Conjugates of tubulysin analogues with branched linkers

Abstract

To provide 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, and also to provide 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.SOLUTION: Provided is a side chain-linkaged conjugate compound of Formula (III): in the formula, T is a cell-binding agent selected from the group consisting of an antibody, a single chain antibody, an antibody fragment that binds to a target cell, a monoclonal antibody, a single chain monoclonal antibody, a monoclonal antibody fragment that binds to a target cell, a chimeric antibody, a chimeric antibody fragment that binds to the target cell, a domain antibody, a domain antibody fragment that binds to the target cell, an adnectin that mimics an antibody, DARPins, a lymphokine, a hormone, a vitamin, a growth factor, a colony stimulating factor, a nutrient-transport molecule (a transferrin), and / or a cell-binding peptide, etc.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention provides a method for the delivery of conjugated compounds with better pharmacokinetics and much more precise differential Cell-binding molecules and chimeric molecules with branched linkages (side chains) for targeted therapy of normal cells. The present invention also relates to conjugates with tubulysin analog compounds. Branched binding methods for molecular cleavage to cell-binding ligands and the treatment of cancer, infectious diseases, and autoimmune diseases The present invention relates to methods of using the conjugates in targeted therapy. [Background technology]

[0002] Antibody-drug conjugates (ADCs) are being investigated for the treatment of relapsed / refractory Hodgkin lymphoma with brentuximab. Buvedotin (Adcetris) (Non-patent Documents 1 and 2) and Adcetris for Recurrent HER2+ Breast Cancer As evidenced by the clinical success of rastuzumab emtansine (Non-patent Documents 3 and 4), It has become one of the most promising targeted therapies for cancer. It has three key components: monoclonal antibodies, cytotoxic In addition to the conjugates of the active payload and conditional ADC, All of these binding sites are important factors for the success of ADCs (Non-Patent Documents 5 and 6) It took 30 years to study each component of the ADC. However, the conjugated Drugs must contain specific reactive functional groups to ensure circulation stability and to interact with antigens. The drug is easily released upon binding to the carboxyl group and cellular uptake, and once the carboxyl group binds to the carboxyl group and the payload is released, the drug is released into the circulation. It is important to ensure that the dopamine components do not off-target and damage normal tissues. Therefore, the scope of conjugate technology remains limited (Non-Patent Documents 7-10).

[0003] In the early days of ADCs, especially those targeting liquid tumors, the conjugates used were very are unstable, leading to the release of free drug in the circulation and consequent off-target toxicity In the current generation of ADCs, the conjugates are more stable and have low cytotoxicity (Non-Patent Document 11). These drugs are highly potent (Non-Patent Document 12). However, off-target toxicity has been limited to the following: This remains a major challenge in the development of ADC drugs (Non-Patent Document 13). In clinical practice, ado-trastuzumab uses a stable (non-cleavable) MCC conjugate. Emtansine (T-DM1, Kadcyla®) is a treatment for HER2-positive metastatic breast cancer. Patients with metastatic breast cancer (mBC) or who have already been treated for mBC or who are receiving adjuvant therapy showed a significant benefit to patients who developed HER2 tumor recurrence within 6 months of receiving HER2 therapy (Non-Patent Literature 14-16) However, when comparing efficacy and secondary toxicity, T-DM1 The benefit for patients with HER2-positive unresectable locally advanced or metastatic As first-line treatment for patients with breast cancer and second-line treatment for HER2-positive advanced gastric cancer Clinical trials as a therapeutic agent have failed (Non-Patent Documents 17-20).

[0004] Research and development of ADC chemistry and design to address the issue of off-target toxicity Therefore, currently the range of linker-payload moieties and conjugate chemistries is limited to just potential payloads. Beyond the standard, specifically addressing the activity of ADC conjugate-payloads towards targets / diseases Currently, many drug developers and academic researchers The institution is looking to improve batch-to-batch consistency in ADC production as well as develop compounds with longer circulating half-lives and efficacy. High efficacy, potentially reduced off-target toxicity, and narrow range in vivo pharmacokinetics of ADCs Novel, reliable, specific conjugates and ADC moieties with potential PK properties We have focused on establishing methods for specific conjugation (Non-Patent Documents 23-27). These specific conjugation methods reported in

[28] include the use of designed cysteines ( 29, Patent Documents 1-5), selenocysteine ​​(Non-Patent Documents 30-31, Patent Document 6), Stain-containing tags and perfluoroaromatic reagents (Non-Patent Document 32), thiol fucose (non- Patent Document 33), unnatural amino acids (Non-Patent Documents 34 to 37, Patent Documents 7 to 17), dibromo Conjugation of momaremides to reduced intermolecular disulfides by re-crosslinking (Non-Patent Document 38), Bis-sulfone reagents (Non-patent Document 39, Patent Documents 18-19), dibromopyridazinedione (Non-patent Document 40), galactosyl- and sialyltransferases (Non-patent Document 41, Patent Document 20), formylglycine synthase (FGE) (Non-patent Document 42, Patent Document 21 ~25), phosphopantetheinyl transferases (PPTases) (Non-patent Document 4 3), sortase A (Non-patent document 44), genetically introduced glutamine tag and Stre Ptoverticillium mobaraense transglutaminase (mT G) (Non-Patent Documents 45-46, Patent Document 26) or microbial transglutaminase (MT Gase) (Non-patent documents 47-48, Patent documents 27-28), Isopeptide bond-forming enzymes / bacteria that form peptide bonds (Non-patent documents 49-51) This includes the incorporation of

[0005] We have developed bromomaleimide and dibromomaleimide conjugates (Patent Document 29), 2,3- Disubstituted succinic acid / 2-monosubstituted / 2,3-disubstituted fumaric or maleic acid linkers (Patent Document 30 and 31), acetylenedicarboxylic acid conjugates (Patent Documents 32 and 33), or hydra The reduced interchain disulfide bonds of natural antibodies can be reversed, for example, by using a diamine conjugate (Patent Document 34). Several conjugation methods have been disclosed for re-crosslinking the thiol pairs in the hydroxyl groups. The prepared ADCs are more selective than conventional non-selective conjugates via cysteine ​​or lysine residues of antibodies. Here, we demonstrate a therapeutic index window superior to that of the previous study. The invention of a tubulysin conjugate is disclosed. The long side chain linker allows the antibody-drug complex to hydrolyze. It prevents hydrolysis by enzymes such as proteinases or esterases, making it more readily available in the circulation. leading to stable conjugates.

[0006] Tubulysins as potent cytotoxic agents are well known in the art and can be synthesized by known methods. As such, they can be isolated from natural sources or prepared synthetically according to known methods (e.g., For example, non-patent literature 52-69, patent applications: patent literature 35-66). The construction of tubulysin conjugates for targeted treatment of infectious and autoimmune diseases has been disclosed (Patent 67) The tubulysin conjugates of the present invention containing long branched (side chain) linkers are useful for targeted delivery. Extend the half-life of the conjugate during delivery and minimize exposure to non-target cells, tissues, or organs in the bloodstream. This minimizes the risk of off-target toxicity, resulting in lower off-target toxicity. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] U.S. Patent No. 8,309,300 [Patent Document 2] U.S. Patent No. 7,855,275 [Patent Document 3] U.S. Patent No. 7,521,541 [Patent Document 4] U.S. Patent No. 7,723,485 [Patent Document 5] International Publication No. WO2008 / 141044 [Patent Document 6] U.S. Patent No. 8,916,159 [Patent Document 7] U.S. Patent No. 8,778,631 [Patent Document 8] U.S. Patent Application No. 20100184135 [Patent Document 9] International Publication No. WO2010 / 081110 [Patent Document 10] International Publication No. WO2006 / 069246 [Patent Document 11] International Publication No. 2007 / 059312 [Patent Document 12] U.S. Patent No. 7,332,571 [Patent Document 13] U.S. Patent No. 7,696,312 [Patent Document 14] U.S. Patent No. 7,638,299 [Patent Document 15] International Publication No. WO2007 / 130453 [Patent Document 16] U.S. Patent No. 7,632,492 [Patent Document 17] U.S. Patent No. 7,829,659 [Patent Document 18] International Publication No. WO2013 / 190272 [Patent Document 19] International Publication No. WO2014 / 064424 [Patent Document 20] U.S. Patent Application No. 20140294867 [Patent Document 21] U.S. Patent No. 7,985,783 [Patent Document 22] U.S. Patent No. 8,097,701 [Patent Document 23] U.S. Patent No. 8,349,910 [Patent Document 24] U.S. Patent Application No. 20140141025 [Patent Document 25] U.S. Patent Application No. 20100210543 [Patent Document 26] U.S. Patent No. 8,871,908 [Patent Document 27] U.S. Patent Application No. 20130189287 [Patent Document 28] U.S. Patent No. 7,893,019 [Patent Document 29] International Publication No. WO2014 / 009774 [Patent Document 30] International Publication No. WO2015 / 155753 [Patent Document 31] International Publication No. WO2016 / 0596228 [Patent Document 32] International Publication No. WO2015 / 151080 [Patent Document 33] International Publication No. WO2016 / 0596228 [Patent Document 34] International Publication No. WO2015 / 151081 [Patent Document 35] Canadian Patent Publication No. CA2710693 (2011) [Patent Document 36] European Patent Application No. 2174947 (2010) [Patent Document 37] International Publication No. WO2010 / 034724 [Patent Document 38] International Publication No. WO2010 / 033733 [Patent Document 39] International Publication No. WO2009 / 002993 [Patent Document 40] International Publication No. WO2009 / 134279 [Patent Document 41] International Publication No. WO2009 / 012958 [Patent Document 42] U.S. Patent Application No. 20110263650 [Patent Document 43] U.S. Patent Application No. 20110021568 [Patent Document 44] International Publication No. WO2009 / 095447 [Patent Document 45] International Publication No. WO2009 / 055562 [Patent Document 46] International Publication No. WO2008 / 112873 [Patent Document 47] International Publication No. WO2009 / 026177 [Patent Document 48] International Publication No. WO2008 / 138561 [Patent Document 49] International Publication No. WO2008 / 125116 [Patent Document 50] International Publication No. WO2008 / 076333 [Patent Document 51] U.S. Patent Application No. 20070041901 [Patent Document 52] International Publication No. WO2006 / 096754 [Patent Document 53] International Publication No. WO2006 / 056464 [Patent Document 54] International Publication No. WO2006 / 033913 [Patent Document 55] German Patent Publication No. 102004030227 [Patent Document 56] International Publication No. WO2004 / 005327 [Patent Document 57] International Publication No. WO2004 / 005326 [Patent Document 58] International Publication No. WO2004 / 005269 [Patent Document 59] U.S. Patent Application No. 20040249130 [Patent Document 60] German Patent Publication No. 10254439 [Patent Document 61] German Patent Publication No. 10241152 [Patent Document 62] German Patent Publication No. 10008089 [Patent Document 63] International Publication No. WO2002 / 077036 [Patent Document 64] German Patent Publication No. 19638870 [Patent Document 65] U.S. Patent Publication No. 20120129779 [Patent Document 66] U.S. Patent Application No. 20110027274 [Patent Document 67] PCT / IB2012 / 053554 issue [Non-patent literature]

[0008] [Non-Patent Document 1] Hematol Oncol. Clin. North. Am, 2014, 28, 13-25 [Non-patent document 2] Blood 2015, 125, 1236-43 [Non-patent document 3] Ther. Adv. Med. Oncol. 2014, 6(5), 202-9 [Non-patent document 4] J. Med. Chem. 2014, 57, 6949-64 [Non-patent document 5] Bioconjugate Chem., 2010, 21, 5-13 [Non-patent document 6] Biologicals 2015, 43, 318-32

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[0009] The present invention relates to a tubulysin analogue Cell-binding agents / molecules It also provides branched bonds to tubulysin analogs with side chain linkers. Cell-binding agents / molecules Also provides a method for combining with 。 [Means for solving the problem]

[0010] In one embodiment of the invention, the conjugate containing the side chain linkage is represented by formula (I): [ka]

[0011] During the ceremony, [ka] represents a single bond; n is 1 to 30;

[0012] T is an antibody, a single-chain antibody, an antibody fragment that binds to a target cell, a monoclonal antibody, a single-chain monoclonal antibody Clonal antibodies, monoclonal antibody fragments that bind to target cells, chimeric antibodies, and antibodies that bind to target cells chimeric antibody fragments, domain antibodies, domain antibody fragments, and antibodies that bind to target cells Mimicking Adnectins, DARPins, lymphokines, hormones, vitamins, and growth factors , colony-stimulating factors, nutrient transport molecules (transferrin), and / or cell-binding peptides , proteins, small molecules attached to albumin, polymers, dendrimers, liposomes, nanoparticles Cell-binding agents / molecules selected from the group consisting of nanoparticles, vesicles, or (viral) capsids is;

[0013] L1 and L2 are chains of atoms selected from C, N, O, S, Si, and P, and are preferably or 0 to 500 atoms, and is covalently bonded to W and V1, and V1 and V2. The atoms used to form 1 and L2 are alkylene, alkenylene, and alkynylene, Ethers, polyoxyalkylenes, esters, amines, imines, polyamines, hydrazines , hydrazones, amides, ureas, semicarbazides, carbazides, alkoxyamines, alkoxyamines Xylamine, urethane, amino acid, peptide, acyloxylamine, hydroxamic acid, or are combined in all chemically relevant ways to form these combinations. Preferably, L1 and L2 are O, NH, N, S, P, NNH, NHNH, N(R3), N(R3)N(R 3’ ), CH, CO, C(O)NH, C(O)O, NHC (O)NH, NHC(O)O, or formula (OCH2CH2) p OR3, (OCH2CH -(CH3)) p OR3, NH(CH2CH2O) p R3, NH(CH2CH(CH3) O) p R3, N[(CH2CH2O) p R3]-[(CH2CH2O) p’ R 3’ ], ( OCH2CH2) p COOR3, CH2CH2(OCH2CH2) p COOR3(p and p' is an independently selected integer from 0 to about 1000; or a combination thereof; C1-C8 alkyl; C2-C8 heteroalkyl, alkylsilyl Cycloalkyl, heterocycloalkyl; C3-C8 aryl, Ar-alkyl, heterocycle , carbocycle, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heterocyclic or natural or unnatural amino acids, or identical or different dipeptides, Tripeptides, tetrapeptides, pentapeptides, hexapeptides, heptapeptides, A ctapeptide, nonapeptide, decapeptide, undecapeptide, or dodecapeptide (Aa) consists of an array of dot units r , r=1~12 (1 to 12 amino acid units) Selected;

[0014] W is an extender unit, typically a self-immolative spacer, a peptide unit, a hydrazone, disulfide, thioether, ester, or amide bond; w is 1, 2, or 3 be;

[0015] V1 and V2 are independently O, NH, S, C1-C8 alkyl, C2-C8 heteroaryl. alkyl, alkenyl, or alkynyl, C3-C8 aryl, heterocycle, carbocycle, cycloalkane alkyl, alkylcycloalkyl, heterocycloalkyl, heteroaralkyl, heteroaralkyl alkylcycloalkyl, or alkylcarbonyl, or (Aa)r (r=1 to 12 (1 to 1 (Aa)r is a natural or unnatural amino acid, or a mixture of the same or different amino acids. are dipeptides, tripeptides, tetrapeptides, pentapeptides, and hexapeptides with different sequences. peptide, heptapeptide, octapeptide, nonapeptide, decapeptide, undecapeptide ), or (CH2CH2O) p (p= 0 to 1000); v1 and v2 are independently 0, 1 or 2, provided that v1 and v2 are simultaneously 0; if v1 or v2 is 0, means that one of the strands Q1 or Q2 fragments is absent;

[0016] Q1 and Q2 are independently represented by the formula (I-q1): [ka]

[0017] During the ceremony, [ka] is the site that binds to L1 or L2;

[0018] G1 and G2 are independently OC(O), NHC(O), C(O), CH2, NH, OC (O)NH, NHC(O)NH, O, S, B, P(O)(OH), NHP(O)(OH) , NHP(O)(OH)NH, CH2P(O)(OH)NH, OP(O)(OH)O, C H2P(O)(OH)O, NHS(O)2, NHS(O)2NH, CH2S(O)2NH , OS(O)2O, CH2S(O)2O, Ar, ArCH2, ArO, ArNH, ArS , ArNR1, or (Aa)r (r = 1 - 12);

[0019] X1 and X2 are independently O, CH2, S, NH, N(R1), + NH(R1), + N( R1)(R2), C(O), OC(O), OC(O)O, OC(O)NH, NHC(O) NH;

[0020] Y2 is O, NH, NR1, CH2, S, Ar;

[0021] G3 is OH, SH, OR1, SR1, OC(O)R1, NHC(O)R1, C(O)R 1, CH3, NH2, NR1, + NH(R1), + N(R1)(R2), C(O)OH, C(O)NH2, NHC(O)NH2, BH2, BR1R2, P(O)(OH)2, NH P(O)(OH)2, NHP(O)(NH2)2, S(O)2(OH), (CH2) q1 C(O)OH, (CH2) q1 P(O)(OH)2, C(O)(CH2) q1 C(O)O H, OC(O)(CH2) q1 C(O)OH, NHC(O)(CH2) q1 C(O)OH , CO(CH2) q1 P(O)(OH)2, NHC(O)O(CH2) q1 C(O)OH , OC(O)NH(CH2) q1 C(O)OH, NHCO(CH2) q1 P(O)(OH )2, NHC(O)(NH)(CH2) q1 C(O)OH, CONH(CH2) q1 P( O)(OH)2, NHS(O)2(CH2)q1 C(O)OH, CO(CH2) q1 S( O)2(OH), NHS(O)2NH(CH2) q1 C(O)OH, OS(O)2NH( CH2) q1 C(O)OH, NHCO(CH2) q1 S(O)2(OH), NHP(O) (OH)(NH)(CH2) q1 C(O)OH, CONH(CH2) q1 S(O)(OH ), OP(O)(OH)2, (CH2) q1 P(O)(NH)2, NHS(O)2(OH ), NHS(O)2NH2, CH2S(O)2NH2, OS(O)2OH, OS(O)2 OR1, CH2S(O)2OR1, Ar, ArR1, ArOH, ArNH2, ArSH, ArNHR1, or (Aa) q1 is;

[0022] p1, p2, and p3 are independently 0 to 100, but are not all 0 at the same time.

[0023] q1 and q2 are independently 0 to 24.

[0024] Preferably, Q1 and Q2 are independently C2 to C 90 Polycarboxylic acid or C2-C 90 Polyalkylamines, C6-C 90 Oligosaccharides or polysaccharides; C6-C 90 Zwitterionic Betaine or zwitterionic poly(sulfonates) containing quaternary ammonium cations and sulfonate anions. poly(lactic acid / glycolic acid) (PL) GA), poly(acrylate), chitosan, N-(2-hydroxypropyl)methacrylate Amide copolymer, poly[2-(methacryloyloxy)ethylphosphorylcholine]( PMPC), poly-L-glutamic acid, poly(lactide-co-glycolide) (PLG), Poly(lactide-co-glycolide), poly(ethylene glycol) (PEG), poly(propylene glycol) propylene glycol) (PPG), poly(lactide-co-glycolide), poly(ethylene glycol) modified peptides, poly(ethylene glycol) containing amino acids or peptides; Poly(ethylene glycol)-modified lipids, poly(ethylene glycol)-modified alkyl carbo Phosphoric acid, poly(ethylene glycol) modified alkylamine, poly(lactide-co-glycol) , hyaluronic acid (HA) (glycosaminoglycan), heparin / heparan sulfate (HSG) AG), chondroitin sulfate / dermatan sulfate (CSGAG), poly(ethylene glycol) poly(ethylene glycol)-modified alkyl sulfate, poly(ethylene glycol)-modified alkyl phosphate, or poly(ethylene glycol)-modified alkyl quaternary ammonium salts, etc.);

[0025] D is a tubulysin analogue having the following formula (II), or a pharmaceutically acceptable salt, water or hydrated salts; or polymorphic crystal structures; or their optical isomers, racemates, dimers, They are asteomers or enantiomers: is: [ka]

[0026] During the ceremony, [ka] is a binding site that independently binds to W;

[0027] In the formula, R 1 , R 2 , R 3, and R 4 are independently H, C1-C8 alkyl; C2-C8 Heteroalkyl or heterocyclic; C3-C8 aryl, Ar-alkyl, cycloalkyl , alkylcycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, carbon ring, or alkylcarbonyl; or R 1 R 2 , R 1 R 3 , R 2 R 3 , R 3 R 4 , R 5 R 6 , R 11 R 12 , or R 13 R 14 is a 3- to 7-membered carbocyclic ring, cycloalkyl, heterocyclic forming a ring, heterocycloalkyl, aromatic, or heteroaromatic ring system; R 1 and R 2 teeth, When independently or simultaneously binding to W, Y may not be present independently, 1 is N or CH is;

[0028] In the formula, R 5 , R 6 , R 8 , R 10 , and R 11 are independently H or C1-C4 alkyl or heteroalkyl;

[0029] In the formula, R 7 are H and R independently 14 , -R 14 C(=O)X 1 R 15 ; or -R 14 X 1 R 15 and X 1 is O, S, SS, NH, CH2, or NR 14 is;

[0030] In the formula, R 9 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 OP(=O )(OR 15 ) selected from;

[0031] In the formula, R 11 are H and R independently 14 , -R 14 C(=O)R 16 , -R 14 X 2 R 16 , -R 14 C(=O)X 2 and X 2 -O-, -S-, -NH-, -N(R 14 )- , -OR 14 -, -SR 14 -, -S(=O)-R 14 - or -NHR 14 is ;

[0032] In the formula, R 12 is R 15 , -OH, -SH, -NH2, NH, NHNH2, -NH(R 1 5 ), -OR 15 , -R 15 COR 16 , -R 15 COOR 16 , -R 15 C(O)NH 2, -R 15 C(O)NHR 17 , -SR 16 , R15 S(=O)R 16 , -R 15 P( =O)(OR 17 )2, -R 15 OP(=O)(OR 17 )2, -CH2OP(=O)( OR 17 )2, -R 15 SO2R 17 , -R 15 X 2 R 16 , -R 15 C(=O)X 2 , where X 2 are -O-, OH, SH, -S-, NH2, -NH-, -N(R 15 )-, -O -R 15 -, -SR 15 -, -S(=O)-R 15 -, CH2, or -NHR 15 -in be;

[0033] R 13 and R 14 are independently H, O, S, NH, and N(R 15 ), NHNH, -OH, - SH, -NH2, NH, NHNH2, -NH(R 15 ), -OR 15 , CO, -COX 2 , -COX 2 R 16 , R 17 , F, Cl, Br, I, SR 16 , N.R. 16 R 17 , N=N R 16 , N=R 16 , NO2, SOR 16 R 17 , SO2R 16 , SO3R 16 , OSO 3R 16 , PR 16 R 17 , POR 16 R 17 , PO2R16 R 17 , OP(O)(OR 17 )2, OCH2OP(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-(C4~C 12 glycoside), -N-( C4~C 12 Glycoside; C1-C8 alkyl, heteroalkyl; C2-C8 alkene aryl, alkynyl, heteroalkyl, heterocycloalkyl; C3-C8 aryl, A r-Alkyl, carbocycle, cycloalkyl, heteroalkylcycloalkyl, alkylcarbocycle aryl, heteroaryl, or ester, ether, or amino group having 2 to 8 carbon atoms or 1 to 8 amino acids (NH(Aa) 1~8 or CO(Aa) 1~8 (N-terminal or C-terminal or a peptide containing 1 to 8 identical or different amino acids at either end; or a peptide of the formula (OCH2CH2) p or (OCH2CH(CH3)) p polyethyleneoxy units (p is 0 to about 10 00), or a combination of the above groups; X 2 are O, S, SS, NH, C H2, OH, SH, NH2, CHR 14 , or NR 14 is;

[0034] R 15 , R 16 , and R 17 are independently H, C1-C8 alkyl, heteroalkyl C2-C8 alkenyl, alkynyl, heteroalkyl, heterocycloalkyl; C3 ~C8 aryl, Ar-alkyl, carbocycle, cycloalkyl, heteroalkylcycloa alkyl, alkylcarbonyl, heteroaryl, alkylcarbonyl, or Na + , K. + , Cs + , Li + , Ca 2+ , Mg + , Zn 2+ , N + (R1)(R2)(R3)(R4 ), HN + (C2H5OH)3 salt;

[0035] Y 1 and Y 2 are independently N or CH; q is 0 or 1; when q=0, Y 3 does not exist, and Y 4 , Y 5 , Y 6 , and Y 7 are independently CH, N, NH, O, S, or N (R 1 ), and therefore, Y 2 , Y 4 , Y 5 , Y 6 , and Y 7 are furan, pyrrole, and thio Phenol, thiazole, oxazole and imidazole, pyrazole, triazole, tetraazole When q=1, Y forms a heteroaromatic ring of thiadiazole or thiadiazole; 3 , Y 4 , Y 5 , Y 6 , and Y 7 are independently CH or N, and therefore, Y 2 , Y 3 , Y 4 , Y 5 , Y 6 , and Y 7Benzene, pyridine, pyridazine, pyrimidine, pyrazine, triazine, tetraazine It forms an aromatic ring of thorazine and pentazine.

[0036] In another embodiment of the present invention, the conjugate containing the side chain linkage is represented by formula (III): [ka]

[0037] In the formula, D, W, w, L1, L2, Q1, Q2, V1, V2, v1, v2, n, and T are the formulas This is the same definition as (I).

[0038] In another embodiment of the present invention, the side chain-linked compound is represented by formula (IV), and easily binds to the cell-binding molecule T. to form a conjugate of formula (I): [ka]

[0039] In the formula, D, W, w, L1, L2, Q1, Q2, V1, V2, v1, v2, and n are represented by the formula The definition is the same as in (I); Lv1 is a functional group described below.

[0040] In another embodiment of the present invention, the side chain-linked compound is represented by formula (V), and can be easily linked to a cell-binding molecule T. can react to form a conjugate of formula (III): [ka]

[0041] In the formula, D, W, w, L1, L2, Q1, Q2, V1, V2, v1, v2, and n are represented by the formula (I) is the same definition as

[0042] Lv1 and Lv2 are thiols, amines, carboxylic acids, selenols, and Lv1 and Lv2 are reactive groups that can react with phenol or hydroxyl groups. F, Cl, Br, I; nitrophenol; N-hydroxysuccinimide (NHS); phenol; dinitrophenol; pentafluorophenol; tetrafluorophenol ;Difluorophenol;Monofluorophenol;Pentachlorophenol;Trifluorophenol Imidazole; Dichlorophenol; Tetrachlorophenol; 1-Hydroxybenzyl Tosylate; Mesylate; 2-Ethyl-5-phenylisoxazolidinium methyl-3'-sulfonate, self-formed or other anhydrides, e.g. acetic anhydride or or anhydrides formed with formic anhydride; or for peptide coupling reactions or an intermediate molecule generated by a condensation reagent for the Mitsunobu reaction. The condensation reagent was EDC (N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide). DCC (dicyclohexylcarbodiimide), N,N'-diisopropylcarbodiimide Diimide (DIC), N-cyclohexyl-N'-(2-morpholinoethyl)carbodiimide Midomeso-p-toluenesulfonate (CMC, or CME-CDI), 1,1'-calcium Bornyldiimidazole (CDI), TBTU (O-(benzotriazol-1-yl)- N,N,N',N'-tetramethyluronium tetrafluoroborate), N,N,N' ,N'-Tetramethyl-O-(1H-benzotriazol-1-yl)uronium hexa Fluorophosphate (HBTU), (benzotriazol-1-yloxy)tris( Dimethylamino)phosphonium hexafluorophosphate (BOP), (benzotriazole (1-isopropyl-1-pyrrolidino)phosphonium hexafluorophosphate (P yBOP), diethyl cyanophosphonate (DEPC), chloro-N,N,N',N'- Tetramethylformamidinium hexafluorophosphate, 1-[bis(dimethylamino) [amino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-ol oxide hexafluorophosphate (HATU), 1-[(dimethylamino)(morpholino) methylene]-1H-[1,2,3]triazolo[4,5-b]pyridin-1-ium -3-Oxidohexafluorophosphate (HDMA), 2-chloro-1,3-dimethyl Imidazolidinium hexafluorophosphate (CIP), chlorotripyrrolidinium phosphate Sulfonium hexafluorophosphate (PyCloP), fluoro-N,N,N',N '-Bis(tetramethylene)formamidinium hexafluorophosphate (BTFF H), N,N,N',N'-tetramethyl-S-(1-oxide-2-pyridyl)thiolo Nitrium hexafluorophosphate, O-(2-oxo-1(2H)pyridyl)-N,N ,N',N'-Tetramethylthiuronium tetrafluoroborate (TPTU), S-( 1-Oxido-2-pyridyl)-N,N,N',N'-tetramethylthiuronium tetrachloride Fluoroborate, O-[(ethoxycarbonyl)cyanomethyleneamino]-N,N,N ',N'-Tetramethyluronium hexafluorophosphate (HOTU), (1-chloroisothiazolinone) Ano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholino -Carbenium hexafluorophosphate (COMU), O-(benzotriazole- 1-yl)-N,N,N',N'-bis(tetramethylene)uronium hexafluorophosphate sulfate (HBPyU), N-benzyl-N'-cyclohexylcarbodiimide (polymerization Dipyrrolidino(N-succinimidyloxy)-carbamate nium hexafluorophosphate (HSPyU), chlorodipyrrolidinocarbenium PyCIU, 2-chloro-1,3-dimethylimidazolin C1B, (benzotriazol-1-yloxy)dipyridine Peridinocarbenium hexafluorophosphate (HBPipU), O-(6-chloro Benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrachloride Fluoroborate (TCTU), Bromotris(dimethylamino)phosphonium hexafluorophosphate Bromophosphate (BroP), Propylphosphonic Anhydride (PPACA), T3P ( (Registered trademark), 2-morpholinoethyl isocyanide (MEI), N,N,N',N'-tetramethylisocyanide Tetramethyl-O-(N-succinimidyl)uronium hexafluorophosphate (HS TU), 2-bromo-1-ethyl-pyridinium tetrafluoroborate (BEP), O -[(ethoxycarbonyl)cyanomethyleneamino]-N,N,N',N'-tetramethyl Fluorocarbonyl tetrafluoroborate (TOTU), 4-(4,6-dimethoxy-1,3 ,5-triazin-2-yl)-4-methylmorpholinium chloride (MMTM,DMT MM), 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 tet Trifluoroborate (TDBTU), 1,1'-(azodicarbonyl)dipiperidine (A DD), di-(4-chlorobenzyl) azodicarboxylate (DCAD), di-tert-butyl t-Butyl azodicarboxylate (DBAD), diisopropyl azodicarboxylate Selected from diethyl azodicarboxylate (DIAD), diethyl azodicarboxylate (DEAD). may be an anhydride formed by the acid itself or by other C1-C8 anhydrides.

[0043] The present invention further provides a method for preparing the cell-binding molecule-drug conjugates of formula (I) and formula (III). and the application of the conjugates of formula (I) and formula (III). [Brief explanation of the drawings]

[0044] [Figure 1] A general synthesis of the Tuv component of a tubulysin analog is shown. [Figure 2] 1 shows the synthesis of tubulysin components. [Figure 3] 1 shows the synthesis of tubulysin components. [Figure 4] 1 shows the synthesis of tubulysin analog components. [Figure 5] 1 shows the synthesis of tubulysin analog components. [Figure 6] 1 shows the synthesis of tubulysin analog components. [Figure 7] 1 shows the synthesis of tubulysin analog components. [Figure 8] 1 shows the synthesis of tubulysin analog components containing conjugated linkers. [Figure 9] The synthesis of tubulysin analog components and their conjugation to antibodies are shown. [Figure 10] The synthesis of the side chain linker components is shown. [Figure 11] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 12] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 13]1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 14] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 15] The synthesis of the side chain linker component and its attachment to the Tup component are shown. [Figure 16] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 17] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 18] 1 shows the synthesis of tubulysin analog components. [Figure 19] 1 shows the synthesis of tubulysin analog components. [Figure 20] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 21] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 22] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 23] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 24] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 25] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 26] 1 shows the synthesis of tubulysin components containing side chain linkers. [Figure 27] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies via thiol pairs in the antibody. [Figure 28] 1 shows the synthesis of tubulysin components containing side chain linkers. [Figure 29] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies via thiol pairs in the antibody. [Figure 30]1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies via thiol pairs in the antibody. [Figure 31] 1 shows the synthesis of tubulysin components. [Figure 32] 1 shows the synthesis of tubulysin components. [Figure 33] 1 shows the synthesis of tubulysin components. [Figure 34] 1 shows the synthesis of tubulysin components containing side chain linkers. [Figure 35] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 36] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 37] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 38] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 39] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 40] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 41] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 42] 1 shows the synthesis of tubulysin analogues. [Figure 43] 1 shows the synthesis of tubulysin analogues. [Figure 44] 1 shows the synthesis of tubulysin analogues. [Figure 45] 1 shows the synthesis of tubulysin analogs and their conjugation to antibodies. [Figure 46] 1 shows the synthesis of tubulysin analogs and their conjugation to antibodies. [Figure 47] 1 shows the synthesis of tubulysin analogues. [Figure 48] 1 shows the synthesis of tubulysin analogs and their conjugation to antibodies. [Figure 49]1 shows the synthesis of tubulysin analogs and their conjugation to antibodies. [Figure 50] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 51] 1 shows the synthesis of tubulysin analogs and their conjugation to antibodies. [Figure 52] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 53] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their attachment to antibodies via a pair of thiols in the antibody. [Figure 54] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 55] 1 shows the synthesis of tubulysin analogs containing side chain linkers to antibodies. [Figure 56] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 57] 1 shows the synthesis of tubulysin analogues. [Figure 58] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their attachment to antibodies via a pair of thiols in the antibody. [Figure 59] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their attachment to antibodies via a pair of thiols in the antibody. [Figure 60] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their attachment to antibodies via a pair of thiols in the antibody. [Figure 61] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 62] 1 shows the synthesis of tubulysin analogs containing side chain linkers and their conjugation to antibodies. [Figure 63] Using a human gastric tumor N87 cell model, the antitumor effects of conjugate compounds 474, 486, 493, 601, 626, 637, 641, 669, 673, 680, and 692 at a single intravenous dose of 6 mg / kg are compared with those of T-DM1. [Figure 64]1 shows stability studies of conjugates with side chain linkages (680 and 692) compared to T-DM1 and a regular conjugate with a single linkage (compound 133) in mouse serum. DETAILED DESCRIPTION OF THE INVENTION

[0045] definition

[0046] "Alkyl" means an alkyl group formed by removing one or two hydrogen atoms from a carbon atom. It refers to an aliphatic hydrocarbon group or monovalent group derived from a carboxylic acid. The term "branched" means that the linear alkyl group has one or more carbon atoms. Multiple low carbon alkyl groups, such as methyl, ethyl, or propyl groups, may be attached. Specific examples of alkyl groups include methyl, ethyl, n-propyl, and isopropyl. , n-butyl, t-butyl, n-pentyl, 3-pentyl, octyl, nonyl, decyl, Cyclopentyl, cyclohexyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, 3,3-dimethylpentyl, 2 ,3,4-trimethylpentyl, 3-methylhexyl, 2,2-dimethylhexyl, 2, 4-dimethylhexyl, 2,5-dimethylhexyl, 3,5-dimethylhexyl, 2,4 -Dimethylpentyl, 2-methylheptyl, 3-methylheptyl, n-heptyl, isohexyl C1-C8 alkyl groups include unsubstituted butyl, n-octyl, and isooctyl. and may be substituted with one or more substituents (but not limited to the following substituents). The substituents include -C1 to C8 alkyl, -O-(C1 to C8 alkyl), aryl, , -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH2, -C(O )NHR', -C(O)N(R')2, -NHC(O)R', -SR', -S(O)2R' , -S(O)R', -OH, -halogen, -N3, -NH2, -NH(R'), -N(R R')2 and -CN, and R' are each independently C1 to C8 alkyl and aryl. is selected from the rules.

[0047] "Halogen" refers to a fluorine, chlorine, bromine, or iodine atom, and includes fluorine and chlorine atoms. is preferred.

[0048] "Heteroalkyl" means a group of 1 to 4 carbon atoms independently selected from the group consisting of O, S, and N. It refers to a C2-C8 alkyl substituted at a heteroatom selected from the group consisting of:

[0049] "Carbocycle" means a monocyclic ring system having 3 to 8 carbon atoms or a monocyclic ring system having 7 to 13 carbon atoms. refers to a bicyclic saturated or unsaturated ring. Monocyclic carbocyclic rings have 3 to 6 rings, more typically 5 or 6 Bicyclic carbocycles have 7 to 12 ring atoms and are bicyclic [4,5], [ Arranged as [5,5], [5,6], or [6,6], or 9-10 ring atoms and arranged as a bicyclic ring system [5,6] or [6,6]. Rings (C3~C8 carbocycles) include -cyclopropyl, -cyclobutyl , -cyclopentyl, -cyclopentadienyl, -cyclohexyl, -cyclohexenyl , -1,3-cyclohexadienyl, -1,4-cyclohexadienyl, -cycloheptyl -1,3-cycloheptadienyl, -1,3,5-cycloheptatrienyl, -cycloheptatrienyl Includes, but is not limited to, -cyclooctyl, and -cyclooctadienyl.

[0050] C3-C8 carbocycles can have 3, 4, 5, 6, 7, or refers to a saturated or unsaturated non-aromatic hydrocarbon carbocyclic compound having 8 carbon atoms. The 3-C8 carbocyclic ring may be unsubstituted or substituted with one or more substituents. The substituents include, but are not limited to, -C1 to C8 alkyl, -O-( C1-C8 alkyl), -aryl, -C(O)R', -OC(O)R', -C(O)O R', -C(O)NH2, -C(O)NHR', -C(O)N(R')2, -NHC(O) R', -SR', -S(O)R', -S(O)2R', -OH, -halogen, -N3, - -NH2, -NH(R'), -N(R')2, and -CN, where R' is each are independently selected from C1 to C8 alkyl and aryl.

[0051] "Alkenyl" refers to a straight-chain alkyl group having 2 to 8 carbon atoms in the chain and containing a carbon-carbon double bond. Alkenyl groups include, for example, ethenyl, Propenyl, n-butenyl, i-butenyl, 3-methylbut-2-enyl, n-pentenyl These include hexyl, hexylenyl, heptenyl, and octenyl.

[0052] "Alkynyl" refers to a straight-chain alkyl group having 2 to 8 carbon atoms in the chain and containing a carbon-carbon triple bond. The alkynyl group refers to an aliphatic hydrocarbon group which may be cyclic or branched. Examples of the alkynyl group include ethynyl, Propynyl, n-butynyl, 2-butynyl, 3-methylbutynyl, 5-pentynyl, n- Includes pentynyl, hexynyl, heptynyl, and octynyl.

[0053] An "alkylene" is a group formed by attaching two hydrogen atoms to the same or two different carbon atoms of a parent alkane. a saturated, C1-18, monovalent radical center derived by removing It refers to a straight or branched chain or cyclic hydrocarbon group. Typical alkylene groups include methylene ( -CH2-), 1,2-ethyl (-CH2CH2-), 1,3-propyl (-CH2CH 2CH2-), 1,4-butyl (-CH2CH2CH2CH2-), etc. Not limited to:

[0054] "Alkenylene" refers to a group that has two hydrogen atoms attached to the same or two different carbon atoms of a parent alkene. Unsaturated carbon atoms having 2 to 18 carbon atoms and two monovalent radical centers derived by removing a carbon atom Typical alkenylene groups include 1, Including, but not limited to, 2-ethylene (-CH=CH-).

[0055] An "alkynylene" is an alkylene group that is formed by attaching two hydrogen atoms to the same or two different carbon atoms of a parent alkyne. Unsaturated carbon atoms having 2 to 18 carbon atoms and two monovalent radical centers derived by removing a carbon atom Typical alkynylene groups include acetonitrile, ... Examples include, but are not limited to, ethylene, propargyl, and 4-pentynyl.

[0056] "Aryl" or Ar refers to an alkyl group having 3 to 14 carbon atoms, preferably 6 to 10 carbon atoms. "Heteroaromatic group" refers to an aromatic or heteroaromatic group consisting of one or several rings, including The term refers to a group in which one or several carbons, preferably 1, 2, 3, or 4 carbon atoms on the aromatic group are It refers to those replaced by O, N, Si, Se, P, or S, preferably O, S, and N. The term aryl or Ar also refers to the group in which one or more H atoms are independently replaced by -R', -halogen. , -OR', or -SR', -NR'R'', -N=NR', -N=R', -NR'R' ', -NO2, -S(O)R', -S(O)2R', -S(O)2OR', -OS(O) 2OR', -PR'R'', -P(O)R'R'', -P(OR')(OR''), -P Replaced by (O)(OR')(OR'') or -OP(O)(OR')(OR'') The R' and R'' are independently H, alkyl, alkenyl, alkylene, alkynyl ... is aryl, heteroalkyl, aryl, arylalkyl, carbonyl, or a pharmaceutical salt thereof. .

[0057] A "heterocycle" is a ring in which 1 to 4 ring carbon atoms are independently selected from O, N, It refers to ring systems substituted with heteroatoms from the group of S, Se, B, Si, and P. The most common heteroatoms are O, N, and S. Heterocycles are described in The Handbook of Chemistry and Ph , 78th Edition, CRC Press, Inc., 1997-1998, pp. 225-226, and the disclosure thereof are incorporated herein by reference. Preferred non-aromatic heterocycles include, but are not limited to: However, epoxy, aziridinyl, thiranyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl Lysinyl, oxiranyl, tetrahydrofuranyl, dioxolanyl, tetrahydropyranyl dioxanyl, dioxolanyl, piperidyl, piperazinyl, morpholinyl, pyranilinyl Imidazolinyl, pyrrolinyl, pyrazolinyl, thiazolidinyl, tetrahydropyranyl dihydropyranyl, tetrahydropyridyl, dihydropyridyl, tetrahydropyriminyl dihydrothiopyranyl, dihydrothiopyranyl, azepanyl, and condensation resulting from condensation with a phenyl group The system is included.

[0058] The term "heteroaryl" or aromatic heterocycle refers to a 5- to 14-membered, preferably 5- to 10-membered, aromatic heterocycle. Aromatic heterocyclic, monocyclic, bicyclic, or polycyclic rings, examples of which include pyrrolyl, pyridyl, and the like. , pyrazolyl, thienyl, pyrimidinyl, pyrazinyl, tetrazolyl, indolyl, quino Linyl, purinyl, imidazolyl, thienyl, thiazolyl, benzothiazolyl, furanyl , benzofuranyl, 1,2,4-thiadiazolyl, isothiazolyl, triazolyl, tetrazolyl Isoquinolyl, benzothienyl, isobenzofuryl, pyrazolyl, carbazolyl benzimidazolyl, isoxazolyl, pyridyl-N-oxide, and phenyl groups This includes condensed systems resulting from the condensation of

[0059] "Alkyl," "cycloalkyl," "alkenyl," "alkynyl," "aryl" "Heteroaryl", "heterocyclic" etc. have two hydrogen atoms. The corresponding "alkylene," "cycloalkylene," and "cycloalkylene" groups are formed by removing atoms. ", "alkenylene", "alkynylene", "arylene", "heteroarylene", " It also refers to heterocyclic groups, etc.

[0060] "Arylalkyl" refers to an alkyl group consisting of a carbon atom, typically a terminal or sp 3 Water bonded to a carbon atom It refers to an acyclic alkyl group in which one of the carbon atoms is substituted with an aryl group. The alkyl groups include, but are not limited to, benzyl, 2-phenylethan-1-yl , 2-phenylethen-1-yl, naphthylmethyl, 2-naphthylethan-1-yl, 2 -naphthylethen-1-yl, naphthobenzyl, 2-naphthophenylethan-1-yl, etc. Includes:

[0061] "Heteroarylalkyl" refers to a heteroaryl group consisting of a carbon atom, typically a terminal or sp 3 Attached to a carbon atom It refers to an acyclic alkyl group in which one of the hydrogen atoms is replaced by a heteroaryl group. Typical heteroarylalkyl groups include, but are not limited to, 2-benzimidazoline. These include 2-furylmethyl, 2-furylethyl, and the like.

[0062] Examples of "hydroxy protecting groups" include, but are not limited to, methoxymethyl ether, 2-Methoxyethoxymethyl ether, tetrahydropyranyl ether, benzyl ether p-Methoxybenzyl ether, Trimethylsilyl ether, Triethylsilyl ether ter, triisopropylsilyl ether, t-butyldimethylsilyl ether, triphenyl Nylmethylsilyl ether, acetate ester, substituted acetate ester, pivaloate, benzoate These include methylsulfonate, methanesulfonate, and p-toluenesulfonate.

[0063] A "leaving group" refers to a functional group that can be displaced by another functional group. Suitable leaving groups are well known in the art and include, but are not limited to, halo genides (e.g., chlorides, bromides, and iodides), methanesulfonyl (mesyl), p -toluenesulfonyl (tosyl), trifluoromethylsulfonyl (triflate), and and trifluoromethylsulfonate. Preferred leaving groups include nitrophenone and N-hydroxysuccinimide (NHS); phenol; dinitrophenol; phenol Pentafluorophenol; Tetrafluorophenol; Difluorophenol; Monofluoro Pentachlorophenol; Triflate; Imidazole; Dichlorophenol phenol;tetrachlorophenol;1-hydroxybenzotriazole;tosylate;methyl silylates; 2-ethyl-5-phenylisoxazolium-3'-sulfonate, self- or or acid anhydrides formed with other acid anhydrides (e.g., acetic anhydride, formic anhydride); or peptides The intermediates produced by condensation reagents for the amide coupling reaction or the Mitsunobu reaction is selected from the interbody.

[0064] The following abbreviations may be used herein and have the definitions indicated below: Boc, tert-Butoxycarbonyl; BroP, Bromotrispyrrolizinophosphonium hexa Fluorophosphate; CDI, 1,1'-carbonyldiimidazole; DCC, dicyclohexyl Dihexylcarbodiimide; DCE, 1,2-dichloroethane; DCM, dichloromethane ;DIAD, diisopropyl azodicarboxylate;DIBAL-H, hydrogenated diisobutyl aldehyde Aluminum; DIPEA, diisopropylethylamine; DEPC, diethylphosphoric acid didiate; DMA, N,N-dimethylacetamide; DMAP, 4-(N,N-dimethyl (amino)pyridine; DMF, N,N-dimethylformamide; DMSO, dimethyl sulfoxide dithioester; DTT, dithioester; EDC, 1-(3-dimethylaminopropyl)-3 -ethylcarbodiimide hydrochloride; ESI-MS, electrospray mass spectrometry; HATU , O-(7-azabenzotriazol-1-yl)-N,N,N'-N'-tetramethyl Uronium hexafluorophosphate; HOBt, 1-hydroxybenzotriazole; H PLC, high-pressure liquid chromatography; NHS, N-hydroxysuccinimide; MMP , 4-methylmorpholine; PAB, p-aminobenzyl; PBS, phosphate-buffered saline (pH 7.0-7.5); PEG, polyethylene glycol; SEC, size exclusion chromatography TCEP, tris(2-carboxyethyl)phosphine; TFA, triflate Fluoroacetic acid; THF, tetrahydrofuran; Val, valine.

[0065] "Amino acids" can be natural and / or unnatural amino acids, preferably α-amino acids Natural amino acids are those encoded by the genetic code and include alanine, arginine, Asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, Styridine, isoleucine, leucine, lysine, methionine, phenylalanine, proline , serine, threonine, tyrosine, tryptophan, and valine. are derived forms of protein-forming amino acids, e.g., hydroxyproline, lanthionine 2-aminoisobutyric acid, dehydroalanine, γ-aminobutyric acid (neurotransmitter), ornithin Citrulline, β-alanine (3-aminopropanoic acid), γ-carboxyglutamate Selenocysteine ​​(present as in most eukaryotes, but not directly linked to DNA) (not present in some archaea and one bacterium), pyrrolysine (only present in some archaea and one bacterium), N-phosphatase methylmethionine (often the first amino acid in proteins in bacteria, mitochondria, and chloroplasts) amino acid), 5-hydroxytryptophan, L-dihydroxyphenylalanine, triiodothyronine Dothyronine, L-3,4-dihydroxyphenylalanine (DOPA), and O-phosphoric acid The term amino acid also includes amino acid analogs and mimetics. is the same as that of a natural amino acid, except that the R group is not one found in a natural amino acid. These are compounds with the same general structure: H2N(R)CHCO2H. Examples of analogs include Moselin, norleucine, methionine sulfoxide, and methionine methyl sulfonium Preferably, the amino acid mimetic has a structure different from the general chemical structure of an α-amino acid. The term "unnatural amino acid" refers to a compound that has the structure, but functions similarly to, It is intended to represent the "D" stereochemical form, whereas naturally occurring amino acids are in the "L" form. When 1 to 8 amino acids are used in the present application, the amino acid sequence is selected from the cleavage recognition sequences of proteases. Many cleavage recognition sequences are known in the art, for example, Matayoshi et al. Science 247: 954 (1990); Dunn et al. Meth. Enzymol. 241: 25 4 (1994);Seidah et al. Meth. Enzymol. 244: 175 (1994);Thornberry, Meth. Enzymo l. 244: 615 (1994);Weber et al. Meth. Enzymol. 244: 595 (1994);Smith et al. Me th. Enzymol. 244: 412 (1994); and Bouvier et al. Meth. Enzymol. 248: 614 (1995) See, the disclosure of which is incorporated herein by reference. In particular, the sequence is Val-Cit, Ala-Val, Ala-Ala, Val-Val, Val-Ala-Val, Lys- Lys, Ala-Asn-Val, Val-Leu-Lys, Cit-Cit, Val- From the group consisting of Lys, Ala-Ala-Asn, Lys, Cit, Ser, and Glu be selected.

