US2011269244A1PendingUtilityA1

Ligand-directed covalent modification of protein

47
Assignee: PETTER RUSSELL CPriority: Dec 30, 2009Filed: Dec 30, 2010Published: Nov 3, 2011
Est. expiryDec 30, 2029(~3.5 yrs left)· nominal 20-yr term from priority
C07D 405/14C12Y 207/11001G16B 15/00C07D 487/04C07D 409/12C07D 495/04C12Y 304/21098C12N 9/93C07D 491/08C07D 417/06C07D 401/04C12Q 1/485C07D 519/00C12Q 1/37C07D 403/14C12Y 603/02C07D 403/12G01N 33/68C12N 9/506C12Q 1/25C12N 9/96C07D 417/14A61P 31/14C07D 207/16A61P 43/00C07D 493/04C07D 405/12C12N 9/12C07D 401/14C07D 239/49C07D 513/04G16C 20/50G16B 15/30
47
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Claims

Abstract

The present invention relates to enzyme inhibitors. More specifically, the present invention relates to ligand-directed covalent modification of proteins; method of designing same; pharmaceutical formulation of same; and method of use.

Claims

exact text as granted — not AI-modified
1 . A method for designing a ligand that covalently binds a target protein, the method comprising:
 A) providing a structural model of a reversible ligand docked within, or in proximity to, a ligand-binding site in a target protein;   B) identifying a lysine residue of the target protein in, or in proximity to, the ligand-binding site that is less than about 15 Å from the reversible ligand when the reversible ligand is docked in, or in proximity to, the ligand-binding site;   C) producing at least a structural model of at least one ligand-warhead compound docked within, or in proximity to, the ligand-binding site wherein the ligand-warhead compound comprises the reversible ligand in step B) or a portion thereof, a warhead comprising a reactive chemical moiety, and optionally a Tether; and   D) identifying a ligand-warhead compound whose structural model allows the lysine residue in step B) to readily assume a conformation that brings the side chain primary amine group of the lysine residue within bond-forming proximity of the warhead electrophile.   
     
     
         2 . The method of  claim 1 , further comprising:
 E) forming, for the ligand-warhead compound identified in step D), a ligand-protein covalent adduct by forming a covalent bond between the side chain primary amine group of the lysine residue identified in step B) and the warhead electrophile in ligand-warhead compound identified in step D) while substantially maintaining the non-covalent interactions between the pharmacophore of the ligand and the ligand-binding site.   
     
     
         3 . The method of  claim 2 , further comprising:
 F) evaluating the conformation of the resulting ligand-protein covalent adduct formed in step E) by analyzing the global energy of the resulting conformation, or by analyzing the energy of the conformation of the Tether.   
     
     
         4 . The method of  claim 3 , wherein steps A)-F) are iterated with changes to the Tether and the global energy of the resulting conformation is less than the previous iteration. 
     
     
         5 . The method of  claim 2 , further comprising:
 F) determining whether the ligand-binding site is occluded when the covalent bond is formed between the side chain primary amine group of the lysine residue in, or in proximity to, the ligand-binding site and the warhead electrophile.   
     
     
         6 . The method of  claim 2 , wherein the covalent bond formed in step E) is formed using a computational method in which the warhead and the side chain of the lysine residue are flexible and the remainder of the structures of the ligand-warhead compound and the ligand-binding site are fixed. 
     
     
         7 . The method of  claim 1 , wherein in step B) each of the lysine residues in, or in proximity to, the ligand-binding site of the target protein, which is less than about 15 Å from the reversible ligand when the reversible ligand is docked in, or in proximity to, the ligand-binding site, is identified. 
     
     
         8 . The method of  claim 1 , wherein step C) comprises providing a plurality of models of the ligand-warhead compound, wherein the warhead is bonded to a different substitutable position of the ligand or a portion of the ligand in each model of the ligand-warhead compound, optionally with the Tether in between the warhead and the substitutable position. 
     
     
         9 . The method of  claim 1 , wherein the target protein is an identified member of an identified protein family and the lysine residue is not conserved across the identified members of the protein family. 
     
     
         10 . The method of  claim 1 , wherein the target protein is an identified member of an identified protein family and the lysine residue is conserved among more than one identified member of the identified protein family. 
     
     
         11 . The method of  claim 10 , wherein the lysine residue is conserved across the identified members of the protein family. 
     
     
         12 . The method of  claim 1 , wherein the target protein has catalytic activity. 
     
     
         13 . The method of  claim 1 , wherein the protein family is selected from the group consisting of BCL-2, Calpains, Caspases, Cathepsins, HCV, HDAC, HSP70, HSP90, IAP, Kinase, MDM2, MMP, NHR, PI3K, Phosphatase, PARP, and HIV Protease. 
     
     
         14 . The method of  claim 13 , wherein the target protein is selected from the group consisting of XIAP, cIAP1 and cIAP2, PI3Kβ/γ, PDPK1, and HCV-NS3. 
     
     
         15 . The method of  claim 1 , wherein the ligand-binding site is a ligand-binding site for a substrate or cofactor. 
     
     
         16 . The method of  claim 1 , wherein the lysine residue is not a catalytic residue. 
     
     
         17 . The method of  claim 1 , wherein the ligand-warhead compound has a structure of Formula I: 
       
         
           
           
               
               
           
         
       
       wherein
 Scaffold is
 a) a radical resulting from the removal of a hydrogen of a ligand capable of binding to, or in proximity to, the ligand-binding site; or 
 b) a portion of a pharmacophore of a ligand resulting from truncation of the pharmacophore, such that the Scaffold is capable of binding to, or in proximity to, the ligand-binding site; 
 
 Warhead is an organic moiety optionally containing one or more heteroatoms selected from O, N, and S; the organic moiety having a molecular weight of about 14 daltons to about 200 daltons; Warhead being capable of reaction with a side chain primary amine group of a lysine residue; and Warhead being attached to Scaffold through Tether; and 
 Tether is null, a bond, or a bivalent C 1 -C 15  saturated, unsaturated, straight, branched, cyclic, bicyclic, tricyclic alkyl, alkenyl, alkynyl; bridged bicyclic, heterocycle, heteroaryl, or aryl moiety; wherein optionally one or more methylene units of the hydrocarbon chain are independently replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —C(═S)—, or C(═NR 1 )—; optionally one or more hydrogens are independently replaced by heteroatoms; and optionally one or more methine groups of the C 1 -C 15  alkyl, when present, are independently replaced by 
 
       
         
           
           
               
               
           
         
         x is 0, 1, or 2; 
         y is 1, 2, or 3; and 
         R 1  is hydrogen or C 1 -C 8  alkyl; 
       
       with the proviso that the compound of Formula I is not wortmannin: 
       
         
           
           
               
               
           
         
       
       or known analogues of wortmannin that covalently modify lysine through substantially the same mechanism as wortmannin: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       or any mechanism-based irreversible inhibitors. 
     
