Conjugate of a tubulysin analog with branched linkers

Abstract

Described is a conjugation of a tubulysin analog compound to a cell-binding molecule with branched/side-chain linkers for having better delivery of the conjugate compound and targeted treatment of abnormal cells. Also described are a branched-linkage method of conjugation of a tubulysin analog molecule to a cell-binding ligand, as well as methods of using the conjugate in targeted treatment of cancer, infection and autoimmune disease.

1 . A side chain-linkage compound of Formula (IV), which can readily react to a cell-binding molecule T to form a conjugate: 
       
         
           
           
               
               
           
         
         wherein 
         “ ” represents a single bond; 
         L 1  and L 2  are, the same or different, independently selected from O, NH, N, S, P, NNH, NHNH, N(R 3 ), N(R 3 )N(R 3 ′), CH, CO, C(O)NH, C(O)O, NHC(O)NH, NHC(O)O, polyethyleneoxy unit of formula (OCH 2 CH 2 ) p OR 3 , or (OCH 2 CH—(CH 3 )) p OR 3 , or NH(CH 2 CH 2 O) p R 3 , or NH(CH 2 CH(CH 3 )O) p R 3 , or N[(CH 2 CH 2 O) p R 3 ], [(CH 2 CH 2 O) p R 3 ′], or (OCH 2 CH 2 ) p COOR 3 , or CH 2 CH 2 (OCH 2 CH 2 ) p COOR 3 , wherein p and p′ are independently an integer selected from 0 to about 1000, or a combination of two or more thereof; C 1 -C 8  alkyl; C 2 -C 8  heteroalkyl, alkylcycloalkyl, or heterocycloalkyl; C 3 -C 8  aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl; or (Aa) r , r=1-12 (1 to 12 amino acid units), which is composed from natural or unnatural amino acids, or the same or different sequences of dipeptide, tripeptide, tetrapeptide, pentapeptide, hexapeptide, heptapeptide, octapeptide, nonapeptide, decapeptide, undecapeptide or dodecapeptide unit; 
         W is a stretcher unit having C 1 -C 18 , normally a self-immolative spacer, a peptidic unit, a hydrazone, a disulfide, a thioether, an ester, or an amide bond; w is 1 or 2 or 3; 
         V 1  and V 2  are independently a spacer unit and selected from O, NH, S, C 1 -C 8  alkyl, C 2 -C 8  heteroalkyl, alkenyl, or alkynyl, C 3 -C 8  aryl, heterocyclic, carbocyclic, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroaralkyl, heteroalkylcycloalkyl, or alkylcarbonyl, or (Aa) r , r=1-12 (1 to 12 amino acid units), which is composed from a natural or unnatural amino acid, or the same or different sequences of dipeptide, tripeptide, tetrapeptide, pentapeptide, hexapeptide, heptapeptide, octapeptide, nonapeptide, decapeptide, undecapeptide or dodecapeptide unit; or (CH 2 CH 2 O) p , p is 0-1000; and v 1  and v 2  are independently 0, 1 or 2, but v 1  and v 2  are not 0 at the same time; when v 1  or v 2  is 0, one of the side chain Q 1  or Q 2  fragment is absent; 
         Q 1  and Q 2  are independently represented by Formula (I-q1): 
       
       
         
           
           
               
               
           
         