[0066] A "glycoside" is a compound in which a sugar group is bonded to another group via its anomeric carbon through a glycosidic bond. Glycosides are molecules that are linked together. Glycosides are O-(O-glycosides), N-(glycosylamines), ), S-(thioglycoside), or C-(C-glycoside) glycosidic bonds The core empirical formula is C m (H2O) n (where m is different from n, m and n are <36), where glycosides include glucose (dextrose ), fructose (levulose), allose, altrose, mannose, gulose, yogurt Dose, galactose, talose, galactosamine, glucosamine, sialic acid, N-acetylglucosamine Glucosamine, sulfoquinovose (6-deoxy-6-sulfo-D-glucopyranose) sucrose), ribose, arabinose, xylose, lyxose, sorbitol, mannitol , sucrose, lactose, maltose, trehalose, maltodextrin, raffinose It contains stachyose, glucuronic acid (glucuronide), and stachyose. L-form, 5-atom furanose-form, 6-atom cyclic pyranose-form, or acyclic, α-isomer (-OH of the anomeric carbon below the plane of the carbon atoms in the Haworth projection), or β-isomer (Haworth It can be the -OH of the anomeric carbon above the plane of the Worth projection. The term "ol" is used herein to refer to an oligosaccharide containing 3 to 6 sugar units.

[0067] As used herein, the term "antibody" refers to a full-length immunoglobulin molecule or a full-length immunoglobulin molecule. The immunologically active portion of an immunoglobulin molecule, i.e., the portion of the immunoglobulin molecule that is immunized against an important target antigen or part thereof. Antigen-binding sites that specifically bind to a target include, but are not limited to: However, it does not include cancer cells or cell populations that produce autoimmune antibodies associated with autoimmune diseases. The immunoglobulins disclosed herein include any immunoglobulin molecule. any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or Immunoglobulins can be of any subclass. Preferably, the immunoglobulin is of human, murine, or rabbit origin. Antibodies useful in the present invention are preferably monoclonal, and may be polyclonal, monoclonal, or both. monoclonal, bispecific, human, humanized or chimeric antibodies, single chain antibodies, Fv, Fab fragments , F(ab') fragments, F(ab')2 fragments, and Fab expression libraries Antibody fragments, anti-idiotypic (anti-Id) antibodies, CDRs, and cancer cell Any of the above epitopes that immunospecifically bind to a cytoplasmic antigen, a viral antigen, or a microbial antigen. These include, but are not limited to, loop-binding fragments.

[0068] The left and right hands are the same except that they are opposite along one axis (just change direction). (These are also known as "optical isomers"), so the two hands do not appear identical. "Enantiomers" are two substances that are non-superimposable (not identical) mirror images of each other. A single chiral atom or similar structural feature in a compound allows The compound has two non-superimposable structures, each a mirror image of the other. The presence of multiple chiral elements in a particular compound increases the number of possible geometric configurations. A pure enantiomer is one that is, within the limits of detection, When present in a symmetric environment, enantiomers are Identical chemical and physical properties except for their ability to rotate plane polarized light (+ / -) by equal amounts and in opposite directions. (However, polarized light can be considered as an asymmetric medium.) A mixture of equal parts of an optically active isomer and its enantiomer is called a It is called semicircular and has zero net rotation of plane polarized light. This means that the positive rotation of each (+) sign is This is because the negative rotation of the (-) form exactly cancels out. often undergo different chemical reactions than other enantiomeric substances. Because they are enantiomers, there can be significant differences in the effects of the two enantiomers on living organisms. For example, in drugs, often only one of the drug's enantiomers produces the desired physiological effect. the other enantiomer is less active, inactive, or possibly has adverse effects This discovery led to the discovery of a compound that consists of only one enantiomer ("enantiopure"). By developing drugs that can enhance pharmacological effects and sometimes eliminate some side effects, .

[0069] Isotopes are variations of a particular chemical element that differ in the number of neutrons. All isotopes of a particular element has the same number of protons in each atom. Each atomic number identifies a specific element, but isotopes No distinction. Atoms of a given element may have a wide range of neutron numbers. The number of atoms (both protons and neutrons) is the atom's mass number, and each isotope of a particular element has a different mass. For example, carbon-12, carbon-13, and carbon-14 have mass numbers 12 and 14, respectively. The atomic number of carbon is 6. This means that all carbon atoms are Since the atom has six protons, the neutron counts of these isotopes are 6, 7, and 8. The hydrogen atoms include protium ( 1 H), deuterium ( 2 H), and tritium ( 3 H ) isotopes, deuterium has twice the mass of protium, and tritium has three times the mass of protium. The mass of uranium is 1. By using isotopic substitution, we can determine the mechanism of chemical reactions. Isotope substitution can occur via substitution of substances in the body. metabolic changes (e.g., substitution of cytochrome P450 or glucuronosyltransferase enzymes) The mechanism of absorption and distribution of certain xenobiotics is not only via metabolic enzymes but also via the mechanisms of absorption and distribution after administration. / Investigate how the body reacts to chemicals and the excretion pathways and effects of drug metabolites This study is called pharmacokinetics (PK). It can be used to study the biochemical and physiological effects of animals (including humans), microorganisms, and This study includes those occurring in combination with other organisms (e.g., infections). The effects can be caused by animals (including humans), microorganisms, or combinations of organisms (infectious diseases). These include those that appear in the cerebrospinal fluid (e.g., steroids, steroid hormones, etc.), which together affect the administration, benefits, and adverse effects of a drug. Isotopes include stable (non-radioactive) or unstable elements. Isotopic substitution of drugs may have a different therapeutic effect than the original drug.

[0070] "Pharmaceutically" or "pharmaceutically acceptable" means that the corresponding compound or compound composition is suitable for are harmful, allergic or produce other adverse reactions when administered to animals or humans in an appropriate manner; It refers to the inability to do so.

[0071] A "pharmaceutically acceptable solvate" or "solvate" is disclosed as a compound containing one or more solvent molecules. Examples of solvents that form pharmacologically acceptable solvates include water. , isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanol These include, but are not limited to, phenolamines.

[0072] Pharmaceutically acceptable auxiliary materials include any carrier, diluent, adjuvant or excipient, e.g. , preservatives, antioxidants, fillers, disintegrants, wetting agents, emulsifiers, suspending agents, solvents, dispersion media, These include antibacterial agents, antifungal agents, isotonic agents, and absorption delaying agents. It is common practice to add these auxiliary materials to drug components that have the desired properties. It is reasonable to add auxiliary materials to the active pharmaceutical ingredient, unless the auxiliary materials are incompatible with the active pharmaceutical ingredient. It can be said that in order to achieve good results, active auxiliary materials are added to the drug components. Good too.

[0073] In the present invention, the term "pharmaceutical salt" refers to a salt derivative of the compound of the present invention. By suitable modification, the compounds of the present invention can be formed into the corresponding acid or alkali salts. Possible salts include the conventional non-toxic salts or quaternary ammonium salts, which are not intended to be limiting unless otherwise specified. The compound can be prepared by using a non-toxic inorganic or organic acid corresponding to the compound. Examples of the acid include hydrochloric acid, hydrobromic acid, sulfuric acid, aminosulfonic acid, phosphoric acid, and nitric acid, and the organic acid Acids include acetic acid, propionic acid, succinic acid, tartaric acid, citric acid, methanesulfonic acid, benzoic acid, Toluene sulfonic acid, glucuronic acid, glutamic acid, benzoic acid, salicylic acid, toluene sulfonate Acids that are used in pharmaceutically acceptable salts include carboxylic acids, oxalic acids, fumaric acids, and lactic acids. Other salts include trometamol, meglumine, and pyrroleethanol. ammonium salts such as ammonium salts, and sodium, potassium, calcium, zinc, magnesium, etc. Contains metal salts of

[0074] In the present invention, pharmaceutical salts are prepared by conventional chemical methods containing acidic or basic residues. Generally, these salts can be prepared from the parent compound by dissolving the salt in water, an organic solvent, or both. In a mixed solvent of the free acid or free base form of these compounds and a stoichiometric amount of an appropriate salt It can be obtained by reaction with a base or an acid. Ethers are generally used as non-aqueous reaction solvents. ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. For a list of salts, see Remington's Pharmaceutical Sciences iences”, 17th edition. Mack Publishing Company, Eas ton, PA, 1985, p. 1418, the disclosure of which is incorporated by reference. .

[0075] "Administering" or "Administration" ion) means any manner of transferring, delivering, introducing or transporting medicines or other drugs to a subject. Such modes include oral administration, topical contact, intravenous, intraperitoneal, intramuscular, lesional, nasal, and the like. Also contemplated by the present invention are intrathecal, intravenous, or intracavitary administration. Such devices may utilize active or passive transport. The delivery device may be a slow release or a fast release delivery device.

[0076] In the context of cancer, the term "treating" includes any or all of the following: Inhibition of tumor cell or cancer cell proliferation, prevention of tumor cell or cancer cell replication, reduction of tumor burden Relief and improvement of one or more symptoms associated with the disease.

[0077] In the context of autoimmune disease, the term "treat" refers to any or all of the following: Cells associated with the pathology of autoimmune diseases, including, but not limited to, cells involved in the production of autoimmune antibodies Preventing replication of potential cells, reducing the burden of autoimmune antibodies, and one or more symptoms of an autoimmune disease Improvements.

[0078] In the context of infectious diseases, the term "treat" includes the prevention of the development of the pathogen that causes the infection. prevention of the growth, proliferation, or replication of the virus, and / or amelioration of one or more symptoms of the infection. can be.

[0079] Examples of "mammals" or "animals" include humans, rats, mice, guinea pigs, monkeys, and pigs. , goats, cows, horses, dogs, cats, birds, and poultry.

[0080] The novel conjugates disclosed herein use bridge linkers. Examples of conjugates and their synthesis are shown in Figures 1 to 34.

[0081] Cell-binding agent-cytotoxic agent conjugates via side chain linkages

[0082] In one embodiment of the invention, the conjugate containing the side chain linkage is represented by formula (I): [ka]

[0083] During the ceremony, [ka] represents a single bond; n is 1 to 30;

[0084] T is an antibody, a single-chain antibody, an antibody fragment that binds to a target cell, a monoclonal antibody, a single-chain monoclonal antibody Clonal antibodies, monoclonal antibody fragments that bind to target cells, chimeric antibodies, and antibodies that bind to target cells chimeric antibody fragments, domain antibodies, domain antibody fragments, and antibodies that bind to target cells Mimicking Adnectins, DARPins, lymphokines, hormones, vitamins, and growth factors , colony-stimulating factors, nutrient transport molecules (transferrin), and / or cell-binding peptides , proteins, small molecules attached to albumin, polymers, dendrimers, liposomes, nanoparticles Cell-binding agents / molecules selected from the group consisting of nanoparticles, vesicles, or (viral) capsids is;

[0085] L1 and L2 are chains of atoms selected from C, N, O, S, Si, and P, and are preferably or 0 to 500 atoms, and is covalently bonded to W and V1, and V1 and V2. The atoms used to form 1 and L2 are alkylene, alkenylene, and alkynylene, Ethers, polyoxyalkylenes, esters, amines, imines, polyamines, hydrazines , hydrazones, amides, ureas, semicarbazides, carbazides, alkoxyamines, alkoxyamines Xylamine, urethane, amino acid, peptide, acyloxylamine, hydroxamic acid, or are combined in all chemically relevant ways to form these combinations. It's fine.

[0086] Preferably, L1 and L2 are O, NH, N, S, P, NNH, NHNH, N(R3) , N(R3)N(R 3’ ), CH, CO, C(O)NH, C(O)O, NHC(O)NH , NHC(O)O, or formula (OCH2CH2) p OR3, (OCH2CH-(CH3 )) p OR3, NH(CH2CH2O) p R3, NH(CH2CH(CH3)O) p R3 , N[(CH2CH2O) p R3]-[(CH2CH2O) p’ R 3’ ], (OCH2C H2) p COOR3, CH2CH2(OCH2CH2) p COOR3 (p and p' are independent and the unit is an integer of 0 to about 1000 selected from the group consisting of polyethyleneoxy units, Combinations; C1-C8 alkyl; C2-C8 heteroalkyl, alkylcycloalkyl Aryl, heterocycloalkyl; C3-C8 aryl, Ar-alkyl, heterocycle, carbocycle, Cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl or natural or unnatural amino acids, or the same or different dipeptides, tripeptides peptides, tetrapeptides, pentapeptides, hexapeptides, heptapeptides, octapeptides a sequence of dodecapeptide, nonapeptide, decapeptide, undecapeptide, or dodecapeptide units Consists of columns (Aa) r , r=1 to 12 (1 to 12 amino acid units) ;

[0087] W is C1~C 18 The extension unit is usually a self-immolative spacer, a peptide unit , hydrazone, disulfide, thioether, ester, or amide bond; w is 1 or 2 or 3;

[0088] V1 and V2 are independently O, NH, S, C1-C8 alkyl, C2-C8 heteroaryl. alkyl, alkenyl, or alkynyl, C3-C8 aryl, heterocycle, carbocycle, cycloalkane alkyl, alkylcycloalkyl, heterocycloalkyl, heteroaralkyl, heteroaralkyl alkylcycloalkyl, or alkylcarbonyl, or (Aa)r (r=1 to 12 (1 to 1 (Aa)r is a natural or unnatural amino acid, or a mixture of the same or different amino acids. are dipeptides, tripeptides, tetrapeptides, pentapeptides, and hexapeptides with different sequences. peptide, heptapeptide, octapeptide, nonapeptide, decapeptide, undecapeptide ), or (CH2CH2O) p (p= 0 to 1000); v1 and v2 are independently 0, 1 or 2, provided that v1 and v2 are simultaneously 0; if v1 or v2 is 0, means that one of the strands Q1 or Q2 fragments is absent;

[0089] Q1 and Q2 are independently represented by the formula (I-q1): [ka]

[0090] During the ceremony, [ka] is the site that binds to L1 or L2;

[0091] G1 and G2 are independently OC(O), NHC(O), C(O), CH2, NH, OC (O)NH, NHC(O)NH, O, S, B, P(O)(OH), NHP(O)(OH) , NHP(O)(OH)NH, CH2P(O)(OH)NH, OP(O)(OH)O, C H2P(O)(OH)O, NHS(O)2, NHS(O)2NH, CH2S(O)2NH , OS(O)2O, CH2S(O)2O, Ar, ArCH2, ArO, ArNH, ArS , ArNR1, or (Aa)q1;

[0092] G3 is OH, SH, OR1, SR1, OC(O)R1, NHC(O)R1, C(O)R 1, CH3, NH2, NR1, + NH(R1), + N(R1)(R2), C(O)OH, C(O)NH2, NHC(O)NH2, BH2, BR1R2, P(O)(OH)2, NH P(O)(OH)2, NHP(O)(NH2)2, S(O)2(OH), (CH2) q1 C(O)OH, (CH2) q1 P(O)(OH)2, C(O)(CH2) q1 C(O)O H, OC(O)(CH2) q1 C(O)OH, NHC(O)(CH2) q1 C(O)OH , CO(CH2) q1 P(O)(OH)2, NHC(O)O(CH2) q1 C(O)OH , OC(O)NH(CH2) q1 C(O)OH, NHCO(CH2) q1 P(O)(OH )2, NHC(O)(NH)(CH2) q1 C(O)OH, CONH(CH2) q1 P( O)(OH)2, NHS(O)2(CH2) q1 C(O)OH, CO(CH2) q1S( O)2(OH), NHS(O)2NH(CH2) q1 C(O)OH, OS(O)2NH( CH2) q1 C(O)OH, NHCO(CH2) q1 S(O)2(OH), NHP(O) (OH)(NH)(CH2) q1 C(O)OH, CONH(CH2) q1 S(O)(OH ), OP(O)(OH)2, (CH2) q1 P(O)(NH)2, NHS(O)2(OH ), NHS(O)2NH2, CH2S(O)2NH2, OS(O)2OH, OS(O)2 OR1, CH2S(O)2OR1, Ar, ArR1, ArOH, ArNH2, ArSH, ArNHR1, or (Aa) q1 where (Aa) q1 The same or different heavens a peptide containing a sequence of natural or unnatural amino acids;

[0093] X1 and X2 are independently O, CH2, S, S(O), NHNH, NH, N(R1), + NH(R1), + N(R1)(R2), C(O), OC(O), OC(O)O, OC(O )NH, NHC(O)NH;

[0094] Y2 is O, NH, NR1, CH2, S, NHNH, Ar;

[0095] p1, p2, and p3 are independently 0 to 100, but not simultaneously 0;

[0096] q1 and q2 are independently 0 to 24;

[0097] R1, R2, R3, and R 3’ are independently H, C1-C8 alkyl; C2-C8 hetero Aryl, Ar-alkyl, cycloalkyl, or heterocyclic; C3-C8 aryl, Ar-alkyl, cycloalkyl, ar alkylcycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, carbocycle, or alkylcarbonyl;

[0098] Preferably, Q1 and Q2 are independently C2 to C 100 Polycarboxylic acids; C2-C 90 Polyalkylamine; C6-C 90 Oligosaccharides or polysaccharides; C6-C 100 Zwitterion Betai Zwitterionic poly(sulfonates) containing sulfonate or quaternary ammonium cations and sulfonate anions (Phenylalanine) (PSB); (Poly(lactic / glycolic acid) (PLGA), Poly(acrylonitrile) acrylate), chitosan, N-(2-hydroxypropyl) methacrylamide copolymer, Poly[2-(methacryloyloxy)ethylphosphorylcholine] (PMPC), poly- L-glutamic acid, poly(lactide-co-glycolide) (PLG), poly(lactide-co-glycolide) -glycolide), poly(ethylene glycol) (PEG), poly(propylene glycol) ) (PPG), poly(lactide-co-glycolide), poly(ethylene glycol) modified polyethylene glycol Poly(ethylene glycol) containing amino acids or peptides, poly(ethylene glycol) Poly(ethylene glycol)-modified lipids, poly(ethylene glycol)-modified alkyl carboxylic acids, poly(ethylene glycol) modified alkylamine, poly(lactide-co-glycolide), hyaluronic acid ( HA) (glycosaminoglycan), heparin / heparan sulfate (HSGAG), chondroitin Dermatan sulfate / dermatan sulfate (CSGAG), poly(ethylene glycol)-modified alkyl sulfate, poly(ethylene glycol)-modified alkyl phosphate, or poly(ethylene glycol) C6-C consisting of alkyl quaternary ammonium salts modified with alkyl groups 100 biodegradable polymers -is;

[0099] Examples of structures of Q1 and Q2 are shown below: [ka] JPEG0007637431000015.jpg196165

[0100] In the formula, R 25 and R 25’ are independently H;HC(O), CH3C(O), CH3C( NH), C1-C 18 Alkyl, C1-C 18 Alkyl, alkyl-Y1-SO3H, C 1-C 18 Alkyl-Y1-PO3H2, C1-C 18 Alkyl-Y1-CO2H, C1 -C 18 Alkyl-Y1-N + R 1’ R 2’ R 3’ R 4’ , C1-C 18 Alkyl-Y 1-CONH2, C2-C 18 Alkylene, C2-C 18 Esters, C2-C 18 Aete Lu, C2-C 18 Amines, C2-C 18 Alkyl carboxylamides, C3-C 18 Ants C3-C 18 Cyclic alkyl, C3-C 18 Heterocyclic, 1-24 amino acids; C2-C1 8 lipids, C2-C 18 Fatty acids, or C2-C 18 selected from fatty ammonium lipids; X1 and X2 are independently NH, N(R1'), O, CH2, S, C(O), S(O), Selected from S(O2), P(O)(OH), NHNH, CH=CH, Ar, or (Aa)q1 where q1=0-24 (0-24 amino acids, q1=0 means absence);

[0101] X1, X2, X3, X4, Y1, Y2, and Y3 are independently NH, N(R1'), O , C(O), CH2, S, S(O), NHNH, C(O), OC(O), OC(O)O, OC(O)NH, NHC(O)NH, Ar or Ar or (Aa)q1, wherein and X1, X2, X3, X4, Y1, Y2, and Y3 may not independently exist;

[0102] p1, p2, and p3 are independently 0 to 100 and are not simultaneously 0;

[0103] q1, q2, and q3 are independently 0 to 24;

[0104] R 1’ , R 2’ , R 3’ , and R 4’ are independently selected from H and C1-C6 alkyl be;

[0105] Aa is a natural or unnatural amino acid;

[0106] Ar or (Aa)q1 are the same or different peptide sequences, and q1=0 is (Aa) This means that q1 does not exist.

[0107] D is a tubulysin analogue having the following formula (II) or a pharmaceutically acceptable salt thereof, or a polymorphic crystalline structure; or an optical isomer, racemic or diastereomeric form thereof. omers, or enantiomers: [ka]

[0108] During the ceremony, [ka] is a binding site that binds independently to W;

[0109] In the formula, R 1 , R 2 , R 3 , and R 4 are independently H, C1-C8 alkyl; C2-C8 Heteroalkyl or heterocyclic; C3-C8 aryl, Ar-alkyl, cycloalkyl , alkylcycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, carbon ring, or alkylcarbonyl; or R 1 R 2 , R 1 R 3 , R 2 R 3 , R 3 R 4 , R 5 R 6 , R 11 R 12 , or R 13 R 14 is a 3- to 7-membered carbocyclic ring, cycloalkyl, heterocyclic forming a ring, heterocycloalkyl, aromatic, or heteroaromatic ring system; R 1 and R 2 teeth, When independently or simultaneously binding to W, Y may not be present independently, 1 is N or CH is;

[0110] In the formula, R 5 , R 6 , R 8 , R 10 , and R 11 are independently H or C1-C4 alkyl or heteroalkyl;

[0111] In the formula, R 7 are H and R independently 14 , -R 14 C(=O)X 1 R 15 ; or -R 14 X 1 R 15 and X 1 is O, S, SS, NH, CH2, or NR 14 is;

[0112] In the formula, R 9 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 OP(=O )(OR 15 ) selected from;

[0113] In the formula, R 11 are H and R independently 14 , -R 14 C(=O)R 16 , -R 14 X 2 R 16 , -R 14 C(=O)X 2 and X 2 -O-, -S-, -NH-, -N(R 14 )- , -OR 14 -, -SR 14 -, -S(=O)-R 14 - or -NHR 14 is ;

[0114] In the formula, R 12 is R 15, -OH, -SH, -NH2, NH, NHNH2, -NH(R 1 5 ), -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, -CH2OP(=O)( OR 17 )2, -R 15 SO2R 17 , -R 15 X 2 R 16 , -R 15 C(=O)X 2 , where X 2 are -O-, OH, SH, -S-, NH2, -NH-, -N(R 15 )-, -O -R 15 -, -SR 15 -, -S(=O)-R 15 -, CH2, or -NHR 15 -in be;

[0115] R 13 and R 14 are independently H, O, S, NH, N(R 15 ), NHNH, -OH, -SH, -NH2, NH, NHNH2, -NH(R 15 ), -OR 15 , CO, -COX 2 , -COX 2 R 16 , R 17 , F, Cl, Br, I, SR 16 , N.R.16 R 17 , N= NR 16 , N=R 16 , NO2, SOR 16 R 17 , SO2R 16 , SO3R 16 , OS O3R 16 , PR 16 R 17 , POR 16 R 17 , PO2R 16 R 17 , OP(O)(O R 17 )2, OCH2OP(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-(C4~C 12 glycoside), -N- (C4~C 12 Glycoside; C1-C8 alkyl, heteroalkyl; C2-C8 alkyl alkynyl, alkynyl, heteroalkyl, heterocycloalkyl; C3-C8 aryl; Ar-alkyl, carbocycle, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl carbonyl, heteroaryl, or ester, ether, or alkyl group having 2 to 8 carbon atoms or 1 to 8 amino acids (NH(Aa) 1~8 or CO(Aa) 1~8 (N-terminal or C or a peptide containing 1 to 8 identical or different amino acids at the termini; or a peptide of the formula (OCH2CH2 ) p or (OCH2CH(CH3)) p polyethyleneoxy units (p is 0 to about 1 000), or a combination of the above groups; X 2 are O, S, SS, NH, CH2, OH, SH, NH2, CHR 14 , or NR 14 is;

[0116] R 15 , R 16 , and R 17 are independently H, C1-C8 alkyl, heteroalkyl C2-C8 alkenyl, alkynyl, heteroalkyl, heterocycloalkyl; C3 ~C8 aryl, Ar-alkyl, carbocycle, cycloalkyl, heteroalkylcycloa alkyl, alkylcarbonyl, heteroaryl, alkylcarbonyl, or Na + , K. + , Cs + , Li + , Ca 2+ , Mg + , Zn 2+ , N + (R 1 )(R 2 )(R 3 )(R 4 ), HN + (C2H5OH)3 salt;

[0117] Y 1 and Y 2 are independently N or CH; q is 0 or 1; when q=0, Y 3 does not exist, and Y 4 , Y 5 , Y 6 , and Y 7 are independently CH, N, NH, O, S, or N (R 1 ), and therefore, Y 2 , Y 4 , Y 5 , Y 6 , and Y 7 are furan, pyrrole, and thio Phenol, thiazole, oxazole and imidazole, pyrazole, triazole, tetraazole When q=1, Y forms a heteroaromatic ring of thiadiazole or thiadiazole; 3 , Y 4 , Y 5 , Y 6 , and Y 7 are independently CH or N, and therefore, Y 2 , Y 3 , Y 4 , Y 5 , Y 6 , and Y 7 Benzene, pyridine, pyridazine, pyrimidine, pyrazine, triazine, tetraazine It forms an aromatic ring of thorazine and pentazine.

[0118] An example of a tubulysin structure is shown below: [ka] JPEG0007637431000019.jpg228144JPEG0007637431000020.jpg242148JPEG000 7637431000021.jpg237143JPEG0007637431000022.jpg247144JPEG00076374310 00023.jpg243152JPEG0007637431000024.jpg245157JPEG0007637431000025.j pg243141JPEG0007637431000026.jpg248143JPEG0007637431000027.jpg211140

[0119] In the formula, R 20 is H; C1-C8 linear or branched alkyl or heteroalkyl; C2-C 8 straight or branched alkenyl, alkynyl, alkylcycloalkyl, heterocycloalkyl Aryl; C3-C8 linear or branched aryl, Ar-alkyl, heterocycle, carbocycle, cycloacycle alkyl, heteroalkyl, cycloalkyl, alkylcarbonyl, heteroaryl; Nate (-C(O)OR 17 ), carbamates (-C(O)NR 17 R 18 ); or carbon Carboxylic acid salts, esters, ethers, or amides having 1 to 8 atoms; or amino acids having 1 to 8 atoms or the formula (OCH2CH2) p or (OCH2CH(CH3)) p Polyethylene an oxy unit (wherein p is an integer from 0 to about 1000); or R 20 There is no acid The elements form ketones or are a combination thereof;

[0120] Z 3 and Z 3 are independently H, OH, NH2, O, NH, COOH, COO, C(O) , C(O), C(O)NH, C(O)NH2, R 18 , OCH2OP(O)(OR 18 ) 2. OC(O)OP(O)(OR 18 )2, OPO(OR 18 )2, NHPO(OR 18 )2, OP(O)(OR 18 )OP(O)(OR 18 )2, OC(O)R 18 , OC(O )NHR 18 , OSO2(OR 18 ), O-(C4-C 12 -glycosides), linear or branched Branched alkyl or heteroalkyl; C2-C8 straight or branched alkenyl, alkynyl, alkenyl alkylcycloalkyl, heterocycloalkyl; C3-C8 linear or branched aryl, A r-alkyl, heterocycle, carbocycle, cycloalkyl, heteroalkylcycloalkyl, ar Carbonyl, heteroaryl; carbonate (-C(O)OR17 ), carbamates (-C(O)NR 17 R 18 ) is;

[0121] R 17 and R 18 are independently H, C1-C8 linear or branched alkyl or heteroalkyl C2-C8 linear or branched alkenyl, alkynyl, alkylcycloalkyl, hexyl Heterocycloalkyl; C3-C8 aryl, Ar-alkyl, heterocycle, carbocycle, cyclo Alkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl; Carbonate (-C(O)OR 17 ), carbamates (-C(O)NR 17 R 18 ) is;

[0122] R 19 are H, OH, NH2, OSO2 (OR 18 ), XCH2OP(O)(OR 18 ) 2. XPO(OR 18 )2, XC(O)OP(O)(OR 18 )2, XC(O)R 18 , XC(O)NHR 18 , C1-C8 alkyl or carboxylate; C2-C8 alkenyl alkyl, alkynyl, alkylcycloalkyl, heterocycloalkyl; C3-C8 aryl or alkylcarbonyl; or a pharmaceutical salt; where X is O, S, NH, NHN H, or CH2;

[0123] R 7 has the same definition as above.

[0124] Furthermore, W, L1, L2, V1, and V2 are independently 6-maleimide having the structure shown below. Caproyl (MC), maleimidopropanoyl (MP), valine-citrulline (val- cit or vc), alanine-phenylalanine (ala-phe or af), p-amino benzyloxycarbonyl (PAB), 4-thiopentanoate (SPP), 4-(N -maleimidomethyl)cyclohexane-1-carboxylic acid ester (MCC), (4-acetonitrile 4-thio-2-methyl-aminobenzoic acid (SIAB), 4-thio-butyrate (SPDB), -hydroxysulfonyl-butyrate (2-sulfo-SPDB), or 1-12 One or more linker components of a natural or unnatural peptide having natural or unnatural amino acid units Natural amino acids may include aspartic acid, glutamic acid, arginine, hyaluronic acid, and the like. Styrene, lysine, serine, threonine, asparagine, glutamine, cysteine, selenium Cysteine, tyrosine, phenylalanine, glycine, proline, tryptophan, Preferably selected from lanine;

[0125] [ka] JPEG0007637431000029.jpg247156, and L- or D-, natural or non-natural peptides containing 1 to 20 amino acids;

[0126] W, L1, L2, V1, and V2 are independently a self-immolative or non-self-immolative component, a peptide unit, hydrazone bond, disulfide, ester, oxime, amide, or thioether The self-immolative unit may also include, but is not limited to, 2-amino Neumidazole-5-methanol derivatives, heterocyclic PAB analogues, β-glucuronides, and and o- or p-aminobenzyl acetal, etc., whose electronic structure is para-aminobenzylcarbamoyl. These include aromatic compounds similar to the PAB group.

[0127] Preferably, the self-immolative linker moiety has one of the following structures: [ka]

[0128] In the formula, ( * Atoms labeled with ) may be additional spacers or releasable linker units, or is the point of attachment to the cytotoxic agent and / or binding molecule (CBA); X 1 , Y 1 , Z 2 , and BiZ 3 are independently NH, O or S; Z 1 are independently H, NHR1, OR1, and SR1 , COX1R1, where X1 and R1 are as defined above; v is 0 or 1; U 1 are independently H, OH, C1-C6 alkyl, (OCH2CH2) n , F, Cl, Br , I, OR5, SR5, NR5R 5’ , N=NR5, N=R5, NR5R 5’ , NO2, SOR5R 5’ , SO2R5, SO3R5, OSO3R5, PR5R 5’ , POR5R5 ’ , PO2R5R 5’ , OPO(OR5)(OR 5’ ), or OCH2PO(OR5)( OR 5’ ) wherein R5 and R5 ’ are independently H, C1-C8 alkyl; C2-C 8 alkenyl, alkynyl, heteroalkyl, or amino acid; C3-C8 aryl, hetero ring, carbocycle, cycloalkyl, heterocycloalkyl, heteroaralkyl, alkylcarbocycle Bonyl, or glycoside; or a medicinal cationic salt;

[0129] W, L1, L2, V1, and V2 are independently non-self-immolative having one of the following structures: It may also contain a sexual linker component. [ka] JPEG0007637431000032.jpg180167

[0130] In the formula, ( * Atoms labeled with ) may be additional spacers or releasable linker units, or is the point of attachment to the cytotoxic agent and / or binding molecule; X 1 , Y 1 , U 1 , R5 and R5 ’ is defined as above; r is 0 to 100; m and n are independently 0 to 20. ru;

[0131] More preferably, W, L1, L2, V1, and V2 are independently releasable linker moieties. The term releasable refers to pH-labile, acid-labile, base-labile, oxidatively unstable cleavage under physiological conditions, such as a labile, metabolically unstable, biochemically unstable, or enzymatically labile bond refers to a linker containing at least one bond that can be broken. Chemical conditions do not necessarily include biological or metabolic processes, but instead include hydrolysis or substitution. Standard chemical reactions such as reactions in endosomes, which have a lower pH than the cytoplasmic pH, and / or dissociation with intracellular thiols such as glutathione, which are abundant in the millimolar range within malignant cells. It is understood that sulfide bond exchange reactions may be involved.

[0132] Examples of releasable linker moieties for W, L1, L2, V1, and V2 independently include: These include, but are not limited to: -(CR5R6) m (Aa) r (CR7R8) n (OCH2CH2) t -,-(CR5 R6) m (CR7R8) n (Aa) r (OCH2CH2) t -, -(Aa) r -(CR5 R6) m (CR7R8) n (OCH2CH2) t -, -(CR5R6) m (CR7R8) n (OCH2CH2) r (Aa) t -, -(CR5R6) m (CR7=R8)(CR9R 10 ) n (Aa) t (OCH2CH2) r -, -(CR5R6) m (NR 11 CO)(A a) t (CR9R 10 ) n (OCH2CH2) r -, -(CR5R6) m (Aa) t (N R 11 CO)(CR9R 10 ) n (OCH2CH2) r -, -(CR5R6) m (OCO )(Aa) t (CR9R 10 ) n (OCH2CH2) r -, -(CR5R6) m (OCN R7)(Aa) t (CR9R 10) n (OCH2CH2) r -, -(CR5R6) m (C O)(Aa) t -(CR9R 10 ) n (OCH2CH2) r -, -(CR5R6) m (N R 11 CO)(Aa) t (CR9R 10 ) n (OCH2CH2) r -, -(CR5R6) m (OCO)(Aa) t (CR9R 10 ) n -(OCH2CH2) r -,-(CR5R6 ) m (OCNR7)(Aa) t (CR9R 10 ) n (OCH2CH2) r -,-(CR5 R6) m (CO)(Aa) t (CR9R 10 ) n (OCH2CH2) r -,-(CR5R 6) m -phenyl-(CO)(Aa) t (CR7R8) n -, -(CR5R6) m -pretend Ru-(CO)(Aa) t (CR7R8) n -, -(CR5R6) m -oxazolyl-(C O)(Aa) t (CR7R8) n -, -(CR5R6) m -thiazolyl-(CO)(Aa ) t (CCR7R8) n -, -(CR5R6) t -Thienyl-(CO)(CR7R8) n -, -(CR5R6) t -Imidazolyl-(CO)(CR7R8) n -,-(CR5R6 ) t -morpholino-(CO)(Aa) t (CR7R8) n -, -(CR5R6) t -Pipe Ladino-(CO)(Aa) t (CR7R8) n -, -(CR5R6) t -N-Methylpiperidine Razine-(CO)(Aa) t (CR7R8) n -, -(CR5R) m -(Aa) t Feni Lu-,-(CR5R6) m -(Aa) t Frill-,-(CR5R6) m -oxazolyl( Aa) t -, -(CR5R6) m -Thiazolyl (Aa) t -, -(CR5R6) m -Chie Nil(Aa) t -, -(CR5R6) m -Imidazolyl (Aa) t -, -(CR5R6) m -Morpholino-(Aa) t -, -(CR5R6) m -Piperazino-(Aa) t -,-( CR5R6) m -N-methylpiperazino-(Aa) t -, -K(CR5R6) m (Aa) r (CR7R8) n (OCH2CH2) t -, -K(CR5R6) m (CR7R8) n ( Aa) r (OCH2CH2) t-、-K(Aa) r (CR5R6) m (CR7R8) n ( OCH2CH2) t -、-K(CR5R6) m (CR7R8) n (OCH2CH2) r ( Aa) t -、-K(CR5R6) m (CR7=R8)(CR9R 10 ) n (Aa) t (O CH2CH2) r -、-K(CR5R6) m (NR 11 CO)(Aa) t (CR9R 10 ) n (OCH2CH2) r -、-K(CR5R6) m (Aa) t (NR 11 CO)(CR 9R 10 ) n (OCH2CH2) r -、-K(CR5R6) m (OCO)(Aa) t (C R9R 10 ) n (OCH2CH2) r -、-K(CR5R6) m (OCNR7)(Aa) t (CR9R 10 ) n (OCH2CH2) r -、-K(CR5R6) m (CO)(Aa) t (CR9R 10 ) n (OCH2CH2) r -、-K(CR5R6) m (NR 11 CO) (Aa) t (CR9R 10 )n (OCH2CH2) r -, -K(CR5R6) m (OCO )(Aa) t (CR9R 10 ) n (OCH2CH2) r -, -K(CR5R6) m (OC NR7)(Aa) t (CR9R 10 ) n (OCH2CH2) r -, -K(CR5R6) m (CO)(Aa) t (CR9R 10 ) n (OCH2CH2) r -, -K(CR5R6) m -phenyl-(CO)(Aa) t (CR7R8) n -, -K(CR5R6) m -Frills- (CO)(Aa) t (CR7R8) n -, -K(CR5R6) m -oxazolyl-(CO )(Aa) t (CR7R8) n -, -K(CR5R6) m -thiazolyl-(CO)(Aa ) t (CR7R8) n -, -K(CR5R6) t -Thienyl-(CO)(CR7R8) n -, -K(CR5R6) t -Imidazolyl-(CO)(CR7R8) n -, -K(CR5 R6) t -morpholino-(CO)(Aa) t (CR7R8) n -, -K(CR5R6) t -piperazino-(CO)(Aa) t (CR7R8)n -, -K(CR5R6) t -N-Me Tylpiperazine-(CO)(Aa) t (CR7R8) n -, -K(CR5R) m -(Aa ) t Phenyl-, -K(CR5R6) m -(Aa) t Frill-, -K(CR5R6) m - Oxazolyl (Aa) t -, -K(CR5R6) m -Thiazolyl (Aa) t -, -K(C R5R6) m -Thienyl (Aa) t -, -K(CR5R6) m -Imidazolyl (Aa) t -, -K(CR5R6) m -Morpholino-(Aa) t -, -K(CR5R6) m -Pipera Gino (Aa) t -, -K(CR5R6) m -N-methylpiperazino-(Aa) t ; during the ceremony , m, Aa, m, and n are as defined above; t and r are independently 0 to 100. R3, R4, R5, R6, R7, and R8 are independently H; halide; C1-C 8 alkyl; C2-C8 aryl, alkenyl, alkynyl, ether, ester, amino or amide, and one or more halides, CN, NR1R2, CF3, OR1, Lille, heterocycle, S(O)R1, SO2R1, -CO2H, -S O3H, -OR1, -CO2R1, -CONR1, -PO2R1R2, -PO3H, or optionally substituted with P(O)R1R2R3; K is NR1, -SS -, -C(=O)-, -C(=O)NH-, -C(=O)O-, -C=NH-O-, -C =N-NH-, -C(=O)NH-NH-, O, S, Se, B, Het (heterocycle or C3 ~C8 heteroaromatic ring), or a peptide containing 1 to 20 amino acids.

[0133] Furthermore, the components W, L1, L2, V1, and V2 may independently have any of the following hydrophilic structures: May contain: [ka]

[0134] During the ceremony, [ka] is the binding site;

[0135] X2, X3, X4, X5, or X6 are independently NH; NHNH; N(R3); N(R 3)N(R 3’ );O;S;C1-C6 alkyl;C2-C6 heteroalkyl, ar Arylcycloalkyl, heterocycloalkyl; C3-C8 aryl, Ar-alkyl; Heterocycle, carbocycle, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl , heteroaryl; or 1 to 8 amino acids, wherein R3 and R 3’ is German In particular, H; C1-C8 alkyl; C2-C8 heteroalkyl, alkylcycloalky alkyl, heterocycloalkyl; C3-C8 aryl, Ar-alkyl, heterocycle, carbocycle , cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl or an ester, ether, or amide having 1 to 8 carbon atoms; or a group of the formula (OCH2 CH2) por (OCH2CH(CH3)) p Polyethyleneoxy units (p is 0 to an integer number of about 1000), or a combination thereof.

[0136] More preferably, the moieties W, L1, L2, V1, and V2 are independently 1 to 6 carbon atoms. A straight chain alkyl having atoms, or the formula (OCH2CH2) p Polyethyleneoxy units (p = 1 to 5000), or 1 to 4 units of amino acids (L or D form), or a combination of the above It is a combination.

[0137] Or, any one or more of W, Q1, Q2, L1, L2, V1, or V2 are independently Although they may not exist, Q1 and Q2 cannot exist simultaneously.

[0138] Generally, in another aspect, when V1 and / or V2 are linked to a cell-binding molecule T: Or, when L1 and / or L2 are directly bound to T (V1 and V2 are absent), , may be one or more of the following linker structures: [ka] JPEG0007637431000036.jpg238167JPEG0007637431000037.jpg41139

[0139] In the formula, R 20 and R 21 are independently C1-C8 alkyl; C2-C8 heteroalkyl or heterocycle; C3-C8 aryl, Ar-alkyl, cycloalkyl, alkylcyclo heterocycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, carbocycle, or Alkylcarbonyl; or (CH2CH2O) p C2~C with the formula 100 Polyethylene It is glycol (where p is defined as above).

[0140] In another further aspect, Q1 and Q2 are preferably C2 to C 18 Lipids, or C2 to C 18 Fatty acids, or C2-C 18 Polyalkylene glycol containing fatty ammonium lipids The polyalkylene glycol chains are selected to make the conjugate more hydrophilic during preparation. In addition to assisting the conjugate linker in the synthesis of hydrolytic enzymes, such as proteinases or esterases, Lipids protect the conjugates from being hydrolyzed by enzymes in mammalian blood. This can help bind the conjugate to the erythrocyte and slowly dissociate it from this complex during circulation. Thus, the side chain linkers of the present patent application make the conjugate more stable in circulation. Real alkylene glycols include poly(ethylene glycol) (PEG), poly(propylene glycol) glycol), and copolymers of ethylene oxide and propylene oxide. Particularly preferred is PEG, and even more particularly preferred is monofunctional PEG. Potentially activated hydroxyPEG (e.g., hydroxyPEG-monocarboxylic acid, HydroxyPEG-monoaldehyde, HydroxyPEG-monoamine, HydroxyPEG- Monohydrazide, HydroxyPEG-Monocarbazate, Reactive Ester of Hydroxyl Single-terminated activated hydroxyl PEG, including PEG-monoiodoacetamide , Hydroxyl PEG-monomaleimide, Hydroxyl PEG-monoorthopyridyldisil Sulfide, HydroxyPEG-monoxime, HydroxyPEG-monophenylcarbonate hydroxyl PEG-monophenylglyoxal, hydroxyl PEG-monothiophene Azolidine-2-thione, Hydroxyl PEG-monothioester, Hydroxyl PEG -monothiol, hydroxyl PEG-monotriazine, and hydroxyl PEG-mono The polyalkylene glycol is from about 10 Da to about 200 kDa. , preferably having a molecular weight of about 88 Da to about 40 kDa; More preferably, each of the two branches has a molecular weight of about 88 Da to about 40 kDa. In one particular embodiment, the polyalkylene has a molecular weight of about 88 Da to about 20 kDa. The glycol is poly(ethylene) glycol and is about 10 kDa; about 20 kDa; or In certain embodiments, the PEG has a molecular weight of about 40 kDa. PEG 20kDa (linear or branched), or PEG 40kDa (linear or branched). Linear or branched "non-antigenic" PEG polymers and their derivatives or conjugates The synthesis of the compound is described in many US patents, e.g., US Pat. Nos. 5,428,128; 5,621,033; 9;5,622,986;5,643,575;5,728,560;5,730,99 0;5,738,846;5,811,076;5,824,701;5,840,90 0;5,880,131;5,900,402;5,902,588;5,919,45 5;5,951,974;5,965,119;5,965,566;5,969,04 0;5,981,709;6,011,042;6,042,822;6,113,90 6;6,127,355;6,132,713;6,177,087; and 6,180, It is disclosed in 095.

[0141] Formula (I) or a pharmaceutically acceptable salt, hydrate, or hydrated salt thereof, or Polymorphic crystalline structures of compounds; or their optical isomers, racemates, diastereomers, or Examples of enantiomers are shown below: [ka] JPEG0007637431000039.jpg229166JPEG0007637431000040.jpg254165JPEG0007637431000041.jpg225166JPEG00076374310 00042.jpg229169JPEG0007637431000043.jpg239161JPEG0007637431000044.jpg246168JPEG0007637431000045.jpg228168 JPEG0007637431000046.jpg248168JPEG0007637431000047.jpg234167JPEG0007637431000048.jpg243168JPEG00076374310 00049.jpg220157JPEG0007637431000050.jpg235167JPEG0007637431000051.jpg228166JPEG0007637431000052.jpg235166

[0142] In the formula, Z 3 and Z 3 are independently H, OH, NH2, O, NH, COOH, COO, C (O), C(O), C(O)NH, C(O)NH2, R 18 , OCH2OP(O)(OR 18 )2, OC(O)OP(O)(OR 18 )2, OPO(OR 18 )2, NHPO(O R 18 )2, OP(O)(OR 18 )OP(O)(OR 18 )2, OC(O)R 18 , O C(O)NHR 18 , OSO2(OR18 ), O-(C4~C 12 -glycoside), linear or branched alkyl or heteroalkyl; C2-C8 straight or branched alkenyl, alkynyl alkyl, alkylcycloalkyl, heterocycloalkyl; C3-C8 linear or branched alkyl Ar, Ar-alkyl, heterocycle, carbocycle, cycloalkyl, heteroalkylcycloalkyl , alkylcarbonyl, heteroaryl; carbonate (-C(O)OR 17 ), Kalba Mate (-C(O)NR 17 R 18 ); or a molecular weight of about 88 daltons to about 20 kDa a polyalkylene glycol having the formula:

[0143] R 17 and R 18 are independently H, linear or branched alkyl or heteroalkyl; C C8 linear or branched alkenyl, alkynyl, alkylcycloalkyl, heterocycloa Aryl; C3-C8 linear or branched aryl, Ar-alkyl, heterocycle, carbocycle, cyclo Alkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl; Carbonate (-C(O)OR 17 ), carbamates (-C(O)NR 17 R 18 ) is;

[0144] R 19 are H, OH, NH2, OSO2 (OR 18 ), XCH2OP(O)(OR 18 ) 2. XPO(OR 18 )2, XC(O)OP(O)(OR 18 )2, XC(O)R 18 , XC(O)NHR 18 , C1-C8 alkyl or carboxylate; C2-C8 alkenyl aryl, alkynyl, alkylcycloalkyl, heterocycloalkyl; C3-C8 aryl or alkylcarbonyl; or medicinal salt;

[0145] X, X1, X2, and X3 are independently O, S, NH, NHNH, or CH2;

[0146] q1, q2, and q3 are independently selected from 0 to 24;

[0147] p, p1, and p2 are independently 1 to 100;

[0148] R 1’ , R 2’ , R 3’ , and R 4’ are independently selected from H and C1-C6 alkyl be;

[0149] Aa is a natural or unnatural amino acid; r is 0 to 12; if r>2, then (Aa) r is a peptide containing the same or different amino acid sequence; r=0 means that (Aa)r does not exist. It means not to;

[0150] m and n are independently 1 to 30.

[0151] In another embodiment of the present invention, the conjugate containing the side chain linkage is represented by formula (III): [ka]

[0152] In the formula, D, W, w, L1, L2, Q1, Q2, V1, V2, v1, v2, n, and T are the formulas This is the same definition as (I).

[0153] Formula (III) or a pharmaceutically acceptable salt, hydrate, or hydrated salt thereof, Polymorphic crystal structures of these compounds; or their optical isomers, racemates, diastereomers, or Examples of enantiomers are: [ka] JPEG0007637431000055.jpg244167JPEG0007637431000056.jpg217168JPEG00076374310 00057.jpg227168JPEG0007637431000058.jpg232164JPEG0007637431000059.jpg248166

[0154] In the formula, mAb, X1, X2, X3, p1, p2, p3, q1, q2, (Aa) r ,m, and n is as above.