     
         18 . The method of  claim 17 , wherein the ligand-warhead compound has a structure of Formula I′: 
       
         
           
           
               
               
           
         
       
     
     
         19 . The method of  claim 18 , wherein Warhead is a radical resulting from the removal of a hydrogen of a compound of Formula I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, I-j, I-k, I-l, I-m, I-n, I-o, I-p, I-q, I-r, I-s, and I-t: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein
 each X 1  and X 8  is independently —O—, —S—, or —NR 6 —; 
 each X 2  is independently —R 6 , —OR 6 , or —NR 6 R 7 ; 
 each X 9  is independently 
 
       
         
           
           
               
               
           
         
         each R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  is independently hydrogen or C 1 -C 6  alkyl; wherein one or more methylene groups of the C 1 -C 6  alkyl can be replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
       
       
         
           
           
               
               
           
         
         R 1  is hydrogen or C 1 -C 8  alkyl; 
         wherein optionally when proper any two of R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; and optionally X 2  and any one of R 2 , R 3 , and R 4  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; 
         A and B are each independently an optionally substituted monocyclic, bicyclic, or tricyclic aryl or heteroaryl; and 
         n is an integer from 2-4; each n 1  and n 2  is independently an integer from 0-2; n 3  is an integer from 1-2; n 4  is an integer from 1-3; and each one of n 9 , n 10 , n 11 , and n 12  is an integer from 0-1; and n 13  is an integer from 0-2, wherein when any one of the foregoing n integers is more than 1, the adjacent carbons represented by the integer can form a single or double bond. 
       
     
     
         20 . The method of  claim 19 , wherein as least one of R 2  and R 3  of the compounds of Formulas I-b and I-c is hydrogen. 
     
     
         21 . The method of  claim 19 , wherein the compound of Formula I-a, I-d, I-e, I-j, I-k, or I-l is a compound of Formula II-a, II-b, II-c, II-d, II-e, II-f, II-g, II-h, II-i, II-j, II-k, II-l, II-m, II-n, II-o, II-p, II-q, II-r, II-s, II-t, II-u, II-v, II-w, II-x, II-y, II-z, II-aa, II-bb, II-cc, II-dd, II-ee, II-ff, II-gg, II-hh, II-jj, II-kk, II-ll, II-mm, II-nn, II-oo, or II-pp; 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein
 each m is independently an integer from 0-4; 
 each m 5  is independently an integer from 0-3; 
 each m 4  is independently an integer from 0-5; 
 each n 2  is independently an integer from 0-2; 
 each R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , and R 15  is independently hydrogen or C 1 -C 6  alkyl; R z  is hydrogen, C 1 -C 6  alkyl, halogen, CF 3 , or nitro; wherein
 one or more methylene groups of the C 1 -C 6  alkyl can be replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6 alkyl, when present, can be independently replaced by 
 
 
       
         
           
           
               
               
           
         
         
           R 1  is hydrogen or C 1 -C 8  alkyl; and 
           optionally when proper any two of R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , and R 15  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group. 
         
       
     
     
         22 . The method of  claim 19 , wherein the compound of Formula I-d, or I-h is a compound of Formula III-a, III-b, III-h, or III-i; 
       
         
           
           
               
               
           
         
       
       wherein
 n 3  is an integer from 0-2; 
 each R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  is independently hydrogen or C 1 -C 6  alkyl; 
 each B 1 , B 2 , B 4 , and B 5  is independently CR 7  or N and each B 3  is NR 7 , O, or S; 
 each R z1 , R z2 , R z3 , R z4 , and R z5  is hydrogen, C 1 -C 6  alkyl, halogen, CF 3 , or nitro; 
 one or more methylene groups of the C 1 -C 6  alkyl can be optionally replaced by —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
 
       
         
           
           
               
               
           
         
         
           R 1  is hydrogen or C 1 -C 8  alkyl; and 
           optionally when proper any two of R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group. 
         
       
     
     
         23 . The method of  claim 19 , wherein the compound of Formula I-h is a compound of Formula IV-a, IV-b, IV-c, IV-d, IV-e, IV-f, IV-g, IV-h, or IV-i: 
       
         
           
           
               
               
           
         
       
       wherein any of the substitutable hydrogens on the nitrogen heterocycle of the compound can be substituted with alkyl, alkoxy, amido, acyl, acyloxy, oxoacyl, halogen. 
     
     
         24 . The method of  claim 19 , wherein the radical resulting from the removal of a hydrogen of a compound of Formula I-a, I-d, I-k, or I-m is a radical of Formula V-a, V-b, V-c, V-d, V-e, V-f, V-g, V-h, V-i, or V-j; 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein
 m 1  and m 2  are each independently an integer from 0 to 2; 
 each R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , and R 11  is independently hydrogen or C 1 -C 6  alkyl; 
 one or more methylene groups of the C 1 -C 6  alkyl can be optionally replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; 
 one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
 
       
         
           
           
               
               
           
         
         R 1  is hydrogen or C 1 -C 8  alkyl; and 
         optionally when proper any two of R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , and R 11  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group. 
       
     
     
         25 . The method of  claim 19 , wherein the compounds of Formulae I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, I-j, I-k, I-l, I-m, I-n, I-o, I-p, I-q, I-r, I-s, and I-t are selected from the group consisting of: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein any substitutable hydrogen may be substituted with the substituents as those defined by R 2 -R 8  in formulas I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, I-j, I-k, I-l, I-m, I-n, I-o, I-p, I-q, I-r, I-s, and I-t. 
     
     
         26 . The method of  claim 19 , wherein the radical resulting from the removal of a hydrogen of a compound of Formula I-a, I-d, I-k, or I-m is a radical of Formula VI-a, VI-b, VI-c, VI-d, VI-e, VI-f, VI-g, VI-h, VI-i, VI-j, VI-k, VI-l, VI-m, VI-n, VI-o, VI-p, or VI-q: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein R zz  is hydrogen, methyl, ethyl, propyl, isopropyl, cyclopropyl, —CH 2 OCH 3 , and —CH 2 CH 2 OCH 3 . 
     