         wherein   is a site linked to L 1  or L 2 ; G 1  and G 2  are independently OC(O), NHC(O), C(O), CH 2 , NH, OC(O)NH, NHC(O)NH, O, S, B, P(O)(OH), NHP(O)(OH), NHP(O)(OH)NH, CH 2 P(O)(OH)NH, OP(O)(OH)O, CH 2 P(O)(OH)O, NHS(O) 2 , NHS(O) 2 NH, CH 2 S(O) 2 NH, OS(O) 2 O, CH 2 S(O) 2 O, Ar, ArCH 2 , ArO, ArNH, ArS, ArNR 1 , or (Aa) q1 ; G 3  is OH, SH, OR 1 , SR 1 , OC(O)R 1 , NHC(O)R 1 , C(O)R 1 , CH 3 , NH 2 , NR 1 , +NH(R 1 ), +N(R 1 )(R 2 ), C(O)OH, C(O)NH 2 , NHC(O)NH 2 , BH 2 , BR 1 R 2 , P(O)(OH) 2 , NHP(O)(OH) 2 , NHP(O)(NH 2 ) 2 , S(O) 2 (OH), (CH 2 ) q1 C(O)OH, (CH 2 ) q1 P(O)(OH) 2 , C(O)(CH 2 ) q1 C(O)OH, OC(O)(CH 2 ) q1 C(O)OH, NHC(O)(CH 2 ) q1 C(O)OH, CO(CH 2 ) q1 P(O)(OH) 2 , NHC(O)O(CH 2 ) q1 C(O)OH, OC(O)NH(CH 2 ) q1 C(O)OH, NHCO(CH 2 ) q1 P(O)(OH) 2 , NHC(O)(NH)(CH 2 ) q1 C(O)OH, CONH(CH 2 ) q1 P(O)(OH) 2 , NHS(O) 2 (CH 2 ) q1 C(O)OH, CO(CH 2 ) q1 S(O) 2 (OH), NHS(O) 2 NH(CH 2 ) q1 C(O)OH, OS(O) 2 NH(CH 2 ) q1 C(O)OH, NHCO(CH 2 ) q1 S(O) 2 (OH), NHP(O)(OH)(NH)(CH 2 ) q1 C(O)OH, CONH(CH 2 ) q1 S(O)(OH), OP(O)(OH) 2 , (CH 2 ) q1 P(O)(NH) 2 , NHS(O) 2 (OH), NHS(O) 2 NH 2 , CH 2 S(O) 2 NH 2 , OS(O) 2 OH, OS(O) 2 OR 1 , CH 2 S(O) 2 OR 1 , Ar, ArR 1 , ArOH, ArNH 2 , ArSH, ArNHR 1 , or (Aa) q1 ; (Aa) q1  is a peptide containing the same or different sequence of natural or unnatural amino acids; X 1  and X 2  are independently O, CH 2 , S, S(O), NHNH, NH, N(R 1 ), +NH(R 1 ), +N(R 1 )(R 2 ), C(O), OC(O), OC(O)O, OC(O)NH, or NHC(O)NH; 
         Y 2  is O, NH, NR 1 , CH 2 , S, NHNH, or Ar; p 1 , p 2  and p 3  are independently 0-100 but are not 0 at the same time; q 1  and q 2  are independently 0-24; 
         R 1 , R 2 , R 3  and R 3 ′ are independently H, C 1 -C 8  alkyl; C 2 -C 8  heteroalkyl, or heterocyclic; C 3 -C 8  aryl, Ar-alkyl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, carbocyclic, or alkylcarbonyl; 
         alternatively, any one or more of W, Q 1 , Q 2 , L 1 , L 2 , V 1 , or V 2  can be independently absent but Q 1  and Q 2  are not absent at the same time; 
         D is tubulysin analog having following formula (II): 
       
       
         
           
           
               
               
           
         