[0155] In another aspect of the present invention, the side chain binding compound is represented by formula (IV), and is a cell binding molecule T or modified can readily react with a decorated cell-binding molecule T to form a conjugate of formula (I): [ka]

[0156] In the formula, D, W, w, L1, L2, Q1, Q2, V1, V2, v1, v2, and n are represented by the formula (I) is the same definition as

[0157] Lv1 is a molecule that binds thiols, amines, carboxylic acids, selenols, and phenols to cell-binding molecules. , or a reactive group capable of reacting with a hydroxyl group. Lv1 is OH, F, Cl, Br, I; Nitrophenol; N-hydroxysuccinimide (NHS); Phenol; Dinitrophenol phenol; pentafluorophenol; tetrafluorophenol; difluorophenol monofluorophenol; pentachlorophenol; triflate; imidazole; Dichlorophenol; Tetrachlorophenol; 1-Hydroxybenzotriazole; Sylate;Mesylate;2-Ethyl-5-phenylisoxazolium-3'-sulfonate with self-formed or other anhydrides, such as acetic anhydride or formic anhydride anhydrides formed; or for peptide coupling reactions or for Mitsunobu reactions The condensation reagent is selected from intermediate molecules generated by the condensation reagent for the reaction of EDC ( N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide), DCC (dicyclohexyl N,N'-diisopropylcarbodiimide (DIC), N-Cyclohexyl-N'-(2-morpholinoethyl)carbodiimide meso-p-toluene CMC, or CME-CDI, 1,1'-carbonyldiimidazole (CDI), TBTU(O-(benzotriazol-1-yl)-N,N,N',N'- Tetramethyluronium tetrafluoroborate), N,N,N',N'-tetramethyl -O-(1H-benzotriazol-1-yl)uronium hexafluorophosphate (HBTU), (benzotriazol-1-yloxy)tris(dimethylamino)phos Benzonium hexafluorophosphate (BOP), (benzotriazol-1-yloxy) Tripyrrolidinophosphonium hexafluorophosphate (PyBOP), diethyl Cyanophosphonate (DEPC), chloro-N,N,N',N'-tetramethylformamide Midinium hexafluorophosphate, 1-[bis(dimethylamino)methylene]-1 H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide hexafluoro Phosphate (HATU), 1-[(dimethylamino)(morpholino)methylene]-1H -[1,2,3]triazolo[4,5-b]pyridin-1-ium-3-oxide hexa Fluorophosphate (HDMA), 2-chloro-1,3-dimethylimidazolidinium Hexafluorophosphate (CIP), chlorotripyrrolidinophosphonium hexafluoro Fluorophosphate (PyCloP), Fluoro-N,N,N',N'-bis(tetramethyl N,N,N',formamidinium hexafluorophosphate (BTFFH) N'-Tetramethyl-S-(1-oxido-2-pyridyl)thiuronium hexafluoro Phosphate, O-(2-oxo-1(2H)pyridyl)-N,N,N',N'-tetraphosphate Methylthiuronium tetrafluoroborate (TPTU), S-(1-oxido-2-pi Lysyl)-N,N,N',N'-tetramethylthiuronium tetrafluoroborate, O -[(ethoxycarbonyl)cyanomethyleneamino]-N,N,N',N'-tetramethyl Fluorocarbonyl hexafluorophosphate (HOTU), (1-cyano-2-ethoxy- 2-Oxoethylideneaminooxy)dimethylamino-morpholino-carbenium hexa Fluorophosphate (COMU), O-(benzotriazol-1-yl)-N,N, N',N'-bis(tetramethylene)uronium hexafluorophosphate (HBPy U), N-benzyl-N'-cyclohexylcarbodiimide (with or without polymer linkages) (No mention), Dipyrrolidino(N-succinimidyloxy)-carbenium hexafluoro Phosphate (HSPyU), chlorodipyrrolidinocarbenium hexafluorophosphine 2-chloro-1,3-dimethylimidazolinium tetrafluoroborate (PyCIU), CIB, (benzotriazol-1-yloxy)dipiperidinocarbenium Hexafluorophosphate (HBPipU), O-(6-chlorobenzotriazole- 1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate (T CTU), bromotris(dimethylamino)phosphonium hexafluorophosphate ( BroP), propylphosphonic anhydride (PPACA, T3P®), 2-mol Methylisocyanide (MEI), N,N,N',N'-tetramethyl-O-(N- Succinimidyl)uronium hexafluorophosphate (HSTU), 2-bromo- 1-Ethyl-pyridinium tetrafluoroborate (BEP), O-[(ethoxycarbonyl) N,N,N',N'-tetramethyluronium tetrafluoromethyl Fluoroborate (TOTU), 4-(4,6-dimethoxy-1,3,5-triazine-2 -yl)-4-methylmorpholinium chloride (MMTM, DMTMM), N,N,N' ,N'-Tetramethyl-O-(N-succinimidyl)uronium tetrafluoroborate TSTU, O-(3,4-dihydro-4-oxo-1,2,3-benzotriazine -3-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate ( TDBTU), 1,1'-(azodicarbonyl)dipiperidine (ADD), di-(4-chloro-2-methyl-2-pyridin-1-yl)-4-methyl-2-pyridin-1-yl Di-tert-butyl azodicarboxylate (DCAD), di-tert-butyl azodicarboxylate Diisopropyl azodicarboxylate (DBAD), diisopropyl azodicarboxylate (DIAD), diisopropyl azodicarboxylate (DIAD), Lv1 is selected from the group consisting of the acid itself and may be an anhydride formed with other C1-C8 anhydrides;

[0158] Preferred Lv1 are halides (e.g., fluorides, chlorides, bromides, and iodides). ), methanesulfonyl (mesyl), toluenesulfonyl (tosyl), trifluoromethyl Sulfonyl (triflate), trifluoromethyl sulfonate, nitrophenoxyl, N-Succinimidyloxyl (NHS), Phenoxyl; Dinitrophenoxyl; Penta Fluorophenoxyl, tetrafluorophenoxyl, trifluorophenoxyl, difluorophenoxyl Fluorophenoxyl, monofluorophenoxyl, pentachlorophenoxyl, 1H-isopropyl Midazol-1-yl, chlorophenoxyl, dichlorophenoxyl, trichlorophenoxyl Xyl, tetrachlorophenoxyl, N-(benzotriazol-yl)oxyl, 2- Ethyl-5-phenyl-3'-sulfonyl, phenyloxadiazole-sulfonyl(- Sulfone-ODA), 2-ethyl-5-phenylisoxazolium-yl, phenyl Oxadiazol-yl (ODA), oxadiazol-yl, unsaturated carbon (carbon-carbon, Carbon-nitrogen, carbon-sulfur, carbon-phosphorus, sulfur-nitrogen, phosphorus-nitrogen, oxygen-nitrogen, or carbon -oxygen double or triple bond), or one of the following structures:

[0159] [ka] TIFF0007637431000062.tif166167

[0160] In the formula, X 1’ is F, Cl, Br, I, or Lv3; X 2’ are O, NH, N(R1 ), or CH2; R3 is independently H, aromatic, heteroaromatic, or one or more H atoms are independently -R1, -halogen, -OR1, -SR1, -NR1R2, -NO2, an aromatic group substituted with -S(O)R1, -S(O)2R1, or -COOR1; Lv3: F, Cl, Br, I, nitrophenol; N-hydroxysuccinimide phenol; dinitrophenol; pentafluorophenol; tetrafluorophenol Difluorophenol; Monofluorophenol; Pentachlorophenol Triflate; Imidazole; Dichlorophenol; Tetrachlorophenol; 1-Hydroxybenzotriazole; Tosylate; Mesylate; 2-Ethyl-5-phenyl isoxazolium-3'-sulfonates, self-formed or other anhydrides, leaving groups selected from anhydrides formed with, for example, acetic anhydride or formic anhydride; or by condensation reagents for peptide coupling reactions or for Mitsunobu reactions. It is an intermediate molecule formed.

[0161] Formula (IV) or a pharmaceutically acceptable salt, hydrate, or hydrated salt thereof, or Polymorphic crystalline structures of the compounds; or their optical isomers, racemates, diastereomers, or Examples of enantiomers are shown below: [ka] TIFF0007637431000064.tif237164TIFF0007637431000065.tif236170TIFF0007637431000066.tif247162TIFF0007637431000067.t if222166TIFF0007637431000068.tif248160TIFF0007637431000069.tif234157TIFF0007637431000070.tif245168TIFF00076374310 00071.tif246170TIFF0007637431000072.tif229170TIFF0007637431000073.tif232158TIFF0007637431000074.tif223157TIFF000 7637431000075.tif229160TIFF0007637431000076.tif223163TIFF0007637431000077.tif234160TIFF0007637431000078.tif201158

[0162] In the formula, X1, X2, X3, Z2, Z3, p.p1, p2, p3, q1, q2, Lv3, (Aa) r , R 25 , R 25’ , and m are as above.

[0163] In another embodiment of the present invention, the side chain binding compound is represented by formula (V), and can be easily bound to a cell-binding molecule T. can react to form a conjugate of formula (III): [ka]

[0164] D, W, w, L1, L2, Q1, Q2, V1, V2, v1, v2, and n are each a group represented by formula (I). wherein Lv1 and Lv2 independently have the same definition as Lv1 in formula (IV). and in formula (V), both Lv1 and Lv2 may be the same or different.

[0165] Examples of formula (V) are shown below: [ka] TIFF0007637431000081.tif225156TIFF0007637431000082.tif234157TIFF00076374310 00083.tif228155TIFF0007637431000084.tif229158TIFF0007637431000085.tif192155

[0166] In the formula, X1, X2, X3, Z2, Z3, p, p1, p2, p3, q1, q2, Lv3, (Aa) r , R 25 , R 25’ , and m are as above.

[0167] The present invention further provides cell-binding molecule-tubulysin analog conjugates of formula (I) and formula (III) The present invention relates to a method for preparing the conjugates of formula (I) and formula (I) and to applications of the conjugates of formula (I).

[0168] The cell binding agent / molecule T is a molecule that binds to a residue of a cell population that is to be therapeutically or otherwise biologically modified. Any molecule now known or discovered that binds to, complexes with, or reacts with Preferably, the cell binding agent / molecule is an immunotherapeutic protein, antibody, single chain antibody, target cell Cell-binding antibody fragments, monoclonal antibodies, single-chain monoclonal antibodies, target cell binding monoclonal antibody fragments, chimeric antibodies, chimeric antibody fragments that bind to target cells, domain Antibodies, target cell-binding domain antibody fragments, antibody-mimicking adnectins, DARPs ins, lymphokines, hormones, vitamins, growth factors, colony-stimulating factors, nutrient transporters Attached to transferrin, and / or cell-binding peptides, proteins, and albumin small molecules, polymers, dendrimers, liposomes, nanoparticles, vesicles, or (viruses) capsids.

[0169] Preferred Lv1 and Lv2 react with thiol pairs of cell-binding agents / molecules. The alcohol is preferably dithiothreitol (DTT), dithioerythritol (DTE), or the like. , L-glutathione (GSH), tris(2-carboxyethyl)phosphine (TCEP ), 2-mercaptoethylamine (β-MEA), and / or β-mercaptoethanol ( The interchain disulfide bonds of the cell-binding molecules are reduced by a reducing agent selected from the group consisting of β-ME, 2-ME, and β-ME. The bond is a pair of sulfur atoms produced by reduction.

[0170] Preparation of conjugates of cell-binding molecules with tubulysin analogues via side chain linkages

[0171] Preparation of conjugates of tubulysin analogs to cell-binding molecules of the present invention and their side chain attachment The synthetic routes to produce the conjugates are shown in Figures 1-51.

[0172] The conjugates of formula (I) and (III) can be prepared by reacting intermediate compounds of formula (IV) and (V), respectively. Generally, the tubulysin analogs of formulas (IV) and (V) can be prepared via cell binding. Lv1 and Lv2 functionalities that can readily react with molecules or modified cell-binding molecules The tubulysin analogs of formulas (IV) and (V) are synthesized to have a group. Some preparations of formula (I) and (III) are structurally depicted in Figures 1-51.

[0173] The conjugate of formula (I) is generally prepared by subjecting the conjugate to a HCl solution containing DMA, DMF, Ethanol, methanol, acetone, acetonitrile, THF, isopropanol, di Water-miscible organic solvents such as hexane, propylene glycol, and ethylenediol With or without adding 0 to 30% of a solvent, the functional group Lv1 of formula (IV) and one of the cell-binding molecules, and then the compound of formula (I) formed is dialyzed or It can be synthesized by purifying it by chromatography. Some of the reactive groups for conjugation can be obtained through protein engineering.

[0174] The conjugate of formula (III) is linked to the functional group L of the linker of formula (V) to form the conjugate molecule. v1 and Lv2 and two or more residues of the cell-binding molecule, preferably at 0 to 60°C, Cells were collected in an aqueous medium of pH 5-9 with or without the addition of 100% water-miscible (miscible) organic solvent. Reaction with a pair of free thiols generated by reducing the disulfide bonds of the polymer The pair of free thiols can also be obtained by mixing with preferably 0 to 30% water. In an aqueous medium of pH 4-9 with or without the addition of a miscible organic solvent, dithiothiazol-1-one is used. DTT, dithioerythritol (DTE), L-glutathione (GSH) , tris(2-carboxyethyl)phosphine (TCEP), 2-mercaptoethylamine Select from β-MEA, and / or β-mercaptoethanol (β-ME, 2-ME) The interchain disulfide bonds of the cell-binding agent are reduced by a reducing agent capable of It is the opposite of conjunction.

[0175] The reactive groups Lv1 and Lv2 in formula (IV) and formula (V) are independently disulfides. , thiol, maleimide, halogen-substituted maleimide, haloacetyl, azide, 1-yne, Ketone, aldehyde, alkoxyamino, triflate, carbonylimidazole, toxin 2-Ethyl-5-phenylisoxazolium-3'-sulfonate, mesylate carboxylic acid esters of nitrophenols, N-hydroxysuccinimide (NH S), phenol; dinitrophenol, pentafluorophenol, tetrafluorophenol phenol, difluorophenol, monofluorophenol, pentachlorophenol, Dichlorophenol, tetrachlorophenol, 1-hydroxybenzotriazole, no It can be a hydrate, a hydrazide group, or other acid ester derivatives, and at 0 to 60°C, Aqueous solutions at pH 5-9 with or without the addition of 0-30% of water-soluble (miscible) organic solvents In a medium, one, two or more groups of cell-binding molecules / drugs are reacted with each other simultaneously or sequentially. can be reacted to produce the conjugates of formula (I) and formula (III) after column purification or dialysis. The reactive groups Lv1 and Lv2 in formula (IV) and formula (V) can be immediately reacted in different ways: React with modified cell-binding molecules, such as cell-binding agents of formula (I) - tubulysins The linkages containing disulfide bonds in the homologous conjugates are preferably disulfide bonds in the modified cell-binding agent. By disulfide exchange between the thiol bond and Lv1 and Lv2, which have free thiol groups, or the free thiol groups of the modified cell-binding agent and the disulfides on Lv1 and / or Lv2 This is achieved by disulfide exchange between the bonds. Therefore, the disulfide group is usually replaced by disulfanylpyridine, disulfanyl-nitropyridine, disulfanyl-nitrobenzene, disulfanyl-nitrobenzoic acid, or disulfanyl The thio groups in the conjugates of formula (I) and formula (III) are phenyl-dinitrobenzene and the like. The linkages containing ether bonds can be attached to the modified cell-binding agent or to the tubes of formula (IV) and formula (V). -maleimide or haloacetyl or ethylsulfonyl on the glycine analogue and formula (IV) and a free thiol group on the tubulysin analogue of formula (V) or on the modified cell-binding agent, respectively. This is achieved by reaction with the hydroxyl group. In this case, the method may be performed using methods known in the art (e.g., P. Hamann et al., Cancer Res. 53, 33 36-34, 1993; B. Laguzza et al., J. Med. Chem., 32; 548-55, 1959; P. Trail et al. , Cancer Res., 57; 100-5, 1997), the drug or modified a carbonyl group on the cell-binding molecule and a carbonyl group on the drug or modified The inclusion of triazoles in the conjugates achieved by reaction with hydrazide residues on cell-binding molecules. The conjugation of the hydroxyl group containing the bond can be achieved via click chemistry (Huisgen cycloaddition) (Lutz, J.F. et al. al, 2008, Adv. Drug Del. Rev.60, 958-70; Sletten, EM et al 2011, AccChem. R Research 44, 666-76) on the drug or cell-binding molecule of formula (IV) and formula (V) This is achieved by reaction of the 1-yne group with the azide moiety of another counterpart. The bond containing the oxime bond in the conjugate linked via The ketone or aldehyde groups on the surface or on cell-associated molecules and the oxyl groups on the other counterparts This is achieved by reaction with amine groups. Thiol-containing cell-binding molecules are suitable for use at pH 5.5-9.0. in an aqueous buffer of the formula The thiol-containing cell-binding molecule can react with the drug molecule conjugate of formula (IV) and formula (V). The compound of formula (IV) and formula (V) having a lysyldithio residue undergoes disulfide exchange with the drug conjugates. By adding the hydroxyl group, a conjugate having a disulfide bond can be obtained. Alternatively, a cell-binding molecule having a thiol group may be bound under mildly basic conditions, for example, at pH 8.0 to 9.5. and the formula (IV) and the formula (V) with halogen, in particular alpha halide of carboxylate. ) a modified drug having an ether or thioether bond that can react with a drug conjugate of The hydroxyl or amino groups of the cell-binding molecules can be dehydrated by dehydration with EDC or DCC. In the presence of an agent, it can be condensed with a drug conjugate of formula (IV) or (V) having a carboxyl group, The cell-binding molecule containing an amino group can be linked to the drug of formula (IV) or (V) NHS, imidazole, nitrophenol, and N-hydroxysuccinimide on the linker Carboxyl ester group; Phenol; Dinitrophenol; Pentafluorophenol ;Tetrafluorophenol;Difluorophenol;Monofluorophenol;Penta Chlorophenol; Triflate; Imidazole; Dichlorophenol; Tetrachlorophenol phenol; 1-hydroxybenzotriazole; tosylate; mesylate; or 2-ethyl It can be condensed with 5-phenylisoxazolium-3'-sulfonate, A conjugate is obtained via a bond.

[0176] The synthesized conjugates can be purified by standard biochemical methods, e.g., by chromatography on Sephadex G25 or S Gel filtration using ephacryl S300 columns, adsorption chromatography, and ion exchange chromatography In some cases, small molecules can be purified by exchange or dialysis. Conjugation of molecules (e.g., folic acid, melanocyte-stimulating hormone, EGF, etc.) with small molecule drugs In this case, chromatography, such as HPLC, medium pressure column chromatography, or ion chromatography may be used. It can be purified by ion exchange chromatography.

[0177] A pair of free thiol pairs on the cell-binding molecule, preferably an antibody, and a compound represented by formula (I) or (III) To achieve higher conjugation yields with HCl, a small amount of water-miscible organic solvent or phase inversion solvent may be added to the reaction mixture. First, a cross-linking agent of formula (IV) or formula (V) ( The linker is dissolved in a water-miscible polar organic solvent, such as methanol, ethanol, or propanolic acid. different alcohols such as acetone, acetonitrile, tetrahydrofuran (THF), 1,4-dioxane, dimethylformamide (DMF), dimethylacetamide (DM A) or dimethyl sulfoxide (DMSO) at a high concentration, for example, 1 to 500 mM. On the other hand, it can be concentrated in an aqueous buffer solution of pH 4 to 9.5, preferably 6 to 8.5. For cell-binding molecules such as antibodies dissolved at 1 to 50 mg / ml, 0.5 to 20 equivalents of TCEP or DTT for 20 minutes to 48 hours. After reduction, SEC chromatography purification is performed. TCEP can also be removed by SEC chromatography, if desired. Alternatively, the reaction mixture can be used for the next step reaction without purification. TCEP can be stored in 4-azidobenzoic acid, 4-(azidomethyl)benzoic acid, benzoic acid, or azido-polyethylene glycolyl (e.g., 2-(2-(2-(2-azido It is preferable to neutralize with an azide compound such as ethoxy, ethoxy, ethoxyethanol, etc. Furthermore, in order to achieve cross-linking of cell-binding molecules simultaneously with TCEP reduction, The reduction of an antibody or other cell-binding agent by the compound of formula (IV) or formula (V) results in a drug conjugate molecule of formula (IV) or formula (V). This can be done with the presence of

[0178] The aqueous solution for the modification of cell-binding agents has a pH between 4 and 9, preferably between 6.0 and 7.5. It can be buffered and contain non-nucleophilic buffer salts useful in these pH ranges. Agents include phosphate, acetate, triethanolamine HCl, HEPES, and MOP. S buffers, and further, for example, cyclodextrin, hydroxypropyl-β-cyclodextrin rhodextrin, polyethylene glycol, sucrose, and salts (e.g., NaCl, K In a solution containing reduced cell-binding molecules, an additional component such as HCl may be present. After adding the drug conjugate of formula (IV) or (V), the reaction mixture is heated at 4°C to 45°C, preferably The reaction is incubated at ambient temperature of 15°C. The progress of the reaction is monitored by the decrease in absorbance at 254 nm. Measure the increase in absorbance at 280 nm or 280 nm, or at other suitable wavelengths. After the reaction is complete, isolation of the modified cell-binding agent can usually be performed by methods for the preparation of chromatographically active compounds, such as gel filtration chromatography, ion exchange chromatography, adsorption chromatography, or column chromatography on silica gel or alumina; Crystallization, preparative thin layer chromatography, ion exchange chromatography, or HPLC It can be done more.

[0179] The degree of modification can be determined by the UV spectrum emitted by nitropyridinethiones, dinitropyridines, and Lysine dithione, pyridine thione, carboxyamidopyridine dithione, and dicarboxy The development can be evaluated by measuring the absorbance of the thiamidopyridinedithione group. For conjugates without a chromophore group, the modification or conjugation reaction can be monitored by LC-MS, preferably UPL. Monitored by C-QTOF mass spectrometry or capillary electrophoresis (CEMS) The side chain bridges described herein can be used with any drug with appropriate substituents. The hydroxyl groups have a variety of functional groups that can react with a cytotoxic agent, preferably a cytotoxic agent. For example, amino or hydroxyl groups Modified cell-binding molecules with hydroxy substituents are N-hydroxysuccinimide (NHS) esters. Modified cell-binding molecules with thiol substituents can react with drugs bearing esters. It can react with drugs that have maleimide or haloacetyl groups. Modified cell-binding molecules with alkyl substituents (ketones or aldehydes) are The drug can react with a hydroxylamine-containing compound. The linker to be used can be readily determined based on the known reactivity of the functional groups.

[0180] Cell-binding agents

[0181] The cell-binding molecule Cb constituting the conjugate and modified cell-binding molecule of the present invention can be used therapeutically. or other biologically modified residues of the cell population to be modified. It may be any molecule that is currently known or that will be discovered.

[0182] Cell binding agents include, but are not limited to, large molecular weight proteins, such as full-length antibodies (PPs). Reclonal or monoclonal), dimeric, multimeric, multispecific antibodies (e.g., bispecific single-chain antibodies; antibody fragments, e.g., Fab, Fab', F(ab')2, Fv [Parham, J. Immunol. 131, 2895-2902 (1983)], F Fragments obtained by ab expression libraries, anti-idiotypic (anti-Id) antibodies, CDRs, bispecific antibodies, trispecific antibodies, cancer cell antigens, viral antigens, microbial antigens, or The immunoglobulin is capable of recognizing and binding to a specific antigen or exhibiting a desired biological activity. epitope-binding fragments of any of the foregoing that immunospecifically bind to a protein produced in an immune system; Interferons (e.g., types I, II, III); peptides; lymphokines, e.g., IL-2, IL-3, IL-4, IL-5, IL-6, IL-10, GM-CSF, or Interferon gamma (IFN-γ); hormones, e.g., insulin, TRH (thyroid stimulator estrogen-releasing hormone), MSH (metastatic hormone), or androgen, estrogen steroid hormones such as steroids (e.g., steroid hormones) or melanocyte-stimulating hormone (MSH); growth factors and and colony-stimulating factors, e.g., epidermal growth factor (EFG), granulocyte-macrophage colony-stimulating factors stimulating factors (GM-CSF); transforming growth factors (TGF), e.g., TGF α, TGFβ; insulin and insulin-like growth factors (IGF-I, IGF-II) G -CSF, M-CSF, and GM-CSF [Burgess, Immunology T Today, 5, 155-158 (1984)]; Vaccine Growth Factor (VGF); Fibroblast Growth Factor (FGF) Follicular growth factors (FGFs); small molecular weight proteins, polypeptides, peptides, and peptide phosphatase inhibitors steroids, such as bombesin, gastrin, and gastrin-releasing peptide; platelet-derived Growth factors; interleukins and cytokines, e.g., interleukin-2 (IL-2 ), interleukin-6 (IL-6), leukemia inhibitory factor, granulocyte-macrophage colony -stimulating factor (GM-CSF); vitamins such as folic acid; apoproteins and glycoproteins, e.g. For example, transferrin [O'Keefe et al, J. Bio. Chem. 260 , 932-927 (1985)]; carbohydrate-binding proteins such as lectins or lipoproteins; vesicle nutrient transport molecules; and small molecule inhibitors, e.g., inhibition of prostate-specific membrane antigen (PSMA) drugs, small molecule tyrosine kinase inhibitors (TKIs), non-peptides, or other cell-binding molecules or materials, such as bioactive polymers (Dhar, et al., Proc. Natl. A cad.Sci.2008,105,17356-61), bioactive dendrimers (Le e,et al,Nat.Biotechnol.2005,23,1517-26;A lmutairi,et al;Proc.Natl.Acad.Sci.2009,1 06,685-90), nanoparticles (Liong, et al, ACS Nano, 200 8,19,1309-12;Medarova,et al,Nat.Med.2007 ,13,372-7;Javier,et al,Bioconjugate Chem .2008,19,1309-12), liposomes (Medinai,et al,Cu rr. Phar. Des. 2004, 10, 2981-9), viral capsid (Fle nniken,et al,Viruses Nanotechnol.2009,32 7,71-93).

[0183] Generally, if a suitable monoclonal antibody is available, it can be used to target the cell surface. Antibodies are preferred as binding molecules. Antibodies may be murine, human, humanized, chimeric, or derived from other species. The antibodies used in the present invention may be produced in vivo or in vitro. The process for producing anti-receptor peptide polyclonal antibodies includes the process for producing anti-receptor peptide polyclonal antibodies. Methods for preparing the compound are shown, for example, in U.S. Pat. No. 4,493,795 (Nestor et al.). The typical method for preparing monoclonal antibodies is by immunization with a specific antigen. This method involves fusing mouse spleen cells isolated from mice with myeloma cells (K ohler, G; Milstein, C. 1975. Nature 256:495-4 97) For detailed operating instructions, see Antibodies-A Laboratory Manual,Harlow and Lane,eds.,cold spring Harbor Laboratory Press, New York (1988) The contents of that document are incorporated herein by reference. In particular, methods for immunizing mice, rats, hamsters, or other mammals with an antigen of interest. By this method, monoclonal antibodies can be obtained, and the target antigen can be, for example, an intact target cells, antigens isolated from target cells, whole viruses, weakened whole viruses and viruses Examples of the protein include PEG6000, which is used to fuse spleen cells with myeloma cells. The hybridomas obtained after fusion were screened using their sensitivity to HAT. Hybridomas producing monoclonal antibodies useful in the practice of the present invention are , identified by suppressing immune reactions or receptor activity with specific target cell receptors. .

[0184] The monoclonal antibodies used in the present invention are those secreting antibodies with appropriate antigen specificity. Start a monoclonal hybridoma cell culture in a nutrient medium containing hybridoma cells. In the culture, the hybridoma cells release the antibody into the culture medium. The antibody-containing culture supernatant was collected and the conditions and time required for the antibody to be secreted into the culture medium were maintained. This is followed by well-known techniques, such as protein A affinity chromatography, anion exchange chromatography, etc. Chromatography, cation exchange chromatography, hydrophobic interaction chromatography and molecular sieve chromatography (especially affinity chromatography using antigen-crosslinking protein A) chromatography and molecular sieve chromatography), centrifugation, sedimentation, or other methods. The antibodies can be isolated by standard methods for protein purification.

[0185] Media useful for the preparation of these compositions are well known in the art and commercially available. Exemplary synthetic media include Dulbecco's Minimum Essential Medium (MIM) DMEM (Dulbecco et al., Virol. 8:396 (1959)), 4.5 g m / L glucose, 0-20mM glutamine, 0-20% fetal bovine serum, ppm Some heavy metals (e.g., Cu, Mn, Fe, or Zn) or / and added in salt form Heavy metals, and antifoaming agents (e.g., polyoxyethylene-polyoxypropylene block copolymers) Polymer) is added.

[0186] In addition to cell fusion technology, we have also constructed cell lines for antibody production using the following methods: For example, direct transformation of B lymphocytes by oncogenic DNA can or oncogenic viruses, such as Epstein-Barr virus (EBV, human herpes simplex virus) Also known as pesvirus 4 (HHV-4) or Kaposi's sarcoma-associated virus Transfection with KSHV (see U.S. Patent No. 4,341,761 for details) ;4399121;4427783;4444887;4451570;4466917 ;4472500;4491632;4493890). Monoclonal antibodies are Based on known methods, anti-receptor peptides or peptides containing terminal carboxyl groups can be used. can be prepared (see Niman et al. Proc. Natl. Acad. S ci.USA,80:4949-4953(1983);Geysen et al. Proc.N atl.Acad.Sci.USA,82:178-182(1985); Lei et al. B Biochemistry 34(20):6675-6688(1995)). Typically, the anti-receptor polypeptide or polypeptide analog is an anti-receptor polypeptide of a monoclonal antibody. Use as an immunogen to prepare polypeptides, either alone or in a cross-linked immunogenic carrier It is possible.

[0187] To produce monoclonal antibodies as binding molecules of the present invention, other well-known production methods can be used. Among them, the method of producing fully human antibodies has attracted particular attention. Display technology uses affinity selection to select antibodies specific to a known antigen from a fully human antibody library. Phage display technology itself is widely used in the literature to obtain fully human antibodies that bind to specific antigens. , vector construction, and library screening are described in detail. Dente et al. Gene. 148(1):7-13 (1994); Little et al. Biotechnol Adv.12(3):539-55(1994);Clacks on et al. Nature 352:264-628(1991); Huse et al. Scien See ce 246:1275-1281 (1989).

[0188] Monoclonal antibodies derived from non-human species (e.g., mice) using hybridoma technology For cloned antibodies, humanization is performed to avoid human anti-mouse antibodies when administered to humans. Among these, the well-known method for antibody humanization is the method of humanizing the complementarity-determining region (CDR) of the antibody. For more information, see U.S. Patent Nos. 5,859,205 and 6,859,205. , No. 797, 492; Liu et al., Immunol Rev. 222:9-27 (2008 );Almagro et al., Front Biosci.1;13:1619-33(200 8);Lazar et al., Mol Immunol.44(8):1986-98(2007 );Li et al. Proc.Natl.Acad.Sci.USA.103(10):355 7-62 (2006), the disclosure of which is incorporated by reference. The transgenic mouse, U, possesses most of the human immunoglobulin light and heavy chains. It can be prepared by immunizing herons, monkeys, and other mammals with an antigen. Examples of such mice include Xenomouse (Abgenix, Inc.), HuM ab-Mouse(Medarex / BMS), VelociMouse(Regene For details, see U.S. Patent Nos. 6,596,541, 6,207,418, No. 6,150,584, No. 6,111,166, No. 6,075,181, No. 5,922,5 Nos. 45, 5,661,016, 5,545,806, 5,436,149 and 5, See US Pat. No. 569,825. In the course of human therapy, mouse antibody variable region genes and human antibody The immunogenicity of chimeric antibodies produced in the human body by integrating the human constant region genes is Much lower than mouse antibodies (Kipriyanov et al., Mol Biotech nol.26:39-60(2004);Houdebine,Curr Opin B Biotechnol. 13:625-9 (2002)). The disclosure of said document is incorporated by reference. Furthermore, site-directed mutagenesis of antibody variable regions can be used to improve the quality of antibody parental sequences. This can improve compatibility and specificity (Brannigan et al., Nat Rev M ol Cell Biol.3:964-70(2002);Adams et al., J.Imm Univ. Methods. 231:249-60 (1999)). By replacing the cytotoxicity of the cytotoxicity domain with the cytotoxicity of the immune effector cells, the cytotoxicity of the immune effector cells can be effectively promoted. The toxic effect can be enhanced.

[0189] Immunospecific antibodies against malignant cell antigens can be obtained commercially or by several conventional techniques. For example, they can be obtained by chemical synthesis or recombinant expression techniques. Nucleotide sequences encoding immunospecific antibodies to Other similar databases, commercial sources, published literature, or routine cloning and It can be obtained by sequencing.

[0190] Besides antibodies, polypeptides or proteins can also be used as binding molecules to target cell surfaces. Interact with the corresponding receptor or epitope by binding, blocking, attacking, or other means These peptides or proteins bind to epitopes or their corresponding receptors. They do not have to belong to the immunoglobulin family, as long as they can bind effectively. The polypeptides are also isolated by a technique similar to that of phage display antibodies (Sza et al., rdenings,J Recept Signal Transduct Res.2 003;23(4):307-49). Peptides obtained from a random peptide library The polypeptide or protein fragments are similar in application to antibodies and antibody fragments. The molecule binds to the antigen by connecting to some macromolecule or medium via a binding molecule. These macromolecules include, but are not limited to, albumin, polymers, , liposomes, nanoparticles, or dendrimers.

[0191] Drugs based on the conjugates of the present invention for treating cancer, autoimmune diseases, and infectious diseases Antibodies used for binding include, but are not limited to, 3F8 (antibody GD2 antibody), abagovomab (anti-CA-125 antibody), abciximab (anti-CD41 antibody) (integrin α-IIb)), adalimumab (anti-TNF-α antibody), adalimumab (anti EpCAM antibody, CD326), afelimomab (anti-TNF-α); afutuzumab (anti-C D20 antibody), Alacizumab pegol (anti-VEGFR2 antibody), ALD518 (anti-IL-6 antibody), alemtuzumab (also known as Campus, MabCampus) , anti-CD52 antibody), altumomab (anti-CEA antibody), anatumomab (anti-tag-72 antibody) body), anrukinzumab (IMA-638, anti-IL-13 antibody), apolizumab (anti-H LA-DR antibody), arcitumomab (anti-CEA antibody), acelizumab (anti-L-selectin (CD62L) antibody), Atlizumab (also known as tocilizumab, Actemra , RoActemra, anti-IL-6 receptor antibody), Atorolimumab (anti-albuminuria) Ghesal factor antibody), bapineuzumab (anti-β-amyloid antibody), basiliximab (simuletin) anti-CD25 (IL-2 receptor α chain) antibody), bavituximab (anti Phosphatidylserine antibody), Bectumomab (also known as LymphoScan, anti-CD 22 antibody), belimumab (also known as BENLYSTA, LymphoStat-B, anti-BAFF antibody), benralli Benralizumab (anti-CD125 antibody), bertilimumab (anti-CCL11 (eosinophil) Scintimun-1 antibody), besilesomab (also known as Scintimun, anti-CEA-related antigen antibody), beba Ignitumab (also known as Avastin, an anti-VEGF antibody), biciromab (also known as FibriScint, an anti-fibroblast growth factor antibody) Bivatuzumab (anti-CD44v6 antibody), blinatumomab (blin atumomab) (also known as BiTE, anti-CD19 antibody), brentuximab ( cAC10, anti-CD30 TNFRSF8 antibody), Briakinumab (anti- IL-12, IL-23 antibody), canakinumab (also known as Ilaris, anti-IL-1 antibody) , cantuzumab (also known as C242, anti-CanAg antibody), capromab, Maxomab (also known as removab, anti-EpCAM, anti-CD3 antibody), CC49 (anti-TAG-7 2 antibody), cedelizumab (anti-CD4 antibody), certolizumab pegol (another CIMZIA, anti-TNF-α antibody), cetuximab (also known as Erbitux, IMC-C22 5, anti-EGFR antibody), sitatuzumab (anti-EpCAM antibody), cixutumbam mab) (anti-IGF-1 antibody), clenoliximab (anti-CD4 antibody), clivatuzumab (Cli vatuzumab) (anti-MUC1 antibody), conatumumab (anti-TRAIL-R2 antibody), CR6261 (anti-influenza A hemagglutinin antibody), dacetuzumab zumab) (anti-CD40 antibody), daclizumab (also known as Zenapax, anti-CD25C (IL-2 receptor) Anti-CD38 (cyclic ADP) antibody), Daratumumab (anti-CD38 (cyclic ADP) ribose hydrolase antibody), denosumab (also known as Prolia, anti-RANKL antibody), Momab (anti-B-lymphoma cell antibody), dorlimomab, dorlixizumab, Ecromeximab (anti-GD3 ganglioside antibody), eculizumab (another Name: Soliris (anti-C5 antibody), edovacomab (anti-endotoxin antibody), edrecolomab (Also known as Panorex, MAb17-A1, anti-EpCAM antibody), efalizumab (Also known as Rap tiva, anti-LFA-1 (CD11a) antibody), efungumab (also known as Mycog rab, anti-Hsp90 antibody), elotuzumab (anti-SLAMF7 antibody), Elsilimomab (anti-IL-6 antibody), enlimomab pegol (anti-ICAM-1 (CD54) antibody), Epitumomab (anti-episialin antibody), Epratuz Mab (anti-CD22 antibody), Erlizumab (anti-ITGB2 (CD18) antibody) ), Ertumaxomab (also known as Rexomun, anti-HER2 / neu, anti-CD3 body), etaracizumab (also known as Abegrin, anti-integrin αvβ3), exibirmab (anti-hepatitis B surface antigen antibody (HBs antibody)), Fanolesomab (also known as N eutroSpec, anti-CD15 antibody), faralimomab antibody (anti-interferon Farletuzumab (anti-folate receptor 1 antibody), Farbi Felvizumab (antibody against respiratory syncytial virus), Fezakinumab (anti-IL-22 antibody), Figitumumab (anti-IGF-1 receptor antibody), Fontolizumab (anti-IFN-γ antibody), foravirumab b) (anti-rabies virus glycoprotein antibody), fresolimumab (anti-TG F-β antibody), Galiximab (anti-CD80 antibody), Gantenerumab nerumab (anti-β-amyloid antibody), gavilimomab (anti-CD147 (basic gin) antibody), gemtuzumab (anti-CD33 antibody), girentuximab ( Anti-carbonic anhydrase 9 antibody), Glembatumumab (also known as CR011, anti-GPN MB antibody), golimumab (also known as Simponi, anti-TNF-α antibody), golimumab (Gomil iximab (anti-CD23C (IgE receptor) antibody), Ibalizumab (anti CD4 antibody), Ibritumomab (anti-CD20 antibody), Igovomab mab) (also known as Indimacis-125, anti-CA-125 antibody), imciromab (also known as Myoscint, an anti-cardiac myosin antibody), infliximab (also known as Remicade, an anti-TNF- α antibody), intetumumab (anti-CD51 antibody), inolimomab mab) (anti-CD25 (IL-2 receptor α chain) antibody), Inotuzumab (anti-C D22 antibody), ipilimumab (anti-CD152 antibody), iratumumab (anti-C D30 (TNFRSF8 antibody), Keliximab (anti-CD4 antibody), label Tuzumab (also known as CEA-Cide, anti-CEA antibody), Lebrikizumab (anti-IL -13 antibody), Lemalesomab (anti-NCA-90 (granulocyte antigen) antibody), Lerdelimumab (anti-TGFβ-2 antibody), Lexatumumab b) (anti-TRAIL-R2 antibody), Libivirumab (anti-hepatitis B surface antigen antibody) body), Lintuzumab (anti-CD33 antibody), Lucatumumab (anti-CD40 antibody), lumiliximab (anti-CD23 (IgE receptor ) antibody), mapatumumab (anti-TRAIL-R1 antibody), maslimomab (anti T cell receptor antibody), Matuzumab (anti-EGFR antibody), Mepolizumab (another Name: Bosatria, anti-IL-5 antibody), Metelimumab (anti-TGFβ-1 antibody) ), Milatuzumab (anti-CD74 antibody), Minretumomab ) (anti-TAG-72 antibody), Mitumomab (also known as BEC-2, anti-ganglionic acid Osidoantibody-GD3), Morolimumab (anti-rhesus factor antibody), Mota Motavizumab (also known as Numax, an anti-RS virus antibody), muromonab b)-CD3 (also known as Orthoclone OKT3, anti-CD3 antibody), nacolomab (anti C242 antibody), Naptumomab (anti-5T4 antibody), Natalizumab (also known as Tysabri, an anti-integrin α4 antibody, Nebacumab (an anti-endotoxin antibody) ), Necitumumab (anti-EGFR antibody), Nerelimomab ( anti-TNF-α antibody), nimotuzumab (also known as Theracim, Theraloc, anti-EGFR antibody), Nofetumomab, ocrelizumab (anti-CD20 antibody), odulimomab ( Alias: Afolimomab, anti-LFA-1 (CD11a) antibody), ofatumumab (also known as Arzerr a, anti-CD20 antibody), olarratumab (anti-PDGF-Rα antibody), Omari Omalizumuba (also known as Xolair, an anti-IgE Fc antibody), oportuzumab (O zumab) (anti-EpCAM antibody), oregovomab (also known as OvaRex, anti-CA- 125 antibody), otelixizumab (anti-CD3 antibody), pagibaximab ( Pagibaximab) (anti-LTA antibody), palivizumab (also known as: Synagis, Abbosynagis, anti-RS virus antibody), panimumab (also known as Vectibix, ABX-EGF, anti-EGFR antibody), Panobacumab (anti-Pseudomonas aeruginosa antibody), Pascolizumab (anti-I L-4 antibody), Pemtumomab (also known as Theragyn, anti-MUC1 antibody), Pel Tuzumab (also known as Omnitarg, 2C4, anti-HER2 / neu antibody), pexelizumab ( lizumab) (anti-C5 antibody), Pintumomab (anti-adenocarcinoma antigen antibody), Pririki Priliximab (anti-CD4 antibody), pritumumab (anti-vimentin antibody) body), PRO140 (anti-CCR5 antibody), racotumomab (also known as 1E1 0, anti-(N-glycolylneuraminic acid (NeuGc, NGNA)-ganglioside (GM 3) Rafivirumab (anti-rabies virus glycoprotein antibody), Ramucirumab (anti-VEGFR2 antibody), ranibizumab (also known as Lucentis, anti-VEGF-A antibody), Raxibacumab (anti-anthrax toxin, protective antigen antibody) ), Regavirumab (anti-CMV glycoprotein B antibody), Reslizumab lizumab (anti-IL-5 antibody), rilotumumab (anti-HGF antibody), Rituki Rituxanmab (also known as MabThera, Rituxanmab, anti-CD20 antibody), Robatumumab ) (anti-IGF-1 receptor antibody), Rontalizumab (anti-IFN-α antibody) , Rovelizumab (also known as LeukArrest, anti-CD11, CD18 antibody), Leukemia Ruplizumab (also known as Antova, anti-CD154 (CD40L) antibody), Satsumo Satumomab (anti-TAG-72 antibody), Sevirumab (anti-CMV antibody) , sibrotuzumab (anti-FAP antibody), sifalimumab (anti-IFN-α antibody) body), Siltuximab (anti-IL-6 antibody), Siplizumab (anti-CD2 antibody) ), (Smart) MI95 (anti-CD33 antibody), solanezumab (anti-β- amyloid antibody), sonepcizumab (anti-sphingosine-1-phosphate antibody) body), Sontuzumab (anti-episialin antibody), Stamulumab ) (anti-myostatin antibody), sulesomab (also known as LeukoScan, (anti-NCA- 90 (granulocyte antigen) antibody), Tacatuzumab (anti-α-fetoprotein antibody), tadocizumab (anti-integrin αIIbβ3 antibody), Mab (anti-IgE antibody), tanezumab (anti-NGF antibody), tapritumomab (t aplitumomab (anti-CD19 antibody), Tefibazumab (also known as Aurexis, Anti-clumping factor A antibody), telimomab, tenatumomab (anti-tenascin-C antibody), teneliximab (anti-CD40 antibody), tepli Zumab (Teplizumab) (anti-CD3 antibody), TGN1412 (anti-CD28 antibody), Ticilim Mab (also known as Tremelimumab, anti-CTLA-4 antibody), Tigatuzumab ( anti-TRAIL-R2 antibody), TNX-650 (anti-IL-13 antibody), tocilizumab (also known as Atlizumab, Actemra, RoActemra, (anti-IL-6 receptor antibody), Toralizumab ab) (anti-CD154 (CD40L) antibody), tositumomab (anti-CD20 antibody), trastuzumab Ibuprofen (also known as Herceptin, an anti-HER2 / neu antibody), Tremelimumab ) (anti-CTLA-4 antibody), Tucotuzumab celmoleukin (anti-EpCAM antibody), tuvirumab (anti-hepatitis B antibody), urtoxazumab Urtoxazumab (anti-Escherichia coli antibody), Ustekinumab kinumab) (also known as Stelara, anti-IL-12, IL-23 antibody), vapaliximab (Vapali ximab) (anti-AOC3 (VAP-1) antibody), vedolizumab, (anti-interferon glin α4β7 antibody), veltuzumab (anti-CD20 antibody), bepalimomab ) (anti-AOC3 (VAP-1) antibody), visilizumab (also known as Nuvion, anti-CD3 antibody) , Vitaxin (anti-angiogenic integrin avb3 antibody), Volociximab ) (anti-integrin α5β1), Votumumab (also known as HumaSPECT, anti-tumor Original CTAA16.88 antibody), Zalutumumab (also known as HuMax-EGFr, (anti-EGFR antibody), Zanolimumab (also known as HuMax-CD4, anti-CD4 antibody), Ziralimumab (anti-C D147 (basic immunoglobulin antibody), zolimomab (anti-CD5 antibody), Etanercept (registered trademark "Enbrel"), Alefacept (registered trademark "Ame") vive), abatacept (registered trademark "Orencia"), rilonacept (Arcal yst), 14F7 [anti-IRP-2 (iron regulatory protein 2) antibody], 14G2a (Nat.Canc er Inst.), anti-ganglioside GD2 antibody for melanoma and solid tumors), J591 (We Anti-PSMA antibodies for treating prostate cancer from Cornell Medical School, 225 .28S [Anti-HMW-MAA (High Molecular Weight Melanoma Associated Antigen) Antibody for Melanoma, Sorin Ra diofarmaci SRL (Milan, Italy)], COL-1 (Colon from Nat. Cancer Inst. Anti-CEACAM3 antibody, CGM1), CYT-356 (registered trademark "On coltad" for prostate cancer), HNK20 (from Ora Vax Inc. for respiratory syncytial virus), Imm uRAIT (from IMMUNOMEDICS for non-Hodgkin's lymphoma), Lym-1 (anti-HLA- DR10 antibody [from Peregrine Pharm for tumors], MAK-195F [from Abbott / Knoll Anti-TNF (tumor necrosis factor; TNFA, TNF-α; TN) for sepsis and toxic shock FSF2) antibody], MEDI-500 [also known as T10B9, from MediImmune Inc. for graft-versus-host disease Anti-CD3 antibody for TRαβ (T cell receptor α / β), RING SCAN [Ne oprobe Corp., an anti-TAG72 (tumor-associated glycoprotein) antibody for breast, colon, and colorectal cancers; Avicidin (anti-EPCAM (epithelial cell adhesion molecule) antibody), anti- TACSTD1 (tumor-associated calcium signal transducer 1) antibody, anti-GA73 3-2 (gastrointestinal tumor-associated protein 2) antibody, anti-EGP-2 (epithelial glycoprotein 2) antibody; Anti-KSA antibody; KS1 / 4 antigen; M4S; tumor antigen 17-1A; colon cancer from NeoRx Corp. , CD326 for ovarian cancer, prostate cancer, and non-Hodgkin's lymphoma; LYMPHOCID E (IMMUNOMEDICS, NJ), Smart ID10 (Protein Design Labs), Oncolym (Tech niclone Inc, CA), Allomune (BioTransplant, CA), anti-VEGF antibody (Genente CE Acide (Immunomedics, NJ); IMC-1C11 (ImClone Sy stems, NJ), and cetuximab (ImClone, NJ).

