-(3-8 membered heterocyclic)-(CH2)o-3-, optionally substituted -(CH2)o-3-(alkenylene)-(CH2)o-3-, optionally substituted -(CH2)o-3-(alkynylene)-(CH2)o-3-, optionally substituted -(CH2)o-3-(3-8 membered carbocyclic)-(CH2)o-3-, optionally substituted -(CH2)o-3-(3-8 membered heterocyclic)-(CH2)o-3-, optionally substituted -(CH2)o-s-Rr-(CH2)o-s-, optionally substituted -(CH2)o-8-Rr-(CO)-(CH2)i-8-, optionally substituted -(CH2)o-s-Rr-(CH2)i-2-(CO)-NH-(CH2)2-9-, optionally substituted -(CH2)o-8- Rr-(CH2)i-2-(CO)-NH-(CH2)i-3-(OCH2CH2)i-7-, optionally substituted -(CH2)o-s-Rr-(CH2)o-i- (CO)-(CH2)i-3-(OCH2CH2)i-7-, optionally substituted -(CH2)o-8-Rr-(CO)-(CH2)o-3-(alkenylene)- (CH2)O-3-, optionally substituted -(CH2)o-8-Rr-(CO)-(CH2)o-3-(alkynylene)-(CH2)o-3-, optionally substituted -(CH2)o-8-Rr-(CO)-(CH2)o-3-(3-8 membered carbocyclic)-(CH2)o-3-, optionally substituted -(CH2)o-8-Rr-(CO)-(CH2)o-3-(3-8 heterocyclic)-(CH2)o-3-, optionally substituted - (CH2)o-8-Rr-(CH2)o-3-(alkenylene)-(CH2)o-3-, optionally substituted -(CH2)o-8-Rr-(CH2)o-3- (alkynylene)-(CH2)o-3-, optionally substituted -(CH2)o-8-Rr-(CH2)o-3-(3-8 membered carbocyclcyl)-(CH2)o-3-, and optionally substituted -(CH2)o-8-Rr-(CH2)o-3-(heterocyclic)-(CH2)o-3--

[0312] In some embodiments, the linker can also be a moiety of:

[0313] Formula (E-2-a) Formula (E-3-a) wherein

[0314] A is C=O or CH2,

[0315] B is C=O or CH2, and m is 0-16, n is 0-6, and o is 0-6.

[0316] In some embodiments, the linker can also be a moiety of:

[0317] Formula (E-2-b), Formula (E-3-b) A is -C(O)N-, B is -C(O)N-, m is 0-16, n is 0-6, o is 0-6, n-1 is 0-6, and o-l is 0-6.

[0318] DUB Recruiter

[0319] In some aspects, the DUB recruiter comprises the structures of Formula (A-I):

[0320] Formula (A-I) or a tautomer thereof, or a pharmaceutically acceptable salt of the compound or the tautomer, wherein RD4is RD4b

[0321] In some embodiments, the DUB recruiters comprise the structures of Formulae (A-I-a), (A-I-al), (A-I-a2), (A-I-a3), (A-I-a4), (A-I-a5), (A-I-a6), (A-I-a7), (A-I-a8), (A-I-a9), (A-I-alO), (A-I-al l), (A-I-b), (A-I-bl), (A-I-b2), (A-I-b3), (A-I-b4), (A-I-b5), (A-I-b6), (A-I-b7), (A-I-b8), (A-I-b9), (A-I-blO), (A-I-bl l), (A-I-c), (A-I-cl), (A-I-c2), (A-I-c3), (A-I-c4), (A-I-c5), (A-I-c6), (A-I-C7), (A-I-C8), (A-I-C9), (A-I-c 10), (A-I-c 11), (A-I-c 12), (A-I-c 13), (A-I-c 14), (A-I-c 15), (A- I-C16), (A-I-d), (A-I-d 1), (A-I-d2), (A-I-d3), (A-I-d4), (A-I-d5), (A-I-d6), (A-I-d7), (A-I-d8), (A- I-d9), (A-I-d 10), and (A-I-b 11), wherein RD4is RD4b.

[0322] In some embodiments, RD41)is a bond.

[0323] In some embodiments, the DUB recruiters comprise:

[0324] In some embodiments, the AMPK based bivalent compound is a compound selected from the following compounds, as identified in Table 3 below: XF137-81, XF137-82, XF137-83, XF137-84, XF137-85, XF137-86, XF137-87, XF137-88, XF137-89, XF137-90, XF137-91, XF137-92, XF137-93, XF137-94, QC179-047, QC179-048, QC137-049, QC179-050, QC179- 051, QC179-052, QC179-053, QC179-054, QC137-055, and examples 367 - 380, or analogs thereof.

[0325] In some embodiments, the cGAS based bivalent compound is a compound selected from the following compounds, as identified in table 4 below: ZD178-22, ZD178-23, ZD178-24, ZD178-25, ZD178-26, ZD178-27, ZD178-28, ZD178-29, ZD178-30, ZD178-31, ZD178-32, ZD178-33, ZD178-34, ZD178-35, ZD178-58-1, ZD178-58-2, ZD178-58-3, ZD178-58-4, ZD178- 58-5, ZD178-58-6, ZD178-58-7, ZD178-58-8, ZD178-58-9, ZD178-58-10, ZD178-58-11, ZD178-58-12, ZD178-58-13, ZD178-58-14, ZD178-63-1, ZD178-63-2, ZD178-63-3, ZD178-63- 4, ZD178-63-5, ZD178-63-6, ZD178-63-78, ZD178-63-8, ZD178-63-9, ZD178-63-10, ZD178- 63-11, ZD178-63-12, ZD178-63-13, ZD178-63-14, ZD178-63-15, ZD178-69-1, ZD178-69-2, ZD178-69-3, ZD178-69-4, ZD178-69-5, ZD178-69-6, ZD178-69-7, ZD178-69-8, ZD178-69-9, ZD178-69-10, ZD178-69-11, ZD178-69-12, ZD178-69-13, ZD178-69-14, ZD178-69-15, and examples 381 - 388, or analogs thereof.

[0326] In some embodiments, the CFTR based bivalent compound is a compound selected from the following compounds, as identified in Table 5 below: QC166-130, QC166-131, QC166-132, QC166-133, QC166-134, QC166-135, QC166-136, QC166-137, QC166-138, QC166-139,

[0327] QC166-140, QC166-141, QC166-142, QC166-143, QC166-167, QC166-174, QC166-175,

[0328] QC166-176, QC166-177, QC166-178, QC166-179, QC166-181, QC166-182, QC166-183,

[0329] QC166-184, QC166-185, QC179-104, QC179-105, QC179-106, QC179-107, QC179-108,

[0330] QC179-109, QC179-110, QC179-111, QC179-112, QC179-113, QC179-137, QC179-138,

[0331] QC179-139, QC179-140, QC179-141, QC179-154, QC179-155, QC179-156, QC179-157,

[0332] QC179-158, QC192-153, QC192-154, QC192-155, QC192-156, QC192-157, QC192-184,

[0333] QC192-178, QC192-179, QC192-180, QC192-181, QC192-182, or analogs thereof.

[0334] In some embodiments, this disclosure provides a method of treating AMPK, cGAS or CFTR-mediated disease, the method including administering to a subject in need thereof one or more bivalent compounds including a AMPK, cGAS, or CFTR ligand conjugated to a 0TUB1 binder via a linker. The AMPK, cGAS, CFTR-mediated disease can be a disease resulting from AMPK, cGAS or CFTR degradation. The AMPK, cGAS or CFTR-mediated disease can have decrease CFTR expression relative to a wild-type tissue of the same species and tissue type.

[0335] In some embodiments, the methods described herein, the bivalent compounds can be administered, e.g., orally, parenterally, intradermally, subcutaneously, topically, and / or rectally.

[0336] This disclosure additionally provides a method for identifying a bivalent compound which mediates de-ubiquitination / stabilization of AMPK, cGAS, or CFTR, the method including providing a heterobifunctional test compound including a AMPK, cGAS, or CFTR ligand conjugated to a de-ubiquitination tag via a linker, contacting the heterobifunctional test compound with a cell (e.g., a cell such as a AMPK, cGAS or CFTR-mediated disease cell) including a deubiquitinase (e.g., 0TUB1) and AMPK, cGAS or CFTR protein.

[0337] As used herein, the terms “about” and “approximately” are defined as being within plus or minus 10% of a given value or state, preferably within plus or minus 5% of said value or state. The terms “bivalent” and “bi-functional” are used interchangeably herein. 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 invention; 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.

[0338] BRIEF DESCRIPTION OF THE DRAWINGS

[0339] Figure 1. Effect of exemplary compounds on modifying OTUB1 in mass spectrometry-based assays.

[0340] Figure 2. Discovery of improved OTUB1 ligand MS5105.

[0341] Figure 3. Effect of exemplary AMPK based bivalent compounds in stabilization AMPK protein levels in Hela and HEK293T cells

[0342] Figure 4. Effect of exemplary AMPK based bivalent compounds in stabilization AMPK protein level in HEK293T cells in multiple concentrations.

[0343] Figure 5. Effect of exemplary cGAS based bivalent compounds in stabilization cGAS protein level in Hela cells.

[0344] Figure 6. Discovery of first-in-class cGAS DUBTACs.

[0345] Figure 7. cGAS DUBTACs MS7829 and mS8588 activate the cGAS / STING / IRF3 signaling.

[0346] Figure 8. The MS5105-based DUBTAC MS6178 increases CFTR-AF508 protein level much more effectively than the EN523-based CFTR DUBTAC NJH-2-057.

[0347] Figure 9. Effect of exemplary CFTR based bivalent compounds on stabilizing CFTR protein levels in human cystic fibrosis bronchial epithelial cells.

[0348] Figure 10. Effect of exemplary CFTR or cGAS based bivalent compounds on modifying OTUB1 mass spectrometry-based assays.

[0349] Testing of exemplary Compounds

[0350] The activity of novel synthesized covalent OTUB1 binder and bivalent AMPK-OTUBl, cGAS-OTUBl, CFTR-OTUB1 compounds can be assessed using standard biophysical assays (e g., isothermal titration calorimetry (ITC), surface plasmon resonance (SPR)) and mass spectrometry -based assays. Cellular assays can then be used to assess the bivalent CFTR-OTUB1 compound’s ability to stabilize AMPK, cGAS or CFTR proteins level. Assays suitable for use in any or all of these steps are known in the art, and include, e.g., Western blotting, quantitative mass spectrometry (MS) analysis, flow cytometry, enzymatic inhibition, ITC, SPR, cell growth inhibition and xenograft and PDX models.

[0351] By way of non-limiting example, detailed synthesis protocols are described in the Examples for specific exemplary covalent 0TUB1 binders, bivalent AMPK-0TUB1, cGAS- 0TUB1 and CFTR-OTUB1 compounds.

[0352] Pharmaceutically acceptable isotopic variations of the compounds disclosed herein are contemplated and can be synthesized using conventional methods known in the art or methods corresponding to those described in the Examples (substituting appropriate reagents with appropriate isotopic variations of those reagents). Specifically, an isotopic variation is a compound in which at least one atom is replaced by an atom having the same atomic number, but an atomic mass different from the atomic mass usually found in nature. Useful isotopes are known in the art and include, for example, isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine. Exemplary isotopes thus include, e.g.,2H,3H,13C,14C,15N,17O,18O,32P,35S,18F, and36C1.

[0353] Isotopic variations (e.g., isotopic variations containing2H) can provide therapeutic advantages resulting from greater metabolic stability, e.g., increased in vivo half-life or reduced dosage requirements. In addition, certain isotopic variations (particularly those containing a radioactive isotope) can be used in drug or substrate tissue distribution studies. The radioactive isotopes tritium (3H) and carbon-14 (14C) are particularly useful for this purpose in view of their ease of incorporation and ready means of detection.

[0354] Pharmaceutically acceptable solvates of the compounds disclosed herein are contemplated. A solvate can be generated, e.g., by substituting a solvent used to crystallize a compound disclosed herein with an isotopic variation (e.g., D2O in place of H2O, ^-acetone in place of acetone, or de- DMSO in place of DMSO).

[0355] Pharmaceutically acceptable fluorinated variations of the compounds disclosed herein are contemplated and can be synthesized using conventional methods known in the art or methods corresponding to those described in the Examples (substituting appropriate reagents with appropriate fluorinated variations of those reagents). Specifically, a fluorinated variation is a compound in which at least one hydrogen atom is replaced by a fluoro atom. Fluorinated variations can provide therapeutic advantages resulting from greater metabolic stability, e.g., increased in vivo half-life or reduced dosage requirements. Pharmaceutically acceptable prodrugs of the compounds disclosed herein are contemplated and can be synthesized using conventional methods known in the art or methods corresponding to those described in the Examples (e.g., converting hydroxyl groups or carboxylic acid groups to ester groups). As used herein, a "prodrug" refers to a compound that can be converted via some chemical or physiological process (e.g., enzymatic processes and metabolic hydrolysis) to a therapeutic agent. Thus, the term "prodrug" also refers to a precursor of a biologically active compound that is pharmaceutically acceptable. A prodrug may be inactive when administered to a subject, i.e. an ester, but is converted hi vivo to an active compound, for example, by hydrolysis to the free carboxylic acid or free hydroxyl. The prodrug compound often offers advantages of solubility, tissue compatibility or delayed release in an organism. The term "prodrug" is also meant to include any covalently bonded carriers, which release the active compound in vivo when such prodrug is administered to a subject. Prodrugs of an active compound may be prepared by modifying functional groups present in the active compound in such a way that the modifications are cleaved, either in routine manipulation or in vivo, to the parent active compound. Prodrugs include compounds wherein a hydroxy, amino or mercapto group is bonded to any group that, when the prodrug of the active compound is administered to a subject, cleaves to form a free hydroxy, free amino or free mercapto group, respectively. Examples of prodrugs include, but are not limited to, acetate, formate and benzoate derivatives of an alcohol or acetamide, formamide and benzamide derivatives of an amine functional group in the active compound and the like.

[0356] Definition of Terms

[0357] "Alkyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation. An alkyl may comprise one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, or sixteen carbon atoms. In certain embodiments, an alkyl comprises one to fifteen carbon atoms (e.g., C1-C15 alkyl). In certain embodiments, an alkyl comprises one to thirteen carbon atoms (e.g., C1-C13 alkyl). In certain embodiments, an alkyl comprises one to eight carbon atoms (e.g., C1-C8alkyl). In other embodiments, an alkyl comprises five to fifteen carbon atoms (e.g., C5-C15 alkyl). In other embodiments, an alkyl comprises five to eight carbon atoms (e.g., Cs-Cs alkyl). The alkyl is attached to the rest of the molecule by a single bond, for example, methyl (Me), ethyl (Et), n-propyl, 1 -methylethyl ( / .w-propyl), / / -butyl, / / -pentyl, 1 , 1 -dimethylethyl ( / -butyl), pentyl, 3 -methylhexyl,

[0358] 2-methylhexyl, and the like.

[0359] “Alkylene” refers to a bivalent saturated aliphatic radical (such as ethylene) regarded as derived from an alkene by opening of the double bond or from an alkane by removal of two hydrogen atoms from different carbon atoms.

[0360] "Alkenyl1' refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one double bond. An alkenyl may comprise two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, or sixteen carbon atoms. In certain embodiments, an alkenyl comprises two to twelve carbon atoms (e.g, C2-C12alkenyl). In certain embodiments, an alkenyl comprises two to eight carbon atoms (e.g., C2-C8alkenyl). In certain embodiments, an alkenyl comprises two to six carbon atoms (e.g., C2- , alkenyl). In other embodiments, an alkenyl comprises two to four carbon atoms (e.g., C2-C4alkenyl). The alkenyl is attached to the rest of the molecule by a single bond, for example, ethenyl (i.e., vinyl), prop-l-enyl (i.e., allyl), but-l-enyl, pent-l-enyl, penta- 1,4-dienyl, and the like.

[0361] The term “alkenylene” refers to linear or branched-chain divalent hydrocarbon radical of two to eight carbon atoms (C2-C8) with at least one site of unsaturation, i.e., a carbon-carbon, SP2 double bond, wherein the alkenylene radical may be optionally substituted independently with one or more substituents described herein, and includes radicals having “cis” and “trans” orientations, or alternatively, “E” and “Z” orientations. Examples include, but are not limited to ,ethylenylene or vinylene (-CH=CH-), allyl (-CH2CH=CH-), and the like.

[0362] The term “allyl,” as used herein, means a -CH2CI-GCH2group.

[0363] As used herein, the term "alkynyl" refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one triple bond. An alkynyl may comprise two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, or sixteen carbon atoms. In certain embodiments, an alkynyl comprises two to twelve carbon atoms (e.g., C2-C12 alkynyl). In certain embodiments, an alkynyl comprises two to eight carbon atoms (e.g., C2-C8alkynyl). In other embodiments, an alkynyl has two to six carbon atoms (e.g., C2-C6alkynyl). In other embodiments, an alkynyl has two to four carbon atoms (e.g., C2-C4alkynyl). The alkynyl is attached to the rest of the molecule by a single bond. Examples of such groups include, but are not limited to, ethynyl, propynyl, 1-butynyl, 2-butynyl, 1 -pentynyl, 2-pentynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, and the like. The term “alkynylene” refers to a linear or branched divalent hydrocarbon radical of two to eight carbon atoms (C2-C8) with at least one site of unsaturation, i.e., a carbon-carbon, sp triple bond, wherein the alkynylene radical may be optionally substituted independently with one or more substituents described herein. Examples include, but are not limited to ,ethynylene (-C =C~), propynylene (propargylene, -CH2C = C-), and the like.

[0364] The term " alkoxy", as used herein, means an alkyl group as defined herein witch is attached to the rest of the molecule via an oxygen atom. Examples of such groups include, but are not limited to, methoxy, ethoxy, n-propyloxy, iso-propyloxy, n-butoxy, iso-butoxy, tert-butoxy, pentyloxy, hexyloxy, and the like.

[0365] “Heteroalkyl” refers to a substituted or unsubstituted alkyl group which has one or more skeletal chain atoms selected from an atom other than carbon, Exemplary skeletal chain atoms selected from an atom other tha n carbon include , e.g., O, N, P, Si, S, or combinations thereof, wherein the nitrogen, phosphorus, and sulfur atoms may optionally be oxidized and the nitrogen heteroatom may optionally be quaternized . If given, a numberical range refers to the chain length in total. For example, a 3- to 8- membered heteroalkyl has a chain length of 3 to 8 atoms. Connection to the rest of the molecule may be through either a heteroatom or a carbon in the heteroalkyl chain. Unless stated otherwise specifically in the specification, a heteroalkyl group is optionally substituted by one or more substituents such as those substituents described herein.

[0366] The term “aryl”, as used herein, " refers to a radical derived from an aromatic monocyclic or multicyclic hydrocarbon ring system by removing a hydrogen atom from a ring carbon atom. The aromatic monocyclic or multicyclic hydrocarbon ring system contains only hydrogen and carbon atoms. An aryl may comprise from six to eighteen carbon atoms, where at least one of the rings in the ring system is fully unsaturated, i.e., it contains a cyclic, delocalized (4n+2) 7t-electron system in accordance with the Hiickel theory. In certain embodiments, an aryl comprises six to fourteen carbon atoms (Ce-Cu aryl). In certain embodiments, an aryl comprises six to ten carbon atoms (C6-Cio aryl). Examples of such groups include, but are not limited to, phenyl, fluorenyl and naphthyl. The terms “Ph” and “phenyl,” as used herein, mean a -C6H5group.

[0367] The term “arylene” means a divalent aromatic hydrocarbon radical of 6-20 carbon atoms (C6-C20) derived by the removal of two hydrogen atom from two carbon atoms of a parent aromatic ring system. Some arylene groups are represented in the exemplary structures as “Ar”. Arylene includes bicyclic radicals comprising an aromatic ring fused to a saturated, partially unsaturated ring, or aromatic carbocyclic ring. Typical arylene groups include, but not limited to, radicals derived from benzene (phenylene), substituted benzenes, naphthalene, anthracene, biphenylene indenylene, indaylene, 1,2-dihydronaphthalene, 1,2, 3, 4, -tetrahydronaphthyl, and the like. Arylene groups are optionally substituted with one or more substituents described herein.

[0368] The term “heteroaryl”, refers to a radical derived from a 3- to 18-membered aromatic ring radical that comprises two to seventeen carbon atoms and from one to six heteroatoms selected from nitrogen, oxygen and sulfur. As used herein, the heteroaryl radical may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, wherein at least one of the rings in the ring system is fully unsaturated, i.e., it contains a cyclic, delocalized (4n+2) ^-electron system in accordance with the Hiickel theory. Heteroaryl includes fused or bridged ring systems. In certain embodiments, a heteroaryl refers to a radical derived from a 3- to 10-membered aromatic ring radical (3-10 membered heteroaryl). In certain embodiments, a heteroaryl refers to a radical derived from 5- to 7-membered aromatic ring (5-7 membered heteroaryl). Heteroaryl includes fused or bridged ring systems. The heteroatom(s) in the heteroaryl radical is optionally oxidized. One or more nitrogen atoms, if present, are optionally quatemized. The heteroaryl is attached to the rest of the molecule through any atom of the ring(s). Examples of such groups include, but not limited to, pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl,pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, furopyridinyl, and the like. In certain embodiments, an heteroaryl is attached to the rest of the molecule via a ring carbon atom. In certain embodiments, an heteroaryl is attached to the rest of the molecule via a nitrogen atom (N-attached) or a carbon atom (C-attached). For instance, a group derived from pyrrole may be pyrrol-l-yl (N-attached) or pyrrol-3-yl (C-attached). Further, a group derived from imidazole may be imidazol-l-yl (N-attached) or imidazol-3-yl (C-attached).

[0369] The term “heterocyclic”, as used herein, means a non-aromatic, monocyclic, bicyclic, tricyclic, or tetracyclic radical having a total of from 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 atoms in its ring system, and containing from 3 to 12 carbon atoms and from 1 to 4 heteroatoms each independently selected from O, S and N, and with the proviso that the ring of said group does not contain two adjacent O atoms or two adjacent S atoms. A heterocyclic group may include fused, bridged or spirocyclic ring systems. In certain embodiments, a heterocyclic group comprises 3 to 10 ring atoms (3-10 membered heterocyclic). In certain embodiments, a heterocyclic group comprises 3 to 8 ring atoms (3-8 membered heterocyclic). In certain embodiments, a heterocyclic group comprises 4 to 8 ring atoms (4-8 membered heterocyclic). In certain embodiments, a heterocyclic group comprises 3 to 6 ring atoms (3-6 membered heterocyclic). A heterocyclic group may contain an oxo substituent at any available atom that will result in a stable compound. For example, such a group may contain an oxo atom at an available carbon or nitrogen atom. Such a group may contain more than one oxo substituent if chemically feasible. In addition, it is to be understood that when such a heterocyclic group contains a sulfur atom, said sulfur atom may be oxidized with one or two oxygen atoms to afford either a sulfoxide or sulfone. An example of a 4 membered heterocyclic group is azetidinyl (derived from azetidine). An example of a 5 membered cycloheteroalkyl group is pyrrolidinyl. An example of a 6 membered cycloheteroalkyl group is piperidinyl. An example of a 9 membered cycloheteroalkyl group is indolinyl. An example of a 10 membered cycloheteroalkyl group is 4H-quinolizinyl. Further examples of such heterocyclic groups include, but are not limited to, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino, thioxanyl, piperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3- pyrrolinyl, indolinyl, 2H-pyranyl, 477- pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo[3. 1.0]hexanyl, 3-azabicyclo[4. 1.0]heptanyl, 3 / 7-indolyl, quinolizinyl, 3-oxopiperazinyl, 4-methylpiperazinyl, 4-ethylpiperazinyl, and l-oxo-2,8,diazaspiro[4.5]dec-8-yl. A heteroaryl group may be attached to the rest of molecular via a carbon atom (C-attached) or a nitrogen atom (N-attached). For instance, a group derived from piperazine may be piperazin- 1-yl (N-attached) or piperazin-2-yl (C-attached).

[0370] The term " cycloalkyl" or “carbocyclic” means a saturated, monocyclic, bicyclic, tricyclic, or tetracyclic radical having a total of from 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 carbon atoms in its ring system. A cycloalkyl may be fused, bridged or spirocyclic. In certain embodiments, a cycloalkyl comprises 3 to 8 carbon ring atoms (C3-C8cycloalkyl). In certain embodiments, a cycloalkyl comprises 3 to 6 carbon ring atoms ( C3-C6cycloalkyl). Examples of such groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cycloheptyl, adamantyl, and the like.

[0371] The term “cycloalkylene” or “carbocyclicene” is a bidentate radical obtained by removing a hydrogen atom from a cycloalkyl ring as defined above. Examples of such groups include, but are not limited to, cyclopropylene, cyclobutylene, cyclopentyl ene, cyclopentenylene, cyclohexylene, cycloheptylene, and the like.

[0372] The term "spirocyclic" as used herein has its conventional meaning, that is, any ring system containing two or more rings wherein two of the rings have one ring carbon in common. Each ring of the spirocyclic ring system, as herein defined, independently comprises 3 to 20 ring atoms. Preferably, they have 3 to 10 ring atoms. Non-limiting examples of a spirocyclic system include spiro[3.3]heptane, spiro[3.4] octane, and spiro[4.5]decane.

[0373] The term cyano" refers to a -C=N group.

[0374] An "aldehyde" group refers to a -C(O)H group.

[0375] An "alkoxy" group refers to both an -O-alkyl, as defined herein.

[0376] An "alkoxycarbonyl" refers to a -C(O)-alkoxy, as defined herein.

[0377] An "alkylaminoalkyl" group refers to an -alkyl-NR-alkyl group, as defined herein.

[0378] An "alkylsulfonyl" group refer to a -SChalkyl, as defined herein.

[0379] An "amino" group refers to an optionally substituted -NH2.

[0380] An "aminoalkyl" group refers to an -alky-amino group, as defined herein.

[0381] An "aminocarbonyl" refers to a -C(O)-amino, as defined herein.

[0382] An "arylalkyl" group refers to -alkylaryl, where alkyl and aryl are defined herein.

[0383] An "aryloxy" group refers to both an -O-aryl and an -O-heteroaryl group, as defined herein.

[0384] An "aryloxycarbonyl" refers to -C(O)-aryloxy, as defined herein.

[0385] An "aryl sulfonyl" group refers to a -SCharyl, as defined herein.

[0386] A "carbonyl" group refers to a -C(O)- group, as defined herein.

[0387] A "carboxylic acid" group refers to a -C(O)OH group.

[0388] A “cycloalkoxy” refers to a -O-cycloalkyl group, as defined herein.

[0389] A "halo" or "halogen" group refers to fluorine, chlorine, bromine or iodine.

[0390] A "haloalkyl" group refers to an alkyl group substituted with one or more halogen atoms.

[0391] A "hydroxy" group refers to an -OH group.

[0392] A "nitro" group refers to a -NO2group. An “oxo” group refers to the =0 substituent.

[0393] A "trihalomethyl" group refers to a methyl substituted with three halogen atoms.

[0394] The term “substituted,” means that the specified group or moiety bears one or more substituents independently selected from C1-C4alkyl, aryl, heteroaryl, aryl-C1-C4alkyl-, heteroaryl-C1-C4alkyl-, C1-C4haloalkyl, -OC1-C4alkyl, -OC1-C4alkylphenyl, -C1-C4alkyl-OH, -OC1-C4haloalkyl, halo, -OH, -NH2, -C1-C4alkyl-NH2, -N(C1-C4alkyl)(C1-C4alkyl), -NH(C1-C4alkyl), -N(C1-C4alkyl)(C1-C4alkylphenyl), -NH(C1-C4alkylphenyl), cyano, nitro, oxo, -C02H, -C(O)OC1-C4alkyl, -CON(C1-C4alkyl)(C1-C4alkyl), -CONH(C1-C4alkyl), -C0NH2, -NHC(O)(C1-C4alkyl), -NHC(O)(phenyl), -N(C1-C4alkyl)C(O)(C1-C4alkyl), -N(C1-C4alkyl)C(O)(phenyl), -C(O)C1-C4alkyl, -C(O)C1-C4alkylphenyl, -C(O)C1-C4haloalkyl, -OC(O)C1-C4alkyl, -SO2(C1-C4alkyl), -SO2(phenyl), -SO2(C1-C4haloalkyl), -SO2NH2, -SO2NH(C1-C4alkyl), -SO2NH(phenyl), -NHSO2(C1-C4alkyl), -NHSO2(phenyl), and -NHSO2(C1-C4haloalkyl).

[0395] The term “null” means the absence of an atom or moiety, and there is a bond between adjacent atoms in the structure.

[0396] The term “optionally substituted” means that the specified group may be either unsubstituted or substituted by one or more substituents as defined herein. It is to be understood that in the compounds of the present invention when a group is said to be “unsubstituted,” or is “substituted” with fewer groups than would fill the valencies of all the atoms in the compound, the remaining valencies on such a group are filled by hydrogen. For example, if a Ce aryl group, also called “phenyl” herein, is substituted with one additional substituent, one of ordinary skill in the art would understand that such a group has 4 open positions left on carbon atoms of the C6aryl ring (6 initial positions, minus one at which the remainder of the compound of the present invention is attached to and an additional substituent, remaining 4 positions open). In such cases, the remaining 4 carbon atoms are each bound to one hydrogen atom to fill their valencies. Similarly, if a Ce aryl group in the present compounds is said to be “di substituted,” one of ordinary skill in the art would understand it to mean that the C6aryl has 3 carbon atoms remaining that are unsubstituted. Those three unsubstituted carbon atoms are each bound to one hydrogen atom to fill their valencies. As used herein, the same symbol in a different FORMULA may have a different definition, for example, the definition of R1 in FORMULA 1 is as defined with respect to FORMULA 1 and the definition of R1 in FORMULA 6 is as defined with respect to FORMULA 6.

[0397] As used herein, when m (or n or u or v or w) is defined by a range, for example, “m is 0 to 15” or “m = 0-3” means that m is an integer from 0 to 15 (i.e. m is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15) or m is an integer from 0 to 3(i.e. m is 0, 1,2, or 3) or is any integer in the defined range.

[0398] "Pharmaceutically acceptable salt" includes both acid and base addition salts. A pharmaceutically acceptable salt of any one of the bivalent compounds described herein is intended to encompass any and all pharmaceutically suitable salt forms. Preferred pharmaceutically acceptable salts of the compounds described herein are pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

[0399] "Pharmaceutically acceptable acid addition salt" refers to those salts which retain the biological effectiveness and properties of the free bases, which are not biologically or otherwise undesirable, and which are formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, hydroiodic acid, hydrofluoric acid, phosphorous acid, and the like. Also included are salts that are formed with organic acids such as aliphatic mono- and dicarboxylic acids, phenyl -substituted alkanoic acids, hydroxy alkanoic acids, alkanedioic acids, aromatic acids, aliphatic and. aromatic sulfonic acids, etc. and include, for example, acetic acid, tri fluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. Exemplary salts thus include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates, monohydrogenphosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, trifluoroacetates, propionates, caprylates, isobutyrates, oxalates, malonates, succinate suberates, sebacates, fumarates, maleates, mandelates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, phthalates, benzenesulfonates, toluenesulfonates, phenylacetates, citrates, lactates, malates, tartrates, methanesulfonates, and the like. Also contemplated are salts of amino acids, such as arginates, gluconates, and galacturonates (see, for example, Berge S.M. et al., "Pharmaceutical Salts," Journal of Pharmaceutical Science, 66: 1-19 (1997), which is hereby incorporated by reference in its entirety). Acid addition salts of basic compounds may be prepared by contacting the free base forms with a sufficient amount of the desired acid to produce the salt according to methods and techniques with which a skilled artisan is familiar.

[0400] "Pharmaceutically acceptable base addition salt" refers to those salts that retain the biological effectiveness and properties of the free acids, which are not biologically or otherwise undesirable. These salts are prepared from addition of an inorganic base or an organic base to the free acid. Pharmaceutically acceptable base addition salts may be formed with metals or amines, such as alkali and alkaline earth metals or organic amines. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts and the like. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, for example, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, N '.N-dibenzylethylenediamine, chloroprocaine, hydrabamine, choline, betaine, ethylenediamine, ethylenedianiline, A-methylglucamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine, A-ethylpiperidine, polyamine resins and the like. See Berge et al., supra.

[0401] Pharmaceutical Compositions

[0402] In some aspects, the compositions and methods described herein include the manufacture and use of pharmaceutical compositions and medicaments that include one or more bivalent compounds as disclosed herein. Also included are the pharmaceutical compositions themselves.

[0403] In some aspects, the compositions disclosed herein can include other compounds, drugs, or agents used for the treatment of cancer. For example, in some instances, pharmaceutical compositions disclosed herein can be combined with one or more (e.g., one, two, three, four, five, or less than ten) compounds. Such additional compounds can include, e.g., conventional chemotherapeutic agents known in the art. When co-administered, 0TUB1 covalent binders or AMPK, cGAS or CFTR based bivalent compounds disclosed herein can operate in conjunction with conventional chemotherapeutic agents to produce mechanistically additive or synergistic therapeutic effects. In some aspects, the pH of the compositions disclosed herein can be adjusted with pharmaceutically acceptable acids, bases, or buffers to enhance the stability of the 0TUB1 covalent binders or AMPK, cGAS or CFTR based bivalent compounds or its delivery form.

[0404] Pharmaceutical compositions typically include a pharmaceutically acceptable carrier, adjuvant, or vehicle. As used herein, the phrase “pharmaceutically acceptable” refers to molecular entities and compositions that are generally believed to be physiologically tolerable and do not typically produce an allergic or similar untoward reaction, such as gastric upset, dizziness and the like, when administered to a human. A pharmaceutically acceptable carrier, adjuvant, or vehicle is a composition that can be administered to a patient, together with a compound of the invention, and which does not destroy the pharmacological activity thereof and is nontoxic when administered in doses sufficient to deliver a therapeutic amount of the compound. Exemplary conventional nontoxic pharmaceutically acceptable carriers, adjuvants, and vehicles include saline, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration.

[0405] In particular, pharmaceutically acceptable carriers, adjuvants, and vehicles that can be used in the pharmaceutical compositions of this invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, self-emulsifying drug delivery systems (SEDDS) such as d- a-tocopherol polyethylene glycol 1000 succinate, surfactants used in pharmaceutical dosage forms such as Tweens or other similar polymeric delivery matrices, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene polyoxypropyl ene-block polymers, polyethylene glycol and wool fat. Cyclodextrins such as a-, -, and γ-cyclodextrin, may also be advantageously used to enhance delivery of compounds of the formulae described herein.

[0406] As used herein, the OTUB 1 covalent binders or AMPK, cGAS or CFTR based bivalent compounds disclosed herein are defined to include pharmaceutically acceptable derivatives or prodrugs thereof. A “pharmaceutically acceptable derivative” means any pharmaceutically acceptable salt, solvate, or prodrug, e.g., carbamate, ester, phosphate ester, salt of an ester, or other derivative of a compound or agent disclosed herein, which upon administration to a recipient is capable of providing (directly or indirectly) a compound described herein, or an active metabolite or residue thereof. Particularly favored derivatives and prodrugs are those that increase the bioavailability of the compounds disclosed herein when such compounds are administered to a mammal (e.g., by allowing an orally administered compound to be more readily absorbed into the blood) or which enhance delivery of the parent compound to a biological compartment (e.g., the brain or lymphatic system) relative to the parent species. Preferred prodrugs include derivatives where a group that enhances aqueous solubility or active transport through the gut membrane is appended to the structure of formulae described herein. Such derivatives are recognizable to those skilled in the art without undue experimentation. Nevertheless, reference is made to the teaching of Burger’s Medicinal Chemistry and Drug Discovery, 5thEdition, Vol. 1: Principles and Practice, which is incorporated herein by reference to the extent of teaching such derivatives.

[0407] The OTUB1 covalent binders or AMPK, cGAS or CFTR based bivalent compounds disclosed herein include pure enantiomers, mixtures of enantiomers, pure diastereoisomers, mixtures of diastereoisomers, diastereoisomeric racemates, mixtures of diastereoisomeric racemates and the meso-form and pharmaceutically acceptable salts, solvent complexes, morphological forms, or deuterated derivatives thereof.

[0408] In particular, pharmaceutically acceptable salts of the OTUB1 covalent binders or AMPK, cGAS or CFTR based bivalent compounds disclosed herein include, e.g., those derived from pharmaceutically acceptable inorganic and organic acids and bases. Examples of suitable acid salts include acetate, adipate, benzoate, benzenesulfonate, butyrate, citrate, digluconate, dodecyl sulfate, formate, fumarate, glycolate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, lactate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, palmoate, phosphate, picrate, pivalate, propionate, salicylate, succinate, sulfate, tartrate, tosylate, trifluoromethylsulfonate, and undecanoate. Salts derived from appropriate bases include, e.g., alkali metal (e.g., sodium), alkaline earth metal (e.g., magnesium), ammonium and N- (alkyl)4+ salts. The invention also envisions the quaternization of any basic nitrogen-containing groups of the OTUB1 covalent binders or AMPK, cGAS and CFTR based bivalent compounds disclosed herein. Water or oil-soluble or dispersible products can be obtained by such quaternization. In some aspects, the pharmaceutical compositions disclosed herein can include an effective amount of one or more OTUB1 covalent binders or AMPK, cGAS and CFTR based bivalent compounds. The terms “effective amount” and “effective to treat,” as used herein, refer to an amount or a concentration of one or more compounds or a pharmaceutical composition described herein utilized for a period of time (including acute or chronic administration and periodic or continuous administration) that is effective within the context of its administration for causing an intended effect or physiological outcome (e.g., treatment or prevention of cell growth, cell proliferation, or cancer). In some aspects, pharmaceutical compositions can further include one or more additional compounds, drugs, or agents used for the treatment of cancer (e.g., conventional chemotherapeutic agents) in amounts effective for causing an intended effect or physiological outcome (e g., treatment or prevention of cell growth, cell proliferation, or cancer).

[0409] In some aspects, the pharmaceutical compositions disclosed herein can be formulated for sale in the United States, import into the United States, or export from the United States.

[0410] Administration of Pharmaceutical Compositions

[0411] The pharmaceutical compositions disclosed herein can be formulated or adapted for administration to a subject via any route, e.g., any route approved by the Food and Drug Administration (FDA). Exemplary methods are described in the FDA Data Standards Manual (DSM) (available at http: / / www.fda.gov / Drugs / DevelopmentApprovalProcess / FormsSubmissionRequirements / ElectronicSubmissions / DataStandardsManualmonographs). In particular, the pharmaceutical compositions can be formulated for and administered via oral, parenteral, or transdermal delivery. The term “parenteral” as used herein includes subcutaneous, intracutaneous, intravenous, intramuscular, intraperitoneal, intra-articular, intra-arterial, intrasynovial, intrastemal, intrathecal, intralesional, and intracranial injection or infusion techniques.

[0412] For example, the pharmaceutical compositions disclosed herein can be administered, e.g., topically, rectally, nasally (e.g., by inhalation spray or nebulizer), buccally, vaginally, subdermally (e.g., by injection or via an implanted reservoir), or ophthalmically.

[0413] For example, pharmaceutical compositions of this invention can be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, emulsions and aqueous suspensions, dispersions and solutions. In the case of tablets for oral use, carriers which are commonly used include lactose and com starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried com starch. When aqueous suspensions or emulsions are administered orally, the active ingredient may be suspended or dissolved in an oily phase is combined with emulsifying or suspending agents. If desired, certain sweetening, flavoring, or coloring agents can be added.

[0414] For example, the pharmaceutical compositions of this invention can be administered in the form of suppositories for rectal administration. These compositions can be prepared by mixing a compound of this invention with a suitable non-irritating excipient which is solid at room temperature but liquid at the rectal temperature and therefore will melt in the rectum to release the active components. Such materials include, but are not limited to, cocoa butter, beeswax, and polyethylene glycols.

[0415] For example, the pharmaceutical compositions of this invention can be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and can be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, or other solubilizing or dispersing agents known in the art.

[0416] For example, the pharmaceutical compositions of this invention can be administered by injection (e.g., as a solution or powder). Such compositions can be formulated according to techniques known in the art using suitable dispersing or wetting agents (such as, for example, Tween 80) and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, e.g., as a solution in 1,3 -butanediol. Among the acceptable vehicles and solvents that may be employed are mannitol, water, Ringer’s solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil can be employed, including synthetic mono- or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically- acceptable oils, e.g., olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions can also contain a long-chain alcohol diluent or dispersant, or carboxymethyl cellulose or similar dispersing agents which are commonly used in the formulation of pharmaceutically acceptable dosage forms such as emulsions and or suspensions. Other commonly used surfactants such as Tweens, Spans, or other similar emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms can also be used for the purposes of formulation.

[0417] In some aspects, an effective dose of a pharmaceutical composition of this invention can include, but is not limited to, e.g., about 0.00001, 0.0001, 0.001, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1, 1.25, 1.5, 1.75, 2, 2.5, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 2500, 5000, or 10000 mg / kg / day, or according to the requirements of the particular pharmaceutical composition.

[0418] When the pharmaceutical compositions disclosed herein include a combination of a compound of the formulae described herein (e.g., an OTUB1 covalent binder or an AMPK, cGAS or CFTR based bivalent compound) and one or more additional compounds (e.g., one or more additional compounds, drugs, or agents used for the treatment of cancer or any other condition or disease, including conditions or diseases known to be associated with or caused by cancer), both the compound and the additional compound should be present at dosage levels of between about 1 to 100%, and more preferably between about 5 to 95% of the dosage normally administered in a monotherapy regimen. The additional agents can be administered separately, as part of a multiple dose regimen, from the compounds of this invention. Alternatively, those agents can be part of a single dosage form, mixed together with the compounds of this invention in a single composition.

[0419] In some aspects, the pharmaceutical compositions disclosed herein can be included in a container, pack, or dispenser together with instructions for administration.

[0420] Methods of Treatment

[0421] The methods disclosed herein contemplate administration of an effective amount of a compound or composition to achieve the desired or stated effect. Typically, the compounds or compositions of the invention will be administered from about 1 to about 6 times per day or, alternately or in addition, as a continuous infusion. Such administration can be used as a chronic or acute therapy. The amount of active ingredient that can be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. A typical preparation will contain from about 5% to about 95% active compound (w / w). Alternatively, such preparations can contain from about 20% to about 80% active compound. In some aspects, the present disclosure provides methods for using a composition comprising an 0TUB1 covalent binder or AMPK, cGAS or CFTR based bivalent compound, including pharmaceutical compositions (indicated below as ‘X’) disclosed herein in the following methods:

[0422] Substance X for use as a medicament in the treatment of one or more diseases or conditions disclosed herein (e.g., cancer, referred to in the following examples as ‘Y’). Use of substance X for the manufacture of a medicament for the treatment of Y; and substance X for use in the treatment of Y.

[0423] In some aspects, the methods disclosed include the administration of a therapeutically effective amount of one or more of the compounds or compositions described herein to a subject (e.g., a mammalian subject, e.g., a human subject) who is in need of, or who has been determined to be in need of, such treatment. In some aspects, the methods disclosed include selecting a subject and administering to the subject an effective amount of one or more of the compounds or compositions described herein, and optionally repeating administration as required for the prevention or treatment of cancer.

[0424] In some aspects, subject selection can include obtaining a sample from a subject (e.g., a candidate subject) and testing the sample for an indication that the subject is suitable for selection. In some aspects, the subject can be confirmed or identified, e.g. by a health care professional, as having had or having a condition or disease. In some aspects, suitable subjects include, for example, subjects who have or had a condition or disease but that resolved the disease or an aspect thereof, present reduced symptoms of disease (e.g., relative to other subjects (e.g., the majority of subjects) with the same condition or disease), or that survive for extended periods of time with the condition or disease (e.g., relative to other subjects (e.g., the majority of subjects) with the same condition or disease), e.g., in an asymptomatic state (e.g., relative to other subjects (e.g., the majority of subjects) with the same condition or disease). In some aspects, exhibition of a positive immune response towards a condition or disease can be made from patient records, family history, or detecting an indication of a positive immune response. In some aspects, multiple parties can be included in subject selection. For example, a first party can obtain a sample from a candidate subject and a second party can test the sample. In some aspects, subjects can be selected or referred by a medical practitioner (e.g., a general practitioner). In some aspects, subject selection can include obtaining a sample from a selected subject and storing the sample or using the in the methods disclosed herein. Samples can include, e.g., cells or populations of cells.