         or a pharmaceutically acceptable salt, hydrate, or hydrated salt; or a polymorphic crystalline structure; or an optical isomer, racemate, diastereomer or enantiomer thereof, 
         wherein ----- is a linkage site that links to W independently; 
         wherein R 1 , R 2 , R 3 , and R 4  are independently H, C 1 -C 8  alkyl; C 2 -C 8  heteroalkyl, or heterocyclic; C 3 -C 8  aryl, Ar-alkyl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, carbocyclic, or alkylcarbonyl; or R 1  and R 2 , R 1  and R 3 , R 2  and R 3 , R 3  and R 4 , R 5  and R 6 , R 11  and R 12 , or R 13  and R 14  form a 3-7 membered carbocyclic, cycloalkyl, heterocyclic, heterocycloalkyl, aromatic or heteroaromatic ring system; R 1  and R 2  can be independently absent when they link to W independently or simultaneously, Y 1  is N or CH; 
         wherein R 5 , R 6 , R 8 , R 10  and R 11  are independently H, or C 1 -C 4  alkyl or heteroalkyl; 
         wherein R 7  is independently H, R 14 , —R 14 C(═O)X 1 R 15 ; or —R 14 X 1 R 15 ; X 1  is O, S, S—S, NH, CH 2  or NR 14 ; 
         wherein R 9  is selected from H, OH, —O—, ═O, —OR 14 , —OC(═O)R 14 , —OC(═O)NHR 14 —, —OC(═O)R 14 SSR 15 —, OP(═O)(OR 14 )—, —OC(═O)NR 14 R 15 , OP(═O)(OR 14 ), or OR 14 0P(═O)(OR 15 ); 
         wherein R 11  is independently H, R 14 , —R 14 C(═O)R 16 , —R 14 X 2 R 16 , —R 14 C(═O)X 2 , wherein X 2  is —O—, —S—, —NH—, —N(R 14 )—, —O—R 14 —, —S—R 14 —, —S(═O)—R 14 —, or —NHR 14 ; 
         wherein R 12  is R 15 , —OH, —SH, —NH 2 , NH, NHNH 2 , —NH(R 15 ), —OR 15 , —R 15 COR 16 , 
         R 15 COOR 16 , —R 15 C(O)NH 2 , —R 15 C(O)NHR 17 , —SR 16 , R 15 S(═O)R 16 , —R 15 P(═O)(OR 17 ) 2 , —R 15 OP(═O)(OR 17 ) 2 , —CH 2 OP(═O)(OR 17 ) 2 , —R 15 SO 2 R 17 , —R 15 X 2 R 16 , —R 15 C(═O)X 2 , where X 2  is —O—OH, SH, —S—, NH 2 , —NH—, —N(R 15 )—, —O—R 15 —, —S—R 15 —, —S(═O)—R 15 —, CH 2  or —NHR 5 —; 
         R 13  and R 14  are independently H, O, S, NH, N(R 15 ), NHNH, —OH, —SH, —NH 2 , NH, NHNH 2 , —NH(R 15 ), —OR 15 , CO, —COX 2 , —COX 2 R 16 , R 17 , F, Cl, Br, I, SR 16 , NR 16 R 17 , N═NR 16 , N═R 16 , NO 2 , SOR 16 R 17 , SO 2 R 16 , SO 3 R 16 , PR 16 , PR 16 R 17 , POR 16 R 17 , PO 2 R 16 R 17 , OP(O)(OR 17 ) 2 , OCH 2 OP(O)(OR 17 ) 2 , OC(O)R 17 , OC(O)OP(O)(OR 17 ) 2 , PO(OR 16 )(OR 17 ), OP(O)(OR 17 )OP(O)(OR 17 ) 2 , OC(O)NHR 17 , —O—(C 4 -C 12  glycoside), —N—(C 4 -C 12  glycoside); C 1 -C 8  alkyl or heteroalkyl; C 2 -C 8  alkenyl, alkynyl, heteroalkyl, or heterocycloalkyl; C 3 -C 8  aryl, Ar-alkyl, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl, or C 2 -C 8  ester, ether, or amide; or peptide containing 1-8 amino acids (NH(Aa) 1-8  or CO(Aa)i-s (N-terminal or C-terminal 1-8 same or different amino acids), or polyethyleneoxy unit of formula (OCH 2 CH 2 ) p  or (OCH 2 CH(CH 3 )) p , wherein p is an integer from 0 to about 1000, or a combination of two or more thereof; X 2  is O, S, S—S, NH, CH 2 , OH, SH, NH 2 , CHR 14  or NR 14 ; 
         R 15 , R 16  and R 17  are independently H, C 1 -C 8  alkyl or heteroalkyl; C 2 -C 8  alkenyl, alkynyl, heteroalkyl, or heterocycloalkyl; C 3 -C 8  aryl, Ar-alkyl, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl, or alkylcarbonyl, or Na + , K + , Cs + , Li + , Ca 2+ , Mg + , Zn 2+ , N + (R 1 )(R 2 )(R 3 )(R 4 ), or HN + (C 2 H 5 OH) 3  salt; 
         Y 1  and Y 2  are independently N or CH; q is 0 or 1; when q=0, Y 3  does not exist, Y 4 , Y 5 , Y 6  and Y 7  are independently CH, N, NH, O, S, or N(R′), thus Y 2 , Y 4 , YS, Y 6  and Y 7 form a heteroaromatic ring of furan, pyrrole thiophene, thiazole, oxazole, imidazole, pyrazole, triazole, tetrazole, or thiadiazole; when q=1, Y 3 , Y 4 , Y 5 , Y 6  and Y 7  are independently CH or N, thus Y 2 , Y 3 , Y 4 , Y 5 , Y 6  and Y 7  form an aromatic ring of benzene, pyridine, pyridazine, pyrimidine, pyrazine, triazine, tetrazine, or pentazine; 
         L V1  is a reacting group that can be reacted with a thiol, amine, carboxylic acid, selenol, phenol or hydroxyl group on a cell-binding molecule; L V1  is selected from OH; F; Cl; Br; I; nitrophenol; N-hydroxysuccinimide (NHS); phenol; dinitrophenol; pentafluorophenol; tetrafluorophenol; difluorophenol; mono-fluorophenol; pentachlorophenol; triflate; imidazole; dichlorophenol; tetrachlorophenol; 1-hydroxybenzotriazole; tosylate; mesylate; 2-ethyl-5-phenylisoxazolium-3′-sulfonate, anhydride formed by an acid itself, or formed with another anhydride; or an intermediate molecule generated with a condensation reagent for peptide coupling reactions, or for Mitsunobu reactions; the condensation reagent is selected from: EDC (N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide), DCC (dicyclohexyl-carbodiimide), N,N′-diisopropylcarbodiimide (DIC), N-cyclohexyl-N′-(2-morpholino-ethyl)carbodiimide metho-p-toluenesulfonate (CMC, or CME-CDI), 1,1′-carbonyldiimi-dazole (CDI), TBTU (0-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate), N,N,N′,N′-tetramethyl-O-(1H-benzo-triazol-1-yl)-uronium hexafluoro-phosphate (HBTU), (benzotriazol-1-yloxy)tris(dimethylamino)-phosphonium hexafluorophosphate (BOP), (benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyBOP), diethyl cyanophosphonate (DEPC), chloro-N,N,N′,N′-tetra-methylformamidiniumhexafluorophosphate, 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate (HATU), 1-[(dimethylamino)-(morpholino)methylene]-1H-[1,2,3]triazolo[4,5-b]pyridine-1-ium 3-oxide hexafluoro-phosphate (HDMA), 2-chloro-1,3-dimethyl-imidazolidinium hexafluorophosphate (CIP), chlorotripyrrolidinophosphonium hexafluorophosphate (PyCloP), fluoro-N,N,N′,N′-bis(tetramethylene)-formamidinium hexafluorophosphate (BTFFH), N,N,N′,N′-tetramethyl-S-(1-oxido-2-pyridyl)thiuronium hexafluorophosphate, O-(2-oxo-1(2H)pyridyl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (TPTU), S-(1-oxido-2-pyridyl)-N,N,N′,N′-tetramethylthiuronium tetrafluoroborate, O-[(ethoxycarbonyl)-cyanomethylenamino]-N,N,N′,N′-tetramethyluronium hexafluorophosphate (HOTU), (1-cyano-2-ethoxy-2-oxoethylidenaminooxy) dimethylamino-morpholino-carbenium hexafluorophosphate (COMU), O-(benzotriazol-1-yl)-N,N,N′,N′-bis(tetramethylene)-uronium hexafluorophosphate (HBPyU), N-benzyl-N′-cyclohexyl-carbodiimide (with, or without polymer-bound), dipyrrolidino(N-succinimidyl-oxy)carbenium hexafluoro-phosphate (HSPyU), chlorodipyrrolidinocarbenium hexafluorophosphate (PyClU), 2-chloro-1,3-dimethylimidazoli-dinium tetrafluoroborate(CIB), (benzotriazol-1-yloxy)dipiperidino-carbenium hexafluorophosphate (HBPipU), 0-(6-chlorobenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (TCTU), bromotris(dimethylamino)-phosphonium hexafluorophosphate (BroP), propylphosphonic anhydride (PPACA, T3P©), 2-morpholinoethyl isocyanide (MEI), N,N,N′,N′-tetramethyl-O—(N-succinimidyl)uronium hexafluorophosphate (HSTU), 2-bromo-1-ethyl-pyridinium tetrafluoroborate (BEP), O-[(ethoxycarbonyl)cyano-methylenamino]-N,N,N′,N′-tetramethyluronium tetrafluoroborate (TOTU), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholiniumchloride (MMTM, DMTMM), N,N,N′,N′-tetramethyl-O—(N-succinimidyl)uronium tetrafluoroborate (TSTU), O-(3,4-dihydro-4-oxo-1,2,3-benzotriazin-3-yl)-N,N,N′,N′-tetramethyluronium tetrafluoro-borate (TDBTU),1,1′-(azodicarbonyl)-dipiperidine (ADD), di-(4-chlorobenzyl)azodicarboxylate (DCAD), di-tert-butyl azodicarboxylate (DBAD), diisopropyl azodicarboxylate (DIAD), diethyl azodicarboxylate (DEAD); L V1  is also an anhydride, formed by an acid itself or formed with another C 1 -C 8  acid anhydride. 
       
