# US 2018/0021448 A9 · Conjugates of Cell Binding Molecules with Cytotoxic Agents

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- Number: US 2018/0021448 A9
- Document ID: US20180021448A9
- Status: Published application
- Kind: A9
- Application number: 14/253,881
- Filed: 2014-04-16
- Published: 2018-01-25
- Earliest claimed: 2012-07-12
- Assignees: Hangzhou Dac Biotech Co., Ltd.
- Applicants: Hangzhou DAC Biotech Co., Ltd.
- Inventors: R. Yongxin Zhao; Yue Zhang; Yourang Ma
- Web page: https://patentgrep.com/patent/US20180021448A9

## CPC

- C07D 207/40: 2,5-Pyrrolidine-diones
- A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
- A61K 47/64: Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent (peptidic linkers A61K47/65)
- A61K 47/6803: {Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates}
- A61K 47/6811: {the drug being a protein or peptide, e.g. transferrin or bleomycin}
- A61K 47/6849: {the antibody targeting a receptor, a cell surface antigen or a cell surface determinant}
- A61K 47/6851: {the antibody targeting a determinant of a tumour cell}
- A61K 47/6855: {the tumour determinant being from breast cancer cell}
- A61K 47/6863: {the tumour determinant being from stomach or intestines cancer cell}
- A61K 47/6867: {the tumour determinant being from a cell of a blood cancer}
- A61P 31/00: Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P 31/04: Antibacterial agents
- A61P 35/00: Antineoplastic agents
- A61P 37/00: Drugs for immunological or allergic disorders
- A61P 37/02: Immunomodulators
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
- C07D: HETEROCYCLIC COMPOUNDS (macromolecular compounds C08)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

A conjugate of a potent cytotoxic agent with a cell-surface receptor binding molecule having a Formula (I), wherein T, L, m, n, R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12, and R13 are defined herein, can be used for targeted treatment of cancer, autoimmune disease, and infectious disease.