[0425] In some aspects, methods of treatment can include a single administration, multiple administrations, and repeating administration of one or more compounds disclosed herein as required for the prevention or treatment of the disease or condition from which the subject is suffering (e.g., AMPK, cGAS or CFTR-related disease). In some aspects, methods of treatment can include assessing a level of disease in the subject prior to treatment, during treatment, or after treatment. In some aspects, treatment can continue until a decrease in the level of disease in the subject is detected.

[0426] The term “subject,” as used herein, refers to any animal. In some instances, the subject is a mammal. In some instances, the term “subject,” as used herein, refers to a human (e.g., a man, a woman, or a child).

[0427] The terms “administer,” “administering,” or “administration,” as used herein, refer to implanting, ingesting, injecting, inhaling, or otherwise absorbing a compound or composition, regardless of form. For example, the methods disclosed herein include administration of an effective amount of a compound or composition to achieve the desired or stated effect.

[0428] The terms “treat”, “treating,” or “treatment,” as used herein, refer to partially or completely alleviating, inhibiting, ameliorating, or relieving the disease or condition from which the subject is suffering. This means any manner in which one or more of the symptoms of a disease or disorder (e.g., cancer) are ameliorated or otherwise beneficially altered. As used herein, amelioration of the symptoms of a particular disorder (e.g., cancer) refers to any lessening, whether permanent or temporary, lasting or transient that can be attributed to or associated with treatment by the compositions and methods of the present invention. In some embodiments, treatment can promote or result in, for example, a decrease in the number of tumor cells (e.g., in a subject) relative to the number of tumor cells prior to treatment; a decrease in the viability (e.g., the average / mean viability) of tumor cells (e.g., in a subject) relative to the viability of tumor cells prior to treatment; a decrease in the rate of growth of tumor cells; a decrease in the rate of local or distant tumor metastasis; or reductions in one or more symptoms associated with one or more tumors in a subject relative to the subject’s symptoms prior to treatment.

[0429] The terms “prevent,” “preventing,” and “prevention,” as used herein, shall refer to a decrease in the occurrence of a disease or decrease in the risk of acquiring a disease or its associated symptoms in a subject. The prevention may be complete, e.g., the total absence of disease or pathological cells in a subject. The prevention may also be partial, such that the occurrence of the disease or pathological cells in a subject is less than, occurs later than, or develops more slowly than that which would have occurred without the present invention. Exemplary AMPK, cGAS or CFTR-mediated diseases that can be treated with AMPK, cGAS or CFTR based DUBTACs include, for example, cystic fibrosis, breast cancer, ovarian cancer, prostate cancer, colon cancer, pancreatic cancer, bladder cancer, liver cancer and cervical cancer.

[0430] As used herein, the term “preventing a disease” (e g., preventing cancer) in a subject means for example, to stop the development of one or more symptoms of a disease in a subject before they occur or are detectable, e.g., by the patient or the patient’s doctor. Preferably, the disease (e.g., cancer) does not develop at all, i.e., no symptoms of the disease are detectable. However, it can also result in delaying or slowing of the development of one or more symptoms of the disease. Alternatively, or in addition, it can result in the decreasing of the severity of one or more subsequently developed symptoms.

[0431] Specific dosage and treatment regimens for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health status, sex, diet, time of administration, rate of excretion, drug combination, the severity and course of the disease, condition or symptoms, the patient’s disposition to the disease, condition or symptoms, and the judgment of the treating physician.

[0432] An effective amount can be administered in one or more administrations, applications or dosages. A therapeutically effective amount of a therapeutic compound (i.e., an effective dosage) depends on the therapeutic compounds selected. Moreover, treatment of a subject with a therapeutically effective amount of the compounds or compositions described herein can include a single treatment or a series of treatments. For example, effective amounts can be administered at least once. The compositions can be administered from one or more times per day to one or more times per week; including once every other day. The skilled artisan will appreciate that certain factors can influence the dosage and timing required to effectively treat a subject, including but not limited to the severity of the disease or disorder, previous treatments, the general health or age of the subject, and other diseases present.

[0433] Following administration, the subject can be evaluated to detect, assess, or determine their level of disease. In some instances, treatment can continue until a change (e.g., reduction) in the level of disease in the subject is detected. Upon improvement of a patient’s condition (e.g., a change (e.g., decrease) in the level of disease in the subject), a maintenance dose of a compound, or composition disclosed herein can be administered, if necessary. Subsequently, the dosage or frequency of administration, or both, can be reduced, e.g., as a function of the symptoms, to a level at which the improved condition is retained. Patients may, however, require intermittent treatment on a long-term basis upon any recurrence of disease symptoms.

[0434] The present disclosure is also described and demonstrated by way of the following examples. However, the use of these and other examples anywhere in the specification is illustrative only and in no way limits the scope and meaning of the invention or of any exemplified term. Likewise, the invention is not limited to any particular preferred embodiment or aspect described herein. Indeed, many modifications and variations may be apparent to those skilled in the art upon reading this specification, and such variations can be made without departing from the invention in spirit or in scope. The invention is therefore to be limited only by the terms of the appended claims along with the full scope of equivalents to which those claims are entitled.

[0435] EXAMPLES

[0436] Procedures for the synthesis of OTUB1 covalent binders

[0437] Scheme 1. Synthesis of example 1

[0438] Example 1

[0439] Example 1: 4-acryloyl-l-(5-methylthiophen-2-yl)piperazin-2-one (XS154-91) To a solution of 2-bromo-5-methylthiophene (50.0 mg, 0.28 mmol) dissolved in dioxane (1 mL), N,N’- dimethylethylenediamine ( 9 pL, 0.0846 mmol, 0.3 eq), K2CO3( 116.9 mg, 0.846 mmol, 3.0 eq), Cui ( 5.4 mg, 0.028 mmol, O.leq ) followed by tert-butyl 3 -oxopiperazine- 1 -carboxylate (84.7 mg, 0.42 mmol, 1.5 eq). The reaction mixture was stirred at 100 °C under nitrogen atmosphere overnight. After cooling down to rt, resulting crude mixtures were purified via silica gel column chromatography to yield intermediate 1. To a solution of intermediate 1 (20.4 mg, 0.069 mmol) dissolved in DCM (0.5 mL), and the TFA( 0.5 mL) were added. The reaction mixture stirred at rt 1 h. Excess TFA was removed, resulting crude product was re-dissovled in DCM (1 mL), Et3N (13 pL, 0.089 mmol, 2.0 eq) was added. At 0 °C the acryloyl chloride (4.8 pL, 0.053 mmol, 1.2 eq.) was added, the reaction mixture stirred at 0 °C for 30 mins. Resulting crude mixtures were purified via prep-HPLC to yield title compound (White solid, 55% yield) 'HNMR (400 MHz, Methanol-d4) 8 6.92 - 6.68 (m, 1H), 6.68 - 6.54 (m, 2H), 6.31 (d, J = 16.7 Hz, 1H), 5.90 - 5.76 (m, 1H), 4.48 (d, J = 28.0 Hz, 2H), 4.15 - 3.81 (m, 4H), 2.42 (d, J= 2.2 Hz, 3H). MS (ESI) [M+H]+=251.0.

[0440] Example 2: 4-acryloyl-l-(p-tolyl)piperazin-2-one (XS154-130). Example 2 was synthesized following similar procedure for preparing example 1. White solid, 32% yield.TH NMR (400 MHz,

[0441] Methanol-^) 6 7.31 - 7.20 (m, 4H), 6.92 - 6.69 (m, 1H), 6.33 (dt, J= 16.7, 2.4 Hz, 1H), 5.85 (d,

[0442] J= 10.6 Hz, 1H), 4.54 - 4.30 (m, 2H), 4.12 - 3.97 (m, 2H), 3.88 - 3.74 (m, 2H), 2.37 (d, J= 2.2

[0443] Example 4: 4-acryloyl-l-(2-methylpyrimidin-4-yl)piperazin-2-one (XS154-149). Example 4 was synthesized following similar procedure for preparing example 1. White solid, 46% yield.1H NMR (400 MHz, Chloroform-d / ) 8 8.68 (d, J= 6.3 Hz, 1H), 8.35 (s, 1H), 6.54 (t, J= 13.2 Hz, 1H), 6.44 (d, J = 16.4 Hz, 1H), 5.85 (dd, J = 10.0, 2.6 Hz, 1H), 4.64 - 4.45 (m, 2H), 4.44 - 4.26 (m, 2H), 4.05 - 3.87 (m, 2H), 2.77 (d, J= 2.2 Hz, 3H). MS (ESI) [M+H]+=247.3.

[0444] Scheme 2. Synthesis of example 5

[0445] Example 5: l-(4-(» / -tolyl)piperazin-l-yl)prop-2-en-l-one (XS159-19). To a solution of the 1- iodo-3 -methylbenzene (218.0 mg, 1 mmol) dissolved in dioxane (3 mL), Xantphos (173 mg, 0.3 mmol, 0.3 eq), Cs2CO3(655.6 mg, 2.0 eq), Pd2(dba)3(91.6 mg, 0.1 mmol, 0.1 eq), followed by / cvZ-butyl 3 -oxopiperazine- 1 -carboxylate (279.6 mg, 1.5 mmol, 1.5 eq). The reaction mixture was stirred at 100 °C under nitrogen atmosphere overnight. After cooling down to rt, resulting crude mixtures were purified via silica gel column chromatography to yield intermediate 2.

[0446] To a solution of intermediate 2 (260 mg, 0.94 mmol) dissolved in DCM (1 mL), and the TFA( 1 mL) were added. The reaction mixture stirred at rt 1 h. Excess TFA was removed, resulting crude product was re-dissovled in DCM (1 mL), Et3N (380 pL, 2.68 mmol, 3.0 eq) was added. At 0 °C the acryloyl chloride (87 pL, 1.07 mmol, 1.2 eq.) was added, the reaction mixture stirred at 0 °C for 30 mins. Resulting crude mixtures were purified via prep-HPLC to yield title compound (White solid, 17% yield) *HNMR (400 MHz, DMSO-d6) 87.21 - 7.07 (m, 1H), 6.91 - 6.76 (m, 3H), 6.71 (d, J = 7.6 Hz, 1H), 6.14 (dd, J = 16.6, 3.1 Hz, 1H), 5.71 (dd, J = 10.4, 3.0 Hz, 1H), 3.78 - 3.65 (m, 4H), 3.17 (s, 4H), 2.26 (d, J= 2.3 Hz, 3H). MS (ESI) [M+H]+=231.1

[0447] Scheme 3. Synthesis of example 6

[0448] Example 6: 3-(3-(4-acryloyl-2-oxopiperazin-l-yl)phenyl)propanoic acid (XS159-13). To a solution of tert-butyl 3-(3-iodophenyl)propanoate (290.1 mg, 1 mmol) dissolved in dioxane (3 mL), N,N’ -dimethylethylenediamine ( 32 pL, 0.3 mmol, 0.3 eq), K2CO3( 414.6 mg, 3.0 mmol, 3.0 eq), Cui ( 19.1 mg, 0.1 mmol, 0. leq ) followed by tert-butyl 3 -oxopiperazine- 1 -carboxylate (300.3 mg, 1.5 mmol, 1.5 eq). The reaction mixture was stirred at 100 °C under nitrogen atmosphere overnight. After cooling down to rt, resulting crude mixtures were purified via silica gel column chromatography to yield intermediate 3.

[0449] To a solution of intermediate 3 (130 mg, 0.36 mmol) dissolved in DCM (0.5 mL), and the TFA( 0.5 mL) were added. The reaction mixture stirred at rt 1 h. Excess TFA was removed, resulting crude product was re-dissovled in DCM (1 mL), Et3N(68 pL, 0.483 mmol, 3.0 eq) was added. At 0 °C the acryloyl chloride (16 pL, 0.194 mmol, 1.2 eq.) was added, the reaction mixture stirred at 0 °C for 30 mins. Resulting crude mixtures were purified via prep-HPLC to yield title compound (White solid, 37% yield) 'H NMR (400 MHz, Methanol-d 4 7.35 (t, J= 6.3 Hz, 1H), 7.24 - 7.16 (m, 3H), 6.90 - 6.66 (m, 1H), 6.32 (dd, J= 16.9, 3.7 Hz, 1H), 5.84 (d, J= 10.5 Hz, 1H), 4.43 (d, J = 25.3 Hz, 2H), 4.04 (t, J= 5.9 Hz, 2H), 3.90 - 3.77 (m, 2H), 2.95 (q, J= 1A, 6.5 Hz, 2H), 2.63 (q, J= 7.5, 5.6 Hz, 2H). MS (ESI) [M+H]+=303.0.

[0450] Example 7: l-acryloyl-4-(5-methylfuran-2-yl)-l,4-diazepan-5-one (XS159-107). Example 7 was synthesized following similar procedure for preparing example 1. White solid, 27% yield. 'H NMR (400 MHz, Methanol-^) 8 7.43 - 6.67 (m, 2H), 6.23 (d, J= 17.0 Hz, 1H), 6.03 (d, J= 13.7 Hz, 1H), 5.79 (s, 1H), 3.92 - 3.76 (m, 6H), 2.73 - 2.63 (m, 2H), 2.25 (s, 3H). MS (ESI) [M+H]+=249.2.

[0451] Example 8: 4-:icryloyl-l-( L2-diinethyl-l / / -iniidazol-5-yl)piperazin-2-one (XS165-30).

[0452] Example 8 was synthesized following similar procedure for preparing example 1. White solid, 19% yield.JH NMR (400 MHz, Chloroform-d / ) δ 7.38 (d, J= 20.7 Hz, 1H), 6.65 - 6.41 (m, 1H), 6.34 (d, J= 16.6 Hz, 1H), 5.79 (d, J= 10.4 Hz, 1H), 4.41 (s, 2H), 3.98 (d, J= 19.4 Hz, 4H), 3.69 (s, 3H), 2.58 (s, 3H). MS (ESI) [M+H]+=235.3.

[0453] Example 9: 4-acryloyl-l-(6-methylpyridin-2-yl)piperazin-2-one (XS159-90) . Example 9 was synthesized following similar procedure for preparing example 1. White solid, 53% yield. 'H NMR (400 MHz, Chloroform -d) 8 7.82 - 7.65 (m, 1H), 7.60 (t, J= 7.8 Hz, 1H), 6.98 (d, J= 7.5 Hz, 1H), 6.63 - 6.46 (m, 1H), 6.40 (dd, J = 16.7, 1.9 Hz, 1H), 5.89 - 5.69 (m, 1H), 4.44 (d, J = 27.3 Hz, 2H), 4.20 (d, J= 29.6 Hz, 2H), 3.94 (d, J= 25.4 Hz, 2H), 2.49 (s, 3H). MS (ESI) [M+H]+=246.4. Example 12: 1 -(3-(zM-tolyl)-3,8-diazabicyclo[3.2.1 ] octan-8-yl)prop-2-en-1 -one(XS 165-33). Example 12 was synthesized following similar procedure for preparing example 5. White solid, 13% yield. ’H NMR (400 MHz, DMSO-t76) 8 7.05 (t, J= 7.8 Hz, 1H), 6.80 - 6.70 (m, 1H), 6.69 - 6.60 (m, 2H), 6.56 (d, J= 7.5 Hz, 1H), 6. 17 (dt, J= 16.5, 2.3 Hz, 1H), 5.69 (dt, J= 10.2, 2.3 Hz, 1H), 4.73 - 4.50 (m, 2H), 3.52 (t, J= 13.1 Hz, 2H), 2.71 (dd, J= 25.4, 11.2 Hz, 2H), 2.54 - 2.45 (m, 4H), 2.22 (s, 3H), 1.88 (dd, J= 17.7, 7.5 Hz, 2H), 1.79 (d, J= 8.3 Hz, 2H). MS (ESI) [M+H]- =257.5.

[0454] Example 13: l-(8-(m-tolyl)-3,8-diazabicydo[3.2.1 ]octan-3-yl)prop-2-en-l-one (XS165-54). Example 13 was synthesized following similar procedure for preparing example 5. White solid, 17% yield. *H NMR (400 MHz, Methanol-^) 8 7.15 - 7.06 (m, 1H), 6.82 (s, 1H), 6.77 (d, J= 8.3 Hz, 1H), 6.69 - 6.55 (m, 2H), 6.15 (dt, J= 16.8, 2.0 Hz, 1H), 5.68 (dd, J= 10.6, 2.0 Hz, 1H), 4.31 (d, J= 5.3 Hz, 2H), 4.21 (d, J= 13.5 Hz, 1H), 3.75 (d, J= 13.0 Hz, 1H), 3.53 (d, J= 13.1 Hz, 1H), 3.10 (d, J = 13.4 Hz, 1H), 2.25 (s, 3H), 1.96 (t, J= 6.3 Hz, 2H), 1.77 - 1.63 (m, 2H). MS (ESI)

[0455] Example 14: 4-acryloyl-l-(2-methylthiazol-5-yl)piperazin-2-one (XS154-184). Example 13 was synthesized following similar procedure for preparing example 1. White solid, 43% yield. 'H NMR (400 MHz, Methanol-d4) 8 7.66 (d, J= 44.8 Hz, 1H), 6.94 - 6.68 (m, 1H), 6.32 (d, J= 16.6

[0456] Hz, 1H), 5.85 (d, .7= 9.2 Hz, 1H), 4.74 - 4.48 (m, 3H), 4.16 - 3.93 (m, 4H), 2.67 (s, 3H). MS (ESI) [M+H]+=252.0. Example 15: 4-(but-2-ynoyl)-l-(m-tolyl)piperazin-2-one (XS165-75). Example 15 was synthesized following similar procedure for preparing example 1. Colorless oil, 19% yield. ’H NMR (400 MHz, Methanol-^) 6 7.36 - 7.25 (m, 1H), 7.20 - 7.03 (m, 3H), 4.54 (d, J= 1.9 Hz, 1H), 4.32 (d, J= 1.8 Hz, 1H), 4.16 (td, J= 5.6, 1.9 Hz, 1H), 4.00 - 3.90 (m, 1H), 3.83 - 3.79 (m, 1H), 3.78 - 3.72 (m, 1H), 2.36 (d, J= 1.8 Hz, 3H), 2.07 (dd, J= 8.9, 1.8 Hz, 3H). MS (ESI) [M+H]+=252.5.

[0457] Example 17: 4-(but-2-ynoyl)-l-(5-methylthiophen-2-yl)piperazin-2-one (XS165-127).

[0458] Example 17 was synthesized following similar procedure for preparing example 1. Colorless oil, 14% yield. 'H NMR (400 MHz, Chloroform-c / ) 5 6.55 (q, J= 3.6, 3.2 Hz, 1H), 6.47 (dt, J = 8.8, 2.8 Hz, 1H), 4.59 (d, J = 2.1 Hz, 1H), 4.43 (d, J= 2.0 Hz, 1H), 4.18 - 4.07 (m, 1H), 4.04 - 3.93 (m, 1H), 3.91 - 3.84 (m, 1H), 3.84 - 3.75 (m, 1H), 2.42 (d, J= 2.4 Hz, 3H), 2.04 (dd, J= 5.0, 2.1 Hz, 3H). MS (ESI) [M+H]+=263.1.

[0459] Scheme 4. Synthesis of example 18 Example 18: 4-(2-fluoroacryloyl)-l-(5-methylthiophen-2-yl)piperazin-2-one (XS165-106). To a solution of 2-bromo-5-methylthiophene (50.0 mg, 0.28 mmol) dissolved in dioxane (1 mL),

[0460] N,N’ -dimethyl ethylenediamine ( 9 pL, 0.0846 mmol, 0.3 eq), K2CO3( 116.9 mg, 0.846 mmol, 3.0 eq), Cui ( 5.4 mg, 0.028 mmol, O.leq ) followed by tert'Z-butyl 3 -oxopiperazine- 1 -carboxylate (84.7 mg, 0.42 mmol, 1.5 eq). The reaction mixture was stirred at 100 °C under nitrogen atmosphere overnight. After cooling down to rt, resulting crude mixtures were purified via silica gel column chromatography to yield intermediate 1.

[0461] To a solution of intermediate 1 (40 mg, 0.135 mmol) dissolved in DCM (0.5 mL), and the TFA(

[0462] O.5 mL) were added. The reaction mixture stirred at rt 1 h. Excess TFA was removed, resulting crude product was re-dissovled in DMF (1 mL), EDCI (58.8 mg, 0.306 mmol, 2.0 eq) and HOAT (41.7 mg, 0.306 mmol, 2.0 eq) were added. At rt the 2-fluoroacrylic acid (13.8 mg, 0.153 mmol, 1.0 eq.) was added, the reaction mixture stirred at rt for 2 h. Resulting crude mixtures were purified via prep-HPLC to yield title compound (White solid, 39% yield) 'H NMR (400 MHz, MethanoL d4) 8 6.67 - 6.62 (m, 1H), 6.60 (d, J= 4.0 Hz, 1H), 5.50 - 5.35 (m, 1H), 5.35 - 5.29 (m, 1H), 4.44 (s, 2H), 4.05 (t, J = 5.5 Hz, 2H), 3.93 (t, J = 5.5 Hz, 2H), 2.41 (s, 3H). MS (ESI) [2(M-1)+H]’ =535.4.

[0463] Example 19: 4-(2-fluoroacryloyl)-l-(5-methylthiazol-2-yl)piperazin-2-one (XS165-112).

[0464] Example 19 was synthesized following similar procedure for preparing example 18. White solid, 40% yield.1H NMR (400 MHz, Methanol-d4) 8 7.15 (s, 1H), 5.44 - 5.29 (m, 1H), 5.29 - 5.24 (m,

[0465] 1H), 4.47 (s, 2H), 4.19 (d, J = 5.7 Hz, 2H), 4.00 (t, J= 5.7 Hz, 2H), 2.36 (d, J = 2.0 Hz, 3H). MS

[0466] (ESI) [M+H]+=309.1. Example 20: 4-(2-fluoroacryloyl)-l-(m-tolyl)piperazin-2-one (XS165-97). Example 20 was synthesized following similar procedure for preparing example 18. White solid, 56% yield. 'H NMR (400 MHz, Chloroform-; / ) 5 7.34 - 7.28 (m, 1H), 7.15 - 7.09 (m, 2H), 7.06 (d, J= 8.0 Hz,

[0467] 1H), 5.57 - 5.35 (m, 1H), 5.34 - 5.14 (m, 1H), 4.44 (s, 2H), 3.98 (t, J= 5.7 Hz, 2H), 3.78 (t, J = 4.8 Hz, 2H), 2.37 (s, 3H). MS (ESI) [M+H]+=263.7.

[0468] Example 21: 2-fluoro-l-(3-(6-methylpyridin-2-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)prop-2- en-l-one (XS165-118). Example 21 was synthesized following similar procedure for preparing example 18. White solid, 45% yield. 'H NMR (400 MHz, Methanol-^) 5 8.01 - 7.87 (m, 1H), 7.17 (d, J = 9.2 Hz, 1H), 6.90 (d, J = 7.3 Hz, 1H), 5.49 (dt, J = 47.5, 2.9 Hz, 1H), 5.31 (dt, J = 16.9, 2.9 Hz, 1H), 4.87 (s, 16H), 4.02 (d, J= 12.1 Hz, 2H), 3.51 - 3.39 (m, 2H), 2.58 (d, J= 2.1 Hz, 3H), 2.08 (s, 2H), 1.93 (d, J= 8.7 Hz, 2H). MS (ESI) [M+H]+=276.2.

[0469] Example 22: 4-(2-fluoroacryloyl)-l-(6-methylpyridin-2-yl)piperazin-2-one (XS165-110).

[0470] Example 22 was synthesized following similar procedure for preparing example 18. White solid, 49% yield. 'H NMR (400 MHz, Methanol-d4) 5 7.80 (td, J = 8.0, 1.8 Hz, 1H), 7.59 (d, J= 8.1 Hz,

[0471] 1H), 7.20 (dd, J= 7.6, 2.1 Hz, 1H), 5.51 - 5.25 (m, 2H), 4.42 (s, 2H), 4.16 - 4.05 (m, 2H), 3.99 (s, 2H), 2.52 (d, J= 2.1 Hz, 3H). MS (ESI) [M+H]+=264.1.

[0472] Example 23: 4-(2-chloroacetyl)-l-(5-methylthiophen-2-yl)piperazin-2-one (XS165-119).

[0473] Example 23 was synthesized following similar procedure for preparing example 1. White solid, 32% yield. *HNMR (400 MHz, Methanol-d4) 8 6.64 (d, J= 3.9 Hz, 1H), 6.60 (d, J= 3.9 Hz, 1H), 4.48 - 4.30 (m, 4H), 4.08 - 3.84 (m, 4H), 2.41 (s, 3H). MS (ESI) [M+H]+=273.1. hloroacetyl)-l-(5-methylthiazol-2-yl)piperazin-2-one (XS165-123) . esized following similar procedure for preparing example 1. White solid, 400 MHz, Chloroform-^ / ) 8 7.16 (s, 1H), 4.48 (d, J= 6.3 Hz, 2H), 4.34 (d, d, . / = 5.4 Hz, 1H), 4.12 (d, , / = 8.6 Hz, 2H), 4.04 - 3.88 (m, 2H), 2.41 (s, =273.4. luoroacetyl)-l-(5-methylthiophen-2-yl)piperazin-2-one (XS165-126). esized following similar procedure for preparing example 18. White solid, 00 MHz, Chloroform-d / ) 8 6.56 (d, J = 3.7 Hz, 1H), 6.49 (d, J = 5.5 Hz, Hz, 1H), 4.99 (d, J= 10.2 Hz, 1H), 4.40 (d, J= 34.5 Hz, 2H), 4.09 - 3.77 1 Hz, 3H). MS (ESI) [M+H]+=257.1.

[0474] Example 26: 4-(2-chloroacetyl)-l-(6-methylpyridin-2-yl)piperazin-2-one (XS165-120).

[0475] Example 26 was synthesized following similar procedure for preparing example 18. White solid, 35% yield. 'H NMR (400 MHz, Chloroform-d / ) δ 7.72 (dd, J= 23.9, 8.2 Hz, 1H), 7.61 (t, J= 7.9 Hz, 1H), 6.99 (t, J= 6.7 Hz, 1H), 4.40 (d, J= 9.7 Hz, 2H), 4.30 - 4.23 (m, 1H), 4.17 (d, J= 5.7 Hz, 1H), 4.12 (d, J= 2.9 Hz, 2H), 3.97 - 3.84 (m, 2H), 2.50 (s, 3H). MS (ESI) [M+H]+=268.1.

[0476] Example 27: 2-chloro-l-(3-(6-methylpyridin-2-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)ethan-l- one (XS165-121). Example 27 was synthesized following similar procedure for preparing example 18. White solid, 47% yield. 'H NMR (400 MHz, Chloroform-tT) 8 7.44 - 7.30 (m, 1H), 6.52 (d, J = 7.3 Hz, 1H), 6.35 (d, J= 8.5 Hz, 1H), 4.82 (d, J = 6.4 Hz, 1H), 4.37 (d, J= 6.5 Hz, 1H), 4.20 (d, J= 12.2 Hz, 1H), 4.13 - 4.01 (m, 2H), 3.92 (d, J= 12.0 Hz, 1H), 3.08 (dd, J= 12.1, 2.9 Hz, 2H), 2.39 (s, 3H), 2.10 - 1.77 (m, 4H). MS (ESI) [M+H|+=280.1.

[0477] Scheme 5. Synthesis of example 28

[0478] Example 28: (£)-4-(4-(dimethylamino)but-2-enoyl)-l-(5-methylfuran-2-yl)piperazin-2-one (XS165-100). To a solution of 2-bromo-5-methylfuran(50.0 mg, 0.28 mmol) dissolved in dioxane (1 mL), N,N’ -dimethylethylenediamine ( 9 μL, 0.0846 mmol, 0.3 eq), K2CO3 ( 116.9 mg, 0.846 mmol, 3.0 eq), Cui ( 5.4 mg, 0.028 mmol, O. leq ) followed by tert -butyl 3 -oxopiperazine- 1- carboxylate (84.7 mg, 0.42 mmol, 1.5 eq). The reaction mixture was stirred at 100 °C under nitrogen atmosphere overnight. After cooling down to rt, resulting crude mixtures were purified via silica gel column chromatography to yield intermediate 5.

[0479] To a solution of intermediate 1 (30 mg, 0.107 mmol) dissolved in DCM (0.5 mL), and the TFA( 0.5 mL) were added. The reaction mixture stirred at rt 1 h. Excess TFA was removed, resulting crude product was re-dissovled in DMF (1 mL), EDCI (41.1 mg, 0.214 mmol, 2.0 eq) and HOAT (29.2 mg, 0.214 mmol, 2.0 eq) were added. At rt the (£)-4-(dimethylamino)but-2-enoic acid (17.7 mg, 0.107 mmol, 1.0 eq.) was added, the reaction mixture stirred at rt for 2 h. Resulting crude mixtures were purified via prep-HPLC to yield title compound (White solid, 31% yield) 'H NMR (400 MHz, Methanol-d4) δ 7.02 - 6.87 (m, 1H), 6.78 (q, J= 9.6, 8.5 Hz, 1H), 6.22 (t, J= 2.7 Hz, 1H), 6.08 - 6.00 (m, 1H), 4.44 (d, J= 28.2 Hz, 2H), 4.05 - 3.96 (m, 4H), 3.93 - 3.83 (m, 2H), 2.93 (d, J= 2.1 Hz, 6H), 2.26 (d, J= 1.8 Hz, 3H). MS (ESI) [M+H]+=292.2.

[0480] Example 29: (£)-4-(4-(dimethylamino)but-2-enoyl)-l-( / / i-tolyl)piperazin-2-one (XS165-99).

[0481] Example 29 was synthesized following similar procedure for preparing example 28. White solid, 54% yield.1HMR C400Hz, Methanol-^) 87.36 - 7.24 (m, 1H), 7.12 (d, J= 5.7 Hz, 2H), 7.08 (d, 7 = 8.0 Hz, 1H), 6.92 (dd, 7= 22.6, 15.1 Hz, 1H), 6.75 (dt, 7= 14.8, 7.1 Hz, 1H), 4.40 (d, J = 26.9 Hz, 2H), 4.00 (p, J= 5.2 Hz, 2H), 3.94 (d, J= 7.2 Hz, 2H), 3.88 - 3.72 (m, 2H), 2.88 (d, J = 1.8 Hz, 6H), 2.33 (d, J= 1.8 Hz, 3H). MS (ESI) [M+H]+=302.1.

[0482] Example 31: (£)-4-(4-(dimethylamino)but-2-enoyl)-l-(6-methylpyridin-2-yl)piperazin-2-one (XS165-109). Example 31 was synthesized following similar procedure for preparing example 28. White solid, 47% yield. ’H NMR (400 MHz, Methanol-d4) 8 7.74 (t, 7 = 7.9 Hz, 1H), 7.59 (t, 7 = 9.7 Hz, 1H), 7.16 (d, J = 7.5 Hz, 1H), 6.95 (t, J = 16.4 Hz, 1H), 6.85 - 6.72 (m, 1H), 4.48 (d, 7 = 27.1 Hz, 2H), 4.16 (d, J= 5.6 Hz, 1H), 4.09 (t, J= 5.0 Hz, 1H), 4.06 - 3.93 (m, 4H), 2.93 (d, J = 2.0 Hz, 6H), 2.52 (d, J= 2. 1 Hz, 3H). MS (ESI) [M+H]+=303.2.

[0483] Example 34: (E')-l-(5-methylthiophen-2-yl)-4-(4-(pyrrolidin-l-yl)but-2-enoyl)piperazin-2- one (XS165-154). Example 34 was synthesized following similar procedure for preparing example 28. White solid, 58% yield. 'H NMR (400 MHz, Methanol-^) 8 6.90 (dd, J = 36.2, 15.2 Hz, 1H), 6.81 - 6.68 (m, 1H), 6.65 - 6.44 (m, 2H), 4.43 (d, J= 30.7 Hz, 2H), 4.00 (d, J= 7.1 Hz, 4H), 3.94 - 3.78 (m, 2H), 3.62 (s, 2H), 3.12 (s, 2H), 2.37 (d, J= 2.3 Hz, 3H), 2.08 (d, J= 48.5 Hz, 4H). MS (ESI) [M+H]+=334.2.

[0484] Example 35: (E)-l-(5-methylthiophen-2-yl)-4-(4-(piperidin-l-yl)but-2-enoyl)piperazin-2-one (XS165-170). Example 35 was synthesized following similar procedure for preparing example 28. White solid, 55% yield.1HNMR(400 MHz, Methanol-d4) δ 6.91 (dd, J= 34.6, 15.2 Hz, 1H), 6.82 - 6.67 (m, 1H), 6.57 (dt, J= 11.4, 3.5 Hz, 2H), 4.43 (d, J= 29.1 Hz, 2H), 4.02 (q, J= 8.4, 6.8 Hz,

[0485] 2H), 3.88 (dd, J= 25.4, 6.4 Hz, 4H), 3.51 (d, J= 12.4 Hz, 2H), 2.94 (t, J= 12.5 Hz, 2H), 2.37 (d,

[0486] J = 2.3 Hz, 3H), 1.95 (d, J= 14.4 Hz, 2H), 1.88 - 1.63 (m, 3H), 1.59 - 1.40 (m, 1H). MS (ESI) [M+H]+=348.3.

[0487] Example 37: (E')-4-(dimethylamino)-l-(4-(5-methylthiophen-2-yl)piperazin-l-yl)but-2-en-l- one (XS165-172). Example 37 was synthesized following similar procedure for preparing example 28. White solid, 33% yield. 'H NMR (400 MHz, Methanol-^) 8 7.00 - 6.72 (m, 2H), 6.73 - 6.30 (m, 2H), 4.12 - 3.84 (m, 8H), 3.76 - 3.69 (m, 2H), 2.89 - 2.85 (m, 9H). MS (ESI) [M+H]+=294.4.

[0488] Scheme 6. Synthesis of example 38

[0489] Example 38: (£)-4-(dimethylamino)-l-(4-(5-methylthiophen-2-yl)piperidin-l-yl)but-2-en-l- one (XS165-169). ). To a solution of the 2-bromo-5-methylthiophene (177.1 mg, 1 mmol) dissolved in dioxane / H2O = 4: 1 (3 mL), K3PO4(424.6 mg, 2 mmol, 2.0 eq), Pd (PPh3)2C12(70.2 mg, 0.1 mmol, 0.1 eq), followed by tert-butyl 4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6- dihydropyridine-l(277)-carboxylate (618.4 mg, 2.0 mmol, 2.0 eq). The reaction mixture was stirred at 90 °C under nitrogen atmosphere overnight. After cooling down to rt, resulting crude mixtures were purified via silica gel column chromatography to yield intermediate 5.

[0490] To a solution of intermediate 5 (200 mg, 0.72 mmol) dissolved in MeOH (3 mL), and the Pd / C( 20 mg) were added. The reaction mixture stirred at hydrogen atmosphere. MeOH was removed, resulting crude product was re-dissovled in DCM (1 mL), TFA (1 mL) was added. The reaction mixture stirred at rt for 1 h. Excess TFA was removed, resulting crude product was re-dissovled in DMF (1 mL), EDCI (272 mg, 1.42 mmol, 2.0 eq) and HOAT (187.4 mg, 1.42 mmol, 2.0 eq) were added. At rt the (E)-4-(dimethylamino)but-2-enoic acid (117.6 mg, 0.71 mmol, 1.0 eq.) was added, the reaction mixture stirred at rt for 2 h. Resulting crude mixtures were purified via prep- HPLC to yield title compound (White solid, 43% yield)1H NMR (400 MHz, Methanol-d4) 5 6.95 (d, J= 15.1 Hz, 1H), 6.72 - 6.58 (m, 2H), 6.55 (d, J= 2.8 Hz, 1H), 4.59 (d, J= 13.4 Hz, 1H), 4.13 (d, J= 13.9 Hz, 1H), 3.92 (dd, J= 7.3, 2.2 Hz, 2H), 3.24 (d, J= 13.1 Hz, 1H), 3.14 - 3.00 (m, 1H), 2.95 - 2.76 (m, 7H), 2.38 (d, J= 2.3 Hz, 3H), 2.13 - 1.96 (m, 2H), 1.74 - 1.42 (m, 2H). MS (ESI) [M+H]+=293.4.

[0491] Example 39: (E)-4,4-dimethyl-2-(4-(5-methylthiophen-2-yl)-3-oxopiperazine-l- carbonyl)pent-2-enenitrile (XS165-177). Example 39 was synthesized following similar

[0492] I l l procedure for preparing example 28. White solid, 47% yield.1H NMR (400 MHz, Methanol-^) 8 7.12 - 6.97 (m, 1H), 6.67 - 6.43 (m, 2H), 4.38 (s, 2H), 4.08 - 3.76 (m, 4H), 2.39 (s, 3H), 1.32 Example 46: (E)-4-(4-(dimethylamino)but-2-enoyl)-l -(3-methylthiophen-2-yl)piperazin-2- one (XS175-68). Example 46 was synthesized following similar procedure for preparing example 28. White solid, 51% yield. 'H NMR (400 MHz, Methanol-d4) δ 7.23 (d, J= 5.3 Hz, 1H), 7.03 - 6.87 (m, 1H), 6.84 (d, J= 5.4 Hz, 1H), 6.77 (dt, J= 14.8, 7.2 Hz, 1H), 4.46 (d, J= 30.5 Hz, 2H), 4.04 (q, J= 5.8 Hz, 2H), 4.00 - 3.87 (m, 2H), 3.82 - 3.64 (m, 2H), 2.90 (s, 6H), 2.06 (s, 3H). MS

[0493] Example 47 : (£)-4-(4-(dimethylamino)but-2-enoyl)-l-(4,5-dimethylthiophen-2-yl)piperazin- 2-one (XS175-70). Example 47 was synthesized following similar procedure for preparing example 28. White solid, 53% yield. 'H NMR (400 MHz, Methanol-d4) 8 6.93 (dd, J = 36.6, 15.2 Hz, 1H), 6.76 (dt, J= 15.0, 7.4 Hz, 1H), 6.50 (s, 1H), 4.43 (d, J= 29.5 Hz, 2H), 4.03 (s, 2H), 3.95 (d, J= 7.3 Hz, 2H), 3.93 - 3.77 (m, 2H), 2.90 (s, 6H), 2.24 (s, 3H), 2.07 (s, 3H). MS (ESI) [M+H]- =322.2.

[0494] Example 48: (E)-l-(4-(dimethylamino)but-2-enoyl)-W-(5-methylthiophen-2-yl)azetidine-3- carboxamide (XS175-71) . Example 48 was synthesized following similar procedure for preparing example 28. White solid, 24% yield. 'H NMR (400 MHz, Methanol-d4) 8 6.80 - 6.62 (m, 1H), 6.53 - 6.37 (m, 3H), 4.54 - 4.39 (m, 2H), 4.29 - 4.20 (m, 1H), 4.19 - 4.11 (m, 1H), 3.95 - 3.85 (m, 2H), 3.63 - 3.51 (m, 1H), 2.87 (s, 6H), 2.35 (s, 3H). MS (ESI) [M+H]+=308.3

[0495] Example 49: (E)-4-(dimethylamino)-JV-(2-((5-methylthiophen-2-yl)amino)-2-oxoethyl)but-2- enamide (XS175-76). Example 49 was synthesized following similar procedure for preparing example 28. White solid, 20% yield. 'H NMR (400 MHz, Methanol-d4) 8 6.74 - 6.56 (m, 1H), 6.46 - 6.32 (m, 3H), 4.04 - 3.95 (m, 2H), 3.90 - 3.81 (m, 2H), 2.81 (s, 6H), 2.28 (s, 3H). MS (ESI) [M+H]+=282.5.

[0496] Example 52: l-(2-chloroacetyl)-JV-(5-methylthiophen-2-yl)azetidine-3-carboxamide (XS175- 126). Example 52 was synthesized following similar procedure for preparing example 28. White solid, 33% yield.JH NMR (400 MHz, Methanol-d4) 8 6.56 - 6.37 (m, 2H), 4.55 - 4.40 (m, 2H), 4.29 - 4.12 (m, 2H), 4.09 - 4.00 (m, 2H), 3.66 - 3.53 (m, 1H), 2.38 (s, 3H). MS (ESI) [M+H]+=273.1.

[0497] Scheme 7. Synthesis of example 53

[0498] Example 53: (E)-l-(4-(dimethylamino)but-2-enoyl)-A-(5-methylthiophen-2-yl)piperidine-4- carboxamide (XS175-132). ). To a solution of the 5-methylthiophen-2-amine (113.2 mg, 1 mmol) dissolved in DMF (2 mL), HATU (418.2 mg, 1.1 mmol, 1.1 eq), DIPEA (0.18 mL, 2.0 mmol, 2.0 eq), followed by l-(terZ-butoxycarbonyl)piperidine-4-carboxylic acid (229.3 mg, 1.0 mmol, 1.0 eq). The reaction mixture was stirred at rt 3 h. The resulting crude mixtures were purified via silica gel column chromatography. The resulting crude product was re-dissovled in DCM (1 mL), TFA (1 mL) was added. The reaction mixture stirred at rt for 1 h. Excess TFA was removed, resulting crude product was were purified via silica gel column chromatography to yield intermediate 7.

[0499] To a solution of intermediate 7 (25 mg, 0.116 mmol) dissolved in DMF (1 mL), HATU (50.9 mg, 0.14 mmol, 1.2 eq), DIPEA (0.043 mL, 0.24 mmol, 2.0 eq), followed by (E)-4-

[0500] (dimethylamino)but-2-enoic acid (18.5 mg, 0. 116 mmol, 1.0 eq) the reaction mixture stirred at rt for 2 h. Resulting crude mixtures were purified via prep-HPLC to yield title compound (White solid, 43% yield)1H NMR (400 MHz, Methanol-d)48 6.88 (d, J= 15.1 Hz, 1H), 6.65 - 6.50 (m, 1H), 6.41 (s, 2H), 4.49 (d, J = 12.9 Hz, 1H), 4.07 (d, J= 13.5 Hz, 1H), 3.85 (d, J= 6.7 Hz, 2H),

[0501] 3.16 (t, J = 12.9 Hz, 1H), 2.82 (s, 7H), 2.59 (t, J = 9.8 Hz, 1H), 2.28 (s, 3H), 1.84 (d, J= 12.4 Hz, 2H), 1.61 (p, J= 13.0 Hz, 2H). MS (ESI) [M+H]+=336.2.

[0502] Example 54: (E)-l-(4-(dimethylamino)but-2-enoyl)- / V-(5-methylthiophen-2-yl)pyrrolidine-

[0503] 3-carboxamide (XS175-133). Example 54 was synthesized following similar procedure for preparing example 53. White solid, 29% yield. 'H NMR (400 MHz, Methanol-d4) 8 6.73 - 6.56 (m, 2H), 6.40 (s, 2H), 3.85 (s, 2H), 3.83 - 3.37 (m, 4H), 3.20 - 3.04 (m, 1H), 2.80 (s, 6H), 2.27 (s, 3H), 2.25 - 1.94 (m, 2H). MS (ESI) [M+H]+=322.