     
     
         2 . The side chain-linkage compound according to  claim 1 , wherein L V1  is selected from: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         wherein X 1 ′ is F, Cl, Br, I or L v3 ; X 2 ′ is O, NH, N(R 1 ), or CH 2 ; R 3  is independently H, aromatic, heteroaromatic, or aromatic group wherein one or several H atoms are replaced independently by —R 1 ,-halogen, —OR 1 , —SR 1 , —NR 1 R 2 , — NO 2 , —S(O)R 1 , —S(O) 2 R 1 , or —COOR 1 ; L v3  is a leaving group selected from F, Cl, Br, I, nitrophenol; N-hydroxysuccinimide (NHS); phenol; dinitrophenol; pentafluorophenol; tetrafluorophenol; difluorophenol; monofluorophenol; pentachlorophenol; triflate; imidazole; dichlorophenol; tetrachlorophenol; 1-hydroxybenzotriazole; tosylate; mesylate; 2-ethyl-5-phenylisoxazolium-3′-sulfonate, anhydride formed by an acid itself, or formed with another anhydride: acetyl anhydride, formyl anhydride; or an intermediate molecule generated with a condensation reagent for peptide coupling reactions or for Mitsunobu reactions. 
       
     
     
         3 . The side chain-linkage compound according to  claim 1 , wherein Q 1  and Q 2  are independently a C 2 -C 100  polycarboxylacid; a C 2 -C 100  polyalkylamine; a C 6 -C 100  oligosaccharide or polysaccharide; a C 6 -C 100  zwitterionic betaine or zwitterionic poly(sulfobetaine)) (PSB) that consists of a quaternary ammonium cation and a sulfonate anion; a C 6 -C 100  biodegradable polymer composed of poly(lactic/glycolic acid) (PLGA), poly(acrylates), chitosan, copolymer of N-(2-hydroxypropyl)methacrylamide, poly[2-(methacryloyloxy)ethyl phosphorylcholine](PMPC), poly-L-glutamic acid, poly(lactide-co-glycolide) (PLG), poly(lactide-co-glycolide), poly(ethylene glycol) (PEG), poly(propylene glycol) (PPG), poly(lactide-co-glycolide), poly(ethylene glycol)-modified peptide, poly(ethylene glycol)-containing an amino acid or peptide, poly(ethylene glycol)-modified lipid, poly(ethylene glycol)-modified alkylcarboxic acid, poly(ethylene glycol)-modified alkylamine, poly(lactide-co-glycolide, hyaluronic acid (HA) (glycosaminoglycan), heparin/heparan sulfate (HSGAGs), chondroitin sulfate/dermatan sulfate (CSGAGs), poly(ethylene glycol)-modified alkylsulfate, poly(ethylene glycol)-modified alkylphosphate, or poly(ethylene glycol)-modified alkyl quaternary ammonium. 
     