     
         27 . The method of  claim 19 , wherein Scaffold is selected from the group consisting of Formulas VII, VIII, IX-a, IX-b, XI, XII, XVI, XVII, XVIII, XIX, XX, XXI, XXII, XXIII, XXXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXXVI, and XXXVII. 
     
     
         28 . The method of  claim 18 , wherein Scaffold is a radical resulting from the removal of one or more hydrogens of a compound of Formula VII: 
       
         
           
           
               
               
           
         
         wherein 
         V and W are each independently —(CR 14 R 15 ) q X 3 (CR 16 R 17 ) r —; 
         q and r are each independently 0, 1, 2, 3, or 4; 
         X 3  is —CR 18 R 19 —, or —NR 20 —; and 
         R x , R y , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 , and R 20  are each independently hydrogen or C 1 -C 6  alkyl; wherein one or more methylene groups of the C 1 -C 6  alkyl can be replaced by —NR 1 —, —O—, —C(O)—, —SO—, —C(═S)—, optionally substituted aryl or heteroaryl groups; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
       
       
         
           
           
               
               
           
         
       
       and R 1  is hydrogen or C 1 -C 8  alkyl. 
     
     
         29 . The method of  claim 28 , wherein the compound of Formula VII is a compound of Formula VII-a: 
       
         
           
           
               
               
           
         
         wherein
 V and W are each independently —(CR 14 R 15 ) q X 3 (CR 16 R 17 ) r —; 
 q and r are each independently 0, 1, 2, 3, or 4; 
 X 3  is —CR 18 R 19 —, or —NR 20 —; 
 p is 0, 1, 2, 3, or 4; 
 R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 , and R 20  are each independently hydrogen or C 1 -C 6  alkyl; wherein one or more methylene groups of the C 1 -C 6  alkyl can be replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, —C(═S)—, optionally substituted aryl or heteroaryl groups; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
 
       
       
         
           
           
               
               
           
         
         
           R 1  is hydrogen or C 1 -C 8  alkyl; and; 
           R 23  is hydrogen, C 1 -C 6  alkyl, halogen, amino, or nitro; wherein one or more methylene groups of C 1 -C 6  alkyl can be optionally replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
         
       
       
         
           
           
               
               
           
         
         
            and 
           optionally R 21  and R 23  taken together can form a 4- to 8-membered carbocyclic or heterocyclic ring. 
         
       
     
     
         30 . The method of  claim 19 , wherein the compound of Formula I′ is a compound of Formula VII-b: 
       
         
           
           
               
               
           
         
         wherein
 V and W are each independently —(CR 14 R 15 ) q X 3 (CR 16 R 17 ) r —; 
 q and r are each independently 0, 1, 2, 3, or 4; 
 X 3  is —CR 18 R 19 —, or —NR 20 —; 
 p is 0, 1, 2, 3, or 4; 
 R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 , and R 20  are each independently hydrogen or C 1 -C 6  alkyl; wherein one or more methylene groups of the C 1 -C 6  alkyl can be replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, —C(═S)—, optionally substituted aryl or heteroaryl groups; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
 
       
       
         
           
           
               
               
           
         
         
           R 1  is hydrogen or C 1 -C 8  alkyl; and; 
           R 23  is hydrogen, C 1 -C 6  alkyl, halogen, amino, or nitro; wherein one or more methylene groups of C 1 -C 6  alkyl can be optionally replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
         
       
       
         
           
           
               
               
           
         
         
            and 
           optionally R 21  and R 23  taken together can form a 4- to 8-membered carbocyclic or heterocyclic ring; 
         
         T is Tether; and 
         R wh  is Warhead. 
       
     
     
         31 . The method of  claim 30 , wherein the compound of Formula VII-b is a compound of Formula VII-h; 
       
         
           
           
               
               
           
         
       
     
     
         32 . The method of  claim 31 , wherein the compound of Formula VII-h is a compound of Formula VII-j, VII-k, VII-l, VII-m, VII-n, or VII-o: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         33 . The method of  claim 31 , wherein the compound is selected from the group consisting of: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         34 . The method of  claim 19 , wherein Scaffold is a radical resulting from the removal of a hydrogen of a compound of Formula VIII: 
       
         
           
           
               
               
           
         
       
       wherein
 X 4  is —CR 33 — or —N—; 
 s and p are each independently 0, 1, 2, 3, or 4; 
 R 12 , R 13 , R 21 , R 22 , R 24 , R 25 , R 26 , R 27 , R 28 , R 29 , R 30 , R 31 , R 32 , and R 33  are each independently hydrogen or C 1 -C 6  alkyl; 
 R 23  is hydrogen, C 1 -C 6  alkyl, halogen, amino, or nitro; wherein one or more methylene groups of C 1 -C 6  alkyl can be optionally replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
 
       
         
           
           
               
               
           
         
         R 1  is hydrogen or C 1 -C 8  alkyl; and 
         optionally R 21  and R 23  taken together can form a 4- to 8-membered carbocyclic or heterocyclic ring. 
       
     
     
         35 . The method of  claim 18 , wherein the compound of Formula I′ is a compound of Formula VIII-a or VIII-b: 
       
         
           
           
               
               
           
         
         wherein 
         X 4  is —CR 33 — or —N—; 
         s and p are each independently 0, 1, 2, 3, or 4; 
         R 12 , R 13 , R 21 , R 22 , R 24 , R 25 , R 26 , R 27 , R 28 , and R 33  are each independently hydrogen or C 1 -C 6  alkyl; 
         R 23  is hydrogen, C 1 -C 6  alkyl, halogen, amino, or nitro; wherein one or more methylene groups of C 1 -C 6  alkyl can be optionally replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
       
       
         
           
           
               
               
           
         
         R 1  is hydrogen or C 1 -C 8  alkyl; and 
         optionally R 21  and R 23  taken together can form a 4- to 8-membered carbocyclic or heterocyclic ring; and 
         Warhead is a radical resulting from the removal of a hydrogen of a compound of Formula I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, I-j, I-k, I-l, I-m, I-n, I-o, I-p, I-q, I-r, I-s, and I-t: 
       
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein
 each X 1  and X 8  is independently —O—, —S—, or —NR 6 —; 
 each X 2  is independently —R 6 , —OR 6 , or —NR 6 R 7 ; 
 each X 9  is independently 
 
       
         
           
           
               
               
           
         
         each R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  is independently hydrogen or C 1 -C 6  alkyl; wherein one or more methylene groups of the C 1 -C 6  alkyl can be replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
       
       
         
           
           
               
               
           
         
         R 1  is hydrogen or C 1 -C 8  alkyl; 
         wherein optionally when proper any two of R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; and optionally X 2  and any one of R 2 , R 3 , and R 4  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; 
         A and B are each independently an optionally substituted monocyclic, bicyclic, or tricyclic aryl or heteroaryl; and 
         n is an integer from 2-4; each n 1  and n 2  is independently an integer from 0-2; n 3  is an integer from 1-2; n 4  is an integer from 1-3; and each one of n 9 , n 10 , n 11 , and n 12  is an integer from 0-1; and n 13  is an integer from 0-2, wherein when any one of the foregoing n integers is more than 1, the adjacent carbons represented by the integer can form a single or double bond. 
       