[0192] Other antibodies as cell binding molecules / ligands include, but are not limited to, the following antigens: Contains: aminopeptidase N (CD13), annexin A1, B7-H3 (CD276 , various cancers), CA125 (ovarian), CA15-3 (carcinoma), CA19-9 (carcinoma), L 6 (carcinoma), Lewis Y (carcinoma), Lewis X (carcinoma), α-fetoprotein (carcinoma), C A242 (colon rectum), placental alkaline phosphatase (carcinoma), prostate-specific antigen (prostate) ), prostatic acid phosphatase (prostate), epidermal growth factor (carcinoma), CD2 (Hodgkin's disease) , NHL lymphoma, multiple myeloma), CD3ε (T-cell lymphoma, lung cancer, breast cancer, gastric cancer, egg CD19 (B-cell malignancies), CD20 (non-Hodgkin's lymphoma), CD22 (leukemia, lymphoma, multiple myeloma, systemic lupus erythematosus), CD30 (Hodgkin's lymphoma), CD33 (leukemia, autoimmune diseases), CD38 (multiple myeloma) myeloma), CD40 (lymphoma, multiple myeloma, leukemia (CLL)), CD51 (metastatic melanoma, sarcoma), CD52 (leukemia), CD56 (small cell lung cancer, ovarian cancer, Merkel cell carcinoma) and liquid tumors, multiple myeloma), CD66e (cancer), CD70 (metastatic renal cell carcinoma and non-homologous tumors) CD74 (multiple myeloma), CD80 (lymphoma), CD98 (cancer) , mucin (carcinoma), CD221 (solid tumor), CD227 (breast cancer, ovarian cancer), CD262 (non-small cell lung cancer and other cancers), CD309 (ovarian cancer), CD326 (solid tumors), CEA CAM3 (colorectal cancer, gastric cancer), CEACAM5 (carcinoembryonic antigen; CEA, CD66e) (breast cancer, colorectal cancer, and lung cancer), DLL4 (Δ-like-4), EGFR (epithelial growth factor) receptors, various cancers), CTLA4 (melanoma), CXCR4 (CD184, hematopoietic tumors, solid tumors) solid tumors), endoglin (CD105, solid tumors), EPCAM (epithelial cell adhesion molecule, bladder bladder, head and neck, colon cancer, NHL prostate cancer, and ovarian cancer), ERBB2 (epidermal growth factor receptor Condition 2: lung cancer, breast cancer, prostate cancer), FCGR1 (autoimmune disease), FOLR (folate receptor , ovarian cancer), GD2 ganglioside (cancer), G-28G (cell surface antigen glycolipid, melanoma) , GD3 idiotype (cancer), heat shock protein (cancer), HER1 (lung cancer, gastric cancer) , HER2 (breast cancer, lung cancer, and ovarian cancer), HLA-DR10 (NHL), HLA-DRB ( NHL, B-cell leukemia), human chorionic gonadotropin (carcinoma), IGF1R (insulin -like growth factor-1 receptor, solid tumors, hematologic cancers), IL-2 receptor (interleukin-2 receptor condition, T-cell leukemia and lymphoma), IL-6R (interleukin 6 receptor, multiple bone myeloma, RA, Castleman's disease, IL6-dependent tumors), integrins (for various cancers) αVβ3, α5β1, α6β4, αIIβ3, α5β5, αVβ5), MAGE-1 (cancer MAGE-2 (carcinoma), MAGE-3 (carcinoma), MAGE-4 (carcinoma), anti-transcriptional Spherin receptor (carcinoma), p97 (melanoma), MS4A1 (transmembrane 4-domain family) Lee A member 1, non-Hodgkin's B-cell lymphoma, leukemia), MUC1 or MUC1-KL H (breast cancer, ovarian cancer, cervical cancer, bronchial and gastrointestinal cancer), MUC16 (CA125) (ovarian cancer) ), CEA (colon), gp100 (melanoma), MART1 (melanoma), MPG (melanoma) , MS4A1 (Transmembrane 4-domain family A member 1, small cell lung cancer, NHL), Creolin, neuro-oncogene product (carcinoma), P21 (carcinoma), anti-(N-glycolylneuraminidase) Paratope of amino acid (breast cancer, melanoma cancer), PLAP-like testicular alkaline phosphatase (ovarian cancer, testicular cancer), PSMA (prostate cancer), PSA (prostate), ROBO4, TAG72 (tumor Tumor-associated glycoprotein 72, leukemia (AML, gastric cancer, colorectal cancer, ovarian cancer), membrane of T cells Transmembrane protein (cancer), Tie (CD202b), TNFRSF10B (tumor necrosis factor receptor receptor superfamily member 10B, cancer), TNFRSF13B (tumor necrosis factor receptor Myeloma superfamily member 13B, multiple myeloma, NHL, other cancers, RA and SLE ), TPBG (trophoblast glycoprotein, renal cell carcinoma), TRAIL-R1 (TNF-related apoptosis) cis-ligand receptor 1, lymphoma, NHL, colorectal cancer, lung cancer), VCAM-1 (CD 106, melanoma), VEGF, VEGF-A, VEGF-2 (CD309) (various cancers) Other tumor-associated antigens recognized by antibodies have already been reported (Gerber, et al. l, mAbs 1:3, 247-253 (2009); Novellino et al, cancer immunol immunother. 54 (3), 187-207 (2005)Franke et al, cancer biother radiopharm. 2000,15,459-76).

[0193] The cell binding agent is preferably a cell-binding agent for binding to tumor cells, virus-infected cells, microorganism-infected cells, parasites, or the like. Live parasite infected cells, autoimmune disease cells, activated tumor cells, bone marrow cells, activated T cells, B cells, More specifically, the cell binding agent can be an antibody to the following: Any agent that can target any cell that expresses either the agonist or one of its receptors / Molecules can be: CD1, CD1a, CD1b, CD1c, CD1d, CD1 e, CD2, CD3, CD3d, CD3e, CD3g, CD4, CD5, CD6, CD7 , CD8, CD8a, CD8b, CD9, CD10, CD11a, CD11b, CD11 c, CD11d, CD12w, CD14, CD15, CD16, CD16a, CD16b , CDw17, CD18, CD19, CD20, CD21, CD22, CD23, CD2 4, CD25, CD26, CD27, CD28, CD29, CD30, CD31, CD3 2, CD32a, CD32b, CD33, CD34, CD35, CD36, CD37, C D38, CD39, CD40, CD41, CD42, CD42a, CD42b, CD42 c, CD42d, CD43, CD44, CD45, CD46, CD47, CD48, CD 49b, CD49c, CD49c, CD49d, CD49f, CD50, CD51, CD 52, CD53, CD54, CD55, CD56, CD57, CD58, CD59, CD 60, CD60a, CD60b, CD60c, CD61, CD62E, CD62L, CD 62P、CD63、CD64、CD65、CD65s、CD66、CD66a、CD66 b、CD66c、CD66d、CD66e、CD66f、CD67、CD68、CD69 、CD70、CD71、CD72、CD73、CD74、CD75、CD75s、CD7 6、CD77、CD78、CD79、CD79a、CD79b、CD80、CD81、C D82、CD83、CD84、CD85、CD85a、CD85b、CD85c、CD8 5d、CD85e、CD85f、CD85g、CD85g、CD85i、CD85j、C D85k、CD85m、CD86、CD87、CD88、CD89、CD90、CD91 、CD92、CD93、CD94、CD95、CD96、CD97、CD98、CD99 、CD100、CD101、CD102、CD103、CD104、CD105、CD1 06、CD107、CD107a、CD107b、CD108、CD109、CD110 、CD111、CD112、CD113、CD114、CD115、CD116、CD1 17、CD118、CD119、CD120、CD120a、CD120b、CD121 、CD121a、CD121b、CD122、CD123、CD123a、CD124、 CD125、CD126、CD127、CD128、CD129、CD130、CD13 1、CD132、CD133、CD134、CD135、CD136、CD137、CD 138、CD139、CD140、CD140a、CD140b、CD141、CD14 2、CD143、CD144、CD145、CDw145、CD146、CD147、C D148、CD149、CD150、CD151、CD152、CD153、CD154 、CD155、CD156、CD156a、CD156b、CD156c、CD156d 、CD157、CD158、CD158a、CD158b1、CD158b2、CD15 8c、CD158d、CD158e1、CD158e2、CD158f2、CD158g 、CD158h、CD158i、CD158j、CD158k、CD159、CD159 a、CD159b、CD159c、CD160、CD161、CD162、CD163、 CD164、CD165、CD166、CD167、CD167a、CD167b、CD 168、CD169、CD170、CD171、CD172、CD172a、CD172 b、CD172g、CD173、CD174、CD175、CD175s、CD176、 CD177、CD178、CD179、CD179a、CD179b、CD180、CD 181、CD182、CD183、CD184、CD185、CD186、CDw186 、CD187、CD188、CD189、CD190、CD191、CD192、CD1 93、CD194、CD195、CD196、CD197、CD198、CD199、C Dw198、CDw199、CD200、CD201、CD202、CD202(a,b )、CD203、CD203c、CD204、CD205、CD206、CD207、C D208、CD209、CD210、CDw210a、CDw210b、CD211、C D212、CD213、CD213a1、CD213a2、CD214、CD215、C D216、CD217、CD218、CD218a、CD218、CD21b9、CD2 20、CD221、CD222、CD223、CD224、CD225、CD226、C D227、CD228、CD229、CD230、CD231、CD232、CD233 、CD234、CD235、CD235a、CD235b、CD236、CD237、C D238、CD239、CD240、CD240ce、CD240d、CD241、CD 242、CD243、CD244、CD245、CD246、CD247、CD248、 CD249、CD250、CD251、CD252、CD253、CD254、CD25 5、CD256、CD257、CD258、CD259、CD260、CD261、CD 262、CD263、CD264、CD265、CD266、CD267、CD268、 CD269、CD270、CD271、CD272、CD273、CD274、CD27 5、CD276、CD277、CD278、CD279、CD281、CD282、CD 283、CD284、CD285、CD286、CD287、CD288、CD289、 CD290、CD291、CD292、CD293、CD294、CD295、CD29 6、CD297、CD298、CD299、CD300、CD300a、CD300b、 CD300c、CD301、CD302、CD303、CD304、CD305、CD3 06、CD307、CD307a、CD307b、CD307c、CD307d、CD3 07e、CD307f、CD308、CD309、CD310、CD311、CD312 、CD313、CD314、CD315、CD316、CD317、CD318、CD3 19、CD320、CD321、CD322、CD323、CD324、CD325、C D326、CD327、CD328、CD329、CD330、CD331、CD332 、CD333、CD334、CD335、CD336、CD337、CD338、CD3 39、CD340、CD341、CD342、CD343、CD344、CD345、C D346, CD347, CD348, CD349, CD350, CD351, CD352 , CD353, CD354, CD355, CD356, CD357, CD358, CD3 59, CD360, CD361, CD362, CD363, CD364, CD365, C D366, CD367, CD368, CD369, CD370, CD371, CD372 , CD373, CD374, CD375, CD376, CD377, CD378, CD3 79, CD381, CD382, CD383, CD384, CD385, CD386, C D387, CD388, CD389, CRIPTO, CRIPTO, CR, CR1, CR GF, CRIPTO, CXCR5, LY64, TDGF1, 4-1BB, APO2, AS LG659, BMPR1B, 4-1BB, 5AC, 5T4 (trophoblastic glycoprotein, TPB G, 5T4, Wnt activation inhibitor 1 or WAIF1), adenocarcinoma antigen, AGS-5, AGS -22M6, activin receptor-like kinase 1, AFP, AKAP-4, ALK, α-integrin Glycine, αvβ6, aminopeptidase N, amyloid β, androgen receptor, and Opoietin 2, angiopoietin 3, annexin A1, anthrax toxin protective antigen, anti Transferrin receptor, AOC3 (VAP-1), B7-H3, Bacillus anthracis, BAFF (B -cell activating factor), BCMA, B-lymphoma cells, bcr-abl, bombesin, BO RIS, C5, C242 antigen, CA125 (carbohydrate antigen 125, MUC16), CA- IX (or CAIX, carbonic anhydrase 9), CALLA, CanAg, canine IL31, carbonic acid Dehydratase IX, cardiac myosin, CCL11 (CC motif chemokine 11), CCR4 (CC chemokine receptor type 4), CCR5, CD3E (epsilon), CEA (carcinoembryonic antigen antigen), CEACAM3, CEACAM5 (carcinoembryonic antigen), CFD (factor D), Ch4 D5, cholecystokinin 2 (CCK2R), CLDN18 (claudin-18), claudin Amplification factor A, cMet, CRIPTO, FCSF1R (colony-stimulating factor 1 receptor) , CSF2 (colony-stimulating factor 2), granulocyte-macrophage colony-stimulating factor (GM-CSF) F), CSP4, CTLA4 (cytotoxic T-lymphocyte-associated protein 4), CTAA1 6.88 Tumor antigen, CXCR4, CXC chemokine receptor type 4, cADP-ribose hydrochloride enzyme, cyclin B1, CYP1B1, cytomegalovirus, cytomegalovirus Glycoprotein B, dabigatran, DLL4 (delta-like ligand 4), DPP4 (dipeptide Dilpeptidase 4), DR5 (death receptor 5), Escherichia coli Shiga toxin type 1, Escherichia coli Shiga Toxin type 2, ED-B, EGFL7 (EGF-like domain-containing protein 7), EGFR, E GFRII, EGFRvIII, endoglin, endothelin B receptor, endotoxin , EpCAM (epithelial cell adhesion molecule), EphA2, episialin, ERBB2 (epithelial growth factor receptor 2), ERBB3, ERG (TMPRSS2ETS fusion gene), Escherichia coli, E TV6-AML, FAP (fibroblast activation protein α), FCGR1, α-fetoprotein Protein, fibrin II, β chain, fibronectin ectodomain B, FOLR (folate receptor) folate receptor α, folate hydrolase, Fos-related antigen 1F, F protein of respiratory syncytial virus Protein, Frizzled receptor, fucosyl GM1, GD2 ganglioside, G-28( Cell surface glycolipid antigen), GD3 idiotype, GloboH, glypican 3, N-glycosyltransferase Cholinneuraminic acid, GM3, GMCSF receptor α chain, growth differentiation factor 8, GP100, G PNMB (transmembrane protein NMB), GUCY2C (guanylate cyclase 2C, guanylate cyclase 2C) Guanylate cyclase C (GC-C), intestinal guanylate cyclase, guanylate cyclase-C receptor, heat-stable enterotoxin receptor (hSTAR), heat shock protein, blood Hepatitis B surface antigen, hepatitis B virus, HER1 (human epidermal growth factor receptor 1) ), HER2, HER2 / neu, HER3(ERBB-3), IgG4, HGF / SF (stem cell growth factor / scatter factor), HHGFR, HIV-1, histone complex, HLA -DA (human leukocyte antigen), HLA-DR10, HLA-DRB, HMWMAA, human chorionic villus Choriogonadotropin, HNGF, human scatter factor receptor kinase, HPV E6 / E7 , Hsp90, hTERT, ICAM-1 (intercellular adhesion molecule 1), idiotype, IG F1R (IGF-1, insulin-like growth factor 1 receptor), IGHE, IFN-γ, Inflammatory IgE, IgE Fc region, IgE, interleukin (IL) -1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-6R, IL-7, I L-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-15, I L-17, IL-17A, IL-18, IL-19, IL-20, IL-21, IL-2 2, IL-23, IL-27, or IL-28), IL-31RA, ILGF2 (IL-23, IL-27, or IL-28), IL-31RA, phospho-like growth factor 2), integrin (α4, α IIIb β3, αvβ3, α4β7, α5 β1, α6β4, α7β7, αIIβ3, α5β5, αvβ5), interferon γ-induced Conductor proteins, ITAGA2, ITGB2 , KIR2D, LCK, Le, legumain, Lewis-Y antigen, LFA-1 (lymphocyte function Related antigen 1, CD11a), LHRH, LINGO-1, lipoteichoic acid, LIV1A, L MP2, LTA, MAD-CT-1, MAD-CT-2, MAGE-1, MAGE-2, MAGE-3, MAGEA1, MAGEA3, MAGE4, ​​MART1, MCP-1, M IF (macrophage migration inhibitory factor or glycosylation inhibitor (GIF)), MS4A1 (Transmembrane 4 domain subfamily A member 1), MSLN (mesothelin), MUC1 (Mucin 1, cell surface associated (MUC1) or polymorphic epithelial mucin (PEM)), MUC1- KLH, MUC16 (CA125), MCP1 (monocyte chemotactic protein 1), Melan A / MART1, ML-IAP, MPG, MS4A1 (transmembrane 4 domain subfamily) -A), MYCN, myelin-associated glycoprotein, myostatin, NA17, NARP-1 , NCA-90 (granulocyte antigen), nectin-4 (ASG-22ME), NGF, neural apo Ptosis-regulatory proteinase 1, NOGO-A, Notch receptor, nucleolin, Ne u Oncogene products, NY-BR-1, NY-ESO-1, OX-40, OxLDL (oxidized low-density lipoprotein (OxLDL) High density lipoprotein), OY-TES1, P21, non-mutant p53, P97, Page 4 , PAP, anti-(N-glycolylneuraminic acid) paratope, PAX3, PAX5, PC SK9, PDCD1 (PD-1, programmed cell death protein 1), PDGF-R α, (platelet-derived growth factor receptor α), PDGFR-β, PDL-1, PLAC1, PL AP-like testicular alkaline phosphatase, platelet-derived growth factor receptor β, sodium phosphate co- Transporter, PMEL17, Polysialic acid, Proteinase 3 (PR1), Prostate cancer, PS( Phosphatidylserine), prostate cancer cells, Pseudomonas aeruginosa, PSMA, PSA, PSCA, rabies Viral glycoproteins, RHD (Rh polypeptide 1 (RhPI)), rhesus factor ( Rhesus factor), RANKL, PhoC, Ras mutants, RG55, RO BO4, respiratory syncytial virus, RON, sarcoma metastasis breakpoint, SART3, sclerostin SLAMF7 (SLAM family member 7), selectin P, SDC1 (synthetic cell death receptor 1), Kan1), sLe(a), somatomedin C, SIP (sphingosine-1-phosphate ), somatostatin, sperm protein 17, SSX2, STEAP1 (sixth membrane of prostate 1) Transepithelial antigen), STEAP2, STn, TAG-22 (tumor-associated glycoprotein 72), Survivin, T cell receptor, T cell transmembrane protein, TEM1 (tumor epithelial marker 1) , TENB2, tenascin C (TN-C), TGF-α, TGF-β (transforming Transforming growth factor β), TGF-β1, TGF-β2 (transforming growth factor β2) , Tie(CD202b), Tie2, TIM-1(CDX-014), TN, TNF, TNF-α, TNFRSF8, TNFRSF10B (tumor necrosis factor receptor superfamily member 10B), TNFRSF13B (tumor necrosis factor receptor superfamily member member 13B), TPBG (trophoblast glycoprotein), TRAIL-R1 (tumor necrosis apoptosis Death-inducing ligand receptor 1), TRAILR2 (death receptor 5 (DR5)), Tumor-specific glycosylation of calcium signal transducer 2 (MUC1), TW EAK receptor, TYRP1 (glycoprotein 75), TRP-2, tyrosinase, VCAM -1, VEGF, VEGF-A, VEGF-2, VEGFR-1, VEGFR2, or Cells expressing Mentin, WT1, XAGE1, or any insulin growth factor receptor; or any epidermal growth factor receptor.

[0194] In another particular embodiment, the cell-binding molecule is a ligand or receptor antigen selected from the following: Folate derivatives (folate receptors, overexpressed in ovarian cancer and other malignancies) may be agonists. binding to proteins) (Low, PS et al 2008, Acc. Chem. Res. 41, 120-129); Urea derivative of tamic acid (binding to prostate-specific membrane antigen, a surface marker of prostate cancer cells) (Hi llier, SM et al, 2009, Cancer Res. 69, 6932-6940); somatostatin (growth hormone) Glycotropin-inhibiting hormone (GHIH), somatotropin-releasing inhibitory factor (SRIF), or somatotropin (also known as tropin-releasing inhibitors) and their derivatives, such as octothreonine Otide (Sandostatin) and Lanreotide (Somatulin) (especially for neuroendocrine tumors, GH producing pituitary adenoma, paraganglionic tumor, nonfunctioning pituitary adenoma, pheochromocytoma) (Ginj, M., et al, 2006, Proc. Natl. Acad. Sci. USA 103, 16436-16441); Somatostatin receptor subtypes (sst1, sst2, sst3, sst4, and sst5)(Reubi JC, Landolt, AM 1984 J. Clin. Endocrinol Metab 59:1148-51; Re ubi JC, Landolt AM 1987 J Clin Endocrinol Metab 65:65-73; Moyse E, et al, J Cli n Endocrinol Metab 61:98-103), gastrointestinal pancreatitis (Reubi JC, et al, 1987 J Clin Endocrinol inol Metab 65:1127-34; Reubi, J. C, et al, 1990 Cancer Res 50:5969-77), brown Chromocyte tumors (Epel-baum J, et al 1995 J Clin Endocrinol Metab 80:1837-44; Reubi JC , et al, 1992 J Clin Endocrinol Metab 74:1082-9), neuroblastoma (Prevost G, 199 6 Neuroendocrinology 63:188-197; Moertel, C. L, et al 1994 Am J Clin Path 102: 752-756), thyroid gland (Reubi, J. C, et al 1991 Lab Invest 64:567-573) Lung cancer (Sagman U, et al, 1990 Cancer 66:2129-2133), myeloma, granuloma, and neuritis Swelling (Reubi JC, et al 1986 J Clin Endocrinol Metab 63:433-8; Reubi JC, et al 1987 Cancer Res 47:5758-64; Fruhwald, M. C, et al 1999 Pediatr Res 45:697-708), Breast cancer (Reubi JC, et al 1990 Int J Cancer 46:416-20; Srkalovic G, et al 1990 J Clin Endocrinol Metab 70:661-669), linpa swelling (Reubi JC, et al 1992, Int J Canc er50:895-900), renal cell carcinoma (Reubi JC, et al 1992, Cancer Res 52:6074-6078), medullary tumors (Reubi JC, et al 1996 Cancer Res 56:1922-31), prostate (Reubi JC, et al 1995, J. Clin. Endocrinol Metab 80 :2806-14; et al 1989, Prostate 14:191-208; Halmos G, et al J. Clin. Endo-crinol Metab 85:2564-71), ovary (Halmos, G, et a l, 2000 J Clin Endocrinol Metab 85:3509-12; Reubi JC, et al 1991 Am J Pathol 13 8:1267-72), stomach (Reubi JC, et al 1999, Int J Cancer 81: 376-86; Miller, G. V. 1992 Br J Cancer 66:391-95), hepatocytes (Kouroumalis E, et al 1998 Gut 42:442-7; Reubi JC, et al. 1999 Gut 45:66-774) and nasopharyngeal carcinoma (Loh K. S, et al. 2002 Virch ows Arch 441:444-8; specific for carbonic anhydrase IX (a marker of hypoxia and renal cell carcinoma) Certain aromatic sulfonamides (Neri, D., et al., Nat. Rev. Drug Discov. 2011, 10, 767-777); Pituitary adenylate cyclase activation for pheochromocytoma and paraganglioma tumors Peptide (PACAP) (PAC1); Vasoactive intestinal peptide (VIP / PACAP) and its receptor subtypes (VPAC1, VCAP2); α-melanocyte-stimulating hormone ( α-MSH receptor; cholecystokinin (CCK) / gastrin receptor and its receptor subtypes types (CCK1 (formerly CCK-A) and CCK2; bombesin (Pyr-Gln-Arg-Leu-Gl y-Asn-Gln-Trp-Ala-Val-Gly-H is-Leu-Met-NH2) / gastrin-releasing peptide (GRP) ( BB1, GRP receptor subtype (BB2), BB3, and BB4) (Ohlsson, B., et al, 1999, Scand. J. Gastroenterology 34 (12): 1224-9; Weber, HC, 2009, Cur. Opin. Endocri. Diab. Obesity 16(1): 66-71, Gonzalez N, et al, 2008, Cur. Opin. E Neurotensin receptors and their receptor subtypes Substance P receptors and their receptor subtypes (NTR1, NTR2, NTR3); NK1 receptors in glial tumors, etc.), Hennig IM et al, 1995 Int. J. Cancer 61, 786-792 ); Neuropeptide Y (NPY) receptor and its receptor subtypes (Y1-Y6); R GD (Arg-Gly-Asp), NGR (Asn-Gly-Arg), dimeric and multimeric cyclic RGD peptides ( For example, cRGDfV)(Laakkonen P, Vuorinen K. 2010, Integr Biol (Camb). 2(7-8 ): 326-337; Chen K, Chen X. 2011, Theranostics. 1:189-200; Garanger E, et al, An ti-Cancer Agents Med Chem. 7 (5): 552-558; Kerr, JS et al, Anticancer Research h, 19(2A), 959-968; Thumshirn, G, et al, 2003 Chem. Eur. J. 9, 2717- 2725), and TAASGVRSMH and LTLRWVGLMS (chondroitin sulfate proteoglycans) 31) that binds to the cell surface-expressed nucleolin receptor (NG2 receptor) and F3 peptide ( Amino acid peptide) (Zitzmann, S., 2002 Cancer Res., 62, 18, pp. 5139-5143, Temmi nga, K., 2005, Drug Resistance Updates, 8, 381-402; P. Laakkonen and K. Vuorine n, 2010 Integrative Biol, 2(7-8), 326-337; MA Burg, 1999 Cancer Res., 59(12), 2869-2874; K. Porkka, et al 2002, Proc. Nat. Acad. Sci. USA 99(11), 7444-9) Homing peptides, including cell-penetrating peptides (CPPs) (Nakase I, et al, 2012, J. Control Release. 159(2),181-188); Peptide hormones, e.g., testosterone production as well as by targeting follicle-stimulating hormone (FSH) and luteinizing hormone (LH). Luteinizing hormone-releasing hormone (LHRH) agonists and antagonists, and gonadotropin-releasing hormone (GnRH) agonists, such as buserelin (Pyr-Hi s-Trp-Ser-Tyr-D-Ser(OtBu)-Leu-Arg-Pro-NHEt), gonadorelin (Pyr-His-Trp-Ser-Tyr- Gly-Leu-Arg-Pro-Gly-NH2), goserelin (Pyr-His-Trp-Ser-Tyr-D-Ser(OtBu)-Leu-Arg-P ro-AzGly-NH2), histrelin (Pyr-His-Trp-Ser-Tyr-D-His(N-benzyl)-Leu-Arg-Pro-NH Et), leuprolide (Pyr-His-Trp-Ser-Tyr-D-Leu-Leu-Arg-Pro-NHEt), nafarelin ( Pyr-His-Trp-Ser-Tyr-2Nal-Leu-Arg-Pro-Gly-NH2), triptorelin (Pyr-His-Trp-Ser- Tyr-D-Trp-Leu-Arg-Pro-Gly-NH2), nafarelin, deslorelin, abarelix (Ac- D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-(N-Me)Tyr-D-Asn-Leu-isopropylLys-Pro -DAla-NH2), Cetrorelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-Ty rD-Cit-Leu-Arg-Pro-D-Ala-NH2), degarelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3- pyridyl)Ala-Ser-4-aminoPhe(L-hydroorotyl)-D-4-aminoPhe(carba-moyl)-Leu-isopropyl Lys-Pro-D-Ala-NH2), and ganirelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl) Ala-Ser-Tyr-D-(N9, N10-diethyl)-homoArg-Leu-(N9, N10-diethyl)-homoArg-Pro-D-Ala- NH2)(Thundimadathil, J., J. Amino Acids, 2012, 967347, doi:10.1155 / 2012 / 96734 7; Boccon-Gibod, L.; et al, 2011, Therapeutic Advances in Urology 3 (3): 127-140 ; Debruyne, F., 2006, Future Oncology, 2(6), 677-696; Schally A. V; Nagy, A. 199 9 Eur J Endocrinol 141:1-14; Koppan M, et al 1999 Prostate 38:151-158); For example, small molecules (imiquimod, guanidine and adenosine analogues) and lipopolysaccharide ( LPS), nucleic acids (CpG DNA, poly I;C) and lipopeptides (Pam3CSK4) (Kasturi, SP, et al, 2011, Nature 470, 543-547; Lane, T., 2001, JR Soc. Med. 94, 316; Hotz, C., and Bourquin, C., 2012, Oncoimmunology 1, 227-228; Dudek , AZ, et al, 2007, Clin. Cancer Res. 13, 7119-7125) Toll-like receptors (TLRs), C-type lectins, and Nodl recognize molecules down to the macromolecular scale ike receptors (NLRs) (Fukata, M., et al, 2009, Semin. Immunol. 21, 242-253; Maisonneuve, C., et al, 2014, Proc. Natl. Acad. Sci. USA 111, 1-6; Botos, I., et al, 2011, Structure 19, 447-459; Means, TK, et al, 2000, Life Sci. 68, 241-258) and other pattern recognition receptors (PRRs), primarily through their effects on osteoclasts and the kidney. Calcitonin receptor, a 32-amino acid neuropeptide involved in the regulation of calcium levels. body (Zaidi M, et al, 1990 Crit Rev Clin Lab Sci 28, 109-174; Gorn, AH, et al. 1995 J Clin Invest 95:2680-91); Integrin receptors generally play an important role in angiogenesis The body and its receptor subtypes (α V β1, α V β3, α V β5, α V β6, α6β4, α7β1, α L β2 , α IIb β3, etc.) are expressed on the surface of various cells, particularly osteoclasts, endothelial cells, and tumor cells. (Ruoslahti, E. et al, 1994 Cell 77, 477-8; Albelda, SM et al, 1990 Cancer Res. , 50, 6757-64). Short peptides such as GRGDSPK and cyclic (RGDfV) (L1) Cyclic RGD pentapeptide and its derivatives [cyclo(-N(Me)R-GDfV), cyclo (R-Sar-DfV), cyclo(RG-N(Me)D-fV), cyclo(RGD-N( cyclo(RGDf-N(Me)V-) (cilengitide)] is an integrin showed high binding affinity to the steroid receptor (Dechantsreiter, MA et al., 1999 J. Med. Chem. 42 , 3033-40, Goodman SL et al, 2002 J. Med.Chem.45, 1045-51).

[0195] Cell-binding molecules / ligands or cell receptor agonists include Ig-based and non-Ig-based The Ig-based scaffold can be, but is not limited to, a protein scaffold molecule. However, nanobodies (VHH (camelid Ig) derivatives) (Muyldermans S., 2013 Annu Re v Biochem. 82, 775-797; domain antibodies (dAb, derivatives of VH or VL domains) ( Holt, L. J, et al, 2003, Trends Biotechnol. 21, 484-490); Gauger (BiTE, bispecific dimer) (Baeuerle, P. A, et al, 2009, Curr. Opin. M ol. Ther. 11, 22-30); dual affinity retargeting (DART, bispecific dimer) (Moore P A. P, et al. 2011, Blood 117(17), 4542-4551; tetravalent tandem antibody (TandAb , bispecific dimer) (Cochlovius, B, et al. 2000, Cancer Res. 60(16):4336-4341) Non-Ig scaffolds can be selected from, but are not limited to, anticalins ( Lipocalin derivatives) (Skerra A. 2008, FEBS J., 275(11): 2677-2683; Beste G, et al, 1999 Proc. Nat. Acad. USA. 96(5):1898-1903; Skerra, A. 2000 Biochim Biophys Acta, 1482(1-2):337-350; Skerra, A. 2007, Curr Opin Biotechnol. 18(4):295-304; Skerra, A. 2008, FEBS J. 275(11):2677-2683; Adnectins (FN3 (Finally bronectin)) (Koide, A, et al, 1998 J. Mol. Biol., 284(4):1141-1151; Batori V, 2002, Protein Eng. 15(12): 1015-1020; Tolcher, A. W, 2011, Clin. Cancer Re s. 17(2):363-371; Hackel, B. J, 2010, Protein Eng. Des. Sel. 23(4):211-219); Developed ankyrin repeat proteins (DARPins) (ankyrin repeat (AR) ) Protein derivatives) (Boersma, YL, et al, 2011 Curr Opin Biotechnol. 22(6): 8 49-857), for example, DARPinC9, DARPinEc4, and DARPinE69_ LZ3_E01(Winkler J, et al, 2009 Mol Cancer Ther. 8(9), 2674-2683; Patrici a MK. M., et al, Clin Cancer Res. 2011;17(1):100-110; Boersma Y. L, et al, 2011 J. Biol. Chem. 286(48),41273-41285; Avimer (Domain A / low High density lipoprotein (LDL) receptor) (Boersma Y. L, 2011 J. Biol. Chem. 286(48): 41273-41285; Silverman J, et al, 2005 Nat. Biotechnol., 23(12):1556-1561) You can choose.

[0196] Examples of small molecule structures of cell binding molecules / ligands or cell receptor agonists of the present patent application are: The structures are as follows: LB01 (folic acid), LB02 (PMSA ligand) , LB03 (PMSA ligand), LB04 (PMSA ligand), LB05 (Somatos statin), LB06 (somatostatin), LB07 (octreotide, somatostatin analogs) analogue), LB08 (lammatide, somatostatin analogue), LB09 (vapreotide (saprost) member), somatostatin analogue), LB10 (CAIX ligand), LB11 (CAI X ligand), LB12 (gastrin-releasing peptide receptor (GRPr, MBA), LB 13 (luteinizing hormone-releasing hormone (LH-RH) ligand and GnRH), LB14 ( Luteinizing hormone-releasing hormone (LH-RH) and GnRH ligand), LB15 (GnR H antagonist, Abarelix), LB16 (cobalamin, vitamin B12 analogue ), LB17 (cobalamin, vitamin B12 analogue), LB18 (α v β3 integrin receptor, cyclic RGD pentapeptide), LB19 (hetero-bivalent peptide of VEGF receptor) ligand), LB20 (neuromedin B), LB21 (G-protein-coupled receptor LB22 (Toll-like receptor 2 (TLR2)), LB23 (androgen receptor for the αv integrin receptor), LB24 (cilengitide / cyclo(- RGDfV-), LB23 (flurocortisone), LB25 (rifabutin analogue), LB26 (rifabutin analogue), LB27 (rifabutin analogue), LB28 (fludoxetine analogue) LB29 (dexamethasone), LB30 (fluticasone propionate) , LB31 (beclomethasone dipropionate), LB32 (triamcinolone acetonide) ), LB33 (Prednisone), LB34 (Prednisolone), LB35 (Methylprednisolone) Nisolone), LB36 (betamethasone), LB37 (irinotecan analogue), LB38 ( Crizotinib analogue), LB39 (bortezomib analogue), LB40 (carfilzomib analogue) LB41 (carfilzomib analogue), LB42 (leuprolide analogue), LB43 (triptorelin analogue), LB44 (clindamycin), LB45 (liraglutide) LB46 (semaglutide analogue), LB47 (retapamulin analogue), LB48 (indibulin analogue), LB49 (vinblastine analogue), LB50 (liquid Sisenatide analogue), LB51 (osimertinib analogue), LB52 (nucleoside analogue) LB53 (erlotinib analog), and LB54 (lapatinib analog):

[0197] [ka] TIFF0007637431000087.tif209156TIFF0007637431000088.tif207136TIFF0007637431000089.tif235153TIFF0007637431000090.tif201152 TIFF0007637431000091.tif221157TIFF0007637431000092.tif214165TIFF0007637431000093.tif222158TIFF0007637431000094.tif189151

[0198] During the ceremony, [ka] is the site for linking to the side chain linker of the present invention;

[0199] X4 and Y1 are independently O, NH, NHNH, NR1, S, C(O)O, C(O)N H, OC(O)NH, OC(O)O, NHC(O)NH, NHC(O)S, OC(O)N (R1), N(R1)C(O)N(R1), CH2, C(O)NHNHC(O), and C (O) is NR1;

[0200] X1 is H, CH2, OH, O, C(O), C(O)NH, C(O)N(R1), R1, NHR1, NR1, C(O)R1, or C(O)O;

[0201] X5 is H, CH3, F, or Cl;

[0202] M1 and M2 are independently H, Na, K, Ca, Mg, NH4, N(R1R2R3R4 ) wherein R1, R2, R3, and R4 are as defined in formula (I).

[0203] Applications of conjugates

[0204] In certain embodiments, the cell-binding ligand-drug conjugates of the present invention via side chain linkers comprise: It is used in targeted cancer therapy. Targeted cancers include, but are not limited to: Adrenal cortical carcinoma, anal cancer, bladder cancer, brain tumors (adults, brainstem glioma, children, cerebellar stellate cell cystoma, brain astrocytoma, ependymoma, medulloblastoma, supratentorial primitive neuroectodermal and pineal tumor, optic tract and hypothalamic glioma), breast cancer, carcinoid tumor, gastrointestinal, carcinoma of unknown primary, cervical cancer tumors, colorectal carcinoma, endometrial cancer, esophageal cancer, extrahepatic bile duct cancer, Ewing family tumors (PNET), extracranial malignant germ cell tumor, eye cancer, intraocular melanoma, gallbladder cancer, gastric cancer (stomach), Germ cell tumor, extragonadal, gestational trophoblastic tumor, head and neck cancer, hypopharyngeal cancer, pancreatic islet cell carcinoma, kidney Cancer (renal cell carcinoma), laryngeal carcinoma, leukemia (acute lymphoblastic, acute myeloid, chronic lymphocytic) cancer of the lip and oral cavity, liver cancer, lung cancer (non-small cell, small cell), Cell, lymphoma (AIDS-related, central nervous system, cutaneous T-cell, Hodgkin's disease, non-Hodgkin's disease, malignant mesothelioma, melanoma, Merkel cell carcinoma, metastatic squamous neck cancer of unknown primary origin, multiple myeloma, and other plasma cell neoplasms, mycosis fungoides, myelodysplastic syndromes, myeloproliferative syndromes, nasopharyngeal carcinoma , neuroblastoma, oral cancer, pharyngeal cancer, osteosarcoma, ovarian cancer (epithelial, germ cell tumors, low malignant potential tumors), pancreatic cancer (exocrine gland, islet cell cancer), paranasal sinus and nasal cavity cancer, adenocarcinoma, penile cancer, pheochromocytoma, pituitary cancer, plasmacytoma, prostate cancer, striated muscle rectal cancer, renal cell carcinoma (kidney cancer), renal pelvis and ureter (transitional cell), salivary gland cancer, Sezary syndrome, skin cancer, skin cancer (cutaneous T-cell lymphoma, Kaposi's sarcoma, melanoma), small intestine cancer , soft tissue sarcoma, stomach cancer, testicular cancer, thymoma (malignant), thyroid cancer, urethral cancer, uterine cancer (sarcoma), unusual cancers of children, including vaginal cancer, vulvar cancer, and Wilms' tumor.

[0205] In another specific embodiment, the cell-binding drug conjugate of the present invention, by its components and method, It can be used for the treatment or prevention of autoimmune diseases. The autoimmune diseases are not limited to these. However, autoimmune gastric achlorhydria, chronic active hepatitis, acute disseminated encephalomyelitis, acute hemorrhagic leukoencephalopathy, inflammation, Addison's disease, agammaglobulinemia, alopecia areata, amyotrophic lateral sclerosis, ankylosing spine Spondylitis, anti-GMB / TBM nephritis, antiphospholipid syndrome, antisynthetase syndrome, arthritis , atopic allergy, atopic dermatitis, autoimmune aplastic anemia, autoimmune heart disease Myopathy, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inner ear disease, autoimmune lymphoma globulin proliferative syndrome, autoimmune peripheral nervous system disease, autoimmune pancreatitis, multiple autoimmune endocrine disorders Type I, II, III, autoimmune progesterone dermatitis, autoimmune thrombocytopenic purpura Disease, autoimmune uveitis, Barlow's disease / Barlow concentric sclerosis, Behçet's disease, Be rger disease, Bickerstaff encephalitis, Blau syndrome, bullous pemphigoid, Cass Lemann's disease, Chagas disease, chronic fatigue and immune dysfunction syndrome, chronic inflammatory demyelinating polyneuropathy Harm, chronic recurrent multifocal osteomyelitis, chronic Lyme disease, chronic obstructive pulmonary disease, allergic granuloma vasculitis, cicatricial pemphigoid, celiac disease, Cogan's syndrome, cold agglutinin disease, complement component C 2 deficiency, cephalic arteritis, CREST syndrome, Crohn's disease (idiopathic inflammatory bowel disease), Cussinus disease syndrome, cutaneous leukocytoclastic vasculitis, malignant atrophic papulopathy, painful steatosis, dermatitis herpetiformis, cutaneous Dermatomyositis, type 1 diabetes, diffuse cutaneous scleroderma, myocardial infarction, discoid lupus erythematosus, eczema, uterine Endometriosis, enthesitis-associated arthritis, eosinophilic fasciitis, epidermolysis bullosa acquisita, erythema nodosum, idiopathic Mixed cryoglobulinemia, Evans syndrome, fibrodysplasia ossificans progressiva, fibromyalgia , fibromyositis, fibrosing alveolitis, gastritis, gastrointestinal pemphigoid, giant cell arteritis, glomerulonephritis, Goodpasture's syndrome, Graves' disease, Guillain-Barré syndrome, Hashimoto's encephalopathy, Hashimoto's thyroiditis , hemolytic anemia, allergic purpura, herpes gestationis, hidradenitis suppurativa, Hughes syndrome (anti- phospholipid syndrome, hypogammaglobulinemia, idiopathic inflammatory demyelinating disease, idiopathic pulmonary fibrosis , idiopathic thrombocytopenic purpura (autoimmune thrombocytopenic purpura), IgA nephropathy (Berg er's disease), inclusion body myositis, inflammatory demyelinating polyneuropathy, interstitial cystitis, irritable bowel syndrome, Juvenile idiopathic arthritis, juvenile rheumatoid arthritis, mucocutaneous lymph node syndrome, Lambert-Y Myasthenic syndrome, leukocytoclastic vasculitis, lichen planus, lichen sclerosus, linear IgA disease (L AD), Lou Gehrig's disease (amyotrophic lateral sclerosis), lupus-like hepatitis, lupus erythematosus, Bleu syndrome, Meniere's disease, microscopic polyangiitis, Miller-Fisher syndrome, mixed connective tissue Osteoarthritis, scleroderma, Mucha-Jakob disease, Muckle-Wells syndrome, multiple myeloma, multiple myeloma Sclerosis, myasthenia gravis, myositis, narcolepsy, neuromyelitis optica (Devic's disease), neuromuscular , Ocular cicatricial pemphigoid, Opsoclonus-myoclonus syndrome, Ord's thyroiditis, Regression Rheumatism, Panda syndrome (childhood autoimmune neuropsychiatric disorder complicated by streptococcal infection), small tumors Cerebral degeneration, paroxysmal nocturnal hemoglobinuria, Parry-Romberg syndrome, Parsonage-George Pemphigus syndrome, pars planus inflammation, pemphigus vulgaris, pernicious anemia, perivenous encephalomyelitis, POE MS syndrome, polyarteritis nodosa, polymyalgia rheumatica, polymyositis, primary biliary cirrhosis , primary sclerosing cholangitis, progressive inflammatory neuropathy, psoriasis, psoriatic arthritis, pyoderma gangrenosum, pure Aplastic anemia, Rasmussen's encephalitis, Raynaud's disease, relapsing polychondritis, Reiter's disease syndrome, restless legs syndrome, retroperitoneal fibrosis, rheumatoid arthritis, rheumatic fever, sarcoidosis Schizophrenia, Schmidt syndrome, Schnitzler syndrome, scleritis, scleroderma, Schoenfeld syndrome Ren's syndrome, spondyloarthropathy, sticky blood syndrome, Still's disease, stiff-man syndrome, subcutaneous Acute bacterial endocarditis, Susac syndrome, acute febrile neutrophilic dermatosis, Sydenham chorea, sympathetic Ophthalmitis, Takayasu's arteritis, temporal arteritis (giant cell arteritis), Tolosa-Hunt syndrome, transverse spinal cord inflammation, ulcerative colitis (idiopathic inflammatory bowel disease), undifferentiated connective tissue disease, undifferentiated spondyloarthropathy, blood vessels vitiligo, Wegener's granulomatosis, Wilson's syndrome, Westcott-Aldrich syndrome Includes groups.

[0206] In another specific embodiment, the side chain conjugates of the invention for the treatment or prevention of autoimmune diseases are Linking molecules used for conjugation via ELISA include anti-elastin antibodies; Abys anti-epithelial cells; Antibodies; anti-basement membrane type IV collagen protein antibodies; antinuclear antibodies; anti-double-stranded DNA antibodies, anti Single-stranded DNA antibody, anticardiolipin antibody IgM, IgG; anti-celiac antibody; antiphospholipid antibody Anti-mitochondrial antibodies IgK, IgG; anti-SM antibodies; anti-mitochondrial antibodies; thyroid antibodies; microsomes Antibodies, T-cell antibodies; thyroglobulin antibodies, anti-scleroderma-70 antibodies (AntiSCL-70) Anti-Jo antibody (Anti-Jo), anti-U1RNP antibody (Anti-U1RNP); anti-L a / SSB antibody; anti-SSA antibody; anti-SSB antibody; anti-parietal cell antibody; anti-histone antibody; anti-RN antibody P antibody; C-ANCA; P-ANCA; anti-centromere antibody; anti-fibrin antibody, anti-GB antibody M antibody, anti-ganglioside antibody; anti-desmosomal glycoprotein 3 core antibody (anti-D esmogein3); anti-p62 antibody; anti-sp100 antibody; anti-mitochondrial (M2) antibody Rheumatoid factor antibody; anti-MCV antibody; anti-topoisomerase antibody; anti-neutrophil cytoplasmic antibody (cA These include, but are not limited to, NCA antibodies.

[0207] In some preferred embodiments, the binding molecules used in the conjugates of the invention are By binding to receptors or receptor complexes expressed by disease-associated activated lymphocytes. The receptor or receptor complex can be, for example, a member of the immunoglobulin gene superfamily. Members of the CD11 receptor (e.g., CD2, CD3, CD4, CD8, CD19, CD20, CD 22, CD28, CD30, CD33, CD37, CD38, CD56, CD70, CD 79, CD90, CD125, CD147, CD152 / CTLA-4, PD-1, or ICOS), TNF receptor superfamily (e.g., CD27, CD40, CD95 / Fas, CD134 / OX40, CD137 / 4-1BB, INF-R1, TNFR- 2, RANK, TACI, BCMA, osteoprotegerin, Apo2 / TRAIL-R 1, TRAIL-R2, TRAIL-R3, TRAIL-R4, and APO-3), Tegrin, cytokine receptors, chemokine receptors, major histocompatibility proteins, lectins Examples of such proteins include complement regulatory proteins (type C, type S, or type I).

[0208] In another specific embodiment, the antibody has immunospecificity for a viral or microbial antigen. Useful cell-binding ligands are humanized or human monoclonal antibodies. The term "viral antigen" as used herein includes any viral peptide capable of eliciting an immune response. polypeptide proteins (e.g., HIV gp120, HIV nef, RSV F glycoproteins) Protein, influenza virus neuraminidase, influenza virus hemagglutination proteins, HTLV-Tac, herpes simplex virus glycoproteins (e.g., gB, gC, gD, and These include, but are not limited to, hepatitis B surface antigen (HSA), hepatitis C antigen (HSA), and hepatitis B surface antigen (HSA). The term "microbial antigen" as used herein includes any microbial peptide, polypeptide, or polypeptide that can elicit an immune response. peptides, proteins, sugars, polysaccharides, or lipid molecules (e.g., bacteria, fungi, pathogenic organisms) Animal and yeast polypeptides (e.g., LPS and capsular polysaccharide 5 / 8) are included, but these Examples of antibodies useful for treating viral or microbial infections include, but are not limited to, palivizumab (humanized anti-respiratory syncytial virus monoclonal antibody used to treat RVS infection), P RO542 (CD4 fusion antibody used to treat HIV infection), Ostavir (hepatitis B virus) (human antibody used to treat cytomegalovirus), PROTVIR (human antibody used to treat cytomegalovirus) These include, but are not limited to, humanized antibodies (IgG1 antibodies), and anti-LPS antibodies.

[0209] The cell-binding molecule-side chain linked drug conjugate of the present invention can be used to treat infectious diseases. These infections include Acinetobacter infection, actinomycosis, and African sleeping sickness. (African trypanosomiasis), AIDS (acquired immune deficiency syndrome), amebiasis, Plasma, anthrax, bacterial tuberculosis infection, Argentine hemorrhagic fever, ascariasis, aspergillosis, aspergillosis Torovirus infection, babesiosis, Bacillus cereus infection, bacterial pneumonia, bacterial vaginosis, bacterium roides infection, Balantidiosis, Bailey's roundworm infection, BK virus infection, black sand Hair, Blastocys hominis infection, Blastomyces, Bolivian hemorrhagic fever, Borrelia infection, Botulism (and infant botulism), Brazilian hemorrhagic fever, brucellosis, Burkholderia Buruli ulcer, Calicivirus (Norovirus, Sapovirus), Pyrobacter infection, Candida infection (candidiasis, thrush), cat scratch disease , cellulitis, Chagas disease (American trypanosomiasis), chancroid, chickenpox, lichen planus, pneumonia Clostridium difficile infection, cholera, pigmented fungus, liver fluke disease, Clostridium difficile infection, Csidiidomycosis, Colorado tick fever, colds (acute viral nasopharyngitis, acute rhinitis), Itzfeldt-Jakob disease, Crimean-Congo hemorrhagic fever, Cryptococcus, Cryptosporidium Diarrhea, cutaneous larval migration, Cyclospora infection, cysticercosis, cytomegalovirus infection, Dengue fever Guinea fever, diamoebiasis, diphtheria, diphyllobothriasis, dracunculiasis, Ebola hemorrhagic fever, hydatid disease, Ehrlichiosis, pinworms (pinworm infection), enterococcal infection, enterovirus infection, rash Fus, erythema infectiosum (fifth disease), acute rash in children, hypertrophic paragonimiasis, trematodia, fatal familial insomnia disease, filariasis, food poisoning caused by Clostridium perfringens, non-parasitic amoeba infections , Fusobacterium infection, gas gangrene (clostridial myonecrosis), diotrichosis, Lustmann-Straussler-Scheinker syndrome, Giardiasis, Glanders, Gnathostomiasis , gonorrhea, granuloma groin (donovanosis), group A streptococcal infection, group B streptococcal infection, Influenza, hand, foot and mouth disease (HFMD), Hantavirus pulmonary syndrome, Helicobacter pylori infection, hemolytic uremic syndrome, hemorrhagic fever with renal syndrome, hepatitis A, hepatitis B, hepatitis C, D Hepatitis, Hepatitis E, Herpes simplex, Histoplasmosis, Hookworm infection, Human Balkan virus Infection, Human Ehrlichiosis Evans, Human Granulocytic Anaplasmosis, Human Metapneumocystis human papillomavirus infection, human monocytic amyloidosis, human papillomavirus infection, human parainfluenza virus infection Vancomycin virus infection, microtaeniasis, influenza, isosporosis, Kawasaki disease, mononuclear cells disease, Kim's disease, kuru, Lassa fever, Legionnaires' disease, Legionnaires' disease Anticoagulant fever), leishmaniasis, leprosy, leptospirosis, listeriosis, Lymphocytic choriomeningitis, Lymphocytic borrelia, Lymphatic filariasis (elephantiasis), Laria, Marburg hemorrhagic fever, measles, melioidosis (Whitmore's disease), meningitis, meningococcal disease Diseases: metagonism, microsporidiosis, molluscum contagiosum, mumps, typhus (endemic) typhus), mycoplasma pneumonia, mycetoma, myiasis, neonatal conjunctivitis (ophthalmia neonatorum), Itzfeldt-Jakob disease (vCJD, nvCJD), nocardiosis, onchocerciasis (loss of consciousness) Active filariasis), paracoccidioidomycosis (South American blastomycosis), paragonimiasis, Turela disease, head lice (head lice), body lice (body lice), pubic lice Cough (pubic lice, Crambs ice), pelvic inflammatory disease, whooping cough ough), plague, pneumococcal infection, Pneumocystis carinii pneumonia, pneumonia, polio, Prevotella infection, P AME, progressive multifocal leukoencephalopathy, psittacosis, Q fever, rabies, rat bite fever, respiratory syncytial syndrome Somatic virus infection, rhinovirus infection, rickettsial infection, rickettsia, Rift Valley fever , Rocky Mountain spotted fever, rotavirus infection, rubella, salmonellosis, SARS (severe acute respiratory syndrome), Haemorrhagic syndrome), scabies, schistosomiasis, septicemia, diarrhea (dysentery), shingles (Herpes zoster) oster), smallpox, sporotrichum, staphylococcal food poisoning, staphylococcal infection, nematodes, plum Poison, Taeniasis, Tetanus (Trismus), Barber's itch, White Hands tinea versicolor, pityriasis nigricans, tinea pedis, tinea unguium, tinea versicolor, toxocariasis (ocular larva migrans), visceral larva migrans, toxoplasmosis, trichinosis, trichomoniasis, cryptobiosis whipworm infection, pulmonary tuberculosis, tularemia, urea-degrading urea Mycoplasma infection, Venezuelan Encephalitis, Venezuelan hemorrhagic fever, viral pneumonia, West Nile fever, rhizobacterial disease, pseudotuberculosis Fungal infections include, but are not limited to, yersiniosis, yellow fever, and zygomycosis.