     
         4 . The side chain-linkaged compound according to  claim 1 , wherein Q 1  and Q 2  are independently selected from Iq-01 to Iq-35: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         wherein R 25  and R 25 ′ are independently selected from H; HC(O), CH 3 C(O), CH 3 C(NH), C 1 -C 18  alkyl, C 1 -C 18  alkyl, alkyl-Y 1 —SO 3 H, C 1 -C 18  alkyl-Y 1 —PO 3 H 2 , C 1 -C 18  alkyl-Y 1 —CO 2 H, C 1 -C 18  alkyl-Y 1 —N + R 1 ′R 2 ′R 3 ′R 4 ′, C 1 -C 18  alkyl-Y 1 —CONH 2 , C 2 -C 18  alkylene, C 2 -C 18  ester, C 2 -C 18  ether, C 2 -C 18  amine, C 2 -C 18  alkyl carboxylamide, C 3 -C 18  Aryl, C 3 -C 18  cyclic alkyl, C 3 -C 18  heterocyclic, 1-24 amino acids; C 2 -C 18  lipid, a C 2 -C 18  fatty acid or a C 2 -C 18  fatty ammonium lipid; X 1  and X 2  are independently selected from NH, N(R 1 ′), O, CH 2 , S, C(O), S(O), S(O 2 ), P(O)(OH), NHNH, CH═CH, Ar or (Aa) q1 , q 1 =0-24 (0-24 amino acids, q 1 =0 means absent); X 1 , X 2 , X 3 , X 4 , Y 1 , Y 2  and Y 3  are independently selected from NH, N(R 1 ′), O, C(O), CH 2 , S, S(O), NHNH, C(O), OC(O), OC(O)O, OC(O)NH, NHC(O)NH, Ar or (Aa) q1 , X 1 , X 2 , X 3 , X 4 , Y 1 , Y 2  and Y 3  can be independently absent; p 1 , p 2  and p 3  are independently 0-100 but are not 0 at the same time; q 1 , q 2  and q 3  are independently 0-24; R 1 ′, R 2 ′, R 3 ′ and R 4 ′ are independently selected from H and C 1 -C 6  alkyl; Aa is natural or unnatural amino acid; Ar or (Aa) q1 , is the same or different sequence of peptides; q 1 =0 means (Aa) q1  absent. 
       
     
     
         5 . The side chain-linkaged compound according to  claim 1 , wherein D (tubulysin structure) is selected from 1-01 to 1-75: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         or their pharmaceutically acceptable salts, hydrates, or hydrated salts; or polymorphic crystalline structures of these compounds; or their optical isomers, racemates, diastereomers or enantiomers; wherein R 20  is H; C 1 -C 8  linear or branched alkyl or heteroalkyl; C 2 -C 8  linear or branched alkenyl, alkynyl, alkylcycloalkyl, or heterocycloalkyl; C 3 -C 8  linear or branched aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl; carbonate (—C(O)OR 17 ), carbamate (—C(O)NR 17 R 18 ); or C 1 -C 8  carboxylate, ester, ether, or amide; or 1-8 amino acids; or polyethyleneoxy unit of formula (OCH 2 CH 2 ) p  or (OCH 2 CH(CH 3 )) p , wherein p is an integer from 0 to about 1000; or R 20  is absent and the oxygen atom forms a ketone, or a combination of two of more thereof, Z 2  and Z 3  are independently H, OH, NH 2 , O, NH, COOH, COO, C(O), C(O), C(O)NH, C(O)NH 2 , R 18 , OCH 2 OP(O)(OR 1 8) 2 , OC(O)OP(O)(OR 1 8) 2 , OPO(OR 1 8) 2 , NHPO(OR 1 8) 2 , OP(O)(OR 1 8)OP(O)(OR 1 8) 2 , OC(O)R 1 8, OC(O)NHR 1 8, OSO 2 (OR 1 8), O—(C 4 -C 12 -glycoside), C 1 -C 8  linear or branched alkyl or heteroalkyl; C 2 -C 8  linear or branched alkenyl, alkynyl, alkylcycloalkyl, or heterocycloalkyl; C 3 -C 8  linear or branched aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl; carbonate (—C(O)OR 17 ), carbamate (—C(O)NR 17 R 18 ); R 17  and R 18  are independently H, C 1 -C 8  linear or branched alkyl or heteroalkyl; C 2 -C 8  linear or branched alkenyl, alkynyl, alkylcycloalkyl, or heterocycloalkyl; C 3 -C 8  linear or branched aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl; carbonate (—C(O)OR 17 ), carbamate (—C(O)NR 17 R 18 ); R 19  is H, OH, NH 2 , OSO 2 (OR 18 ), XCH 2 OP(O)(OR 18 ) 2 , XPO(OR 18 ) 2 , XC(O)OP(O)(OR 18 ) 2 , XC(O)R 18 , XC(O)NHR 18 , C 1 -C 8  alkyl or carboxylate; C 2 -C 8  alkenyl, alkynyl, alkylcycloalkyl, or heterocycloalkyl; C 3 -C 5  aryl or alkylcarbonyl; or pharmaceutical salts; X is O, S, NH, NHNH, or CH 2 ; R 7  is defined the same as in  claim 1 . 
       