     
     
         36 . The method of  claim 35 , wherein the compound of Formula VIII-a or VIII-b is selected from the group consisting of: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         37 . The method of  claim 19 , wherein Scaffold is a radical resulting from the removal of a hydrogen of a compound of Formula IX-a or IX-b, 
       
         
           
           
               
               
           
         
       
       wherein
 X 5  is —O—, —CR 42 R 43 — or —NR 42 —; 
 R 12 , R 13 , R 27 , R 28 , R 29 , R 30 , R 31 , R 32 , R 33 , R 34 , R 35 , R 36 , R 37 , R 38 , R 39 , R 40 , R 41 , R 42 , and R 43  are each independently hydrogen or C 1 -C 6  alkyl; wherein one or more methylene groups of C 1 -C 6  alkyl can be optionally replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
 
       
         
           
           
               
               
           
         
         R 1  is hydrogen or C 1 -C 8  alkyl; 
         D, E, F, G, and H are each independently optionally substituted aryl or heteroaryl; wherein F and G are fused together to form a bicyclic optionally substituted aryl or heteroaryl. 
       
     
     
         38 . The method of  claim 18 , wherein the compound of Formula I′ is a compound of Formula IX-c or IX-d: 
       
         
           
           
               
               
           
         
       
       wherein
 R 12 , R 13 , R 31 , are each independently hydrogen or C 1 -C 6  alkyl; wherein one or more methylene groups of C 1 -C 6  alkyl can be optionally replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
 
       
         
           
           
               
               
           
         
         R 1  is hydrogen or C 1 -C 8  alkyl; 
         F, G, and H are each independently optionally substituted aryl or heteroaryl; wherein F and G are fused together to form a bicyclic optionally substituted aryl or heteroaryl; 
         T is Tether; 
         R wh  is Warhead which is a radical resulting from the removal of a hydrogen of a compound of Formula I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, I-j, I-k, I-l, I-m, I-n, I-o, I-p, I-q, I-r, I-s, and I-t: 
       
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein
 each X 1  and X 8  is independently —O—, —S—, or —NR 6 —; 
 each X 2  is independently —R 6 , —OR 6 , or —NR 6 R 7 ; 
 each X 9  is independently 
 
       
         
           
           
               
               
           
         
         each R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  is independently hydrogen or C 1 -C 6  alkyl; wherein one or more methylene groups of the C 1 -C 6  alkyl can be replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
       
       
         
           
           
               
               
           
         
         R 1  is hydrogen or C 1 -C 8  alkyl; 
         wherein optionally when proper any two of R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; and optionally X 2  and any one of R 2 , R 3 , and R 4  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; 
         A and B are each independently an optionally substituted monocyclic, bicyclic, or tricyclic aryl or heteroaryl; and 
         n is an integer from 2-4; each n 1  and n 2  is independently an integer from 0-2; n 3  is an integer from 1-2; n 4  is an integer from 1-3; and each one of n 9 , n 10 , n 11 , and n 12  is an integer from 0-1; and n 13  is an integer from 0-2, wherein when any one of the foregoing n integers is more than 1, the adjacent carbons represented by the integer can form a single or double bond. 
       
     
     
         39 . The method of  claim 18 , wherein the compound of Formula I′ is a compound of Formula XI: 
       
         
           
           
               
               
           
         
       
       wherein
 p is an integer from 0 to 4, u is an integer from 1 to 4; 
 B 6  and B 7  are each independently CR 7  or N; 
 R 69  is hydrogen, C 1 -C 6  alkyl, halogen, amino, nitro, or —NH(CO)NR 78 R 79 ; 
 R 70  is hydrogen, C 1 -C 6  alkyl, halogen, amino, nitro; 
 R 7 , R 71 , R 72 , R 73 , R 74 , R 75 , R 76 , R 77 , R 78 , and R 79  are each independently hydrogen or C 1 -C 6  alkyl; R 1  is hydrogen or C 1 -C 8  alkyl; wherein one or more methylene groups of the C 1 -C 6  alkyl can be replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
 
       
         
           
           
               
               
           
         
         optionally R 78 , and R 79  taken together form a 4- to 8-membered carbocyclic or heterocyclic ring; 
         T is Tether; and 
         R wh  is Warhead which is a radical resulting from the removal of a hydrogen of a compound of Formula I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, I-j, I-k, I-l, I-m, I-n, I-o, I-p, I-q, I-r, I-s, and I-t: 
       
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein
 each X 1  and X 8  is independently —O—, —S—, or —NR 6 —; 
 each X 2  is independently —R 6 , —OR 6 , or —NR 6 R 7 ; 
 each X 9  is independently 
 
       
         
           
           
               
               
           
         
         each R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  is independently hydrogen or C 1 -C 6  alkyl; wherein one or more methylene groups of the C 1 -C 6  alkyl can be replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
       
       
         
           
           
               
               
           
         
         R 1  is hydrogen or C 1 -C 8  alkyl; 
         wherein optionally when proper any two of R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; and optionally X 2  and any one of R 2 , R 3 , and R 4  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; 
         A and B are each independently an optionally substituted monocyclic, bicyclic, or tricyclic aryl or heteroaryl; and 
         n is an integer from 2-4; each n 1  and n 2  is independently an integer from 0-2; n 3  is an integer from 1-2; n 4  is an integer from 1-3; and each one of n 9 , n 10 , n 11 , and n 12  is an integer from 0-1; and n 13  is an integer from 0-2, wherein when any one of the foregoing n integers is more than 1, the adjacent carbons represented by the integer can form a single or double bond. 
       