[0210] More preferably, the cell-binding molecule is an antibody against a pathogenic strain described herein, The pathogenic strains include Acinetobacter baumannii, Actiomyces israelii, and Acinetobacter pneumoniae. Actinomyces odontolyticus s), Propionibacterium propionicus, Trypanosoma brucei, HIV (human immunodeficiency virus), Entamoeba histolytica, Anaplasma spp., Bacillus anthracis, Methylshemolites Arcanobacterium haemolyticum, Junin virus Aspergillus, Ascaris, Astroviridae, Babesia, Bacillus cereus, Malaria Tipul bacteria, Bacteroides spp., Colonic pouch ciliates, Bailey roundworms, Nematode spp., BK Virus, Piedraiahortae, Blastocyst Diphtheria hominis, Dermatitis gracilis, Makupo virus, Borrelia spp., Clostridium botulinum, Sabia , Brucella spp., typically Burkholderia cepacia and other Burkholderia species, Mycobacterium Caliciviridae family, Campylobacter, commonly known as Caliciviridae Candida albicans and other Candida species, Bartonella henselae ella henselae, group A streptococci and staphylococci, Trypanosoma cruzi , Haemophilus chancroid, Varicella-zoster virus (VZV), Chlamydia trachomatis, Chlamydia A. pneumoniae, Vibrio cholerae, Fonseca pedrosoi, Clonorchiasis, Clostridium difficile, Coccidioides immitis, Coccidioides posadaci, Corola Tick ​​fever virus, rhinovirus, coronavirus, Creutzfeldt-Jakob disease Lyon, Crimean-Congo hemorrhagic fever virus, Cryptococcus neoformans, Crimean-Congo hemorrhagic fever virus ... Ptosporidium, cat hookworm, co-parasites, Cyclospora, Taenia solium, Cytomegalovirus dengue viruses (DEN-1, DEN-2, DEN-3 and DEN-4) - flavivirus Viruses, Dimorphonucleate amoeba, Corynebacterium diphtheriae, Diphyllobothria, Dracunculia spp., Guinea worm (D racunculusmedinensis), Ebola virus, Echinococcus spp., - Rickettsia spp., pinworms, Enterococcus spp., Enterovirus spp., Rickettsia typhi , Parvovirus B19, Human Herpesvirus 6, Human Herpesvirus 7, Hypertrophy Flukes, Liver Fluke and Liver Fluke Giant, FFI Prion, Filariae, Clostridium perfringens, Fuso Bacterium, Clostridium perfringens, other Clostridium species, Geotrichum candidum, G SS prion, Giardia lamblia, Burkholderia Burkholderia mallei, Gnathostoma nematode, Gnathostoma strigiforme, Neisseria gonorrhoeae, Bordetella granulomatosis, Streptococcus pyogenes, Streptococcus pyogenes agalactiae, Haemophilus influenzae, enteric viruses, most Coxsackieviruses A Enterovirus type 71, Sin Nombre virus, Helicobacter pylori, Escherichia coli O 158:H7, Bunyaviridae, Hepatitis A virus, Hepatitis B virus, Hepatitis C virus Hepatitis D virus, Hepatitis E virus, Herpes simplex virus type 1, Herpes simplex Virus type 2, Histoplasma capsulatum, hookworm, American hookworm, Influenza Lactobacillus casei, Boca human virus, Ehrlichia ewingii ngii), Anaplasma phagocytophilum, human metapneumovirus, Yale Richter chaffeensis, human papillomavirus, human parainfluenza virus , dwarf tapeworm, diminutive tapeworm, Epstein-Barr virus, Orthomyxoviridae, Iso Isospora belli, Kingell a kingae), Klebsiella pneumoniae, Klebsiella ozena, Klebsiella rhinosierrero Klebsiellarhinoscleromotis, Klebsiella purpurea Lassa fever virus, Legionella pneumophila, Legionella pneumophila, Mycobacterium leprosum, Mycobacterium lepromatosis, and Mycobacterium lepromatosis erium lepromatosis), Leptospira, Listeria, Borreliosis and other Borrelia species, Wuchereria bancrofti and Brugia malayi, Lymphocytic choriomeningitis virus Rus (LCMV), Plasmodium genus, Marble Virus, measles virus, Burkholderia pseudomallei lei), Neisseria meningitidis, Fluke yokogawai, Microsporidia, Molluscum contagiosum virus (MCV), mumps Viruses, Rickettsia typhi, Mycoplasma pneumoniae, various bacteria (Actinomyces Flea mycetoma) and fungi (mycetoma), parasitic fly larvae of the Diptera order, Chlamydia trachomatis Neisseria gonorrhoeae, vCJD prion, Nocardia asteroides and other Nocardia species, Onchocerca volvulus, Blastomyces brasiliensis, Paragonimus and other Paragonimus species, Pasteurella species, Body lice, body lice, Phthiruspubis, whooping cough Bacteria, Yersinia pestis, Streptococcus pneumoniae, Pneumocystis cysticercosis, Poliovirus, Prevotella spp., Neisseria gonorrhoeae Gleria amoeba, JC virus, Chlamydia psittacosis, Coxiella burnetii, Rabies virus, bead-chain E. coli and rat bite fever spirochetes, respiratory RS virus virus, Rhinosporidium sayberi, Rhinovirus, Rickettsia spp., Rickettsia mites , Rift Valley fever virus, Rocky Mountain spotted fever rickettsia, rotavirus, rubella virus , Salmonella spp., Atypical pneumonia coronavirus, Scabies mites, Schistosoma spp., Shigella spp., Varicella zoster Herpes virus, Variola major or minor, Sporothrix schenckii, Staphylococcus aureus, Yellow Staphylococcus aureus, Streptococcus pyogenes, Strongyloides stercoralis, Syphilis spirochete, Cestodes spp., Clostridium tetani, Ringworm, Avian Cophyton tonsurans, Tinea, Epidermophyton floccosum, Trichophyton rubrum and Trichophyton mentagrophytes, Trichophyton rubrum, Holtea werneckii, Ringworm, Malassezia, Roundworm, Roundworm, Toxoplasma gondii, Trichinella spiralis, Trichomonas vaginalis, Whipworm, Mycobacterium tuberculosis, Tula hot flora Ureaplasma urealyticum, Venezuelan equine encephalitis virus, cholera Bacillus, Guanarito virus, West Nile virus, Trichosporonium spp., Yersinia pseudotuberculosis, Yersinia enterocolitica , Yellow fever virus, Mucorales (mucormycosis) and insect mesh mold (entomophthorasis), Pseudomonas aeruginosa, Campylobacter fetus (Vibrio), Aeromonas bacteria, Edwardsiella spp. tarda, Yersinia pestis, Shigella dysenteriae, Shigella sonnei, Salmonella typhimurium, Treponema esculentum pertenue, Treponema carateneum, Fensenburgdorferi, Borrelia bullii Gudolferi, Leptospira hemorrhagic jaundice, Pneumocystis carinii, Brucella abortus, Porcine Produced bacteria, Malta fever bacillus, Mycoplasma spp., Rickettsia typhi, Rickettsia tsutsugamushi , Chlamydia spp., pathogenic fungi (Aspergillus fumigatus, Candida albicans , Histoplasma capsulatum); protozoa (Entamoeba histolytica, Trichomonas vaginalis, Trichomonas humilis) Solanum, Trypanosoma gambiens, Trypanosoma rhodesiense, Leishmania donovani A, Leishmania tropicalis, Leishmania brasiliensis, Pneumocystis carinii pneumonia, 3 days Plasmodium falciparum, Plasmodium falciparum, malignant malaria); or helminths (Schistosoma japonicum, Man These include, but are not limited to, Schistosoma haematobium, Schistosoma haematobium, and hookworms.

[0211] Other antibodies as cell-binding ligands for use in the present invention for the treatment of viral diseases Examples of antibodies include, but are not limited to, antibodies against pathogenic virus antigens, and the pathogenic virus Examples of viruses include, but are not limited to, viruses from the family Poxyirida e), Herpesviridae, Adenoviridae, Papovaviridae, Enteroviridae , Picornaviridae, Parvoviridae, Reoviridae, Retroviridae, Influenza influenza virus, parainfluenza virus, mumps, measles, respiratory syncytial Viruses, rubella, arboviruses, rhabdoviruses, arenaviridae, non-A / non-B hepatitis inflammation viruses, rhinoviruses, coronaviruses, rotaviruses, tumor viruses [e.g. HBV (hepatocellular carcinoma), HPV (cervical cancer, anal cancer), Kaposi's sarcoma-associated herpes virus ( Kaposi's sarcoma), EB virus (nasopharyngeal carcinoma, Burkitt's lymphoma, primary central nervous system lymphoma tumor), MCPyV (Merkel cell carcinoma), SV40 (Simian virus 40), HCV (hepatitis C virus) caused by viruses such as HTLV-I (adult T-cell leukemia / lymphoma); Immune diseases: [e.g., human immunodeficiency virus (AIDS)], CNS viruses: [ For example, JCV (progressive multifocal leukoencephalopathy), MeV (subacute sclerosing panencephalitis), LCV (liver vein leukoencephalopathy), Lymphocytic choriomeningitis), arboviral encephalitis, Orthomyxoviridae (presumed) (Encephalitis lethargica), RV (Rabies), Vesicular stomatitis, Herpes virus meningitis, Ramsay Hunt syndrome type II; poliovirus (acute poliomyelitis, post-polio syndrome), HTLV- I (tropical spastic paralysis)]; cytomegalovirus (CMV) retinitis, HSV (herpes simplex virus) Cardiovascular diseases (e.g., CBV (pericarditis, myocarditis)); respiratory / acute nasopharyngeal Headache / viral pneumonia: [EB virus (EBV infection / infectious mononucleosis), cytomegalovirus (HBV) Rovirus, SARS coronavirus (Severe Acute Respiratory Syndrome), Orthomyxovirus Influenza virus A / B / C (influenza / avian influenza) ), Paramyxovirus: Human parainfluenza virus (parainfluenza), R SV (human respiratory syncytial virus), hMPV]; digestive system viruses [MuV (mumps) esophagitis), cytomegalovirus (CMV esophagitis); adenovirus (adenovirus infection) Infection); Rotavirus, Norovirus, Astrovirus, Coronavirus, HBV (type B) Hepatitis virus), CBV, HAV (hepatitis A virus), HCV (hepatitis C virus), HDV (hepatitis D virus), HEV (hepatitis E virus), HGV (hepatitis G virus) )]; urogenital viruses [e.g., BK virus, MuV (mumps)] It can be enjoyed.

[0212] According to a further object, the present invention also provides a method for preparing a conjugate of the present invention in a pharmaceutical composition comprising a pharmaceutically acceptable carrier, diluent, or the like. The present invention relates to a pharmaceutical composition for treating cancer or an autoimmune disease, comprising the compound of formula (I) together with an agent or excipient. Methods for treating cancer, infectious diseases, and autoimmune diseases include in vitro ), in vivo, or ex vivo. An example of an in vitro therapy would be to express all but the desired mutants that do not express the target antigen. or to kill mutants expressing undesired antigens. Examples of ex vivo therapy include hematopoiesis prior to transplantation (HSCT). The stem cells (HSCs) are treated and then transferred to the same patient's body to kill diseased or malignant cells. For example, prior to autologous transplantation in the treatment of cancer and autoimmune diseases. Transplantation to remove cancer or lymphocyte cells from the marrow or to prevent graft-versus-host disease A method for removing T cells and other lymphoid cells from allogeneic bone marrow or tissue prior to transplantation. Clinical ex vivo treatment can be performed by: Removing bone marrow from a patient or other individual. After harvesting the cells, the conjugate of the present invention was added at a concentration ranging from 1 pM to 0.1 mM. The cells are cultured in a serum-containing medium at 37°C for 30 minutes to approximately 48 hours. The time (= dose) can be easily determined by an experienced clinician. After the culture is completed, the bone marrow cells The cells are washed with serum-containing medium and returned to the body by known methods such as intravenous injection. Between the acquisition and reinfusion treatments, the patient may receive other treatments (e.g., ablative chemotherapy or total body irradiation). If so, the processed bone marrow cells are cryopreserved in liquid nitrogen using standard medical equipment. can be.

[0213] Chemotherapeutic / cytotoxic agents for synergistic effects

[0214] Chemotherapeutic agents that can be used synergistically with the present invention include cytotoxic agents. A "small molecule drug," as the term is used broadly herein, is a small molecule drug, e.g. , organic, inorganic, or For a better definition of small molecule drugs, see WO05058367 A2 and U.S. Pat. No. 4,956,303, as well as other documents, These drugs include known drugs and potential drugs. This includes things that are

[0215] Known drugs include, but are not limited to:

[0216] (1) Chemotherapeutic agents:

[0217] a) an alkylating agent selected from: nitrogen mustards; chlorambucil; Chlornaphazine, cyclophosphamide, dacarbazine, estramustine, ifosfa Mido, mechlorethamine, mechlorethamine oxide hydrochloride, mannomustine, mitobronite ol, melphalan, mitolactol, pipobroman, nobembrine, fenesterine, Prednimustine, thiotepa, trofosfamide, uracil mustard; CC-1065 and synthetic analogues of adozelesin, carzelesin and bizelesin; duocarmycin and and its synthetic analogues KW-2189 and CBI-TMI; benzodiazepine dimers or pyrrolobenzodiazepine (PBD) dimer, tomaymycin dimer, indolinobene zodiazepine dimer, imidazobenzothiadiazepine dimer, or oxazolidinone Diazepine dimers; carmustine, lomustine, chlorozotocin, fotemustine, nimethicone nitrosoureas, including sulphonate, ranimustine; busulphan, treosulphan, ibuprofen; Alkyl sulfonates, including muprosulfan and piposulfan; triazenes or dazolines Carbazine; a platinum-containing compound including carboplatin, cisplatin, and oxaliplatin Aziridines, benzodopa, carboquone, metholedopa, and uredopa; ethylenediamine amines, as well as altretamine, triethylenemelamine, triethylenephosphoramide, Triethylenethiophosphoramine, trimethylolmelamine methylmelamines, including methylmelamine;

[0218] b) Plant alkaloids: Vincristine, vinblastine, vindesine, vinorelbine Vinca alkaloids, including navelbine; taxanes, including paclitaxel and docetaxel; Pseudomonas and their analogues: DM1, DM2, DM3, DM4, DM5, DM6, D Maytansinoids, including M7, maytansine, and ansamitocins, and their analogs Cryptophycins, including the cryptophycin 1 and cryptophycin 8 groups; Epothilone erytherobin, discodermolide, bryostatins, dolostatins, -ristatins, tubulysins, cephalostatins; pancratistatin; sarcostatin Dictyoline;Spongistatin, etc.;

[0219] c) DNA topoisomerase inhibitors: 9-aminocamptothecin, camptothecin, Lisnatol, daunomycin, etoposide, etoposide phosphate, irinotecan, mitoxin Santrone, Novantrone, retinoic acid (retinols), teniposide, topotecan, Epipodophyllins, including 9-nitrocamptothecin or RFS 2000; and mites mycins and their analogues;

[0220] d) Antimetabolites: {[Antifolates: (DHFR inhibitors: methotrexate, trimetrexate ester, denopterin, pteropterin, aminopterin (4-aminopteroic acid), includes other folic acid analogs); IMP dehydrogenase inhibitors (mycophenolic acid, tiazofurin, ribavirin, EICAR, etc.); ribonucleotide reductase inhibitors ( hydroxyurea, deferoxamine.)]; [Pyrimidine analogs: uracil analogs body (ancitabine, azacitidine, 6-azauridine, capecitabine, carmofur, Tarabin, dideoxyuridine, doxifluridine, enocitabine, 5-fluorouracil fluoxuridine, floxuridine, raltitrexed); cytosine analogs: (cytarabine, Cytosine arabinoside, fludarabine; Purine analogs: (azathioprine, fludarabine) Contains: ludarabine, mercaptopurine, thiamiprine, thioguanine); folinic acid folic acid supplements} etc.;

[0221] e) Hormonal therapy agents: receptor antagonists: [Antiestrogens: (megestrol, raloxifene LHRH agonists: (goserelin, leuprorelin acetate) Antiandrogens: (including bicalutamide, flutamide, calcitriol, propionamide) dromostanolone phosphate, epithiostanol, goserelin, leuprolide, mepitios acetaminophen, nilutamide, testolactone, trilostane, and other similar androgen inhibitors Retinoids / deltoid: [Vitamin D3 analogs: (CB1093, EB1 089, KH1060, cholecalciferol, ergocalciferol); light Dynamic therapy agents: (verteporfin, phthalocyanine, photosensitizer Pc4, demethoxyhypo Cytokines: (interferon alpha, interferon gamma, tumor tumor necrosis factor (TNF), including TNF domain-containing human proteins)]}, etc.;

[0222] f) Kinase inhibitors: BIBW2992 (anti-EGFR / Erb2), imatinib, GEF ritinib, pegaptanib, sorafenib, dasatinib, sunitinib, erlotinib, nilotinib tinib, lapatinib, axitinib, pazopanib, vandetanib, E7080 (anti-VEG FR2), mubritinib, ponatinib, bafetinib, bosutinib, cabozantinib, bilirubin Sumodegib, iniparib, ruxolitinib, CYT387, axitinib, tivozanib, Sorafenib, bevacizumab, cetuximab, trastuzumab, ranibizumab, panitum Mab, ispinesib, etc.;

[0223] g) Olaparib, Niraparib, Iniparib, Talazoparib, Veliparib, CEP972 2 (Cephalon), E7016 (Eisai), BGB-290 (Beigene), or 3- Poly(ADP-ribose) polymerase selected from the group consisting of aminobenzamide ( PARP inhibitors, etc.

[0224] h) Enediyne antibiotics (calicheamicins, calicheamicin γ1, δ1, α1 , or β1; dynemicins, including dynemicin A and deoxydynemicin; Esperami Synthin, kedarcidin, C-1027, maduro-peptin, or neocarzinostatin chromo chromophores and related enediyne antibiotic chromophores. , aclacinomycins, actinomycin, anthramycin n), azaserine, bleomycins, cactinomycin , carabicin, carminomycin, carzinophilin; chromoma Isins, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo -L-norleucine, doxorubicin, morpholino-doxorubicin, cyanomorpholino -doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin, epilepsy Bicine, eribulin, esorubicin, idarubicin, marcelomycin, mitomycin nitomycins, mycophenolic acid, nogalamycin, olivomycins, Peplomycin, potfilomycin, puromycin, querramycin, rodrubicin streptonigrin, streptozocin, tuberculin Zin, ubenimex, zinostatin, zorubicin, etc.;

[0225] i) Others: Polyketides (acetogenins), especially bullatacin and bullatacinone; Mucitabine, epoxomicins (e.g., carfilzomib), bortezomib, thalidomide Ido, lenalidomide, pomalidomide, tosedostat, zibrestat, PLX40 32, STA-9090, Stimuvax, Allovectin-7, The Xegeva, Provenge, Elvoy, neurostatin inhibitors (lovastami), ion, cell cycle inhibitors (staurosporine, etc.), 1-methyl-4-phenylpyridine ion, etc.), actinomycins (actinomycin D, dactinomycin etc.), amanitins , bleomycins (bleomycin A2, bleomycin B2, peplomycin, etc.), Anthracyclines (daunorubicin, doxorubicin (adriamycin), idarubicin) Bicine, epirubicin, pirarubicin, zorubicin, etc.), mitoxantrone, MDR inhibitor drugs (verapamil, etc.), Ca 2+ ATP inhibitors (thapsigargin, etc.), histone deacetylases Metabolic enzyme inhibitors (vorinostat, romidepsin, panobinostat, valproic acid, mocetin) Nostat (MGCD0103), Belinostat, PCI-24781, Entinostat SB939, resminostat, gibinostat, AR-42, CUDC-101 , sulforaphane, trichostatin A, etc.); thapsigargin, celecoxib, glitazoline Anti-adrenal drugs (A) Minoglutethimide, mitotane, trilostoma, etc.); aceglatone; aldophosphamide glycoside; aminolevulinic acid; amsacrine; arabinoside, Bestravsil; Bisantren; Edatrexate; Defofamine; Demecolcine; Dimethicone Aziquone; Elfornithine (DFMO), Elfomitine; Elliptinium acetate; Ethyl Crusid, gallium nitrate, gacytosine, hydroxyurea; ibandronate, lentinan ;Lonidamine;Mitoguazone;Mitoxantrone;Mopidamol;Nitracrine;Pent Statins; Fenamet; Pirarubicin; Podophyllic acid; 2-ethylhydrazide; Pro Carbazine; PSK (registered trademark); Razoxane; Rhizoxin; Sizofiran; Spirogel Manium;Tenuazonic acid;Triazicone;2,2',2''-Trichlorotriethylammonium Trichothecenes (T2 toxin, verrucarin A, roridin A, and anguidine, etc.) ); urethane, siRNA, antisense drugs, and nucleolytic enzymes, etc.;

[0226] (2) Anti-autoimmune disease drugs:

[0227] Cyclosporine, cyclosporine A, aminocaproic acid, azathioprine, bromocriptine acetaminophen, chlorambucil, chloroquine, cyclophosphamide, corticoids (e.g., Amcinonide, betamethasone, budesonide, hydrocortisone, flunisolide, fluticasone propionate, fluocortolone danazol), dexamethasone, triamcinolone acetonide, dipropionate (including clomethasone), DHEA, etanercept, hydroxychloroquine, infliximab Ximab, meloxicam, methotrexate, mofetil, mycophenolate, prednisolone including but not limited to Zon, sirolimus, and tacrolimus;

[0228] (3) Anti-infective agents, including but not limited to:

[0229] a) Aminoglycosides: Amikacin, Astromicin, Gentamicin (Netilma Isin, Sisomycin, Isepamicin), Hygromycin B, Kanamycin (Amica cin, arbekacin, bekanamycin, dibekacin, tobramycin), neomycin ( framycin, paromomycin, ribostamycin), netilmicin, spectinomycin Syn, streptomycin, tobramycin, verdamycin;

[0230] b) Amphenicols: azidamphenicol, chloramphenicol, florphen Nicole, thiamphenicol;

[0231] c) Ansamycins: geldanamycin, herbimycin;

[0232] d) Carbapenems: biapenem, doripenem, ertapenem, imipenem, cilasta tin, meropenem, panipenem;

[0233] e) Cephems: Carbacephem (loracarbef), cephacetrile, cefaclor, Cephradine, cefadroxil, cephalonium, cephaloridine, cephalothin or cephalosporin Falotin, cephalexin, cephaloglycin, cefamandole, cephapirin, Fatrizine, cefazaflur, cefazedone, cefazolin, cefbuperazone, cefcape Cefdaloxime, cefepime, cefminox, cefoxitin, cefprozil, Floxadine, ceftezol, cefuroxime, cefixime, cefdinir, cefditoren, Cefepime, cefetamet, cefmenoxime, cefodizime, cefonicid, cefopera Zon, ceforanide, cefotaxime, cefotiam, cefozopran, cephalexin , cefpimizole, cefpiramide, cefpirome, cefpodoxime, cefprozil, cefqui Nom, cefsulodin, ceftazidime, cefteram, ceftibuten, ceftiolene, cefti Zoxime, ceftobiprole, ceftriaxone, cefuroxime, cefuzonam, cephalamycin cefoxitin (including cefoxitin, cefotetan, and cefmetazole), oxacephem (f romoxef, latamoxef);

[0234] f) Glycopeptides: bleomycin, vancomycin (oritavancin, telavancin) ), teicoplanin (dalbavancin), ramoplanin;

[0235] g) Glycylcyclines: Tigecycline;

[0236] h) β-lactamase inhibitors: penams (sulbactam, tazobactam), clavams (clavams) labranic acid);

[0237] i) Lincosamides: clindamycin, lincomycin;

[0238] j) Lipopeptides: Daptomycin, A54145, calcium-dependent antibiotics (C D.A.);

[0239] k) Macrolides: azithromycin, cethromycin, clarithromycin, zirconium Thromycin, erythromycin, flurithromycin, josamycin, ketolide ( Telithromycin, cethromycin), midecamycin, miocamicin, oleandoma Rifamycin (rifampicin, rifampin, rifabutin, rifampin) chin), rokitamycin, roxithromycin, spectinomycin, spiramycin, Tacrolimus (FK506), troleandomycin, telithromycin;

[0240] l) Monobactams: aztreonam, tigemonam;

[0241] m) Oxazolidinones: Linezolid;

[0242] n) Penicillins: Amoxicillin, Ampicillin, Pivampicillin, Hetacillin, Campicillin, methampicillin, talampicillin, azidocillin, azlocillin, benzyl benzathine benzylpenicillin, benzathine phenoxymethylpenicillin , clometocillin, procaine benzylpenicillin, carbenicillin (calindacillin) , cloxacillin, dicloxacillin, epicillin, flucloxacillin, mecillinam ( pivmecillinam), mezlocillin, methicillin, nafcillin, oxacillin, penamecillin penicillin, phenethicillin, phenoxymethylpenicillin, piperacillin, propyl Cillin, sulbenicillin, temocillin, ticarcillin;

[0243] o) Polypeptides: bacitracin, colistin, polymyxin B;

[0244] p) Quinolones: alatrofloxacin, balofloxacin, ciprofloxacin, Linafloxacin, danofloxacin, difloxacin, enoxacin, enrofloxacin Syn, Floxacin, Garenoxacin, Gatifloxacin, Gemifloxacin, Grepaflo flucloxacin, carbafloxacin, levofloxacin, lomefloxacin, marbofloxacin fluoxacin, moxifloxacin, nadifloxacin, norfloxacin, orbifloxacin ofloxacin, pefloxacin, trovafloxacin, grepafloxacin, Tafloxacin, sparfloxacin, temafloxacin, tosufloxacin, torovaf Loxacin;

[0245] q) Streptogramins: Pristinamycin, quinupristin / dalfopristin hmm;

[0246] r) Sulfonamides: Mafenide, Prontosil, Sulfacetamide, Sulfamethoxazole azole, sulfanilamide, sulfasalazine, sulfisoxazole, trimethoprim , trimethoprim-sulfamethoxazole (co-trimoxazole);

[0247] s)Steroidal antibacterial drugs: selected from fusidic acid;

[0248] t) Tetracyclines: doxycycline, chlortetracycline, chromocycline Phosphorus, demeclocycline, lymecycline, meclocycline, methacycline, minocycline Cycline, oxytetracycline, penimepicycline, rolitetracycline, tetracycline tracycline, glycylcyclines (including tigecycline);

[0249] u) Other antibiotics: annonacin, arsphenamine, bactoprenol inhibitors ( Bacitracin), DADAL / AR inhibitor (cycloserine), dictyostatin, Scodermolide, eleutherobin, epothilone, ethambutol, etoposide, faro Penem, fusidic acid, furazolidone, isoniazid, laulimalide, metronidazole, Mupirocin, mycolactone, NAM synthesis inhibitor (fosfomycin), nitrofurantoin paclitaxel, platensimycin, pyrazinamide, quinupristin / dalhopristin stin, rifampicin (rifampin), tazobactamcinidazole, uvaricin;

[0250] (5) Radioisotopes for radiation therapy. Examples of radioisotopes (radionuclides) include: 3 H, 11 C. 14 C. 18 F, 32 P, 35 S, 64 Cu, 68 Ga, 86 Y, 99 T c. 111 In, 123 I, 124 I, 125 I, 131 I, 133 Xe, 177 Lu, 211 At, and 213 Radiolabeled antibodies are used for receptor-targeted imaging. It is very useful in experiments, or for example in the invention of antibody-drug conjugates (Wu et al. (2005) Nature Biotechnology 23(9):1137-1146) for direct comparative targeted therapy. The cell-binding molecules, e.g., antibodies, can be bound to the cell by the cross-linkers of the present invention, as described above. They can be labeled by binding, chelation, or other complex radioisotope-metal binding. This labeling technique is described in Current Protocols in Immunology, Volumes 1 and 2, Coligen et al. I, Ed. Wiley-Interscience, New York, NY, Pubs. (1991). Chelating agents capable of forming metal complexes include DOTA, DOTP, DOTMA, DTPA, and and TETA (Macrocyclics, Dallas, Tex.).

[0251] (6) Another cell-binding molecule-drug conjugate as a synergistic therapy. A preferred synergistic conjugate is -buricin analogues, maytansinoid analogues, taxanoid (taxane) analogues, CC- 1065 analogues, daunorubicin and doxorubicin compounds, amatoxin analogues, benzo Diazepine dimers (e.g., pyrrolobenzodiazepines (PBDs), tomaymycin, and thromycin, indolinobenzodiazepine, imidazobenzothiadiazepine, or oxalic acid Dimers of sazolidinylbenzodiazepines, calicheamicins, and enediyne antibiotic compounds substances, actinomycin, azaserine, bleomycin, epirubicin, tamaxifen, Idarubicin, statin, statin, statin, statin, statin, statin, statin statins auristatin 2-AQ, 6-AQ, EB (AEB), and EFP (AEF) P), duocarmycin, geldanamycin, methotrexate, thiotepa, binde Shin, vincristine, hemiasterlin, nasmamide, microginine, radiosmin, Alterobactin, microscleramine, N-159682, and their analogues and The derivative may be a conjugate with a cytotoxic agent.

[0252] 7) Pharmaceutically acceptable salts, acids, or derivatives of any of the above drugs.

[0253] In yet another embodiment, the immunotoxin is conjugated to a cell-binding molecule as a synergist. The immunotoxins herein are typically diphtheria toxin (DT), cholera toxin (CT), etc. , trichosanthin (TCS), diacin, pseudomonas exotoxin A (ETA'), red blood cells Toxins derived from bacteria or plant proteins, such as diphtheria toxin, AB toxin, and type III exotoxin It is a macromolecular drug that is a cytotoxic protein that requires proteolytic processing for activation. The protoxin may be a highly toxic bacterial pore-forming protoxin of interest. Examples of such protoxins include protoxins The two main components are loaerolysin and its recombinant form, topsalisin. Topsalisin is a modified It is a recombinant protein designed to be selectively activated by prostate enzymes. This induces localized cell death and tissue destruction without damaging adjacent tissues or nerves. vinegar.

[0254] Another synergistic immunotherapy involves checkpoint inhibitors, TCR (T cell receptor) T cells, or CAR (chimeric antigen receptor) T cells, or B cell receptor (BCR), natural killer - (NK) cells, or cytotoxic antibody cells, or anti-CD3, CD4, CD8, CD16 ( FcγRIII), CD27, CD40, CD40L, CD45RA, CD45RO, C D56, CD57, CD57 bright, TNFβ, Fas ligand, MHC class I The molecules (HLA-A, B, C) or NKR-P1 are combined with the conjugates of this patent for synergistic therapy. It is preferable to use them together.

[0255] Prescription and Application

[0256] The conjugates of the patent application may be formulated into a liquid or lyophilized and then reconstituted into a liquid formulation. The conjugate in a liquid formulation or in a formulated lyophilized powder is suitable for formulation. It may account for 0.01% to 99% by weight as a major component in the Conjugate active ingredients were prepared at concentrations of 0.1 g / L to 300 g / L for delivery to patients without antibody aggregation. The liquid formulation containing the active ingredient contains one or more polyols (e.g., sugars), a pH of 4.5 to 7.5, buffers, surfactants (e.g., polysorbate 20 or 80), antioxidants (e.g., corbic acid and / or methionine), tonicity agents (e.g., mannitol, sorbitol or NaCl), chelating agents such as EDTA, metal complexes (e.g., Zn-protein complexes); biodegradable polymers such as polyesters; preservatives (e.g., benzyl alcohol), and / or It may contain free amino acids.

[0257] Buffers suitable for use in the formulation include, but are not limited to, citric acid, ascorbic acid , sodium or potassium gluconate, carbonate, tartarate, succinate, acetate, or phthalate; Organic acid salts such as ammonium or trihydroxyethylamino salts; tris, tromethamine These include hydrochloride, sulfate, or phosphate buffers. Additionally, amino acid cationic components may also be used as buffers. Such amino acid components include arginine, glycine, glycylglycine, Arginine buffers include, but are not limited to, acetate, and histidine. Arginine, arginine chloride, arginine phosphate, arginine sulfate, arginine succinate, etc. In one embodiment, the arginine buffer is arginine acetate. Examples of buffer solutions include histidine chloride-arginine chloride, histidine acetate-arginine acetate, and lysozyme. Histidine phosphate-arginine phosphate, histidine sulfate-arginine sulfate, histidine succinate The pH of the buffer solution is preferably 4.5 to 7.5. In some embodiments, the ratio is about 4.5 to about 6.5, more preferably about 5.0 to about 6.2. In one embodiment, the concentration of the organic acid salt in the buffer is from about 10 mM to about 500 mM.

[0258] The "polyol" optionally included in the formulation is a substance with multiple hydroxyl groups. Polyols are used as stabilizing excipients and / or additives in both liquid and lyophilized formulations. Polyols can be used as tonicity agents to protect biopharmaceuticals from physical and chemical degradation. Preferentially excluded cosolvents can protect the solvent at the protein interface. Increase the effective surface tension, thereby promoting the most energetically favorable structural conformation. The polyols are sugars (reducing and non-reducing sugars) "Reducing sugars" include sugar alcohols and sugar acids. a hemiacetal group that can react covalently with lysine or other amino groups in proteins and "non-reducing sugars" are those that do not have these properties of reducing sugars. Examples of sugars are fructose, mannose, maltose, lactose, arabinose, xylose, Non-reducing sugars include glucose, ribose, rhamnose, galactose, and glucose. Sugars include sucrose, trehalose, sorbose, melezitose, and raffinose. Alcohols include mannitol, xylitol, erythritol, maltitol, and lactitol. erythritol, threitol, sorbitol, and glycerol. Sugar acids include L-gluconate and its metal salts. The polyol content in the sintered dry solids can be 0.0% to 20% by weight. A concentration of 1% to 15% of a non-reducing sugar, sucrose, or trehalose, is selected in the formulation; Trehalose is preferred over sucrose due to the solution stability of trehalose.

[0259] Surfactants optionally included in the formulation include polysorbates (polysorbate 20, polysorbate 21, polysorbate 22, polysorbate 23, polysorbate 24, polysorbate 25, polysorbate 26, polysorbate 27, polysorbate 28, polysorbate 29 ... Polysorbate 40, Polysorbate 65, Polysorbate 80, Polysorbate 81, Polysorbate Benzyl alcohol 85, etc.), poloxamers (e.g., poloxamer 188, poly(ethylene oxy) Poly(propylene oxide), poloxamer 407 or polyethylene-polypropylene ethylene glycol, etc.); Triton; Sodium dodecyl sulfate (SDS); Sodium lauryl sulfate sodium; sodium octylglycoside; lauryl, myristyl, linoleyl, or is stearyl sulfobetaine; lauryl, myristyl, linoleyl, or stearyl Lyl-sarcosine; linoleyl-, myristyl-, or cetyl-betaine; lauramide Propyl-, cocamidopropyl-, linoleamidopropyl-, myristamidopropyl- , palmimidopropyl-, or isostearamidopropyl-betaine (e.g., lauroamide myristamidopropyl-, palmidopropyl-, or isostearamidopropyl- Propyl-dimethylamine; sodium methyl cocoyl, or methyl oleyl taurate Sodium; Dodecyl Betaine, Dodecyl Dimethylamine Oxide, Cocamidopropyl Betaine Tine, and cocoamphoglycinate, "MONAQUAT" (trademark) series (e.g. Isostearyl ethylimidonium ethosulfate; Polyethyl glycol, Poly Propylene glycol and copolymers of ethylene and propylene glycol (e.g., Plu ronics, PF68, etc.); Preferred surfactants are selected from polysorbate 2 Polyoxyethylene glycol ethers such as 0, 40, 60, or 80 (Tween 20, 40, 60, or 80) The surfactant is an ethylene sorbitan fatty acid ester. The concentration of the surfactant in the formulation is 0.0% by weight. In certain embodiments, the surfactant concentration ranges from about 0.01 to about 2.0% by weight. % to about 0.2%. In one embodiment, the surfactant concentration is about 0.02%.

[0260] "Preservatives" that are optionally included in the formulation are compounds that essentially reduce bacterial activity. Examples of potential preservatives include octadecyldimethylbenzyl ammonium chloride, hexadecyl methyl ... Samethonium, benzalkonium chloride (alkybenza chloride, where the alkyl group is a long-chain compound) Other types include benzethonium chloride, a mixture of dimethylammonium chloride, diethylammonium chloride, and benzethonium chloride. Preservatives include phenol, aromatic alcohols such as butyl and benzyl alcohol, methyl alkylparabens such as methylparaben or propylparaben, catechol, resorcinol, cyclohexylparaben, Liquid formulations or preparations include 2-pentanol, 3-pentanol, and m-cresol. The preservative content in the lyophilized powder may be 0.0% to 5.0% by weight. The preservative herein is benzyl alcohol.

[0261] Suitable free amino acids as bulk materials or tonicity or osmolality adjusters in formulations Acids include, but are not limited to, arginine, cystine, glycine, lysine, and histidine. , ornithine, isoleucine, leucine, alanine, glycine glutamic acid or asparagus The basic amino acids are selected from arginine, lysine, and / or histidine. When the composition contains histidine, it acts as a buffer and free amino acid. Although histidine buffers can function as both a carboxylic acid and a non-histidine acid, when histidine buffers are used, they typically function as non-histidine acids. The amino acids include D- and / or lysine-free amino acids, such as histidine buffer and lysine. The amino acid may be in either L- or L-form, but is usually in the L-form. The amino acid may be in any stable salt, e.g. It may be present as a hydrochloride salt such as arginine-HCl. The amino acid content in the powder can be 0.0% to 30% by weight.

[0262] The formulation is available in a liquid formulation at a concentration of up to about 5 mg / ml or in a formulated lyophilized powder. Contains methionine, glutathione, and cinnamal as antioxidants at concentrations of 0.0% to 5.0% by weight. The formulation may optionally contain stearate, cystine, or ascorbic acid. up to about 2 mM in the formulation or 0.0% to 0.3% by weight in the formulated lyophilized powder At a minimum, metal chelators such as EDTA, EGTA, etc. may be optionally included.

[0263] The final formulation may be prepared using a buffer adjuster (e.g., HCl, H2SO4, acetic acid, H3PO4, Acids such as enoic acid, or NaOH, KOH, NH4OH, ethanolamine, diethanolamine amine or triethanolamine, sodium phosphate, potassium phosphate, trisodium citrate The desired pH can be adjusted with a base such as thorium or tromethamine. This is necessary to ensure that the formulation has essentially the same osmolality as human blood. Isotonic preparations generally have an osmotic pressure of approximately 250 to 350 mOsm. can be measured, for example, using a vapor pressure or freezing type osmometer. are mannitol, sorbitol, sodium acetate, potassium chloride, sodium phosphate, Generally, the buffer salt is selected from potassium phosphate, trisodium citrate, or NaCl. Both the hydroxybenzoate and the isotonicity agent may comprise up to 30% by weight of the formulation.

[0264] Other excipients that may be useful in either the liquid or lyophilized formulations of the patent application include, e.g., For example, fucose, cellobiose, maltotriose, melibiose, octulose, riboflavin sugar, xylitol, arginine, histidine, glycine, alanine, methionine, glutamic acid, Amino Acid, Lysine, Imidazole, Glycylglycine, Mannosylglycerate, Tri ton X-100, Pluoronic F-127, cellulose, cyclodextrin (2-hydroxypropyl)-β-cyclodextrin, dextran (10, 40 or 70kD), polydextrose, maltodextrin, ficoll, gelatin, Hydroxypropyl methacrylate, sodium phosphate, potassium phosphate, ZnCl2, zinc, zinc oxide Lead, sodium citrate, trisodium citrate, tromethamine, copper, fibronectin , heparin, human serum albumin, protamine, glycerin, glycerol, EDTA, Meta-cresol, benzyl alcohol, phenol, polyhydric alcohol, or polyhydric alcohol Hydrogen in carbohydrates with a carbonyl group reduced to a primary or secondary hydroxyl group This includes morphological forms.

[0265] Other contemplated excipients that may be utilized in the aqueous pharmaceutical compositions of the patent application include, for example, flavoring agents, Antibacterial agents, sweeteners, antioxidants, antistatic agents, lipids such as phospholipids or fatty acids, cholesterol Steroids such as serum albumin (human serum albumin), recombinant human albumin, gelatin These include protein excipients such as tin and casein, and salt-forming counterions such as sodium. These and additional known pharmaceutical excipients and / or additives suitable for use in the formulations of the invention include: For example, “The Handbook of Pharmaceutical Excipients, 4th edition, Rowe et al., Ed. s. , American Pharmaceuticals Association (2003); and Remington: The Science an d Practice of Pharmacy, 21st edition, Gennaro, Ed. , Lippincott Williams & Wilkin s(2005).”, and is well known in the art.

[0266] A pharmaceutical container or receptacle used to hold a pharmaceutical formulation of any of the conjugates of the patent application. The container may be a vial, bottle, pre-filled syringe, pre-filled or auto-filled syringe. Injector syringe. Liquid formulations are packaged in borosilicate vials or soda-lime glass vials. It can be freeze-dried or drum-dried in vials to form a cake or powder. Powders can also be prepared by efficient spray drying and then packaged in vials for storage and distribution. Or it may be packaged in a pharmaceutical container.

[0267] In a further embodiment, the present invention provides a method for preparing a formulation comprising the steps of: (a) lyophilizing a formulation comprising a conjugate, excipients, and a buffer system; (b) adding the lyophilized mixture of step (a) to a reconstitution medium so that the reconstituted formulation is stable; The formulation of step (a) comprises a stabilizer and the bulking agents, salts, and surfactants described above. and one or more excipients selected from the group comprising a preservative. , using some diluted organic acids or water, i.e., sterile water or bacteriostatic water for injection (BWFI) The reconstitution medium can be water, i.e., sterile water, bacteriostatic water for injection (BWFI), or vinegar. Acid, propionic acid, succinic acid, sodium chloride, magnesium chloride, sodium chloride acid about 1% by weight of a solution of arginine, an acidic solution of magnesium chloride, and an acidic solution of arginine; The amount can be selected from 0 to about 250 mM.

[0268] Liquid pharmaceutical formulations of the conjugates of the patent application should exhibit various predefined properties. Proteins / antibodies tend to form soluble and insoluble aggregates during production and storage. One of the main concerns about liquid pharmaceuticals is their stability. Furthermore, various chemical reactions (deaminase, etc.) occur in solution. This can lead to increased levels of degradation products and reduced biological activity. Preferably, the conjugate in either a liquid or lyophilized formulation is The conjugate should exhibit a shelf life of more than 6 months at 25° C. More preferably, the conjugate is or freeze-dried formulation, must demonstrate a shelf life of more than 12 months at 25°C. The most preferred liquid formulations exhibit a shelf life of approximately 24-36 months at 2-8°C, and the most preferred lyophilized formulations The shelf life of the liquid and lyophilized formulations must be approximately 60 months at 2-8°C. Both must exhibit a shelf life of at least two years at -20°C or -70°C.

[0269] In certain embodiments, the formulation is frozen (e.g., at -20°C or -70°C) and thawed. The drug is stable after, for example, 1, 2, or 3 cycles of freezing and thawing. Qualitative and / or quantitative analysis using a variety of different methods, including evaluation of antibody (protein) ratios and aggregate formation. Can be quantitatively assessed (e.g., UV, size exclusion chromatography, turbidity measurements, and / or visual inspection); cation exchange chromatography, image capillary, etc. charge heterogeneity using electrofocusing (icIEF) or capillary zone electrophoresis Assess the viability of the product by amino- or carboxy-terminal sequence analysis; mass spectrometry; or matrix-supported analysis. assisted laser desorption ionization / time-of-flight mass spectrometry (MALDI / TOF MS), or HP LC-MS / MS; SDS-PAGE analysis to compare reduced and intact antibodies. peptide map (e.g., trypsin or LYS-C) analysis; antibody bioactivity or antigen binding Instability includes aggregation, deamination (e.g., Asn deamination), oxidation ( Met oxidation), isomerization (e.g., Asp isomerization), clipping / hydrolysis / fragmentation cation (e.g., hinge region fragmentation), succinimide formation, unpaired cysteines , N-terminal extension, C-terminal processing, differences in glycosylation, and the like.

[0270] The stable conjugates may also be used in pharmaceutical formulations, for example, in antigen binding assays and / or in vitro at least for a given time, e.g., 12 months, as determined by a cytotoxicity assay at The biological activity of the conjugate in the preparation is preferably about 20% of the biological activity exhibited when the pharmaceutical formulation is prepared. It must "maintain its biological activity," preferably within about 10% (within the error of the assay). stomach.

[0271] For in vivo clinical applications, the conjugate drug of the present invention may be administered in the form of a solution or in sterile water for injection. It is provided in the form of a lyophilized solid that can be redissolved in water. An example of a method is as follows: Weekly intravenous bolus of the conjugate for 8 to 54 weeks. The bolus dose is dissolved in 50 to 1000 mL of saline and then injected into the saline solution. Serum albumin can be added (e.g., 0.5-1 ml of concentrated human serum albumin). The drug dosage is approximately 50 μg to 20 mg / kg body weight per week. , intravenous injection (injection dose ranges from 10 μg to 200 mg / kg per injection) for 4 to 54 weeks After the first course of treatment is completed, the patient can receive a second course of treatment. The exact treatment regimen, including vehicle, diluent, dosage, and duration of treatment, will be determined by an experienced surgeon. It can be determined.

[0272] Treating diseases by selectively killing cell populations using in vivo or ex vivo methods Examples include any type of cancer, autoimmune diseases, transplant rejection, and infectious diseases (viruses). (including viruses, bacteria, or parasites).

[0273] Several factors affect the amount of conjugated drug required for the desired biological effect. The characteristics of the compound, the efficacy and bioavailability of the conjugate drug, the type of disease, the patient's race, the patient's The administration schedule and dosage are determined by taking these factors into account. The route is determined.

[0274] Generally, the conjugates of the present invention via the linker are used at a concentration of 0.1 to 10% w / v. It may be a formulation for parenteral administration dissolved in a physiological buffer so as to contain the compound. Typical dosage ranges are 1 μg / kg body weight to 0.1 g / kg body weight per day, preferably The recommended dose range is 0.01 mg / kg to 20 mg / kg of body weight per day, or The preferred drug dosage depends, for example, on the type and stage of progression of the disease or disorder. the overall health of the individual patient, the relative biological activity of the selected drug, and the compound Dosage form, mode of administration (intravenous, intramuscular, or other), and dosage of the drug in the selected mode of administration Pharmacokinetic properties, as well as the rate of administration (single injection or continuous infusion) and schedule of administration (constant The dose depends appropriately on variables such as the frequency of administration over time.