## Claims

### Claim 1

A conjugate of a cell binding molecule with cytotoxic agents having a structure of Formula (I):
and pharmaceutical acceptable salts and solvates thereof,
Wherein T is a targeting or binding ligand; L is a releasable linker; ----- is a linkage bond that L connects to an atom inside the bracket independently; n is 1˜20 and m is 1˜10.
Inside the bracket is a potent antimitotic agent wherein R1, R2, R3, and R4 are independently C1˜C8 of alkyl, heteroalkyl; C2˜C8 of heterocyclic, carbocyclic, cycloalkyl, alkylcycloalkyl, heterocycloalkyl; C3˜C8 of aryl, Ar-alkyl, heteroalkylcycloalkyl, alkylcarbonyl; or two R's, such as R1R2, R1R3, R2 R3, R3 R4, R5 R6, and R12 R13 can be 3˜7 members of a carbocyclic, cycloalkyl, or heterocyclic, heterocycloalkyl ring system; Y is N or CH; In addition, R1, R3, and R4 can be H, and R2 can be absent.
Wherein R5, R6, R8 and R10 are independently selected from H and C1˜C4 of alkyl or heteroalkyl.
Wherein R7 is independently selected from H, R14, or —R14C(═O)X1R15; —R14X1R15, wherein R14 and R15 are independently selected from C1˜C8 of alkyl, heteroalkyl; C2˜C8 of alkenyl, alkynyl, heterocyclic, carbocyclic, cycloalkyl; C3˜C8 of aryl, heterocycloalkyl, heteroalkylcycloalkyl, heteroaralkyl, alkylcarbonyl; X1 is selected from O, S, S—S, NH, or NR14.
Wherein R9 is independently H, —O—, —OR14, —OC(═O)R14—, —OC(═O)NHR14—, —OC(═O) R14SSR15—, OP(═O)(OR14)—, or OR14OP(═O)(OR15), wherein R14 and R15 are independently C1˜C8 of alkyl, heteroalkyl; C2˜C8 of alkenyl, alkynyl, heterocyclic, carbocyclic; C3˜C8 of aryl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, heteroaralkyl, alkylcarbonyl.
Wherein R11 is independently H, R14, —R14C(═O)R16, —R14X2R16, —R14C(═O)X2, wherein X2 is —O—, —S—, —NH—, —N(R14)—, —O—R14—, —S—R14—, —S(═O)—R14—, or —NHR14—; R14 is C1˜C8 of alkyl, heteroalkyl; C2˜C8 of alkenyl, alkynyl, heterocyclic, carbocyclic; C3˜C8 of aryl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, heteroaralkyl, alkylcarbonyl; R16 is H, OH, R14, or one to four amino acid units;
Wherein R12 is independently R14, —O—, —S—, —N—, ═N—, ═NNH—, —OH, —SH, —NH2, ═NH, ═NNH2, —NH(R14), —OR14, —C(O)O—, —C(O)OR16—, —COR16, —COOR14—, C(O)NH—, C(O)NH2, C(O)NHR14, —SR14, —S(═O)R14, —P(═O)(OR16)2, —OP(═O)(OR16)2, —CH2OP(═O)(OR16)2, —SO2R16. R14 is independently C1˜C8 of alkyl, heteroalkyl; C2˜C8 of alkenyl, alkynyl, hetero-cyclic, carbocyclic; C3˜C8 of aryl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroaralkyl, heteroalkylcycloalkyl, alkylcarbonyl. R16 is H, OH, R14 or one to four amino acid units;
Wherein R13 is C1˜C10 of alkyl, heteroalkyl, alkyl acid, alkyl amide, alkyl amine, or Ar; Ar refers to an aromatic or hetero aromatic group, composed of one or several rings, comprising four to ten carbon atoms, preferentially four to six carbon atoms. The term of hetero aromatic group refers to an aromatic group that has one or several carbon atoms replaced by hetero atoms, preferentially one, two or three carbon atoms replaced by O, N, Si, Se, P or S, more preferentially O, S, N. The term aryl or Ar also refers to an aromatic group, wherein one or several H atoms can be replaced independently by R17, F, Cl, Br, I, OR16, SR16, NR16R17, N═NR16, N═R16, NR16 R17, NO2, SOR16R17, SO2 R16, SO3 R16, OSO3 R16, PR16R17, POR16R17, PO2R16R17, P(O)(OR17)2, OCH2OP(O)(OR17)2, OC(O)OP(O)(OR17)2, PO(OR16)(OR17), OP(O)(OR17)OP(O)(OR17)2, OC(O)R17 or OC(O)NHR17, wherein R16, R17 are independently H, C1˜C8 of alkyl, heteroalkyl; C2˜C8 of alkenyl, alkynyl, heterocyclic, carbocyclic; C3˜C8 of aryl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, heteroaralkyl, alkylcarbonyl or C4˜C12 of glycosides, or pharmaceutical salts.
In addition, R12 can be H when R10 is not H, or when R13 is:
wherein Z1 is H, CH2OP(O)(OR18)2, C(O)OP(O)(OR18)2, PO(OR18)2, P(O)(OR18)OP(O)(OR18)2, C(O)R18, C(O)NHR18, SO2(OR18), C4˜C12 glycosides, or C1˜C8 of alkyl, carboxyalkyl, heterocyclic; R18 is H, C1˜C8 of alkyl, carboxyalkyl, heteroalkyl; C2˜C8 of alkenyl, alkynyl, heterocyclic; C3˜C8 of aryl, alkylcarbonyl; R19 is H, OH, NH2, OSO2(OR18), XCH2OP(O)(OR18)2, XPO(OR18)2, XC(O)OP(O)(OR18)2, XC(O) R18, XC(O)NHR18, C1˜C8 of alkyl, carboxyalkyl, carboxylic acid derivative; C2˜C8 of alkenyl, alkynyl, heterocyclic; C3˜C8 of aryl, alkylcarbonyl; or pharmaceutical salts; X is O, S, NH; Y1 and Y2 are N or CH respectively.
Or R12 can be H when R11 is:
wherein X2 is O, S, N—R8; R6 and R8 are H, C1˜C6 of alkyl or heteroalkyl.
Wherein the said conjugate of the invention specifically excludes the following structures:
Wherein R24 is H or CH3; R25 is alkyl or C(O)alkyl or C(O)aryl; R26 is H, C1˜C5 alkyl, C2˜C5 alkenyl, C(═O)C1˜C5 alkyl, or C(═O)C2˜C5 alkenyl; R28 is H or C1˜C5 alkyl; R27 is H, 1 to 3 substituents selected from halo, nitro, carboxylate or a derivative thereof, cyano, hydroxyl, alkyl, haloalkyl, alkoxy, haloalkoxy, phenol protecting groups, prodrug moieties, or OR29, where R29 is substituted aryl, C(O)R30, P(O)(OR30)2, or SO3R30, where R30 is selected from H, alkyl, alkenyl, cycloalkyl, heterocyclyl, aryl, and arylalkyl or a metal cation; T is a targeting or binding ligand; L is a releasable linker.