     
     
         40 . The method of  claim 39 , wherein the compound of Formula XI is a compound of Formula XI-a, XI-b, or XI-c: 
       
         
           
           
               
               
           
         
       
     
     
         41 . The method of  claim 40 , wherein the compound of Formula XI-a, XI-b or XI-c is a compound of Formula XI-d, XI-e, XI-f, XI-g, XI-h, XI-i, or XI-j: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         42 . The method of  claim 41 , wherein the compound of Formula XI-d, XI-e, XI-f, XI-g, XI-h, XI-i, or XI-j is a compound of Formula XI-k, XI-l, XI-m, XI-n, XI-o, XI-p, or XI-q; 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein
 R 88 , and R 89  are each independently hydrogen or C 1 -C 6  alkyl; R 1  is hydrogen or C 1 -C 8  alkyl; 
 one or more methylene groups of the C 1 -C 6  alkyl can be replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; 
 one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
 
       
         
           
           
               
               
           
         
       
       and
 n 5  is an integer from 0 to 3. 
 
     
     
         43 . The method of  claim 42 , wherein the compound of Formula XI-d, XI-e, XI-f, XI-g, XI-h, XI-i, or XI-j is a compound of Formula XI-r, XI-s, XI-t, XI-u, XI-v, XI-w, or XI-x; 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         44 . The method of  claim 41 , wherein the compound of Formula XI-d, XI-e, XI-f, XI-g, XI-h, XI-i, or XI-j is a compound of Formula XI-y, XI-z, XI-aa, or XI-bb; 
       
         
           
           
               
               
           
         
       
     
     
         45 . The method of  claim 41 , wherein the compound of Formula XI-d, XI-e, XI-f, XI-g, XI-h, XI-i, or XI-j is a compound of Formula XI-cc, XI-dd, XI-ee, or XI-ff: 
       
         
           
           
               
               
           
         
       
     
     
         46 . The method of  claim 39 , wherein the compound of Formula XI is selected from the group consisting of: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         47 . The method of  claim 18 , wherein the compound of Formula I′ is a compound of Formula XII: 
       
         
           
           
               
               
           
         
       
       wherein
 R 1  and R 2  are each independently hydrogen or C 1 -C 8  alkyl; wherein one or more methylene groups of the C 1 -C 6  alkyl can be replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
 
       
         
           
           
               
               
           
         
         T is Tether; 
         R wh  is Warhead which is a radical resulting from the removal of a hydrogen of a compound of Formula I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, I-j, I-k, I-l, I-m, I-n, I-o, I-p, I-q, I-r, I-s, and I-t: 
       
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein
 each X 1  and X 8  is independently —O—, —S—, or —NR 6 —; 
 each X 2  is independently —R 6 , —OR 6 , or —NR 6 R 7 ; 
 each X 9  is independently 
 
       
         
           
           
               
               
           
         
         each R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  is independently hydrogen or C 1 -C 6  alkyl; wherein one or more methylene groups of the C 1 -C 6  alkyl can be replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
       
       
         
           
           
               
               
           
         
         R 1  is hydrogen or C 1 -C 8  alkyl; 
         wherein optionally when proper any two of R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; and optionally X 2  and any one of R 2 , R 3 , and R 4  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; 
         A and B are each independently an optionally substituted monocyclic, bicyclic, or tricyclic aryl or heteroaryl; and 
         n is an integer from 2-4; each n 1  and n 2  is independently an integer from 0-2; n 3  is an integer from 1-2; n 4  is an integer from 1-3; and each one of n 9 , n 10 , n 11 , and n 12  is an integer from 0-1; and n 13  is an integer from 0-2, wherein when any one of the foregoing n integers is more than 1, the adjacent carbons represented by the integer can form a single or double bond. 
       
     
     
         48 . The method of  claim 18 , wherein the compound of Formula I′ is a compound of Formula XXXVI: 
       
         
           
           
               
               
           
         
         wherein 
         R v  is H, optionally substituted C 1 -C 3  branched or straight chain alkyl, or optionally substituted C 1 -C 3  branched or straight chain acyl; 
         T is Tether; and 
         R wh  is Warhead which is a radical resulting from the removal of a hydrogen of a compound of Formula I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, I-j, I-k, I-l, I-m, I-n, I-o, I-p, I-q, I-r, I-s, and I-t: 
       
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein
 each X 1  and X 8  is independently —O—, —S—, or —NR 6 —; 
 each X 2  is independently —R 6 , —OR 6 , or —NR 6 R 7 ; 
 each X 9  is independently 
 
       
         
           
           
               
               
           
         
         each R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  is independently hydrogen or C 1 -C 6  alkyl; wherein one or more methylene groups of the C 1 -C 6  alkyl can be replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
       
       
         
           
           
               
               
           
         
         R 1  is hydrogen or C 1 -C 8  alkyl; 
         wherein optionally when proper any two of R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; and optionally X 2  and any one of R 2 , R 3 , and R 4  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; 
         A and B are each independently an optionally substituted monocyclic, bicyclic, or tricyclic aryl or heteroaryl; and 
         n is an integer from 2-4; each n 1  and n 2  is independently an integer from 0-2; n 3  is an integer from 1-2; n 4  is an integer from 1-3; and each one of n 9 , n 10 , n 11 , and n 12  is an integer from 0-1; and n 13  is an integer from 0-2, wherein when any one of the foregoing n integers is more than 1, the adjacent carbons represented by the integer can form a single or double bond. 
       
     
     
         49 . The method of  claim 18 , wherein the compound of Formula I′ is selected from the group consisting of: 
       
         
           
           
               
               
           
         
       
     
     
         50 . The method of  claim 19 , wherein Scaffold is a radical resulting from the removal of a hydrogen of a compound of Formula XVI-a, XVI-b, or XVI-c: 
       
         
           
           
               
               
           
         
       
       wherein
 R 90 , R 91 , R 92 , R 93 , R 94 , R 95 , R 96 , R 97 , R 98 , R 99 , R 100 , R 102 , R 104 , R 105 , R 106 , R 107 , R 108 , R 109 , R 110 , R 111 , R 112 , R 113 , and R 114  are each independently hydrogen or C 1 -C 6  alkyl; wherein one or more methylene groups of C 1 -C 6  alkyl can be optionally replaced by —NR 1 —, —O—, —C(O)—, —SO—, —SO 2 —, or —C(═S)—; 
 R 103  is hydrogen, C 1 -C 6  alkyl, or C 2 -C 8  alkenyl; 
 one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
 
       
         
           
           
               
               
           
         
         R 1  is hydrogen or C 1 -C 8  alkyl; 
         each R 101  and R 101  is independently hydrogen, C 1 -C 6  alkyl, C 2 -C 8  alkenyl, halogen, amino, nitro, optionally substituted aryl or heteroaryl; and 
         n 6  and n 7  are each independently integer from 0 to 4; and 
         n 8  is an integer from 0 to 2. 
       