[0275] In yet another embodiment, a therapeutically effective amount of a conjugate of Formula (I) or Formula (III), or Pharmaceutical compositions comprising any of the conjugates described herein may be used in combination with chemotherapeutic agents, radiotherapeutic agents, immunotherapeutic agents, and the like. anti-cancer, autoimmune disorder, anti-infective, or synergistic effects of cancer, autoimmune disease, or infectious disease It can be administered simultaneously with other therapeutic agents, such as other conjugates for certain treatments or prophylaxis. The synergist is preferably selected from one or several of the following drugs: abatacept , abemaciclib, abiraterone acetate, abraxane, acetaminophen / hydrocodone Calquence, aducanumab, adalimumab, ADXS31-142, ADX S-HER2, afatinib dimalate, aldesleukin, alectinib, alemtuz Mab, alitretinoin, ado-trastuzumab emtansine, amphetamine / dexamin Troamphetamine, anastrozole, aripiprazole, anthracyclines, ali Piprazole, atazanavir, atezolizumab, atorvastatin, avelumab, aki Cicabtagene, ciloleucel, axitinib, belinostat, live BCG, bevacizumab , bexarotene, blinatumomab, bortezomib, bosutinib, brentuximab vedoti brigatinib, budesonide, budesonide / formoterol, buprenorphine, and Ditaxel, cabozatinib, cammatinib, capecitabine, carfilzomib, chimeric antibody Carbohydrate receptor-modified T (CAR-T) cells, celecoxib, ceritinib, cetuximab, chidamivir cyclosporine, cinacalcet, crizotinib, cobimetinib, Cosentyx, Crizotinib, Kymriah, dabigatran, dabrafenib, dacarbazine, dacriz Mab, dacomitinib, daptomycin, daratumumab, darbepoetin Lufa, darunavir, dasatinib, denileukin diftitox, denosumab, Depakote , dexlansoprazole, dexmethylphenidate, dexamethasone, Dignica Top cooling system, dinutuximab, doxycycline, duloxetine, Dube Lisib, dalumab, elotuzumab, emtricib / rilpivirine / tenofovir, Disoproxil fumarate, emtricibine / tenofovir / efavirenz, enoxapa Phosphorus, Ensartinib, Enzalutamide, Epoetin alfa, Erlotinib, Esomeprazole Razol, eszopiclone, etanercept, everolimus, exemestane, everolimus Mus, exenatide ER, ezetimibe, ezetimibe / simvastatin, fenofibrillator filgrastim, fingolimod, fluticasone propionate Quasone, fluticasone / salmeterol, fulvestrant, Gadiva, gefitinib, Latiramer, goserelin acetate, icotinib, imatinib, ibritumomab tiusetan, ibritumomab tiusetan, Brutinib, idelalisib, ifosfamide, infliximab, imiquimod, Immu-C yst, immunoBCG, iniparib, insulin aspart, insulin detemir, Slingulargine, insulin lispro, interferon alpha, interferon alpha-1b, interferon alpha-2a, interferon alpha-2b, Interferon beta, interferon beta 1a, interferon beta 1b, Interferon gamma-1a, lapatinib, ipilimumab, ipratropium bromide / saprophylactic Rubutamol, ixazomib, Kanuma, lanreotide acetate, lenalidomide, lenaliamide , lenvatinib mesylate, letrozole, levothyroxine, lidocaine , linezolid, liraglutide, lisdexamfetamine, LN-144, loratinib, Mantine, methylphenidate, metoprolol, Mekinist, melicitabine / rilpivir Phosphate / Tenofovir, Modafinil, Mometasone, Mycidac-C C), necitumumab, neratinib, nilotinib, niraparib, nivolumab, ofatumumab obinutuzumab, olaparib, olmesartan, olmesartan / hydrochlorothiazol-10, octanoic acid, cyclosporine, cyclosporine Zide, omalizumab, omega-3 fatty acid ethyl ester, oncoline, oseltamivir, Simertinib, oxycodone, palbociclib, palivizumab, panitumumab, panobinostat Stats, pazopanib, pembrolizumab, PD-1 antibodies, PD-L1 antibodies, pemetrexed Sedo, pertuzumab, pneumococcal conjugate vaccine, pomalidomide, pregabalin, proscava ProscaVax, propranolol, quetiapine, rabeprazole, Sodium-223 chloride, raloxifene, raltegravir, ramucirumab, ranibizumab, Regorafenib, ribociclib, rituximab, rivaroxaban, romidepsin, rosuva Statins, ruxolitinib, salbutamol, savolitinib, semaglutide, Belamar, sildenafil, siltuximab, sipuleucel-T -T), sitagliptin, sitagliptin / metformin, solifenacin, solanacine b, sonidegib, sorafenib, sunitinib, tacrolimus, taclimus, tadalafil , Tamoxifen, Tafinlar, Talimogene herparepvec (Talimogene laherparepvec), talazoparib, telaprevir, talazoparib, temozolo mide, tensirolimus, tenofovir / emtricitabine, tenofovir disoproxil Fumarate, testosterone gel, thalidomide, TICE BCG, tiotropium bromide , tisagenlecleucel, toremifene, trametinib tinib, trastuzumab, trabectin (ecteinascidin 743), trametinib, Remelimumab, trifluridine / tipiracil, tretinoin, uro-BCG, ustekinumab Mab, valsartan, veliparib, vandetanib, vemurafenib, venetoclax, Vorinostat, dib-aflibercept, and zostavax, and their analogs , derivatives, pharmaceutically acceptable salts, carriers, diluents or excipients, or combinations thereof. height.

[0276] The drugs / cytotoxic agents used in the conjugation via the branched linkers of this patent are The drug / cytotoxic agent may be any analogue and / or derivative of tubulysin that is known to be a cytotoxic agent. Those skilled in the art will appreciate that for each tubulysin described herein, the resulting compound is still It will be readily understood that the specificity and / or activity of the starting compound can be modified in various ways. Those skilled in the art will also recognize that many of these analog or derivative compounds are also suitable for use in the methods described herein. It will be understood that tubulin analogs may be used instead of the tubulin analogs of the present invention. Tubulysin analogs are tubulysin compounds that may not be described in detail. This includes many analogues and derivatives of

[0277] All references cited herein and in the examples below are expressly incorporated by reference in their entirety. are incorporated into the system. [Example]

[0278] The present invention is further illustrated in the following examples, which are not intended to limit the scope of the invention. The cell lines described in the following examples were obtained from the American Type Culture Collection unless otherwise stated. the American College of Microbiology (ATCC), the German Collection of Microbial Cell Cultures (DSMZ), or the Chinese Academy of Sciences. The cells were maintained in culture medium according to the conditions specified by the Shanghai Cell Culture Institute. Unless otherwise noted, all cell culture reagents were provided by Invitrogen. All anhydrous reagents were commercially available. All other reagents and solvents were obtained from commercial suppliers and stored in nitrogen-sealed bottles. Purchasing was performed according to the standard and used without further purification. NMR spectra were obtained using a Bruker 500 NMR spectrometer. The chemical shifts (Δ) were recorded in ppm. The unit of the coupling constant (J) was Hz. Analytical data were obtained using a Waters Acquity UPLC separator and an Acquity TU Acquisition was performed on a Waters Xevo QTOF mass spectrometer equipped with a V detector.

[0279] Example 1: Di-tert-butyl 1,2-bis(2-(tert-butoxy)-2-oxo Synthesis of (isoethyl)hydrazine-1,2-dicarboxylate. [ka]

[0280] Di-tert-butylhydrazine-1,2-dicarboxylate in DMF (150 ml) To the phosphate (8.01 g, 34.4 mmol), NaH (60% in oil, 2.76 g, 68.8 mmol) After stirring at room temperature for 30 minutes, tert-butyl 2-bromoacetate was added. The mixture was stirred overnight and then cooled to room temperature. The mixture was quenched with the addition of 3 mL of EtOAc, concentrated, and washed with 100 mL of EtOAc and water. The mixture was diluted with HCl, separated, and the aqueous layer was extracted with EtOAc (2×50 ml). Dry with 500 ml SO4, filter, evaporate, and purify using SiO2 column chromatography (EtOA Purification with hexane (1:5 to 1:3) gave the title compound as a colorless oil (12 0.98g, 82% yield. MS ESI m / z calculated C 22 H 41 N2O8[M+H] + 461.28, measured value 461.40.

[0281] Example 2: Synthesis of 2,2'-(hydrazine-1,2-diyl)diacetic acid. [ka]

[0282] Di-tert-butyl 1,2-bis(2-(te rt-Butoxy)-2-oxoethyl)hydrazine-1,2-dicarboxylate (6. To the solution (51 g, 14.14 mmol) was added HCl (12 M, 10 ml). The mixture was Stir for 1 minute, dilute with dioxane (20 ml) and toluene (40 ml), evaporate and distill Coevaporation with oxane (20 ml) and toluene (40 ml) to dryness gave the following crude title compound: The product was obtained (2.15 g, 103% yield, ∼93% purity), which was used in the next step without further purification. Used in process. MS ESI m / z calculated C4H9N2O4 [M+H] + 149.05, measured 149.40.

[0283] Example 3: 2,2'-(1,2-bis((E)-3-bromoacryloyl)hydrazine- Synthesis of 1,2-diyldiacetic acid [ka]

[0284] 2,2'-(hydrazine-1,2-diyl)diacetic acid (1.10 g, 7.43 mmol) A mixed solution of THF (50 ml) and NaH2PO4 (0.1 M, 80 ml, pH 6.0) (E)-3-Bromoacryloyl bromide (5.01 g, 23.60 mmol) was added to the solution. The mixture was stirred for 6 h, concentrated, and diluted with HO / CHCN (1:9) containing 3% formic acid. ) to give the title compound (2.35 g, 77% yield, approximately 93%). MS ESI m / z calculated value C 10 H 11 Br2N2O6[M+H] + 412.89, measured 413.50.

[0285] Example 4: 2,2'-(1,2-bis((E)-3-bromoacryloyl)hydrazine- Synthesis of 1,2-diyl)diacetyl chloride [ka]

[0286] 2,2'-(1,2-bis((E)-3-bromoacetic acid) in dichloroethane (15 ml) (1,2-diisopropyl)hydrazine-1,2-diyl)diacetic acid (210 mg, 0.509 mmol) , (COCl)2 (505 mg, 4.01 mmol), followed by 0.040 mL of DMF After stirring at room temperature for 2 hours, the mixture was concentrated and diluted with dichloroethane (2 x 20 ml). Coevaporation with toluene (2 x 15 ml) to dryness gave the crude title product (not stable). (245 mg, 107% yield) was obtained and used in the next step without further purification. MS ESI m / z calculation value C 10 H9Br2Cl2N2O4[M+H] + 448.82, 450.82, 452.82, 454.82, measurement 448. 60, 450.60, 452.60, 454.60.

[0287] Example 5: tert-Butyl 2,8-dioxo-1,5-oxazocane-5-carboxylate Rate synthesis. [ka]

[0288] At 4°C, 3,3'-azanediyldipropanoic acid (10.00 g, 62.08 mmol) In a 1.0 M NaOH solution (300 ml), di-tert-butyl dicarbonate (2 A solution of 2.10 g (101.3 mmol) of the resulting solution in THF (200 ml) was added over 1 hour. After the addition, the mixture was kept stirring for 2 hours at 4° C. The mixture was carefully washed with 0.2 M H3PO4. Acidify to pH 4, concentrate in vacuo, extract with CH2Cl2, dry with Na2SO4 Evaporate and flush eluate with AcOH / MeOH / CH2Cl2 (0.01:1:5). The compound was purified by silica gel chromatography to give 3'-((tert-butoxycarbonyl )Azanediyl)dipropanoic acid 3 was obtained (13.62 g, 84% yield). ESI MS m / z C 11 H 19 NO6[M+H] + Calculated value 262.27, measured value 262.40.

[0289] At 0°C, 3,3'-((tert-butoxycarbonyl)azanediyl)dipropanoic acid (8.0 g, 30.6 mmol) in CH2Cl2 (500 ml) The mixture was stirred at 0°C for 2 hours and then cooled to room temperature. The mixture was stirred for 1 hour. The mixture was filtered through a short SiO column, and the column was filled with EtOAc / CH2Cl2 (1:6 The filtrate was concentrated and triturated with EtOAc / hexane to give the title compound (5 .64g, yield 74%). ESI MS m / z C11 H 17 NO5[M+H] + Calculated value 244.11, measured value 244.30.

[0290] Example 6: Synthesis of 2,5-dioxopyrrolidin-1-ylpropiolate [ka]

[0291] Propylene in CH2Cl2 (150 ml) and DIPEA (5 ml, 28.7 mmol) ol acid (5.00 g, 71.4 mmol), NHS (9.01 g, 78.3 mmol) ), and EDC (20.0 g, 104.1 mmol) were stirred overnight and evaporated to give SiO Purification by column chromatography (EtOAc / hexane 1:4) gave the title compound The product was obtained as a colorless oil (9.30 g, 79% yield). 1 H NMR (500 MHz, CDCl) δ 2.68 (s, 1H), 2.61(s, 4H). MS ESI m / z calculated value C7H5NaNO4[M+Na] + 190.02, measured value 190.20.

[0292] Example 7: Synthesis of tert-butyl 2-propioloylhydrazinecarboxylate [ka]

[0293] Propylene in CH2Cl2 (150 ml) and DIPEA (5 ml, 28.7 mmol) hydrazine carboxylate (5.00 g, 71.4 mmol), tert-butyl hydrazine carboxylate ( 9.45g, 71.5mmol) and EDC (20.0g, 104.1mmol) The mixture was stirred overnight, evaporated and purified by SiO2 column chromatography (EtOAc / hexane 1: 5) to give the title compound as a colorless oil (7.92 g, 84% yield). 1 H NMR (500 MHz, CDCl3) δ8.76 (m, 2H), 2.68 (s, 1H), 1.39 (s, 9H). MS ESI m / z Calculated value C5H 12 NaN2O2[M+Na] + 155.09, measured 155.26.

[0294] Example 8: Synthesis of propiolohydrazide hydrochloride [ka]

[0295] 2-Propioloylhydrazine carbonate dissolved in 1,4-dioxane (12 mL) at 4 °C tert-Butyl carboxylate (4.01 g, 30.35 mmol) was dissolved in 4 ml of HCl (concentrated The mixture was stirred for 30 minutes and then treated with dioxane (30 ml) and toluene (30 ml). The mixture was diluted with 1 ml of methanol in methylene chloride (5% to 10%) and concentrated in vacuo. The crude mixture was purified on silica gel using a mixture of 10% HCl and 1% formic acid to give the title compound (2.11 g, 83% yield). ESI MS m / z C3H5N2O [M+H] + Calculated value 85.03, measured value 85 .30.

[0296] Example 9: Synthesis of Compound 2 [ka]

[0297] At room temperature, in a 10 L reactor, 2,2-diethoxyacetonitrile (1.00 kg, 7. 74 mol, 1.0 equiv.) to (NH4)2S (48% aqueous solution, 1.41 kg, 9.29 mol, 1.2 equiv.) in methanol (6.0 L). The internal temperature rose to 33°C. After stirring overnight, the reaction mixture was concentrated in vacuo and the residue was dissolved in acetic acid. The aqueous layer was taken up in ethyl acetate (5 L) and washed with saturated NaHCO3 solution (4 x 1.0 L). The organic phases were combined and washed with brine (3 L) and then extracted with ethyl acetate (5 x 1.0 L). The resulting solid was collected by vacuum filtration and diluted with petroleum ether. The filtrate was concentrated and triturated with petroleum ether to give a few grains of white or pale yellow solid. All the products were combined to give the desired product (1.1 kg, 87% yield). 1 HN MR(500 MHz, CDCl3)δ7.81(d, J = 71.1 Hz, 2H), 5.03(s, 1H), 3.73(dq, J = 9 .4, 7.1 Hz, 2H), 3.64(dq, J = 9.4, 7.0 Hz, 2H), 1.25(t, J = 7.1 Hz, 6H).

[0298] Example 10: Synthesis of Compound 3 [ka]

[0299] In a 5 L 3-neck round bottle flask equipped with a reflux condenser and an addition funnel, add 3 L EtOH. Molecular sieves (3A, 500 g) and thioamide (350 g, 2.14 mol , 1.0 equiv.) was added to a mixture of ethyl bromopyruvate (80% purity, 404 mL, 2.5 7 mol, 1.2 eq.) was added over 30 minutes. During the addition, the internal temperature rose slightly. The reaction mixture was then heated to reflux and stirred for 30 minutes. After cooling to room temperature, The reaction mixture was filtered through Celite and the filter cake was washed with ethyl acetate. The two batches of crude product were combined and mixed with silica gel (1.5 kg). , loaded onto a silica gel (10 kg pack) column and separated using ethyl acetate / petroleum ether (10- Elution with 20% HCl gave the thiazole carboxylate as a brown oil (509 g, yield rate 92%).

[0300] Example 11: Synthesis of Compound 4 [ka]

[0301] A solution of acetal (300 g, 1.16 mol) in acetone (3.0 L) was heated to reflux. Then 4N HCl (250 mL) was added to the refluxing solution over 1.0 hour. TLC analysis showed , indicating complete consumption of the starting material. The reaction mixture was concentrated under reduced pressure and the phases were separated. The organic phase Dilute with ethyl acetate (1.5 L) and add saturated NaHCO3 solution (1.0 L), water (1.0 The aqueous phase was washed with water (1.0 L) and brine (1.0 L) and dried over anhydrous Na2SO4. The extracts were combined and extracted with ethyl acetate. The extracts were combined and dried over anhydrous Na2SO4. The solution was filtered and concentrated under reduced pressure. The crude product was extracted with petroleum ether and diethyl ether (5: The resulting solid was collected by vacuum filtration and diluted with petroleum ether and ethyl acetate (1 The filtrate was concentrated and purified by filtration using 0-15% ethyl acetate / petroleum ether. Chromatography gave another crop of the desired product. The combined body weight was 40 g (43% yield). 1 H NMR (500 MHz, CDCl) δ 10.0 8-10.06(m, 1H), 8.53-8.50(m, 1H), 4.49(q, J = 7.1 Hz, 2H), 1.44(t, J = 7.1 Hz, 3H). MS ESI m / z calculated value C7H8NO3S [M+H] + Measurement 186.01;

[0302] Example 12: Synthesis of Compound 6 [ka]

[0303] Dissolve NaN3 (740 g, 11.4 mol) in water (2.0 L) and dichloromethane ( 2.0 L) and cooled to 0 °C, and TfO (700 mL, 4.10 mol, 1.8 equiv. ) was added over 1.5 hours. After the addition was complete, the reaction was stirred at 0°C for 3 hours. The organic phases were separated and the aqueous phase was extracted with dichloromethane (2 x 500 mL). The triflyl azide was washed with saturated NaHCO3 solution (3 x 1.0 L) at room temperature. To the chloromethane solution, add (L)-isoleu in water (3.0 L) and methanol (3.0 L). Synthon (300 g, 2.28 mol, 1.0 equiv.), K2CO3 (472 g, 3.42 mol l, 1.5 eq.), CuSO4·5H2O (5.7 g, 22.8 mmol, 0.01 eq. The mixture was then added to the flask. During the addition, the internal temperature rose slightly. The mixture was then heated to room temperature for 16 The organic solvent was removed under reduced pressure and the aqueous phase was purified with concentrated HCl (approximately 280 mL added). Acidify to pH 6.5 and dilute with phosphate buffer (0.25 M, pH 6.2, 6.0 L) The solution was then diluted with EtOAc (6 x 2.0 L) to remove the by-product sulfonamide. The mixture was acidified to pH 3 with an additional 400 mL of EtOAc and extracted with EtOAc (4 x 2.0 L). The combined organic layer was washed with brine (2.0 L), dried over anhydrous Na2SO4, filtered, and concentrated. Condensation gave product 6 as a pale yellow oil (320 g, 89% yield). 1 H NMR (500 MHz) , CDCl3)δ12.01(s, 1H), 3.82(d, J = 5.9 Hz, 1H), 2.00(ddd, J = 10.6, 8.6, 5.5 Hz, 1H), 1.54(dqd, J = 14.8, 7.5, 4.4 Hz, 1H), 1.36-1.24(m, 1H), 1.08-0 .99(m, 3H), 0.97-0.87(m, 3H).

[0304] Example 13: Synthesis of Compound 10 [ka]

[0305] (S)-2-methylpropane-2-sulfinamide (100 g, 0. 825 mol, 1.0 equiv.) in THF (1 L), Ti(OEt)4 (345 mL, 1.82 mol, 2.2 eq) and 3-methyl-2-butanone (81 mL, 0.825 m ol, 1.0 equiv.) was added. The reaction mixture was refluxed for 16 h, then cooled to room temperature and poured into ice water ( 1 L). The mixture was filtered and the filter cake was washed with EtOAc. The organic layer was The residue was separated, dried over anhydrous Na2SO4, and concentrated to give a residue, which was then subjected to vacuum distillation (15-20 torr, 95°C) to give the product 10 as a yellow oil (141 g, 90% yield). %). 1 H NMR (500 MHz, CDCl3) δ2.54-2.44(m, 1H), 2.25(s, 3H), 1.17(s, 9H) , 1.06(dd, J = 6.9, 5.1 Hz, 6H). MS ESI m / z calculation value C9H 19 NaNOS [M+Na]+ 212.12; measurement Fixed value 212.11.

[0306] Example 14: Synthesis of Compound 11 [ka]

[0307] Diisopropylamine (264 mL, 1.87 mol, 1.65 (equivalent) in dry THF (1 L) with n-butyllithium (2.5 M, 681 mL, 1. The reaction mixture was warmed to 0°C over 30 minutes and then cooled to -7°C. The mixture was cooled to 8° C. Compound 10 (258 g, 1.36 mol, 1.2 equivalents) was added, and THF The reaction mixture was stirred for 1 h, then rinsed with Cl in THF (1.05 L). Ti(OiPr)3 (834 g, 3.17 mol, 2.8 equivalents) was added dropwise. Stirring was continued for 1 hour. After that, compound 4 (210 g, 1.13 mol, 1.0 mL) was dissolved in THF (500 mL). (equivalent) was added dropwise over approximately 1 hour, and the resulting reaction mixture was stirred for 3 hours. Completion of the reaction was confirmed by TLC. Analysis showed that the reaction was carried out with a mixture of acetic acid and THF (v / v 1:1, 300 mL). Quenched, then poured into brine (2 L) and extracted with EtOAc (8 x 1 L). The residue was washed with water and brine, dried over anhydrous Na2SO4, filtered and concentrated. Purification by column chromatography (DCM / EtOAc / PE 2:1:2) gave Compound 11 was obtained as a colorless oil (298 g, 74% yield). 1 H NMR (500 MHz, CDCl3) δ8.13(s, 1H), 6.63(d, J = 8.2 Hz, 1H), 5.20-5.11(m, 1H), 4.43(q, J = 7.0 Hz, 2H), 3.42-3.28(m, 2H), 2.89(dt, J = 13.1, 6.5 Hz, 1H), 1.42(t, J = 7. 1Hz, 3H), 1.33(s, 9H), 1.25-1.22(m, 6H). MS ESI m / z calculation value C 16 H 26 NaN2O4S2[M+ Na] + 397.13, measured value 397.11.

[0308] Example 15: Synthesis of Compound 12 [ka]

[0309] Compound 11 (509 g, 1.35 mol, 1.0 equiv.) dissolved in THF (200 mL) A solution of Ti(OEt)4 (570 mL, 2.72 mol, 2. After the addition was complete, the mixture was stirred for 1 hour, and then NaBH4 (5 1.3 g, 1.36 mol, 1.0 eq) was added in portions over 90 minutes. The mixture was stirred for 3 hours at −78° C. TLC analysis showed that starting material was still present. EtOH (50 mL) was added slowly and the reaction was stirred for 1.5 h, then brine ( The mixture was poured into a 2 L flask containing 250 mL of HOAc and allowed to warm to room temperature. After filtering through Celite, The organic phase was separated, washed with water and brine, dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by column chromatography (EtOAc / PE 1:1) to give Compound 12 was obtained as a white solid (364 g, 71% yield). 1 H NMR (500 MHz, CDCl3 )δ8.10(s, 1H), 5.51(d, J = 5.8 Hz, 1H), 5.23-5.15(m, 1H), 4.41(q, J = 7 .0 Hz, 2H), 3.48-3.40(m, 1H), 3.37(d, J = 8.3 Hz, 1H), 2.29(t, J = 13.0 Hz , 1H), 1.95-1.87(m, 1H), 1.73-1.67(m, 1H), 1.40(t, J = 7.1 Hz, 3H), 1.29 (s, 9H), 0.93(d, J = 7.3 Hz, 3H), 0.90(d, J = 7.2 Hz, 3H). MS ESI m / z calculation Value C 16 H 28 NaN2O4S2[M+Na] + 399.15, measured value 399.14.

[0310] Example 16: Synthesis of Compound 13 [ka]

[0311] Compound 12 (600 g, 1.60 mol, 1.0 eq.) in ethanol (590 mL) To a solution of 4N HCl in dioxane (590 mL) was added slowly at 0°C. The reaction was allowed to warm to room temperature and stirred for 2.5 hours. A white precipitate broke up and was collected by filtration. The filtrate was concentrated and triturated with EtOAc. Two white solids were obtained. The combined products weighed 446 g (90% yield).

[0312] Example 17: Synthesis of Compound 14 [ka]

[0313] Compound 10: Azido-Ile-OH (6, 153 g, 0.97 mol, 2.0 equiv.) Dissolved in THF (1.5 L), cooled to 0 °C, and NMM (214 mL, 1.94 mol, 4 0.0 equiv.) and isobutyl chloroformate (95 mL, 0.73 mol, 2.0 equiv.) sequentially. The reaction was stirred at 0°C for 1.0 h. Compound 13 (150 g, 0.49 mmol) was added. After stirring at 0°C for 30 minutes, the reaction was warmed to room temperature and 2 The reaction was quenched by adding water at 0°C, and the resulting mixture was extracted three times with EtOAc. The combined organic layers were washed with 1N HCl, saturated NaHCO3, and brine. The residue was purified by column chromatography (0- Purification with 30% EtOAc / PE) gave a white solid (140 g, 70% yield). 1 HN MR(500 MHz, CDCl3)δ8.14(s, 1H), 6.57(d, J = 8.9 Hz, 1H), 4.91(d, J = 11. 1 Hz, 1H), 4.44(dd, J = 13.2, 6.3 Hz, 2H), 4.08-3.95(m, 2H), 2.21(dd, J = 24.4, 11.5 Hz, 2H), 1.90-1.79(m, 3H), 1.42(t, J = 6.6 Hz, 3H), 1.37-1.27( m, 2H), 1.11(d, J = 6.4 Hz, 3H), 1.01-0.94(m, 9H). MS ESI m / z calculation value C 18 H 30 N5 O4S [M+H] + 412.19, Measurement 412.19.

[0314] Example 18: Synthesis of Compound 15 [ka]

[0315] Compound 11: Compound 14 (436 g, 1.05 mol, 1.0 equiv.) in CH Into 50 mL of Cl2 solution, imidazole (94 g, 1.37 mmol, 1.3 equiv.) followed by chlorotriethylsilane (222 mL, 1.32 mol, 1.25 equiv.) The reaction mixture was allowed to warm to room temperature over 1 hour. It was then stirred for an additional 1 hour. Brine was added to the reaction mixture. The mixture was added, the organic layer was separated, and the aqueous layer was extracted with EtOAc. The combined organic phases were dried. The mixture was filtered, concentrated under reduced pressure, and then column-eluted with a gradient of 15-35% EtOAc in petroleum ether. Purification by chromatography gave the product 15 as a colorless oil (557.4 g, Yield 95%). 1 H NMR (500 MHz, CDCl3) δ8.12 (s, 1H), 6.75 (d, J = 8.0 Hz, 1H) , 5.20-5.12(m, 1H), 4.44(q, J = 7.0 Hz, 2H), 4.06-3.97(m, 1H), 3.87(d, J = 3.8 Hz, 1H), 2.14(d, J = 3.8 Hz, 1H), 2.01-1.91(m, 3H), 1.42(t, J = 7.1 Hz, 3H), 1.34-1.25(m, 2H), 1.06(d, J = 6.8 Hz, 3H), 1.00-0.93(m, 18H), 0 .88(dd, J = 19.1, 6.8 Hz, 6H). MS ESI m / z calculation value C 24 H 44 N5O4SSi [M+H] + 526.28, measurement Fixed price 526.28.

[0316] Example 19: Synthesis of Compound 16 [ka]

[0317] 15 (408 g, 0.77 mol, 1.0 equiv.) and methyl iodide (145 m L, 2.32 mol, 3.0 equiv) in THF (4 L) was added sodium hydride (in mineral oil 60% dispersion, 62.2 g, 1.55 mol, 2.0 equiv.) was added to the resulting mixture. The mixture was stirred overnight at 0°C and then cooled in ice-water with vigorous stirring to give saturated ammonium chloride. (5 L). The mixture was then extracted with EtOAc (3 x 500 mL) and the organic layer was dried. The mixture was dried, filtered, concentrated, and column chromatographed with a gradient of 15-35% EtOAc in petroleum ether. Purification by chromatography gave the product 16 as a pale yellow oil (388 g, 93 %yield). 1 H NMR (500 MHz, CDCl3)δ8.09(s, 1H), 4.95(d, J = 6.6 Hz, 1H), 4. 41(q, J = 7.1 Hz, 2H), 3.56(d, J = 9.5 Hz, 1H), 2.98(s, 3H), 2.27-2.06(m , 4H), 1.83-1.70(m, 2H), 1.41(t, J = 7.2 Hz, 3H), 1.29(ddd, J = 8.9, 6.8, 1.6 Hz, 3H), 1.01(d, J = 6.6 Hz, 3H), 0.96(dt, J = 8.0, 2.9 Hz, 15H), 0.92 (d, J = 6.6 Hz, 3H), 0.90(d, J = 6.7 Hz, 3H). MS ESI m / z calculation value C 25 H 46 N5O4SSi [M+H] + 540.30, Measurement 540.30.

[0318] Example 20: Synthesis of Compound 17 [ka]

[0319] Compound 16 (1.01 g, 1.87 mmol) in methanol (15 mL) was added to a neutral 0.1N HCl was added dropwise until the pH reached 1.0. Pd / C (10 wt%, 583 mg) After adding the water, the mixture was stirred under H2 (1 atm) at room temperature for 16 h. Then, the Pd / C was filtered off. The filtrate was concentrated under reduced pressure to remove the solids, and the filter pad was washed with methanol. The fraction was redissolved in EtOAc (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated. Compound 17 was obtained as a pale yellow oil (900 mg, 94% yield).

[0320] Example 21: Synthesis of Compound 22 [ka]

[0321] D-pipecolic acid (10.0 g, 77.4 mmol, 1.0 equiv.) in methanol ( 100 mL) of formaldehyde (37% aqueous solution, 30.8 mL, 154.8 mmol) , 2.0 equiv.), followed by Pd / C (10 wt%, 1.0 g). The reaction mixture was heated to 100°C for 1 hour. (1 atm) overnight, and then the filter pad was washed with methanol while the mixture was filtered through Celite. The filtrate was concentrated under reduced pressure to give compound 22 as a white solid (10.0 g, yield: rate 90%).

[0322] Example 22: Synthesis of Compound 23 [ka]

[0323] DN-methylpipecolic acid (2.65 g, 18.5 mmol) in EtOAc (5 0 mL), pentafluorophenol (3.75 g, 20.4 mmol) and DCC ( The reaction mixture was stirred at room temperature for 16 hours, and then The filtrate was filtered through Celite, and the filter pad was washed with 10 mL of EtOAc. Used immediately without purification or concentration.

[0324] Example 23: Synthesis of Compound 28 [ka]

[0325] 2-Amino-2-methylpropanoic acid (500 g, 4.85 mol, 1.0 equiv.), Aqueous methylaldehyde (37%, 1.0 L, 12.1 mol, 2.5 equiv.) and formic acid (1 The mixture was heated to reflux at 80° C. for 3.0 hours. Then, 6N HCl (850 ml) was added. The reaction mixture was concentrated, and the resulting solid was dissolved in ethyl acetate (1.0 L). The solid was collected by filtration, washing three times. The solid was dissolved in water (1.5 L) and washed with 4N NaOH (approx. The solution was concentrated and co-evaporated with ethanol (2.0 L). The remaining water was removed by evaporation. MeOH (2.0 L) was added to the residue and the solid (NaCl) The filtrate was concentrated under reduced pressure to give 639.2 g of a white solid. This contained some NaCl and was used without further treatment.

[0326] Example 24: Synthesis of Compound 29 [ka]

[0327] 2-(Dimethylamino)-2-methylpropanoic acid (97 g, 0.74 mol) in EtO To the Ac solution (1 L), pentafluorophenol (163 g, 0.88 mol) and DI C (126 mL, 0.81 mol) was added, and the reaction mixture was stirred at room temperature for 24 hours. Then, the mixture was filtered through Celite. The filter pad was washed with 10 mL of EtOAc. was used immediately without further purification or concentration.

[0328] Example 25: Synthesis of Compound 30 [ka]

[0329] Dry Pd / C (10 wt%, 300 mg) and azide compound 16 (3.33 g, 6.6 1 mmol) was added to the pentafluorophenyl ester 23 in EtOAc. The mixture was stirred under a hydrogen atmosphere for 27 h, and then the filter pad was washed with EtOAc. The combined organic portions were concentrated and diluted with EtOAc. Purify by column chromatography using a gradient of 0-5% methanol to obtain compound 30 was delivered (3.90 g, 86% yield). MS ESI m / z calculated C 32 H 59 N4O5SSi [M+H] + 6 39.39, measured value 639.39.

[0330] Example 26: Synthesis of Compound 31 [ka]

[0331] The coupled product compound 30 (3.90 g, 6.1 mmol) was dissolved in AcOH / water / T The mixture was dissolved in HF (v / v / v 3:1:1, 100 mL) and stirred at room temperature for 48 hours. The reaction was concentrated and purified by column chromatography (2:98 to 15:85 MeOH / Et OAc) to give compound 31 (2.50 g, 72% yield over two steps). MS ESI m / z Calculated value C 26 H 45 N4O5S [M+H] + 525.30, measured value 525.33.

[0332] Example 27: Synthesis of Compound 32 [ka]

[0333] A solution of compound 31 (2.50 g, 4.76 mmol, 1.0 equiv.) in dioxane at 0° C. (47.7 mL) with LiOH (0.4 N, 47.7 mL, 19.1 mmol, 4.0 equivalents) Aqueous solution of 100 ml of ethyl acetate (amount) was added. The reaction mixture was stirred at room temperature for 2 hours and then concentrated. HPLC (100% CH2Cl2 to CH2Cl2 / MeOH / NH4OH 80: 20:1) to give compound 32 as an amorphous solid (2.36 g, 99% yield) ). MS ESI m / z calculation value C 24 H 41 N4O5S [M+H] + 497.27, measured value 497.28.

[0334] Example 28: Synthesis of Compound 33 [ka]

[0335] Compound 32 (2.36 g, 4.75 mmol) in a pyridine solution (50 mL) at 0 °C Acetic anhydride (2.25 mL, 24 mmol) was added slowly. The reaction mixture was stirred for over 2 hours. The reaction was then warmed to room temperature and stirred at room temperature for 24 hours. The reaction was concentrated and then diluted with dioxane / water (v / v 1:1, 10 mL) for 1 hour to destroy possible anhydrides. After concentration, the residue Column chromatography (100% CH2Cl2 to CH2Cl2 / MeOH / NH 4OH 50:50:1) to give compound 33 as an amorphous white solid ( 2.25g, 88% yield. MS ESI m / z calculated C 26 H 43 N4O6S [M+H] + 539.28, measured value 539.2 8.

[0336] Example 29: Synthesis of Compound 38 [ka]

[0337] To a solution of pentafluorophenyl ester 29 in EtOAc, compound 16 (200 g, 0 0.37 mol) and dry Pd / C (10 wt%, 10 g) were added. The reaction mixture was placed in a hydrogen atmosphere. The mixture was stirred under atmospheric pressure (1 atm) for 27 hours, then filtered through a plug of celite. The filter pad was washed with EtOAc. The combined organic portions were concentrated and washed with 0-5% ethyl acetate in EtOAc. The compound was purified by column chromatography using a gradient of 1000 ml of methanol to give compound 38 (1 84g, 79% yield. MS ESI m / z calculated C 31 H 58 N4O5SSi [M+H] + 627.39, Measurement 6 27.39.

[0338] Example 30: Synthesis of Compound 39 [ka]

[0339] Compound 38 (200g, 0.32mmol) was dissolved in AcOH / water / THF (v / v / v 3 The reaction mixture was concentrated and then stirred at room temperature for 4 days. This step was repeated twice to give compound 39, which was used in the next step. Used directly in the next step. MS ESI m / z calculated value C 25 H 45 N4O5S [M+H] + 513.30, Measurement 513.30.

[0340] Example 31: Synthesis of Compound 40 [ka]

[0341] At 0 °C, a solution of compound 39 (160 g, 0.319 mol, 1.0 equiv.) in MeOH (1 0.2 L) with an aqueous solution of LiOH (0.4 N, 600 mL, 2.55 mol, 8.0 equiv.) The reaction mixture was stirred at room temperature for 2 hours and then concentrated. (pure CH2Cl2 to 80:20:1 CH2Cl2 / MeOH / NH4OH) Compound 40 was obtained as an amorphous solid (140 g, 91% yield over two steps). MS ESI m / z calculation value C 23 H 40 N4O5S [M+H] + 485.27, measured value 485.27.

[0342] Example 32: Synthesis of Compound 41 [ka]

[0343] Compound 27 (143 g, 0.30 mol, 1.0 eq) and DMAP (0.36 g, 2 0.95 mmol, 0.01 equiv) in anhydrous THF (1.4 L) and anhydrous DMF (75 mL) The solution was cooled to 0° C. and added with TEA (82.2 mL, 0.59 mmol, 2.0 equiv.). and acetic anhydride (56 mL, 0.59 mmol, 2.0 equiv.) were added. The reaction mixture was allowed to stand at room temperature. The mixture was heated to rt, stirred for 24 h, and concentrated. / DCM) to give compound 41 as an amorphous solid (147 g, 95% yield). %). MS ESI m / z calculation value C 25 H 44 N4O6S [M+H] + 527.28, Measurement 527.28.

[0344] Example 33: Synthesis of Compound 41a [ka]

[0345] Under N2, a solution of compound 41 (5.0 g, 9.5 mmol, 1.0 equiv.) in anhydrous DCM (1 00 mL), EDC (4.6 g, 23.8 mmol, 2.5 equiv.) and pentafluoro Phenol (4.4 g, 23.8 mmol, 2.5 equiv.) was added at room temperature. The mixture was stirred at room temperature. After stirring at rt for 2 h, the mixture was diluted with DCM (100 mL) and added water (2 x 200 mL) and brine. (200 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and cooled to SiO2 Compound 41a was purified by column chromatography (50% EtOAc / PE) to give a white solid. Obtained as a solid (5.2 g, 79% yield). MS ESI m / z calculated C 31 H 42 F5N4O6S [M+H] + : 69 3.27, measured value 693.27.

[0346] Example 34: Synthesis of Compound 95 [ka]

[0347] In a 500 mL round-bottom flask equipped with a magnetic stir bar, add triphenylphosphine (100 g, 381 mmol, 1.0 equiv) and ethyl 2-bromopropionate (100 mL, 762 The mixture was then heated to 50 °C under a N2 atmosphere overnight. After the white solid (PPh3) dissolved, a large amount of white solid was produced. Trituration with tOAc and filtration gave compound 95 as a white solid (135 g, yield 80%). MS ESI m / z calculation value C 23 H 24 O2P [M-Br] + 363.15, measured value 363.13.

[0348] Example 35: Synthesis of Compound 96 [ka]

[0349] Compound 95 (135.42 g, 305.7 mmol) in dichloromethane (500 ml L) was slowly added to 10% NaOH solution (450 mL) with vigorous stirring. The solution rapidly turned bright yellow. After 30 minutes, TLC analysis indicated the reaction was complete. The layers were separated and the aqueous layer was further extracted with CH2Cl2 (2 x 200 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, and concentrated to give a yellow solid 96. (104 g, 94% yield). MS ESI m / z calculated C 23 H 24 O2P [M+H] + 362.14, measured value 363.1 3. The crude product was used directly in the next step.

[0350] Example 36: Synthesis of Compound 98 [ka]

[0351] Boc-L-Tyr-OMe (670 g, 2.27 mol, 1. 0 equiv.), K2CO3 (358 g, 2.5 mol, 1.1 equiv.), and KI (38 g, 0 To a mixture of 0.227 mol, 0.1 eq) benzyl bromide (283 mL, 2.38 mol (1.05 equiv.) was slowly added. The mixture was then refluxed overnight. Water (6 L) was added. The reaction mixture was extracted with EtOAc (5×100 mL). The combined organic layers were washed with brine ( 2 L), dried over anhydrous Na2SO4, filtered, concentrated, and purified by SiO2 column chromatography. Purification by chromatography (4:1 hexanes / EtOAc) gave a white solid 98 (79 5g, 91% yield. 1 H NMR (500 MHz, CDCl3) δ7.43(d, J = 7.0 Hz, 2H), 7.38(t , J = 7.4 Hz, 2H), 7.32(t, J = 7.2 Hz, 1H), 7.04(d, J = 8.5 Hz, 2H), 6.91( d, J = 8.6 Hz, 2H), 5.04(s, 2H), 4.55(d, J = 6.9 Hz, 1H), 3.71(s, 3H), 3. 03(qd, J = 14.0, 5.8 Hz, 2H), 1.43(s, 9H). MS ESI: m / z: Calculated value C 22 H 28 NO5 [M+H ] + : 386.19, Measurement 386.19.

[0352] Example 37: Synthesis of Compound 99 [ka]

[0353] Ester 98 (380 g, 987 mmol) in anhydrous dichloromethane (1 L) at -78 °C To a solution of 1.0 equiv. (1.0 equiv.), DIBAL (1.0 M in hexane, 2.9 L, 2.9 equiv.) was added. The addition took place over 3 hours. After the addition was complete, the mixture was quenched with 3 L of ethanol. 1N HCl was added dropwise until a pH of 4 was reached. The resulting mixture was warmed to 0°C. The layers were separated. The aqueous layer was separated and further extracted with EtOAc (3 x 3 L). The combined organic solution was washed with brine. The mixture was washed, dried over anhydrous Na2SO4, and concentrated. Trituration with PE / EtOAc and filtration gave A white solid 99 was obtained (263 g, 75% yield). 1 H NMR (500 MHz, CDCl3) δ9. 65(s, 1H), 7.45(d, J = 7.1 Hz, 2H), 7.41(t, J = 7.4 Hz, 2H), 7.35(t, J = 7.1 Hz, 1H), 7.11(d, J = 8.6 Hz, 2H), 6.95(d, J = 8.6 Hz, 2H), 5.07(s, 2H) ), 4.42(dd, J = 12.4, 6.1 Hz, 1H), 3.09(d, J = 6.2 Hz, 2H), 1.46(s, 9H) . MS ESI: m / z calculated value C 21 H 26 NO4[M+H] + : 356.18, measured value 356.19.

[0354] Example 38: Synthesis of Compound 100 [ka]

[0355] Aldehyde 99 (81.4 g, 229 mmol, 1.0 equiv.) was dissolved in anhydrous dichloromethane at room temperature. To a solution of ylide 96 (2 in anhydrous dichloromethane (800 mL) 0.0 equiv.) was added over 30 min. The mixture was stirred at room temperature overnight, then concentrated and cooled to SiO Purification by column chromatography (6:1 petroleum ether / EtOAc) gave a white solid. Solid 100 was obtained (63.4 g, 63% yield). 1 H NMR (500 MHz, CDCl3)δ7.45-7.41 (m, 2H), 7.40-7.35(m, 2H), 7.33(d, J = 7.2 Hz, 1H), 7.10-7.06(m, 2H), 6. 92-6.88(m, 2H), 6.50(dd, J = 8.8, 1.3 Hz, 1H), 5.04(s, 2H), 4.57(s, 2H) , 4.18(q, J = 7.1 Hz, 2H), 2.86(d, J = 8.5 Hz, 1H), 2.72(dd, J = 13.6, 6.8 Hz, 1H), 1.71(d, J = 1.4 Hz, 3H), 1.41(d, J = 2.2 Hz, 9H), 1.28(td, J = 7.5, 5.1 Hz, 4H). MS ESI m / z calculation value C 26 H 33 NaNO5[M+Na] + 462.24, measured value 462.22.

[0356] Example 39: Synthesis of Compound 101 [ka]

[0357] In a hydrogenation bottle, compound 1 in THF (100 mL) and methanol (300 mL) was A solution of 00 (30.2 g, 68.9 mmol) was added to Pd / C (1.83 g, 10 wt%, 50% water) was added. The mixture was shaken under 1 atmosphere of H2 overnight and filtered through Celite (filter aid). Filtration and concentration of the filtrate gave compound 101 as a colorless oil (25.0 g, theoretical yield). . 1HNMR(500 MHz, CDCl3)δ6.99(d, J = 7.0 Hz, 2H), 6.72(d, J = 7.6 Hz, 2H) , 4.39(s, 1H), 4.18-4.04(m, 2H), 3.82(s, 1H), 2.60(dd, J = 37.2, 20.9 Hz , 4H), 1.95-1.81(m, 1H), 1.39(s, 11H), 1.24(dd, J = 9.5, 4.3 Hz, 3H), 1.1 3(t, J = 8.9 Hz, 3H). MS ESI m / z calculation value C 19 H 31 NO5[M+H] + 352.20, measured value 352.19.

[0358] Example 40: Synthesis of Compound 102 [ka]

[0359] Compound 101 (5.96 g, 35.9 mmol, 1.0 equiv.) was dissolved in dichloromethane at room temperature. In a methane solution (200 mL), AcO (3.2 mL, 33.9 mmol, 2.0 equiv.) and HNO3 (65%-68%, 3.5 mL, 50.79 mmol, 3.0 equiv.) The mixture was stirred at room temperature for 30 minutes, and TLC analysis indicated that the reaction was complete. The solution was washed with water (3 x 200 mL) and the aqueous layer was back-extracted with dichloromethane (3 x 100 mL). The combined dichloromethane solution was washed with brine, dried over anhydrous Na2SO4, and Filtration, concentration, and SiO2 column chromatography (5:1 hexane / EtOAc) The compound 102 was purified by HPLC to give compound 102 as a yellow solid (4.18 g, 72% yield). MHz, CDCl3)δ10.49(s, 1H), 7.89(s, 1H), 7.44(d, J = 8.4 Hz, 1H), 7.09(d , J = 8.6 Hz, 1H), 4.32(d, J = 8.3 Hz, 1H), 4.12(dd, J = 14.0, 7.0 Hz, 2H) , 3.80(s, 1H), 2.76(dd, J = 13.0, 6.8 Hz, 2H), 2.59(s, 1H), 1.88 (s, 1H) , 1.37(t, J = 8.7 Hz, 9H), 1.25(dd, J = 13.5, 6.9 Hz, 4H), 1.16(t, J = 8.0 Hz, 3H). MS ESI m / z calculated value C 19 H 28 NaN2O7[M+Na] + 419.19, measured value 419.17.

[0360] Example 41: Synthesis of Compound 103 [ka]

[0361] Ester 102 (1 5.3 g, 38.6 mmol, 1.0 equiv.) to LiOH.HO (16.3 g, 389 A solution of 190 mL of 1,2-dimethyl-2,4-dichloro ... Stir, then dilute with water (400 mL) and add 1N KHSO4 until a pH of 3-4 is reached. After extraction with EtOAc (3 x 300 mL), the organic phase was washed with brine and Aqueous, dried over Na2SO4, filtered and concentrated to give 103 as a yellow solid (14.4 g, theoretical yield). 1 HNMR (500 MHz, CDCl3) δ10.48(s, 1H), 7.98-7.88(m, 1H), 7.42(dd, J = 18.4, 8.2 Hz, 1H), 7.14-7.03(m, 1H), 4.48(d, J = 8.6 Hz, 1H) , 3.90(s, 1H), 2.82-2.53(m, 3H), 1.97-1.82(m, 2H), 1.42-1.27(m, 10H), 1. 21(d, J = 6.7 Hz, 4H). MS ESI m / z calculation value C 17 H 23 N2O7[MH] - 367.16, measured value 367.14.

[0362] Example 42: Synthesis of Compound 104 [ka]

[0363] In a hydrogenation bottle, Pd / C (2.60 g, 10 wt %, 50% water) was added to compound 103 ( 26.0 g, 70.6 mmol, 1.0 equiv.) was added to a methanol solution (260 mL). The mixture was shaken under 1 atmosphere of H2 overnight and then filtered through Celite (filter aid). The filtrate was concentrated to give compound 104 as a green oil (24.0 g, yield >100%). .

[0364] Example 43: Synthesis of Compound 106 [ka]

[0365] tert-Butyl-2-bromopropanoate in dry acetonitrile (1 L) at room temperature methylphenidate (255 g, 1.22 mol, 1.0 equiv.) and triphenylphosphine (320 g, A mixture of 1.22 mol (1.0 equivalent) and acetonitrile was added under reduced pressure. The white precipitate was broken down by adding toluene. The toluene was then decanted. The white solid was dissolved in dichloromethane (1 L) and transferred to a separatory funnel. NaOH (1 L) was added and the organic layer turned yellow immediately upon shaking. The aqueous layer was extracted once with dichloromethane (1 L). After washing once with ethanol (400 mL), it was dried over Na2SO4, filtered and concentrated to give The compound 106 was obtained as a yellow solid (280 g, 58%).

[0366] Example 44: Synthesis of Compound 107 [ka]

[0367] Aldehyde 99 (450 g, 1.27 mol, 1.0 equiv.) was dissolved in dry dichloromethane (3 L), and tert-butyl ester ylide 106 (546 g, 1.40 mmHg) was added thereto. ol, 1.1 equiv.) was added and the solution was stirred at room temperature overnight, at which point the reaction was judged complete by TLC. Purification by column chromatography (10-50% EtOAc / hexane) Compound 107 was obtained as a white solid (444 g, 75% yield). MS ESI m / z calculated C2 8H 38 NO5[M+H] + : 468.27, measured value 468.22.

[0368] Example 45: Synthesis of Compound 108 [ka]

[0369] Compound 107 (63 g, 0.13 mol) was dissolved in methanol (315 mL) and heated at room temperature. The mixture was hydrogenated (1 atm H2) over Pd / C catalyst (10 wt%, 6.3 g) at 40°C overnight. The solvent was filtered off, and the filtrate was concentrated under reduced pressure to give compound 108 (45.8 g, 93% yield). .