### Claim 2 (depends on 1)

A conjugate of cell binding molecule-cytotoxic agent according to claim 1 having Formula (II):
and pharmaceutical acceptable salts and solvates thereof,
Wherein T is a targeting or binding ligand; L is a releasable linker that connects to a molecule inside the bracket independently; n is 1˜20 and m is 1˜10.
Inside the bracket is a potent antimitotic agent wherein R1, R2, R3, and R4 are independently C1˜C8 of alkyl, heteroalkyl; C2˜C8 of heterocyclic, carbocyclic, cycloalkyl, alkylcycloalkyl, heterocycloalkyl; C3˜C8 of aryl, Ar-alkyl, heteroalkylcycloalkyl, alkylcarbonyl; or two R's, such as R1R2, R1R3, R2 R3, R3 R4, R5 R6, and R12 R13 can be 3˜7 members of a carbocyclic, cycloalkyl, or heterocyclic, heterocycloalkyl ring system; is N or CH; In addition, R1, R3, and R4 can be H, and R2 can be absent.
Wherein R5, R6, R8 and R10 are independently selected from H and C1˜C4 of alkyl or heteroalkyl.
Wherein R7 is independently selected from H, R14, or —R14C(═O)X1R15; —R14X1R15, wherein R14 and R15 are independently selected from H, C1˜C8 of alkyl, heteroalkyl; C2˜C8 of alkenyl, alkynyl, heterocyclic, carbocyclic, cycloalkyl; C3˜C8 of aryl, heterocycloalkyl, heteroalkylcycloalkyl, heteroaralkyl, alkylcarbonyl; X1 is selected from O, S, S—S, NH, or NR14.
Wherein R9 is independently H, —O—, —OR14, —OC(═O)R14—, —OC(═O)NHR14—, —OC(═O) R14SSR15—, OP(═O)(OR14)—, or OR14OP(═O)(OR15), wherein R14, R15 are independently H, C1˜C8 of alkyl, heteroalkyl; C2˜C8 of alkenyl, alkynyl, heterocyclic, carbocyclic; C3˜C8 of aryl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, heteroaralkyl, alkylcarbonyl.
Wherein R11 is independently H, R14, —R14C(═O)R16, —R14X2R16, —R14C(═O)X2, wherein X2 is —O—, —S—, —NH—, —N(R14)—, —O—R14—, —S—R14—, —S(═O)—R14—, or —NHR14—; R14 is H, C1˜C8 of alkyl, heteroalkyl; C2˜C8 of alkenyl, alkynyl, heterocyclic, carbocyclic; C3˜C8 of aryl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, heteroaralkyl, alkylcarbonyl; R16 is H, OH, R14, or one to four amino acid units;
Wherein R12 is independently R14, —OH, —SH, —NH2, ═NH, ═NNH2, —NH(R14), —OR14, —COR16, —COOR14—, C(O)NH2, C(O)NHR14, —SR14, —S(═O)R14, —P(═O)(OR16)2, —OP(═O)(OR16)2, —CH2OP(═O)(OR16)2, —SO2R16. R14 is independently H, C1˜C8 of alkyl, heteroalkyl; C2˜C8 of alkenyl, alkynyl, hetero-cyclic, carbocyclic; C3˜C8 of aryl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroaralkyl, heteroalkylcycloalkyl, alkylcarbonyl. R16 is H, OH, R14 or one to four amino acid units;
Wherein R13 is C1˜C10 of alkyl, heteroalkyl, alkyl acid, alkyl amide, alkyl amine, or Ar; Ar refers to an aromatic or hetero aromatic group, composed of one or several rings, comprising four to ten carbon atoms, preferentially four to six carbon atoms. The term of hetero aromatic group refers to an aromatic group that has one or several carbon atoms replaced by hetero atoms, preferentially one, two or three carbon atoms replaced by O, N, Si, Se, P or S, preferentially O, S, N. The term aryl or Ar also refers to an aromatic group, wherein one or several H atoms are replaced independently by R17, F, Cl, Br, I, OR16, SR16 NR16R17, N═NR16, N═R16, NR16 R17, NO2 SOR16R17, SO2 R16, SO3 R16, OSO3 R16, PR16R17 POR16R17, PO2R16R17, OP(O)(OR17)2, OCH2OP(O)(OR17)2, OC(O)OP(O)(OR17)2, PO(OR16)(OR17), OP(O)(OR17)OP(O)(OR17)2, OC(O)R17 or OC(O)NHR17, wherein R16, R17 are independently H, C1˜C8 of alkyl, heteroalkyl; C2˜C8 of alkenyl, alkynyl, heterocyclic, carbocyclic; C3˜C8 of aryl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, heteroaralkyl, alkylcarbonyl or C4˜C12 of glycosides, or pharmaceutical salts.
In addition, R12 can be H when R1 is not H, or when R13 is:
wherein Z1 is H, CH2OP(O)(OR18)2, C(O)OP(O)(OR18)2, PO(OR18)2, P(O)(OR18)OP(O(O)(OR18)2, C(O)R18, C(O)NHR18, SO2(OR18), C4˜C12 glycosides or C1˜C8 of alkyl, carboxyalkyl, heterocyclic; R18 is H, C1˜C8 of alkyl, carboxyalkyl, heteroalkyl; C2˜C8 of alkenyl, alkynyl, heterocyclic; C3˜C8 of aryl, alkylcarbonyl; R19 is H, OH, NH2, OSO2(OR18), XCH2OP(O)(OR18)2, XPO(OR18)2, XC(O)OP(O)(OR18)2, XC(O) R18, XC(O)NHR18, C1˜C8 of alkyl, carboxyalkyl, carboxylic acid derivative; C2˜C8 of alkenyl, alkynyl, heterocyclic; C3˜C8 of aryl, alkylcarbonyl; or pharmaceutical salts; X is O, S, NH; Y1 and Y2 are N or CH respectively.
Or R12 can be H when R11 is:
wherein X2 is O, S, N—R5; R6 and R8 are H, C1˜C6 of alkyl or heteroalkyl.