[0504] Example 56: (£)-l-(4-(dimethylamino)but-2-enoyl)-7V-(5-isopropylthiophen-2-yl)azetidine-

[0505] 3-carboxamide (XS175-143). Example 56 was synthesized following similar procedure for preparing example 53. White solid, 37% yield. 'H NMR (400 MHz, Methanol-d4) 8 6.75 - 6.59 (m, 1H), 6.57 - 6.40 (m, 3H), 4.56 - 4.36 (m, 2H), 4.24 (t, J= 9.4 Hz, 1H), 4.19 - 4.09 (m, 1H), 3.90 (d, J= 6.7 Hz, 2H), 3.63 - 3.50 (m, 1H), 3.03 (h, J= 6.5, 5.6 Hz, 1H), 2.86 (s, 6H), 1.25 (d, J= 6.5 Hz, 6H). MS (ESI) [M+H]+=336.2.

[0506] Example 57: (£)-l-(4-(dimethylamino)but-2-enoyl)- / V-(5-methylthiophen-2-yl)piperidine-2- carboxamide (XS175-148). Example 57 was synthesized following similar procedure for preparing example 53. White solid, 25% yield. 'H NMR (400 MHz, Methanol-d4) δ 7.07 - 6.79 (m, 1H), 6.72 - 6.43 (m, 3H), 5.23 (s, 1H), 4.10 - 3.82 (m, 3H), 3.61 - 3.44 (m, 1H), 2.89 (s, 6H), 2.36 (s, 3H), 2.29 - 2.14 (m, 1H), 1.87 - 1.64 (m, 3H), 1.63 - 1.41 (m, 2H). MS (ESI) [M+Hf =336.2.

[0507] Example 58: (E)-2-(4-(dimethylamino)but-2-enoyl)-JV-(5-methylthiophen-2-yl)-2- azaspiro[3.3]heptane-6-carboxamide (XS175-149). Example 58 was synthesized following similar procedure for preparing example 53. White solid, 16% yield. 'H NMR (400 MHz, Methanol-^) 5 6.70 - 6.53 (m, 1H), 6.49 - 6.28 (m, 3H), 4.39 - 4.18 (m, 2H), 4.09 - 3.95 (m,

[0508] 2H), 3.94 - 3.80 (m, 2H), 3.13 - 2.97 (m, 1H), 2.84 (s, 6H), 2.44 (d, J= 7.9 Hz, 4H), 2.31 (s, 3H).

[0509] MS (ESI) [M+H]+=348.3. Example 61 : (E)-4-(dimethylamino)-2V-methyl-iV-(3-(methyl(5-methylthiophen-2-yl)amino)- 3-oxopropyl)but-2-enamide (XS175-160) . Example 61 was synthesized following similar procedure for preparing example 28. White solid, 23% yield.JH NMR (400 MHz, Methanol-d4) 8 6.86 - 6.70 (m, 2H), 6.64 (s, 1H), 6.40 (d, J= 15.4 Hz, 1H), 4. 12 (d, J= 7.2 Hz, 2H), 3.52 - 3.43 (m, 2H), 3.23 (s, 3H), 3.15 (s, 9H), 2.45 (s, 3H). MS (ESI) [M+H]+=324.2.

[0510] Example 62: (E)-l-(4-(dimethylamino)but-2-enoyl)-JV-(4-methylthiophen-2-yl)piperidine-4- carboxamide (XS175-173). Example 62 was synthesized following similar procedure for preparing example 28. White solid, 46% yield. 'H NMR (400 MHz, Methanol-6 / 4) δ 6.93 (d, J = 15.1 Hz, 1H), 6.62 (dt, J= 14.6, 6.8 Hz, 1H), 6.51 (s, 1H), 4.54 (d, J= 12.9 Hz, 1H), 4.12 (d, J = 13.4 Hz, 1H), 3.90 (d, .7= 6.8 Hz, 2H), 3.21 (t, ,7= 12.9 Hz, 1H), 2.87 (s, 7H), 2.70 - 2.58 (m, 1H), 2.13 (s, 3H), 1.89 (d, J= 12.5 Hz, 2H), 1.78 - 1.52 (m, 2H). MS (ESI) [M+H]+=336.2.

[0511] Example 63: (E)-l-(4-(dimethylamino)but-2-enoyl)-JV,7V-bis(5-methylthiophen-2- yl)piperidine-4-carboxamide (XS175-174). Example 63 was synthesized following similar procedure for preparing example 28. White solid, 13% yield. 'H NMR (400 MHz, Methanol-6 / 4) 8 7.17 - 6.88 (m, 2H), 6.77 (s, 1H), 6.70 - 6.58 (m, 1H), 6.47 (s, 1H), 6.21 (s, 1H), 4.53 (d, J = 13.1 Hz, 1H), 4.09 (d, .7= 13.5 Hz, 1H), 3.93 (d, .7= 7.0 Hz, 2H), 3.06 (t, ,7= 12.8 Hz, 1H), 3.01 - 2.94 (m, 1H), 2.94 - 2.86 (m, 6H), 2.66 (t, J= 12.9 Hz, 1H), 2.45 (d, J= 49.1 Hz, 6H), 1.89 (d, J = 13.0 Hz, 2H), 1.82 - 1.59 (m, 2H). MS (ESI) [M+H]+=432.2.

[0512] Example 64: (E)-l-(4-(dimethylamino)but-2-enoyl)-W^V-bis(5-isopropylthiophen-2- yl)piperidine-4-carboxamide (XS175-175). Example 64 was synthesized following similar procedure for preparing example 28. White solid, 19% yield. 'H NMR (400 MHz, Methanol-d4) 8 7.18 - 6.89 (m, 2H), 6.81 (d, J= 18.9 Hz, 1H), 6.70 - 6.60 (m, 1H), 6.60 - 6.38 (m, 1H), 6.26 (s, 1H), 4.54 (d, J= 13.4 Hz, 1H), 4.10 (d, J= 13.9 Hz, 1H), 3.98 - 3.86 (m, 2H), 3.29 - 3.02 (m, 4H), 2.92 (s, 7H), 2.76 - 2.57 (m, 1H), 1.91 (d, J = 13.3 Hz, 2H), 1.84 - 1.64 (m, 2H), 1.34 (s,

[0513] Example 65: (£)-l-(4-(dimethylamino)but-2-enoyl)-Af^V-bis(5-isopropylthiophen-2- yl)pyrrolidine-3-carboxamide (XS175-176). Example 65 was synthesized following similar procedure for preparing example 28. White solid, 20% yield. 'H NMR (400 MHz, Methanol-d4) 8 7.14 - 6.94 (m, 1H), 6.83 (s, 1H), 6.73 (s, 2H), 6.64 - 6.46 (m, 1H), 6.28 (s, 1H), 3.95 (s, 2H), 3.90 - 3.72 (m, 2H), 3.66 (t, J= 7.5 Hz, 1H), 3.57 - 3.37 (m, 2H), 3.27 - 3.02 (m, 2H), 2.92 (s, 6H), 2.40 - 2.09 (m, 2H), 1.38 - 1.28 (m, 12H). MS (ESI) [M+H]+=474.3.

[0514] Example 66: (E)-l-(4-(dimethylamino)but-2-enoyl)- / V-(5-isopropylthiophen-2-yl)piperidine- 4-carboxamide (XS175-178). Example 66 was synthesized following similar procedure for preparing example 28. White solid, 49% yield. 'H NMR (400 MHz, Methanol-d4) 8 6.95 (d, J = 15.0 Hz, 1H), 6.71 - 6.57 (m, 1H), 6.51 (d, J= 9.7 Hz, 2H), 4.57 (d, J = 12.9 Hz, 1H), 4.14 (d, J = 13.4 Hz, 1H), 3.93 (d, J= 6.8 Hz, 2H), 3.23 (d, J= 12.9 Hz, 1H), 3.11 - 2.99 (m, 1H), 2.90 (s, 7H), 2.74 -2.56 (m, 1H), 1.92 (d, J= 13.5 Hz, 2H), 1.80 - 1.57 (m, 2H). MS (ESI) [M+H]+=364.7.

[0515] Example 67: (E)-l-(benzo[Z>]thiophen-2-yl)-4-(4-(dimethylamino)but-2-enoyl)piperazin-2- one (XS175-179). Example 67 was synthesized following similar procedure for preparing example 28. White solid, 46% yield. 'H NMR (400 MHz, Methanol-d)48 7.83 - 7.60 (m, 2H), 7.40 - 7.20 (m, 2H), 7.11 - 6.91 (m, 2H), 6.79 (d, J= 14.6 Hz, 1H), 4.54 (d, J= 30.3 Hz, 2H), 4.23 - 3.86 (m, 6H), 2.93 (s, 6H). MS (ESI) [M+H]+=344.4.

[0516] Example 70: (E)-4-(dimethylamino)-l-(6-(5-methylthiophen-2-yl)-2,6- diazaspiro[3.3]heptan-2-yl)but-2-en-l-one (XS186-3). Example 70 was synthesized following similar procedure for preparing example 28. White solid, 23% yield. 'H NMR (400 MHz, Methanol- r) 8 6.69 (s, 2H), 6.55 - 6.32 (m, 2H), 4.50 (s, 2H), 4.22 (d, J = 3.2 Hz, 2H), 4.01 - 3.91 (m, 6H), 2.90 - 2.88 (m, 6H), 2.32 (d, J= 3.4 Hz, 3H). MS (ESI) [M+H]+=306.2.

[0517] Example 73: l-(2-chloroacetyl)-AL(5-isopropylthiophen-2-yl)piperidine-4-carboxamide (XS185-6). Example 73 was synthesized following similar procedure for preparing example 28. White solid, 30% yield. ’H NMR (400 MHz, Methanol-d)48 6.59 - 6.45 (m, 2H), 4.60 - 4.43 (m, 1H), 4.36 - 4.18 (m, 2H), 4.08 - 3.95 (m, 1H), 3.25 (t, J= 13.1 Hz, 1H), 3.14 - 3.00 (m, 1H), 2.83 (t, J= 12.7 Hz, 1H), 2.71 - 2.58 (m, 1H), 2.02 - 1.77 (m, 3H), 1.76 - 1.62 (m, 1H), 1.34 - 1.23 (m, 6H). MS (ESI) [M+H]+=329.1.

[0518] Scheme 8. Synthesis of example 74

[0519]

[0520] Example 74

[0521] Example 74: (£)-4-(4-(dimethylamino)but-2-enoyl)-l-(5-(piperidin-4-yl)thiophen-2- yl)piperazin-2-one (XS185-24)To a solution of tert-butyl 4-(5-brom othi ophen-2 -yl)piperidine-l- carboxylate (123.0 mg, 0.35 mmol) dissolved in dioxane (2 mL), N,N’-dimethylethylenediamine ( 9 pL, 0.105 mmol, 0.3 eq), K2CO3( 147.2 mg, 1.05 mmol, 3.0 eq), Cui ( 6.8 mg, 0.035 mmol, O.leq ) followed by benzyl 3 -oxopiperazine- 1 -carboxylate (125.1 mg, 0.54 mmol, 1.5 eq). The reaction mixture was stirred at 100 °C under nitrogen atmosphere overnight. After cooling down to rt, resulting crude mixtures were purified via silica gel column then the product dissolved in MeOH (3 mL), and the Pd / C( 20 mg) were added. The reaction mixture stirred at hydrogen atmosphere. MeOH was removed then purified via silica gel column chromatography to yield intermediate 8.

[0522] To a solution of intermediate 8 was dissovled in DMF (1 mL), HATU (59.3 mg, 0.144 mmol, 1.1 eq) and DIPEA (0.1 mL, 2.0 eq) were added. At rt the (E)-4-(dimethylamino)but-2-enoic acid (17.7 mg, 0.107 mmol, 1.0 eq.) was added, the reaction mixture stirred at rt for 2 h. Resulting crude mixtures were purified via prep-HPLC then re-dissolved in DCM (0.5 mL), and the TFA( 0.5 mL) were added. The reaction mixture stirred at rt 1 h. Excess TFA was removed, resulting crude product was purified via prep-HPLC to yield title compound (White solid, 19% yield) 'H NMR (400 MHz, Methanol-d4) δ 6.95 (dd, J= 38.8, 15.4 Hz, 1H), 6.86 - 6.62 (m, 3H), 4.48 (d, J = 29.9 Hz, 2H), 4.16 - 4.01 (m, 2H), 4.01 - 3.85 (m, 4H), 3.46 (d, ,7= 13.0 Hz, 2H), 3.12 (t, J = 12.5 Hz, 4H), 2.91 (s, 6H), 2.22 (d, J= 14.2 Hz, 2H), 1.89 (q, J = 13.1 Hz, 2H), 1.45 - 1.12 (m, Example 75: (E)-4-(dimethylamino)-l-(3-(5-methylthiophen-2-yl)-3,8- diazabicyclo[3.2.1]octan-8-yl)but-2-en-l-one (XS185-29). Example 75 was synthesized following similar procedure for preparing example 28. White solid, 19% yield. 'H NMR (400 MHz, Methanol-d4) 8 6.94 - 6.74 (m, 3H), 6.36 (s, 1H), 4.78 - 4.40 (m, 3H), 3.99-3.95 (m, 4H), 2.92 (m, 8H), 2.31 (s, 3H), 2.09 - 2.03 (m, 2H), 1.98 (td, J= 14.5, 12.6, 8.6 Hz, 3H). MS (ESI) [M+H]+=320.2.

[0523] Example 76: (JE)-4-(4-(dimethylamino)but-2-enoyl)-l-(lH-indol-3-yl)piperazin-2-one (XS185-43). Example 76 was synthesized following similar procedure for preparing example 28. White solid, 32% yield. 'H NMR (400 MHz, Methanol-t / 4) 8 7.40 (t, J= 9.8 Hz, 2H), 7.32 (d, J = 3.1 Hz, 1H), 7.14 (d, J = 9.3 Hz, 1H), 7.07 (d, J= 8.6 Hz, 1H), 7.02 - 6.87 (m, 1H), 6.81 - 6.64 (m, 1H), 4.64 - 4.38 (m, 2H), 4.09 (t, J= 5.4 Hz, 2H), 4.02 - 3.79 (m, 4H), 2.88 (s, 6H). MS (ESI) [M+H]+=327.2.

[0524] Example 77 : (E)- l-(6-(benzo [b] thiophen-2-yl)-2,6-diazaspiro [3.3] heptan-2-yl)-4-

[0525] (dimethylamino)but-2-en-l-one (XS185-46). Example 77 was synthesized following similar procedure for preparing example 28. White solid, 17% yield.1H NMR (400 MHz, Methanol - h) 8 7.57 (t, J= 6.2 Hz, 1H), 7.43 (t, J= 6.1 Hz, 1H), 7.19 (q, J= 6.8, 6.2 Hz, 1H), 7.05 (t, J= 6.6 Hz, 1H), 6.78 - 6.39 (m, 3H), 4.53 (d, J = 4.7 Hz, 2H), 4.32 - 4.21 (m, 4H), 4.11 (t, J = 3.0 Hz, 3H), 3.94 (t, J= 5.7 Hz, 2H), 2.90 (s, 6H). MS (ESI) [M+H]+=342.4.

[0526] Example 78: (E)-l-(2-(benzo[Z>]thiophen-2-yl)-2,7-diazaspiro[3.5]nonan-7-yl)-4-

[0527] (dimethylamino)but-2-en-l-one (XS185-59). Example 78 was synthesized following similar procedure for preparing example 28. White solid, 19% yield. 'H NMR (400 MHz, Methanol-tL) 8 7.59 - 7.49 (m, 1H), 7.44 - 7.34 (m, 1H), 7.17 (d, J = 9.1 Hz, 1H), 7.06 - 6.89 (m, 2H), 6.76 - 6.49 (m, 2H), 3.94 (t, J= 4.2 Hz, 4H), 3.78 - 3.77 (m, 2H), 3.72 - 3.62 (m, 4H), 2.91 (s, 7H), 1.94 - 1.85 (m, 4H). MS (ESI) [M+H]+=370.2.

[0528] Scheme 9. Synthesis of example 79

[0529] Example 79: (E)-4-(dimethylamino)-l-(4-(5-methylthiophene-2-carbonyl)piperazin-l- yl)but-2-en-l-one (XS185-64). To a solution of 5-methylthiophene-2-carboxylic acid(75 mg, 0.5 mmol, 1.0 eq) was dissovled in DMF (1 mL), HATU (209 mg, 0.55 mmol, 1.1 eq) and DIPEA (0.18 mL, 2.0 eq) were added. At rt the tert-butyl piperazine- 1 -carboxylate (93.3 mg, 0.5 mmol, 1.0 eq.) was added, the reaction mixture stirred at rt for 2 h. Resulting crude mixtures were purified via prep-HPLC then re-dissolved in DCM (0.5 mL), and the TFA( 0.5 mL) were added. The reaction mixture stirred at rt 1 h. Excess TFA was removed, resulting crude product was purified via prep-HPLC to yield intermediate 9.

[0530] To a solution of intermediate 9(20 mg, 0.095 mmol) was dissovled in DMF (1 mL), HATU (39.8 mg, 0.1 mmol, 1.1 eq) and DIPEA (0.1 mL, 2.0 eq) were added. At rt the (£)-4- (dimethylamino)but-2-enoic acid (15.8 mg, 0.095 mmol, 1.0 eq.) was added, the reaction mixture stirred at rt for 2 h. Resulting crude mixtures were purified via prep-HPLC to yield title compound (White solid, 30% yield) 'H NMR (400 MHz, Methanol-d4) 8 7.28 - 7.18 (m, 1H), 7.01 - 6.86 (m, 1H), 6.78 (d, J= 4.5 Hz, 1H), 6.74 - 6.60 (m, 1H), 4.02 - 3.86 (m, 2H), 3.86 - 3.75 (m, 4H), 3.75 - 3.62 (m, 4H), 2.87 (s, 6H), 2.48 (s, 3H). MS (ESI) [M+H]+=322.2.

[0531] Example 80: (E)-4-(dimethylamino)-JV-(l-(5-methylthiophene-2-carbonyl)piperidin-4- yl)but-2-enamide (XS185-65). Example 80 was synthesized following similar procedure for preparing example 79. White solid, 46% yield. 'H NMR (400 MHz, Methanol-r / i) 8 7.11 (t, J = 4.0 Hz, 1H), 6.72 (d, J= 4.8 Hz, 1H), 6.69 - 6.56 (m, 1H), 6.35 - 6.19 (m, 1H), 4.28 (d, J= 13.5 Hz, 2H), 4.06 - 3.89 (m, 1H), 3.84 (t, J= 5.5 Hz, 2H), 3.20 - 3.03 (m, 2H), 2.82 (s, 6H), 2.42 (s, 3H), 1.90 (d, J= 12.9 Hz, 2H), 1.54 - 1.35 (m, 2H). MS (ESI) [M+H]+=336.2.

[0532] Example 82: (E')-4-(dimethylamino)-l-(6-(5-methylthiophene-2-carbonyl)-3,6- diazabicyclo[3.1.1]heptan-3-yl)but-2-en-l-one (XS185-67). Example 82 was synthesized following similar procedure for preparing example 79. White solid, 57% yield. 'H NMR (400 MHz, Methanol-d4) 5 7.48 - 7.31 (m, 1H), 6.88 - 6.73 (m, 2H), 6.73 - 6.60 (m, 1H), 5.12 - 4.89 (m, 1H), 4.71 - 4.42 (m, 1H), 4.35 - 4.00 (m, 1H), 3.96 - 3.75 (m, 4H), 3.73 - 3.60 (m, 1H), 2.88 - 2.76 (m, 7H), 2.46 (s, 3H), 1.69 - 1.57 (m, 1H). MS (ESI) [M+H]+=334.2.

[0533] Example 83: (E)-4-(dimethylamino)-l-(6-(5-methylthiophene-2-carbonyl)-2,6- diazaspiro[3.3]heptan-2-yl)but-2-en-l-one (XS185-69). Example 83 was synthesized following similar procedure for preparing example 79. White solid, 19% yield. 'H NMR (400 MHz, Methanol-d4) 8 7.27 (d, J= 4.1 Hz, 1H), 6.75 (d, J= 4.6 Hz, 1H), 6.73 - 6.54 (m, 1H), 6.49 - 6.33 (m, 1H), 4.59 (s, 2H), 4.50 - 4.39 (m, 2H), 4.32 - 4.12 (m, 4H), 3.85 (t, J = 5.4 Hz, 2H), 2.81 (s, 6H), 2.42 (s, 3H). MS (ESI) [M+H]+=334.2.

[0534] Example 84: (E)-4-(dimethylamino)-l-(3-(5-methylthiophene-2-carbonyl)-3,8- diazabicyclo[3.2.1]octan-8-yl)but-2-en-l-one (XS185-70). Example 84 was synthesized following similar procedure for preparing example 79. White solid, 46% yield.1H NMR (400

[0535] MHz, Methanol-d4) 8 7.30 - 7.17 (m, 1H), 7.01 - 6.88 (m, 1H), 6.88 - 6.72 (m, 2H), 4.76 (s, 1H), 4.61 (t, J= 4.8 Hz, 1H), 4.38 (d, J= 15.7 Hz, 2H), 3.99 (t, J= 5.1 Hz, 2H), 3.34 (s, 3H), 2.94 (s, 6H), 2.53 (s, 3H), 2.22 - 1.70 (m, 4H). MS (ESI) [M+H]+=348.2.

[0536] Example 85: (E)-4-(dimethylamino)-l-(8-(5-methylthiophene-2-carbonyl)-3,8- diazabicyclo [3.2. l]octan-3-yl)but-2-en- 1-one (XS185-71). Example 85 was synthesized following similar procedure for preparing example 79. White solid, 49% yield. 'H NMR (400 MHz, Methanol-d4) 8 7.38 - 7.29 (m, 1H), 6.94 - 6.82 (m, 1H), 6.80 (d, J= 4.6 Hz, 1H), 6.76 -

[0537] 6.57 (m, 1H), 4.74 (s, 2H), 4.48 - 4.30 (m, 1H), 4.00 - 3.82 (m, 3H), 3.56 - 3.41 (m, 1H), 3.09 - 2.98 (m, 1H), 2.86 (s, 6H), 2.47 (s, 3H), 2.05 - 1.83 (m, 2H), 1.83 - 1.59 (m, 2H). MS (ESI)

[0538] Example 86: (E)-4-(dimethylamino)-l-(3-(5-methylthiophene-2-carbonyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl)but-2-en-l-one (XS185-72). White solid, 54% yield. Example 86 was synthesized following similar procedure for preparing example 79.1H NMR (400 MHz, Methanol-c / 4) 8 7.35 (d, J= 3.8 Hz, 1H), 6.75 (s, 1H), 6.73 - 6.63 (m, 1H), 6.60 - 6.44 (m, 1H), 4.73 (s, 1H), 4.49 (s, 1H), 4.43 - 3.74 (m, 6H), 2.82 (s, 7H), 2.43 (s, 3H), 1.77 - 1.54 (m, 1H). MS (ESI) [M+H]+=334.6.

[0539] Example 88: (£)-l-(benzofuran-2-yl)-4-(4-(dimethylamino)but-2-enoyl)piperazin-2-one (XS185-78) Example 88 was synthesized following similar procedure for preparing example 28. White solid, 36% yield. 'H NMR (400 MHz, Methanol-d4) δ 7.55 (d, J = 5.3 Hz, 1H), 7.44 (s, 1H), 7.32 - 7.20 (m, 2H), 6.91 (d, J= 3.0 Hz, 1H), 6.78 (s, 2H), 4.54 (d, J= 28.2 Hz, 2H), 4.14 (d, J = 37.0 Hz, 4H), 3.99 (d, J = 8.4 Hz, 3H), 3.87 - 3.73 (m, 1H), 2.93 (s, 6H). MS (ESI) [M+Hf =328.2.

[0540] Example 89: (£)-l-(benzo[Z»]thiophen-3-yl)-4-(4-(dimethylamino)but-2-enoyl)piperazin-2- one (XS185-86). Example 89 was synthesized following similar procedure for preparing example 28. White solid, 40% yield. 'H NMR (400 MHz, Methanol^) 6 7.93 (dt, J = 6.2, 3.8 Hz, 1H), 7.78 - 7.58 (m, 2H), 7.52 - 7.37 (m, 2H), 7.02 (t, J= 17.5 Hz, 1H), 6.89 - 6.75 (m, 1H), 4.57 (d, J= 26.5 Hz, 2H), 4.16 (t, J= 5.3 Hz, 2H), 4.06 - 3.97 (m, 2H), 3.95 - 3.80 (m, 2H), 2.94 (s, 6H). MS (ESI) [M+H]+=344.1

[0541] Scheme 10. Synthesis of example 90

[0542] Example 90: 2-((4-(5-methylthiophen-2-yl)-3-oxopiperazin-l-yl)methyl)acrylonitrile

[0543] (XS185-87). To a solution of 2-bromo-5-methylthiophene (50.0 mg, 0.28 mmol) dissolved in dioxane (1 mL), N,N’ -dimethyl ethylenediamine ( 9 μL, 0.0846 mmol, 0.3 eq), K2CO3( 116.9 mg,

[0544] 0.846 mmol, 3.0 eq), Cui ( 5.4 mg, 0.028 mmol, O. leq ) followed by tert rtoxopiperazine- 1-carboxylate (84.7 mg, 0.42 mmol, 1.5 eq). The reaction mixture was stirred at 100 °C under nitrogen atmosphere overnight. After cooling down to rt, resulting crude mixtures were purified via silica gel column chromatography to yield intermediate 1.

[0545] To a solution of intermediate 1 (20 mg, 0.102 mmol) dissolved in DCM (0.5 mL), and the TFA( 0.5 mL) were added. The reaction mixture stirred at rt 1 h. Excess TFA was removed, resulting crude product was re-dissolved in DMF (1 mL), K2CO3 (28.2 mg, 0.2 mmol, 2.0 eq) and KI (16.9 mg, 1.0 eq) were added. At rt the 2-(bromomethyl)acrylonitrile (17.9 mg, 0.124 mmol, 1.2 eq.) was added, the reaction mixture stirred at rt for 5 h. Resulting crude mixtures were purified via prep-HPLC to yield title compound (White solid, 24% yield)XH NMR (400 MHz, Methanol-d4) 8 6.62 (d, J= 14.6 Hz, 2H), 6.36 - 6.13 (m, 2H), 3.92 (d, J= 5.2 Hz, 2H), 3.55 (t, J= 5.1 Hz, 4H), 3.15 (d, J = 5.4 Hz, 2H), 2.41 (s, 3H). MS (ESI) [M+H]+=262.1. Example 92: (E)- / V-(benzo|b ]thiophen-2-yl)-2-(4-(dimethylamino)but-2-enoyl)-2- azaspiro[3.3]heptane-6-carboxamide (XS185-91). Example 92 was synthesized following similar procedure for preparing example 53. White solid, 48% yield. 'H NMR (400 MHz,

[0546] Methanol-^) 8 7.74 - 7.64 (m, 1H), 7.63 - 7.50 (m, 1H), 7.24 (d, J= 8.4 Hz, 1H), 7.18 (d, J= 8.5 Hz, 1H), 6.88 (d, J= 3.6 Hz, 1H), 6.74 - 6.59 (m, 1H), 6.51 - 6.36 (m, 1H), 4.41 - 4.26 (m, 2H), 4.14 - 4.01 (m, 2H), 3.99 - 3.85 (m, 2H), 3.24 - 3.09 (m, 1H), 2.86 (s, 6H), 2.51 (d, J= 5.0 Hz, 4H). MS (ESI) [M+H]+=384.2.

[0547] Example 93: (E')-4-(dimethylamino)-l-(7-(5-methylthiophene-2-carbonyl)-2,7- diazaspiro[3.5]nonan-2-yl)but-2-en-l-one (XS185-96). Example 93 was synthesized following similar procedure for preparing example 79. White solid, 16% yield. 'H NMR (400 MHz, Methanol-^) 8 7.15 (d, J = 3.1 Hz, 1H), 6.78 - 6.73 (m, 1H), 6.73 - 6.63 (m, 1H), 6.46 (d, J = 15.3 Hz, 1H), 4.06 (s, 2H), 3.96 - 3.87 (m, 2H), 3.81 (s, 2H), 3.74 - 3.58 (m, 4H), 2.85 (s, 6H), 2.50 - 2.42 (m, 3H), 1.89 - 1.71 (m, 4H). MS (ESI) [M+H]+=362.2.

[0548] Example 94: (E')-4-(dimethylamino)-l-(6-(5-methylthiophene-2-carbonyl)-2,6- diazaspiro[3.4]octan-2-yl)but-2-en-l-one (XS185-97). Example 94 was synthesized following similar procedure for preparing example 79. White solid, 12% yield. 'H NMR (400 MHz, Methanol-^) 8 7.40 (d, J= 3.8 Hz, 1H), 6.85 - 6.76 (m, 1H), 6.76 - 6.59 (m, 1H), 6.56 - 6.31 (m, 1H), 4.39 - 4.15 (m, 2H), 4.13 - 3.95 (m, 3H), 3.88 (t, J= 10.4 Hz, 3H), 3.69 (d, J= 53.8 Hz, 2H), 2.85 (s, 6H), 2.51 - 2.43 (m, 3H), 2.32 - 2.08 (m, 2H). MS (ESI) [M+H]+=348.2.

[0549] Example 95: (E)-4-(dimethylamino)-1-(7-(5-methylthiophen-2-yl)-2,7-diazaspiro[3.5]nonan- 2-yl)but-2-en-l-one (XS185-101). Example 95 was synthesized following similar procedure for preparing example 28. White solid, 18% yield.1H NMR (400 MHz, Methanol-64) 8 6.82 - 6.68 (m, 1H), 6.64 - 6.58 (m, 1H), 6.58 - 6.47 (m, 2H), 4.84 (s, 2H), 4.60 - 4.50 (m, 1H), 4.41 - 4.27 (m, 2H), 4.09 (d, J= 10.5 Hz, 1H), 4.01 - 3.94 (m, 2H), 3.93 - 3.84 (m, 2H), 2.93 (s, 6H), 2.64 - 2.57 (m, 2H), 2.43 (s, 3H). MS (ESI) [M+H]+=334.2.

[0550] Example 97: (£)-4-(dimethylamino)-l-(2-(5-methylthiophen-2-yl)-2,7-diazaspiro[3.5]nonan- 7-yl)but-2-en-l-one (XS185-113). Example 97 was synthesized following similar procedure for preparing example 53. White solid, 43% yield. ’H NMR (400 MHz, Methanol-6 / 4) 8 6.76 - 6.61 (m, 1H), 6.50 - 6.39 (m, 1H), 6.34 (s, 1H), 4.51 - 4.37 (m, 2H), 4.22 (t, J= 9.6 Hz, 1H), 4.18 - 4.08 (m, 1H), 3.96 - 3.83 (m, 2H), 3.61 - 3.49 (m, 1H), 2.86 (s, 6H), 2.60 (t, J= 5.9 Hz, 2H), 2.53 - 2.40 (m, 2H), 1.85 - 1.63 (m, 4H). MS (ESI) [M+H]+=348.2.

[0551] Example 98: (E)-2-(4-(dimethylamino)but-2-enoyl)-JV-(4,5,6,7-tetrahydrobenzo[ / »]thiophen- 2-yl)-2-azaspiro[3.3]heptane-6-carboxamide (XS185-114). Example 98 was synthesized following similar procedure for preparing example 53. White solid, 49% yield.JH NMR (400 MHz, Methanol-d4) 5 6.68 - 6.51 (m, 1H), 6.40 - 6.30 (m, 1H), 6.23 (s, 1H), 4.22 (d, J= 26.3 Hz, 2H), 3.96 (d, ,7 = 22.1 Hz, 2H), 3.89 - 3.79 (m, 2H), 3.12 - 2.94 (m, 1H), 2.79 (s, 6H), 2.54 (d, J = 5.7 Hz, 2H), 2.46 - 2.25 (m, 6H), 1.81 - 1.58 (m, 4H). MS (ESI) [M+H]+=388.2.

[0552] Example 99: (E)-4-(4-(dimethylamino)but-2-enoyl)-l-(lH-indol-2-yl)piperazin-2-one (XS185-116). Example 99 was synthesized following similar procedure for preparing example 28. White solid, 37% yield. ’H NMR (400 MHz, Methanol-d4) 8 7.47 - 7.36 (m, 1H), 7.32 (d, J = 8.0 Hz, 1H), 7.22 - 7.00 (m, 2H), 7.00 - 6.91 (m, 1H), 6.91 - 6.62 (m, 2H), 4.54 - 4.14 (m, 2H), 4.03 (q, J= 7.1, 6.2 Hz, 1H), 3.99 - 3.83 (m, 3H), 3.83 - 3.65 (m, 2H), 2.88 - 2.78 (m, 6H). MS (ESI) [M+H]+=327.2. 53. White solid, 21% yield. 'H NMR (400 MHz, Methanol-d)48 6.78 - 6.65 (m, 1H), 6.52 - 6.43 (m, 1H), 4.50 - 4.31 (m, 2H), 4.25 - 4.04 (m, 3H), 3.98 - 3.87 (m, 2H), 3.49 - 3.34 (m, 1H), 2.89 (s, 6H), 2.02 - 1.84 (m, 2H), 1.81 - 1.53 (m, 4H), 1.52 - 1.39 (m, 2H). MS (ESI) [M+H]+=280.2.

[0553] Example 103: (E)-JN,N-dimethyl-4-(4-(5-methylthiophen-2-yl)-3-oxopiperazin-l-yl)but-2- enamide (XS185-134). Example 103 was synthesized following similar procedure for preparing example 90. White solid, 19% yield. ’H NMR (400 MHz, Methanol-d4) 8 6.98 (d, J = 15.0 Hz, 1H), 6.74 - 6.59 (m, 3H), 4.16 - 4.02 (m, 6H), 3.72 (t, J= 5.7 Hz, 2H), 3.17 (s, 3H), 3.03 (s, 3H), 2.41 (s, 3H). MS (ESI) [M+H]1=308.2.

[0554] Scheme 11. Synthesis of example 104

[0555] Example 104: (2-(4-(5-methylthiophen-2-yl)-3-oxopiperazin-l-yl)-2-oxoethyl methylsulfamate (XS185-135). To a solution of intermediate 1 (20 mg, 0.102 mmol) dissolved in DMF (1 mL), and the HATU( 42.7 mg, 0.112 mmol, 1.1 eq), DIPEA(0.1 mL), 2-hydroxyacetic acid(9.4 mg, 0.102 mmol, 1.0 eq) were added. The reaction mixture stirred at rt 1 h, the crude mixtures were purified by HPLC yield the intermediate 10.

[0556] Then the intermediate 10 was re-dissovled in DCM (1 mL), EtsN (0.1 mL, 2.0 eq) and methyl sulfamoyl chloride (9.3 mg , 1.2 eq) were added. The reaction mixture stirred at rt for 5 h. Resulting crude mixtures were purified by HPLC yield the title example 104. White solid, 29% yield. ’H NMR (400 MHz, Methanol-d4) 8 6.75 - 6.51 (m, 2H), 4.91 (s, 2H), 4.38 (s, 2H), 3.95 (m, 4H), 2.75 (s, 3H), 2.41 (s, 3H). MS (ESI) [M+H]+=348.1.

[0557] Example 105: (E)-4-(4-(5-methylthiophen-2-yl)-3-oxopiperazin-l-yl)but-2-enenitrile

[0558] (XS185-138). Example 105 was synthesized following similar procedure for preparing example 90. White solid, 30% yield.XH NMR (400 MHz, Methanol^) 8 6.84 - 6.61 (m, 1H), 6.57 (d, J = 3.9 Hz, 1H), 6.55 - 6.48 (m, 1H), 5.99 - 5.77 (m, 1H), 3.93 - 3.86 (m, 2H), 3.80 - 3.72 (m, 1H), 3.66 (d, J= 6.5 Hz, 2H), 3.64 - 3.59 (m, 1H), 3.34 - 3.24 (m, 2H), 2.34 (s, 3H). MS (ESI) [M+Hf =262.1.

[0559] Example 106: l-(5-methylthiophen-2-yl)-4-(2-(phenylsulfonyl)acetyl)piperazin-2-one (XS185-140). Example 106 was synthesized following similar procedure for preparing example 104. White solid, 35% yield. 'H NMR (400 MHz, Methanol-d4) 8 8.03 - 7.90 (m, 2H), 7.82 - 7.53 (m, 3H), 6.68 - 6.51 (m, 2H), 4.58 (d, J= 18.3 Hz, 2H), 4.36 (d, J= 22.2 Hz, 2H), 4.06 (t, J = 5.4 Hz, 1H), 4.00 - 3.89 (m, 2H), 3.78 (t, J = 5.5 Hz, 1H), 2.42 (s, 3H). MS (ESI) [M+H]+=380.1.

[0560] Example 107: (E)-l-(7-(benzo[Z>]thiophen-2-yl)-2,7-diazaspiro[3.5]nonan-2-yl)-4-

[0561] (dimethylamino)but-2-en-l-one (XS185-147). Example 107 was synthesized following similar procedure for preparing example 28. White solid, 41% yield. 'H NMR (400 MHz, Methanol-d4) 8 7.54 (d, J= 7.9 Hz, 1H), 7.41 (d, J= 7.9 Hz, 1H), 7.16 (t, J= 7.6 Hz, 1H), 7.07 - 6.95 (m, 1H), 6.79 - 6.63 (m, 1H), 6.55 - 6.40 (m, 1H), 4.06 (s, 2H), 3.94 - 3.89 (m, 2H), 3.81 (s, 2H), 3.26 - 3.16 (m, 4H), 2.88 (s, 7H), 1.93 (t, J= 5.6 Hz, 4H). MS (ESI) [M+H]+=370.2.

[0562] Example 108: (E)-l-(6-(benzo[Z»]thiophen-2-yl)-2,6-diazaspiro[3.4]octan-2-yl)-4-

[0563] (dimethylamino)but-2-en-l-one (XS185-149). Example 108 was synthesized following similar procedure for preparing example 28. White solid, 30% yield. ’H NMR (400 MHz, Methanol- d4) 8 7.51 (d, J = 7.9 Hz, 1H), 7.35 (d, J = 7.9 Hz, 1H), 7.13 (t, J= 7.6 Hz, 1H), 6.93 (t, J= 7.6 Hz, 1H), 6.79 - 6.37 (m, 3H), 4.35 - 4.21 (m, 2H), 4.15 - 3.96 (m, 2H), 3.96 - 3.85 (m, 2H), 3.58 - 3.47 (m, 2H), 3.40 (t, J= 7.0 Hz, 2H), 2.86 (s, 7H), 2.27 (t, J= 6.8 Hz, 2H).MS (ESI) [M+Hf =356.3.

[0564] Exam pie 109: (E)- l-(2-(benzo [b] thiophen-2-yl)-2,6-diazaspiro [3.4] octan-6-yl)-4-

[0565] (dimethylamino)but-2-en-l-one (XS185-150). Example 109 was synthesized following similar procedure for preparing example 28. White solid, 30% yield. *HNMR (400 MHz, Methanol-)d 847.56 (d, J = 8.0 Hz, 1H), 7.42 (d, J = 7.9 Hz, 1H), 7.23 - 7.14 (m, 1H), 7.03 (t, J= 7.6 Hz, 1H), 6.83 - 6.64 (m, 2H), 4.24 - 4.01 (m, 2H), 3.99 - 3.86 (m, 5H), 3.79 - 3.68 (m, 2H), 3.65 - 3.53 (m, 1H), 2.95 - 2.84 (m, 6H), 2.38 - 2.15 (m, 2H).MS (ESI) [M+H]+=356.3.

[0566] Example 110: (E)-7V-(l-(4-(dimethylamino)but-2-enoyl)azetidin-3-yl)-5-methylthiophene-2- carboxamide (XS185-171). Example 110 was synthesized following similar procedure for preparing example 104. White solid, 15% yield 'H NMR (400 MHz, D SO-d6) 8 8.54 - 8.35 (m, 1H), 7.34 (d, J= 4.1 Hz, 4H), 7.29 - 7.20 (m, 1H), 6.67 - 6.49 (m, 2H), 4.81 (d, J= 21.1 Hz, 2H),

[0567] 4.25 (s, 2H), 4.19 (d, J= 4.8 Hz, 2H), 3.91 - 3.82 (m, 2H), 3.82 - 3.69 (m, 2H), 2.36 (s, 3H).MS

[0568] (ESI) [M+H]+=424.2.

[0569] Example 111: (E')-A / -(l-(4-(dimethylamino)but-2-enoyl)azetidin-3-yl)-5-methylthiophene-2- carboxamide (XS190-9). Example 111 was synthesized following similar procedure for preparing example 79. White solid, 56% yield *H NMR (400 MHz, Methanol-d4) 8 7.47 (d, J = 3.7 Hz, 1H), 6.74 (d, J= 3.0 Hz, 1H), 6.71 - 6.57 (m, 1H), 6.44 (d, J= 15.3 Hz, 1H), 4.76 - 4.63 (m, 1H), 4.58 (t, J= 8.6 Hz, 1H), 4.36 - 4.28 (m, 1H), 4.27 - 4.18 (m, 1H), 4.07 - 3.96 (m, 1H), 3.87 (d, J= 7.1 Hz, 2H), 2.83 (s, 6H), 2.43 (s, 3H).MS (ESI) [M+H]+=307.4.

[0570] Example 112: (E)-\-( l-(4-(dimethylamin())biit-2-enoyl)azetidin-3-yl)-V.5- dimethylthiophene-2-carboxamide (XS190-27). Example 112 was synthesized following similar procedure for preparing example 79. White solid, 23% yieldXH NMR (400 MHz, Methanol-^) 8 7.54 (d, J = 3.7 Hz, 1H), 6.89 - 6.77 (m, 2H), 6.57 (d, J = 15.2 Hz, 1H), 4.81 - 4.70 (m, 1H), 4.67 (t, J = 8.5 Hz, 1H), 4.45 - 4.35 (m, 1H), 4.34 - 4.24 (m, 1H), 4.14 (d, J = 7.2 Hz, 3H), 3.16 (s, 9H), 2.51 (s, 3H).MS (ESI) [M+H]+=322.2.

[0571] Example 113: (E)-3-(benzo[b]thiopheii-2-yl)-8-(4-(dimethylamino)but-2-enoyl)-3,8- diazabicyclo[3.2.1]octan-2-one (XS190-38). Example 113 was synthesized following similar procedure for preparing example 28. White solid, 25% yield1H NMR (400 MHz, Methanol-d4)

[0572] 8 7.72 (d, J= 7.8 Hz, 1H), 7.65 (d, J= 7.6 Hz, 1H), 7.34 - 7.19 (m, 2H), 7.03 - 6.69 (m, 3H), 5.07 (t, J= 35.5 Hz, 2H), 4.12 (t, J= 1 1 .3 Hz, 1H), 3.96 (s, 2H), 3.89 - 3.71 (m, 1H), 2.89 (s, 6H), 2.51 - 2.16 (m, 3H), 2.16 - 1.95 (m, 1H).MS (ESI) [M+H]+=370.2.

[0573] Example 114: (E)-3-(benzo[ / >]thiophen-3-yl)-8-(4-(dimethylamino)but-2-enoyl)-3,8- diazabicyclo[3.2.1]octan-2-one (XS190-44). Example 114 was synthesized following similar procedure for preparing example 28.1H NMR (400 MHz, Methano)l- 8d47.84 (d, J= 4.2 Hz, 1H), 7.58 (s, 1H), 7.41 - 7.28 (m, 3H), 6.97 - 6.75 (m, 2H), 5.05 - 4.88 (m, 2H), 4.08 - 3.80 (m, 3H), 3.57 - 3.39 (m, 1H), 2.83 (s, 6H), 2.52 - 2.1 1 (m, 4H). White solid, 15% yield MS (ESI) [M+H]+=370.2.

[0574] Example 115: (£)-8-(4-(dimethylamino)but-2-enoyl)-3-(5-phenylthiophen-2-yl)-3,8- diazabicyclo[3.2.1]octan-2-one (XS190-45). Example 115 was synthesized following similar procedure for preparing example 28. White solid, 18% yieldJH NMR (400 MHz, Methanol -t) 8 7.49 - 7.41 (m, 2H), 7.23 (t, J= 7.7 Hz, 2H), 7.12 (t, J= 7.4 Hz, 1H), 7.05 (d, J= 4.1 Hz, 1H), 6.91 - 6.64 (m, 2H), 6.63 - 6.55 (m, 1H), 5.10 - 4.89 (m, 2H), 4.05 - 3.89 (m, 1H), 3.89 - 3.80 (m, 2H), 3.64 (dd, J= 25.2, 11.4 Hz, 1H), 2.80 (s, 3H), 2.78 (s, 3H), 2.38 - 2.02 (m, 3H), 2.04 - 1.81 (m, 1H).MS (ESI) [M+H]+=396.3.