     
     
         51 . The method of  claim 18 , wherein the compound of Formula I′ is a compound of Formula XVI-d, XVI-e, or XVI-f: 
       
         
           
           
               
               
           
         
       
       wherein
 R 90  and R 114  are each independently hydrogen or C 1 -C 6  alkyl; 
 R 103  is hydrogen, C 1 -C 6  alkyl, or C 2 -C 8  alkenyl; 
 R 101  is hydrogen, C 1 -C 6  alkyl, C 2 -C 8  alkenyl, halogen, amino, nitro, optionally substituted aryl or heteroaryl; and 
 n 6  is an integer from 0 to 4; 
 n 8  is an integer from 0 to 2; 
 wherein one or more methylene groups of C 1 -C 6  alkyl can be optionally replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; and 
 R wh  is Warhead which is a radical resulting from the removal of a hydrogen of a compound of Formula I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, I-j, I-k, I-l, I-m, I-n, I-o, I-p, I-q, I-r, I-s, and I-t: 
 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein
 each X 1  and X 8  is independently —O—, —S—, or —NR 6 —; 
 each X 2  is independently —R 6 , —OR 6 , or —NR 6 R 7 ; 
 each X 9  is independently 
 
       
         
           
           
               
               
           
         
         each R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  is independently hydrogen or C 1 -C 6  alkyl; wherein one or more methylene groups of the C 1 -C 6  alkyl can be replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
       
       
         
           
           
               
               
           
         
         R 1  is hydrogen or C 1 -C 8  alkyl; 
         wherein optionally when proper any two of R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; and optionally X 2  and any one of R 2 , R 3 , and R 4  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; 
         A and B are each independently an optionally substituted monocyclic, bicyclic, or tricyclic aryl or heteroaryl; and 
       
       n is an integer from 2-4; each n 1  and n 2  is independently an integer from 0-2; n 3  is an integer from 1-2; n 4  is an integer from 1-3; and each one of n 9 , n 10 , n 11 , and n 12  is an integer from 0-1; and n 13  is an integer from 0-2, wherein when any one of the foregoing n integers is more than 1, the adjacent carbons represented by the integer can form a single or double bond; and
 T is Tether. 
 
     
     
         52 . The method of  claim 51 , wherein the compound of Formula XVI-d, XVI-e, or XVI-f is a compound of Formula XVI-g, XVI-h, or XVI-i: 
       
         
           
           
               
               
           
         
       
     
     
         53 . The method of  claim 18 , wherein the compound of Formula XVI-d, XVI-e, or XVI-f is selected from the group consisting of: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         54 . The method of  claim 18  wherein the compound of Formula I′ is a compound of Formula XXII-a, Formula XXII-b, or Formula XXII-c: 
       
         
           
           
               
               
           
         
         wherein 
         n, m, p, and q for Formula XXII-a and Formula XXII-b are each independently 0, 1, 2, 3; provided that n and q are not 0 at the same time, and m and q are not 0 at the same time; 
         T is tether; 
         R wh  is Warhead and is a radical resulting from the removal of a hydrogen of a compound of Formula I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, I-j, I-k, I-l, I-m, I-n, I-o, I-p, I-q, I-r, I-s, and I-t: 
       
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein
 each X 1  and X 8  is independently —O—, —S—, or —NR 6 —; 
 each X 2  is independently —R 6 , —OR 6 , or —NR 6 R 7 ; 
 each X 9  is independently 
 
       
         
           
           
               
               
           
         
         each R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  is independently hydrogen or C 1 -C 6  alkyl; wherein one or more methylene groups of the C 1 -C 6  alkyl can be replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
       
       
         
           
           
               
               
           
         
         R 1  is hydrogen or C 1 -C 8  alkyl; 
         wherein optionally when proper any two of R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; and optionally X 2  and any one of R 2 , R 3 , and R 4  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; 
         A and B are each independently an optionally substituted monocyclic, bicyclic, or tricyclic aryl or heteroaryl; and 
         n is an integer from 2-4; each n 1  and n 2  is independently an integer from 0-2; n 3  is an integer from 1-2; n 4  is an integer from 1-3; and each one of n 9 , n 10 , n 11 , and n 12  is an integer from 0-1; and n 13  is an integer from 0-2, wherein when any one of the foregoing n integers is more than 1, the adjacent carbons represented by the integer can form a single or double bond; 
         A 2  is an optionally substituted ring selected from a 4-8 membered saturated or partially unsaturated heterocyclic ring having one or two heteroatoms independently selected from nitrogen, oxygen, or sulfur, or a 5-10 membered saturated or partially unsaturated bridged bicyclic heterocyclic ring having at least one nitrogen, at least one oxygen, and optionally 1-2 additional heteroatoms independently selected from nitrogen, oxygen, or sulfur; 
         B′ is an optionally substituted group selected from phenyl, an 8- to 10-membered bicyclic aryl ring, a 5- to 6-membered heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an 8- to 10-membered bicyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; or -T-Rwh; and 
         C 2  is hydrogen or an optionally substituted ring selected from a 3- to 7-membered saturated or partially unsaturated carbocyclic ring, a 7- to 10-membered saturated or partially unsaturated bicyclic carbocyclic ring, a 4- to 7-membered saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, a 7- to 10-membered saturated or partially unsaturated bicyclic heterocyclic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, phenyl, an 8- to 10-membered bicyclic aryl ring, a 5- to 6-membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an 8- to 10-membered bicyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. 
       