[0370] Example 46: Synthesis of Compound 109 [ka]

[0371] A solution of compound 108 (390 g, 1.03 mol, 1.0 equiv.) in THF (4 L) was added at room temperature. ), tert-butyl nitrite (1.06 kg, 10.3 mol, 10 equiv.) was added. The reaction was stirred overnight. After removing the THF, the residue was purified by column chromatography (10 ~50% EtOAc / hexanes) to give compound 109 as a pale yellow solid (314g, 72% yield).

[0372] Example 47: Synthesis of Compound 110 [ka]

[0373] Under nitrogen, a solution of 109 (166 g, 0.392 mol, 1.0 equiv.) in EtOAc (5 To the reaction mixture (00 mL), Pd / C (10 wt%, 16 g) was added, and the reaction flask was evacuated and filled with hydrogen. The reaction mixture was stirred under hydrogen (1 atm) at room temperature for 16 h, then purged three times. Filtration through silica gel and concentration gave the product 110 as a pale yellow foam (146 g, 97% yield) ). 1 H NMR (400 MHz, CDCl3) δ6.62(d, J = 7.9 Hz, 1H), 6.55(s, 1H), 6.43(d , J = 7.3 Hz, 1H), 4.39(dd, J = 53.0, 44.2 Hz, 1H), 3.77(s, 4H), 2.72-2.29 (m, 3H), 1.83-1.58(m, 1H), 1.40(d, J = 7.6 Hz, 18H), 1.24(s, 1H), 1.06( t, J = 5.7 Hz, 3H). MS ESI m / z calculation value C 21 H 35 N2O5[M+H] + 394.25, measured value 395.25.

[0374] Example 48: Synthesis of Compound 114 [ka]

[0375] At −78° C. under N2, (S)-4-isopropyloxazolidin-2-one (5.00 g, 38.7 mmol, 1.0 equiv) in anhydrous THF (200 mL) (2.5 M in hexane, 17.0 mL, 1.2 equivalents) was added and the mixture was stirred for 30 minutes. Stir at -78 °C for 1 h, then add propionyl chloride (4.0 mL, 42.58 mmol, 1 The mixture was stirred at -78°C for an additional hour, and TLC analysis indicated the reaction was complete. Saturated ammonium chloride solution (250 mL) was added and EtOAc (3× The combined organic layer was extracted with 1N NaOH solution (200 mL) and Wash with toluene (300 mL), dry over anhydrous Na2SO4, filter, concentrate, and column Purification by chromatography (7:1 hexane / EtOAc) gave compound 114 as a colorless oil. (6.36 g, 89% yield). MS ESI m / z calculated C9H 16 NO3[M+H] + 186.10, Measurement value: 186.10. 1 H NMR (400 MHz, CDCl3) δ4.48-4.39(m, 1H), 4.27(t, J = 8.7 Hz , 1H), 4.21(dd, J = 9.1, 3.1 Hz, 1H), 3.06-2.82(m, 2H), 2.38(dtd, J = 14. 0, 7.0, 4.0 Hz, 1H), 1.17(t, J = 7.4 Hz, 3H), 0.90(dd, J = 17.0, 7.0 Hz, 6H ).

[0376] Example 49: Synthesis of Compound 115 [ka]

[0377] (S)-4-isopropyl-3-propionyloxazolidine-2- A solution of 2.00 g (11.9 mmol, 1.1 equiv.) of 1H2O3 in anhydrous dichloromethane (20 ml) L), DIPEA (2.3 mL, 12.9 mmol, 1.2 equiv.) and n-BuBO Tf (1.0 M in dichloromethane, 12.0 mL, 1.1 equivalents) was added and mixed. The mixture was stirred at 0°C for 45 min, then cooled to -78°C and the resulting solution of compound 99( The mixture was heated at -78°C for 1 hour. The mixture was stirred at room temperature overnight and then slowly warmed to room temperature. M, pH 7.0, 100 mL) was added. After phase separation, the aqueous phase was further diluted with dichloromethane (3 × 5 The combined organic layers were washed with brine (200 mL) and anhydrous NaCl. The crude product was redissolved in methanol (100 mL). The mixture was then treated with H2O2 (30% aqueous solution, 26 mL, 23 equiv.) at 0 °C for 3 h. Methanol was removed by evaporation and water (100 mL) was added. The resulting mixture was diluted with EtO The combined organic layers were washed with brine (300 mL). , dried over anhydrous Na2SO4, filtered, concentrated, and purified by SiO2 column chromatography ( 3:1 hexanes / EtOAc) to give compound 115 as a foamy solid (2. 70g, 49% yield. 1 H NMR (400 MHz, CDCl3) δ7.52-7.26 (m, 5H), 7.15 (d, J = 7.4 Hz, 2H), 6.93(d, J = 7.3 Hz, 2H), 5.05(s, 2H ), 4.69(d, J = 7.0 Hz, 1 H), 4.47(s, 1H), 4.36(t, J = 7.8 Hz, 1H), 4.17(d, J = 8.5 Hz, 1H), 3.93( d, J = 7.1 Hz, 1H), 3.85(s, 2H), 2.84(d, J = 6.9 Hz, 2H), 2.31(s, 1H), 1. 40-1.37(m, 9H), 1.31(s, 3H), 0.92 (dd, J = 13.4, 6.6 Hz, 6H). MS ESI m / z Calculated value C 30 H 41 N2O7[M+H] + 541.28, measured value 541.30.

[0378] Example 50: Synthesis of Compound 116 [ka]

[0379] Compound 115 (2.50 g, 4.63 mmol, 1.0 equiv) in anhydrous THF (46 mL) amount) and 1,1'-thiocarbonyldiimidazole (2.48 g, 13.89 mmol, A mixture of Et (3.0 equiv.) was refluxed overnight. Water (100 mL) was added and the resulting mixture was diluted with Et The combined organic layers were washed with brine (200 mL). , dried over anhydrous Na2SO4, filtered, concentrated, and purified by SiO2 column chromatography ( Purification with hexanes / EtOAc (3:1) gave compound 116 as a yellow foam (2.3 3g, 77% yield. 1 H NMR (400 MHz, CDCl3) δ8.41(s, 1H), 7.67(s, 1H), 7.36 (dt, J = 16.0, 6.9 Hz, 6H), 7.09(s, 1H), 7.05(d, J = 8.4 Hz, 2H), 6.86(d , J = 8.4 Hz, 2H), 6.32(d, J = 9.5 Hz, 1H), 5.01(s, 2H), 4.56-4.43(m, 2H) , 4.32(ddd, J = 16.2, 15.6, 7.8 Hz, 3H), 4.19(d, J = 8.7 Hz, 1H), 2.96(dd , J = 14.6, 4.4 Hz, 1H), 2.49(dd, J = 14.5, 10.5 Hz, 1H), 2.29(td, J = 13.4 , 6.7 Hz, 1H), 1.31(s, 3H), 1.29(s, 9H), 0.91(dd, J = 13.9, 6.9 Hz, 6H). MS ESI m / z calculated value C 34 H 43 N4O7S[M+H] + 651.27, measured value 651.39.

[0380] Example 51: Synthesis of Compound 117 [ka]

[0381] A solution of compound 116 (1.90 g, 2.92 mmol, 1.0 equiv.) in anhydrous toluene (3 In a 100 mL (1.0 mL) mixture, n-Bu3SnH (1.6 mL, 5.84 mmol, 2.0 equiv.) and azo Diisobutyronitrile (0.05 g, 0.584 mmol, 0.1 equiv) was then added. The mixture was refluxed for 2.5 h, concentrated, and purified by SiO2 column chromatography (5:1 hexane). Purification by hexane / EtOAc gave compound 117 as a white foam (1.21 g, yield 79%). 1H NMR (400 MHz, CDCl3)δ 7.36 (ddd, J = 24.5, 14.5, 7.1 Hz, 5H), 7. 08(d, J = 8.5 Hz, 2H), 6.90(d, J = 8.5 Hz, 2H), 5.04(d, J = 5.1 Hz, 2H), 4.48(d, J = 4.2 Hz, 1H), 4.33(t, J = 8.4 Hz, 1H), 4.22(d, J = 9.7 Hz, 1H) , 4.15(d, J = 8.8 Hz, 1H), 3.81(s, 2H), 2.73(dd, J = 14.1, 5.9 Hz, 1H), 2 .61(dd, J = 14.0, 7.2 Hz, 1H), 2.29(dq, J = 13.5, 6.8 Hz, 1H), 2.11 - 2.00( m, 1H), 1.35(s, 9H), 1.20(d, J = 6.9 Hz, 3H), 0.89(dd, J = 14.0, 6.9 Hz, 6 H). MS ESI m / z calculated value C 30 H 41 N2O6[M+H] + 525.28, measured value 525.37.

[0382] Example 52: Synthesis of Compound 118 [ka]

[0383] Compound 117 (1.20g, 2.29mmol, 1.0eq) in THF solution (30mL ) with an aqueous solution (6 mL) of LiOH (0.192 g, 4.58 mmol, 2.0 equiv.) and and H2O2 (30% aqueous solution, 1.4 mL, 6.0 equiv.) were added. The mixture was stirred at 0°C for 3 hours. After that, the reaction was stopped by adding sodium bisulfite solution (1.5 M, 30 mL). After that, 1N KHSO4 was added dropwise until a pH of 4 was reached. The reaction mixture was then diluted with EtOAc. The EtOAc solution was washed with brine and extracted with anhydrous Na2SO4 The mixture was dried, filtered, concentrated, and purified by column chromatography on SiO2 (3:1 hexane / Et OAc, containing 1% HOAc) to give compound 118 as a white solid (0.7 8g, 82% yield. 1 H NMR (400 MHz, CDCl3) δ7.46-7.28(m, 5H), 7.07(d, J = 7 .7 Hz, 2H), 6.91(d, J = 7.8 Hz, 2H), 4.52(d, J = 8.5 Hz, 1H), 3.87(d, J = 41.8 Hz, 1H), 2.82-2.43(m, 3H), 1.85(t, J = 12.2 Hz, 1H), 1.41(s, 9H), 1. 17(d, J = 6.9 Hz, 3H). MS ESI m / z calculated value C 24 H 32 NO5[M+H] + 414.22, measured value 414.21 .

[0384] Example 53: Synthesis of Compound 119 [ka]

[0385] Compound 118 (0.77 g, 1.86 mmol, 1.0 equiv) in methanol (15 mL) A mixture of 10% ethanol (0.25 g) and Pd / C (10%, 0.25 g) was hydrogenated under 1 atmosphere of H2 for 16 hours. The filtrate was concentrated to give compound 119 as a white solid. Obtained (0.58 g, 96% yield). 1 H NMR (400 MHz, CDCl3) δ7.00(d, J = 7.5 Hz, 2 H), 6.80(s, 2H), 4.51 (d, J = 9.0 Hz, 1H), 3.88(s, 1H), 2.66(dd, J = 65. 6, 22.6 Hz, 4H), 1.88(t, J = 12.2 Hz, 1H), 1.42(s, 9H), 1.14(d, J = 6.6 H z, 3H). MS ESI m / z calculated value C 17 H 26 NO5[M+H] + : 324.17, measured value 324.16.

[0386] Example 54: Synthesis of Compound 120 [ka]

[0387] At 0 °C, a solution of compound 119 (0.57 g, 1.76 mmol, 1.0 equiv.) in THF ( To the resulting solution (10 mL), add t-BuONO (0.63 mL, 5.28 mmol, 3.0 equiv.). The reaction was stirred at 0°C for 1 hour and then at room temperature for 1 hour. Water (50 mL) was added, followed by The reaction mixture was extracted with EtOAc (3×30 mL). The combined organic layers were washed with brine (10 0 mL), dried over anhydrous Na2SO4, filtered, concentrated, and purified by SiO2 column chromatography. The compound was purified by chromatography (2:1 hexane / EtOAc with 1% HOAc). Product 120 was obtained as a yellow solid (0.50 g, 77% yield). 1 H NMR (400 MHz, DMSO) )δ7.92(s, 1H), 7.47(d, J = 8.3 Hz, 1H), 7.05(d, J = 8.5 Hz, 1H), 3.73(s , 1H), 2.78(dd, J = 13.6, 5.3 Hz, 1H), 2.69-2.47(m, 2H), 1.87(t, J = 11.9 Hz, 1H), 1.47-1.37(m, 1H), 1.32 (s, 9H), 1.17(d, J = 7.2 Hz, 3H). MS ESI Calculated m / z value C 17 H 25N2O7[M+H]+ 369.15, measured value 369.14.

[0388] Example 55: Synthesis of Compound 121 [ka]

[0389] Compound 120 (0.50 g, 1.36 mmol, 1.0 equiv) in methanol (10 mL) A mixture of 1000 ppm of ethanol (10 wt %) and Pd / C (0.02 g) was hydrogenated at room temperature for 1 hour (1 atmosphere). The mixture was then filtered through Celite (filter aid). The filtrate was concentrated to give compound 121. Obtained as a white foam (0.43 g, 93% yield). MS ESI m / z calculated C 17 H 27 N2O5[M+H] + 339.18, measured 339.17. 1H NMR (400 MHz, MeOD) δ 6.60 (d, J = 7.9 Hz, 2H), 6.44 (d, J = 7.3 Hz, 1H), 3.71(d, J = 6.3 Hz, 1H), 2.62 - 2.37(m, 3H), 1.83(d dd, J = 13.7, 9.9, 3.7 Hz, 1H), 1.39(s, 9H), 1.13(d, J = 7.1 Hz, 3H).

[0390] Example 56: Synthesis of Compound 124 [ka]

[0391] A solution of maleic anhydride (268 g, 2.73 mol) in acetic acid (1 L) was added to 4-aminobutanol. After stirring at room temperature for 30 minutes, the reaction mixture was heated to 1. Reflux for 5 hours and cool to room temperature. Evaporation in vacuo gave a residue which was taken up in EA and water. and brine, dried over anhydrous Na2SO4, filtered and concentrated. Crystallization from tOAc and PE gave a white solid (400 g, 80% yield). 1 H NMR (500 MHz, CDCl3)δ6.71(s, 2H), 3.60(t, J = 6.7 Hz, 2H), 2.38(t, J = 7.3 H z, 2H), 2.00-1.84(m, 2H).

[0392] Example 57: Synthesis of Compound 125 [ka]

[0393] Compound 124 (400 g, 2.18 mol, 1.0 equiv.) in CHCl (1.5 L) N-hydroxysuccinimide (276 g, 2.40 mmol, 1.1 Equivalents) and DIC (303 g, 2.40 mol, 1.1 equiv.) were added and stirred overnight. The reaction mixture was concentrated and purified by column chromatography (1:2 petroleum ether / EtOAc). Purification gave the NHS ester 125 as a white solid (382 g, 63% yield). 1 HN MR(500 MHz, CDCl3)δ6.74(s, 2H), 3.67(t, J = 6.8 Hz, 2H), 2.85(s, 4H), 2 .68(t, J = 7.5 Hz, 2H), 2.13-2.03(m, 2H).

[0394] Example 58: Synthesis of Compound 126 [ka]

[0395] At 0°C, a solution of compound 124 (60 g, 328 mmol, 1.3 equiv.) in THF (600 mL), NMM (85.3 mL, 984 mmol, 3.0 equiv.) was added with stirring. Isobutyl chloroformate (44.6 mL, 426 mmol, 1.3 equiv) was added dropwise. After stirring at 0°C for 2 hours, the resulting mixture was added with Compound 104 (10 2g, 259mmol, 1.0 equiv) in THF (400mL). After completion, the reaction was stirred for an additional 30 min, then quenched with water (300 mL) and EtO The combined organic layers were dried, filtered, and concentrated to give 9- Compound 12 was obtained by column chromatography using a gradient of 35% EtOAc / PE. 6 was obtained as a pale yellow solid (104 g, 73% yield). 1 H NMR (400 MHz, CDCl3) δ8. 86(s, 1H), 8.40(d, J = 17.3 Hz, 1H), 6.87(s, 3H), 6.70(s, 2H), 4.53-4.16 (m, 0H), 3.79(s, 1H), 3.62(t, J = 6.1 Hz, 1H), 2.63(s, 1H), 2.40(t, J = 6.9 Hz, 1H), 2.12-1.88(m, 4H), 1.84-1.64(m, 1H), 1.38(t, J = 9.6 Hz, 6H) , 1.06(t, J = 6.0 Hz, 3H).

[0396] Example 59: Synthesis of Compound 127 [ka]

[0397] Compound 126 (12.7 g, 22.7 mcg) dissolved in CH2Cl2 (20 mL) at 0 °C mol) was treated with TFA (40 mL) and the reaction was allowed to warm to room temperature and stirred for 3 hours. The residue was triturated with diethyl ether to give a pale yellow toluene solution. Solid 127 was collected (11.4 g, theoretical yield).

[0398] Example 60: Synthesis of Compound 128 [ka]

[0399] To a solution of carboxylic acid 33 (40 mg, 0.074 mmol, 1.0 equiv) in EtOAc, Pentafluorophenol (27 mg, 0.148 mmol, 2.0 equiv.) and DCC ( The reaction mixture was stirred at room temperature for 16 hours. The mixture was then filtered through a pad of Celite, washing the pad with EtOAc. was concentrated, redissolved in DMA (6 mL), and compound 127 (56.6 mg, 0.13 mmol) was obtained. 1) and DIPEA (47.4 μL, 0.18 mmol) were added. The reaction mixture was stirred at room temperature. The mixture was stirred for 24 hours, then concentrated and purified by reverse phase HPLC (C18 column, 10-100% acetone). nitrile / water) to give compound 128 as a white solid (43 mg, yield 6 3%). MS ESI m / z calculated value C 46 H 66 N7O 11 S [M+H] + 924.45, Measurement 924.45.

[0400] Example 61: Synthesis of Compound 132 [ka]

[0401] Compound 41a (11 g, 15.9 mmol, 1.0 equiv.) and Compound 127 ( 12.3 g, 23.8 mmol, 1.5 eq) in DMF (100 mL) A (6.9 mL, 39.7 mmol, 2.5 equiv) was added. The reaction mixture was allowed to warm to room temperature. The mixture was stirred for 1 hour. The mixture was concentrated in vacuo and purified by silica gel column (100% DCM to 10 %MeOH / DCM) to give compound 132 as an amorphous solid (10 g , yield 69%). MS ESI m / z calculated value C 45 H 65 N7O 11 S [M+H] + 912.45, Measurement 912.45.

[0402] Example 62: Synthesis of Compound 166 [ka]

[0403] (R)-4-isopropyloxazolidin-2-one (25.0%) was prepared at −78 °C under N2. g, 0.194 mol, 1.0 equiv.) in anhydrous THF (1150 mL) i (85.0 mL, 0.213 mol, 1.1 equiv.) was added, and the mixture was stirred at the same temperature for 1 h. After stirring for a while, a lot of white solids were formed. Then, at -78 °C, 20.0 mL, 0.232 mol, 1.2 eq) was added and the mixture was stirred at the same temperature for 1 h. After monitoring the consumption of (S)-4-isopropyloxazolidin-2-one by TLC, The solution was poured into saturated ammonium chloride solution (1.2 L) and the mixture was diluted with EA (700 mL, 3 The organic extract was extracted with 1.0 N NaOH solution (1.0 L) and 1.0 N NaOH solution (50 mL x 2). Wash with ethanol (1.0 L), dry over anhydrous sodium sulfate, filter, concentrate in vacuo, and The title compound was purified by iO2 column chromatography (PE:EA=10:1) to give the Obtained as a colored oil (32.6 g, 90.8%). ESI m / z calculated C9H 17 NO3[M+H] + : 186.1, Measurement value: 186.1. 1 H NMR (400 MHz, CDCl3) δ4.48-4.37(m, 1H), 4.27(t, J = 8.7 Hz, 1H), 4.21(dd, J = 9.1, 3.1 Hz, 1H), 3.04-2.82(m, 2H), 2.45-2.30(m, 1H), 1.17(t, J = 7.4 Hz, 3H), 0.90(dd, J = 17.1, 7.0 Hz, 6H).

[0404] Example 63: Synthesis of Compound 167 [ka]

[0405] (R)-4-isopropyl-3-propionyloxazolidine-2- In a solution of 18.4 g (99.5 mmol, 1.1 equiv.) in anhydrous DCM (200 mL) , BuBOTf (1 M dichloromethane solution, 100 mL, 100 mmol, 1.1 equiv. ) and DIPEA (19 mL, 108.6 mmol, 1.2 equiv.) were added and the mixture was The mixture was stirred at -78°C for 45 min. Aldehyde 99 (32.2 g, 90.5 mmol) was obtained at -78°C. A solution of 1.0 equivalents of 1H-dimethylformamide in dichloromethane (320 mL) was added and stirred at the same temperature for 1 hour. The solution was then allowed to warm slowly to room temperature for 15 hours. The mixture was then added to 700 mL of potassium phosphate. The mixture was poured into a buffer solution (pH 7.0) and extracted with ethyl acetate. The organic extract was washed with brine and The residue was dried over anhydrous sodium sulfate, filtered, and then concentrated in vacuo. After cooling to 0°C, 30% H2O2 aqueous solution (225 mL) was slowly added. The mixture was stirred at the same temperature for 3 hours. After adding water (750 mL), the mixture was concentrated in vacuo to give The methanol was removed, and the resulting aqueous solution was diluted with ethyl acetate (500 mL, 150 mL x 2). The organic extract was washed with 5% sodium bicarbonate solution and brine, and then with anhydrous sodium sulfate. The extract was dried over sodium, filtered, concentrated in vacuo, and purified by SiO column chromatography (PE: EA=3:1) to give the title compound as a white foam (31.7 g, 64.8%) . MS ESI m / z calculation value C 30 H 41 N2O7[M+H] + : 541.3, measured value 541.3. 1 H NMR (400 MHz, CDCl3 )δ7.49-7.29(m, 5H), 7.17(t, J = 10.7 Hz, 2H), 6.93(d, J = 7.0 Hz, 2H), 5 .06(s, 2H), 4.28(dd, J = 44.4, 36.4 Hz, 3H), 4.04-3.52(m, 3H), 3.11-2.73 (m, 2H), 2.35(s, 1H), 1.41(t, J = 16.3 Hz, 9H), 0.91( dd, J = 15.6, 6.4Hz , 5H).

[0406] Example 64: Synthesis of Compound 168 [ka]

[0407] Compound 167 (28.3g, 52.3mmol, 1.0 in anhydrous THF (350mL) 1,1-thiocarbonyldiimidazole (TCDI) (35.1 g, 1 57.0 mmol, 3.0 equiv.) was added and the mixture was heated under reflux overnight. After monitoring the reaction rate by TLC, the mixture was concentrated in vacuo and purified by SiO2 column chromatography ( The title compound was purified with PE:EA=3:1 to give the title compound as a pale yellow foam (26.1 g, 7 6.8%). MS ESI m / z calculation value C 34 H 43 N4O7S [M+H] + : 651.3, Measurement 651.3. 1 HNMR (400 MHz, CDCl3)δ8.21(s, 1H), 7.43(d, J = 11.8 Hz, 1H), 7.42-7.28(m, 5H), 7.0 6(d, J = 8.3 Hz, 2H), 7.01(s, 1H), 6.80(d, J = 8.3 Hz, 2H), 6.17(dd, J = 8.5, 2.9 Hz, 1H), 4.96(s, 2H), 4.42-4.04(m, 5H), 2.83(dd, J = 14.2, 6.2H z, 1H), 2.69(dd, J = 14.2, 7.1 Hz, 1H), 2.32(dd, J = 6.8, 4.2 Hz, 1H), 1.3 7(s, 9H), 1.30(d, J = 6.9 Hz, 3H), 0.87(dd, J = 9.9, 7.0 Hz, 6H).

[0408] Example 65: Synthesis of Compound 169 [ka]

[0409] Compound 168 (26.0 g, 40.0 mmol, 1.0 equiv.) in anhydrous toluene under N2 To the solution (350 mL), n-Bu3SnH (21.5 mL, 80.0 mmol, 2.0 equivalents) was added amount) and 2,2'-azobis(2-methylpropionitrile) (AIBN) (0.066 g, 0.01 eq.) was added. The mixture was heated under reflux for 1 h. The consumption of the starting material was confirmed by T After LC monitoring, the mixture was concentrated in vacuo and purified by SiO2 column chromatography (PE: EA=5:1) to give the title compound as a white foam (6.0 g, 37.3%) . MS ESI m / z calculation value C 30 H 41 N2O6[M+H] + : 525.3, Measurement 525.3. 1 HNMR (400 MHz, CDCl3 )δ7.37(ddd, J = 25.1, 15.1, 7.1 Hz, 5H), 7.08(d, J = 7.9 Hz, 2H), 6.89(d , J = 8.4 Hz, 2H), 5.03(s, 2H), 4.61(d, J = 8.4 Hz, 1H), 4.40(s, 1H), 4.3 2-4.08(m, 2H), 3.91-3.66(m, 2H), 2.83(d, J = 8.4 Hz, 1H), 2.60(t, J = 10 .1 Hz, 1H), 2.33(s, 1H), 1.71(s, 1H), 1.41(s, 9H), 1.15(d, J = 6.5 Hz, 3 H), 0.87(dd, J = 17.0, 7.0 Hz, 6H).

[0410] Example 66: Synthesis of Compound 170 [ka]

[0411] Compound 169 (7.84 g, 15 mL) in THF (90 mL) and water (30 mL) at 0 °C. 0.0 mmol, 1.0 equiv.) in 30% aqueous H2O2 solution (11.4 mL, 112.5 mm ol, 7.5 equiv.) in LiOH HO (1.57 g, 37.5 mmol, 2.5 equiv.) ) was added and the mixture was stirred at the same temperature for 3 hours. L) was added at 0°C, and the mixture was stirred at the same temperature for 30 minutes. 4 was added slowly until the pH reached 4. The resulting solution was diluted with EA (200 mL, 75 mL × 2). The organic extract was washed with brine, dried over anhydrous sodium sulfate, filtered, and purified. The mixture was concentrated in vacuo and purified by SiO2 column chromatography (PE:EA=2:1). The title compound was obtained as a white solid (6.18 g, 100%). MS ESI m / z calculated C 24 H 32 N1O5[M+H]+: 414.2, measured value 414.2. 1 HNMR (400 MHz, CDCl3)δ7.39(ddd, J = 24.5, 15 .0, 7.2 Hz, 5H), 7.11(d, J = 7.8 Hz, 2H), 6.93(d, J = 8.3 Hz, 2H), 5.06(s , 2H), 4.44(t, J = 8.3 Hz, 1H), 3.83(d, J = 69.4 Hz, 1H), 2.85-2.61(m, 2H ), 2.61-2.40(m, 1H), 1.99-1.70(m, 1H), 1.39(d, J = 26.1 Hz, 9H), 1.19(s , 3H).

[0412] Example 67: Synthesis of Compound 171 [ka]

[0413] In a hydrogenation bottle, compound 170 (6.18 g, 15.0 mmol, 1.0 equiv.) was added to M To the eOH solution (50 mL) was added Pd / C (0.6 g, 10% Pd / C). The mixture was shaken under 1 atmosphere of hydrogen overnight and then filtered. The filtrate was concentrated to give the title compound. Obtained as a colorless oil (4.8 g, 99% yield). MS ESI m / z calculated C 17 H 26 N1O5[M+H] + : 324 .2, measured value 324.2. 1HNMR (400 MHz, CDCl3) δ6.97(d, J = 6.5 Hz, 2H), 6.74(d, J = 8.2 Hz, 2H), 3.93-3.66(m, 1H), 2.58(tdd, J = 19.5, 12.9, 7.4 Hz, 3H), 1. 75(ddd, J = 20.1, 16.3, 7.7 Hz, 1H), 1.37(d, J = 21.5 Hz, 9H), 1.11(d, J = 7.0 Hz, 3H).

[0414] Example 68: Synthesis of Compound 172 [ka]

[0415] Compound 171 (4.8 g, 15.0 mmol, 1.0 equiv.) was dissolved in anhydrous T t-BuONO (18.0mL, 150mmo...

Claims

1. Side-chain linked conjugate compounds of formula (III): 【Chemistry 7】 During the ceremony, 【Chemistry 2】 represents a single bond; n is 1 to 30; T is a cell binding agent / molecule selected from the group consisting of antibodies, single chain antibodies, antibody fragments that bind to target cells, monoclonal antibodies, single chain monoclonal antibodies, monoclonal antibody fragments that bind to target cells, chimeric antibodies, chimeric antibody fragments that bind to target cells, domain antibodies, domain antibody fragments that bind to target cells, adnectins that mimic antibodies, DARPins, lymphokines, hormones, vitamins, growth factors, colony stimulating factors, nutrient transport molecules (transferrin), and / or cell binding peptides, proteins, small molecules attached to albumin, polymers, dendrimers, liposomes, nanoparticles, vesicles, or (viral) capsids; L １ and L ２ are independently the same or different, 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, or the formula (OCH ２ CH ２ ) ｐ OR ３ , (OCH ２ CH-(CH ３ )) ｐ OR ３ , NH(CH ２ CH ２ O) ｐ R ３ , NH(CH ２ CH (CH ３ ) O) ｐ R ３ , N[(CH ２ CH ２ O) ｐ R ３ ]-[(CH ２ CH ２ O) ｐ’ R ３’ ], (OCH ２ CH ２ ) ｐ COOR ３ , C.H. ２ CH ２ (O.C.H. ２ CH ２ ) ｐ COOR ３ (p and p' are independently selected integers from 0 to 1000), or combinations thereof; １ ~C ８ Alkyl of C ２ ~C ８ Heteroalkyl, alkylcycloalkyl, heterocycloalkyl of C ３ ~C ８ or a natural or unnatural amino acid, or a sequence of identical or different dipeptide, tripeptide, tetrapeptide, pentapeptide, hexapeptide, heptapeptide, octapeptide, nonapeptide, decapeptide, undecapeptide, or dodecapeptide units (Aa). ｒ , r=1-12 (1 to 12 amino acid units); W is C １ ~C １８ is the expansion unit of; w is 1, 2, or 3; V １ and V ２ are independently O, NH, S, C １ ~C ８ Alkyl, C ２ ~C ８ Heteroalkyl, alkenyl, or alkynyl, C ３ ~C ８ aryl, heterocycle, carbocycle, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroaralkyl, heteroalkylcycloalkyl, or alkylcarbonyl, or (Aa)r, where r=1-12 (1-12 amino acid units), and (Aa)r is composed of natural or unnatural amino acids, or dipeptide, tripeptide, tetrapeptide, pentapeptide, hexapeptide, heptapeptide, octapeptide, nonapeptide, decapeptide, undecapeptide, or dodecapeptide units of the same or different sequences, or (CH ２ CH ２ O) ｐ (p=0 to 1000); １ and v ２ is independently 0, 1, or 2, provided that v １ and v ２ are not 0 at the same time; １ Or v ２ When is 0, the side chain Q １ Or Q ２ It means that one of the fragments is absent; Q １ and Q. ２ are independently represented by formula (I-q1): 【Chemistry 3】 During the ceremony, 【Chemistry 4】 L １ Or L ２ is the site of binding to; 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 , N.H.S. (O) ２ N.H., C.H. ２ S (O) ２ NH, OS (O) ２ O, C.H. ２ S (O) ２ O, Ar, ArCH ２ , ArO, ArNH, ArS, ArNR １ Or (Aa)q1; G ３ は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 , N.H.S. (O) ２ (OH), NHS (O) ２ N.H. ２ , C.H. ２ S (O) ２ N.H. ２ , OS(O) ２ OH, OS (O) ２ OR １ , C.H. ２ S (O) ２ OR １ , Ar, ArR １ , ArOH, ArNH ２ , ArSH, ArNHR １ Or (Aa) ｑ１ where (Aa) ｑ１ are peptides containing sequences of identical or different 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, NHC(O)NH; Y ２ O, NH, NR １ , C.H. ２ , S, NHNH, Ar; p １ , p ２ , and p ３ are independently between 0 and 100, but not 0; q １ andq ２ are independently 0 to 24; R １ , R ２ , R ３ , and R ３’ are independently H, C １ ~C ８ Alkyl; C ２ ~C ８ Heteroalkyl or heterocycle; C ３ ~C ８ Ar is aryl, Ar-alkyl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, carbocycle, or alkylcarbonyl; Or W, Q １ , Q ２ , L １ , L ２ , V １ , or V ２ may be independently absent, provided that Q １ and Q. ２ cannot exist simultaneously; D is a tubulysin analog having the following formula (II): or a pharma- ceutically acceptable salt, hydrate, or hydrated salt thereof; or a polymorphic crystal; or an optical isomer, racemate, diastereomer, or enantiomer thereof: 【Chemistry 5】 During the ceremony, 【Chemistry 6】 is a binding site that independently binds to W; In the formula, R １ , R ２ , R ３ , and R ４ are independently H, C １ ~C ８ Alkyl; C ２ ~C ８ Heteroalkyl, or heterocyclic; ３ ~C ８ Aryl, Ar-alkyl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, carbocycle, or alkylcarbonyl; or R １ R ２ , R １ R ３ , R ２ R ３ , R ３ R ４ , R ５ R ６ , R １１ R １２ , or R １３ R １４ form a 3- to 7-membered carbocyclic, cycloalkyl, heterocyclic, heterocycloalkyl, aromatic, or heteroaromatic ring system; R １ and R ２ may be absent independently when bonded to W independently or simultaneously, and Y １ is N or CH; In the formula, R ５ , R ６ , R ８ , and R １０ are independently H or C １ ~C ４ is alkyl or heteroalkyl; In the formula, R ７ are independently H, R １４ , -R １４ C(=O)X １ R １５ or -R １４ X １ R １５ and X １ is O, S, SS, NH, CH ２ , or NR １４ is; In the formula, R ９ is 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 OP(=O)(OR １５ ) selected from In the formula, R １１ are independently H, R １４ , -R １４ C(=O)R １６ , -R １４ X ２ R １６ , -R １４ C(=O)X ２ And X ２ -O-, -S-, -NH-, -N(R １４ ) -, -O-R １４ -, -S-R １４ -, -S(=O)-R １４ -, or -NHR １４ is; In the formula, R １２ is R １５ , -OH, -SH, -NH ２ , N.H., N.H. ２ , -NH(R １５ ), -OR １５ , -R １５ C.O.R. 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 -O-, OH, SH, -S-, NH ２ , -NH-, -N(R １５ ) -, -O-R １５ -, -S-R １５ -, -S(=O)-R １５ -, CH ２ , or -NHR １５ - is; R １３ and R １４ are independently H, O, S, NH, N(R １５ ), NHNH, -OH, -SH, -NH ２ , N.H., N.H. ２ , -NH(R １５ ), -OR １５ , CO, -COX ２ , -COX ２ R １６ , R １７ ,F,Cl,Br,I,SR １６ , N.R. １６ R １７ , N=NR １６ , N=R １６ , NO ２ , SOR １６ R １７ , S.O. ２ R １６ , S.O. ３ R １６ , OSO ３ R １６ , P.R. １６ R １７ , P.O.R. １６ R １７ , P.O. ２ R １６ R １７ , OP(O)(OR １７ ) ２ , O.C.H. ２ OP(O)(OR １７ ) ２ , O.C.(O)R １７ ,OC(O)OP(O)(OR １７ ) ２ , PO(OR １６ ) (OR １７ ), OP(O)(OR １７ ) OP (O) (OR １７ ) ２ , O.C.(O)N.R. １７ -O-(C ４ ~C １２ Glycoside), -N-(C ４ ~C １２ Glycoside); C １ ~C ８ Alkyl, heteroalkyl; ２ ~C ８ Alkenyl, alkynyl, heteroalkyl, heterocycloalkyl of C ３ ~C ８ Ar-alkyl, carbocycle, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl, or an ester, ether, or amide of 2 to 8 carbon atoms; or 1 to 8 amino acids (NH(Aa) １～８ Or CO(Aa) １～８ or a peptide comprising the formula (OCH ２ CH ２ ) ｐ Or (OCH ２ CH (CH ３ )) ｐ (p is an integer from 0 to 1000), or a combination thereof; ２ is O, S, SS, NH, CH ２ , O.H., S.H., and N.H. ２ , CHR １４ , or NR １４ is; R １５ , R １６ , and R １７ are independently H, C １ ~C ８ alkyl, heteroalkyl; ２ ~C ８ Alkenyl, alkynyl, heteroalkyl, heterocycloalkyl of C ３ ~C ８ Ar-alkyl, carbocycle, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl, alkylcarbonyl, or Na ＋ , K ＋ , Cs ＋ , Li ＋ , Ca ２＋ , Mg ＋ , Zn ２＋ , N ＋ (R １ ) (R ２ ) (R ３ ) (R ４ ), H.N. ＋ (C ２ H ５ O.H. ３ It is salt; Y １ and Y ２ is independently N or CH; q is 0 or 1; when q=0, Y ３ does not exist, and Y ４ , Y ５ , Y ６ , and Y ７ are independently CH, N, NH, O, S, or N(R １ ), and therefore, Y ２ , Y ４ , Y ５ , Y ６ , and Y ７ form a heteroaromatic ring of furan, pyrrole, thiophene, thiazole, oxazole, imidazole, pyrazole, triazole, tetrazole, thiadiazole; when q=1, Y ３ , Y ４ , Y ５ , Y ６ , and Y ７ are independently CH or N, so that Y ２ , Y ３ , Y ４ , Y ５ , Y ６ , and Y ７ form the aromatic rings of benzene, pyridine, pyridazine, pyrimidine, pyrazine, triazine, tetrazine, and pentazine.

2. A side chain linked compound of formula (V) capable of reacting with a cell binding agent / molecule T to form a side chain linked conjugate compound of formula (III) according to claim 1: 【Chemistry 10】 During the ceremony, D, W, w, L １ , L ２ , Q １ , Q ２ , V １ , V ２ , v １ , and v ２ is the same as claim 1; In the formula, Lv １ and Lv ２ is independently a reactive group capable of reacting with a thiol, amine, carboxylic acid, selenol, phenol, or hydroxyl group on a cell-binding agent / molecule. １ and Lv 2 are OH, 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.

3. Side Chain Q １ and Q. ２ is independently selected from Iq-01 to Iq-35. 【Chemistry 11】 【change】 During the ceremony, R ２５ and R ２５’ are independently 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 Estelle, 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, C ２ -C １８ Fatty acid, or C ２ -C １８ selected from fatty ammonium lipids; １ and X ２ are independently NH, N(R1'), O, CH ２ , S, C(O), S(O), S(O ２ ), P(O)(OH), NHNH, CH=CH, Ar, or (Aa)q １ is selected from, where q １ = 0-24 (0-24 amino acids, q １ = 0 means absence; X １ , X ２ , X ３ , X ４ , Y １ , Y ２ , and Y ３ are independently NH, N(R1'), 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)q １ where X is selected from １ , X ２ , X ３ , X ４ , Y １ , Y ２ , and Y ３ may not be present independently; １ , p ２ , and p ３ are independently 0 to 100 and not 0; q １ , q ２ , and q ３ are independently 0 to 24; R １’ , R ２’ , R ３’ , and R ４’ are independently H and C １ -C ６ alkyl; Aa is a natural or unnatural amino acid; (Aa)q 1 is the same or different peptide sequence, and q １ = 0 is (Aa)q １ does not exist.

4. The side chain linked conjugate compound of formula (III) according to claim 1 or the side chain linked compound of formula (V) according to claim 2, wherein D (tubulysin structure) is selected from I-01 to I-75 or a pharma- ceutically acceptable salt, hydrate, or hydrate salt thereof; or a polymorphic crystal of these compounds; or an optical isomer, racemate, diastereomer, or enantiomer thereof: 【Chemistry 12】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 During the ceremony, R ２０ is H;C １ ~C ８ Linear or branched alkyl or heteroalkyl of C ２ ~C ８ Linear or branched alkenyl, alkynyl, alkylcycloalkyl, heterocycloalkyl of C ３ ~C ８ Ar-alkyl, heterocycle, carbocycle, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl; carbonate (-C(O)OR １７ ), carbamates (-C(O)NR １７ R １８ or a carboxylate, ester, ether, or amide having 1 to 8 carbon atoms; or an amino acid having 1 to 8 carbon atoms; or a compound of the formula (OCH ２ CH ２ ) ｐ Or (OCH ２ CH (CH ３ )) p where p is an integer from 0 to 1000; or R ２０ is absent and the oxygen forms a ketone, or a combination 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 , O.C.H. ２ OP(O)(OR １８ ) ２ ,OC(O)OP(O)(OR １８ ) ２ , OPO (OR １８ ) ２ , NHPO (OR １８ ) ２ , OP(O)(OR １８ ) OP (O) (OR １８ ) ２ , O.C.(O)R １８ , O.C.(O)N.R. １８ , OSO ２ (OR １８ ), O-(C ４ -C １２ -glycoside), linear or branched alkyl or heteroalkyl; ２ ~C ８ Linear or branched alkenyl, alkynyl, alkylcycloalkyl, heterocycloalkyl of C ３ ~C ８ Ar-alkyl, heterocycle, carbocycle, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl; carbonate (-C(O)OR １７ ), carbamates (-C(O)NR １７ R １８ ) ; R １７ and R １８ are independently H, C １ ~C ８ Linear or branched alkyl or heteroalkyl of C ２ ~C ８ Linear or branched alkenyl, alkynyl, alkylcycloalkyl, heterocycloalkyl of C ３ ~C ８ Ar-alkyl, heterocycle, carbocycle, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl; carbonate, carbamate; R １９ is H, OH, NH ２ , OSO ２ (OR １８ ), X.H.H. 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 of C ２ ~C ８ Alkenyl, alkynyl, alkylcycloalkyl, heterocycloalkyl of C ３ ~C ８ or a pharmaceutical salt; where X is O, S, NH, NHNH, or CH ２ is; R ７ has the same definition as in claim 1.

5. W, L １ , L ２ , V １ , and V ２ are independently composed of one or more linker moieties of the following structure: 【Chemistry 13】 【change】 or an L- or D-, natural or non-natural peptide comprising 1 to 20 identical or different amino acids; 【Chemistry 14】 During the ceremony, 【Chemistry 15】 is the binding site; X ２ , X ３ , X ４ , X ５ , or X ６ are independently NH; NHNH; N(R ３ );N(R ３ ) N (R ３’ );O;S;C １ -C ６ Alkyl of C ２ -C ６ Heteroalkyl, alkylcycloalkyl, heterocycloalkyl of C ３ -C ８ Ar is selected from aryl, Ar-alkyl, heterocycle, carbocycle, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl; or 1 to 8 amino acids. R ３ and R ３’ are independently H; C １ -C ８ Alkyl of C ２ -C ８ Heteroalkyl, alkylcycloalkyl, heterocycloalkyl of C ３ -C ８ Aryl, Ar-alkyl, heterocycle, carbocycle, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl; or an ester, ether, or amide having 1 to 8 carbon atoms; or a group of the formula (OCH ２ CH ２ ) ｐ Or (OCH ２ CH (CH ３ )) ｐ (p is an integer of 0 to 1000), or a combination thereof.

6. W, L １ , L ２ , V １ , and V ２ are independently composed of the following: (A): A self-immolative moiety, a peptide unit, a hydrazone bond, a disulfide, an ester, an oxime, an amide, or a thioether bond. The self-immolative components include 2-aminoimidazole-5-methanol derivatives, heterocyclic PAB analogs, β-glucuronides, and o- or p-aminobenzyl acetals; or any one of the following structures: 【Chemistry 16】 In the formula, ( ＊ Atoms marked with X are points of attachment to other moieties; １ , Y １ , Z ２ , and Z ３ is independently NH, O or S; Z １ are independently H, NHR １ , OR １ , S.R. １ , COX １ R １ And X １ and R １ is defined as above; v is 0 or 1; U １ are independently H, OH, and C １ ~C ６ Alkyl, (OCH ２ CH ２ ) ｎ ,F,Cl,Br,I,OR ５ , S.R. ５ , N.R. ５ R ５’ , N=NR ５ , N=R ５ , N.R. ５ R ５’ , NO ２ , S.O.R. ５ R ５’ , S.O. ２ R ５ , S.O. ３ R ５ , OSO ３ R ５ , P.R. ５ R ５’ , P.O.R. ５ R ５’ , P.O. ２ R ５ R ５’ , OPO (OR ５ ) (OR ５’ ), or OCH ２ PO (OR ５ ) (OR ５’ ) in which R ５ and R ５ ’ are independently H, C １ ~C ８ Alkyl; C ２ ~C ８ alkenyl, alkynyl, heteroalkyl, or amino acid; ３ ~C ８ aryl, heterocyclic, carbocyclic, cycloalkyl, heterocycloalkyl, heteroaralkyl, alkylcarbonyl, or glycoside; or a medicinal cationic salt; (B): A non-self-immolative linker moiety comprising one of the following structures: 【Chemistry 17】 【change】 In the formula, ( ＊ The atom marked with X is the point of attachment; １ , Y １ , U １ , R ５ and R ５’ is as defined above; r is 0 to 100; m and n are independently 0 to 20; (C): at least one bond capable of being broken down under physiological conditions; a releasable linker comprising a pH-labile, acid-labile, base-labile, oxidatively labile, metabolically labile, biochemically labile, or enzymatically labile bond and having one of the following structures: -(CR) ５ R ６ ) ｍ (A) ｒ (CR) ７ R ８ ) ｎ (OCH) ２ CH ２ ) ｔ -、-(CR ５ R ６ ) ｍ (CR) ７ R ８ ) ｎ (A) ｒ (OCH) ２ CH ２ ) ｔ -、-(A]) 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 (A) ｔ -、-(CR ５ R ６ ) ｍ (CR) ７ =R ８ )(CR ９ R １０ ) ｎ (A) ｔ (OCH) ２ CH ２ ) ｒ -、-(CR ５ R ６ ) ｍ (NR) １１ CON ｔ (CR) ９ R １０ ) ｎ (OCH) ２ CH ２ ) ｒ -、-(CR ５ R ６ ) ｍ (A) ｔ (NR) １１ CR) ９ R １０ ) ｎ (OCH) ２ CH ２ ) ｒ -、-(CR ５ R ６ ) ｍ (O) (A) 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 (C) (A) ｔ -(CR) ９ R １０ ) ｎ (OCH) ２ CH ２ ) ｒ -、-(CR ５ R ６ ) ｍ (NR) １１ CON ｔ (CR) ９ R １０ ) ｎ (OCH) ２ CH ２ ) ｒ -、-(CR ５ R ６ ) ｍ (O) (A) 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 (O.C.H. ２ CH ２ ) ｒ -, - (CR ５ R ６ ) ｍ (CO) (Aa) ｔ (CR ９ R １０ ) ｎ (O.C.H. 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 (C.C.R. ７ R ８ ) ｎ -, - (CR ５ R ６ ) ｔ -Thienyl-(CO)(CR ７ R ８ ) ｎ -, - (CR ５ R ６ ) ｔ -imidazolyl-(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-methylpiperazine-(CO)(Aa) ｔ (CR ７ R ８ ) ｎ -, - (CR ５ R) ｍ -(Aa) ｔ Phenyl-, -(CR ５ R ６ ) ｍ -(Aa) ｔ Frill-,-(CR ５ R ６ ) ｍ -Oxazolyl (Aa) ｔ -, - (CR ５ R ６ ) ｍ -Thiazolyl (Aa) ｔ -, - (CR ５ R ６ ) ｍ -Thienyl (Aa) ｔ -, - (CR ５ R ６ ) 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 (A) ｒ (CR) ７ R ８ ) ｎ (OCH) ２ CH ２ ) ｔ -、-K(CR ５ R ６ ) ｍ (CR) ７ R ８ ) ｎ (A) 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 (A) ｔ -、-K(CR ５ R ６ ) ｍ (CR) ７ =R ８ )(CR ９ R １０ ) ｎ (A) ｔ (OCH) ２ CH ２ ) ｒ -、-K(CR ５ R ６ ) ｍ (NR) １１ CON ｔ (CR) ９ R １０ ) ｎ (OCH) ２ CH ２ ) ｒ -、-K(CR ５ R ６ ) ｍ (A) ｔ (NR) １１ CR) ９ R １０ ) ｎ (OCH) ２ CH ２ ) ｒ -、-K(CR ５ R ６ ) ｍ (O) (A) 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 (C) (A) ｔ (CR) ９ R １０ ) ｎ (OCH) ２ CH ２ ) ｒ -、-K(CR ５ R ６ ) ｍ (NR) １１ CON ｔ (CR) ９ R １０ ) ｎ (OCH) ２ CH ２ ) ｒ -、-K(CR ５ R ６ ) ｍ (O) (A) 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 (O.C.H. ２ CH ２ ) ｒ -, -K(CR ５ R ６ ) ｍ (CO) (Aa) ｔ (CR ９ R １０ ) ｎ (O.C.H. 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 ７ 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-methylpiperazine-(CO)(Aa) ｔ (CR ７ R ８ ) ｎ -, -K(CR ５ R) ｍ -(Aa) ｔ Phenyl-, -K(CR ５ R ６ ) ｍ -(Aa) ｔ Frill-, -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 where m and n are as defined above; Aa is a natural or unnatural amino acid; t and r are independently 0 to 100; R ３ , R ４ , R ５ , R ６ , R ７ , and R ８ are independently H; halide; C １ ~C ８ Alkyl; C ２ ~C ８ aryl, alkenyl, alkynyl, ether, ester, amine, or amide, and one or more of halide, CN, NR １ R ２ , C.F. ３ , OR １ , aryl, heterocycle, S(O)R １ , S.O. 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 selected from 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 (heterocycle or ３ ~C ８ (heteroaromatic ring), or a peptide containing 1 to 20 amino acids.