[0575] Example 116: (£)-l-(3-(benzo[6]thiophene-2-carbonyl)-3,6-diazabicyclo[3.1.1]heptan-6-yl)- 4-(dimethylamino)but-2-en-l-one (XS190-46). Example 116 was synthesized following similar procedure for preparing example 79. White solid, 40% yield1H NMR (400 MHz, Methanol-d4) 6 7.79 (d, J= 7.4 Hz, 2H), 7.72 (s, 1H), 7.32 (p, J= 7.0 Hz, 2H), 6.74 - 6.57 (m, 1H), 6.56 - 6.36 (m, 1H), 4.69 (d, J= 17.5 Hz, 1H), 4.47 (d, J = 23.4 Hz, 1H), 4.13 (t, J = 8.2 Hz, 1H), 4.07 - 3.96 (m, 1H), 3.94 - 3.74 (m, 4H), 2.80 (s, 3H), 2.78 - 2.67 (m, 4H), 1.65 (d, J= 9.2 Hz, 1H). MS (ESI) [M+H]+=370.2.

[0576] Example 117: (E)-l-(3-(benzo[Z>]thiophene-3-carbonyl)-3,6-diazabicyclo[3.1.1]heptan-6-yl)- 4-(dimethylamino)but-2-en-l-one (XS190-47). Example 117 was synthesized following similar procedure for preparing example 79. White solid, 50% yield1H NMR (400 MHz, Methanol-d)4 8 7.91 - 7.81 (m, 1H), 7.75 (d, J= 6.4 Hz, 1H), 7.64 (d, J= 5.6 Hz, 1H), 7.38 - 7.26 (m, 2H), 6.75 - 6.57 (m, 1H), 6.59 - 6.28 (m, 1H), 4.73 (s, 1H), 4.50 (s, 1H), 4.31 - 4.06 (m, 1H), 4.01 - 3.81 (m, 3H), 3.78 - 3.61 (m, 2H), 3.54 (d, J= 11.8 Hz, 1H), 2.81 (s, 3H), 2.76 - 2.61 (m, 4H), 1.65 (d, J= 9.2 Hz, 1H).MS (ESI) [M+H]+=370.2.

[0577] Example 118: (E)-4-(dimethylamino)-l-(3-(4,5,6,7-tetrahydrobenzob]thiophene-2- carbonyl)-3,6-diazabicydo[3.1.1]heptan-6-yl)but-2-en-l-one (XS190-48). Example 118 was synthesized following similar procedure for preparing example 79. White solid, 46% yield1H NMR (400 MHz, Methanol-d4) 8 7. 15 (s, 1H), 6.70 - 6.56 (m, 1H), 6.46 (d, J= 15.2 Hz, 1H), 4.66 (s, 1H), 4.42 (s, 1H), 4.31 - 3.69 (m, 6H), 2.83 - 2.67 (m, 7H), 2.67 - 2.60 (m, 2H), 2.54 - 2.42 (m, 2H), 1.70 (dt, J= 10.4, 5.4 Hz, 4H), 1.58 (d, J= 9.2 Hz, 1H).MS (ESI) [M+H]+=374.2.

[0578] Example 119: (E)-4-(dimethylamino)-l-(3-(4,5,6,7-tetrahydrobenzo[b]thiophene-3- carbonyl)-3,6-diazabicyclo[3.1.1]heptan-6-yl)but-2-en-l-one (XS190-49). Example 119 was synthesized following similar procedure for preparing example 79. White solid, 39% yield1H NMR (400 MHz, Methanol-d4) δ 7.15 (d, J = 4.1 Hz, 1H), 6.69 - 6.54 (m, 1H), 6.52 - 6.30 (m, 1H), 4.59 (d, J= 51.2 Hz, 1H), 4.36 (d, J= 73.5 Hz, 1H), 3.88 - 3.73 (m, 4H), 3.67 - 3.52 (m, 1H), 3.45 (d, J= 12.0 Hz, 1H), 2.79 - 2.70 (m, 6H), 2.70 - 2.58 (m, 3H), 2.39 - 2.28 (m, 2H), 1.77 - 1.59 (m, 4H), 1.59 - 1.49 (m, 1H).MS (ESI) [M+H]+=374.2.

[0579] Example 120: (E)-l-(3-(5,6-dihydro-4 / / -cyclopenta[Z»]thiophene-2-carbonyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl)-4-(dimethylamino)but-2-en-l-one (XS190-50). Example 120 was synthesized following similar procedure for preparing example 79. White solid, 55% yield

[0580] NMR (400 MHz, Methanol-d4) 8 7.26 (s, 1H), 6.72 - 6.56 (m, 1H), 6.49 (d, J= 15.1 Hz, 1H), 4.70 (s, 1H), 4.46 (s, 1H), 4.36 - 3.72 (m, 6H), 2.86 - 2.69 (m, 9H), 2.69 - 2.57 (m, 2H), 2.41 - 2.24 (m, 2H), 1.62 (d, J= 9.2 Hz, 1H).MS (ESI) [M+H]+=360.3.

[0581] Example 121: (£)-JV-(2-(6-(4-(dimethylamino)but-2-enoyl)-3,6-diazabicyclo[3.1.1Jheptane- 3-carbonyl)benzo[Z»]thiophen-5-yl)acetamide (XS190-59). Example 121 was synthesized following similar procedure for preparing example 79. White solid, 26% yield1H NMR (400 MHz, Methanol-d4)1H NMR (400 MHz, Methanol-d)48 8.22 (s, 1H), 7.74 (d, J= 8.7 Hz, 1H), 7.69 (s, 1H), 7.39 (t, J= 9.9 Hz, 1H), 6.75 - 6.62 (m, 1H), 6.58 - 6.41 (m, 1H), 4.72 (d, J= 17.8 Hz, 1H), 4.50 (d, J = 23.8 Hz, 1H), 4.38 - 3.75 (m, 6H), 2.83 (s, 3H), 2.80 - 2.71 (m, 4H), 2.08 (s, 3H), 1.68 (d, J= 9.2 Hz, 1H).MS (ESI) [M+H]+=427.3.

[0582] Example 122: (E')-4-(dimethylamino)-l-(3-(5-(methylamino)benzo[B]thiophene-2- carbonyl)-3,6-diazabicyclo[3.1.1]heptan-6-yl)but-2-en-l-one (XS190-68). Example 122 was synthesized following similar procedure for preparing example 79. White solid, 33% yield 'H NMR (400 MHz, Methanol-d4) 8 8.06 - 7.98 (m, 1H), 7.91 (d, J = 7.4 Hz, 1H), 7.83 (d, J = 6.2 Hz, 1H), 7.42 (d, J= 8.6 Hz, 1H), 6.76 - 6.64 (m, 1H), 6.58 - 6.44 (m, 1H), 4.73 (d, J= 18.3 Hz, 1H), 4.50 (d, J= Tl. 1 Hz, 1H), 4.41 - 3.81 (m, 6H), 3.07 - 3.00 (m, 3H), 2.84 - 2.74 (m, 7H), 1.69 (d, J= 9.2 Hz, 1H).MS (ESI) [M+H]+=399.2.

[0583] Example 123: (£)-l-(3-(5-acetyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridine-2-carbonyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl)-4-(dimethylamino)but-2-en-l-one (XS190-69). Example 123 was synthesized following similar procedure for preparing example 79. White solid, 22% yield 'H NMR (400 MHz, Methanol-d)45 7.26 (d, J= 10.6 Hz, 1H), 6.67 - 6.53 (m, 1H), 6.46 (dd, J = 14.7, 6.7 Hz, 1H), 4.66 (s, 1H), 4.53 - 4.36 (m, 3H), 4.31 - 3.94 (m, 2H), 3.93 - 3.64 (m, 6H), 2.84 - 2.78 (m, 1H), 2.78 - 2.62 (m, 8H), 2.14 - 1.99 (m, 3H), 1.58 (d, J= 9.2 Hz, 1H).MS (ESI) [M+H]+=417.3.

[0584] Example 124: (E)-3-(5-acetyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yI)-8-(4-

[0585] (dimethylamino)but-2-enoyl)-3,8-diazabicyclo[3.2.1]octan-2-one (XS190-70). Example 124 was synthesized following similar procedure for preparing example 79. White solid, 43% yield ’H NMR (400 MHz, Methanol-d)4δ 6.94 - 6.63 (m, 2H), 6.47 - 6.32 (m, 1H), 5.10 - 4.94 (m, 1H), 4.46 (s, 2H), 4.03 - 3.83 (m, 3H), 3.81 - 3.75 (m, 1H), 3.75 - 3.69 (m, 1H), 3.69 - 3.53 (m, 2H),

[0586] 2.85 - 2.78 (m, 6H), 2.77 - 2.60 (m, 2H), 2.40 - 2.15 (m, 2H), 2.11 (s, 3H), 2.07 (s, 1H), 2.03 -

[0587] 1.86 (m, 1H).MS (ESI) [M+H]+=417.3. Example 125: (E)-l-(4-(dimethylamino)but-2-enoyl)-Wmethyl-W(5-methylthiophen-2- yl)azetidine-3-carboxamide (XS190-75). Example 125 was synthesized following similar procedure for preparing example 79. White solid, 26% yield 'H NMR (400 MHz, Methanol-tZr) 8 6.84 - 6.70 (m, 1H), 6.51 (d, J= 15.1 Hz, 1H), 6.44 (s, 2H), 4.52 - 4.39 (m, 2H), 4.23 (t, J = 9.7 Hz, 1H), 4.19 - 4.04 (m, 3H), 3.60 - 3.48 (m, 1H), 3.10 (s, 9H), 2.32 (s, 3H).MS (ESI) [M+H]+=322.4. (E)-N,N,N,-trimethyl-4-(3-(methyl(5-methylthiophen-2- yl)carbamoyl)azetidin-l-yl)-4-oxobut-2-en-l-aminium (XS190-75-2). Example 126was synthesized following similar procedure for preparing example 79. White solid, 15% yield!H NMR (400 MHz, Methanol-d4) 8 6.79 - 6.68 (m, 1H), 6.67 (d, ,7 = 3.6 Hz, 1H), 6.60 - 6.55 (m,

[0588] 1H), 6.43 (d, J= 15.1 Hz, 1H), 4.41 - 4.33 (m, 1H), 4.17 (t, J= 8.9 Hz, 1H), 4.11 - 4.02 (m, 3H), 3.88 (t, J = 9.7 Hz, 1H), 3.71 - 3.59 (m, 1H), 3.18 (s, 3H), 3.06 (s, 9H), 2.39 (s, 3H).MS (ESI) [M+H]+=322.4.

[0589] Example 127: (E)-4-(dimethylamino)-l-(5-(5-methylthiophene-2-carbonyl)-2,5- diazabicyclo[2.2.2]octan-2-yl)but-2-en-l-one (XS190-77). Example 127 was synthesized following similar procedure for preparing example 79. White solid, 46% yield1H NMR (400 MHz, Methanol-^) 8 7.24 (d, J= 75.4 Hz, 1H), 6.91 - 6.54 (m, 3H), 4.73 (d, J = 24.1 Hz, 1H), 4.62 - 4.33 (m, 1H), 4.11 - 3.71 (m, 4H), 3.69 - 3.49 (m, 2H), 2.81 (s, 6H), 2.41 (s, 3H), 2.16 - 1.70 (m,

[0590] Example 128: (£)-4-(dimethylamino)-l-((1S,4S)-5-(5-methylthiophene-2-carbonyl)-2,5- diazabicyclo[2.2.1]heptan-2-yl)but-2-en-l-one (XS190-80). Example 128 was synthesized following similar procedure for preparing example 79. White solid, 42% yield1H NMR (400 MHz, Methanol-^) 8 7.31 - 7.14 (m, 1H), 6.80 - 6.34 (m, 3H), 4.86 - 4.78 (m, 2H), 3.80 (h, J= 18.8, 15.5 Hz, 3H), 3.67 - 3.31 (m, 3H), 2.73 (d, J= 7.4 Hz, 6H), 2.34 (s, 3H), 2.03 - 1.77 (m, 2H).MS (ESI) [M+H]+=334.1.

[0591] Example 129: (E')-4-(dimethylamino)-l-((lR,4R)-5-(5-methylthiophene-2-carbonyl)-2,5- diazabicyclo[2.2.1]heptan-2-yl)but-2-en-l-one (XS190-81). Example 129 was synthesized following similar procedure for preparing example 79. White solid, 59% yield1H NMR (400 MHz, Methanol-^) 8 7.35 - 7.18 (m, 1H), 6.90 - 6.40 (m, 3H), 4.87 - 4.80 (m, 2H), 3.98 - 3.71 (m, 3H), 3.72 - 3.32 (m, 3H), 2.82 - 2.70 (m, 6H), 2.36 (s, 3H), 2.03 - 1.78 (m, 2H).MS (ESI) [M+H]- =334.1.

[0592] Example 131 (£)-JV-((l-(4-(dimethylamino)but-2-enoyl)azetidin-2-yl)methyl)-5- methylthiophene-2-carboxamide (XS190-127). Example 131 was synthesized following similar procedure for preparing example 79. White solid, 34% yield 'H NMR (400 MHz, Methanol-6 / 4) 8 7.55 - 7.42 (m, 1H), 6.85 - 6.63 (m, 2H), 6.59 - 6.41 (m, 1H), 4.78 - 4.57 (m, 1H), 4.24 (t, J = 7.8 Hz, 1H), 4.02 - 3.89 (m, 2H), 3.88 - 3.73 (m, 2H), 3.69 - 3.50 (m, 1H), 2.88 (d, J= 11.0 Hz, 6H), 2.59 - 2.41 (m, 4H), 2.24 - 2.08 (m, 1H).MS (ESI) [M+H]+=322.2.

[0593] Example 132 (E)-l-(3-(cyclopentanecarbonyl)-3,6-diazabicyclo[3.1.1]heptan-6-yl)-4- (dimethylamino)but-2-en-l-one (XS190-128). Example 132 was synthesized following similar procedure for preparing example 79. White solid, 34% yield1H NMR (400 MHz, Methanol-d4) δ 6.78 - 6.63 (m, 1H), 6.61 - 6.48 (m, 1H), 4.75 (s, 1H), 4.52 (s, 1H), 4.20 - 3.66 (m, 6H), 3.05 - 2.94 (m, 1H), 2.88 (d, J = 2.7 Hz, 6H), 2.83 - 2.74 (m, 1H), 1.95 - 1.80 (m, 2H), 1.80 - 1.55 (m, 7H).MS (ESI) [M+Hf =306.2.

[0594] Example 133 (E)-l-(3-(cyclohexanecarbonyl)-3,6-diazabicyclo[3.1.1]heptan-6-yl)-4- (dimethylamino)but-2-en-l-one (XS190-129). Example 133 was synthesized following similar procedure for preparing example 79. White solid, 45% yield 'H NMR (400 MHz, Methanol-d4) 8 6.84 - 6.66 (m, 1H), 6.62 - 6.46 (m, 1H), 4.75 (s, 1H), 4.52 (s, 1H), 4.24 - 3.59 (m, 6H), 2.89 (d, J = 5.2 Hz, 6H), 2.84 - 2.74 (m, 1H), 2.63 - 2.47 (m, 1H), 1.86 - 1.58 (m, 6H), 1.54 - 1.13 (m, 5H).MS (ESI) [M+Hf =320.2.

[0595] Example 134 (E)-.-N-((5-(6-(4-(dimethylamino)but-2-enoyl)-3,6-diazabicyclo[3.1.1]heptane-3- carbonyl)thiophen-2-yl)methyl)acetamide (XS190-130). Example 134 was synthesized following similar procedure for preparing example 79. White solid, 22% yield1H NMR (400 MHz, Methanol-^) 8 7.53 - 7.40 (m, 1H), 7.01 (d, J = 3.3 Hz, 1H), 6.84 - 6.70 (m, 1H), 6.59 (d, J = 15.1 Hz, 1H), 4.80 (s, 1H), 4.54 (s, 3H), 4.47 - 3.86 (m, 6H), 2.98 - 2.76 (m, 7H), 1.99 (s, 3H), 1.74 (d, J= 9.2 Hz, 1H).MS (ESI) [M+H]+=391.2.

[0596] Example 135 (E)-4-(dimethylamino)-l-(3-(5-methyloxazole-4-carbonyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl)but-2-en-l-one (XS190-137). Example 135 was synthesized following similar procedure for preparing example 79. White solid, 47% yield1H NMR (400 MHz, Methanol-^) 8 8.05 (d, J = 3.4 Hz, 1H), 6.85 - 6.73 (m, 1H), 6.67 - 6.54 (m, 1H), 4.77 (s, 1H), 4.58 (d, J= 31.8 Hz, 1H), 4.49 - 4.23 (m, 2H), 4.09 - 3.88 (m, 4H), 2.93 (d, J= 3.0 Hz, 6H), 2.85 (d, J= 5.8 Hz, 1H), 2.54 (s, 3H), 1.80 - 1.71 (m, 1H).MS (ESI) [M+H]+=319.2.

[0597] Example 136 (E)-4-(dimethylamino)-l-(3-(5-(methoxymethyl)thiophene-2-carbonyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl)but-2-en-l-one (XS190-138). Example 136 was synthesized following similar procedure for preparing example 79. White solid, 38% yield 'H NMR (400 MHz, Methanol-^) 8 7.49 (d, J= 3.8 Hz, 1H), 7.05 (d, J= 3.8 Hz, 1H), 6.83 - 6.71 (m, 1H), 6.59 (d, J = 15.2 Hz, 1H), 4.81 (s, 1H), 4.63 (s, 3H), 4.47 - 3.82 (m, 6H), 3.39 (s, 3H), 2.97 - 2.78 (m, 7H), 1.74 (d, J= 9.2 Hz, 1H).MS (ESI) [M+H]+=364.2.

[0598] Example 137 (E)-3-cyclopentyl-8-(4-(dimethylamino)but-2-enoyl)-3,8- diazabicyclo[3.2.1]octan-2-one (XS190-144). Example 137 was synthesized following similar procedure for preparing example 28. White solid, 19% yield 'H NMR (400 MHz, Methanol-d)48 6.81 - 6.44 (tn, 2H), 4.75 - 4.28 (m, 3H), 3.78 - 3.63 (m, 2H), 3.41 - 3.25 (m, 1H), 2.95 - 2.80 (m, 1H), 2.66 (s, 6H), 2.15 - 1.75 (m, 3H), 1.70 - 1.54 (m, 2H), 1.47 (s, 3H), 1.41 - 1.21 (m, 4H).MS (ESI) [M+Hf =306.2.

[0599] Example 138 (E)-4-(dimethylamino)-l-(3-(5-methylfuran-2-carbonyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl)but-2-en-l-one (XS190-157). Example 138 was synthesized following similar procedure for preparing example 104. White solid, 35% yield 'H NMR (400 MHz, Methanol-d4) δ 7.04 (d, J= 3.5 Hz, 1H), 6.78 - 6.65 (m, 1H), 6.58 (d, J= 15.3 Hz, 1H), 6.20 (s, 1H), 4.79 (s, 1H), 4.56 (s, 1H), 4.47 - 3.79 (m, 6H), 2.90 - 2.75 (m, 7H), 2.34 (s, 3H), 1.69 (d, J= 9.2 Hz, 1H).MS (ESI) [M+H]+=318.2.

[0600] Example 139 (E')-4-(dimethylamino)-l-(3-((5-methylthiophen-2-yl)methyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl)but-2-en-l-one (XS190-158). Example 139 was synthesized following similar procedure for preparing example 79. White solid, 23% yield1H NMR (400 MHz, Methanol-^) 8 7.14 (d, J = 3.5 Hz, 1H), 6.85 - 6.70 (m, 2H), 6.53 (d, J = 15.2 Hz, 1H), 4.82 - 4.53 (m, 4H), 3.96 (d, J= 1A Hz, 2H), 3.90 - 3.61 (m, 4H), 2.83 (s, 2H), 2.52 (s, 3H), 2.07 (d, J = 10.2 Hz, 1H).MS (ESI) [M+H]+=320.2.

[0601] Example 140 (E)-4-(dimethylamino)-l-(3-(3-methylcyclopentane-l-carbonyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl)but-2-en-l-one (XS190-170). Example 40 was synthesized following similar procedure for preparing example 79. White solid, 25% yield 'H NMR (400 MHz, Methanol-d4) δ 6.66 - 6.54 (m, 1H), 6.47 - 6.36 (m, 1H), 4.62 (s, 1H), 4.38 (s, 1H), 4.02 (d, J = 11.4 Hz, 1H), 3.86 - 3.69 (m, 4H), 3.57 (d, J= 10.1 Hz, 1H), 2.77 - 2.73 (m, 6H), 2.70 - 2.61 (m, 1H), 2.02 - 1.59 (m, 6H), 1.49 (d, J= 9.4 Hz, 1H), 1.24 - 1.03 (m, 2H), 0.92 - 0.86 (m, 3H).MS (ESI) [M+H]+=320.2.

[0602] Example 141 (E)-8-(4-(dimethyIamino)but-2-enoyl)-3-(6-methylpyridin-2-yl)-3,8- diazabicyclo[3.2.1]octan-2-one (XS190-181). Example 141 was synthesized following similar procedure for preparing example 28. White solid, 29% yield 'H NMR (400 MHz, Methanol-6 / 4) 5 7.84 - 7.60 (m, 1H), 7.46 - 7.07 (m, 1H), 6.99 - 6.72 (m, 2H), 6.72 - 6.47 (m, 1H), 4.97 (dd, J = 42.5, 7.7 Hz, 1H), 4.84 - 4.70 (m, 1H), 3.96 - 3.83 (m, 2H), 3.81 - 3.70 (m, 1H), 2.91 - 2.75 (m, 6H), 2.47 (d, J= 37.5 Hz, 3H), 2.39 - 2.15 (m, 3H), 2.14 - 1.79 (m, 1H).MS (ESI) [M+H]+=329.2.

[0603] Example 142 (E)-8-(4-(dimethylamino)but-2-enoyl)-3-(m-tolyl)-3,8- diazabicyclo[3.2.1]octan-2-one (XS190-182). Example 142 was synthesized following similar procedure for preparing example 28. White solid, 34% yield1H NMR (400 MHz, Methanol)-d 547.20 (t, J = 7.7 Hz, 1H), 7.05 (d, J = 7.5 Hz, 1H), 6.98 (s, 1H), 6.93 (d, J = 7.8 Hz, 1H), 6.90 - 6.66 (m, 2H), 4.98 - 4.88 (m, 1H), 4.83 - 4.70 (m, 1H), 3.95 - 3.75 (m, 3H), 3.39 (dd, J = 21.6, 11.8 Hz, 1H), 2.81 (s, 6H), 2.43 - 1.96 (m, 7H).MS (ESI) [M+H]+=328.2.

[0604] Example 143 (E')-8-(4-(dimethylamino)but-2-enoyl)-3-(5-methylthiazol-2-yl)-3,8- diazabicyclo[3.2.1]octan-2-one (XS190-183). Example 143 was synthesized following similar procedure for preparing example 28. White solid, 33% yield 'H NMR (400 MHz, Methanol-d)48 6.87 (s, 1H), 6.77 - 6.46 (m, 2H), 4.84 - 4.70 (m, 2H), 3.90 - 3.60 (m, 4H), 2.65 (s, 6H), 2.22 - 1.91 (m, 6H), 1.85 - 1.58 (m, 1H).MS (ESI) [M+H]+=335.2.

[0605] Example 146 (£)-8-(4-(dimethylamino)but-2-enoyl)-3-methyl-3,8-diazabicyclo[3.2.1]octan- 2-one (XS197-4). Example 146 was synthesized following similar procedure for preparing example 28. White solid, 13% yieldJH NMR (400 MHz, Methanol-d4) 8 6.88 - 6.62 (m, 2H), 4.82 - 4.49 (m, 2H), 3.84 (d, J= 7.4 Hz, 2H), 3.66 - 3.41 (m, 2H), 2.77 (d, J = 6.3 Hz, 6H), 2.72 (s, 3H), 2.28 - 1.70 (m, 4H).MS (ESI) [M+H]+=252.2.

[0606] Example 147 (E)-8-(4-(dimethylamino)but-2-enoyl)-3-methyl-3,8-diazabicyclo[3.2.1]octan- 2-one (XS197-5). Example 147 was synthesized following similar procedure for preparing example 79. White solid, 25% yield 'H NMR (400 MHz, Methanol-d4) 8 6.73 - 6.56 (m, 1H), 6.54 - 6.35 (m, 1H), 4.71 - 4.51 (m, 2H), 4.42 (s, 1H), 4.12 - 3.71 (m, 4H), 3.70 - 3.38 (m, 4H),

[0607] 2.83 - 2.73 (m, 6H), 2.73 - 2.63 (m, 1H), 2.25 - 2.08 (m, 1H), 2.05 - 1.79 (m, 5H), 1.78 - 1.46 (m, 2H).MS (ESI) [M+H]+=349.2.

[0608] Example 148 (E)-l-(3-(l-acetylpiperidine-4-carbonyl)-3,6-diazabicyclo[3.1.1]heptan-6-yl)-4- (dimethylamino)but-2-en-l-one (XS197-6). Example 148 was synthesized following similar procedure for preparing example 79. White solid, 23% yield 'H NMR (400 MHz, Methanol-t / 4) 6 6.68 - 6.53 (m, 1H), 6.47 - 6.35 (m, 1H), 4.62 (d, J= 12.9 Hz, 1H), 4.38 (s, 2H), 4.11 - 3.63 (m, 6H), 3.63 - 3.51 (m, 3H), 2.74 (d, J= 6.9 Hz, 6H), 2.68 - 2.48 (m, 2H), 2.01 (d, J= 48.1 Hz, 3H), 1.73 - 1.28 (m, 5H).MS (ESI) [M+H]+=363.2.

[0609] Example 149 (E)-8-(4-(dimethylamino)but-2-enoyl)-3-(5-methyl-lH-pyrrol-2-yl)-3,8- diazabicyclo[3.2.1]octan-2-one (XS197-14). Example 149 was synthesized following similar procedure for preparing example 28. White solid, 34% yield 'H NMR (400 MHz, Methanol-d)45 7.55 - 6.56 (m, 2H), 6.49 (d, J= 15.2 Hz, 2H), 5.83 (s, 1H), 4.71 (s, 1H), 4.48 (s, 1H), 3.83 (d, J = 6.5 Hz, 4H), 2.84 - 2.68 (m, 8H), 2.15 (s, 3H), 1.59 (d, .1= 9.1 Hz, 1H), 1.32 - 1.16 (m, 1H).MS (ESI) [M+H]+=317.2.

[0610] Example 150 (XS197-29). 8-(2-chloroacetyl)-3-methyl-3,8-diazabicyclo[3.2.1]octan-2-one

[0611] Example 150 was synthesized following similar procedure for preparing example 18. White solid, 33% yield1H NMR (400 MHz, Methanol-d4) 8 4.30 - 4.06 (m, 2H), 3.75 - 3.52 (m, 2H), 3.20 - 3.00 (m, 2H), 2.80 (s, 3H), 2.34 - 1.76 (m, 4H).MS (ESI) [M+H]+=217.1.

[0612] Example 151 (E)-l-(3-(cyclobutanecarbonyl)-3,6-diazabicyclo[3.1.1]heptan-6-yl)-4- (dimethylamino)but-2-en-l-one (XS197-38). Example 151 was synthesized following similar procedure for preparing example 79. White solid, 23% yield1H NMR (400 MHz, Methanol)-d 846.55 (dt, J= 14.7, 7.1 Hz, 1H), 6.41 (d, J= 15.2 Hz, 1H), 4.64 - 4.52 (m, 1H), 4.40 - 4.27 (m, 1H), 3.90 - 3.72 (m, 3H), 3.67 - 3.44 (m, 3H), 3.32 - 3.22 (m, 1H), 2.72 (s, 6H), 2.61 (t, J = 7.6 Hz, 1H), 2.14 - 1.96 (m, 4H), 1.91 - 1.81 (m, 1H), 1.73 - 1.57 (m, 1H), 1.46 (d, J= 9.2 Hz, 1H).MS (ESI) [M+H]+=292.2.

[0613] Example 152 (E)-4-(dimethylamino)-l-(3-(5-isopropylthiophene-2-carbonyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl)but-2-en-l-one (XS197-49). Example 152 was synthesized following similar procedure for preparing example 79. White solid, 46% yield 'H NMR (400 MHz, Methanol-d4) 8 7.48 (d, J= 3.9 Hz, 1H), 6.91 (d, ,7 = 3.9 Hz, 1H), 6.85 - 6.72 (m, 1H), 6.62 (d, J= 15.3 Hz, 1H), 4.82 (s, 1H), 4.65 - 4.51 (m, 1H), 4.48 - 3.91 (m, 6H), 3.22 (p, J= 6.9 Hz, 1H), 2.98 - 2.78 (m, 7H), 1.74 (d, J = 9.2 Hz, 1H), 1.35 (d, J = 6.9 Hz, 6H).MS (ESI) [M+Hf =362.2.

[0614] Example 153 (£')-4-(dimethylamino)-l-(3-isobutyryl-3,6-diazabicyclo[3.1.1]heptan-6-yl)but- 2-en-l-one (XS197-50). Example 153 was synthesized following similar procedure for preparing example 79. White solid, 27% yield 'H NMR (400 MHz, Methanol-d4) 8 6.71 - 6.58 (m, 1H), 6.55 - 6.41 (m, 1H), 4.68 (s, 1H), 4.44 (s, 1H), 3.91 - 3.81 (m, 3H), 3.70 - 3.56 (m, 3H), 3.14 (q, 7.4 Hz, 1H), 2.81 (s, 6H), 2.74 - 2.68 (m, 1H), 1.59 - 1.53 (m, 1H), 1.08 - 0.96 (m, 6H).MS (ESI) [M+H]+=280.2.

[0615] Example 154 (£)-3-cyclobutyl-8-(4-(dimethylamino)but-2-enoyl)-3,8- diazabicyclo[3.2.1]octan-2-one (XS197-70). Example 154 was synthesized following similar procedure for preparing example 79. White solid, 21% yield 'H NMR (400 MHz, Methanol-d4) 6

[0616] 6.83 - 6.55 (m, 2H), 4.81 - 4.43 (m, 3H), 3.88 - 3.70 (m, 2H), 3.52 - 3.37 (m, 1H), 3.15 - 3.07

[0617] (m, 1H), 2.75 (d, J= 6.0 Hz, 6H), 2.25 - 1.83 (m, 7H), 1.83 - 1.65 (m, 1H), 1.62 - 1.46 (m, 2H).MS

[0618] (ESI) [M+H]+=292.2.

[0619] Example 155 (£)-l-(3-(5-(l-acetylpyrrolidin-3-yl)thiophene-2-carbonyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl)-4-(dimethylamino)but-2-en-l-one (XS197-71). Example 155 was synthesized following similar procedure for preparing example 79. White solid, 26% yield ’H NMR (400 MHz, Methanol-d4) 8 7.41 (s, 1H), 6.98 - 6.88 (m, 1H), 6.74 - 6.61 (m, 1H), 6.55 - 6.42 (m, 1H), 4.72 (s, 1H), 4.47 (s, 1H), 4.11 - 3.79 (m, 6H), 3.71 - 3.52 (m, 3H), 3.40 - 3.29 (m, 1H), 3.14 (q, J = 7.4 Hz, 1H), 2.81 (s, 6H), 2.78 - 2.74 (m, 1H), 2.43 - 2.25 (m, 1H), 2.13 - 1.93 (m, 4H), 1.65 (d, J = 9.2 Hz, 1H).MS (ESI) [M+H]+=431.2.

[0620] Example 156 (£)-l-(3-(5-(l-acetylpiperidin-4-yl)thiophene-2-carbonyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl)-4-(dimethylamino)but-2-en-l-one (XS197-72). Example 156 was synthesized following similar procedure for preparing example 79. White solid, 25% yield 1 HMR400 MHz, Methanol-d4) 5 7.37 (d, J= 3.9 Hz, 1H), 6.83 (d, J= 4.0 Hz, 1H), 6.70 - 6.58 (m, 1H), 6.48 (d, J = 15.3 Hz, 1H), 4.69 (s, 1H), 4.47 (t, J = 14.4 Hz, 2H), 4.35 - 4.07 (m, 1H), 4.02 - 3.73 (m, 6H), 3.18 - 3.00 (m, 2H), 2.78 (s, 6H), 2.73 - 2.59 (m, 2H), 2.05 - 1.90 (m, 5H), 1.64 - 1.38 (m, 3H).MS (ESI) [M+H]1=445.2.

[0621] Example 157 (£)-4-(dimethylamino)-l-(3-(5-(l-methylpyrrolidin-3-yl)thiophene-2- carbonyl)-3,6-diazabicyclo[3.1.1]heptan-6-yl)but-2-en-l-one (XS197-73). Example 157 was synthesized following similar procedure for preparing example 79. White solid, 20% yield 'H NMR (400 MHz, Methanol-6 / 4) 5 7.46 (d, J= 4.3 Hz, 1H), 7.02 (s, 1H), 6.77 - 6.64 (m, 1H), 6.53 (d, J= 15.0 Hz, 1H), 4.74 (s, 1H), 4.51 (d, J= 19.5 Hz, 1H), 4.39 - 4.12 (m, 1H), 4.09 - 3.95 (m, 2H), 3.93 - 3.72 (m, 5H), 3.68 - 3.49 (m, 1H), 3.21 - 3.08 (m, 1H), 2.95 (s, 3H), 2.83 (s, 6H), 2.81

[0622] - 2.70 (m, 2H), 2.67 - 2.45 (m, 1H), 2.35 - 2.07 (m, 1H), 1.67 (d, J = 9.2 Hz, 1H).MS (ESI) [M+H]+=403.2.

[0623] Example 158 (£)-4-(dimethylamino)-l-(3-(5-(l-methylpiperidin-4-yl)thiophene-2-carbonyl)- 3,6-diazabicyclo[3.1.1]heptan-6-yl)but-2-en-l-one (XS197-74). Example 158 was synthesized following similar procedure for preparing example 79. White solid, 17% yield 'H NMR (400 MHz, Methanol-d4) 8 7.43 (d, J= 4.1 Hz, 1H), 6.91 (d, J= 3.9 Hz, 1H), 6.76 - 6.62 (m, 1H), 6.51

[0624] (d, J= 15.1 Hz, 1H), 4.72 (s, 1H), 4.49 (d, J= 18.1 Hz, 1H), 4.32 - 4.07 (m, 1H), 4.01 (d, J= 11.4

[0625] Hz, 1H), 3.88 - 3.82 (m, 3H), 3.58 - 3.47 (m, 2H), 3.20 - 3.02 (m, 3H), 2.85 - 2.80 (m, 8H), 2.76

[0626] - 2.70 (m, 3H), 2.21 (d, J= 14.4 Hz, 2H), 1.96 - 1.78 (m, 2H), 1.65 (d, .7 = 9.2 Hz, 1H).MS (ESI) [M+H]1=417.2.

[0627] Example 159 (£)-4-(dimethylamino)-l-(3-(l-methylazetidine-3-carbonyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl)but-2-en-l-one (XS197-85). Example 159 was synthesized following similar procedure for preparing example 79. White solid, 19% yield 'H NMR (400

[0628] MHz, Methanol-d4) δ 6.70 - 6.56 (m, 1H), 6.48 - 6.36 (m, 1H), 4.70 - 4.61 (m, 1H), 4.47 - 4.32

[0629] (m, 3H), 4.11 - 3.98 (m, 2H), 3.94 - 3.79 (m, 4H), 3.75 - 3.65 (m, 2H), 2.86 - 2.79 (m, 4H), 2.79

[0630] - 2.75 (m, 6H), 2.73 - 2.64 (m, 1H), 1.57 (d, J= 9.2 Hz, 1H).MS (ESI) [M+H]+=307.2. Example 160 (E')-4-(dimethylamino)-l-(3-(5-(3-methoxyprop-l-yn-l-yl)thiophene-2- carbonyl)-3,6-diazabicyclo[3.1.1]heptan-6-yl)but-2-en-l-one (XS197-93). Example 160 was synthesized following similar procedure for preparing example 79. White solid, 44% yield!H NMR (400 MHz, Methanol-d4) 8 7.39 (d, J = 4.0 Hz, 1H), 7.12 (d, J = 4.1 Hz, 1H), 6.69 - 6.58 (m, 1H), 6.53 - 6.39 (m, 1H), 4.69 (s, 1H), 4.46 (d, J= 16.4 Hz, 1H), 4.34 - 4.18 (m, 3H), 4.10 - 3.92 (m, 2H), 3.87 - 3.79 (m, 3H), 3.29 (s, 3H), 2.84 - 2.65 (m, 7H), 1.62 (d, J= 9.3 Hz, 1H).MS (ESI) [M+H]+=388.2.

[0631] Example 161 (£)-4-(dimethylamino)-l-(3-(5-(3-hydroxyprop-l-yn-l-yl)thiophene-2- carbonyl)-3,6-diazabicyclo[3.1.1]heptan-6-yl)but-2-en-l-one (XS197-94). Example 161 was synthesized following similar procedure for preparing example 79. White solid, 28% yield 'H NMR (400 MHz, Methanol-d4) δ 7.24 (d, J = 3.9 Hz, 1H), 6.94 (d, J = 4.0 Hz, 1H), 6.58 - 6.44 (m, 1H), 6.37 - 6.23 (m, 1H), 4.54 (s, 1H), 4.31 (d, J= 14.2 Hz, 1H), 4.20 - 4.07 (m, 2H), 3.97 - 3.77 (m, 2H), 3.77 - 3.58 (m, 4H), 2.67 - 2.52 (m, 7H), 1.47 (d, J= 9.2 Hz, 1H).MS (ESI) [M+Hf =374.1.

[0632] Example 162 (£)-3-(5-(6-(4-(dimethylamino)but-2-enoyl)-3,6-diazabicyclo[3.1.1]heptane-3- carbonyl)thiophen-2-yl)prop-2-yn-l-yl acetate (XS197-96). Example 162 was synthesized following similar procedure for preparing example 79. White solid, 29% yield1H NMR (400 MHz, Methanol-^) 8 7.45 (d, J= 4.7 Hz, 1H), 7.19 (d, J= 3.9 Hz, 1H), 6.75 - 6.64 (m, 1H), 6.51 (t, J = 14.1 Hz, 1H), 4.92 - 4.88 (m, 1H), 4.73 (s, 1H), 4.51 (d, J= 17.4 Hz, 1H), 4.33 (d, J= 11.9 Hz, 1H), 4.19 - 3.98 (m, 2H), 3.96 - 3.80 (m, 4H), 2.90 - 2.71 (tn, 7H), 2.03 (s, 3H), 1.67 (d, J= 9.2 Hz, 1H).MS (ESI) [M+H]+=416.2.

[0633] Example 163 (E)-l-(3-(2-acetyl-2-azaspiro[3.3]heptane-6-carbonyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl)-4-(dimethylamino)but-2-en-l-one (XS197-133). Example 163 was synthesized following similar procedure for preparing example 79. White solid, 22% yield ’H NMR (400 MHz, Methanol-d4) δ 6.73 - 6.61 (m, 1H), 6.48 (d, J= 15.3 Hz, 1H), 4.67 (s, 1H), 4.46 (d, J= 11.9 Hz, 1H), 4. 18 (s, 1H), 4.05 (d, J= 4.1 Hz, 1H), 3.99 - 3.93 (m, 2H), 3.87 (d, J - 1.2 Hz, 2H), 3.80 (d, J= 8.3 Hz, 1H), 3.76 - 3.57 (m, 3H), 2.83 (d, J = 3.3 Hz, 6H), 2.78 - 2.68 (m, 2H), 2.45 - 2.27 (m, 4H), 1.77 (d, J= 6.7 Hz, 3H), 1.61 - 1.54 (m, 1H).MS (ESI) [M+H]+=375.2.

[0634] Example 164 (E)-3-(5-(l-acetylpiperidin-4-yl)thiophen-2-yl)-8-(4-(dimethylamino)but-2- enoyl)-3,8-diazabicyclo[3.2.1]octan-2-one (XS197-145). Example 164 was synthesized following similar procedure for preparing example 28. White solid, 25% yield1H NMR (400 MHz, Methanol-dr) 6 6.95 - 6.65 (m, 2H), 6.61 (d, J = 3.9 Hz, 1H), 6.57 - 6.46 (m, 1H), 5.01 (d, J = 38.0 Hz, 2H), 4.51 (d, J = 13.2 Hz, 1H), 4.06 - 3.83 (m, 4H), 3.64 (dd, J = 20.1, 11.7 Hz, 1H), 3.23 - 3.10 (m, 1H), 2.97 (t, J= 13.4 Hz, 1H), 2.84 (d, J= 5.2 Hz, 6H), 2.74 - 2.61 (m, 1H), 2.40 - 2.10 (m, 3H), 2.06 (s, 3H), 2.03 - 1.85 (m, 3H), 1.70 - 1.41 (m, 2H).MS (ESI) [M+H]+=445.2.

[0635]

[0636] Example 166 8-(4-(dimethylamino)but-2-ynoyl)-3-(5-methylthiophen-2-yl)-3,8- diazabicyclo[3.2.1]octan-2-one (XS197-185). Example 166 was synthesized following similar procedure for preparing example 28. White solid, 39% yield ’H NMR (400 MHz, Methanol-d)4δ 6.59 (d, J= 2.0 Hz, 2H), 5.12 - 4.91 (m, 2H), 4.41 - 4.34 (m, 2H), 4.14 - 4.00 (m, 1H), 3.81 - 3.65 (m, 1H), 3.11 - 3.01 (m, 6H), 2.41 (s, 3H), 2.40 - 2.32 (m, 1H), 2.31 - 2.16 (m, 2H), 2.12 - 2.00 (m, 1H).MS (ESI) [M+H]+=332.1.

[0637] Example 167 (£)-4-(dimethylamino)-l-(isoindolin-2-yl)but-2-en-l-one (XS197-186).

[0638] Example 167 was synthesized following similar procedure for preparing example 79. White solid , 60% yieldrH NMR (400 MHz, Methanol-d4) 8 7.33 - 7.20 (m, 4H), 6.86 - 6.67 (m, 2H), 4.94 (s, 2H), 4.75 (s, 2H), 3.94 (d, J= 6.2 Hz, 2H), 2.87 (s, 6H).MS (ESI) [M+H]+=231.2.

[0639] Example 168 (£)-l-(4-(dimethylamino)but-2-enoyl)-3-methyl-W-(5-methylthiophen-2- yl)azetidine-3-carboxamide (XS209-21). Example 168 was synthesized following similar procedure for preparing example 53. White solid, 43% yield1H NMR (400 MHz, Methanol)-d 846.77 - 6.65 (m, 1H), 6.55 (d, J= 3.5 Hz, 1H), 6.52 - 6.42 (m, 2H), 4.65 (d, J= 8.9 Hz, 1H), 4.35 (d, J= 10.6 Hz, 1H), 4.12 (d, J= 9.0 Hz, 1H), 3.99 - 3.85 (m, 3H), 2.89 (s, 6H), 2.37 (s, 3H), 1.65 (s, 3H).MS (ESI) [M+H]+=322.2.

[0640] Example 169 (E')-l-(4-(dimethylamino)but-2-enoyl)-3-hydroxy-AN-(5-methylthiophen-2- yl)azetidine-3-carboxamide (XS209-22). Example 169 was synthesized following similar procedure for preparing example 53. White solid, 29% yield 'H NMR (400 MHz, Methanol-d)48 6.84 - 6.71 (m, 1H), 6.68 (d, J = 3.6 Hz, 1H), 6.55 (d, J= 15.2 Hz, 2H), 4.71 (d, J= 9.4 Hz, 1H), 4.46 (d, J = 10.9 Hz, 1H), 4.29 (d, J= 9.4 Hz, 1H), 4.05 (d, J= 10.9 Hz, 1H), 3.97 (d, J= 7.1 Hz, 2H), 2.92 (s, 6H), 2.41 (s, 3H).MS (ESI) [M+H]+=324.1.