     
     
         6 . The side chain-linkaged compound according to  claim 1 , wherein W, L 1 , L 2 , V 1 , and V 2  independently is composed of one or more linker components of the following structures: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       or L- or D-, natural or unnatural peptides containing 1-20 the same or different amino acids; 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         wherein   is a site of linkage; X 2 , X 3 , X 4 , X 5 , and X 6  are independently selected from NH; NHNH; N(R 3 ); N(R 3 )N(R 3 ′); 0; S; C 1 -C 6  alkyl; C 2 -C 6  heteroalkyl, alkylcycloalkyl, or heterocycloalkyl; C 3 -C 8  aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl; or 1-8 amino acids; wherein R 3  and R 3 ′ are independently H; C 1 -C 8  alkyl; C 2 -C 8  hetero-alkyl, alkylcycloalkyl, or heterocycloalkyl; C 3 -C 8  aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl; or C 1 -C 8  ester, ether, or amide; or polyethyleneoxy unit of formula (OCH 2 CH 2 ) p  or (OCH 2 CH(CH 3 )) p , wherein p is an integer from 0 to about 1000, or a combination of two or more thereof. 
       
     
     
         7 . The side chain-linkaged conjugate compound according to  claim 1 , wherein W, L 1 , L 2 , V 1 , and V 2  independently is composed of:
 (A): a self-immolative component, peptidic units, a hydrazone bond, a disulfide, an ester, an oxime, an amide, or a thioether bond; the self-immolative unit includes aromatic compounds that are electronically similar to para-aminobenzyl-carbamoyl (PAB) groups, 2-aminoimidazol-5-methanol derivatives, heterocyclic PAB analogs, beta-glucuronide, and ortho or para-aminobenzylacetals; or one of the following structures:   
       
         
           
           
               
               
           
         
         wherein the (*) atom is a point of attachment of another component; X 1 , Y 1 , Z 2  and Z 3  are independently NH, O, or S; Z 1  is independently H, NHR 1 , OR 1 , SR 1 , or COX 1 R 1 , wherein X 1  and Ri are defined the same as in  claim 1 ; v is 0 or 1; U 1  is independently H, OH, C 1 -C 6  alkyl, (OCH 2 CH 2 ) n , F, Cl, Br, I, OR 5 , SR 5 , NR 5 R 5 ′, N═NRs, N═R 5 , NR 5 R 5 ′, NO 2 , SOR 5 R 5 ′, SO 2 R 5 , SO 3 R 5 , OSO 3 R 5 , PR 5 R 5 ′, POR 5 R 5 ′, PO 2 R 5 R 5 ′, OPO(OR 5 )(OR 5 ′), or OCH 2 PO(OR 5 (OR 5 ′), wherein R 5  and R 5 ′ are independently selected from H, C 1 -C 8  alkyl; C 2 -C 8  alkenyl, alkynyl, heteroalkyl, or amino acid; C 3 -C 8  aryl, heterocyclic, carbocyclic, cycloalkyl, heterocycloalkyl, heteroaralkyl, alkylcarbonyl, or glycoside; or pharmaceutical cation salts thereof; 
         (B): a non-self-immolative linker component containing one of the following structures: 
       