7. 2. The side-chain linked conjugate compound of formula (III) according to claim 1, or a pharma- ceutically acceptable salt, hydrate, or hydrate salt thereof, having any of the following structures: or a polymorphic crystal of these compounds; or an optical isomer, racemate, diastereomer, or enantiomer thereof: 【Chemistry 19】 【change】 【change】 【change】 During the ceremony, mAb is an antibody or cell binding molecule; X １ , X ２ , and X ３ are independently O, S, NH, NHNH, or CH ２ is; p, p １ , and p ２ are independently 1 to 100; q １ and q ２ is independently selected from 0 to 24; Aa is a natural or unnatural amino acid; r is 0-12; if r>2, (Aa)r is a peptide containing the same or different amino acid sequence; r=0 means that (Aa)r is absent; m and n are independently 1 to 30. R ２５ and R ２５’ is as defined in claim 3; Z ２ is as defined in claim 4; Ac is CH ３ It's COO-.

8. 3. The side chain linked compound of formula (V) of claim 2 having any of the following structures: 【Chemistry 21】 【change】 【change】 During the ceremony, X １ , X ２ , and X ３ are independently O, S, NH, NHNH, or CH ２ is; p, p １ , and p ２ are independently 1 to 100; q １ and q ２ is independently selected from 0 to 24; Aa is a natural or unnatural amino acid; r is 0-12; if r>2, (Aa)r is a peptide containing the same or different amino acid sequence; r=0 means that (Aa)r is absent; m is 1 to 30; R ２５ and R ２５’ is as defined in claim 3; Z ２ is as defined in claim 4; Lv ３ are 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; Ac is CH ３ COO-.

9. The cell binding agent / molecule is selected from the side-chain linked conjugate compounds of formula (III) according to claim 1 or 7: (A): antibodies, proteins, probodies, nanobodies, vitamins (including folates), peptides, polymer micelles, liposomes, lipoprotein-based drug carriers, nanoparticle drug carriers, dendrimers, and those bound to or coated with cell-binding ligands, or combinations of the above; (B); antibody-like proteins, full-length antibodies (polyclonal antibodies, monoclonal antibodies, antibody dimers, antibody multimers), multispecific antibodies (selected from bispecific antibodies, trispecific antibodies, or tetraspecific antibodies); single-chain antibodies, antibody fragments that bind to target cells, monoclonal antibodies, single-chain monoclonal antibodies, monoclonal antibody fragments that bind to target cells, chimeric antibodies, chimeric antibody fragments that bind to target cells, domain antibodies, domain antibody fragments that bind to target cells, resurfaced antibodies, single-chain resurfaced antibodies, and antibodies that bind to target cells. resurfaced antibodies, humanized antibodies, resurfaced humanized antibodies, single chain humanized antibodies, humanized antibody fragments that bind to target cells, anti-idiotypic (anti-Id) antibodies, CDR's, bispecifics, trispecifics, tetraspecifics, miniantibodies, probodies, probody fragments, small immune proteins (SIPs), lymphokines, hormones, vitamins, growth factors, colony stimulating factors, nutrient transport factors, large molecular weight proteins, fusion proteins, kinase inhibitors, gene targeting agents, nanoparticles, or polymers modified with antibodies or large molecular weight proteins; (C) Cell-binding ligands or receptor agonists selected from the following: folic acid derivatives; glutamic acid urea derivatives; somatostatin and its analogs (selected from the group consisting of octreotide (sandostatin) and lanreotide (somatorin); aromatic sulfonamides; pituitary adenylate cyclase activating peptide (PACAP) (PAC1); vasoactive intestinal peptide (VIP / PACAP) (VPAC1, VPAC2); melanocyte stimulating hormone (α-MSH); cholecystokinin (CCK) / gastrin receptor agonists; bombesin (Pyr-Gln-Arg-Leu-Gly-Asn-Gln-Trp-Ala-Val-Gly-His-Leu-Met-NH 2 Neurotensin receptor ligands (NTR1, NTR2, NTR3); substance P (NK1 receptor) ligands; neuropeptide Y (Y1-Y6); homing peptides including RGD (Arg-Gly-Asp), NGR (Asn-Gly-Arg), dimeric and multimeric cyclic RGD peptides (cRGDfVc), TAASGVRSMH and LTLRWVGLMS (chondroitin sulfate proteoglycan NG2 receptor ligand) and F3 peptide; cell penetrating peptides (CPPs); luteinizing homing peptides including RGD (Arg-Gly-Asp), NGR (Asn-Gly-Arg), dimeric and multimeric cyclic RGD peptides (cRGDfVc), TAASGVRSMH and LTLRWVGLMS (chondroitin sulfate proteoglycan NG2 receptor ligand) and F3 peptide; and buserelin (Pyr-His-Trp-Ser-Tyr-D-Ser(OtBu)-Leu-Arg-Pro-NHEt), gonadorelin (Pyr-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NHEt), which act by targeting follicle stimulating hormone (FSH) and luteinizing hormone (LH), as well as testosterone production, selected from the group consisting of lumen releasing hormone (LHRH) agonists and antagonists, and gonadotropin releasing hormone (GnRH) agonists. 2 ), goserelin (Pyr-His-Trp-Ser-Tyr-D-Ser(OtBu)-Leu-Arg-Pro-AzGly-NH2), histrelin (Pyr-His-Trp-Ser-Tyr-D-His(N-benzyl)-Leu-Arg-Pro-NHEt), leuprolide (Pyr-His-Trp-Ser-Tyr-D-Leu-Leu-Arg-Pro-NHEt), nafrelin (Pyr-His-Trp-Ser-Tyr-2Nal-Leu-Arg-Pro-Gly-NH 2 ), Triptorelin (Pyr-His-Trp-Ser-Tyr-D-Trp-Leu-Arg-Pro-Gly-NH 2 ), nafarelin, deslorelin, abarelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-(N-Me)Tyr-D-Asn-Leu-isopropyl-Pro-DAla-NH 2 ), Cetrorelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-Tyr-D-Cit-Leu-Arg-Pro-D-Ala-NH 2 ), Degarelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-4-amioPhe(L-hydroorotyl)-D-4-amioPhe(carbamoyl)-Leu-isopropylLys-Pro-D-Ala-NH 2 ), and ganirelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-Tyr-D-(N9, N10-diethyl)-homoArg-Leu-(N9, N10-diethyl)-homoArg-Pro-D-Ala-NH 2 ); pattern recognition receptors (PRRs) selected from the group consisting of Toll-like receptor (TLR) ligands, C-type lectins, and Nod-like receptor (NLRs) ligands; calcitonin receptor agonists; integrin receptors and their receptor subtypes (α Ｖ β １ , α Ｖ β ３ , α Ｖ β ５ , α Ｖ β ６ , α ６ β ４ , α ７ β １ , α Ｌ β ２ , α ＩＩｂ β ３ ) Agonists (GRGDSPK, cyclo(RGDfV)(L1) and its derivatives [cyclo(-N(Me)R-GDfV), cyclo(R-Sar-DfV), cyclo(RG-N(Me)D-fV), cyclo(RGD-N(Me)f-V), cyclo(RGDf-N(Me)V-) (ciliates)]; nanobodies (derivatives of VHH (camel Ig)); domain antibodies (dAb, derivatives of VH or VL domains), bispecific T cell induction (Bite, bispecific antibodies); dual affinity retargeting (DART, bispecific antibodies); tetravalent tandem antibodies (TandAb, dimerized bispecific antibodies); anticalins (derivatives of lipocalin); adnectins (tenth FN3 (fibronectin)); designed ankyrin repeat proteins (DARPins); avimers; EGF receptor and VEGF receptor agonists; (D): A small molecule of a cell-binding molecule / ligand or a cell receptor agonist selected from the following: LB01 (folic acid), LB02 (PMSA ligand), LB03 (PMSA ligand), LB04 (PMSA ligand), LB05 (somatostatin), LB06 (somatostatin), LB07 (octreotide, somatostatin analog), LB08 (ramareotide, somatostatin analog), LB09 (vapreotide (sambar), somatostatin analog), L B10 (CAIX ligand), LB11 (CAIX ligand), LB12 (gastrin releasing peptide receptor (GRPr), MBA), LB13 (luteinizing hormone releasing hormone (LH-RH) ligand and GnRH), LB14 (luteinizing hormone releasing hormone (LH-RH) and GnRH ligand), LB15 (GnRH antagonist, abarelix), LB16 (cobalamin, vitamin B12 analog), LB17 (cobalamin, vitamin B12 analog), LB18 (α ｖ β ３ for integrin receptors), LB19 (heterobivalent peptide ligand for VEGF receptors), LB20 (neuromedin B), LB21 (bombesin for G protein-coupled receptors), LB22 (TLR for Toll-like receptors), ２ ), LB23, LB24 (α ｖ Cilengitide / cyclo(-RGDfV-) for integrin receptors, LB25 (rifabutin analog), LB26 (rifabutin analog), LB27 (rifabutin analog), LB28 (fludrocortisone), LB29 (dexamethasone), LB30 (fluticasone propionate), LB31 (beclomethasone dipropionate), LB32 (triamcinolone acetonide), LB33 (prednisone), LB34 (prednisolone), LB35 (methylprednisolone), LB36 (betamethasone), LB37 (irinotecan analog), LB38 (crizotinib analog), LB 39 (bortezomib analog) carfilzomib analog), LB40 (carfilzomib analog), LB41 (carfilzomib analog), LB42 (leuprolide analog), LB43 (triptorelin analog), LB44 (clindamycin), LB45 (liraglutide analog), LB46 (semaglutide analog), LB47 (retapamulin analog), LB48 (indibulin analog), LB49 (vinblastine analog), LB50 (lixisenatide analog), LB51 (osimertinib analog), LB52 (nucleoside analog), LB53 (erlotinib analog), or LB54 (lapatinib analog); 【Chemical 22】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 During the ceremony, 【Chemistry 23】 is the site for linking to the side chain linker of the present invention; X ４ and Y １ are independently O, NH, NHNH, NR １ , S, C(O)O, C(O)NH, OC(O)NH, OC(O)O, NHC(O)NH, NHC(O)S, OC(O)N(R １ ), N(R １ ) C(O)N(R １ ), C.H. ２ , C(O)NHNHC(O), and C(O)NR １ is; １ is H, CH ２ , OH, O, C(O), C(O)NH, C(O)N(R １ ), R １ , N.H.R. １ , N.R. １ , C(O)R １ or C(O)O; X ５ is H, CH ３ , F, or Cl; １ and M. ２ are independently H, Na, K, Ca, Mg, NH ４ , N(R １ R ２ R ３ R ４ ) is; R １ , R ２ , R ３ , and R ４ is as defined in claim 1.

10. The side-chain linked conjugate compound of formula (III) according to claim 1 or 9, wherein the cell binding agent / molecule T has one or more of the following linkage structures: 【Chemistry 24】 In the formula, R ２０ and R ２１ is independent, C １ ~C ８ Alkyl; C ２ ~C ８ Heteroalkyl or heterocyclic ring; ３ ~C ８ Aryl, Ar-alkyl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, carbocycle, or alkylcarbonyl; or (CH ２ CH ２ O) ｐ C having the formula ２ ~C １００ It is polyethylene glycol.

11. The cell binding agent / molecule can target tumor cells, virus-infected cells, microorganism-infected cells, parasite-infected cells, autoimmune disease cells, activated tumor cells, bone marrow cells, activated T cells, affected B cells, melanocytes, or any cell expressing any one of the following antigens or receptors: CD1, CD1a, CD1b, CD1c, CD1d, CD1e, CD2, CD3, CD3d, CD3e, CD3g, CD4, CD5, CD6, CD7, CD8, CD8a, CD8b, CD9, CD1 0, CD11a, CD11b, CD11c, CD11d, CD12w, CD14, CD15, CD16, CD16a, CD16b, CDw 17, CD18, CD19, CD20, CD21, CD22, CD23, CD24, CD25, CD26, CD27, CD28, CD29 , CD30, CD31, CD32, CD32a, CD32b, CD33, CD34, CD35, CD36, CD37, CD38, CD39 , CD40, CD41, CD42, CD42a, CD42b, CD42c, CD42d, CD43, CD44, CD45, CD46, CD4 7, CD48, CD49b, CD49c, CD49c, CD49d, CD49f, CD50, CD51, CD52, CD53, CD54, CD55, CD56, CD57, CD58, CD59, CD60, CD60a, CD60b, CD60c, CD61, CD62E, CD6 2L, CD62P, CD63, CD64, CD65, CD65s, CD66, CD66a, CD66b, CD66c, CD66d, CD6 6e, CD66f, CD67, CD68, CD69, CD70, CD71, CD72, CD73, CD74, CD75, CD75s, CD7 6, CD77, CD78, CD79, CD79a, CD79b, CD80, CD81, CD82, CD83, CD84, CD85, CD8 5a, CD85b, CD85c, CD85d, CD85e, CD85f, CD85g, CD85g, CD85i, CD85j, CD85k , CD85m, CD86, CD87, CD88, CD89, CD90, CD91, CD92, CD93, CD94, CD95, CD96, CD97, CD98, CD99, CD100, CD101, CD102, CD103, CD104, CD105, CD106, CD107,CD107a、CD107b、CD108、CD109、CD110、CD111、CD112、CD113、CD114、CD115、CD116、CD117、CD118、CD119、CD120、CD120a、CD120b、CD121、CD121a、CD121b、CD122、CD123、CD123a、CD124、CD125、CD126、CD127、CD128、CD129、CD130、CD131、CD132、CD133、CD134、CD135、CD136、CD137、CD138、CD139、CD140、CD140a、CD140b、CD141、CD142、CD143、CD144、CD145、CDw145、CD146、CD147、CD148、CD149、CD150、CD151、CD152、CD153、CD154、CD155、CD156、CD156a、CD156b、CD156c、CD156d、CD157、CD158、CD158a、CD158b1、CD158b2、CD158c、CD158d、CD158e1、CD158e2、CD158f2、CD158g、CD158h、CD158i、CD158j、CD158k、CD159、CD159a、CD159b、CD159c、CD160、CD161、CD162、CD163、CD164、CD165、CD166、CD167、CD167a、CD167b、CD168、CD169、CD170、CD171、CD172、CD172a、CD172b、CD172g、CD173、CD174、CD175、CD175s、CD176、CD177、CD178、CD179、CD179a、CD179b、CD180、CD181、CD182、CD183、CD184、CD185、CD186、CDw186、CD187、CD188、CD189、CD190、CD191、CD192、CD193、CD194、CD195、CD196、CD197、CD198、CD199、CDw198、CDw199、CD200、CD201、CD202、CD202(a,b)、CD203、CD203c、CD204、CD205、CD206、CD207、CD208、CD209、CD210、CDw210a、CDw210b、CD211、CD212、CD213、CD213a1、CD213a2、CD214、CD215、CD216、<h2 style=";text-align:left;direction:ltr">CD2217、CD2218、CD2218a、CD2218、CD2218、CD221b9、CD2220、CD2221、CD22222、CD2223、CD2224、 CD225、CD226、CD227、CD228、CD229、CD230、CD231、CD232、CD233、CD234、CD2 35、CD235a、CD235b、CD236、CD237、CD238、CD239、CD240、CD240ce、CD240d、 CD241、CD242、CD243、CD244、CD245、C D246、CD247、CD248、CD249、CD250、CD2 51、CD252、CD253、CD254、CD255、CD256、CD257、CD258、CD259、CD260、CD261 、CD262、CD263、CD264、CD265、CD266、 CD267、CD268、CD269、CD270、CD271、CD 252、CD273、CD274、CD275、CD276、CD277、CD278、CD279、CD281、CD282、CD28 3、CD284、CD285、CD286、CD287、CD288 、CD289、CD290、CD291、CD292、CD293、C D294、CD295、CD296、CD297、CD298、CD 299、CD300、CD300a、CD300b、CD300c、 CD301、CD302、CD303、CD304、CD305、C D306、CD307、CD307a、CD307b、CD307c、 CD307d、CD307e、CD307f、CD308、CD30 9、CD310、CD311、CD312、CD313、CD314 、CD315、CD316、CD317、CD318、CD319、 CD320、CD321、CD322、CD323、CD324、CD 325、CD326、CD327、CD328、CD329、CD3 30、CD331、CD332、CD333、CD334、CD33 5、CD336、CD337、CD338、CD339、CD340 、CD341、CD342、CD343、CD344、CD345、C 5345、CD347、CD348、CD349、CD350、CD351、CD352、CD353、CD354、CD355、CD3 56、CD357、CD358、CD359、CD360、CD36 1、CD362、CD363、CD364、CD365、CD366、CD367, CD368, CD369, CD370, CD371, CD372, CD373, CD374, CD375, CD376, CD 377, CD378, CD379, CD381, CD382, CD383, CD384, CD385, CD386, CD387, CD388 , CD389, CRIPTO, CRIPTO, CR, CR1, CRGF, CRIPTO, CXCR5, LY64, TDGF1, 4-1BB , APO2, ASLG659, BMPR1B, 4-1BB, 5AC, 5T4 (chorionic glycoprotein, TPBG, 5T4, Wnt activation inhibitor antigen 1 or WAIF1), adenocarcinoma antigen, AGS-5, AGS-22M6, activin receptor-like kinase 1, AFP, AKAP-4, ALK, α integrin, αvβ6, aminopeptidase N, amyloid beta, androgen receptor, angiopoietin 2, angiopoietin 3, annexin A1, anthrax toxin protective antigen, anti-transferrin receptor, AOC3 (VAP-1), B7-H3, anthrax, BAFF (B-cell activating factor), BCMA, B-lymphoma cells, bcr-abl, bombesin, BORIS, C5, C242 antigen, CA125 (carbohydrate anti- Hara125, MUC16), CA-IX (or CAIX, carbonic anhydrase 9), CALLA, CanAg, canine IL31, carbonic anhydrase IX, cardiac myosin, CCL11 (C-C motif chemokine 11), CCR4 (CC chemokine receptor type 4), CCR5, CD3E (epsilon), CEA (carcinoembryonic antigen), CEACAM3, CEACAM5 (carcinoembryonic antigen), CFD (factor D), Ch4D5, cholecystokinin 2 (CCK2R), CLDN18 (claudin-18), clumping factor A, cMet, CRIPTO, FCSF1R (colony stimulating factor 1 receptor), CSF2 (colony stimulating factor 2, granulocyte-macrophage colony stimulating factor (GM-CSF)), CSP4, CTLA4 (cytotoxic T lymphocyte-associated protein 4), CTAA16.88 tumor antigen, CXCR4, CXC chemokine receptor type 4, cADP ribose hydrolase, cyclin B1, CYP1B1, cytomegalovirus, cytomegalovirus glycoprotein B, dabigatran, DLL4 (delta-like ligand 4), DPP4 (dipeptidyl peptidase 4), DR5 (death receptor 5), Escherichia coli Shiga toxin type 1, Escherichia coli Shiga toxin type 2,ED-B, EGFL7 (EGF-like domain-containing protein 7), EGFR, EGFRII, EGFRvIII, endoglin, endothelin B receptor, endotoxin, EpCAM (epithelial cell adhesion molecule), EphA2, episialin, ERBB2 (epidermal growth factor receptor 2), ERBB3, ERG (TMPRSS2ETS fusion gene), Escherichia coli, ETV6-AML, FAP (fibroblast activation protein protein α), FCGR1, α-fetoprotein, fibrin II, β chain, fibronectin ectodomain B, FOLR (folate receptor), folate receptor α, folate hydrolase, Fos-related antigen 1F, respiratory syncytial virus F protein, Frizzled receptor, fucosyl GM1, GD2 ganglioside, G-28 (cell surface glycolipid antigen), GD3 idiotype, GloboH, glypican 3, N-glucose Lycorylneuraminic acid, GM3, GMCSF receptor α chain, growth differentiation factor 8, GP100, GPNMB (transmembrane protein NMB), GUCY2C (guanylate cyclase 2C, guanylate cyclase C (GC-C), intestinal guanylate cyclase, guanylate cyclase-C receptor, heat-stable enterotoxin receptor (hSTAR)), heat shock protein, hemagglutinin, hepatitis B surface antigen, hepatitis B Hepatitis virus, HER1 (human epidermal growth factor receptor 1), HER2, HER2 / neu, HER3 (ERBB-3), IgG4, HGF / SF (stem cell growth factor / scatter factor), HHGFR, HIV-1, histone complex, HLA-DA (human leukocyte antigen), HLA-DR10, HLA-DRB, HMWMAA, human chorionic gonadotropin, HNGF, human scatter factor receptor kinase, HPV E6 / E7, Hsp90, hTERT, ICAM-1 (intercellular adhesion molecule 1), idiotype, IGF1R (IGF-1, insulin-like growth factor 1 receptor), IGHE, IFN-γ, influenza hemagglutinin, IgE, IgE Fc region, IGHE, interleukins (IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-6R, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-15, IL-17, IL-17A, IL-18, IL-19, IL-20, IL-21, IL-22, IL-23, IL-27, or IL-28), IL-31RA,ILGF2 (insulin-like growth factor 2), integrin (α4, α, ＩＩＩｂ β ３ , αvβ3, α ４ β ７ , α5β1, α6β4, α7β7, αIIβ3, α5β5, αvβ5), interferon-gamma-inducible protein, ITAGA2, ITGB2, KIR2D, LCK, Le , legumain, Lewis-Y antigen, LFA-1 (lymphocyte function-associated antigen 1, CD11a), LHRH, LINGO-1, lipoteichoic acid, LIV1A, LMP2, LTA, MAD-CT-1, MAD-CT-2, MAGE-1, MAGE-2, MAGE-3, MAGEA1, MAGEA3, MAGE4, ​​MART1, MCP-1, MIF (macrophage migration inhibitory factor or glycosylation inhibitory factor (GIF)), MS4A1 (member of transmembrane 4 domain subfamily A) 1), MSLN (mesothelin), MUC1 (mucin 1, cell surface associated (MUC1) or polymorphic epithelial mucin (PEM)), MUC1-KLH, MUC16 (CA125), MCP1 (monocyte chemotactic protein 1), MelanA / MART1, ML-IAP, MPG, MS4A1 (transmembrane 4 domain subfamily A), MYCN, myelin-associated glycoprotein, myostatin, NA17, NARP-1, NCA-90 (granulocyte antigen), nectin-4 (ASG-22M E), NGF, neuronal apoptosis-regulating proteinase 1, NOGO-A, Notch receptor, nucleolin, Neu oncogene product, NY-BR-1, NY-ESO-1, OX-40, OxLDL (oxidized low-density lipoprotein), OY-TES1, P21, p53 non-mutant, P97, Page4, PAP, anti-(N-glycolylneuraminic acid) paratope, PAX3, PAX5, PCSK9, PDCD1 (PD-1, programmed cell death protein 1), PDG F-Rα, (platelet-derived growth factor receptor α), PDGFR-β, PDL-1, PLAC1, PLAP-like testicular alkaline phosphatase, platelet-derived growth factor receptor β, sodium phosphate cotransporter, PMEL17, polysialic acid, proteinase 3 (PR1), prostate cancer, PS (phosphatidylserine), prostate cancer cells, Pseudomonas aeruginosa, PSMA, PSA, PSCA, rabies virus glycoprotein, RHD (Rh polypeptide 1 (RhPI)), rhesus factor (Rhesusfactor), RANKL, PhoC, Ras mutants, RG55, ROBO4, respiratory syncytial virus, RON, sarcoma metastasis breakpoint, SART3, sclerostin, SLAMF7 (SLAM family member 7), selectin P, SDC1 (syndecan 1), sLe(a), somatomedin C, SIP (sphingosine-1-phosphate), somatostatin, sperm protein 17, SSX2, STEA P1 (six-transmembrane epithelial antigen of the prostate 1), STEAP2, STn, TAG-22 (tumor-associated glycoprotein 72), survivin, T cell receptor, T cell transmembrane protein, TEM1 (tumor epithelial marker 1), TENB2, tenascin-C (TN-C), TGF-α, TGF-β (transforming growth factor β), TGF-β1, TGF-β2 (transforming growth factor β2), Tie (CD202b ), Tie2, TIM-1 (CDX-014), TN, TNF, TNF-α, TNFRSF8, TNFRSF10B (tumor necrosis factor receptor superfamily member 10B), TNFRSF13B (tumor necrosis factor receptor superfamily member 13B), TPBG (trophoblast glycoprotein), TRAIL-R1 (tumor necrosis-inducing ligand receptor 1), TRAILR2 (cell death receptor 5 (DR 5)), cells expressing major-associated calcium signal transducer 2, tumor-specific glycosylation of MUC1, TWEAK receptor, TYRP1 (glycoprotein 75), TRP-2, tyrosinase, VCAM-1, VEGF, VEGF-A, VEGF-2, VEGFR-1, VEGFR2, or vimentin, WT1, XAGE1, or any insulin growth factor receptor, or any epidermal growth factor receptor.

12. 12. The side chain linked conjugate compound of formula (III) of claim 11, wherein the tumor cells are selected from lymphoma cells, myeloma cells, renal cancer cells, breast cancer cells, prostate cancer cells, ovarian cancer cells, colon cancer cells, gastric cancer cells, squamous cell carcinoma cells, small cell lung cancer cells, non-small cell lung cancer cells, or testicular cancer cells.

13. 13. A pharmaceutical composition for the treatment or prevention of cancer, an autoimmune disease, or an infectious disease, comprising a therapeutically effective amount of a side-chain linked conjugate compound of formula (III) according to claim 1 or 7, and a pharma- ceutically acceptable salt, carrier, diluent, or excipient, or combination of the conjugate.

14. By weight, 0.01% to 99% of one or more side-chain-linked conjugate compounds of formula (III) according to claim 1 or 7; 0.0% to 20.0% of one or more polyols; 0.0% to 20.0% of one or more surfactants; 0.0% to 5.0% of one or more preservatives; 0.0% to 30.0% of one or more amino acids; 0.0% to 5.0% of one or more antioxidants; 0.0% to 0.3% of one or more chelating agents; 0.0% to 30.0% of one or more buffer salts to adjust the pH of the formulation to 4.5 to 7.5; 0.0% to 30.0% of one or more tonicity agents to adjust the osmolality to between 250-350 mOsm when reconstituted for administration to a patient; Including, the polyol is selected from fructose, mannose, maltose, lactose, arabinose, xylose, ribose, rhamnose, galactose, glucose, sucrose, trehalose, sorbose, melezitose, raffinose, mannitol, xylitol, erythritol, maltitol, lactitol, erythritol, threitol, sorbitol, glycerol, or L-gluconate or a metal salt thereof; The surfactants include polysorbate 20, polysorbate 40, polysorbate 65, polysorbate 80, polysorbate 81, or polysorbate 85, poloxamer, poly(ethylene oxide)-poly(propylene oxide), polyethylene-polypropylene, Triton; sodium dodecyl sulfate (SDS), sodium lauryl sulfate; sodium octyl glucoside; lauryl-, myristyl-, linoleyl-, or stearyl-sulfobetaine; lauryl-, myristyl-, linoleyl-, or stearyl-sarcosine; linoleyl-, myristyl-, or cetyl-betaine; lauroamidopropyl-, cocamidopropyl -, linoleamidopropyl-, myristamidopropyl-, palmidopropyl-, or isostearamidopropyl-betaine (lauroamidopropyl); myristamidopropyl-, palmidopropyl-, or isostearamidopropyl-dimethylamine; sodium methyl cocoyl taurate or disodium methyl oleyl taurate; dodecyl betaine, dodecyl dimethylamine oxide, cocamidopropyl betaine and cocoamphoglycinate; or isostearyl ethylimidonium ethosulfate; polyethyl glycol, polypropylene glycol, and copolymers of ethylene and propylene glycol; The preservative is selected from benzyl alcohol, octadecyldimethylbenzylammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl and benzyl alcohol, alkyl parabens, catechol, resorcinol, cyclohexanol, 3-pentanol, or m-cresol; The amino acids are selected from arginine, cystine, glycine, lysine, histidine, ornithine, isoleucine, leucine, alanine, glutamic acid or aspartic acid; The antioxidant is selected from ascorbic acid, glutathione, cystine, or methionine; The chelating agent is selected from EDTA or EGTA; The buffer salt is selected from sodium, potassium, ammonium, or trihydroxyethylamino salts of citric acid, ascorbic acid, gluconic acid, carbonic acid, tartaric acid, succinic acid, acetate, or phthalic acid; tris or tromethamine hydrochloride, phosphate, or sulfate; arginine, glycine, glycylglycine, or histidine and anionic acetate, chloride, phosphate, sulfate, or succinate; 14. The pharmaceutical composition of claim 13, wherein the isotonicity agent is selected from mannitol, sorbitol, sodium acetate, potassium chloride, sodium phosphate, potassium phosphate, trisodium citrate, or sodium chloride, either in a liquid formulation or in a formulated lyophilized solid.

15. 15. The pharmaceutical composition according to claim 13 or 14, which is filled in a vial, a bottle, a pre-filled syringe, or a pre-filled autoinjector syringe in liquid or lyophilized solid form.

16. A side-chain linked conjugate compound of formula (III) as claimed in claim 1 or 7, or a pharmaceutical composition as claimed in claim 13 or 14, having in vitro, in vivo or ex vivo cell killing activity.

17. 15. The pharmaceutical composition of claim 13 or 14, which is administered simultaneously with a chemotherapeutic agent, a radiotherapy, an immunotherapeutic agent, an autoimmune disease agent, an anti-infective agent, a synergist, or another conjugate for synergistically treating or preventing cancer, an autoimmune disease, or an infectious disease.

18. 18. The pharmaceutical composition of claim 17, wherein the chemotherapeutic agent is selected from the following: (1) a) Alkylating agents selected from the following: nitrogen mustards: chlorambucil, chlornaphazine, cyclophosphamide, dacarbazine, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, mannomustine, mitobronitol, melphalan, mitolactol, pipobroman, nobembitine, phenesterine, prednimustine, thiotepa, trofosfamide, uracil mustard; CC-1065 and synthetic analogs of adozelesin, carzelesin, and bizelesin; duocarmycin and its synthetic analogs KW-2189 and CBI-TMI; benzodiazepine dimers or pyrrolobenzodiazepine (PBD) dimers, tomaymycin dimers, indolinobenzodiazepine dimers. , imidazobenzothiadiazepine dimers, or oxazolidinobenzodiazepine dimers; nitrosourea compounds including carmustine, lomustine, chlorozotocin, fotemustine, nimustine, ranimustine; alkylsulfonates including busulfan, treosulfan, improsulfan, and piposulfan; triazenes or dacarbazine; platinum-containing compounds including carboplatin, cisplatin, and oxaliplatin; aziridines, benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramine, trimethylolomelamine; b) plant alkaloids selected from the group consisting of: vinca alkaloids including vincristine, vinblastine, vindesine, vinorelbine, navelbine; taxoids including paclitaxel and docetaxel and analogs thereof; maytansinoids including DM1, DM2, DM3, DM4, DM5, DM6, DM7, maytansine, ansamitocins and analogs thereof; cryptophycins including the cryptophycin 1 and cryptophycin 8 groups; epothilones, eleutherobin, discodermolide, bryostatins, dolostatins, auristatins, tubulysins, cephalostatins; pancratistatin; sarcodictiin; spongistatin; c) DNA topoisomerase inhibitors selected from: epipodophyllins, including 9-aminocamptothecin, camptothecin, crisnatol, daunomycin, etoposide, etoposide phosphate, irinotecan, mitoxantrone, novantrone, retinoic acid (retinols), teniposide, topotecan, 9-nitrocamptothecin or RFS 2000; and mitomycins and their analogs; d) Antimetabolites: {[Antifolates: (DHFR inhibitors: including methotrexate, trimetrexate, denopterin, pteropterin, aminopterin (4-aminopteroic acid), or other folate analogs); IMP dehydrogenase inhibitors (including mycophenolic acid, tiazofurin, ribavirin, EICAR); Ribonucleotide reductase inhibitors (including hydroxyurea, deferoxamine)]; [Pyrimidine analogs: uracil uridine analogs (including ancitabine, azacitidine, 6-azauridine, capecitabine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, 5-fluorouracil, floxuridine, raltitrexed); cytosine analogs: (including cytarabine, cytosine arabinoside, fludarabine); purine analogs: (including azathioprine, fludarabine, mercaptopurine, thiamiprine, thioguanine); folic acid supplements such as folinic acid}; e) Hormonal Therapy: Receptor Antagonists: [Antiestrogens: (including megestrol, raloxifene, tamoxifen); LHRH Agonists: (including goserelin, leuprolide acetate); Antiandrogens: (including bicalutamide, flutamide, calsterone, dromostanolone propionate, epithiostanol, goserelin, leuprolide, mepitiostane, nilutamide, testolactone, trilostane, and other similar androgen inhibitors. retinoids / deltoid: [vitamin D3 analogs: (including CB1093, EB1089, KH1060, cholecalciferol, ergocalciferol); photodynamic therapy agents: (including verteporfin, phthalocyanines, photosensitizer Pc4, demethoxyhypocrelin A); cytokines: (including interferon alpha, interferon gamma, tumor necrosis factor (TNF), TNF domain-containing human proteins)]}; f) Kinase inhibitors: selected from the group consisting of BIBW2992 (anti-EGFR / Erb2), imatinib, gefitinib, pegaptanib, sorafenib, dasatinib, sunitinib, erlotinib, nilotinib, lapatinib, axitinib, pazopanib, vandetanib, E7080 (anti-VEGFR2), mubritinib, ponatinib, bafetinib, bosutinib, cabozantinib, vismodegib, iniparib, ruxolitinib, CYT387, axitinib, tivozanib, sorafenib, bevacizumab, cetuximab, trastuzumab, ranibizumab, panitumumab, ispinesib; g) a poly(ADP-ribose)polymerase (PARP) inhibitor selected from the group consisting of olaparib, niraparib, iniparib, talazoparib, veliparib, CEP9722 (Cephalon), E7016 (Eisai), BGB-290 (Beigene), or 3-aminobenzamide; h) enediyne antibiotics (selected from the calicheamicins, calicheamicin gamma 1, delta 1, alpha 1, or beta 1; dynemicins including dynemicin A and deoxydynemicin; esperamicin, kedarcidin, C-1027, maduropeptin, or neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromophores), aclacinomycins, actinomycin, anthramycin, azaserine, bleomycins, cactinomycin, carabicin, carminomycin, carzinophilin; chromomycins, dactinomycin, daunomycin, Antibiotics selected from rubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin, epirubicin, eribulin, esorubicin, idarubicin, marcelomycin, nitomycins, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potofilomycin, puromycin, queramycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; i) Polyketides (acetogenins), bullatacin and bullatacinone; gemcitabine, epoxomicins and carfilzomib, bortezomib, thalidomide, lenalidomide, pomalidomide, tosedostat, zybrestat, PLX4032, STA-9090, Stimuvax, allovectin-7, zygeva, probenge, elvoy, isoprenylation inhibitors and lovastatin, dopaminergic neurotoxins and 1-methyl-4-phenyl Pyridine ion, cell cycle inhibitors (including staurosporine), actinomycins (including actinomycin D and dactinomycin), amanitins, bleomycins (including bleomycin A2, bleomycin B2, and peplomycin), anthracyclines (including daunorubicin, doxorubicin (adriamycin), idarubicin, epirubicin, pirarubicin, and zorubicin), mitoxantrone, MDR inhibitors or verapamil, Ca ２＋ ATP inhibitors or thapsigargin, histone deacetylase inhibitors (including vorinostat, romidepsin, panobinostat, valproic acid, mocetinostat (MGCD0103), belinostat, PCI-24781, entinostat, SB939, resminostat, gibinostat, AR-42, CUDC-101, sulforaphane, trichostatin A); thapsigargin, celecoxib, glitazones, epigallocatechin gallate, disulfiram, salinosporamide A, anti-adrenal drugs (selected from the group consisting of aminoglutethimide, mitotane, trilostanca); aceglatone; aldophosphamide glycoside; aminolevulinic acid; amsacrine; arabinoside, best Rabucil; Bisantrene; Edatrexate; Defofamine; Demecolcine; Diaziquone; Elfornithine (DFMO), Elfomitine; Elliptinium acetate; Etocruside, Gallium nitrate, Gacytosine, Hydroxyurea; Ibandronate, Lentinan; Lonidamine; Mitoguazone; Mopidamol; Nitracrine; Pentostatin; Fenamet; Pirarubicin; Podophyllic acid; 2-ethylhydrazide; Procarbazine; Razoxane; Rhizoxin; Sizofiran; Spirogermanium; Tenuazonic acid; Triaziquone; 2,2',2''-Trichlorotriethylamine; Trichothecenes (including T2 toxin, verrucarin A, roridin A, and anguidine); Urethane, siRNA, antisense medicines; 2) Anti-autoimmune disease drugs: cyclosporine, cyclosporine A, aminocaproic acid, azathioprine, bromocriptine, chlorambucil, chloroquine, cyclophosphamide, corticoids (including amcinonide, betamethasone, budesonide, hydrocortisone, flunisolide, fluticasone propionate, fluocortolone danazol, dexamethasone, triamcinolone acetonide, and beclomethasone dipropionate), DHEA, etanercept, hydroxychloroquine, infliximab, meloxicam, methotrexate, mofetil, mycophenolic acid, prednisone, sirolimus, and tacrolimus; 3) Anti-infective drugs: a) Aminoglycosides: amikacin, astromycin, gentamicin (netilmicin, sisomicin, isepamicin), hygromycin B, kanamycin (amikacin, arbekacin, bekanamycin, dibekacin, tobramycin), neomycin (framycin, paromomycin, ribostamycin), netilmicin, spectinomycin, streptomycin, tobramycin, verdamycin; b) Amphenicols: azidamphenicol, chloramphenicol, florfenicol, thiamphenicol; c) Ansamycins: geldanamycin, herbimycin; d) Carbapenems: biapenem, doripenem, ertapenem, imipenem, cilastatin, meropenem, panipenem; e) Cephalosporins: carbacephems (loracarbef), cephacetrile, cefaclor, cephradine, cefadroxil, cephalonium, cephaloridine, cephalothin or cephalothin, cephalexin, cephaloglycin, cefamandole, cephapirin, cefatrizine, cefazaflur, cefazedone, cefazolin, cefbuperazone, cefcapene, cefdaloxime, cefepime, cefminox, cefoxitin, cefprozil, cefroxadine, ceftezole, cefuroxime, cefixime, cefdinir, cefditoren, cefepime, Fetameth, cefmenoxime, cefodizime, cefonicid, cefoperazone, ceforanide, cefotaxime, cefotiam, cefozopran, cephalexin, cefpimizole, cefpiramide, cefpirome, cefpodoxime, cefprozil, cefquinome, cefsulodin, ceftazidime, cefteram, ceftibuten, ceftiolene, ceftizoxime, ceftobiprol, ceftriaxone, cefuroxime, cefuzonam, cephamycins (including cefoxitin, cefotetan, cefmetazole), oxacephems (flomoxef, latamoxef); f) Glycopeptides: bleomycin, vancomycin (including oritavancin, telavancin), teicoplanin (dalbavancin), ramoplanin; g) Glycylcyclines: Tigecycline; h) β-lactamase inhibitors: penams (sulbactam, tazobactam), clavams (clavulanic acid); i) Lincosamides: clindamycin, lincomycin; j) Lipopeptides: daptomycin, A54145, calcium-dependent antibiotic (CDA); k) Macrolides: azithromycin, cethromycin, clarithromycin, dirithromycin, erythromycin, flurithromycin, josamycin, ketolides (telithromycin, cethromycin), midecamycin, miokamicin, oleandomycin, rifamycin (rifampicin, rifampin, rifabutin, rifapentine), rokitamycin, roxithromycin, spectinomycin, spiramycin, tacrolimus (FK506), troleandomycin, telithromycin; l) Monobactams: aztreonam, tigemonam; m) Oxazolidinones: Linezolid; n) Penicillins: amoxicillin, ampicillin, pivampicillin, hetacillin, bacampicillin, methampicillin, talampicillin, azidocillin, azlocillin, benzylpenicillin, benzathine benzylpenicillin, benzathine phenoxymethylpenicillin, clometocillin, procaine benzylpenicillin, carbenicillin (carindacillin), cloxacillin, dicloxacillin, epicillin, flucloxacillin, mecillinam (pivmecillinam), mezlocillin, methicillin, nafcillin, oxacillin, penamecillin, penicillin, phenethicillin, phenoxymethylpenicillin, piperacillin, propicillin, sulbenicillin, temocillin, ticarcillin; o) Polypeptides: bacitracin, colistin, polymyxin B; p) quinolones: alatrofloxacin, balofloxacin, ciprofloxacin, clinafloxacin, danofloxacin, difloxacin, enoxacin, enrofloxacin, floxin, garenoxacin, gatifloxacin, gemifloxacin, grepafloxacin, canotrovafloxacin, levofloxacin, lomefloxacin, marbofloxacin, moxifloxacin, nadifloxacin, norfloxacin, orbifloxacin, ofloxacin, pefloxacin, trovafloxacin, grepafloxacin, sitafloxacin, sparfloxacin, temafloxacin, tosufloxacin, trovafloxacin; q) Streptogramins: pristinamycin, quinupristin / dalfopristin; r) Sulfonamides: mafenide, prontosil, sulfacetamide, sulfamethizole, sulfanilamide, sulfasalazine, sulfisoxazole, trimethoprim, trimethoprim-sulfamethoxazole (co-trimoxazole); s) steroidal antibacterial agents: selected from fusidic acid; t) Tetracyclines: doxycycline, chlortetracycline, clomocycline, demeclocycline, lymecycline, meclocycline, methacycline, minocycline, oxytetracycline, penimepicycline, rolitetracycline, tetracycline, glycylcyclines (including tigecycline); u) Other antibiotics: annonacin, arsphenamine, bactoprenol inhibitors (bacitracin), DADAL / AR inhibitors (cycloserine), dictyostatin, discodermolide, eleutherobin, epothilone, ethambutol, etoposide, faropenem, fusidic acid, furazolidone, isoniazid, laulimalide, metronidazole, mupirocin, mycolactone, NAM synthesis inhibitors (fosfomycin), nitrofurantoin, paclitaxel, platensimycin, pyrazinamide, quinupristin / dalfopristin, rifampicin (rifampin), tazobactamcinidazole, uvaricin; (4) Antiviral drugs: a) Entry / fusion inhibitors: aplaviroc, maraviroc, vicriviroc, gp41 (enfuvirtide), PRO140, CD4 (ibalizumab); b) Integrase inhibitors: raltegravir, elvitegravir, globoidnan A; c) Maturation inhibitors: Bevirimat, Vivicon; d) Neuraminidase inhibitors: oseltamivir, zanamivir, peramivir; e) Nucleosides and Nucleotides: Abacavir, Acyclovir, Adefovir, Amdoxovir, Apricitabine, Brivudine, Cidofovir, Clevudine, Dexerbucitabine, Didanosine (DDI), Elvucitabine, Emtricitabine (FTC), Entecavir, Famciclovir, Fluorouracil (5-FU), 3'-Fluoro substituted 2',3'-deoxynucleoside analogues (3'-Fluoro-2',3'-dideoxythymidine (FLT) and 3'-Fluoro-2',3'-di deoxyguanosine (FLG), fomivirsen, ganciclovir, idoxuridine, lamivudine (3TC), L-nucleosides (β-L-thymidine and β-L-2'-deoxycytidine), penciclovir, rasvir, ribavirin, stampidine, stavudine set (d4T), taribavirin (viramidine), telbivudine, tenofovir, trifluridine valacyclovir, valganciclovir, zalcitabine (ddC), zidovudine (AZT); f) Non-nucleosides: amantadine, ateviridine, capravirine, diarylpyrimidines (etravirine, rilpivirine), delavirdine, docosanol, emivirine, efavirenz, foscarnet (phosphorylformate), imiquimod, interferon alpha, loviride, rhodenosine, methisazone, nevirapine, NOV-205, peginterferon alpha, podophyllotoxin, rifampicin, rimantadine, resiquimod (R-848), tromantadine; g) Protease inhibitors: amprenavir, atazanavir, boceprevir, darunavir, fosamprenavir, indinavir, lopinavir, nelfinavir, pleconaril, ritonavir, saquinavir, telaprevir (VX-950), tipranavir; h) other types of antiviral drugs: abzymes, arbidol, calanolide a, ceragenin, cyanovirin-n, diarylpyrimidines, epigallocatechin gallate (EGCG), foscarnet, griffithsin, taribavirin (viramidine), hydroxycarbamide, KP-1461, miltefosine, pleconaril, portmanteau inhibitors, ribavirin, seliciclib; (5) A pharma-ceutically acceptable salt, acid, derivative, hydrate or hydrated salt; or crystal structure; or optical isomer, racemate, diastereomer or enantiomer of any of the above drugs.

19. The pharmaceutical composition of claim 17, wherein the synergist is selected from one or more of the following drugs: abatacept, abemaciclib, abiraterone acetate, abraxane, acetaminophen / hydrocodone, acalabrutinib, aducanumab, adalimumab, ADXS31-142, ADXS-HER2, afatinib dimalate, aldesleukin, alectinib, alemtuzumab, alitretinoin, ado-trastuzumab emtansine, amphetamine / dextroamphetamine, anastrozole, aripiprazole, anthracycline, aripiprazole, atazanavir, atezolizumab, atorvastatin, avelumab, axicabtagene. Ciloreucel, axitinib, belinostat, live BCG, bevacizumab, bexarotene, blinatumomab, bortezomib, bosutinib, brentuximab vedotin, brigatinib, budesonide, budesonide / formoterol, buprenorphine, cabazitaxel, cabozatinib, cammatinib, capecitabine, carfilzomib, chimeric antigen receptor-modified T (CAR-T) cells, celecoxib, ceritinib, cetuximab, chidamide, cyclosporine, cinacalcet, crizotinib, cobimetinib, cosentinib Tyx, crizotinib, CTL019, dabigatran, dabrafenib, dacarbazine, daclizumab, dacomitinib, daptomycin, daratumumab, darbepoetin alfa, darunavir, dasatinib, denileukin diftitox, denosumab, depakote, dexlansoprazole, dexmethylphenidate, dexamethasone, DigniCap Cooling System, dinutuximab, doxycycline, duloxetine, duvelisib, dalumabumab, elotuzumab, emtricibine / rilpivirine / tenofovir, disoproxil fumarate, emtricibine / tenofovir / efavirenz, enoxaparin, ensartinib, enzalutamide, epoetin alfa, erlotinib, esomeprazole, eszopiclone, etanercept, everolimus, exemestane, everolimus, exenatide ER, ezetimibe, ezetimibe / simvastatin, fenofibrate, filgrastim, fingolimod, fluticasone propionate, fluticasone / salmeterol, fulvestrant, gaziba,Gefitinib, Glatiramer, Goserelin Acetate, Icotinib, Imatinib, Ibritumomab Tiusetan, Ibrutinib, Idelalisib, Ifosfamide, Infliximab, Imiquimod, ImmuCyst, ImmunoBCG, Iniparib, Insulin Aspart, Insulin Detemir, Insulin Glargine, Insulin Lispro, Interferon Alpha, Interferon Alpha-1b, Interferon Alpha-2a, Interferon Alpha-2b, Interferon Beta, Interferon Beta-1a, Interferon Beta-1b, Insulin Interferon gamma-1a, lapatinib, ipilimumab, ipratropium bromide / salbutamol, ixazomib, Kanuma, lanreotide acetate, lenalidomide, lenaliomide, lenvatinib mesylate, letrozole, levothyroxine, lidocaine, linezolid, liraglutide, lisdexamfetamine, LN-144, loratinib, memantine, methylphenidate, metoprolol, mekinist, mericitabine / rilpivirine / tenofovir, modafinil, mometasone, Mycidac-C, necitumumab, Neratinib, nilotinib, niraparib, nivolumab, ofatumumab, obinutuzumab, olaparib, olmesartan, olmesartan / hydrochlorothiazide, omalizumab, omega-3 fatty acid ethyl ester, oncoline, oseltamivir, osimertinib, oxycodone, palbociclib, palivizumab, panitumumab, panobinostat, pazopanib, pembrolizumab, PD-1 antibody, PD-L1 antibody, pemetrexed, pertuzumab, pneumococcal conjugate vaccine, pomalidomide, pregabalin, ProscaVax, propranolol, Nolol, quetiapine, rabeprazole, radium 223 chloride, raloxifene, raltegravir, ramucirumab, ranibizumab, regorafenib, ribociclib, rituximab, rivaroxaban, romidepsin, rosuvastatin, ruxolitinib phosphate, salbutamol, savolitinib, semaglutide, sevelamer, sildenafil, siltuximab, sipuleusel-T, sitagliptin, sitagliptin / metformin, solifenacin, solanezumab, sonidegib, sorafenib, sunitinib, tacrolimus,Taclimus, Tadalafil, Tamoxifen, Tafinlar, Talimogenelaherparepvec, Talazoparib, Telaprevir, Talazoparib, Temozolomide, Tensirolimus, Tenofovir / Emtricitabine, Tenofovir Disoproxil Fumarate, Testosterone Gel, Thalidomide, TICE BCG, Tiotropium Bromide, Tisagenlecleucel, Toremifene, Trametinib, Trastuzumab, Trabectin (ecteinascidin 743), Trametinib, Tremelimumab, Trifluridine / Tipiracil, Tretinoin, Uro-BCG, Ustekinumab, Valsartan, Veliparib, Vandetanib, Vemurafenib, Venetoclax, Vorinostat, Dib-Aflibercept, and Zostavax, as well as analogs, derivatives, pharma- ceutically acceptable salts, carriers, diluents, or excipients, or combinations thereof.

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