[0641] Example 170 ( / :)-l-(4-(dimetliylamiiio)biit-2-enioyl)-3-methyl-A-(5-methyloxazol-2- yl)azetidine-3-carboxamide (XS209-23). Example 170 was synthesized following similar procedure for preparing example 53. White solid, 38% yield ’H NMR (400 MHz, Methanol-d4) 8 6.81 (s, 1H), 6.79 - 6.66 (m, 1H), 6.50 (d, J= 15.2 Hz, 1H), 4.71 (d, J= 9.0 Hz, 1H), 4.39 (d, J = 10.6 Hz, 1H), 4.14 (d, J = 9.1 Hz, 1H), 4.00 - 3.89 (m, 3H), 2.91 (s, 6H), 2.32 (s, 3H), 1.68 (s, 3H).MS (ESI) [M+Hf =307.2. Example 174 (E)-4-(dimethylamino)-2V-(l-(5-methylthiophen-2-yl)-2-oxopiperidin-4-yl)but- 2-enamide (XS209-27). Example 174 was synthesized following similar procedure for preparing example 28. White solid, 27% yield 'H NMR (400 MHz, Methanol-d4) 8 6.81 - 6.66 (m, 1H), 6.63 - 6.51 (m, 2H), 6.37 (d, J = 15.3 Hz, 1H), 4.37 - 4.24 (m, 1H), 3.96 - 3.76 (m, 4H), 2.89 (s, 7H), 2.58 - 2.46 (m, 1H), 2.38 (s, 3H), 2.32 - 2.21 (m, 1H), 2.11 - 2.00 (m, 1H).MS (ESI) [M+Hf =322.2.

[0642] Example 175 (£)-4-(dimethylamino)-l-(3-propionyl-3,6-diazabicyclo[3.1.1]heptan-6-yl)but- 2-en-l-one (XS209-28). Example 175 was synthesized following similar procedure for preparing example 79. White solid, 21% yield *HNMR (400 MHz, Methanol-)d 846.76 - 6.62 (m, 1H), 6.53 (d, J= 15.2 Hz, 1H), 4.76 - 4.69 (m, 1H), 4.54 - 4.43 (m, 1H), 4.10 - 3.76 (m, 4H), 3.72 (d, J = 5.4 Hz, 1H), 3.66 (d, J= 6.9 Hz, 1H), 2.86 (s, 6H), 2.80 - 2.70 (m, 1H), 2.46 - 2.23 (m, 2H), 1.61 (d, J= 9.1 Hz, 1H), 1.06 (t, J= 7.4 Hz, 3H).MS (ESI) [M+H]+=266.2.

[0643] Example 176 (£)-3-(cyclohex-l-en-l-yl)-8-(4-(dimethylamino)but-2-enoyl)-3,8- diazabicyclo[3.2.1]octan-2-one (XS209-30). Example 176 was synthesized following similar procedure for preparing example 28. White solid, 11% yield 'H NMR (400 MHz, Methanol-d4) 8 6.95 - 6.71 (m, 2H), 5.64 (s, 1H), 4.97 - 4.88 (m, 1H), 4.82 - 4.64 (m, 1H), 4.00 - 3.90 (m, 2H), 3.80 - 3.62 (m, 1H), 3.27 - 3.06 (m, 1H), 2.90 (s, 6H), 2.42 - 2.04 (m, 6H), 2.03 - 1.92 (m, 1H), 1.75 - 1.64 (m, 2H), 1.64 - 1.52 (m, 2H), 1.52 - 1.32 (m, 1H).MS (ESI) [M+H]+=318.2. Example 177 (E)-l-(4-(dimethylamino)but-2-enoyl)-3-methyl-N-(5-methylthiazol-2- yl)azetidine-3-carboxamide (XS209-38). Example 177 was synthesized following similar procedure for preparing example 53. White solid, 44% yield1H NMR (400 MHz, M Methanol)-d 847.15 (s, 1H), 6.72 - 6.56 (m, 1H), 6.43 (d, J= 15.3 Hz, 1H), 4.65 (d, J= 9.1 Hz, 1H), 4.35 (d, J = 10.7 Hz, 1H), 4.12 (d, J= 9.2 Hz, 1H), 3.87 (d, J= 8.2 Hz, 3H), 2.83 (s, 6H), 2.35 (s, 3H), 1.62 (s, 3H).MS (ESI) [M+H]- =323.1.

[0644] Example 178 (E)-l-(4-(dimethylamino)but-2-enoyl)-3-hydroxy-JV-(5-methylthiazol-2- yl)azetidine-3-carboxamide (XS209-39). Example 178 was synthesized following similar procedure for preparing example 53. White solid, 27% yield1H NMR (400 MHz, Methanol)-d 847.13 (s, 1H), 6.81 - 6.66 (m, 1H), 6.49 (d, J = 15.3 Hz, 1H), 4.68 (d, J= 9.5 Hz, 1H), 4.43 (d, J = 11.0 Hz, 1H), 4.27 (d, J = 9.5 Hz, 1H), 4.03 (d, J= 11.0 Hz, 1H), 3.91 (d, J= 7.1 Hz, 2H), 2.87 (s, 6H), 2.37 (s, 3H).MS (ESI) [M+H]+=325.1.

[0645] Example 179 (E)-l-(3-butyryl-3,6-diazabicyclo[3.1.1]heptan-6-yl)-4-(dimethylamino)but-2- en-l-one (XS209-40). Example 179 was synthesized following similar procedure for preparing example 79. White solid, 19% yield1HNMR (400 MHz, Methanol-)d 846.84 - 6.69 (m, 1H), 6.57 (d, J = 15.3 Hz, 1H), 4.77 (s, 1H), 4.54 (s, 1H), 4.13 (d, J = 11.5 Hz, 1H), 4.02 - 3.87 (m, 3H), 3.81 - 3.69 (m, 2H), 2.91 (d, J= 4.4 Hz, 6H), 2.82 (q, J= 6.6 Hz, 1H), 2.47 - 2.23 (m, 2H), 1.72 - 1.58 (m, 3H), 0.98 (t, J= 13 Hz, 3H).MS (ESI) [M+H]+=280.2.

[0646] Example 180 (£)-l-(3-(2-acetyl-2-azaspiro[4.5]decane-8-carbonyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl)-4-(dimethylamino)but-2-en-l-one (XS209-53). Example 180 was synthesized following similar procedure for preparing example 79. White solid, 18% yield 'H NMR (400 MHz, Methanol-d4) 86.67 - 6.54 (m, 1H), 6.47 (d, J= 15.2 Hz, 1H), 4.66 (s, 1H), 4.42 (s, 1H), 4.12 - 3.91 (m, 1H), 3.87 - 3.77 (m, 3H), 3.68 - 3.51 (m, 4H), 3.51 - 3.40 (m, 1H), 3.15 (d, J = 5.7 Hz, 1H), 2.78 (s, 6H), 2.74 (s, 3H), 2.71 - 2.65 (m, 1H), 2.59 - 2.46 (m, 1H), 2.13 - 2.02 (m, 3H), 1.90 - 1.81 (m, 1H), 1.63 - 1.50 (m, 5H), 1.46 - 1.37 (m, 2H).MS (ESI) [M+H]+

[0647] =417.3.

[0648] Example 181 (E)-l-(3-(2-acetyl-2-azaspiro[3.5]nonane-7-carbonyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl)-4-(dimethylamino)but-2-en-l-one (XS209-54). Example 181 was synthesized following similar procedure for preparing example 79. White solid, 21% yield!H NMR (400 MHz, Methanol-d4) δ 6.71 - 6.60 (m, 1H), 6.51 (d, J= 15.2 Hz, 1H), 4.75 - 4.64 (m, 1H), 4.49 - 4.41 (m, 1H), 4.16 - 3.92 (m, 2H), 3.90 - 3.81 (m, 4H), 3.70 (d, J = 9.3 Hz, 2H), 3.63 - 3.55 (m, 2H), 2.82 (s, 6H), 2.77 - 2.69 (m, 1H), 2.54 (t, J = 11.2 Hz, 1H), 1.98 - 1.83 (m, 5H), 1.71 (d, J= 13.3 Hz, 1H), 1.65 - 1.49 (m, 4H), 1.47 - 1.34 (m, 2H).MS (ESI) [M+H]+=403.3. Example 182 ( / f)-V-(2-(4-(dimethylamino)biit-2-eiioyl)isoindolin-5-yl)acetamide (XS209-55).

[0649] Example 182 was synthesized following similar procedure for preparing example 79. White solid, 39% yield 'H NMR (400 MHz, Methanol-d4) 8 7.55 (d, J = 21.6 Hz, 1H), 7.33 - 7.21 (m, 1H), 7.15 (t, J= 8.1 Hz, 1H), 6.81 - 6.61 (m, 2H), 4.85 (s, 2H), 4.66 (d, J= 7.2 Hz, 2H), 3.89 (d, J = 6.5 Hz, 2H), 2.82 (s, 6H), 2.01 (s, 3H).MS (ESI) [M+H]+=288.2.

[0650] Example 183 (£')-4-(dimethylamino)-l-(5-(methylamino)isoindolin-2-yl)but-2-en-l-one (XS209-57). Example 183 was synthesized following similar procedure for preparing example 79. White solid, 55% yieldJH NMR (400 MHz, Methanol-d)48 7.49 (t, J = 9.0 Hz, 1H), 7.42 (d, J = 7.1 Hz, 1H), 7.36 (d, J = 8.1 Hz, 1H), 6.84 (d, J= 15.2 Hz, 1H), 6.80 - 6.66 (m, 1H), 5.04 (d, J = 10.6 Hz, 2H), 4.83 (d, J = 8.7 Hz, 2H), 3.95 (d, J= 6.8 Hz, 2H), 3.03 (s, 3H), 2.87 (s, 6H).MS (ESI) [M+H]+=260.2.

[0651] Example 184 (E)-4-(dimethylamino)-l-(5-(methylamino)isoindolin-2-yl)but-2-en-l-one (XS209-60). Example 184 was synthesized following similar procedure for preparing example 79. White solid, 36% yield 'H NMR (400 MHz, Methanol-d4) 8 6.75 - 6.62 (m, 1H), 6.60 - 6.47 (m, 1H), 6.34 (s, 1H), 4.74 (d, J= 35.5 Hz, 1H), 4.50 (d, J= 33.5 Hz, 1H), 4.29 - 3.94 (m, 3H), 3.91 - 3.82 (m, 3H), 2.87 - 2.78 (m, 7H), 2.42 (d, J= 3.3 Hz, 3H), 1.68 (d, J= 7.1 Hz, 1H).MS (ESI) Example 185 (E)-l-(3-(6-acetyl-6-azaspiro[3.4]octane-2-carbonyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl)-4-(dimethylamino)but-2-en-l-one (XS209-62). Example 185 was synthesized following similar procedure for preparing example 79. White solid, 19% yield1H NMR (400 MHz, Methanol-d4) 8 6.72 - 6.58 (m, 1H), 6.47 (d, J= 15.2 Hz, 1H), 4.77 - 4.60 (m, 1H), 4.58 - 4.38 (m, 1H), 4.03 - 3.67 (m, 5H), 3.53 - 3.24 (m, 5H), 2.80 (s, 6H), 2.75 - 2.65 (m, 1H), 2.27 - 2.05 (m, 4H), 2.00 - 1.90 (m, 3H), 1.89 - 1.82 (m, 1H), 1.80 - 1.74 (m, 1H), 1.55 (d, J= 8.8 Hz, 1H), 1.46 - 1.40 (m, 1H).MS (ESI) [M+H]+=389.2.

[0652] Example 186 (E)-4-(dimethylamino)-l-(3-(5-methylthiazole-2-carbonyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl)but-2-en-l-one (XS209-65). Example 186 was synthesized following similar procedure for preparing example 79. White solid, 44% yield1H NMR (400 MHz, Methanol-^) 8 7.46 (d, J = 9.5 Hz, 1H), 6.65 - 6.50 (m, 1H), 6.49 - 6.34 (m, 1H), 4.65 (d, J = 14.6 Hz, 1H), 4.50 - 4.24 (m, 3H), 3.99 - 3.71 (m, 4H), 2.75 - 2.60 (m, 7H), 2.35 (s, 3H), 1.57 (d, J= 9.2 Hz, IH).MS (ESI) [M+H]1=335.1.

[0653] Example 187 8-(4-(dimethylamino)-4-methylpent-2-ynoyl)-3-(5-methylthiophen-2-yl)-3,8- diazabicyclo[3.2.1]octan-2-one (XS209-74). Example 187 was synthesized following similar procedure for preparing example 28. White solid, 29% yield 'H NMR (400 MHz, Methanol-d4) 8 6.49 (s, 2H), 4.99 (t, J= 5.5 Hz, 1H), 4.78 (d, J = 6.5 Hz, 1H), 4.04 - 3.90 (m, 1H), 3.73 - 3.54 (m, 1H), 2.97 - 2.88 (m, 6H), 2.40 - 2.20 (m, 5H), 2.18 - 2.08 (m, 1H), 2.04 - 1.89 (m, 1H), 1.77 - 1.64 (m, 6H).MS (ESI) [M+Hf =360.2.

[0654] Example 188 4-(dimethylamino)-4-methyl-l-(3-(5-methylthiophene-2-carbonyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl)pent-2-yn-l-one (XS209-75). Example 188 was synthesized following similar procedure for preparing example 79. White solid, 34% yield!H NMR (400 MHz, Methanol-^) 8 7.43 - 7.28 (m, 1H), 6.75 (t, J= 4.5 Hz, 1H), 4.71 - 4.47 (m, 2H), 4.40 - 4.20 (m, 1H), 4.20 - 4.06 (m, 1H), 4.06 - 3.78 (m, 2H), 3.13 - 3.00 (m, 1H), 2.92 - 2.85 (m, 8H), 2.74 (q, J= 7.1 Hz, 1H), 2.41 (s, 3H), 1.73 - 1.55 (m, 4H).MS (ESI) [M+H]+=360.1.

[0655] Example 189 (£)-4-(dimethylamino)-l-(3-(5-methyloxazole-2-carbonyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl)but-2-en-l-one (XS209-92). Example 189 was synthesized following similar procedure for preparing example 79. White solid, 29% yield 'H NMR (400 MHz, Methanol-d4) 8 6.89 (s, 1H), 6.66 - 6.54 (m, 1H), 6.52 - 6.38 (m, 1H), 4.73 - 4.63 (m, 1H), 4.48 - 4.22 (m, 3H), 3.94 - 3.72 (m, 4H), 2.77 - 2.66 (m, 7H), 2.26 (s, 3H), 1.59 (d, J= 9.2 Hz,

[0656] Example 190 l-(4-(dimethylamino)but-2-ynoyl)-3-hydroxy- / V-(5-methylthiophen-2- yl)azetidine-3-carboxamide (XS209-99). Example 190 was synthesized following similar procedure for preparing example 53. White solid, 29% yield1H NMR (400 MHz, Methanol-d4) 8 6.55 (d, J= 3.4 Hz, 1H), 6.41 (d, J= 4.2 Hz, 1H), 4.55 (d, J= 9.7 Hz, 1H), 4.31 (d, J= 11.0 Hz, 1H), 4.23 (s, 2H), 4.13 (d, J= 9.7 Hz, 1H), 3.90 (d, J= 10.9 Hz, 1H), 2.88 (s, 6H), 2.28 (s, 3H).

[0657] MS (ESI) [M+H]+=322.1.

[0658] Example 193 (E)-8-(4-(dimethylamino)but-2-enoyl)-N-(5-methylthiophen-2-yl)-8- azabicyclo[3.2.1]octane-3-carboxamide (XS209-122). Example 193 was synthesized following similar procedure for preparing example 53. White solid1H NMR (400 MHz, Methanol-d4) 8 6.79 (d, J= 15.2 Hz, 1H), 6.72 - 6.60 (m, 1H), 6.38 (s, 2H), 4.62 (d, J= 6.6 Hz, 1H), 4.52 - 4.40 (m, 1H), 3.86 (d, J= 7.0 Hz, 2H), 3.03 - 2.90 (m, 1H), 2.82 (s, 6H), 2.26 (s, 3H), 2.13 - 1.99 (m, 1H), 1.98 - 1.59 (m, 7H). MS (ESI) [M+H]+=362.2.

[0659] Example 194 (£)-JV-(8-(4-(dimethylamino)but-2-enoyl)-8-azabicyclo[3.2.1]octan-3-yl)-5- methylthiophene-2-carboxamide (XS209-139). Example 194 was synthesized following similar procedure for preparing example 79. White solid, 45% yield 'H NMR (400 MHz, Methanol-6 / 4) 6 7.34 (d, J= 3.2 Hz, 1H), 6.80 - 6.51 (m, 3H), 4.51 (s, 1H), 4.37 (s, 1H), 3.89 (s, 1H), 3.81 (d, J = 6.8 Hz, 2H), 2.76 (s, 6H), 2.35 (s, 3H), 2.15 - 2.05 (m, 1H), 2.05 - 1.91 (m, 5H), 1.90 - 1.76 (m,

[0660] Example 196 (E)-W-(4-(4-(dimethylamino)but-2-enamido)phenyl)-5-methylthiophene-2- carboxamide (XS209-165). Example 196 was synthesized following similar procedure for preparing example 79. White solid, 33% yield1H NMR (400 MHz, Methanol-d4) 8 7.71 (d, J = 3.5 Hz, 1H), 7.65 (s, 4H), 6.93 - 6.76 (m, 2H), 6.56 (d, J= 15.2 Hz, 1H), 4.00 (d, J = 7.2 Hz, 2H), 2.94 (s, 6H), 2.55 (s, 3H). MS (ESI) [M+H]+=344.1.

[0661] Example 197 l-(3-butyryl-3,6-diazabicyclo[3.1.1]heptan-6-yl)-4-(dimethylamino)but-2-yn- 1-one (XS209-166). Example 197 was synthesized following similar procedure for preparing example 79. White solid, 18% yield 'H NMR (400 MHz, Methanol-d)48 4.62 (s, 1H), 4.59 - 4.47 (m, 1H), 4.27 (s, 2H), 4.04 - 3.93 (m, 1H), 3.87 - 3.77 (m, 1H), 3.77 - 3.62 (m, 2H), 2.94 - 2.90 (m, 6H), 2.79 - 2.64 (m, 1H), 2.40 - 2.20 (m, 2H), 1.64 - 1.52 (m, 3H), 0.97 - 0.88 (m, 3H).MS (ESI) [M+H]+=278.2.

[0662] Example 198 l-(3-acetyl-3,6-diazabicyclo[3.1.1]heptan-6-yl)-4-(dimethylamino)-4- methylpent-2-yn-l-one (XS209-167). Example 198 was synthesized following similar procedure for preparing example 79. White solid, 19% yield 'H NMR (400 MHz, Methanol-d)48 4.66 (d, J = 6.2 Hz, 2H), 4.10 (d, J= 12.0 Hz, 1H), 3.94 (d, J= 13.6 Hz, 1H), 3.86 (d, J= 12. 1 Hz, 1H), 3.75 (d, J= 13.6 Hz, 1H), 3.15 - 3.04 (m, 1H), 2.93 (s, 6H), 2.92 (s, 6H), 2.12 (s, 3H), 1.81 (d, J= 9.0 Hz, 1H). MS (ESI) [M+H]+=278.2.

[0663] Example 199 1 -(3-butyryl-3,6-diazabicyclo[3.1.1 ]heptan-6-yl)-4-(dimethylamino)-4- methylpent-2-yn-l-one (XS209-168). Example 199 was synthesized following similar procedure for preparing example 79. White solid, 20% yield 'H NMR (400 MHz, Methanol-d)48 4.64 (d, J = 5.8 Hz, 2H), 4.06 (d, J= 11.9 Hz, 1H), 3.91 (d, J= 13.5 Hz, 1H), 3.84 (d, J= 12. 1 Hz, 1H), 3.72 (d, J= 13.4 Hz, 1H), 3.07 (q, .7= 7.4 Hz, 1H), 2.92 - 2.86 (m, 12H), 2.40 - 2.29 (m, 2H), 1.76 (d, J= 7.9 Hz, 1H), 1.63 - 1.56 (m, 2H), 0.92 (t, J= 7.3 Hz, 3H). MS (ESI) [M+H]+=306.2.

[0664]

[0665] Example 200: l-(3-butyryl-3,6-diazabicyclo[3.1.1]heptan-6-yl)-4-(dimethylamino)-4- methylpent-2-yn-l-one (XS209-174). Example 200 was synthesized following similar procedure for preparing example 53. White solid, 45% yield 'H NMR (400 MHz, Methanol-d)45 7.66 (d, J

[0666] = 7.8 Hz, 1H), 7.55 (d, J= 7.8 Hz, 1H), 7.21 (t, J= 7.5 Hz, 1H), 7.14 (t, J = 7.4 Hz, 1H), 7.06 (s,

[0667] 1H), 6.74 - 6.61 (m, 1H), 6.47 (d, J= 15.2 Hz, 1H), 4.66 (d, J= 9.5 Hz, 1H), 4.40 (d, J= 11.0 Hz, 1H), 4.23 (d, J= 9.5 Hz, 1H), 3.98 (d, J= 11.0 Hz, 1H), 3.87 (d, J = 7.1 Hz, 2H), 2.82 (s, 6H). MS

[0668] Example 201: N-(benzo[b]thiophen-2-yl)-l-(4-(dimethylamino)but-2-ynoyl)-3- methylazetidine-3-carboxamide (XS209-175). Example 201 was synthesized following similar procedure for preparing example 53. White solid, 44% yield *HNMR (400 MHz, Methanol-d4) δ 7.67 (d, J= 7.9 Hz, 1H), 7.55 (d, J= 7.8 Hz, 1H), 7.22 (t, J= 7.4 Hz, 1H), 7.15 (t, J = 7.4 Hz, 1H), 6.94 (s, 1H), 4.61 (d, J= 9.2 Hz, 1H), 4.32 (d, J= 10.6 Hz, 1H), 4.27 (s, 2H), 4.05 (d, J= 9.2 Hz, 1H), 3.87 (d, J= 10.6 Hz, 1H), 2.93 (s, 6H), 1.63 (s, 3H).MS (ESI) [M+Hf =345.2. Example 202: (E)-l-(4-(dimethylamino)but-2-enoyl)-3-hydroxy-iV-(4, 5,6,7- tetrahydrobenzo[Z>]thiophen-2-yl)azetidine-3-carboxamide (XS209-176). Example 202 was synthesized following similar procedure for preparing example 53. White solid, 39% yield 'H NMR (400 MHz, Methanol-d4) 8 6.70 - 6.55 (m, 1H), 6.49 - 6.36 (m, 2H), 4.58 (d, J= 9.4 Hz, 1H), 4.32 (d, J= 10.9 Hz, 1H), 4.16 (d, J = 9.4 Hz, 1H), 3.91 (d, J= 10.9 Hz, 1H), 3.84 (d, J= 1A Hz, 2H), 2.79 (s, 6H), 2.54 (t, J= 62 Hz, 2H), 2.40 (t, J= 5.9 Hz, 2H), 1.79 - 1.59 (m, 4H). MS (ESI) [M+H]+=364.2.

[0669] Example 203: (E)-l-(3-(5-(4-acetylpiperazin-l-yl)thiophene-2-carbonyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl)-4-(dimethylamino)but-2-en-l-one (XS209-184). Example 203 was synthesized following similar procedure for preparing example 79. White solid, 28% yield

[0670] NMR (400 MHz, Methanol-d4) δ 7.46 (s, 1H), 6.84 - 6.69 (m, 1H), 6.60 (d, J= 15.3 Hz, 1H), 6.25 (d, J = 4.1 Hz, 1H), 4.82 (s, 1H), 4.60 (s, 1H), 4.02 - 3.87 (m, 3H), 3.79 - 3.63 (m, 4H), 3.38 - 3.30 (m, 7H), 2.91 (s, 6H), 2.87 - 2.79 (m, 1H), 2.17 (s, 3H), 1.74 (d, J= 9.1 Hz, 1H). MS (ESI) [M+H]1=446.2

[0671] Example 204: (E)-4-(dimethylamino)-l-(3-(5-(4-methylpiperazin-l-yl)thiophene-2- carbonyl)-3,6-diazabicyclo[3.1.1]heptan-6-yl)but-2-en-l-one (XS209-185). Example 204 was synthesized following similar procedure for preparing example 79. White solid, 23% yield 'H NMR (400 MHz, Methanol-d4) 8 7.32 (s, 1H), 6.70 - 6.58 (m, 1H), 6.47 (d, J= 15.3 Hz, 1H), 6.20 (d, J= 4.1 Hz, 1H), 4.69 (s, 1H), 4.46 (s, 1H), 4.37 - 3.89 (m, 3H), 3.83 (d, J= 7.0 Hz, 3H), 3.73 (d, J = 9.8 Hz, 2H), 3.57 - 3.39 (m, 2H), 3.19 - 3.07 (m, 4H), 2.85 (s, 3H), 2.77 (s, 6H), 2.74 - 2.65 (m, 1H), 1.60 (d, J= 9.1 Hz, 1H).MS (ESI) [M+H]+=418.2

[0672] Example 205: (E)-JV-(4-(4-(dimethylamino)but-2-enamido)cyclohexyl)-5-methylthiophene- 2-carboxamide (XS224-6). Example 205 was synthesized following similar procedure for preparing example 79. White solid, 25% yield1H NMR (400 MHz, Methano)l 8-d 74.53 - 7.31 (m, 1H), 6.73 (s, 1H), 6.70 - 6.53 (m, 1H), 6.29 (d, J= 15.3 Hz, 1H), 3.87 (d, J= 6.7 Hz, 2H), 3.84 - 3.62 (m, 2H), 2.84 (s, 6H), 2.44 (s, 3H), 1.94 (d, J= 9.3 Hz, 4H), 1.57 - 1.22 (m, 4H).MS (ESI)

[0673] Example 206: 2V-(benzo[ / >]thiophen-2-yl)-l-(4-(dimethylamino)but-2-ynoyl)azetidine-3- carboxamide (XS224-106). Example 206 was synthesized following similar procedure for preparing example 53. White solid, 44% yield *H NMR (400 MHz, Methanol-d4) 87.72 (d, J= 7.8 Hz, 1H), 7.61 (d, J= 7.8 Hz, 1H), 7.28 (t, J= 7.5 Hz, 1H), 7.21 (t, J= 7.5 Hz, 1H), 6.95 (s, 1H), 4.49 (p, J= 9.3 Hz, 2H), 4.31 (d, J= 13.3 Hz, 3H), 4.25 - 4.17 (m, 1H), 3.67 (p, J= 7.1, 6.0 Hz, 1H), 2.98 (s, 6H). MS (ESI) [M+H]+=342.1.

[0674] Example 207: / V-(benzo|6]thiophen-2-yl)-l-(4-(dimethylamino)-4-methylpent-2- ynoyl)azetidine-3-carboxamide (XS224-107). Example 207 was synthesized following similar procedure for preparing example 53. White solid, 39% yield1H NMR (400 MHz, Methanol)-d 847.72 (d, J= 7.8 Hz, 1H), 7.60 (d, J= 7.8 Hz, 1H), 7.28 (t, J= 7.5 Hz, 1H), 7.20 (t, J = 7.4 Hz, 1H), ΰ.95 (s, 1H), 4.48 (p, J= 8.7 Hz, 2H), 4.29 (t, J= 9.6 Hz, 1H), 4.24 - 4.16 (m, 1H), 3.67 (p, J = 7.2, 6.0 Hz, 1H), 2.98 (s, 6H), 1.75 (s, 6H). MS (ESI) [M+H]+=370.2.

[0675] Example 208: l-(4-(dimethylamino)but-2-ynoyl)- / V-(4,5,6,7-tetrahydrobenzo[Z>]thiophen-2- yl)azetidine-3-carboxamide (XS224-108). Example 208 was synthesized following similar procedure for preparing example 53. White solid, 38% yield 'H NMR (400 MHz, Methanol-d4) δ 6.33 (s, 1H), 4.48 - 4.35 (m, 2H), 4.29 (s, 2H), 4.25 - 4.17 (m, 1H), 4.14 - 4.08 (m, 1H), 3.62 - 3.51 (m, 1H), 2.95 (s, 6H), 2.60 (t, J= 6.3 Hz, 2H), 2.45 (t, J= 5.7 Hz, 2H), 1.85 - 1.67 (m, 4H). MS (ESI) [M+H]+=346.2.

[0676] Example 210: l-(4-(dimethylamino)but-2-ynoyl)-Az-(5-methylthiophen-2-yl)azetidine-3- carboxamide (XS224-110). Example 210 was synthesized following similar procedure for preparing example 53. White solid, 29% yield 'H NMR (400 MHz, Methanol-d4) 8 6.47 (s, 2H), 4.42 (p, J = 9.1 Hz, 2H), 4.31 (s, 2H), 4.23 (t, J = 9.7 Hz, 1H), 4.17 - 4.09 (m, 1H), 3.64 - 3.50 (m, 1H), 2.96 (s, 6H), 2.35 (s, 3H).MS (ESI) [M+H]+=306.1.

[0677] Example 212: l-(4-(dimethylamino)biit-2-ynoyl)- V-(5-isopropylthiophen-2-yl)azetidine-3- carboxamide (XS224-116). Example 212 was synthesized following similar procedure for preparing example 53. White solid, 35% yield1H NMR (400 MHz, Methano)l 8-d 64.51 (d, J= 5.6 Hz, 2H), 4.43 (p, J= 8.9 Hz, 2H), 4.31 (s, 2H), 4.24 (t, J= 9.6 Hz, 1H), 4.14 (d, J= 15.8 Hz, 1H), 3.58 (p, J= 7.0, 6.2 Hz, 1H), 3.09 - 3.00 (m, 1H), 2.96 (s, 6H), 1.27 (d, J= 6.8 Hz, 6H). MS (ESI) [M+H]+=334.2.

[0678] Example 213: l-(4-(dimethylamino)-4-methylpent-2-ynoyl)- / V-(5-isopropylthiophen-2- yl)azetidine-3-carboxamide (XS224-117). Example 213 was synthesized following similar procedure for preparing example 53. White solid, 25% yield 'H NMR (400 MHz, Methanol-d)48 6.51 (d, J= 5.9 Hz, 2H), 4.50 - 4.34 (m, 2H), 4.24 (t, J = 9.6 Hz, 1H), 4.18 - 4.11 (m, 1H), 3.58 (p, J = 7.2, 6.0 Hz, 1H), 3.11 - 3.00 (m, 1H), 2.97 (s, 6H), 1.74 (s, 6H), 1.27 (d, J= 6.8 Hz, 6H). MS (ESI) [M+H]+=362.1.

[0679] Example 216:(E')-4-(dimethylamino)-N -(l-(5-methylthiophene-2-carbonyl)azetidin-3- yl)but-2-enamide (XS224-143). Example 216 was synthesized following similar procedure for preparing example 79. White solid, 33% yield ’H NMR (400 MHz, Methanol-d4) 87.29 (d, J= 3.1 Hz, 1H), 6.78 (s, 1H), 6.74 - 6.59 (m, 1H), 6.31 (d, J= 15.3 Hz, 1H), 4.82 - 4.61 (m, 2H), 4.35 (d, J = 38.0 Hz, 2H), 3.98 (s, 1H), 3.88 (d, J = 7.2 Hz, 2H), 2.83 (s, 6H), 2.45 (s, 3H). MS (ESI) [M+H]+=308.2.

[0680] Example 217: 4-(dimethylamino)-2V-(l-(5-methylthiophene-2-carbonyl)azetidin-3-yl)but-2- ynamide (XS224-144). Example 217 was synthesized following similar procedure for preparing example 79. White solid, 29% yield 'HNMR (400 MHz, Methanol-d)45 7.29 (d, J= 3.2 Hz, 1H), 6.79 (s, 1H), 4.81 - 4.60 (m, 2H), 4.47 - 4.20 (m, 4H), 3.99 (s, 1H), 2.94 (s, 6H), 2.45 (s, 3H). MS (ESI) [M+H]+=306.2.

[0681] Example 219: (E)-W-(3-(4-(dimethylamino)but-2-enamido)cyclobutyl)-5-methylthiophene-2- carboxamide (XS224-147). Example 219 was synthesized following similar procedure for preparing example 79. White solid, 45% yield1H NMR (400 MHz, Methanol-d4) 8 7.48 - 7.35 (m, 1H), 6.71 (s, 1H), 6.69 - 6.54 (m, 1H), 6.34 - 6.20 (m, 1H), 4.46 (p, J= 6.9, 6.4 Hz, 1H), 4.38 - 4.26 (m, 1H), 3.89 - 3.77 (m, 2H), 2.80 (s, 6H), 2.73 - 2.61 (m, 1H), 2.47 - 2.35 (m, 4H), 2.35 - 2.25 (m, 1H), 2.08 - 1.91 (m, 1H). MS (ESI) [M+H]+=322.1. Example 223: (£)-N-(l-(4-(dimethylamino)but-2-enoyl)azetidin-3-yl)benzo[6]thiophene-2- carboxamide (XS224-154). Example 223 was synthesized following similar procedure for preparing example 79. White solid, 36% yield 'H NMR (400 MHz, Methanol-d)45 7.98 (s, 1H), 7.88 (t, J= 6.9 Hz, 2H), 7.42 (p, J= 6.9 Hz, 2H), 6.77 - 6.62 (m, 1H), 6.50 (d, J= 15.3 Hz, 1H), 4.83 - 4.76 (m, 1H), 4.67 (t, J= 8.5 Hz, 1H), 4.42 (t, J= 9.5 Hz, 1H), 4.37 - 4.29 (m, 1H), 4.19 - 4.11 (m, 1H), 3.93 (d, J= 7.0 Hz, 2H), 2.88 (s, 6H). MS (ESI) [M+H]+=344.2.

[0682] Example 224: JV-(l-(4-(dimethylamino)but-2-ynoyl)azetidin-3-yl)benzo[ / >]thiophene-2- carboxamide (XS224-155). Example 224 was synthesized following similar procedure for preparing example 79. White solid, 29% yield 'H NMR (400 MHz, Methanol-d)48 7.99 (s, 1H), 7.90 (t, J = 6.4 Hz, 2H), 7.43 (p, J = 7.0 Hz, 2H), 4.84 - 4.78 (m, 1H), 4.66 (t, J= 8.7 Hz, 1H), 4.42 (t, , / = 9.5 Hz, 1H), 4.37 - 4.29 (m, 3H), 4.20 - 4.08 (m, 1H), 2.99 (s, 6H). MS (ESI) [M+H]+=342.2.

[0683] Example 225: 7V-(l-(4-(dimethylamino)-4-methylpent-2-ynoyl)azetidin-3- yl)benzo[ / >]thiophene-2-carboxamide (XS224-156). Example 225 was synthesized following similar procedure for preparing example 79. White solid, 26% yield 'H NMR (400 MHz, Methanol-^) 87.99 (s, 1H), 7.90 (t, J= 6.6 Hz, 2H), 7.43 (p, J= 6.7 Hz, 2H), 4.83 - 4.76 (m, 1H), 4.65 (t, J= 8.7 Hz, 1H), 4.42 (t, J= 9.5 Hz, 1H), 4.36 - 4.30 (m, 1H), 4.20 - 4.09 (m, 1H), 2.99 (s, 6H), 1.76 (s, 6H). MS (ESI) [M+H]+=370.2. Example 226: (E)-2V-(1-(4-(dimethylamino)but-2-enoyl)azetidin-3-yl)-4, 5,6,7- tetrahydrobenzo[Z>]thiophene-2-carboxamide (XS224-157). Example 226 was synthesized following similar procedure for preparing example 79. White solid, 25% yieldXH NMR (400 MHz, Methanol-d4) 8 7.31 (s, 1H), 6.71 - 6.57 (m, 1H), 6.43 (d, J= 15.3 Hz, 1H), 4.73 - 4.62 (m, 1H), 4.58 (t, J= 8.5 Hz, 1H), 4.32 (t, J = 9.4 Hz, 1H), 4.26 - 4.16 (m, 1H), 4.09 - 3.97 (m, 1H), 3.87 (d, J= 7.0 Hz, 2H), 2.82 (s, 6H), 2.69 (t, J= 6.3 Hz, 2H), 2.53 (t, J= 5.8 Hz, 2H), 1.81 - 1.64 (m, 4H). MS (ESI) [M+H]+=348.3.

[0684] Example 227: 2V-(l-(4-(dimethylamino)but-2-ynoyl)azetidin-3-yl)-4, 5,6,7- tetrahydrobenzo[Z>]thiophene-2-carboxamide (XS224-158). Example 227 was synthesized following similar procedure for preparing example 79. White solid, 23% yield1H NMR (400 MHz, Methanol-d4) 8 7.37 (s, 1H), 4.79 - 4.68 (m, 1H), 4.60 (t, ,7 = 8.7 Hz, 1H), 4.40 - 4.30 (m, 3H), 4.30 - 4.21 (m, 1H), 4.14 - 4.02 (m, 1H), 2.98 (s, 6H), 2.77 (t, J= 6.0 Hz, 2H), 2.61 (t, .7= 6.0 Hz, 2H), 1.93 - 1.72 (m, 4H). MS (ESI) [M+H]+=346.2.

[0685] Example 228: / V-(l-(4-(dimethylamino)-4-methylpent-2-ynoyl)azetidin-3-yl)-4, 5,6,7- tetrahydrobenzo[b]thiophene-2-carboxamide (XS224-159). Example 228 was synthesized following similar procedure for preparing example 79. White solid, 29% yield1H NMR (400 MHz, Methanol-d4) 8 7.36 (s, 1H), 4.74 (p, J= 6.5, 5.9 Hz, 1H), 4.59 (t, J= 8.6 Hz, 1H), 4.36 (t, J = 9.5 Hz, 1H), 4.29 - 4.20 (m, 1H), 4.12 - 4.03 (m, 1H), 2.98 (s, 6H), 2.75 (t, J= 6.4 Hz, 2H), 2.60 (t, J = 6.0 Hz, 2H), 1.89 - 1.77 (m, 4H), 1.75 (s, 6H). MS (ESI) [M+H]+=374.2.

[0686] Procedures for the synthesis of AMPK based bivalent compounds

[0687]

[0688] Linker 10-14: n = 1-5

[0689] The linkers 1-14 were synthesized following the reported procedures (Liu et al., 2022)

[0690] Scheme 12. Syntheses of example 229 example 229

[0691] Example 229: A-(2-(3-(5-(4-acryloyl-2-oxopiperazin-l-yl)furan-2-yl)propanamido)ethyl)-5- ((6-chloro-5-(l-methyl-l / 7-indol-5-yl)-l / 7-benzo[c / ]imidazol-2-yl)oxy)-2- methylbenzamide(XF137-81). To a solution of activator 991 (prepared according to previous published paper) (Ngoei et al., 2018) (21 mg, 0.048 mmol) in DMSO (1 mL) were added linker 1 (16.3 mg, 0.048 mmol, 1.0 equiv), EDCI (14 mg, 0.072 mmol, 1.5 equiv), HOAt (10 mg, 0.072 mmol, 1.5 equiv), and NMM (15 mg, 0.14 mmol, 3.0 equiv). After being stirred for 3 h at rt, the resulting mixture was purified by preparative HPLC to afford titled compound as brown solid. (6.7 mg, yield 19%).1H NMR (400 MHz, CD3OD) δ 7.62 - 7.48 (m, 3H), 7.48 - 7.35 (m, 5H), 7.27 - 7.18 (m, 2H), 6.75 - 6.59 (m, 1H), 6.30 - 6.25 (m, 1H), 6.16 (d, J= 3.1 Hz, 1H), 6.00 (d, J= 3.2 Hz, 1H), 5.80 (d, J= 10.5 Hz, 1H), 4.42 - 4.21 (m, 2H), 3.91 - 3.82 (m, 5H), 3.81 - 3.69 (m, 2H), 3.53 - 3.38 (m, 4H), 2.80 (t, J= 7.5 Hz, 2H), 2.47 (s, 5H). MS (ESI) [M+H]+= 748.3.

[0692] Example 230: JV-(3-(3-(5-(4-acryloyl-2-oxopiperazin-l-yl)furan-2-yl)propanamido)propyl)- 5-((6-chloro-5-( 1-methyl- l / / -indol-5-yl)- 1H-benzo[d / ]imidazol-2-yl)oxy)-2-methylbenzamide (XF137-82). Example 230 was synthesized following the standard procedure for preparing example 229 from activator 991 (28 mg, 0.06 mmol) and linker 2 (23 mg, 0.06 mmol, 1.0 equiv). Brown solid (15.4 mg, yield 34%). ’H NMR (400 MHz, CD3OD) δ 7.61 - 7.47 (m, 3H), 7.45 - 7.34 (m, 5H), 7.28 - 7.14 (m, 2H), 6.83 - 6.61 (m, 1H), 6.28 (dd, J= 16.8, 1.9 Hz, 1H), 6.21 (d, J = 3.2 Hz, 1H), 6.09 (d, J= 3.2 Hz, 1H), 5.80 (dd, J= 10.5, 1.9 Hz, 1H), 4.45 - 4.27 (m, 2H), 3.93 (t, J= 5.3 Hz, 2H), 3.84 (d, J= 9.3 Hz, 5H), 3.38 - 3.34 (m, 2H), 3.27 (d, J= 6.8 Hz, 2H), 2.88 (t, J= 7.3 Hz, 2H), 2.48 (d, J= 11.3 Hz, 5H), 1.76 (t, J= 6.8 Hz, 2H). MS (ESI) [M+H]+= 762.3.

[0693] Example 231: 7V-(4-(3-(5-(4-acryloyl-2-oxopiperazin-l-yl)furan-2-yl)propanamido)butyl)-5- ((6-chloro-5-(l-methyl-l / / -indol-5-yl)-lH -benzo[<7|imidazol-2-yl)oxy)-2-methylbenzamide

[0694] (XF137-83). Example 231 was synthesized following the standard procedure for preparing example 229 from activator 991 (21 mg, 0.048 mmol) and liner 3 (17 mg, 0.048 mmol, 1.0 equiv). Brown solid (7.6 mg, yield 20%). 'HNMR (400 MHz, CD3OD) 5 7.57 (d, J= 2.5 Hz, 3H), 7.51 - 7.32 (m, 5H), 7.23 (d, J= 8.3 Hz, 2H), 6.84 - 6.57 (m, 1H), 6.29 (dd, J = 16.8, 1.9 Hz, 1H), 6.20 (d, J= 3.2 Hz, 1H), 6.08 (d, J= 3.2 Hz, 1H), 5.81 (dd, J= 10.6, 1.9 Hz, 1H), 4.44 - 4.24 (m, 2H), 3.95 - 3.89 (m, 2H), 3.87 - 3.75 (m, 5H), 3.47 - 3.35 (m, 2H), 3.21 (t, J= 6.4 Hz, 2H), 2.87 (t, J = 7.3 Hz, 2H), 2.47 (s, 5H), 1.73 - 1.47 (m, 4H). MS (ESI) [M+H]+= 776.3.