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         wherein the (*) atom is a site of attachment; X 1 , Y 1 , U 1 , R 5 , R 5 ′ are defined as above; r is 0-100; m and n are 0-20 independently; 
         (C): a releasable component that at least one bond that can be broken under physiological conditions: a pH-labile, acid-labile, base-labile, oxidatively labile, metabolically labile, biochemically labile or enzyme-labile bond, which having one of the following structures: 
         —(CR 5 R 6 ) m (Aa) r (CR 7 R 8 ) n (OCH 2 CH 2 ) t —, —(CR 5 R 6 ) m (CR 7 R 8 ) n (Aa) r (OCH 2 CH 2 ) t —, —(Aa) r —(CR 5 R 6 ) m (CR 7 R 8 ) n (OCH 2 CH 2 ) t —, —(CR 5 R 6 ) m (CR 7 R 8 ) n (OCH 2 CH 2 ) r (Aa) t —, —(CR 5 R 6 ) m —(CR 7 ═CR 5 )(CR 9 R 10 ) n (Aa) t (OCH 2 CH 2 ) r —, —(CR 5 R 6 ) m (NR 11 CO)(Aa) t (CR 9 R 10 ) n —(OCH 2 CH 2 ) r —(CR 5 R 6 ) m (Aa) t (NR 11 CO)(CR 9 R 10 ) n (OCH 2 CH 2 ) r —, —(CR 5 R 6 ) m (OCO)(Aa) t (CR 9 R 10 ) n —(OCH 2 CH 2 ) r —, —(CR 5 R 6 ) m (OCNR 7 )(Aa) t (CR 9 R 10 ) n (OCH 2 CH 2 ) r —, —(CR 5 R 6 ) m (CO)(Aa) t —(CR 9 R 10 ) n (OCH 2 CH 2 ) r —(CR 5 R 6 ) m (NR 11 CO)(Aa) t (CR 9 R 10 ) n (OCH 2 CH 2 ) r —, —(CR 5 R 6 ) m —(OCO)(Aa) t (CR 9 R 10 ) n —(OCH 2 CH 2 ) r —, —(CR 5 R 6 ) m (OCNR 7 )(Aa) t (CR 9 R 10 ) n (OCH 2 CH 2 ) r —, —(CR 5 R 6 ) m (CO)(Aa) t (CR 9 R 10 ) n —(OCH 2 CH 2 ) r —, —(CR 5 R 6 ) m -phenyl-CO(Aa) t (CR 7 R 8 ) n —, —(CR 5 R 6 ) m -furyl-CO(Aa) t (CR 7 R 8 ) n —, —(CR 5 R 6 ) m —oxazolyl-CO(Aa) t (CR 7 R 8 ) n —, —(CR 5 R 6 ) m -thiazolyl-CO(Aa) t (CCR 7 R 8 ) n —, —(CR 5 R 6 ) t -thienyl-CO(CR 7 R 8 ) n —, —(CR 5 R 6 ) t -inidazolyl-CO—(CR 7 R 8 ) n —, —(CR 5 R 6 ) t -morpholino-CO(Aa) t —(CR 7 R 8 ) n —, —(CR 5 R 6 ) t piperazino-CO(Aa) t —(CR 7 R 8 ) n —, —(CR 5 R 6 ) t —N-methylpiperazin-CO(Aa) t —(CR 7 R 8 ) n —, —(CR 5 R) m —(Aa) t phenyl-, —(CR 5 R 6 ) m —(Aa) t furyl-, —(CR 5 R 6 ) m -oxazolyl(Aa) r —, —(CR 5 R 6 ) m -thiazolyl(Aa) t —, —(CR 5 R 6 ) m -thienyl-(Aa) t —, —(CR 5 R( ) m -imidazolyl(Aa) t —, —(CR 5 R 6 ) m -morpholino-(Aa) t —, —(CR 5 R 6 ) m -piperazino-(Aa) t —, —(CR 5 R 6 ) m —N-methylpiperazino-(Aa) t —, —K(CR 5 R 6 ) m (Aa) r (CR 7 R 8 ) n (OCH 2 CH 2 ) t —, —K(CR 5 R 6 ) m (CR 7 R 8 ) n —(Aa) r (OCH 2 CH 2 ) t —, —K(Aa) r —(CR 5 R 6 ) m (CR 7 R 8 ) n (OCH 2 CH 2 ) t —, —K(CR 5 R 6 ) m —(CR 7 R 8 ) n (OCH 2 —CH 2 ) r (Aa) t —, —K(CR 5 R 6 ) m —(CR 7 ═CR 8 )(CR 9 R 10 ) n (Aa) t (OCH 2 CH 2 ) r —, —K(CR 5 R 6 ) m —(NR 11 CO—)(Aa) t (CR 9 R 10 ) n (OCH 2 C 12 ) r —, —K(CR 5 R 6 ) m (Aa) t (NR 11 CO)(CR 9 R 10 ) n (OCH 2 CH 2 ) r —, —K(CR 5 R 6 ) m (OCO)(Aa) t (CR 9 R 10 ) n —(OCH 2 CH 2 ) r —, —K(CR 5 R 6 ) m (OCNR 7 )(Aa) t (CR 9 R 10 ) n —(OCH 2 CH 2 ) r —, —K(CR 5 R 6 ) m (CO)(Aa) t —(CR 9 R 10 ) n (OCH 2 CH 2 ) r —, —K(CR 5 R 6 ) m (NR 11 CO)(Aa) t —(CR 9 R 10 ) n (OCH 2 CH 2 ) r —, —K(CR 5 R 6 ) m —(OCO)(Aa) t (CR 5 R 10 ) n (OCH 2 CH 2 ) r —, —K(CR 5 R 6 ) m —(OCNR 7 )(Aa) t (CR 9 R 10 ) n (OCH 2 CH 2 ) r —, —K—(CR 5 R 6 ) m (CO)(Aa) t (CR 9 R 10 ) n (OCH 2 CH 2 ) r —, —K(CR 5 R 6 ) m -phenyl—CO(Aa) t (CR 7 R 8 ) n —, —K—(CR 5 R 6 ) m -furyl-CO(Aa) t —(CR 7 R 8 ) n —, —K(CR 5 R 6 ) m -oxazolyl-CO(Aa) t —(CR 7 R 8 ) n —, —K(CR 5 R 6 ) m - thiazolyl-CO(Aa) t —(CR 7 R 8 ) n —, —K(CR 5 R 6 ) t -thienyl-CO(CR 5 R 8 ) n —, —K(CR 5 R 6 ) t imidazolyl-CO—(CR 7 R 8 ) n —, —K(CR 5 R 6 ) t -morpholino-CO—(Aa) t (CR 7 R 8 ) n —, —K(CR 5 R 6 ) t -piperazino-CO(Aa) t —(CR 7 R 8 ) n —, —K(CR 5 R 6 ) t —N-methylpiperazin-CO(Aa) t (CR 7 R 8 ) n —, —K(CR 5 R) m —(Aa) t phenyl, —K—(CR 5 R 6 ) m —(Aa) t furyl-, —K(CR 5 R 6 ) m -oxazolyl(Aa) t —, —K(CR 5 R 6 ) m -thiazolyl(Aa) t —, —K(CR 5 R 6 ) m -thienyl-(Aa) t —, —K(CR 5 R 6 ) m -imidazolyl(Aa) t —, —K(CR 5 R 6 ) m -morpholino(Aa) t —, —K(CR 5 R 6 ) m -piperazino-(Aa) t —, —K(CR 5 R 6 ) m —N-methylpiperazino(Aa) t —; wherein Aa, m, and n are described above; t and r are 0-100 independently; R 3 , R 4 , R 5 , R 6 , R 7 , and R 8  are independently chosen from H; halide; C 1 -C 8  alkyl; C 2 -C 8  aryl, alkenyl, alkynyl, ether, ester, amine or amide, which is optionally substituted by one or more halide, CN, NR 1 R 2 , CF 3 , OR 1 , Aryl, heterocycle, S(O)R 1 , SO 2 R 1 , —CO 2 H, —SO 3 H, —OR 1 , —CO 2 R 1 , —CONR 1 , —PO 2 R 1 R 2 , —PO 3 H or P(O)R 1 R 2 R 3 ; K is NR 1 , —SS—, —C(═O)—, —C(═O)NH—, —C(═O)O—, —C═NH—O—, —C═N—NH—, —C(═O)NH—NH—, O, S, Se, B, Het (heterocyclic or heteroaromatic ring having C 3 -C 8 ), or a peptide containing 1-20 amino acids. 
       