     
     
         55 . The method of  claim 54 , wherein the compound of Formula XXII-a, XXII-b, XXII-c is selected from the group consisting of: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         56 . The method of  claim 18 , wherein the compound of Formula I′ is a compound of Formula XXIII: 
       
         
           
           
               
               
           
         
         wherein: 
         R wh  is Warhead which is a radical resulting from the removal of a hydrogen of a compound of Formula I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, I-j, I-k, I-l, I-m, I-n, I-o, I-p, I-q, I-r, I-s, and I-t: 
       
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein
 each X 1  and X 8  is independently —O—, —S—, or —NR 6 —; 
 each X 2  is independently —R 6 , —OR 6 , or —NR 6 R 7 ; 
 each X 9  is independently 
 
       
         
           
           
               
               
           
         
         each R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  is independently hydrogen or C 1 -C 6  alkyl; wherein one or more methylene groups of the C 1 -C 6  alkyl can be replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
       
       
         
           
           
               
               
           
         
         R 1  is hydrogen or C 1 -C 8  alkyl; 
         wherein optionally when proper any two of R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; and optionally X 2  and any one of R 2 , R 3 , and R 4  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; 
         A and B are each independently an optionally substituted monocyclic, bicyclic, or tricyclic aryl or heteroaryl; and 
         n is an integer from 2-4; each n 1  and n 2  is independently an integer from 0-2; n 3  is an integer from 1-2; n 4  is an integer from 1-3; and each one of n 9 , n 10 , n 11 , and n 12  is an integer from 0-1; and n 13  is an integer from 0-2, wherein when any one of the foregoing n integers is more than 1, the adjacent carbons represented by the integer can form a single or double bond; 
         R 201  is hydrogen or C 1-6  alkyl; 
         R 202  is hydrogen or an optionally substituted group selected from C 1-6  alkyl, C 1-6  alkoxy, or (C 1-6  alkylene)-R 203 ; or 
         R 201  and R 202  are taken together with the intervening carbon to form an optionally substituted ring selected from a 3- to 7-membered carbocyclic ring or a 4- to 7-membered heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur; 
         R 203  is a 3- to 7-membered saturated or partially unsaturated carbocyclic ring, a 7- to 10-membered saturated or partially unsaturated bicyclic carbocyclic ring, a 4- to 7-membered saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, a 7- to 10-membered saturated or partially unsaturated bicyclic heterocyclic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, phenyl, a 8- to 10-membered bicyclic aryl ring, a 5- to 6-membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or a 8- to 10-membered bicyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; and 
         Ring A 6  is absent or an optionally substituted group selected from a 4- to 7-membered heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or a 5- to 6-membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur. 
       
     
     
         57 . The method of  claim 56 , where the compound of Formula XXIII is selected from the group consisting of: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         58 . The method of  claim 18 , wherein the compound of Formula I′ is a compound of Formula XXIV-a or Formula XXIV-b: 
       
         
           
           
               
               
           
         
         wherein 
         R wh  is Warhead which is a radical resulting from the removal of a hydrogen of a compound of Formula I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, I-j, I-k, I-l, I-m, I-n, I-o, I-p, I-q, I-r, I-s, and I-t: 
       
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein
 each X 1  and X 8  is independently —O—, —S—, or —NR 6 —; 
 each X 2  is independently —R 6 , —OR 6 , or —NR 6 R 7 ; 
 each X 9  is independently 
 
       
         
           
           
               
               
           
         
         each R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  is independently hydrogen or C 1 -C 6  alkyl; wherein one or more methylene groups of the C 1 -C 6  alkyl can be replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
       
       
         
           
           
               
               
           
         
         R 1  is hydrogen or C 1 -C 8  alkyl; 
         wherein optionally when proper any two of R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; and optionally X 2  and any one of R 2 , R 3 , and R 4  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; 
         A and B are each independently an optionally substituted monocyclic, bicyclic, or tricyclic aryl or heteroaryl; and 
         n is an integer from 2-4; each n 1  and n 2  is independently an integer from 0-2; n 3  is an integer from 1-2; n 4  is an integer from 1-3; and each one of n 9 , n 10 , n 11 , and n 12  is an integer from 0-1; and n 13  is an integer from 0-2, wherein when any one of the foregoing n integers is more than 1, the adjacent carbons represented by the integer can form a single or double bond; 
         R 204  is an hydrogen or an optionally substituted group selected from C 1-6  aliphatic, —(CH 2 ) m -(3- to 7-membered saturated or partially unsaturated carbocyclic ring), —(CH 2 ) m -(7- to 10-membered saturated or partially unsaturated bicyclic carbocyclic ring), —(CH 2 ) m -(4- to 7-membered saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur), —(CH 2 ) m -(7- to 10-membered saturated or partially unsaturated bicyclic heterocyclic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur), —(CH 2 ) m -phenyl, —(CH 2 ) m -(8- to 10-membered bicyclic aryl ring), —(CH 2 ) m -(5- to 6-membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur), or —(CH 2 ) m -(8- to 10-membered bicyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur); 
         each R 205  and R 206  is independently —R″, halogen, —NO 2 , —CN, —OR″, —SR″, —N(R″) 2 , —C(O)R″, —CO 2 R″, —C(O)C(O)R″, —C(O)CH 2 C(O)R″, —S(O)R″, —S(O) 2 R″, —C(O)N(R″) 2 , —SO 2 N(R″) 2 , —OC(O)R″, —N(R″)C(O)R″, —N(R″)N(R″) 2 , —N(R″)C(═NR″)N(R″) 2 , —C(═NR″)N(R″) 2 , —C═NOR″, —N(R″)C(O)N(R″) 2 , —N(R″)SO 2 N(R″) 2 , —N(R″)SO 2 R″, or —OC(O)N(R″) 2 ; 
         each R″ is independently hydrogen or an optionally substituted group selected from C 1-6  aliphatic, a 3- to 7-membered saturated or partially unsaturated carbocyclic ring, a 7- to 10-membered saturated or partially unsaturated bicyclic carbocyclic ring, a 4- to 7-membered saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, a 7- to 10-membered saturated or partially unsaturated bicyclic heterocyclic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, phenyl, an 8- to 10-membered bicyclic aryl ring, a 5- to 6-membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an 8- to 10-membered bicyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; or 
         two R″ groups on the same nitrogen are taken together with the nitrogen to which they are attached to form an optionally substituted 5-8 membered saturated, partially unsaturated, or aromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; 
       
       m is an integer from 0 to 6, inclusive;
 each n for Formula XXIV-a or Formula XXIV-b is independently 0, 1, or 2; and 
 Ring A 5  is an optionally substituted 6-membered heterocyclic or heteroaryl ring having 1-2 nitrogens. 
 