[0695]

[0696] Example 233: JV-(6-(3-(5-(4-acryloyl-2-oxopiperazin-l-yl)furan-2-yl)propanamido)hexyl)-5- ((6-chloro-5-(l-methyl-lH -indol-5-yl)-lH-benzo[d]imidazol-2-yl)oxy)-2-methylbenzamide (XF137-85). Example 233 was synthesized following the standard procedure for preparing example 229 from activator 991 (21 mg, 0.048 mmol) and linker 5 (18.7 mg, 0.048 mmol, 1.0 equiv). Compound XF137-85 was obtained as brown solid (9.1 mg, yield 24%). 'H NMR (400 MHz, CD3OD) δ 7.58 (d, J= 3.0 Hz, 2H), 7.52 - 7.32 (m, 6H), 7.23 (d, J= 8.3 Hz, 2H), 6.85 - 6.61 (m, 1H), 6.29 (dd, J= 16.7, 2.0 Hz, 1H), 6.22 (d, J= 3.2 Hz, 1H), 6.08 (d, J= 3.2 Hz, 1H), 5.82 (dd, J = 10.5, 1.9 Hz, 1H), 4.47 - 4.26 (m, 2H), 4.03 - 3.89 (m, 2H), 3.89 - 3.74 (m, 5H), 3.43 - 3.34 (m, 2H), 3.15 (t, J= 6.8 Hz, 2H), 2.88 (t, J= 7.3 Hz, 2H), 2.47 (s, 5H), 1.69 - 1.55 (m, 2H), 1.54 - 1.27 (m, 6H). MS (ESI) [M+H]+= 804.3.

[0697] Example 234: 2V-(7-(3-(5-(4-acryloyl-2-oxopiperazin-l-yl)furan-2-yl)propanamido)heptyl)- 5-((6-chloro-5-(l-methyl-l / / -indol-5-yl)-l / / -benzo[t / |imidazol-2-yl)oxy)-2-methylbenzamide (XF137-86). Example 234 was synthesized following the standard procedure for preparing example 229 from activator 991 (21 mg, 0.048 mmol) and linker 6 (19 mg, 0.048 mmol, 1.0 equiv). Brown solid (12.6 mg, yield 32%). ’H NMR (400 MHz, CD3OD) 8 7.63 - 7.52 (m, 2H), 7.49 - 7.31 (m, 6H), 7.28 - 7.16 (m, 2H), 6.88 - 6.64 (m, 1H), 6.30 (dd, J= 16.8, 1.9 Hz, 1H), 6.22 (d, J = 3.3 Hz, 1H), 6.08 (d, 7= 3.2 Hz, 1H), 5.82 (dd, J= 10.6, 1.9 Hz, 1H), 4.47 - 4.31 (m, 2H), 4.06 - 3.94 (m, 2H), 3.86 (s, 5H), 3.41 - 3.36 (m, 2H), 3.14 (t, J= 6.9 Hz, 2H), 2.89 (t, J= 7.3 Hz, 2H), 2.52 - 2.40 (m, 5H), 1.62 (q, J= 7.1 Hz, 2H), 1.52 - 1.21 (m, 8H). MS (ESI) [M+H]+= 818.4.

[0698] Example 235: JV-(8-(3-(5-(4-acryloyl-2-oxopiperazin-l-yl)furan-2-yl)propanamido)octyl)-5- ((6-chloro-5-(l-methyl-17 / -indol-5-yl)-177-benzo[7]imidazol-2-yl)oxy)-2-methylbenzamide (XF137-87). Example 235 was synthesized following the standard procedure for preparing Example 229 from activator 991 (21 mg, 0.048 mmol) and linker 7 (20 mg, 0.048 mmol, 1.0 equiv). Brown solid (9.6 mg, yield 24%). 'H NMR (400 MHz, CD3OD) 8 'H NMR (400 MHz, MeOD) 8 7.62 - 7.52 (m, 2H), 7.49 - 7.36 (m, 6H), 7.28 - 7.18 (m, 2H), 6.82 - 6.64 (m, 1H), 6.35 - 6.26 (m, 1H), 6.25 - 6.21 (m, 1H), 6.08 (d, J= 3.2 Hz, 1H), 5.82 (dt, J= 10.6, 1.3 Hz, 1H), 4.40 (d, J = 26.3 Hz, 2H), 4.02 - 3.92 (m, 2H), 3.89 - 3.78 (m, 5H), 3.42 - 3.35 (m, 2H), 3.13 (t, J= 7.0 Hz, 2H), 2.89 (t, 7= 7.3 Hz, 2H), 2.55 - 2.39 (m, 5H), 1.62 (q, 7= 7.2 Hz, 2H), 1.52 - 1.24 (m, 10H). MS (ESI) [M+H]+= 832.2.

[0699] Example 236: JV-(9-(3-(5-(4-acryloyl-2-oxopiperazin-l-yl)furan-2-yl)propanamido)nonyl)-5- ((6-chloro-5-(l-methyl-177-indol-5-yl)-177-benzo[<7|imidazol-2-yl)oxy)-2-methylbenzamide (XF137-88). Example 236 was synthesized following the standard procedure for preparing example 229 from activator 991 (21 mg, 0.048 mmol) and linker 8 (21 mg, 0.048 mmol, 1.0 equiv). Brown solid (12.9 mg, yield 32%). 'H NMR (400 MHz, CD3OD) 6 7.54 (d, J= 17.1 Hz, 2H), 7.43 - 7.28 (m, 6H), 7.28 - 7.10 (m, 2H), 6.73 (d, J= 10.8 Hz, 1H), 6.30 - 6.28 (m, 1H), 6.22 (d, J = 3.2 Hz, 1H), 6.11 - 5.98 (m, 1H), 5.81 (dd, J= 10.5, 2.0 Hz, 1H), 4.46 - 4.26 (m, 2H), 3.93 (t, J = 5.4 Hz, 2H), 3.87 - 3.76 (m, 5H), 3.41 - 3.34 (m, 2H), 3.12 (t, ,7 = 7.0 Hz, 2H), 2.88 (t, ,7= 7.3 Hz, 2H), 2.51 - 2.38 (m, 5H), 1.61 (q, J = 7.2 Hz, 2H), 1.50 - 1.22 (m, 12H). MS (ESI) [M+H]+=

[0700] 846.4.

[0701] Example 237 : 2V-(10-(3-(5-(4-acryloyl-2-oxopiperazin-l-yl)furan-2-yI)propanamido)decyl)- 5-((6-chloro-5-(l-methyl-17 / -indol-5-yl)-177-benzo[t / ]imidazol-2-yl)oxy)-2-methylbenzamide (XF137-89). Example 237 was synthesized following the standard procedure for preparing example 229 from activator 991 (21 mg, 0.048 mmol) and linker 9 (21 mg, 0.048 mmol, 1.0 equiv). Brown solid (12.8 mg, yield 31%). ’H NMR (400 MHz, CD3OD) 8 7.62 - 7.47 (m, 2H), 7.47 - 7.29 (m, 6H), 7.27 - 7.19 (m, 2H), 6.82 - 6.65 (m, 1H), 6.35 - 6.25 (m, 1H), 6.24 - 6.17 (m, 1H), 6.08 (d, .7= 3.2 Hz, 1H), 5.88 - 5.67 (m, 1H), 4.48 - 4.28 (m, 2H), 3.94 (t, J= 5.1 Hz, 2H), 3.84 (d, J= 1.4 Hz, 5H), 3.40 - 3.34 (m, 2H), 3.12 (t, J= 7.0 Hz, 2H), 2.89 (t, J= 7.3 Hz, 2H), 2.46 (d, J= 1.8 Hz, 5H), 1.62 (q, J= 7.1 Hz, 2H), 1.50 - 1.17 (m, 14H). MS (ESI) [M+H]+= 860.3.

[0702] Example 238: 2V-(2-(2-(3-(5-(4-acryloyl-2-oxopiperazin-l-yl)furan-2- yl)propanamido)ethoxy)ethyl)-5-((6-chloro-5-(l-methyl-l / 7-indol-5-yl)-l / / - benzo [d / |imidazol-2-yl)oxy)-2-ni ethylbenzamide (XF137-90). Example 238 was synthesized following the standard procedure for preparing example 229 from activator 991 (21 mg, 0.048 mmol) and linker 10 (18.1 mg, 0.048 mmol, 1.0 equiv). Brown solid (16 mg, yield 42%). 'H NMR (400 MHz, CD3OD) δ 7.60 - 7.52 (m, 2H), 7.51 - 7.36 (m, 6H), 7.26 - 7.18 (m, 2H), 6.82 - 6.59 (m, 1H), 6.28 (dd, J= 16.8, 1.9 Hz, 1H), 6.20 (d, J= 3.2 Hz, 1H), 6.03 (d, J= 3.2 Hz, 1H), 5.80

[0703] (dd, J= 10.6, 1.9 Hz, 1H), 4.41 - 4.26 (m, 2H), 3.90 (t, J= 5.4 Hz, 2H), 3.85 (s, 3H), 3.78 (s, 2H),

[0704] 3.67 - 3.49 (m, 6H), 3.38 - 3.35 (m, 2H), 2.79 (t, J= 1A Hz, 2H), 2.47 (s, 3H), 2.42 (t, J= 7.4 Hz,

[0705] 2H). MS (ESI) [M+H]+= 792.2.

[0706] Example 239: JV-(2-(2-(2-(3-(5-(4-acryloyl-2-oxopiperazin-l-yl)furan-2- yl)propanamido)ethoxy)ethoxy)ethyl)-5-((6-chloro-5-(l-methyl-l / 7-indol-5-yl)-EH- benzo[d]imidazol-2-yl)oxy)-2-methylbenzamide (XF137-91). Example 239 was synthesized following the standard procedure for preparing example 229 from activator 991 (21 mg, 0.048 mmol) and linker 11 (20.2 mg, 0.048 mmol, 1.0 equiv). Brown solid (21.8 mg, yield 54%). 'H NMR (400 MHz, CD3OD) δ 7.64 - 7.53 (m, 2H), 7.52 - 7.33 (m, 6H), 7.23 (d, J= 8.7 Hz, 2H), 6.80 - 6.61 (m, 1H), 6.28 (dd, J= 16.8, 1.9 Hz, 1H), 6.22 - 6.13 (m, 1H), 6.07 (d, J= 3.2 Hz, 1H), 5.81 (dt, J= 10.6, 1.6 Hz, 1H), 4.37 (d, J= 25.5 Hz, 2H), 3.92 (t, J= 5.4 Hz, 2H), 3.87 - 3.74 (m, 5H), 3.70 - 3.52 (m, 8H), 3.47 (t, J= 5.5 Hz, 2H), 3.31 - 3.26 (m, 2H), 2.85 (t, J= 7.4 Hz, 2H), 2.50 - 2.39 (m, 5H). MS (ESI) [M+H]+= 836.3.

[0707] Example 240: JV-(15-(5-(4-acryloyl-2-oxopiperazin-l-yl)furan-2-yl)-13-oxo-3,6,9-trioxa-12- azapentadecyl)-5-((6-chloro-5-(l-methyl-1H-indol-5-yl)-17 / -benzo[t / |imidazol-2-yl)oxy)-2- methylbenzamide (XF137-92). Example 240 was synthesized following the standard procedure for preparing example 229 from activator 991 (21 mg, 0.048 mmol) and linker 12 (22 mg, 0.048 mmol, 1.0 equiv). Brown solid (22.9 mg, yield 61%).JH NMR (400 MHz, CD3OD) 5 7.59 (d, J = 7.6 Hz, 2H), 7.53 - 7.35 (m, 6H), 7.27 - 7.19 (m, 2H), 6.80 - 6.64 (m, 1H), 6.29 (dd, J= 16.7, 1.9 Hz, 1H), 6.22 (d, J= 3.3 Hz, 1H), 6.08 (d, J= 3.2 Hz, 1H), 5.81 (dd, J= 10.7, 1.9 Hz, 1H), 4.44 - 4.30 (m, 2H), 3.97 - 3.88 (m, 2H), 3.87 - 3.75 (m, 5H), 3.70 - 3.49 (m, 12H), 3.45 (t, J= 5.4 Hz, 2H), 3.30 (d, J= 5.5 Hz, 2H), 2.88 (t, J = 7.4 Hz, 2H), 2.51 - 2.44 (m, 5H). MS (ESI) [M+H]+=

[0708] 880.3.

[0709] Example 241: JV-(18-(5-(4-acryloyl-2-oxopiperazin-l-yl)furan-2-yl)-16-oxo-3,6,9,12- tetraoxa-15-azaoctadecyl)-5-((6-chloro-5-(l-methyl-1H-indol-5-yl)-1H-benzo[d ]imidazol-2- yl)oxy)-2-methylbenzamide (XF137-93). Example 241 was synthesized following the standard procedure for preparing example 229 from activator 991 (21 mg, 0.048 mmol) and linker 13 (24 mg, 0.048 mmol, 1.0 equiv). Brown solid (19.8 mg, yield 45%). 'H NMR (400 MHz, CD3OD) 8 7.58 (d, J= 5.8 Hz, 2H), 7.51 - 7.40 (m, 6H), 7.28 - 7.18 (m, 2H), 6.83 - 6.65 (m, 1H), 6.29 (dd, J = 16.8, 2.0 Hz, 1H), 6.22 (d, J= 3.2 Hz, 1H), 6.09 (d, J = 3.2 Hz, 1H), 5.81 (dd, J= 10.6, 1.9 Hz, 1H), 4.47 - 4.31 (m, 2H), 3.97 - 3.88 (m, 2H), 3.88 - 3.75 (m, 3H), 3.70 - 3.51 (m, 18H), 3.47 (t, J= 5.5 Hz, 2H), 3.38 - 3.36 (m, 2H), 2.89 (t, J= 7.4 Hz, 2H), 2.55 - 2.40 (m, 5H). MS (ESI) [M+H]+= 924.4.

[0710] Example 242: A-(21-(5-(4-acryloyl-2-oxopiperazin-l-yl)furan-2-yl)-19-oxo-3,6,9,12,15- pentaoxa-18-azahenicosyl)-5-((6-chloro-5-(l-methyl-1H-indol-5-yl)-177-benzo[d|imidazol-2- yl)oxy)-2-methylbenzamide (XF137-94). Example 242 was synthesized following the standard procedure for preparing example 229 from activator 991 (21 mg, 0.048 mmol) and linker 14 (27 mg, 0.048 mmol, 1.0 equiv). Brown solid (16.8 mg, yield 36%). 'H NMR (400 MHz, CD3OD) 8 7.59 (d, J= 5.8 Hz, 2H), 7.52 - 7.41 (m, 6H), 7.28 - 7.18 (m, 2H), 6.82 - 6.66 (m, 1H), 6.29 (dd, J = 16.7, 2.0 Hz, 1H), 6.23 (d, J = 3.2 Hz, 1H), 6.10 (d, J = 3.2 Hz, 1H), 5.82 (dd, J = 10.6, 1.9 Hz, 1H), 4.48 - 4.32 (m, 2H), 3.97 - 3.88 (m, 2H), 3.86 - 3.74 (m, 5H), 3.70 - 3.51 (m, 20H), 3.48 (t, J= 5.5 Hz, 2H), 3.39 - 3.36 (m, 2H), 2.90 (t, J= 7.4 Hz, 2H), 2.55 - 2.42 (m, 5H). MS (ESI) [M+H]+= 968.5.

[0711] Scheme 13. The syntheses of linker 15

[0712] Linker 15: (5-((6-chloro-5-(l-methyl-1H-indol-5-yl)-1H-benzo[d]imidazol-2-yl)oxy)-2- methylbenzoyl)glycine To a solution of activator 991 (50 mg, 0.12 mmol) in DMF (1 mL) were added tert-butyl (2-aminoethyl)carbamate (19.2 mg, 0.12 mmol, 1.0 equiv), l-ethyl-3-(3- dimethylaminopropyl)carbodiimide (EDCI, 34.3 mg, 0.18 mmol, 1.5 equiv), l-hydroxy-7- azabenzo-triazole (HO At, 24.5 mg, 0.18 mmol, 1.5 equiv), and N-methylmorpholine (NMM, 36.4 mg, 0.36 mmol, 3.0 equiv). After being stirred for overnight atrt, the resulting mixture was purified by prep-HPLC to afford crude product. The crude product was dissolved in DCM (1 mL) and TFA (1 mL). After being stirred at rt for 30 min, the resulting mixture was purified by prep-HPLC to afford compound titled compound as yellow solid (57 mg, yield 81%). MS (ESI) [M+H]+= 489.3. Linker 16-27 were synthesized following the same procedure for preparing linker 15. Linker 16: 3-(5-((6-chloro-5-(l-methyl-lH-indol-5-yl)-l / 7-benzo[J]imidazol-2-yl)oxy)-2- methylbenzamido)propanoic acid MS (ESI) [M+H]+= 503.2.

[0713] Linker 17: 4-(5-((6-chloro-5-(l-methyl-1H-indol-5-yl)-1H-benzo[d|imidazol-2-yl)oxy)-2- methylbenzamido)butanoic acid MS (ESI) [M+H]+=517.4.

[0714] Linker 18: 5-(5-((6-chloro-5-(l-methyl-1H-indol-5-yl)-1H-benzo[d]imidazol-2-yl)oxy)-2- methylbenzamido)pentanoic acid MS (ESI) [M+H]+= 531.3.

[0715] Linker 19: 6-(5-((6-chloro-5-(l-methyl-1H-indol-5-yl)-1H-benzod / |imidazol-2-yl)oxy)-2- methylbenzamido)hexanoic acid MS (ESI) [M+H]+= 545.3.

[0716] Example 110: (E)-N -(l-(4-(dimethylamino)but-2-enoyl)azetidin-3-yl)-5-methylthiophene-2- carboxamide (XS185-171). Example 110 was synthesized following similar procedure for preparing example 104. White solid, 15% yield1H NMR (400 MHz, DMSO-Jc,) 88.54 - 8.35 (m, 1H), 7.34 (d, J= 4.1 Hz, 4H), 7.29 - 7.20 (m, 1H), 6.67 - 6.49 (m, 2H), 4.81 (d, .7 = 21.1 Hz, 2H), 4.25 (s, 2H), 4.19 (d, J= 4.8 Hz, 2H), 3.91 - 3.82 (m, 2H), 3.82 - 3.69 (m, 2H), 2.36 (s, 3H).MS (ESI) [M+H]+=424.2

[0717] Linker 20: 7-(5-((6-chloro-5-(l-methyl-1H-indol-5-yl)- 1H-benzo[d|imidazol-2-yl)oxy)-2- methylbenzamido)heptanoic acid MS (ESI) [M+H]+= 559.4.

[0718] Linker 21 : 8-(5-((6-chloro-5-(l-methyl-1H-indol-5-yl)-1H-benzo[d / ]imidazol-2-yl)oxy)-2- methylbenzamido)octanoic acid MS (ESI) [M+H]+= 573.3.

[0719] Linker 22: 9-(5-((6-chloro-5-(l-methyl-1H-indol-5-yl)-l / / -benzo[d]imidazol-2-yl)oxy)-2- methylbenzamido)nonanoic acid MS (ESI) [M+H]+= 587.5.

[0720] Linker 23: 3-(2-(5-((6-chloro-5-(l-methyl-1H-indol-5-yl)-l / 7-benzo[<Z|iniidazol-2-yl)oxy)-2- methylbenzamido)ethoxy)propanoic acid MS (ESI) [M+H]+= 548.6.

[0721] Linker 24: 3-(2-(2-(5-((6-chloro-5-(l-methyl-1H-indol-5-yl)-1H-benzo[d / |iniidazol-2-yl)oxy)- 2-methylbenzamido)ethoxy)ethoxy)propanoic acid MS (ESI) [M+H]+= 591.4.

[0722] Linker 25: l-(5-((6-chloro-5-(l-methyl-l / / -indol-5-yl)-177-benzo[d |iniidazol-2-yl)oxy)-2- methylphenyl)-l-oxo-5,8,ll-trioxa-2-azatetradecan-14-oic acid MS (ESI) [M+H]+= 635.4.

[0723] Linker 26: l-(5-((6-chloro-5-(l-methyl-l / / -indol-5-yl)-177-benzo[d|iniidazol-2-yl)oxy)-2- methylphenyl)-l-oxo-5,8,ll,14-tetraoxa-2-azaheptadecan-17-oic acid MS (ESI) [M+H]+= 679.4.

[0724] Linker 27: l-(5-((6-chIoro-5-(1-methyl-1H-indol-5-yl)-1H-benzo[d]imidazol-2-yl)oxy)-2- methylphenyl)-l-oxo-5,8,ll,14,17-pentaoxa-2-azaicosan-20-oic acid MS (ESI) [M+H]+=

[0725] 723.3.

[0726] Scheme 14. The syntheses of linker 28 activator 991 linker 28

[0727] Linker 28 : JV-(2-aminoethyl)-5-((6-chloro-5-(l-methyl-1H-indol-5-yl)-lH-benzo[d ]imidazol- 2-yl)oxy)-2-methylbenzamide (XH168-164). To a solution of activator 991 (50 mg, 0.12 mmol) in DMF (1 mL) were added tert-butyl (2-aminoethyl)carbamate (19.2 mg, 0.12 mmol, 1.0 equiv), l-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI, 34.3 mg, 0.18 mmol, 1.5 equiv), 1- hydroxy-7-azabenzo-triazole (HO At, 24.5 mg, 0.18 mmol, 1.5 equiv), and N-methylmorpholine (NMM, 36.4 mg, 0.36 mmol, 3.0 equiv). After being stirred for overnight at rt, the resulting mixture was purified by preparative HPLC ( 10%- 100% acetonitrile / 0.1% TFA in H2O) to afford crude product. The crude product was dissolved in DCM (1 mL) and TFA (1 mL). After being stirred at rt for 30 min, the resulting mixture was purified by preparative HPLC (10%-100% acetonitrile / 0.1% TFA in H2O) to afford compound titled compound as yellow solid (57 mg, yield 81%). MS (ESI) m / z 474.4 [M+H]+.

[0728] Linker 29-41 was synthesized following the same procedure for preparing linker 28.

[0729] Linker 29: N -(3-aminopropyl)-5-((6-chloro-5-(l-methyl-1H-indol-5-yl )-1H- benzo[* / ]imidazol-2-yl)oxy)-2-methylbenzamide (XH168-165). Pink solid, 52 mg, 61% yield.

[0730] MS (ESI) [M+H]+= 488.3.

[0731] Linker 33: N-(7-aminoheptyl)-5-((6-chloro-5-(l-methyl-1H-indol-5-yl)-1H- benzo[d |imidazol-2-yl)oxy)-2-methylbenzamide (XH168-169). Yellow solid, 73 mg, 79% yield.

[0732] MS (ESI) [M+H]+= 544.5. Linker 34: N -(8-aminooctyl)-5-((6-chloro-5-(1-methyl-1H-indol-5-yl)-1H-benzo[d]imidazol- 2-yl)oxy)-2-methylbenzamide (XH168-170). Yellow solid, 69 mg, 73% yield. MS (ESI) [M+H]+= 558.4.

[0733] Linker 36: JV-(10-aminodecyl)-5-((6-chloro-5-(l-methyl-1H-indol-5-yl)-EH- benzo[rf|imidazol-2-yl)oxy)-2-methylbenzamide (XH168-172) Yellow solid, 75 mg, 87% yield. MS (ESI) [M+H]+= 586.5.

[0734] Linker 37: N -(2-(2-aminoethoxy)ethyl)-5-((6-chloro-5-(l-methyl-1H-indol-5-yl)-1H- benzo|d]imidazol-2-yl)oxy)-2-methylbenzamide (XH168-173). Yellow solid, 65 mg, 73% yield. MS (ESI) [M+H]+= 518.1. Linker 38: N -(2-(2-(2-aminoethoxy)ethoxy)ethyl)-5-((6-chloro-5-(l-methyl-1H-indol-5-yl)-1H-benzo[d]imidazol-2-yl)oxy)-2-methylbenzamide (XH168-174). Yellow solid, 79 mg, 81% yield. MS (ESI) [M+H]+= 562.5.

[0735] Linker 41: N-(17-amino-3,6,9,12,15-pentaoxaheptadecyl)-5-((6-chloro-5-(l-methyl-l1H- indol-5-yl)-1H-benzo[d]imidazol-2-yl)oxy)-2-niethylbenzaniide (XH168-177). Yellow solid, 89 mg, 81% yield. MS (ESI) [M+Hf = 694.5.

[0736] Scheme 15. Synthesis of intermediate 13.

[0737]

[0738] Intermediate 13

[0739] (E)-3-(5-(4-(4-(dimethylamino)but-2-enoyl)-2-oxopiperazin-l-yl)thiophen-2-yl)propanoic acid (Intermediate 13) methyl 3-(5-bromothiophen-2-yl)acrylate (246 mg, 1.0 mmol, 1.0 eq), tert-butyl 3-oxopiperazine-l -carboxylate (300 mg, 1.5 mmol, 1.5 eq), Cui (85 mg, 0.5 mmol), dimethylethane-l,2-diamine (88 mg, 1.0 mmol, 1.0 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq) were added Toluene ( 5 mL). The mixture was stirred at 110°C under N2 for 6 h. The mixture was purified by ISCO. The obtained product was then dissolved in MeOH (5 mL) followed by Pd / C (60 mg) ed. The mixture was stirred at room temperature under 1 atm H2 for 1 h then filtered to yield intermediate 12 as a white solid (200 mg, 55%). 'H NMR (400 MHz, CDCI3) 8 6.62 (s, 1H), 6.48 (s, 1H), 4.28 (s, 2H), 3.81 (s, 4H), 3.68 (s, 3H), 3.08 (t, J= 7.7 Hz, 2H), 2.68 (d, J= 7.7 Hz, 2H), 1.48 (s, 9H). MS (ESI) [M+H]+= 369.2.

[0740] Intermediate 13 (184 mg, 0.5 mmol, 1.0 eq) was dissolved in MeOH (2 mL) and HC1 (4M in dioxane, 2 mL, 8 mmol), the mixture was stirred at room temperature for 3h, then removed all the volatiles. The resulted residue (71mg, 0.55 mmol, 1.1 eq), HATU (285 mg, 0.75 mmol, 1.5 eq), DIPEA (194 mg, 1.5 mmol, 3.0 eq) were stirred in DCM (5 mL) at room temperature for 30 min. Then removed all the volatiles, and the mixture was purified via reverse-ISCO. The purified compound (120 mg, 0.33 mmol, 1.0 eq) was dissolved in MeOH (2 mL). LiOH aq. (2M, 0.5 mL, 1 mmol) was added dropwise at 0 °C. The mixture was stirred at 0 °C for 20 min, then was purified via reverse-ISCO to yield title compound as a brown solid (100 mg, 83%).1H NMR (400 MHz, CD3OD) δ 7.08 - 6.64 (m, 4H), 4.13 - 3.89 (m, 6H), 3.15 - 3.04 (m, 2H), 2.98 - 2.87 (m, 8H), 2.73 - 2.62 (m, 2H). MS (ESI) [M+H]+= 366.3.

[0741] Scheme 16. Synthesis of intermediate 15

[0742] (£')-l-(5-(2-aminoethyl)thiophen-2-yl)-4-(4-(dimethylamino)but-2-enoyl)piperazin-2-one (Intermediate 15) tert-butyl (2-(5-bromothiophen-2-yl)ethyl)carbamate (306 mg, 1.0 mmol, 1.0 eq), benzyl 3-oxopiperazine-l-carboxylate (350 mg, 1.5 mmol, 1.5 eq), Cui (85 mg, 0.5 mmol), N1-dimethylethane-l,2-diamine (88 mg, 1.0 mmol, 1.0 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq) were added Toluene (5 mL). The mixture was stirred at 110 °C under N2 for 6 h. The mixture was purified ISCO to yield intermediate 14 as a white solid (278 mg, 61%). 'H NMR (400 MHz, CD3OD) 5 7.47 - 7.32 (m, 5H), 6.69 (s, 2H), 5.20 (s, 2H), 4.33 (s, 2H), 3.91 (s, 4H), 3.33 - 3.25 (m, 2H), 2.94 - 2.87 (m, 2H), 1.45 (s, 9H). MS (ESI) [M+H]+= 460.4.

[0743] Intermediate 14 (459 mg, 1.0 mmol, 1.0 eq), Pd / C (120 mg), Pd(OH)2 (120 mg) were added to MeOH (5 mL), the mixture was stirred under 10 atm H2 at 60 °C for 2 h. The mixture was filtered and the product was used without further purification. MS (ESI) [M+H]+= 326.3. The obtained crude product (160 mg, 0.5 mmol, 1.0 eq), (E)-4-(dimethylamino)but-2-enoic acid (71mg, 0.55 mmol, 1.1 eq), HATU (285 mg, 0.75 mmol, 1.5 eq), DIPEA (194 mg, 1.5 mmol, 3.0 eq) were stirred in DCM (5 mL) at room temperature for 30 min. Then removed all the volatiles, and the mixture was purified via reverse-ISCO. The obtained product was stirred with DCM / TFA (1 mL / lmL) for 20 min. Then removed all the volatiles, and the mixture was purified via reverse- ISCO to yield title compound as a white solid (150 mg, 98%). 'H NMR (400 MHz, CD3OD) 8 7.17 - 6.74 (m, 4H), 4.19 - 3.93 (m, 6H), 3.26 - 3.19 (m, 2H), 3.18 - 3.11 (m, 2H), 3.01 - 2.90 (m, 8H). MS (ESI) [M+H]+= 337.2.

[0744] Scheme 17. The syntheses of example 243

[0745] Example 243

[0746] Example 243: (E)-5-((6-chloro-5-(l-methyl-1H-indol-5-yl)-1H-benzo[d|imidazol-2-yl)oxy)-A- (7-((2-(5-(4-(4-(dimethylamino)but-2-enoyl)-2-oxopiperazin-l-yl)thiophen-2- yl)ethyl)amino)-7-oxoheptyl)-2-methylbenzamide (QC179-047) Linker 20 (12 mg, 0.025 mmol, 1.0 eq), intermediate 15 (10 mg, 0.028 mmol, 1.1 eq), EDC HC1 (7 mg, 0.038 mmol, 1.5 eq), HOAt (5 mg, 0.038 mmol, 1.5 eq), NMM (8 mg, 0.075 mmol, 3.0 eq) were stirred in 2 mL DMF at room temperature for 2 h. Then the mixture was purified via Prep-HPLC to yield titled compound as a white solid (8 mg, 46% yield).1H NMR (400 MHz, CD3OD) 8 7.58 (s, 1H), 7.51 (s, 1H), 7.46 - 7.32 (m, 5H), 7.27 - 7.21 (m, 2H), 6.94 (dd, J= 32.4, 15.1 Hz, 1H), 6.77 (br, 1H), 6.70 - 6.64 (m, 2H), 6.49 (s, 1H), 4.50 (s, 1H), 4.43 (s, 1H), 4.04 (s, 2H), 3.95 (s, 3H), 3.87 (s, 4H), 3.44 - 3.36 (m, 4H), 2.92 (s, 8H), 2.46 (s, 3H), 2.24 - 2.13 (m, 2H), 1.64 (s, 4H), 1.47 - 1.30 (m, 4H). MS (ESI) [M+H]+= 877.5.

[0747] Example 244: (E)-5-((6-chloro-5-(l-methyl-1H / -indol-5-yl)-1H-benzo[d|imidazol-2-yl)oxy)-A- (8-((2-(5-(4-(4-(dimethylamino)but-2-enoyl)-2-oxopiperazin-l-yl)thiophen-2- yl)ethyl)amino)-8-oxooctyl)-2-methylbenzamide (QC179-048) Example 244 was synthesized following the same procedure for preparing example 243 from linker 21. White solid, 9 mg, 50% yield.1H NMR (400 MHz, CD3OD) δ 7.67 - 7.11 (m, 10H), 7.02 - 6.85 (m, 1H), 6.78 (br, 1H), 6.68 - 6.56 (m, 2H), 4.52 - 4.39 (m, 2H), 4.13 - 3.86 (m, 9H), 3.40 (s, 4H), 2.91 (s, 8H), 2.45 (s, 3H), 2.15 (s, 2H), 1.61 (s, 4H), 1.46 - 1.30 (s, 6H). MS (ESI) [M+H]+= 891.5.

[0748] Example 245: (E)-5-((6-chloro-5-(l-methyl-1Hindol-5-yl)-1Hbenzo[d|imidazol-2-yl)oxy)-N- (9-((2-(5-(4-(4-(dimethylamino)but-2-enoyl)-2-oxopiperazin-l-yl)thiophen-2- yl)ethyl)amino)-9-oxononyl)-2-methylbenzamide (QC179-049) Example 245 was synthesized following the same procedure for preparing example 243 from linker 22. White solid, 9 mg, 45% yield.1H NMR (400 MHz, CD3OD) 8 7.58 (s, 1H), 7.51 (s, 1H), 7.44 - 7.36 (m, 5H), 7.29 - 7.20 (m, 2H), 6.95 (dd, J= 33.6, 15.2 Hz, 1H), 6.77 (br, 1H), 6.67 (s, 2H), 6.49 (s, 1H), 4.50 (s, 1H), 4.44 (s, 1H), 4.04 (s, 2H), 3.98 - 3.92 (m, 3H), 3.91 - 3.95 (m, 4H), 3.38 (s, 4H), 2.92 (s, 8H), 2.46 (s, 3H), 2.20 - 2.11 (m, 2H), 1.68 - 1.54 (m, 4H), 1.48 -1.24 (m, 8H). MS (ESI) [M+H]+= 905.6.

[0749] Example 246: (E)-l-(5-((6-chloro-5-(l-methyl-1H-indol-5-yl)-l / / -benzo[d|imidazol-2- yl)oxy)-2-methylbenzamido)-N -(2-(5-(4-(4-(dimethylamino)but-2-enoyl)-2-oxopiperazin-l- yl)thiophen-2-yl)ethyl)-3,6,9,12,15-pentaoxaoctadecan-18-amide (QC179-050) Example 246was synthesized following the same procedure for preparing example 243 from linker 27. White solid, 8 mg, 40% yield.1H NMR (400 MHz, CD3OD) 8 7.58 (s, 1H), 7.52 (s, 1H), 7.45 - 7.37 (m, 5H), 7.27 - 7.21 (m, 2H), 7.00 - 6.87 (m, 1H), 6.77 (s, 1H), 6.69 (s, 1H), 6.65 (s, 1H), 6.49 (s, 1H), 4.49 (s, 1H), 4.43 (s, 1H), 4.03 (s, 2H), 3.98 - 3.93 (m, 3H), 3.87 (s, 4H), 3.66 - 3.50 (m, 26H), 2.92 (s, 8H), 2.47 (s, 3H). MS (ESI) [M+H]+= 1141.6.

[0750] Scheme 18. The syntheses of example 247

[0751] Example 247

[0752] Example 247: (£)-A-(2-((5-((6-chloro-5-(l-methyl-l / / -indol-5-yl)-l / / -benzo[J|imidazol-2- yl)oxy)-2-methylbenzoyl)tetramethyl-l7-azaneyl)ethyl)-3-(5-(4-(4-(dimethylamino)but-2- enoyl)-2-oxopiperazin-l-yl)thiophen-2-yl)propenamide (QC179-051) Linker 32 (14 mg, 0.020 mmol, 1.0 eq), intermediate 13 (11 mg, 0.022 mmol, 1.1 eq), EDC HC1 (6 mg, 0.030 mmol, 1.5 eq), HOAt (4 mg, 0.030 mmol, 1.5 eq), NMM (6 mg, 0.060 mmol, 3.0 eq) were stirred in 1 mL DMF at room temperature for 2 h. Then the mixture was purified via Prep-HPLC to yield titled compound as a white solid (8 mg, 40% yield). 'H NMR (400 MHz, CD3OD) δ 7.57 (s, 1H), 7.51 (s, 1H), 7.45 - 7.36 (m, 5H), 7.27 - 7.21 (m, 2H), 6.93 (dd, J= 31.9, 14.9 Hz, 1H), 6.76 (s, 1H), 6.63 (s, 2H), 6.49 (s, 1H), 4.49 (s, 1H), 4.43 (s, 1H), 4.03 (s, 2H), 3.99 - 3.93 (m, 3H), 3.87 (s, 4H), 3.15 (s, 2H), 3.08 - 3.00 (m, 2H), 2.92 (s, 6H), 2.54 - 2.44 (m, 5H), 1.61 (s, 2H), 1.53 - 1.28 (m, 8H). MS (ESI) [M+H]+= 877.4.

[0753] Example 248: (E)-A-(2-((5-((6-chloro-5-(l-methyl-17 / -indol-5-yl)-1H-benzo[cZ|imidazol-2- yl)oxy)-2-methylbenzoyl)pentamethyl-l8-azaneyl)ethyl)-3-(5-(4-(4-(dimethylamino)but-2- enoyl)-2-oxopiperazin-l-yl)thiophen-2-yl)propenamide (QC179-052) Example 248 was synthesized following the same procedure for preparing example 247 from linker 33. White solid 7 mg, 43% yield.1H NMR (400 MHz, CD3OD) 8 7.58 (s, 1H), 7.51 (s, 1H), 7.43 - 7.36 (m, 5H), 7.27 - 7.21 (m, 2H), 7.01 - 7.88 (m, 1H), 6.76 (s, 1H), 6.64 (s, 2H), 6.49 (s, 1H), 4.51 - 4.37 (m, 2H), 4.08 - 4.02 (m, 2H), 3.98 - 3.93 (m, 3H), 3.87 (s, 4H), 3.18 - 3.11 (m, 2H), 3.07 -3.01 (m, 2H), 2.92 (s, 6H), 2.54 - 2.43 (m, 5H), 1.62 - 1.32 (m, 12H). MS (ESI) [M+H]+= 891.4.

[0754] Example 249: (E)-N-(2-((5-((6-chloro-5-(l-methyl-1H-indol-5-yl)-1H-benzo[d|imidazol-2- yl)oxy)-2-methylbenzoyl)hexamethyl-l9-azaneyl)ethyl)-3-(5-(4-(4-(dimethylamino)but-2- enoyl)-2-oxopiperazin-l-yl)thiophen-2-yl)propenamide (QC179-053) Example 249 was synthesized following the same procedure for preparing example 247 from linker 34. White solid, 10 mg, 48% yield. 'H NMR (400 MHz, CD3OD) δ 7.58 (s, 1H), 7.51 (s, 1H), 7.44 - 7.35 (m, 5H), 7.27 -7.21 (m, 2H), 6.95 (dd, J = 35.2, 15.7 Hz, 1H), 6.77 (s, 1H), 6.67 - 6.47 (m, 3H), 4.51 - 4.38 (m, 2H), 4.05 - 3.85 (m, 9H), 3.17 - 3.05 (m, 4H), 2.92 (s, 6H), 2.52 - 2.44 (m, 5H), 1.62 (s, 2H), 1.43 - 1.23 (m, 12H). MS (ESI) [M+H]+ = = 905.4.

[0755] Example 250: (E)-N-(2-((5-((6-chloro-5-(l-methyl-1H-indol-5-yl)-1H-benzo[d|imidazol-2- yl)oxy)-2-methylbenzoyl)heptamethyl-l10-azaneyl)ethyl)-3-(5-(4-(4-(dimethylamino)but-2- enoyl)-2-oxopiperazin-l-yl)thiophen-2-yl)propenamide (QC179-054) Example 250 was synthesized following the same procedure for preparing example 247 from linker 35. White solid 9 mg, 50% yield. 'HNMR (400 MHz, CD3OD) δ 7.58 - 7.14 (m, 10H), 7.02 - 6.88 (m, 1H), 6.77 (s, 1H), 6.66 - 6.49 (m, 2H), 4.41 (s, 2H), 4.03 - 3.71 (m, 9H), 3.13 - 2.99 (m, 4H), 2.91 (s, 6H), 2.53 - 2.44 (m, 5H), 1.60 (s, 2H), 1.41 - 1.23 (m, 14H). MS (ESI) [M+H]+= 919.3.

[0756] Example 251: (E )-N-(2-((5-((6-chloro-5-(1-methyl-1H-indol-5-yl)-1H-benzo|d|imidazol-2- yl)oxy)-2-methylbenzoyl)octamethyl-l11-azaneyl)ethyl)-3-(5-(4-(4-(dimethylamino)but-2- enoyl)-2-oxopiperazin-l-yl)thiophen-2-yl)propenamide (QC179-055) Example 251 was synthesized following the same procedure for preparing example 247 from linker 36. White solid 7 mg, 40% yield. ’H NMR (400 MHz, CD3OD) 8 7.58 (s, 1H), 7.52 (s, 1H), 7.45 - 7.34 (m, 5H), 7.28 - 7.21 (m, 2H), 6.94 (dd, J= 34.8, 15.2 Hz, 1H), 6.78 (s, 1H), 6.67 - 6.60 (m, 2H), 6.49 (s, 1H), 4.49 (s, 1H), 4.43 (s, 1H), 4.04 (s, 2H), 3.99 - 3.85 (m, 7H), 3.16 - 3.03 (m, 4H), 2.92 (s, 6H), 2.55 - 2.42 (m, 5H), 1.64 (s, 2H), 1.47 - 1.22 (m, 16H). MS (ESI) [M+H]+= 933.5.

[0757] Procedures for the synthesis of cGAS based bivalent compounds

[0758] Scheme 19. The syntheses of linker 42 Linker 42: 2-amino-A-(2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-lH- pyrido[4,3-b]indoI-9-yl)-1H-pyrazol-l-yl)ethyl)acetamide To a solution of 3-(4, 4,5,5- tetramethyl-l,3,2-dioxaborolan-2-yl)-177-pyrazole (7.3 g, 37.6 mmol, 1.0 eq) in MeCN (30ml) was added K2CO3 (15.6 g, 112.9 mmol, 3.0 eq) and tert-butyl (2-bromoethyl)carbamate (10.1 g, 45.1 mmol, 1.2 eq). The mixture was stirred at 80 °C for 6h. After that, water was added and the mixture was extracted with ethyl acetate. The organic layer was collected, and excess solvent was removed to get intermediate 16 and used for the next step directly without further purification.

[0759] To a solution of intermediate 17 (1.05 g, 2.5 mmol, 1 eq) (prepared following previous reported lit.) (Lama et al., 2019) and intermediate 16 (3.2 g, 9.5 mmol, 3.8 eq) in dioxane (40 ml) were added KOAc (981 mg, 10.0 mmol, 4.0 eq) and Pd(dppf)Cl2(408 mg, 0.5 mmol, 0.2 eq). The mixture was stirred at 100 °C for 6 h under N2 followed concentrated under reduced pressure. The resulted residue was purified by column chromatography (SiCL, DCM / MeOH = 10: 1) to give intermediate 18 as a brown oil (894 mg, 65% yield).1H NMR (400 MHz, DMSO-d6) δ 7.88 (s, 1H), 7.32 (d, J = 9.7 Hz, 1H), 6.70 (dd, J = 27.9, 2.3 Hz, 1H), 4.82 (d, J = 61.8 Hz, 2H), 4.68 - 4.36 (m, 4H), 3.76 (dt, J= 45.2, 5.9 Hz, 2H), 2.89 (dt, J= 46.7, 5.8 Hz, 2H), 2.07 (d, J= 1.9 Hz, 5H), 1.11 (s, 9H).

[0760] To a solution of intermediate 18 (420 mg, 0.76 mmol, 1 eq) in MeOH / H2O (10:1, 4.4 ml) was added LiOH.H2O (91 mg, 2.3 mmol, 3 eq). The mixture was stirred at rt for 10 min followed by concentrated and diluted with H2O. After that, the pH value of the solution was adjusted to 6 with HC1 aq. solution (IM) followed by concentration. The residue was re-dissolved in DCM / TFA (2: 1, 7.5 mL). After stirring at rt for Ih, excess solvent was removed and the residue was purified by prep-HPLC to give intermediate 19 as a yellow solid, (178.6 mg, 45% yield).1H NMR (400 MHz, CD3OD) 8 7.77 (d, J= 13.4 Hz, IH), 7.23 (dd, J = 9.9, 3.0 Hz, IH), 6.56 (d, J= 9.3 Hz, IH), 4.73 - 4.47 (m, 4H), 4.40 - 4.13 (m, 2H), 3.97 - 3.46 (m, 4H), 3.07 - 2.81 (m, 2H).