     
     
         8 . The side chain-linkaged compound according to  claim 1  having one of the following structures: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         or their pharmaceutically acceptable salts, hydrates, or hydrated salts; or polymorphic crystalline structures of these compounds; or their optical isomers, racemates, diastereomers or enantiomers: wherein X 1 , X 2 , and X 3  are independently O, S, NH, NHNH, or CH 2 ; Z 2  and Z 3  are independently NH, O, or S; p, p 1 , p 2 , and p 3  are independently 1-100; q 1  and q 2  are independently selected from 0-24; L v3  is a leaving group selected from F, Cl, Br, I, nitrophenol; N-hydroxysuccinimide (NHS); phenol; dinitrophenol; pentafluorophenol; tetrafluorophenol; difluorophenol; monofluorophenol; pentachlorophenol; triflate; imidazole; dichlorophenol; tetrachlorophenol; 1-hydroxy benzotriazole; tosylate; mesylate; 2-ethyl-5-phenylisoxazolium-3′ sulfonate, anhydride formed by an acid itself, or formed with another anhydride: acetyl anhydride, formyl anhydride; or an intermediate molecule generated with a condensation reagent for peptide coupling reactions or for Mitsunobu reactions; Aa is natural or unnatural amino acid; r is 0-12; (Aa) r  is a peptide containing the same or different sequence of amino acids when r>2; r=0 means (Aa) r  absent; R 25  and R 25 ′ are independently selected from H; HC(O), CH 3 C(O), CH 3 C(NH), C 1 -C 18  alkyl, C 1 -C 18  alkyl, alkyl-Y 1 —SO 3 H, C 1 -C 18  alkyl-Y 1 —PO 3 H 2 , C 1 -C 18  alkyl-Y 1 —CO 2 H, C 1 -C 18  alkyl-Y 1 —N + R 1 ′R 2 ′R 3 ′R 4 ′, C 1 -C 18  alkyl-Y 1 —CONH 2 , C 2 -C 18  alkylene, C 2 -C 18  ester, C 2 -C 18  ether, C 2 -C 18  amine, C 2 -C 18  alkyl carboxylamide, C 3 -C 18  Aryl, C 3 -C 18  cyclic alkyl, C 3 -C 18  heterocyclic, 1-24 amino acids; C 2 -C 18  lipid, a C 2 -C 18  fatty acid or a C 2 -C 18  fatty ammonium lipid; and m is 0-20.