     
     
         59 . The method of  claim 58 , wherein the compound of Formula XXIV-a or Formula XXIV-b is selected from the group consisting of 
       
         
           
           
               
               
           
         
       
     
     
         60 . The method of  claim 18 , wherein the compound of Formula I′ is a compound of Formula XXV: 
       
         
           
           
               
               
           
         
         wherein 
         R wh  is Warhead which is a radical resulting from the removal of a hydrogen of a compound of Formula I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, I-j, I-k, I-l, I-m, I-n, I-o, I-p, I-q, I-r, I-s, and I-t: 
       
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein
 each X 1  and X 8  is independently —O—, —S—, or —NR 6 —; 
 each X 2  is independently —R 6 , —OR 6 , or —NR 6 R 7 ; 
 each X 9  is independently 
 
       
         
           
           
               
               
           
         
         each R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  is independently hydrogen or C 1 -C 6  alkyl; wherein one or more methylene groups of the C 1 -C 6  alkyl can be replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
       
       
         
           
           
               
               
           
         
         R 1  is hydrogen or C 1 -C 8  alkyl; 
         wherein optionally when proper any two of R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; and optionally X 2  and any one of R 2 , R 3 , and R 4  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; 
         A and B are each independently an optionally substituted monocyclic, bicyclic, or tricyclic aryl or heteroaryl; and 
         n is an integer from 2-4; each n 1  and n 2  is independently an integer from 0-2; n 3  is an integer from 1-2; n 4  is an integer from 1-3; and each one of n 9 , n 10 , n 11 , and n 12  is an integer from 0-1; and n 13  is an integer from 0-2, wherein when any one of the foregoing n integers is more than 1, the adjacent carbons represented by the integer can form a single or double bond; 
         each R 205  and R 206  is independently —R″, halogen, —NO 2 , —CN, —OR″, —SR″, —N(R″) 2 , —C(O)R″, —CO 2 R″, —C(O)C(O)R″, —C(O)CH 2 C(O)R″, —S(O)R″, —S(O) 2 R″, —C(O)N(R″) 2 , —SO 2 N(R″) 2 , —OC(O)R″, —N(R″)C(O)R″, —N(R″)N(R″) 2 , —N(R″)C(═NR″)N(R″) 2 , —C(═NR″)N(R″) 2 , —C═NOR″, —N(R″)C(O)N(R″) 2 , —N(R″)SO 2 N(R″) 2 , —N(R″)SO 2 R″, or —OC(O)N(R″) 2 ; 
         each R″ is independently hydrogen or an optionally substituted group selected from C 1-6  aliphatic, a 3- to 7-membered saturated or partially unsaturated carbocyclic ring, a 7- to 10-membered saturated or partially unsaturated bicyclic carbocyclic ring, a 4- to 7-membered saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, a 7- to 10-membered saturated or partially unsaturated bicyclic heterocyclic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, phenyl, an 8- to 10-membered bicyclic aryl ring, a 5- to 6-membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an 8- to 10-membered bicyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; or 
         optionally, two R″ groups on the same nitrogen are taken together with the nitrogen to which they are attached to form an optionally substituted 5-8 membered saturated, partially unsaturated, or aromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; 
         m is an integer from 0 to 6, inclusive; 
         each n for Formula XXV is independently 0, 1, or 2; and 
         Ring A 5  is an optionally substituted 6-membered heterocyclic or heteroaryl ring having 1-2 nitrogens. 
       
     
     
         61 . The method of  claim 60 , wherein the compound of Formula XXV is selected from the group consisting of: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         62 . The method of  claim 18 , wherein the compound of Formula I′ is a compound of Formula XXVII: 
       
         
           
           
               
               
           
         
         wherein: 
         T is Tether; 
         R wh  is Warhead which is a radical resulting from the removal of a hydrogen of a compound of Formula I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, I-j, I-k, I-l, I-m, I-n, I-o, I-p, I-q, I-r, I-s, and I-t: 
       
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein
 each X 1  and X 8  is independently —O—, —S—, or —NR 6 —; 
 each X 2  is independently —R 6 , —OR 6 , or —NR 6 R 7 ; 
 each X 9  is independently 
 
       
         
           
           
               
               
           
         
         each R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  is independently hydrogen or C 1 -C 6  alkyl; wherein one or more methylene groups of the C 1 -C 6  alkyl can be replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
       
       
         
           
           
               
               
           
         
         R 1  is hydrogen or C 1 -C 8  alkyl; 
         wherein optionally when proper any two of R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; and optionally X 2  and any one of R 2 , R 3 , and R 4  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; 
         A and B are each independently an optionally substituted monocyclic, bicyclic, or tricyclic aryl or heteroaryl; and 
         n is an integer from 2-4; each n 1  and n 2  is independently an integer from 0-2; n 3  is an integer from 1-2; n 4  is an integer from 1-3; and each one of n 9 , n 10 , n 11 , and n 12  is an integer from 0-1; and n 13  is an integer from 0-2, wherein when any one of the foregoing n integers is more than 1, the adjacent carbons represented by the integer can form a single or double bond. 
       
     
     
         63 . The method of  claim 62 , wherein the compound of Formula XXVII is selected from the group consisting of: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         64 . The method of  claim 18 , wherein the compound of Formula I′ is a compound of Formula XXXVII: 
       
         
           
           
               
               
           
         
         wherein 
         T is Tether; 
         R wh  is Warhead which is a radical resulting from the removal of a hydrogen of a compound of Formula I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, I-j, I-k, I-l, I-m, I-n, I-o, I-p, I-q, I-r, I-s, and I-t: 
       
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein
 each X 1  and X 8  is independently —O—, —S—, or —NR 6 —; 
 each X 2  is independently —R 6 , —OR 6 , or —NR 6 R 7 ; 
 each X 9  is independently 
 
       
         
           
           
               
               
           
         
         each R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  is independently hydrogen or C 1 -C 6  alkyl; wherein one or more methylene groups of the C 1 -C 6  alkyl can be replaced by —NR 1 —, —O—, —C(O)—, —S—, —SO—, —SO 2 —, or —C(═S)—; one or more methine groups of the C 1 -C 6  alkyl, when present, can be independently replaced by 
       
       
         
           
           
               
               
           
         
         R 1  is hydrogen or C 1 -C 8  alkyl; 
         wherein optionally when proper any two of R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; and optionally X 2  and any one of R 2 , R 3 , and R 4  when taken together form a 3- to 8-membered carbocyclic or heterocyclic ring or an aryl or heteroaryl group; 
         A and B are each independently an optionally substituted monocyclic, bicyclic, or tricyclic aryl or heteroaryl; and 
         n in Warhead is an integer from 2-4; each n 1  and n 2  is independently an integer from 0-2; n 3  is an integer from 1-2; n 4  is an integer from 1-3; and each one of n 9 , n 10 , n 11 , and n 12  is an integer from 0-1; and n 13  is an integer from 0-2, wherein when any one of the foregoing n integers is more than 1, the adjacent carbons represented by the integer can form a single or double bond; and 
         each n in Formula XXXVII is 0, 1, or 2. 
       
     
     
         65 . The method of  claim 64 , wherein the compound of Formula XXXVII is selected from the group consisting of: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         66 - 184 . (canceled)

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