[0761] To a solution of succinic acid (35.4mg, 0.3 mmol, 2.0 eq) in DMF (0.75 mL) and DCM (18 mL) was added NMM (75.9mg, 0.75 mmol, 5.0 eq), intermediate 18 (78.3mg, 0.15 mmol, 1.0 eq), HO At (24.5mg, 0.18 mmol, 1.2 eq), and EDCI (34.5mg, 0.18 mmol, 12eq) at 0°C. The resulted reaction solution was stirred at 0 °C for 6 h before being stirred at room temperature overnight followed by concentrated. The resulted residue was purified by reverse-phase chromatography to yield the titled product (38.1 mg, 50% yield). ’H NMR (400 MHz, CD3OD) 87.78 - 7.65 (m, IH), 7.21 (d, J= 18.1 Hz, IH), 6.49 (dd, J= 23.8, 2.3 Hz, IH), 4.55 (d, J= 7.0 Hz, 2H), 4.42 - 4.16 (m, 4H), 3.99 - 3.61 (m, 4H), 2.94 (dt, J = 22.2, 5.8 Hz, 2H), 2.59 (t, J = 6.8 Hz, 2H), 2.46 (dt, J = 10.8, 6.7 Hz, 2H).

[0762] Linker 43 - 55 were synthesized following the same procedure for preparing linker 42. J= 25.4, 2.3 Hz, 1H), 4.65 - 4.17 (m, 6H), 3.79 - 3.56 (m, 4H), 3.02 - 2.74 (m, 2H), 2.40 - 2.03 (m, 4H), 1.40 (s, 6H).

[0763] Linker 48: 10-((2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-1H-pyrido[4,3- b]indol-9-yl)-l / Z-pyrazol-l-yl)ethyl)amino)-10-oxodecanoic acid. Brown solid, yield 60%. I I NMR (400 MHz, CD3OD) 6 7.65 (dd, J= 20.7, 2.3 Hz, 1H), 7.20 (d, J= 14.7 Hz, 1H), 6.48 (dd, J= 24.8, 2.3 Hz, 1H), 4.65 - 4.17 (m, 6H), 3.97 - 3.56 (m, 4H), 2.92 (d, J= 27.6 Hz, 2H), 2.18 (t, J= 59.4 Hz, 4H), 1.67 - 1.04 (m, 12H).

[0764] Linker 49: 1 l-((2-(3-(6.7-dichloro-2-(2-hydroxya cetyl)-2.3.4.5-tetrahydro-l / / -pyrido|4.3- / >|indol-9-yl)-l / / -pyrazol-l-yl)ethyl)amino)- 11-oxoundecanoic acid Brown solid, yield 47%. *H NMR (400 MHz, CD3OD) 8 7.65 (dd, J = 20.6, 2.3 Hz, 1H), 7.21 (d, J = 13.3 Hz, 1H), 6.49 (dd, J= 24.3, 2.3 Hz, 1H), 4.70 - 4.10 (m, 6H), 3.97 - 3.59 (m, 1H), 3.05 - 2.80 (m, 2H), 2.30 - 2.03 (m, 4H), 1.64 - 0.92 (m, 14H).

[0765] Linker 50: 12-((2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-lH-pyrido[4,3- / >Jindol-9-yl)-l / / -pyrazol-l-yl)ethyl)amino)-12-oxododecanoic acid. Brown solid, yield 45%.JH NMR (400 MHz, CD3OD) 8 7.65 (dd, J = 21.4, 2.0 Hz, 1H), 7.21 (d, J = 13.7 Hz, 1H), 6.48 (dd, J= 25.4, 2.2 Hz, 1H), 4.70 - 4.18 (m, 6H), 4.00 - 3.56 (m, 4H), 3.09 - 2.81 (m, 2H), 2.38 - 2.02 (m, 4H), 1.64 - 0.93 (m, 16H).

[0766] Linker 51: 3-(3-((2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-1H-pyrido[4,3- b]indol-9-yl)-1H-pyrazol-l-yl)ethyl)amino)-3-oxopropoxy)propanoic acid. Brown solid, yield 46%.1H NMR (400 MHz, CD3OD) 8 7.69 (dd, J= 15.3, 2.3 Hz, 1H), 7.20 (d, J= 17.8 Hz, 1H), 6.49 (dd, J = 23.4, 2.3 Hz, 1H), 4.62 - 4.17 (m, 6H), 3.97 - 3.56 (m, 8H), 3.02 - 2.82 (tn, 2H), 2.57 - 2.31 (m, 4H).

[0767] Linker 52: 3-(2-(3-((2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-1H- pyrido[4,3-6]indol-9-yl)-l / / -pyrazol-l-yl)ethyl)amino)-3-oxopropoxy)ethoxy)propanoic acid Brown solid, yield 49%. 'H NMR (400 MHz, CD3OD) 8 7.70 (dd, J= 17.0, 2.2 Hz, 1H), 7.20 (d, J= 16.8 Hz, 1H), 6.48 (dd, J = 24.2, 2.3 Hz, 1H), 4.62 - 4.15 (m, 6H), 3.97 - 3.42 (m, 12H), 3.01 - 2.82 (m, 2H), 2.54 - 2.32 (m, 4H).

[0768] Linker 53: l-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-1H-pyrido[4,3-i]indol- 9-yl)-1H-pyrazol-l-yl)-4-oxo-7,10,13-trioxa-3-azahexadecan-16-oic acid. Brown solid, yield .40 (m, Linker 54: l-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-lH-pyrido[4,3-b]indol- 9-yl)-1H-pyrazol-l-yl)-4-oxo-7,10,13,16-tetraoxa-3-azanonadecan-19-oic acid. Brown solid, yield 53 %. ’H NMR (400 MHz, CD3OD) 5 7.80 - 7.66 (m, 1H), 7.23 (d, J= 17.3 Hz, 1H), 6.52 (dd, J= 24.5, 2.3 Hz, 1H), 4.63 - 4.17 (m, 6H), 3.79 - 3.47 (m, 20H), 3.01 - 2.86 (m, 2H), 2.63 - 2.28 (m, 4H).

[0769] Linker 55: l-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-lH-pyrido[4,3- / >]indol- 9-yl)-lJ / -pyrazol-l-yl)-4-oxo-7,10,13,16,19-pentaoxa-3-azadocosan-22-oic acid. Brown solid, yield 47 %. 'H NMR (400 MHz, CD3OD) δ 7.79 - 7.49 (m, 1H), 7.11 (d, J = 17.3 Hz, 1H), 6.40 (dd, J= 24.3, 2.3 Hz, 1H), 4.54 - 4.10 (m, 6H), 3.73 - 3.38 (m, 24H), 2.92 - 2.76 (m, 2H), 2.51 - 2.14 (m, 4H).

[0770] Scheme 20. The syntheses of linker 56

[0771] Intermediate 19 Linker 56

[0772] Linker 56: 2-amino-N-(2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-1H- pyrido[4,3-b]indoI-9-yl)-l / 7-pyrazol-l-yl)ethyl)acetamide To a solution of intermediate 19 (65 mg, 0. 12 mmol) in DMF (0.5 mL) were added A-Boc glycine (19.2 mg, 0.12 mmol, 1.0 equiv), 1- ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI, 34.3 mg, 0.18 mmol, 1.5 equiv), 1- hydroxy-7-azabenzo-triazole (HOAt, 24.5 mg, 0.18 mmol, 1.5 equiv), and A-methylmorpholine (NMM, 36.4 mg, 0.36 mmol, 3.0 equiv). After being stirred overnight at rt, the resulting mixture was purified by prep-HPLC to afford crude product. The crude product was dissolved in DCM / TFA (1 :1, 2 mL) followed by stirred at rt for 30 min. The resulting mixture was purified by prep-HPLC to afford title compound as yellow solid (44 mg, 63% yield). 'H NMR (400 MHz, CD3OD) 8 7.78 - 7.65 (m, 1H), 7.21 (d, J= 17.2 Hz, 1H), 6.51 (dd, J= 26.0, 2.0 Hz, 1H), 4.60 - 4.50 (m, 2H), 4.44 - 4.30 (m, 4H), 3.00 - 2.87 (m, 6H), 7.78 - 7.65 (m, 2H).

[0773] Linker 57 - 70 were synthesized following the same procedure for preparing linker 56.

[0774] Linker 58: 4-amino-JV-(2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-1H- pyrido[4,3-b]indol-9-yl)-l / 7-pyrazol-l-yl)ethyl)butanamide. Brown solid, yield 41%.1HNMR (400 MHz, CD3OD) 87.67 (dd, J= 18.2, 2.3 Hz, 1H), 7.20 (d, J= 17.4 Hz, 1H), 6.49 (dd, J= 28.0, 2.3 Hz, 1H), 4.61 - 4.30 (m, 6H), 3.81 - 3.58 (m, 4H), 3.03 - 2.77 (m, 4H), 2.35 (t, J= 7.1 Hz, 2H), 2.01 - 1.80 (m, 2H).

[0775] Linker 59: 5-amino-N-(2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-1H- pyrido[4,3-6]indol-9-yl)-1H-pyrazol-l-yl)ethyl)pentanamide. Brown solid, yield 44%.JH NMR (400 MHz, CD3OD) 5 7.78 - 7.58 (m, 1H), 7.21 (d, J= 14.0 Hz, 1H), 6.67 - 6.29 (m, 1H), 4.65 - 4.18 (m, 6H), 4.02 - 3.61 (m, 4H), 4.02 - 3.62 (m, 4H), 2.29 - 2.21 (m, 2H), 1.72 - 1.59

[0776] Linker 60: 6-amino-N -(2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-1H- pyrido[4,3-6]indol-9-yl)-LH-pyrazol-l-yl)ethyl)hexanamide. Brown solid, yield 47%. ’l l NMR (400 MHz, CD3OD) 8 7.68 (dd, J = 20.8, 2.3 Hz, 1H), 7.20 (d, J = 14.7 Hz, 1H), 6.50 (dd, J = 24.4, 2.3 Hz, 1H), 4.65 - 4.30 (m, 6H), 3.75 - 3.66 (m, 4H), 2.98 - 2.84 (m, 4H), 2.30 - 2.02 (m, 2H), 1.65 - 1.53 (m, 4H), 1.44 - 1.24 (m, 2H).

[0777] Linker 61: 7-amino-N -(2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-1H- pyrido[4,3-b]indol-9-yl)-1H-pyrazol-l-yl)ethyl)heptanamide. Brown solid, yield 43%.1H NMR (400 MHz, CD3OD) 8 7.70 - 7.60 (m, 1H), 7.20 (d, J= 14.3 Hz, 1H), 6.50 (dd, J= 23.5, 2.3 Hz, 1H), 4.70 - 4.29 (m, 6H), 3.82 - 3.60 (m, 4H), 3.04 - 2.73 (m, 4H), 2.29 - 2.06 (m, 2H), 1.78 Linker 64: 10-amino- / V-(2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro1H - pyrido[4,3-^]indoI-9-yl)-l / 7-pyrazol-l-yl)ethyl)decanamide. Brown solid, yield 49%.1HNMR (400 MHz, CD3OD) 87.66 (dd, J= 21.3, 2.3 Hz, 1H), 7.21 (d, J= 12.8 Hz, 1H), 6.49 (dd, J= 24.3,

[0778] 2.3 Hz, 1H), 4.56 - 4.16 (m, 6H), 3.88 - 3.49 (m, 4H), 2.81 (dq, J= 28.2, 7.7, 6.7 Hz, 4H), 2.05 (dt, J= 16.3, 7.5 Hz, OH), 1.60 - 1.03 (m, 14H).

[0779] Linker 66: 3-(2-aminoethoxy)-JV-(2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro- l / / -pyrido|4,3-B)|indol-9-yl)- l / / -pyrazol-l-yl)ethyl)propenainide. Brown solid, yield 42%.1H NMR (400 MHz, CD3OD) 8 7.69 (dd, J = 17.1, 2.3 Hz, 1H), 7.21 (d, J= 16.7 Hz, 1H), 6.51 (dd, .7= 26.1, 2.3 Hz, 1H), 4.62 - 4.29 (m, 6H), 3.83 - 3.55 (m, 8H), 3.18 - 2.85 (m, 4H), 2.61 - 2.23 (m, 2H).

[0780] Linker 67: 3-(2-(2-aminoethoxy)ethoxy)-N -(2-(3-(6,7-dichIoro-2-(2-hydroxyacetyl)-2,3,4,5- tetrahydro- 1H-pyrido[4,3-Bbindol-9-yl)-lH-pyrazol-l-yl)ethyl)propenamide. Brown solid, yield 45%. 'H NMR (400 MHz, CD3OD) 8 7.73 - 7.67 (m, 1H), 7.21 (d, J= 16.0 Hz, 1H), 6.51 (dd, J= 24.7, 2.3 Hz, 1H), 4.65 - 4.30 (m, 6H), 3.80 - 3.44 (m, 12H), 3.13 - 2.85 (m, 4H), 2.54 - 2.34 (m, 2H).

[0781] Linker 68: 3-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)-2V-(2-(3-(6,7-dichloro-2-(2- hydroxyacetyl)-2,3,4,5-tetrahydr 0-1H-pyr ido [4 ,3-b] ind ol-9-y 1)-1H-pyrazol- 1- yl)ethyl)propenamide. Brown solid, yield 46%.1H NMR (400 MHz, CD3OD) 6 7.73 - 7.68 (m, 1H), 7.22 (d, J= 15.6 Hz, 1H), 6.51 (dd, J= 25.8, 2.3 Hz, 1H), 4.65 - 4.31 (m, 6H), 3.78 - 3.48 (m, 16H), 3.18 - 2.87 (m, 4H), 2.52 - 2.28 (m, 2H).

[0782]

[0783] Linker 69: l-amino-N-(2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-1H- pyrido[4,3-6]indol-9-yl)-1H-pyrazol-l-yl)ethyl)-3,6,9,12-tetraoxapentadecan-15-amide.

[0784] Brown solid, yield 42 %. 'H NMR (400 MHz, CD3OD) δ 7.68 (dd, J= 12.8, 2.3 Hz, 1H), 7.22 (d, J= 16.5 Hz, 1H), 6.51 (dd, J= 25.3, 2.3 Hz, 1H), 4.65 - 4.31 (m, 6H), 3.78 - 3.51 (m, 20H), 3.17 - 2.82 (m, 4H), 2.57 - 2.31 (m, 2H).

[0785] Linker 70: l-amino-N-(2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-1H- pyrido[4,3-bbindol-9-yl)-lH-pyrazol-l-yl)ethyl)-3,6,9,12,15-pentaoxaoctadecan-18-amide.

[0786] Brown solid, yield 44%. 'H NMR (400 MHz, CD3OD) 5 7.74 - 7.66 (m, 1H), 7.22 (d, J = 17.0 Hz, 1H), 6.51 (dd, J= 25.1, 2.3 Hz, 1H), 4.65 - 4.30 (m, 6H), 3.79 - 3.48 (m, 24H), 3.18 - 3.04 (m, 4H), 2.55 - 2.34 (m, 2H).

[0787] Scheme 21. The syntheses of linker 71 ethyl 2-bromoacetate

[0788] Intermediate 20

[0789] Linker 71: 7V-(2-aminoethyl)-2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-lH- pyrido[4,3- / >]indol-9-yl)-1H-pyrazol-l-yl)acetamide (XH168-108) To a solution of 3-(4, 4,5,5- tetramethyl-l,3,2-dioxaborolan-2-yl)-l / / -pyrazole (7.3 g, 37.6 mmol, 1.0 eq) in MeCN (30ml) was added K2CO3(15.6 g, 112.9 mmol, 3.0 eq) and tert-butyl 2-bromoacetate (7.5 g, 45.1 mmol, 1.2 eq). The mixture was stirred at 80 °C for 6h. After that, water was added and the mixture was extracted with ethyl acetate. The organic layer was collected, and excess solvent was removed to get intermediate 20 and used for the next step directly without further purification.

[0790] To a solution of intermediate 17 (1.05 g, 2.5 mmol, 1 eq) and intermediate 20 (2.7 g, 9.5 mmol, 3.8 eq) in dioxane (40 ml) was added KOAc (981.4 mg, 10.0 mmol, 4.0 eq) and Pd(dppf)C12CH2C12(408 mg, 0.5 mmol, 0.2 eq). The mixture was stirred at 100 °C for 6 h under N2. Ater cooling down to room temperature, the reaction mixture was concentrated under reduced pressure and purified by column chromatography (SiCh, DCM / MeOH = 10: 1) to give intermediate 21 as a brown oil, 68% yield.

[0791] To a solution of intermediate 21 (840 mg, 1.7 mmol, 1 eq) in MeOH (20 ml) and H2O (5 ml) was added LiOH (214 mg, 5.1 mmol, 3 eq). After stirred at rt for 10 min, excess MeOH was removed, resulted residue was diluted with H2O (2 mL) and the pH value was adjusted to 6 with HC1 aq. (IM) followed by purified by prep-HPLC to yield intermediate 22 as a yellow solid (456 mg, 33% in 2 steps). 'H NMR (400 MHz, CD3OD) 5 7.72 (d, J= 2.3 Hz, 1H), 7.24 (s, 1H), 6.53 (d, J= 2.3 Hz, 1H), 4.89 - 4.84 (m, 2H), 4.63 - 4.49 (m, 2H), 4.37 - 4.26 (m, 2H), 4.01 - 3.70 (m, 2H), 3.01 - 2.85 (m, 2H).

[0792] To a solution of intermediate 22 (65 mg, 0.12 mmol) in DMF (0.5 mL) were added tertbutyl (2- aminoethyl)carbamate (19.2 mg, 0.12 mmol, 1.0 equiv), l-ethyl-3-(3- dimethylaminopropyl)carbodiimide (EDCI, 34.3 mg, 0.18 mmol, 1.5 equiv), l-hydroxy-7- azabenzo-tri azole (HO At, 24.5 mg, 0.18 mmol, 1.5 equiv), and N-m ethyl morpholine (NMM, 36.4 mg, 0.36 mmol, 3.0 equiv). After being stirred for overnight atrt, the resulting mixture was purified by prep-HPLC to afford crude product. The crude product was dissolved in DCM / TFA (1 : 1, 2 mL) followed by stirred at rt for 30 min. The resulting mixture was purified by prep-HPLC to afford titled compound as yellow solid (44 mg, 63% yield). 1H NMR (400 MHz, CD3OD) 5 7.77 (s, 1H), 7.14 (s, 1H), 6.51 (s, 1H), 5.02 - 4.97 (m, 2H), 4.58 (s, 1H), 4.32 (s, 1H), 4.17 (s, 1H), 3.92 (s, 1H), 3.64 - 3.53 (m, 1H), 3.52 - 3.49 (m, 2H), ), 3.29 - 3.23 (m, 1H), 3.06 (s, 2H), 2.89 - 2.81 (m, 2H).

[0793] Linker 72 - 84 were synthesized following the same procedure for preparing linker 71.

[0794] Linker 72: N -(3-aminopropyl)-2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro- lH-pyrido[4,3- / >]indol-9-yl)-LH-pyrazol-l-yl)acetamide (xhl68-109). Yellow solid, 53% yield 'H NMR (400 MHz, CD3OD) δ 7.74 (s, 1H), 7.19 (s, 1H), 6.54 (s, 1H), 4.96 (s, 2H), 4.64 (s, 1H), 4.41 (s, 1H), 4.35 (s, 1H), 4.21 (s, 1H), 3.91 (s, 1H), 3.67 (s, 1H), 3.34 (s, 3H), 2.97 - 2.87 (m, 3H), 1.92 - 1.75 (m, 2H). Linker 75: N-(6-aminohexyl)-2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-lH- pyrido[4,3-b]indol-9-yl)-1H-pyrazol-l-yl)acetamide (xhl68-114). Yellow solid, 38% yield. 1H NMR (400 MHz, CD3OD) 8 7.73 (s, 1H), 7.20 (s, 1H), 6.54 (s, 1H), 4.93 (s, 2H), 4.71 - 4.57 (m,

[0795] 1H), 4.47 - 4.37 (m, 1H), 4.35 - 4.28 (m, 1H), 4.28 - 4.16 (m, 1H), 4.00 - 3.79 (m, 1H), 3.70 - 3.67 (m, 1H), 3.30 - 3.21 (m, 2H), 2.92 - 2.85 (m, 4H), 1.62 - 1.51 (m, 4H), 1.38 - 1.33 (m, 4H).

[0796] Linker 76: JV-(7-aminoheptyl)-2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-1H-pyrido[4,3- / ;|indol-9-yl)-l1H-pyrazol-1 -yl)acetaniide (xhl68-115). Yellow solid, 35% yield

[0797] NMR (400 MHz, CD3OD) δ 7.74 (s, 1H), 7.20 (s, 1H), 6.54 (s, 1H), 4.92 (s, 2H), 4.68 - 4.56

[0798] (m, 1H), 4.45 - 4.34 (m, 1H), 4.35 - 4.28 (m, 1H), 4.26 - 4.18 (m, 1H), 3.92 - 3.89 (m, 1H), 3.69 - 3.66 (m, 1H), 3.23 - 3.18 (m, 2H), 2.93 - 2.82 (m, 4H), 1.62 - 1.47 (m, 4H), 1.32 - 1.29 (m, 6H).

[0799] Linker 77: Af-(8-aminooctyl)-2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-1H pyrido[4,3-b]indol-9-yl)-1H-pyrazol-l-yl)acetamide (xhl 68-116). Yellow solid, 33% yield.1H NMR (400 MHz, CD3OD) 8 7.73 (s, 1H), 7.19 (s, 1H), 6.53 (s, 1H), 4.92 (s, 2H), 4.67 - 4.60 (m, 1H), 4.43 - 4.37 (m, 1H), 4.36 - 4.28 (m, 1H), 4.26 - 4.18 (m, 1H), 3.99 - 3.83 (m, 1H), 3.75 - 3.62 (m, 1H), 3.27 - 3.16 (m, 2H), 2.96 - 2.74 (m, 4H), 1.71 - 1.42 (m, 4H), 1.41 - 1.16 (m, 8H).

[0800] Linker 78: JV-(9-aminononyl)-2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-1H- pyrido[4,3-b]indol-9-yl)-1H-pyrazol-l-yl)acetamide (xhl68-117). Yellow solid, 79% yield. 1H NMR (400 MHz, CD3OD) 8 7.72 (s, 1H), 7.17 (s, 1H), 6.52 (s, 1H), 4.92 (s, 2H), 4.74 - 4.56 (m, 1H), 4.50 - 4.38 (m, 1H), 4.31 - 4.21 (m, 2H), 4.00 - 3.84 (m, 1H), 3.75 - 3.59 (m, 1H), 3.27 - 3.09 (m, 2H), 3.03 - 2.68 (m, 4H), 1.61 - 1.46 (m, 4H), 1.33 - 1.20 (m, 10H).

[0801] Linker 79: N-(10-aminodecyl)-2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-1H-pyrido|4.3-bindol-9-yl)- 1 / / -pyrazol-1 -yl)acetaniide (xhl68-118). Yellow solid, 39 mg, 47% yield. 'H NMR (400 MHz, CD3OD) 8 7.73 (s, 1H), 7.20 (s, 1H), 6.54 (s, 1H), 4.92 (s, 2H), 4.71 - 4.60 (m, 1H), 4.49 - 4.40 (m, 1H), 4.38 - 4.31 (m, 1H), 4.31 - 4.18 (m, 1H), 3.97 - 3.91 (m, 1H), 3.76 - 3.62 (m, 1H), 3.28 - 3.11 (m, 2H), 3.03 - 2.79 (m, 4H), 1.63 - 1.45 (m, 4H), 1.35 - 1.21 Linker 80: N-(2-(2-aminoethoxy)ethyl)-2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5- tetrahydro-1H-pyrido[4,3-^]indoI-9-yl)-1H-pyrazol-l-yl)acetamide (xhl68-119). Yellow solid, 53% yield. ’H NMR (400 MHz, CD3OD) 8 7.72 (s, 1H), 7.18 (s, 1H), 6.52 (s, 1H), 4.97 -

[0802] 4.96 (m, 2H), 4.72 - 4.60 (m, 1H), 4.41 - 4.35 (m, 2H), 4.29 - 4.14 (m, 1H), 3.91 - 3.88 (m, 1H),

[0803] 3.66 - 3.63 (m, 3H), 3.60 - 3.51 (m, 2H), 3.51 - 3.39 (m, 2H), 3.16 - 3.00 (m, 2H), 2.99 - 2.78

[0804] (m, 2H).

[0805] Linker 81: N -(2-(2-(2-aminoethoxy)ethoxy)ethyl)-2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-

[0806] 2.3.4.5-tetrahydro- 1 / / -pyrido|4.3-b)|indol-9-yl)-l / / -pyrazol-1 -yl)acetamide (xhl68-120).

[0807] Yellow solid, 46% yield. 'H NMR (400 MHz, CD3OD) 8 7.72 (s, 1H), 7.17 (s, 1H), 6.51 (s, 1H), 4.96 - 4.94 (m, 2H), 4.68 - 4.58 (m, 1H), 4.50 - 4.39 (m, 1H), 4.39 - 4.31 (m, 1H), 4.30 - 4.22 (m, 1H), 3.91 - 3.88 (m, 1H), 3.69 - 3.48 (m, 9H), 3.47 - 3.36 (m, 2H), 3.12 - 2.97 (m, 2H), 2.95 - 2.79 (m, 2H).

[0808] Linker 82: JV-(2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethyl)-2-(3-(6,7-dichloro-2-(2- hydroxyacetyl)-2, 3, 4, 5- tetrahydro-1H-pyrido[4,3-b]indol-9-yl)-1H-pyrazol-l-yl)acetamide (xhl68-121). Yellow solid, 30% yield. 'H NMR (400 MHz, CD3OD) 8 7.73 (s, 1H), 7.18 (s, 1H), 6.53 (s, 1H), 4.96 - 4.90 (m, 2H), 4.68 - 4.60 (m, 1H), 4.54 - 4.43 (m, 1H), 4.38 - 4.30 (m, 1H), 4.30 - 4.19 (m, 1H), 4.00 - 3.84 (m, 1H), 3.73 - 3.45 (m, 13H), 3.43 - 3.35 (m, 2H), 3.07 - 3.03 (m, 2H), 2.92 - 2.85 (m, 2H).

[0809] Linker 84: JV-(17-amino-3,6,9,12,15-pentaoxaheptadecyl)-2-(3-(6,7-dichloro-2-(2- hydroxyacetyl)-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indol-9-yl)-1H-pyrazol-l-yl)acetamide (xhl68-123) Yellow solid, 32% yield.1H NMR (400 MHz, CD3OD) 8 7.76 (s, 1H), 7.23 (s, 1H), 6.57 (s, 1H), 4.92 (s, 2H), 4.71 - 4.61 (m, 1H), 4.51 - 4.41 (m, 1H), 4.38 - 4.29 (m, 1H), 4.29 - 4.18 (m, 1H), 3.99 - 3.89 (m, 1H), 3.71 - 3.48 (m, 21H), 3.46 - 3.36 (m, 2H), 3.12 - 2.98 (m, 2H), 2.98 - 2.83 (m, 2H).

[0810] Scheme 22. The syntheses of linker 85

[0811]

[0812] Intermediate 22 Linker 85

[0813] Linker 85: 7V-(2-aminoethyl)-2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-lH- pyrido[4,3-b]indol-9-yl)-lJ / -pyrazol-l-yl)acetamide (XH168-108). To a solution of Intermediate 22 (50 mg, 0.1 mmol) in DMF (1 mL) were added tert-butyl glycinate (13.1 mg, 0.1 mmol, 1.0 equiv), l-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI, 28.7 mg, 0.15 mmol, 1.5 equiv), l-hydroxy-7-azabenzo-triazole (HOAt, 20.4 mg, 0.15 mmol, 1.5 equiv), and N- methylmorpholine (NMM, 30.3 mg, 0.3 mmol, 3.0 equiv). After being stirred at rt overnight, the resulting mixture was puritert by prep-HPLC to afford crude product. The crude product was dissolved in DCM / TFA (1 :1, 2 mL) followed by stirred at rt for 30 min. The resulting mixture was purified by prep- HPLC to afford titled compound as white solid (16 mg, 32% yield).JH NMR (400 MHz, CD3OD) 87.76 (s, 1H), 7.21 (s, 1H), 6.55 (s, 1H), 5.07 - 4.98 (m, 2H), 4.68 - 4.56 (m, 1H), 4.49 - 4.39 (m, 1H), 4.35 - 4.24 (m, 1H), 4.25 - 4.14 (m, 1H), 4.04 - 3.86 (m, 3H), 3.81 - 3.63 (m, 1H), 3.03 - 2.75 (m, 2H).

[0814] Linkers 86 - 99 were synthesized following the same procedure for preparing linker 85. Linker 86: 3-(2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-1H-pyrido[4,3- / >]indol-9-yl)-lH-pyrazol-l-yl)acetamido)propanoic acid (XH181-71). White solid, 49% yield. White solid, 49%.1H NMR (400 MHz, CD3OD) δ 7.75 (s, 1H), 7.23 (s, 1H), 6.55 (s, 1H), 4.94 (s, 2H), 4.69 - 4.59 (m, 1H), 4.49 - 4.39 (m, 1H), 4.36 - 4.26 (m, 1H), 4.26 - 4.17 (m, 1H), 4.03 - 3.87 (m, 1H), 3.83 - 3.64 (m, 1H), 3.58 - 3.40 (m, 2H), 3.02 - 2.76 (m, 2H), 2.60 - 2.45 (m, 2H).

[0815] Linker 87: 4-(2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-1H-pyrido[4,3- b ]indol-9-yl)-1H pyrazol-l-yl)acetamido)butanoic acid (XH181-74). White solid, 70% yield.

[0816] NMR (400 MHz, CD30D) 3 7.76 (s, 1H), 7.22 (s, 1H), 6.55 (s, 1H), 4.97 - 4.93 (m, 2H), 4.69 - 4.59 (m, 1H), 4.49 - 4.39 (m, 1H), 4.36 - 4.26 (m, 1H), 4.25 - 4.15 (m, 1H), 4.00 - 3.88 (m, 1H), 3.79 - 3.69 (m, 1H), 3.23 - 3.17 (m, 2H), 3.01 - 2.75 (m, 2H), 2.41 - 2.25 (m, 2H), 1.69 -

[0817] Linker 88: 5-(2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-1H-pyrido[4,3- b]indol-9-yl)-l / / -pyrazol-l-yl)acetamido)pentanoic acid (XH181-75). White solid, 47% yield. ’H NMR (400 MHz, CD3OD) δ 7.75 (s, 1H), 7.22 (s, 1H), 6.55 (s, 1H), 5.01 - 4.90 (m, 2H), 4.73 - 4.61 (m, 1H), 4.48 - 4.38 (m, 1H), 4.37 - 4.28 (m, 1H), 4.26 - 4.14 (m, 1H), 4.00 - 3.88 (m, 1H), 3.81 - 3.69 (m, 1H), 3.27 - 3.19 (m, 2H), 3.01 - 2.78 (m, 2H), 2.41 - 2.24 (m, 2H), 1.72 - 1.45 (m, 4H).

[0818]

[0819] Linker 89: 6-(2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-l1H-pyrido[4,3- b]indol-9-yl)-lH-pyrazol-l-yl)acetamido)hexanoic acid (XH181-151). Yellow solid, 25% yield. ’H NMR (400 MHz, CD3OD) δ 7.74 (s, 1H), 7.21 (s, 1H), 6.55 (s, 1H), 4.99 - 4.91 (m, 2H), 4.71 - 4.60 (m, 1H), 4.50 - 4.37 (m, 1H), 4.37 - 4.26 (m, 1H),4.27 - 4.14 (m, 1H), 4.02 - 3.85 (m, 1H), 3.79 - 3.64 (m, 1H), 3.28 - 3.15 (m, 2H), 3.03 - 2.79 (m, 2H), 2.33 - 2.14 (m, 2H), 1.69 - 1.43 (m, 4H), 1.43 - 1.24 (m, 2H).

[0820] Linker 90: 7-(2-(3-(6,7-dichloro-2-(2-hydroxyacetyI)-2,3,4,5-tetrahydro-1H-pyrido[4,3- / >]indol-9-yl)-lH-pyrazol-l-yl)acetamido)heptanoic acid (XH181-152). Yellow solid, 27% yield. Yellow solid, 27%. 'H NMR (400 MHz, CD3OD) 5 7.74 (s, 1H), 7.21 (s, 1H), 6.54 (s, 1H), 4.99 - 4.90 (m, 2H), 4.72 - 4.62 (m, 1H), 4.49 - 4.39 (m, 1H), 4.37 - 4.27 (m, 1H),4.26 - 4.12 (m, 1H), 4.03 - 3.89 (m, 1H), 3.82 - 3.65 (m, 1H), 3.27 - 3.14 (m, 2H), 3.01 - 2.81 (m, 2H), 2.33 - 2.15 (m, 2H), 1.66 - 1.42 (m, 4H), 1.41 - 1.22 (m, 4H).

[0821]

[0822] Linker 91: 8-(2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-l / / -pyrido[4,3- bdol-9-yl)- 1 / / -pyrazol- l-yl)acetamido)octanoic acid (XH181-81). White solid, 67% yield.

[0823] NMR (400 MHz, CD30D) 6 7.75 (s, 1H), 7.23 (s, 1H), 6.55 (s, 1H), 4.99 - 4.89 (m, 2H), 4.71 - 4.59 (m, 1H), 4.48 - 4.36 (m, 1H), 4.35 - 4.26 (m, 1H), 4.24 - 4.15 (m, 1H), 4.00 - 3.87 (m, 1H), 3.82 - 3.65 (m, 1H), 3.26 - 3.16 (m, 2H), 3.01 - 2.77 (m, 2H), 2.36 - 2.18 (m, 2H), 1.67 - 1.42 (m, 4H), 1.39 - 1.26 (m, 6H).

[0824] Linker 92: 9-(2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-1H-pyrido [4,3- / >]indol-9-yl)-l£Z-pyrazol-l-yl)acetamido)nonanoic acid (XH181-82). White solid, 54% yield. ’H NMR (400 MHz, CD3OD) δ 7.74 (s, 1H), 7.21 (s, 1H), 6.54 (s, 1H), 5.03 - 4.90 (m, 2H), 4.71 - 4.53 (m, 1H), 4.49 - 4.38 (m, 1H), 4.37 - 4.26 (m, 1H), 4.26 - 4.12 (m, 1H), 4.02 - 3.87 (m, 1H), 3.78 - 3.63 (m, 1H), 3.27 - 3.15 (m, 2H), 2.99 - 2.81 (m, 2H), 2.26 - 2.16 (m, 2H), 1.62 - 1.43 (m, 4H), 1.42 - 1.19 (m, 8H). Linker 93: 10-(2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-l1H-py r ido [4,3- b>]indol-9-yl)-lH-pyrazol-l-yl)acetamido)decanoic acid Yellow solid, 50%. 'HNMR (400 MHz, CD3OD) 5 7.77 (s, 1H), 7.24 (s, 1H), 6.57 (s, 1H), 5.06 - 4.93 (m, 2H), 4.74 - 4.60 (m, 1H), 4.48 - 4.38 (m, 1H), 4.39 - 4.30 (m, 1H), 4.28 - 4.14 (m, 1H), 3.99 - 3.87 (m, 1H), 3.81 - 3.69 (m, 1H), 3.28 - 3.14 (m, 2H), 3.08 - 2.78 (m, 2H), 2.38 - 2.09 (m, 2H), 1.72 - 1.40 (m, 4H), 1.39 -

[0825] Linker 95: 3-(2-(2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5-tetrahydro-1H / -pyrido[4,3- bJindol-9-yl)-1H-pyrazol-l-yl)acetamido)ethoxy)propanoic acid (XH181-153). White solid, 35% yield.1H NMR (40...

Claims

WHAT IS CLAIMS IS:

1. An OTUB 1 binder comprising a compound according to Formula (A-I) :Formula (A-I) whereinADis selected from N or CRD2;BD is selected from C=O, CH2, CH RD2, or C(RD2)2;Ring CDis absent, or selected from C3-C12cycloalkyl, 3-12-membered heterocyclic, C6- C10 aryl, and 5-10 membered heteroaryl;Ring DDis a saturated or partially unsaturated 4-12 membered heterocyclic;FD is selected from N, or CRD2;LD1is a bond, or a bivalent group selected from -O-, -NRD5-, -C(O)-, -C(O)O-, -C(O)NRD5-, -NRD5C(O)-, -OC(O)NRD5-, -NRD5C(O)O-, -NRD6C(O)NRD5-, -S(O)-, -S(O)2-, -S(O)NRD5-, - S(O)2NRD5-, C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, and C2- C6alkynylene;LD2is selected from -C(O)-, -S(O)-, -S(O)2-, -NRD5C(O)-, and C1-C6alkylene;LD3is a bond, or a bivalent group selected from C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, and C2-C6alkynylene; RD1is selected from C1-C6haloalkyl, C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2-C6heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic, where each said C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2- C6, heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic were optionally substituted with hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O)NRD7RD8, C(O)ORD7, tri(Ci-C3alkyl) silyl, C1-C6alkyl, C1-C6heteroalkyl, and 3-8 membered heterocyclic;each RD2is independently selected from hydrogen, halogen, cyano, ORD7, NRD7RD8, C1- C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6heteroalkenyl, C1-C6alkynyl, C2-C6heteroalkynyl, C3-C8cycloalkyl, and 3-8 membered heterocyclic; or two RD2groups, together with the atom(s) to which they are connected, optionally form a C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, where each said C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, together with Ring DD, can optionally form fused rings or bridged rings; each RD3is selected from hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O)ORD7, C(O)NRD7RD8, -OC(O)NRD7RD8, - NRD9C(O)NRD7RD8-, -S(O)NRD7RD8-, -S(O)2NRD7RD8-, CI-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C1-C6alkynyl, C3-C12cycloalkyl, 3-12- membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl; or two RD3groups, together with the atom(s) to which they are connected, optionally form a C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, where each said C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, together with Ring CD, optionally form fused rings or bridged rings; RD4is RD4aor RD4b; RD4;Iis selected from hydrogen, ORD7, NRD7RD8, C(O)RD7, C(O)ORD7, C(O)NRD7RD8, - OC(O)NRD7RD8, -NRD9C(O)NRD7RD8-, S(O)RD7, S(O)2RD7, S(O)NRD7RD8, S(O)2NRD7RD8, CI-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-Ci2cycloalkyl, 3-12- membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl;Ro4bis optionally a bivalent group for connection to a linker, and is selected from a bond, -O-, -N-, -C(O)-, -C(O)O-, -C(O)NRD7-, -OC(O)NRD7-, -NRD9C(O)NRD7-, -S(O)-, -S(O)2-, - S(O)NRD7-, -S(O)2NRD7-, C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, C2-C6alkynylene, C3-C12cycloalkylene, 3-12-membered heterocyclicene, C6-C10arylene, and 5-10 membered heteroarylene; each RD5and each RD6are independently selected from hydrogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, 3-12 membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl; each RD7, each RD8and each RD9are independently selected from hydrogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C12cycloalkyl, 3-12 membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl; orRD7and RD8, together with the atom(s) to which they are connected, optionally form 3-12- membered heterocyclic ring; mDis an integer of 0-8; nDis an integer of 0-8; and tautomers and pharmaceutically acceptable salts thereof.

2. An 0TUB1 binder comprising a compound according to one of Formulae (A-I-a),(A-I-b), (A-I-c) and (A-I-d):Formula (A-I-c), or Formula (A-I-d). whereinADis selected from N or CRD2;BD is selected from C=O, CH2, CH RD2, or C(RD2)2;Ring CDis absent, or selected from C3-C12cycloalkyl, 3-12-membered heterocyclic, C6- C10aryl, and 5-10 membered heteroaryl;Ring DDis a saturated or partially unsaturated 4-12 membered heterocyclic;FD is selected from N, or CRD2;LD2is selected from -C(O)-, -S(O)-, -S(O)2-, -NRD5C(O)-, and C1a-Clk6ylene;LD3is a bond, or a bivalent group selected from C1-C6alkylene, Ci-Cg haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, and C2-C6alkynylene; RD1is selected from C1-C6haloalkyl, C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2-C6heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic, where each said C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2- C6heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 memberedheterocyclic were optionally substituted with hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O)NRD7RD8, C(O)ORD7, tri(Ci-C3alkyl) silyl, C1-C6alkyl, C1-C6heteroalkyl, and 3-8 membered heterocyclic; each RD2is independently selected from hydrogen, halogen, cyano, ORD7, NRD7RD8, C1- C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6heteroalkenyl, C1-C6alkynyl, C2-C6heteroalky nyl, C3-C8cycloalkyl, and 3-8 membered heterocyclic; or two RD2groups, together with the atom(s) to which they are connected, optionally form a C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, where each said C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, together with Ring DD, can optionally form fused rings or bridged rings; each RD3is selected from hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O)ORD7, C(O)NRD7RD8, -OC(O)NRD7RD8, - NRD9C(O)NRD7RD8-, -S(O)NRD7RD8-, -S(O)2NRD7RD8-, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C1-C6alkynyl, C3-C12cycloalkyl, 3-12- membered heterocyclic, Cg-Cio aryl, and 5-10 membered heteroaryl; or two RD3groups, together with the atom(s) to which they are connected, optionally form a C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, where each said C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, together with Ring CD. optionally form fused rings or bridged rings; RD4is RD4aor RD4b;RD4ais selected from hydrogen, ORD7, NRD7RD8, C(O)RD7, C(O)ORD7, C(O)NRD7RD8, - OC(O)NRD7RD8, -NRD9C(O)NRD7RD8-, S(O)RD7, S(O)2RD7, S(O)NRD7RD8, S(O)2NRD7RD8, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, 3-12- membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl;RD4bis optionally a bivalent group for connection to a linker, and is selected firoma bond, - O-, -N-, -C(O)-, -C(O)O-, -C(O)NRD7-, -OC(O)NRD7-, -NRD9C(O)NRD7-, -S(O)-, -S(O)2-, - S(O)NRD7-, -S(O)2NRD7-, C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, C2-C6alkynylene, C3-C12cycloalkylene, 3-12-membered heterocyclicene, C6-C10arylene, and 5-10 membered heteroarylene; each RD5and each RD6are independently selected from hydrogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, 3-12 membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl;each RD7, each RD8and each RD9are independently selected from hydrogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, 3-12 membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl; or RD7and RD8, together with the atom(s) to which they are connected, optionally form 3-12- membered heterocyclic ring; mo is an integer of 0-8; no is an integer of 0-8; and tautomers and pharmaceutically acceptable salts thereof.

3. An 0TUB1 binder comprising a compound according to one of Formulae (A-I-al), (A-I-bl), (A-I-cl) and (A-I-dl):Formula (A-I-cl), or Formula (A-I-dl) whereinBD is selected from C=O, CH2, CH RD2, or C(RD2)2;Ring CDis absent, or selected from C3-C12cycloalkyl, 3-12-membered heterocyclic, Ce- Cio aryl, and 5-10 membered heteroaryl;Ring DDis a saturated or partially unsaturated 4-12 membered heterocyclic;FD is selected from N, or CRD2;LD2is selected from -C(O)-, -S(O)-, -S(O)2-, -NRD5C(O)-, and C1-C6alkylene;LD3is a bond, or a bivalent group selected from C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, and C2-C6alkynylene;RD1is selected from C1-C6haloalkyl, C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2-C6heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic, where each said C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2- C6heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic were optionally substituted with hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O)NRD7RD8, C(O)ORD7, tri(C1-C3alkyl) silyl, C1-C6alkyl, C1-C6heteroalkyl, and 3-8 membered heterocyclic; each RD2is independently selected from hydrogen, halogen, cyano, ORD7, NRD7RD8, C1- C6, alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6heteroalkenyl, C1-C6alkynyl, C2-C6heteroalkynyl, C3-C8cycloalkyl, and 3-8 membered heterocyclic; or two RD2groups, together with the atom(s) to which they are connected, optionally form a C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, where each said C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, together with Ring DD, can optionally form fused rings or bridged rings; each RD3is selected from hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O)ORD7, C(O)NRD7RD8, -OC(O)NRD7RD8, - NRD9C(O)NRD7RD8-, -S(O)NRD7RD8-, -S(O)2NRD7RD8-, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6C a2lk-Cen6yl, C1-C6alkynyl, C3-C12cycloalkyl, 3-12- membered heterocyclic, Cg-Cio aryl, and 5-10 membered heteroaryl; or two RD3groups, together with the atom(s) to which they are connected, optionally form a C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, where each said C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, together with Ring CD, optionally form fused rings or bridged rings; RD4is RD4aor RD4b;RD4ais selected from hydrogen, ORD7, NRD7RD8, C(O)RD7, C(O)ORD7, C(O)NRD7RD8, - OC(O)NRD7RD8, -NRD9C(O)NRD7RD8-, S(O)RD7, S(O)2RD7, S(O)NRD7RD8, S(O)2NRD7RD8, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6 6yl, C2-C6alkynyl, C3-C 12 cycloalkyl, 3-12- membered heterocyclic, Cg-Cio aryl, and 5-10 membered heteroaryl; RD41>is optionally a bivalent group for connection to a linker, and is selected from a bond, -O-, -N-, -C(O)-, -C(O)O-, -C(O)NRD7-, -OC(O)NRD7-, -NRD9C(O)NRD7-, -S(O)-, -S(O)2-, - S(O)NRD7-, -S(O)2NRD7-, C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, C2-C6alkynylene, C3-C12cycloalkylene, 3-12-membered heterocyclicene, C6-C10arylene, and 5-10 membered heteroarylene; each RD5and each RD6are independently selected from hydrogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, 3-12 membered heterocyclic,C6-C10aryl, and 5-10 membered heteroaryl; each RD7, each RD8and each RD9are independently selected from hydrogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, 3-12 membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl; or RD7and RD8, together with the atom(s) to which they are connected, optionally form 3-12- membered heterocyclic ring; mo is an integer of 0-8; no is an integer of 0-8; and tautomers and pharmaceutically acceptable salts thereof.

4. An 0TUB1 binder comprising a compound according to one ofFormulae (A-I-a2), (A-I-a3), (A-I-a4), (A-I-a5), (A-I-b2), (A-I-b3), (A-I-b4), (A-I-b5), (A-I-c2), (A-I-c3), (A-I-c4), (A-I-c5), (A-I-d2), (A-I-d3), (A-I-b4) and (A-I-d5):Formula (A-I-d4), Formula (A-T-d5), whereinRing CD is absent, or selected from C3-C12cycloalkyl, 3-12-membered heterocyclic, C6- C10aryl, and 5-10 membered heteroaryl;FD is selected from N, or CRD2;LD2is selected from -C(O)-, -S(O)-, -S(O)2-, -NRD3C(O)-, and C1-C6alkylene;LD3is a bond, or a bivalent group selected from C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, and C2-C6alkynylene; RD1is selected from C1-C6haloalkyl, C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2-C6heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic, where each said C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2- C6, heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic were optionally substituted with hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O)NRD7RD8, C(O)ORD7, tri(Ci-C3alkyl) silyl, C1-C6alkyl, C1-C6heteroalkyl, and 3-8 membered heterocyclic; each RD2is independently selected from hydrogen, halogen, cyano, ORD7, NRD7RD8, C1- C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6heteroalkenyl, C1-C6alkynyl, C2-C6heteroalkynyl, C3-C8cycloalkyl, and 3-8 membered heterocyclic; or two RD2groups, together with the atom(s) to which they are connected, optionally form a C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, where each said C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, together with Ring DD, can optionally form fused rings or bridged rings; each RD3is selected from hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O)ORD7, C(O)NRD7RD8, -OC(O)NRD7RD8, - NRD9C(O)NRD7RD8-, -S(O)NRD7RD8-, -S(O)2NRD7RD8-, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C1-C6alkynyl, C3-C12cycloalkyl, 3-12- membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl; or two RD3groups, together with the atom(s) to which they are connected, optionally form a C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, where each said C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, together with Ring CD, optionally form fused rings or bridged rings; RD4is RD4aor RD4b;RD4ais selected from hydrogen, ORD7, NRD7RD8, C(O)RD7, C(O)ORD7, C(O)NRD7RD8, - OC(O)NRD7RD8, -NRD9C(O)NRD7RD8-, S(O)RD7, S(O)2RD7, S(O)NRD7RD8, S(O)2NRD7RD8, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, 3-12- membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl;RD4bis optionally a bivalent group for connection to a linker, and is selected from a bond, -O-, -N-, -C(O)-, -C(O)O-, -C(O)NRD7-, -OC(O)NRD7-, -NRD9C(O)NRD7-, -S(O)-, -S(O)2-, - S(O)NRD7-, -S(O)2NRD7-, C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, C2-C6alkynylene, C3-C12cycloalkylene, 3-12-membered heterocyclicene, C6-C10arylene, and 5-10 membered heteroarylene; each RD5and each RD6are independently selected from hydrogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, 3-12 membered heterocyclic,C6-C10aryl, and 5-10 membered heteroaryl; each RD7, each RD8and each RD9are independently selected from hydrogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-Ci2cycloalkyl, 3-12 membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl; or RD7and RD8, together with the atom(s) to which they are connected, optionally form 3-12- membered heterocyclic ring; no is an integer of 0-8; and each ED1, each ED2, each ED3, each ED4, and each ED5are independently selected from null, O, CO, SO, S(O)2, NRD2, and CRD2RD2, with the proviso that no two oxygen atoms are connected to each other;OD1is selected from an integer of 1-4;OD2, OD3, OD4, and oD5are independent selected from an integer of 0-4; and tautomers and pharmaceutically acceptable salts thereof.

5. An OTUB 1 binder comprising a compound according to one of Formulae (A-I-a6),(A-I-a7), (A-I-a8), (A-I-a9), (A-I-b6), (A-I-b7), (A-I-b8), (A-I-b9), (A-I-c6), (A-I-c7), (A-I-c8), (A-I-c9), (A-I-d6), (A-I-d7), (A-I-d8), and (A-I-d9):Formula (A-I-c8), Formula (A-I-c9),Formula (A-I-d8), and Formula (A-I-d9), wherein ring CD is absent, or selected from C3-C12cycloalkyl, 3-12-membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl;FD is selected from N, or CRD2;LD2is selected from -C(O)-, -S(O)-, -S(O)2-, -NRD5C(O)-, and C1-C6alkylene;LD3is a bond, or a bivalent group selected from C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, and C2-C6alkynylene; RD1is selected from C1-C6haloalkyl, C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2-C6heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic, where each said C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2- C6, heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic were optionally substituted with hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O)NRD7RD8, C(O)ORD7, tri(Ci-C3alkyl) silyl, C1-C6alkyl, C1-C6heteroalkyl, and 3-8 membered heterocyclic; each RD2is a substituent that can attach to anywhere on the monocyclic and bicyclic rings and is independently selected from hydrogen, halogen, cyano, ORD7, NRD7RD8, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6heteroalkenyl, C1-C6alkynyl, C2-C6heteroalkynyl, C3-C8cycloalkyl, and 3-8 membered heterocyclic; or two RD2groups, together with the atom(s) to which they are connected, optionally form a C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, where each said C3-C12cycloalkyl ringor 3-12-membered heterocyclic ring, together with Ring DD, can optionally form fused rings or bridged rings; each RD3is selected from hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O)ORD7, C(O)NRD7RD8, -OC(O)NRD7RD8, - NRD9C(O)NRD7RD8-, -S(O)NRD7RD8-, -S(O)2NRD7RD8-, C1-C6alkyl,C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C1-C6alkynyl, C3-C12cycloalkyl, 3-12- membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl; or two RD3groups, together with the atom(s) to which they are connected, optionally form a C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, where each said C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, together with Ring CD. optionally form fused rings or bridged rings; RD4is RD4aor RD4b;Ro4ais selected from hydrogen, ORD7, NRD7RD8, C(O)RD7, C(O)ORD7, C(O)NRD7RD8, - OC(O)NRD7RD8, -NRD9C(O)NRD7RD8-, S(O)RD7, S(O)2RD7, S(O)NRD7RD8, S(O)2NRD7RD8, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, 3-12- membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl;RD4bis optionally a bivalent group for connection to a linker, and is selected from a bond, -O-, -N-, -C(O)-, -C(O)O-, -C(O)NRD7-, -OC(O)NRD7-, -NRD9C(O)NRD7-, -S(O)-, -S(O)2-, - S(O)NRD7-, -S(O)2NRD7-, C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, C2-C6alkynylene, C3-C12cycloalkylene, 3-12-membered heterocyclicene, C6-C10arylene, and 5-10 membered heteroarylene; each RD5and each RD6are independently selected from hydrogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, 3-12 membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl; each RD7, each RD8and each RD9are independently selected from hydrogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, 3-12 membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl; or RD7and RD8, together with the atom(s) to which they are connected, optionally form 3-12- membered heterocyclic ring;OD1is selected from an integer of 1-4;OD2, OD3, OD4, and OD5are independent selected from an integer of 0-4; mDis an integer of 0-8;nDis an integer of 0-8; and tautomers and pharmaceutically acceptable salts thereof.

6. An 0TUB1 binder according to one of claims 1 - 3, wherein Ring DDis selected from7. An OTUB1 binder according to any one of claims 1-3, wherein Ring DDis selected from:

8. An 0TUB1 binder according to any one of claim 1-3, wherein Ring DDis selected from:

9. An 0TUB1 binder according to any one of claims 1-3, wherein Ring DDis selected from:

10. An OTUB1 binder according to any one of claim 1-5, wherein ring CD is selected from azetidinyl, pyrrolidinyl, piperidinyl, and piperazinyl thiophenyl, benzothiophenyl, tetrahydrobenzothiophenyl, thiazolyl, imidazolyl, furanyl, pyrrolyl, pyrazolyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl, benzofuranyl, indolyl, and indazolyl.

11. An 0TUB1 binder according to any one of claims 1-10 wherein - LD2-RD1is selected from12. An 0TUB1 binder selected from XS154-91, XS154-130, XS154-148, XS154-114, XS154-149, XS159-13, XS159-107, XS165-30, XS159-90, XS165-53, XS165-38, XS165-33, XS165-54, XS154-184, XS165-75, XS165-77, XS165-127, XS165-106, XS165-112, XS165-97, XS165-118, XS165-110, XS165-119, XS165-123, XS165-126, XS165-120, XS165-121, XS165- 100, XS165-99, XS165-113, XS165-109, XS165-117, XS165-105, XS165-154, XS165-170, XS165-155, XS165-172, XS165-169, XS165-177, XS175-45, XS175-46, XS175-59, XS175-63, XS175-64, XS175-67, XS175-68, XS175-70, XS175-71, XS175-76, XS175-110, XS175-120, XS175-126, XS175-132, XS175-133, XS175-137, XS175-143, XS175-148, XS175-149, XS175- 158, XS175-159, XS175-160, XS175-173, XS175-174, XS175-175, XS175-176, XS175-178, XS175-179, XS175-180, XS175-186, XS186-3, XS185-4, XS186-5, XS185-6, XS185-24, XS185-29, XS185-43, XS185-46, XS185-59, XS185-64, XS185-65, XS185-66, XS185-67, XS185-69, XS185-70, XS185-71, XS185-72, XS185-75, XS185-78, XS185-86, XS185-87, XS185-90, 91, XS185-96, XS185-97, XS185-101, XS185-109, XS185-113, XS185-114, XS185- 116, XS185-118, XS185-122, XS185-131, XS185-134, XS185-135, XS185-138, XS185-140,XS185-147, XS185-149, XS185-150, XS185-171, XS190-9, XS190-27, XS190-38, XS190-44, XS190-45, XS190-46, XS190-47, XS190-48, XS190-49, XS190-50, XS190-59, XS190-68, XS190-69, XS190-70, XS190-75, XS190-75-2, XS190-77, XS190-80, XS190-81, XS190-126, XS190-127, XS190-128, XS190-129, XS190-130, XS190-137, XS190-138, XS190-144, XS190- 157, XS190-158, XS190-170, XS190-181, XS190-182, XS190-183, XS190-186, XS197-3, XS 197-4, XS 197-5, XS 197-6, XS 197- 14, XS 197-29, XS 197-38, XS 197-49, XS 197-50, XS 197- 70, XS197-71, XS197-72, XS197-73, XS197-74, XS197-85, XS197-93, XS197-94, XS197-96, XS197-133, XS197-145, XS197-176, XS197-185, XS197-186, XS209-21, XS209-22, XS209-23, XS209-24, XS209-25, XS209-26, XS209-27, XS209-28, XS209-30, XS209-38, XS209-39, XS209-40, XS209-53, XS209-54, XS209-55, XS209-57, XS209-60, XS209-62, XS209-65, XS209-74, XS209-75, XS209-92, XS209-99, XS209-100, XS209-101, XS209-122, XS209-139, XS209-140, XS209-165, XS209-166, XS209-167, XS209-168, XS209-174, XS209-175, XS209- 176, XS209-184, XS209-185, XS224-6, XS224-106, XS224-107, XS224-108, XS224-109, XS224-110, XS224-111, XS224-116, XS224-117, XS224-118, XS224-119, XS224-143, XS224- 144, XS224-145, XS224-147, XS224-148, XS224-149, XS224-150, XS224-154, XS224-155, XS224-156, XS224-157, XS224-158, XS224-159, or analogs thereof.

14. A bivalent compound comprising a ligand bound to a de-ubiquitination tag through a linker, the de-ubiquitination tag including an 0TUB1 recruiter moiety according to Formula A-IFormula (A-I) whereinADis selected from N or CRD2;BD is selected from C=O, CH2, CH RD2, or C(RD2)2;Ring CDis absent, or selected from C3-C12cycloalkyl, 3-12-membered heterocyclic, Ce- C10 aryl, and 5-10 membered heteroaryl;Ring DDis a saturated or partially unsaturated 4-12 membered heterocyclic;FD is selected from N, or CRD2;LD1is a bond, or a bivalent group selected from -O-, -NRD5-, -C(O)-, -C(O)O-, -C(O)NRD5-, -NRD5C(O)-, -OC(O)NRD5-, -NRD5C(O)O-, -NRD6C(O)NRD5-, -S(O)-, -S(O)2-, -S(O)NRD5-, - S(O)2NRD5-, C1-C8alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, and C2- C6alkynylene;LD2is selected from -C(O)-, -S(O)-, -S(O)2-, -NRD5C(O)-, and C1-C6alkylene;LD3is a bond, or a bivalent group selected from C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, and C2-C6alkynylene; RD1is selected from C1-C6haloalkyl, C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2-C6heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic, where each said C2-C6alkenyl, C2-C6alkynyl, C2-C6heteroalkenyl, C2- Ce heteroalkynyl, partially unsaturated C4-C8cycloalkyl, and partially unsaturated 4-8 membered heterocyclic were optionally substituted with hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O)NRD7RD8, C(O)ORD7, tri(Ci-C3alkyl) silyl, C1-C6alkyl, C1-C6heteroalkyl, and 3-8 membered heterocyclic; each RD2is independently selected from hydrogen, halogen, cyano, ORD7, NRD7RD8, C1- C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6heteroalkenyl, C1-C6alkynyl, C2-C6heteroalkynyl, C3-C8cycloalkyl, and 3-8 membered heterocyclic; or two RD2groups, together with the atom(s) to which they are connected, optionally form a C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, where each said C3-C12cycloalkyl ringor 3-12-membered heterocyclic ring, together with Ring DD, can optionally form fused rings or bridged rings; each RD3is selected from hydrogen, halogen, cyano, ORD7, NRD7RD8, C(O)ORD7, C(O)NRD7RD8, -OC(O)NRD7RD8, - NRD9C(O)NRD7RD8-, -S(O)NRD7RD8-, -S(O)2NRD7RD8-, C1-C6alkyl,C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C1-C6alkynyl, C3-C12cycloalkyl, 3-12- membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl; or two RD3groups, together with the atom(s) to which they are connected, optionally form a C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, where each said C3-C12cycloalkyl ring or 3-12-membered heterocyclic ring, together with Ring CD. optionally form fused rings or bridged rings; RD4is RD4aor RD4b;RD4ais selected from hydrogen, ORD7, NRD7RD8, C(O)RD7, C(O)ORD7, C(O)NRD7RD8, - OC(O)NRD7RD8, -NRD9C(O)NRD7RD8-, S(O)RD7, S(O)2RD7, S(O)NRD7RD8, S(O)2NRD7RD8, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, 3-12- membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl;RD4bis optionally a bivalent group for connection to a linker, and is selected from a bond, -O-, -N-, -C(O)-, -C(O)O-, -C(O)NRD7-, -OC(O)NRD7-, -NRD9C(O)NRD7-, -S(O)-, -S(O)2-, - S(O)NRD7-, -S(O)2NRD7-, C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, C2-C6alkynylene, C3-C12cycloalkylene, 3-12-membered heterocyclicene, C6-C10arylene, and 5-10 membered heteroarylene; each RD5and each RD6are independently selected from hydrogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, 3-12 membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl; each RD7, each RD8and each RD9are independently selected from hydrogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, 3-12 membered heterocyclic, C6-C10aryl, and 5-10 membered heteroaryl; or RD7and RD8, together with the atom(s) to which they are connected, optionally form 3-12- membered heterocyclic ring; mID is an integer of 0-8; nDis an integer of 0-8; and tautomers and pharmaceutically acceptable salts thereof.

15. The bivalent compound of claim 14, wherein the linker is a moiety according to Formula (E):Formula (E) whereinA, W and B, at each occurrence, are independently selected from null, or bivalent moietyRNR1S(O)2R , and R NR1S(O)2N(R2)R ”, whereinR and R are independently selected from a bond, optionally substituted Rr-(C1-C8alkyl), or a moiety comprising of optionally substituted C1-C8alkyl, optionally substituted C2-C8alkenyl, optionally substituted C2-C8alkynyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkoxyC1-C8alkyl, optionally substituted Ci-C8alkylaminoCi- Csalkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1-C8alkylene, optionally substituted C2-C8alkenylene, optionally substituted C2-C8alkynylene, optionally substituted C1- C8hydroxyalkylene, optionally substituted C1-C8alkoxyC1-C8alkylene, optionally substituted C1- C8alkylaminoC1-C8alkylene, optionally substituted C1-C8haloalkylene, optionally substituted 3- 10 membered carbocyclyl, optionally substituted 4-10 membered heterocyclyl, optionally substituted C3-C13fused cycloalkyl, optionally substituted C3-C13fused heterocyclyl, optionally substituted C3-C13bridged cycloalkyl, optionally substituted C3-C13bridged heterocyclyl, optionally substituted C3-C13spiro cycloalkyl, optionally substituted C3-C13spiro heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;Rris selected from optionally substituted 3-10 membered carbocyclyl, optionally substituted 4-10 membered heterocyclyl, optionally substituted C3-C13fused cycloalkyl, optionally substituted C3-C13fused heterocyclyl, optionally substituted C3-C13bridged cycloalkyl, optionally substituted C3-C13bridged heterocyclyl, optionally substituted C3-C13spiro cycloalkyl, optionally substituted C3-C13spiro heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;R1and R2are independently selected from hydrogen, optionally substituted C1-C8alkyl, optionally substituted C2-C8alkenyl, optionally substituted C2-C a8lkynyl, optionally substituted C1-C8alkoxyalkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkylaminoC1-C8alkyl, optionally substituted 3-10 membered carbocyclyl, optionally substituted 4-10 membered heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;R and R , R1and R2, R and R1, R andR2, R and R1, R andR2together with the atom to which they are connected form a 3-20 membered cycloalkyl or 4-20 membered heterocyclyl ring; m is 0 to 15; and and pharmaceutically acceptable salts and the tautomers thereof.

16. The bivalent compound according to claim 14, wherein the linker is a moiety according to Formula (E-l):Formula (E-l ), whereinR1, R2, R3and R4, at each occurrence, are independently selected from hydrogen, halogen, hydroxyl, amino, cyano, nitro, optionally substituted C1-C8alkyl, optionally substituted C2-C8alkenyl, optionally substituted C2-C8alkynyl, optionally substituted C1-C8alkoxy, optionally substituted C1-C8alkoxyalkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkylamino, and optionally substituted C1-C8alkylaminoC1-C8alkyl, optionally substituted 3-10 membered carbocyclyl, optionally substituted 3-8 membered cycloalkoxy, optionally substituted 3-10 membered carbocyclylamino, optionally substituted 4-8 membered heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl, orR1and R2, R3and R4together with the atom to which they are connected form a 3-20 membered cycloalkyl or 4-20 membered heterocyclyl ring;A, W and B, at each occurrence, are independently selected from null, or bivalent moiety selected from R -R ”, R COR ”, R CO2R ”, R C(O)N(R5)R”, R C(S)N(R5)R ”, R OR , R OC(O)R’ , R OC(O)OR , R OCONR5R”, R SR”, R SOR , R SO2R , R’SO2N(R5)R”, RN(R5)R”, RNR5COR , RNR5C(O)OR ”, R’NR5CON(R6)R , R’NR5C(S)R ”, R’NR5S(O)R , R5NR5S(O)2R”, and RNR5S(O)2N(R6)R ”, whereinR and R are independently selected from null, optionally substituted Rr-(C1-C8alkyl), or a moiety comprising of optionally substituted C1-C8alkyl, optionally substituted C2-C8alkenyl, optionally substituted C2-C8alkynyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkoxyC1-C8alkyl, optionally substituted C1-C8alkylaminoC1-C8alkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1-C6alkylene, optionally substituted C2-C8alkenylene, optionally substituted C2-C8alkynylene, optionally substituted C1- C8hydroxyalkylene, optionally substituted C1-C8alkoxyC1-C8alkylene, optionally substituted C1- C8alkylaminoC1-C8alkylene, optionally substituted C1-C8haloalkylene, optionally substituted 3- 10 membered carbocyclyl, optionally substituted 4-10 membered heterocyclyl, optionally substituted C3-C13fused cycloalkyl, optionally substituted C3-C13fused heterocyclyl, optionally substituted C3-C13bridged cycloalkyl, optionally substituted C3-C13bridged heterocyclyl, optionally substituted C3-C13spiro cycloalkyl, optionally substituted C3-C13spiro heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;Rris selected from optionally substituted 3-10 membered carbocyclyl, optionally substituted 4-10 membered heterocyclyl, optionally substituted C3-C13fused cycloalkyl, optionally substituted C3-C13fused heterocyclyl, optionally substituted C3-C13bridged cycloalkyl, optionally substituted C3-C13bridged heterocyclyl, optionally substituted C3-C13spiro cycloalkyl, optionally substituted C3-C13spiro heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;R5and R6are independently selected from hydrogen, optionally substituted C1-C8alkyl, optionally substituted C2-C8alkenyl, optionally substituted C2-Cx alkynyl, optionally substituted C1-C8alkoxyalkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkylaminoC1-C8alkyl, optionally substituted 3-10membered carbocyclyl, optionally substituted 4-10 membered heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;R and R , R5and R6, R and R5, R andR6, R and R5, R andR6together with the atom to which they are connected form a 3-20 membered cycloalkyl or 4-20 membered heterocyclyl ring; m is 0 to 15; n, at each occurrence, is 0 to 15; and o is 0 to 15.

17. The bivalent compound according to claim 14, wherein the linker is a moiety according to Formula (E-2):Formula (E-2), whereinR1and R2, at each occurrence, are independently selected from hydrogen, halogen, hydroxyl, amino, cyano, nitro, and optionally substituted C1-C8alkyl, optionally substituted Ci- C8alkoxy, optionally substituted C1-C8alkoxy C1-C6alkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkylamino, C1-C8alkylaminoC1-C8alkyl, optionally substituted 3-10 membered carbocyclyl, optionally substituted 3-8 membered cycloalkoxy, optionally substituted 3-10 membered carbocyclylamino, optionally substituted 4-10 membered heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl, orR1and R2together with the atom to which they are connected form a 3-20 membered cycloalkyl or 4-20 membered heterocyclyl ring;A and B, at each occurrence, are independently selected from null, or bivalent moiety selected from R -R ”, R COR ”, R CO2R”, R C(O)NR3R”, R C(S)NR3R”, R OR , R OC(O)R ”, R OC(O)OR , R OCON(R3)R”, R SR”, R SOR , R SO2R”, R’SO2N(R3)R”, RN(R3)R”,R’NR3COR R NR3C(O)OR’ , R NR3CON(R4)R’, R NR3C(S)R’ , R NR3S(O)R ”, R’NR3S(O)2R ”, and RNR3S(O)2N(R4)R ”, whereinR and R are independently selected from null, optionally substituted Rr-(C1-C8alkyl), or a moiety comprising of optionally substituted C1-C8alkyl, optionally substituted C2-C8alkenyl, optionally substituted C2-C8alkynyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkoxyC1-C8alkyl, optionally substituted C1-C8alkylaminoC1-C8alkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1-C8alkylene, optionally substituted C2-C8alkenylene, optionally substituted C2-C8alkynylene, optionally substituted C1- C8hydroxyalkylene, optionally substituted C1-CxalkoxyC1-Cxalkylene, optionally substituted C1- C8alkylaminoC1-C8alkylene, optionally substituted C1-C8haloalkylene, optionally substituted 3- 10 membered carbocyclyl, optionally substituted 4-10 membered heterocyclyl, optionally substituted C3-C13fused cycloalkyl, optionally substituted C3-C13fused heterocyclyl, optionally substituted C3-C13bridged cycloalkyl, optionally substituted C3-C13bridged heterocyclyl, optionally substituted C3-C13spiro cycloalkyl, optionally substituted C3-C13spiro heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;Rris selected from optionally substituted 3-10 membered carbocyclyl, optionally substituted 4-10 membered heterocyclyl, optionally substituted C3-C13fused cycloalkyl, optionally substituted C3-C13fused heterocyclyl, optionally substituted C3-C13bridged cycloalkyl, optionally substituted C3-C13bridged heterocyclyl, optionally substituted C3-C13spiro cycloalkyl, optionally substituted C3-C13spiro heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;R3and R4are independently selected from hydrogen, optionally substituted C1-C8alkyl, optionally substituted C2-C8alkenyl, optionally substituted C2-C8alkynyl, optionally substituted C1-C8alkoxyalkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkylaminoC1-C8alkyl, optionally substituted 3-10 membered carbocyclyl, optionally substituted 4-10 membered heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;R and R , R3and R4, R and R3, R and R4, R and R3, R and R4together with the atom to which they are connected form a 3-20 membered cycloalkyl or 4-20 membered heterocyclyl ring; m, at each occurrence, is 0 to 15; andn is 0 to 15.

18. The bivalent compound according to claim 14, wherein the linker is a moiety according to Formula (E-3):Formula (E-3) whereinX is selected from O, NH, and NR7;R1, R2, R3, R4, R3, and R6, at each occurrence, are independently selected from hydrogen, halogen, hydroxyl, amino, cyano, nitro, optionally substituted C1-C8alkyl, optionally substituted C2-C8 alkenyl, optionally substituted C2-C8alkynyl, optionally substituted C1-C8alkoxy, optionally substituted C1-C8alkoxy C1-C8alkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkylamino, optionally substituted C1-C8alkylaminoC1-C8alkyl, optionally substituted 3-10 membered carbocyclyl, optionally substituted 3-8 membered cycloalkoxy, optionally substituted 4-10 membered heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;A and B are independently selected from null, or bivalent moiety selected from R -R ”, R COR ”, R CO2R , RC(O)N(R8)R”, R C(S)N(R8)R”, R OR , R OC(O)R”, R OC(O)OR”, R OCON(R8)R ”, R SR ”, R SOR”, R SO2R”, R SO2N(R8)R”, RN(R8)R ”, RNR8COR”, RNR8C(O)OR”, RNR8CON(R9)R”, R’NR8C(S)R”, R’NR8S(O)R”, R’NR8S(O)2R”, and R’NR8S(O)2N(R9)R”, whereinR and R are independently selected from null, optionally substituted Rr-(C1-C6alkyl), or a moiety comprising of optionally substituted C1-C8alkyl, optionally substituted C2-C a8lkenyl, optionally substituted C2-C8alkynyl, optionally substituted C1-C8hydroxyalkyl, optionally substituted C1-C8alkoxyC1-C8alkyl, optionally substituted C1-C8alkylaminoC1-C8alkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1-C8alkylene, optionallysubstitutedC2-C8alkenylene, optionally substituted C2-C a8lkynylene, optionally substituted C1- C8hydroxy alkylene, optionally substituted C1-C8alkoxyC1-C8alkylene, optionally substituted C1- C8alkylaminoC1-C8alkylene, optionally substituted C1-C8haloalkylene, optionally substituted 3- 10 membered carbocyclyl, optionally substituted 4-10 membered heterocyclyl, optionally substituted C3-C13fused cycloalkyl, optionally substituted C3-C13fused heterocyclyl, optionally substituted C3-C13bridged cycloalkyl, optionally substituted C3-C13bridged heterocyclyl, optionally substituted C3-C13spiro cycloalkyl, optionally substituted C3-C13spiro heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;Rris selected from optionally substituted 3-10 membered carbocyclyl, optionally substituted 4-10 membered heterocyclyl, optionally substituted C3-C13fused cycloalkyl, optionally substituted C3-C13fused heterocyclyl, optionally substituted C3-C13bridged cycloalkyl, optionally substituted C3-C13bridged heterocyclyl, optionally substituted C3-C13spiro cycloalkyl, optionally substituted C3-C13spiro heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;R7, R8and R9are independently selected from hydrogen, optionally substituted C1-C8alkyl, optionally substituted C2-C8alkenyl, optionally substituted C2-C a8lkynyl, optionally substituted C1-C8alkoxyalkyl, optionally substituted C1-C8haloalkyl, optionally substituted C1- C8hydroxyalkyl, optionally substituted C1-C8alkylaminoC1-C8alkyl, optionally substituted 3-10 membered carbocyclyl, optionally substituted 4-10 membered heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;R and R , R8and R9, R and R8, R andR9, R and R8, R and R9together with the atom to which they are connected form a 3-20 membered cycloalkyl or 4-20 membered heterocyclyl ring; m, at each occurrence, is 0 to 15; n, at each occurrence, is 0 to 15; o is 0 to 15; and p is 0 to 15.

19. The bivalent compound according to claim 14, wherein the linker is a moiety according to Formula (E-2-a) or (E-3-a):Formula (E-2-a) Formula (E-3-a) whereinA is C=O or CH2, B is C=O or CH2, and m is 0-16, n is 0-6, and o is 0-6.

20. The bivalent compound according to claim 13, wherein the linker is a moiety according toFormula (E-2-b) or (E-3-b):Formula (E-2-b) Formula (E-3-b) A is -C(O)N-, B is -C(O)N-, m is 0-16, n is 0-6, o is 0-6, n-1 is 0-6, and o-l is 0-6.

21. The bivalent compound according to any one of claims claim 14 - 20 , wherein the OTUB1 recruiter moiety is selected from:

22. A bivalent compound according to any one of claims 14 -21, wherein the ligand is an AMPK ligand selected from:wherein, ♦* denotes the point of attachment to Linker.

23. A bivalent compound according to any one of claims 14 - 22, wherein the ligand is an AMPK ligand according to Formula (B-I):Formula (B-I), whereinAAis selected fromBAis selected fromdenotes the point of attachment to Linker in Formula (A);CA is selected from N or CRA4; ring DA and ring EA are independently selected from null, C3-C12cycloalkyl, 3-12- membered heterocyclyl, C6-C10aryl, and 5-10 membered heteroaryl;XAand YAare independently selected from a bond, -O-, -S-, -NRA9-, -C(O)-, -C(O)O-, - C(O)NRA9-, -O-C(O)NRA9-, -NRA10C(O)NRA9-, -S(O)-, -S(O)2-, -S(O)NRA9-, -S(O)2NRA9-, C1- C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, and C2-C6alkynylene; RA1, RA2, each RA4, each RA5, and each RA7are independently selected from H, halogen, cyano, ORA11, NRA11RA12, C(O)ORA11, C(O)NRA11RA12, S(O)2NRA11RA12, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, and 3-12 membered heterocyclyl; RA3is selected from H, halogen, cyano, ORA11, NRA11RA12, C(O)ORA11, C(O)NRA11RA12, S(O)2NRA11RA12, C1-C6alkyl, C1- hCa6loalkyl, C1h-Cet6eroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, 3-12 membered heterocyclyl, C6-C10aryl, and 5-10 membered heteroaryl; orRA1and RA2, RA2and RA3, RA3and RA4, two RA5or two RA7, together with the atoms to which they are attached, optionally form C4-C6 cycloalkyl or 5-7 membered heterocyclyl, C6-C10aryl, and 5-10 membered heteroaryl; RA6and RA8are independently selected from a bond, -O-, -NRA13-, -C(O)-, -C(O)O-, - C(O)NRA13-, -O-C(O)NRA13-, -NRA14C(O)NRA13-, -S(O)-, -S(O)2-, -S(O)NRA13-, -S(O)2NRA13-, C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, C2-C6alkynylene, C6-C10arylene, and 5-10 membered heteroarylene; each RA9, each RA10, each RA11, each RA12, each RA13, and each RA14are independently selected from H, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclyl, C6-C10aryl, and 5-10 membered heteroaryl; or RA11and RA12together with the atoms to which they are attached, optionally form 3-12 membered heterocyclyl; and mA and nA are independently selected from 0, 1, 2, 3, and 4.

24. A bivalent compound according to any one of claims 14 - 21, wherein the ligand is a cGAS ligand according to Formula (C-I):Formula (C-I) wherein denotes the point of attachment to Linker in Formula (A);** connects to either RB1Bor RB3b, with the proviso that whenconnects to RBlb,RB3is RB3a; and when ** connects to RB3b, RB1is RB1aABis selected from N and CRB4;BB is selected from N, O, S, CRB5and NRB5;CB is selected from N and C;DB is selected from N and C;XBis XBa-XBb;XBais selected from a bond, -C(O)-, -C(O)O-, -C(O)NRB6-, -S(O)-, -S(O)2-, -S(O)NRB6-, -S(O)2NRB6-;XBbis selected from a bond, C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C3-C12cycloalkylene, and 3-12 membered heterocyclylene, C6-C10arylene, and 5-10 membered heteroarylene; RB1is selected from monovalent group RB1aand bivalent group RBlb;RBlais selected from H, halogen, cyano, ORB7, NRB7RB8, C(O)ORB7, C(O)NRB7RB8, S(O)2NRB7RB8, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, 3-12 membered heterocyclyl, C6-C10aryl, and 5-10 membered heteroaryl;RBlbis connected to the Linker, and is selected from a bond, -O-, -NRB7-, -C(O)O-, - C(O)NRB7-, -S(O)2NRB7-, C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, C2-C6alkynylene, C3-C12cycloalkylene, 3-12 membered heterocyclylene, C6-C10arylene, and 5-10 membered heteroarylene; each RB2is independently selected from H, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, or two RB2, together with the atoms to which they are attached, optionally form C3-C12cycloalkyl or 3-12 membered heterocyclyl; RB3is selected from monovalent group RB3aand bivalent group RB3b;RB3ais selected from H, halogen, ORB9, NRB9RB10, C(O)ORB9, C(O)NRB9RB10, S(O)2NRB9RB10, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, and 3-12 membered heterocyclyl;RB3bis connected to the Linker, and is selected from a bond, -O-, -NRB9-, -C(O)O-, - C(O)NRB9-, -S(O)2NRB9-, C1-C6alkylene, C1-C6haloalkylene, C1-C6heteroalkylene, C2-C6alkenylene, C2-C6alkynylene, C3-C12cycloalkylene, and 3-12 membered heterocyclylene; each RB4is independently selected from H, halogen, cyano, ORB7, NRB7RB8, C(O)ORB7, C(O)NRB7RB8, S(O)2NRB7RB8, CI-C3alkyl, Ci-C3haloalkyl, Ci-C3heteroalkyl, C2-C4alkenyl, C2-C4alkynyl, C3-C6cycloalkyl, and 3-6 membered heterocyclyl, ortwo RB4, together with the atoms to which they are attached, optionally form partially unsaturated C3-C12cycloalkyl, partially unsaturated 3-12 membered heterocyclyl, C6-C ar1y0lene, or 5-10 membered heteroarylene; RB5is selected from H, halogen, C1-C3alkyl, C1-C3haloalkyl, C1-C3heteroalkyl, C3-C6cycloalkyl, and 3-6 membered heterocyclyl; RB6, each RB7, each RB8, each RB9, and each RB10, are independently selected from H, C1- C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, and 3-12 membered heterocyclyl, or RB7and RB8, and RB9and RB10together with the atoms to which they are attached, optionally form 3-12 membered heterocyclyl; mB is selected from 1 and 2; nB is selected from 0, 1, 2 and 3; andOB is selected from 0, 1, 2, 3, 4, 5 and 6.

25. A bivalent compound according to any one of claims 14 - 21, wherein the ligand is a cGAS ligand selected fromwherein, ♦* denotes the point of attachment to Linker.

26. A bivalent compound according to any one of claims 14-21, wherein the ligand is a CFTR ligand according to Formula (D-I):denotes the point of attachment to the linker,Ac and Cc are independently selected from O, S, or C(Rc8)(Rc9);Bcis C(Rc10)(Rc11) or NRc12;Arc1and Arc2are independently selected from null, C6-C1a0ryl and 5-10 membered heteroaryl;Rc1and Rc2are independently selected from H, halo, ORc13, NRc13Rc14, C(O)ORc13, C(O)NRC13RC14, S(O)2NRC13RC14, C1-C6alkyl, hCa1lo-Cal6kyl, heCte1-rCoa6lkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, and 3-12 membered heterocyclyl, orRc1and Rc2, together with the atoms to which they are attached, optionally form C3-C12cycloalkyl;Rc3and Rc6are independently selected from H, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C12cycloalkyl, and 3-12 membered heterocyclyl; each Rc4, each Rc5, and each Rc7are independently selected from H, halogen, cyano, ORc13, NRC13RC14, C(O)ORC13, C(O)NRC13RC14, S(O)2NRC13RC14, C1-C6alkyl, halCoa1l-kCy6l, Ci-C6heteroalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-Ci2cycloalkyl, and 3-12 membered heterocyclyl, two Rc4, two Rc5, or two Rc7, together with the atoms to which they are attached, optionally form partially unsaturated C3-C12cycloalkyl, partially unsaturated 3-12 membered heterocyclyl, C6-C10aryl and 5-10 membered heteroaryl;Rc8, Rc9, Rc10and Rc11are independently selected from H, halogen, ORc13, NRc13Rc14, C(O)ORc13, C(O)NRC13RC14, S(O)2NRC13RC14, C1-C6alkyl, hCa1l-oCa6lkyl, hCe1t-eCro6alkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclyl, orRc8and Rc9, and Rc10and Rc11, together with the atoms to which they are attached, optionally form C3-C12cycloalkyl and 3-12 membered heterocyclyl;Rc12is selected from H, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclyl;Rc13and Rc14are independently selected from H, C1-C6alkyl, C1-C6haloalkyl, C1-C6heteroalkyl, C3-C12cycloalkyl, and 3-12 membered heterocyclyl, orRc13and Rc14together with the atoms to which they are attached, optionally form 3-12 membered heterocyclyl; me is 0, 1, 2, or 3; nc is 0, 1, 2, or 3; and oc is 0, 1, 2, or 3.

27. A bivalent compound according to any one of claims 14-21, wherein the ligand is a CFTR ligand selected fromwherein. denotes the point of attachment to Linker.

28. A bivalent compound including an AMPK ligand selected from XF137-81 , XF137-82, XF137-83, XF137-84, XF137-85, XF137-86, XF137-87, XF137-88, XF137-89, XF137-90, XF137-91, XF137-92, XF137-93, XF137-94, QC179-047, QC179-048, QC137-049, QC179-050, QC179-051, QC179-052, QC179-053, QC179-054, QC137-055, and examples 367 - 380, or analogs thereof.

29. A bivalent compound including a cGAS ligand selected from ZD178-28, ZD178-29, ZD178-30, ZD178-31, ZD178-32, ZD178-33, ZD178-34, ZD178-35, ZD178-58-1, ZD178-58-2, ZD178-58-3, ZD178-58-4, ZD178-58-5, ZD178-58-6, ZD178-58-7, ZD178-58-8, ZD178-58-9, ZD178-58-10, ZD178-58-11, ZD178-58-12, ZD178-58-13, ZD178-58-14, ZD178-63-1, ZD178- 63-2, ZD178-63-3, ZD178-63-4, ZD178-63-5, ZD178-63-6, ZD178-63-78, ZD178-63-8, ZD178- 63-9, ZD178-63-10, ZD178-63-11, ZD178-63-12, ZD178-63-13, ZD178-63-14, ZD178-63-15, ZD178-69-1, ZD178-69-2, ZD178-69-3, ZD178-69-4, ZD178-69-5, ZD178-69-6, ZD178-69-7, ZD178-69-8, ZD178-69-9, ZD178-69-10, ZD178-69-11, ZD178-69-12, ZD178-69-13, ZD178- 69-14, ZD178-69-15, and examples 381 - 388, or analogs thereof.

30. A bivalent compound including a CFTR ligand selected from QC166-130, QC166-131,QC166-132, QC166-133, QC166-134, QC166-135, QC166-136, QC166-137, QC166-138,QC166-139, QC166-140, QC166-141, QC166-142, QC166-143, QC 166- 167, QC166-174,QC166-175, QC 166- 176, QC166-177, QC 166-178, QC 166- 179, QC166-181, QC166-182,QC166-183, QC 166- 184, QC166-185, QC179-104, QC179-105, QC 179- 106, QC 179- 107,QC179-108, QC179-109, QC179-110, QC179-111, QC179-112, QC179-113, QC179-137,QC179-138, QC179-139, QC179-140, QC179-141, QC179-154, QC179-155, QC179-156,QC179-157, QC179-158, QC192-153, QC192-154, QC192-155, QC192-156, QC192-157,QC192-184, QC192-178, QC192-179, QC192-180, QC192-181, QC192-182, or analogs thereof.31 . The bivalent compound (E)-N-(9-(2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5- tetrahydro-lH-pyrido[4,3-b]indol-9-yl)-lH-pyrazol-l-yl)acetamido)nonyl)-3-(5-(4-(4- (dimethylamino)but-2-enoyl)-2-oxopiperazin-l-yl)thiophen-2-yl)propenamide (ZD 178-29, MS7829).

32. The bivalent compound (E)-Nl-(2-(3-(6,7-dichloro-2-(2-hydroxyacetyl)-2,3,4,5- tetrahydro-lH-pyrido[4,3-b]indol-9-yl)-lH-pyrazol-l-yl)ethyl)-Nll-(2-(5-(4-(4- (dimethylamino)but-2-enoyl)-2-oxopiperazin-l-yl)thiophen-2-yl)ethyl)undecanediamide (ZD178-58-8, MS8588).

33. The bivalent compound (£)-3-(6-(l-(2,2-difluorobenzo[t / ][l,3]dioxol-5-yl)cyclopropane- l-carboxamido)-3-methylpyridin-2-yl)-A-(7-((2-(5-(4-(4-(dimethylamino)but-2-enoyl)-2- oxopiperazin- l-yl)thiophen-2-yl)ethyl)amino)-7-oxoheptyl)benzamide (QC 166- 178, MS6178).

34. A pharmaceutical composition, comprising a compound according to any one of claims 1 - 33 and a pharmaceutically acceptable carrier.

35. A pharmaceutical composition according to claim 34, wherein the compound is present in an effective amount.

36. A pharmaceutical composition according to claim 35, wherein the composition is formulated for administration by oral, parenteral intradermal, subcutaneous, topical, rectal or transdermal delivery.

37. A method of treating an AMPK-mediated disease, comprising administering to a subject in need thereof of a bivalent compound according to any one of claims 14 - 23 and 28.

38. A method of treating a cGAS-mediated disease, comprising administering to a subject in need thereof of a bivalent compound according to any one of claims 14 - 21, 24, 25 and 29.

39. A method of treating a CFTR-mediated disease, comprising administering to a subject in need thereof of a bivalent compound according to any one of claims 14 - 21, 26, 27 and 30.

40. The method of any one of claims 37 - 39, wherein the disease is selected from cystic fibrosis, breast cancer, ovarian cancer, prostate cancer, colon cancer, pancreatic cancer, bladder cancer, liver cancer and cervical cancer.

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