### Claim 3 (depends on 1, 2)

A cell binding ligand-antimitotic agent conjugate according to claim 1 having the structure of Formula (III):
and pharmaceutical acceptable salts and solvates thereof,
Wherein T, L, n, m, Y, R1, R2, R3, R4, R5, R6, R8, R10, R11 and R13 are defined the same as in the claim 1.
Wherein R9 and R12 are defined the same as in the claim 2.
Wherein R7 is independently selected from R14 or —R14C(═O)X1R15—, or —R14X1R15—; wherein R14 and R15 are independently selected from H, C1˜C8 of alkyl, heteroalkyl; C2˜C8 of alkenyl, alkynyl, heterocyclic, carbocyclic, cycloalkyl; C3˜C8 of aryl, heterocycloalkyl, heteroalkylcycloalkyl, heteroaralkyl, alkylcarbonyl; X1 is selected from O, S, S—S, NH, or NR14.
In addition, R7 can be absent.

### Claim 4 (depends on 1, 2)

The conjugate according to claim 1, having the structure of Formula (IV):
and pharmaceutical acceptable salts and solvates thereof,
Wherein T, L, n, m, Y, R1, R2, R3, R4, R5, R6, R7, R8, R10, R11 and R13 are defined the same as in the claim 1.
Wherein R12 is defined the same as in the claim 2.
Wherein R9 is independently —O—, —OR14—, —OC(═O)R14—, —OC(═O)NHR14—, —OC(═O) R14SSR15—, OP(═O)(OR14)O—, wherein R14, R5 are independently H C1˜C8 of alkyl, heteroalkyl; C3˜C8 of aryl, heteroaryl, heterocyclic, carbocyclic, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, heteroaralkyl, alkylcarbonyl or pharmaceutical salts. In addition, R9 can be absent.

### Claim 5 (depends on 1, 2)

A conjugate of cell binding molecule-cytotoxic agent according to claim 1 having a structure of Formula (V):
and pharmaceutical acceptable salts and solvates thereof,
Wherein T, L, n, m, Y, R1, R2, R3, R4, R5, R6, R7, R8, R10 and R13 are defined the same as in the claim 1.
Wherein R9 and R12 are defined the same as in the claim 2.
Wherein R11 is independently —R14—, —R14C(═O)R17—, —R14X2R17—, —R14C(═O)X2—, wherein R17 is independently H, OH, C1˜C8 of alkyl; C2˜C8 of alkenyl, alkynyl, heteroalkyl; C3˜C8 of aryl, arylene, heterocyclic, carbocyclic, heterocycloalkyl, or an amino acid, or two amino acid; X2 is —O—, —S—, —NH—, —N(R14)—, —O—R14—, —S—R14—, —S(═O)—R14—, or —NHR14—; R14 is C1˜C8 of alkyl, heteroalkyl; C2˜C8 of alkenyl, alkynyl, heterocyclic, carbocyclic; C3˜C8 of aryl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, heteroaralkyl, alkylcarbonyl.
Wherein said conjugate of the formula (V) specifically excludes the following structures:
Wherein R25 is alkyl or C(O)alkyl or C(O)aryl; R26 is H, C1˜C5 alkyl, C2˜C5 alkenyl, C(═O)C1˜C5 alkyl, or C(═O)C2˜C5 alkenyl; R27 is H, 1 to 3 substituents selected from halo, nitro, carboxylate or a derivative thereof, cyno, hydroxyl, alkyl, haloalkyl, alkoxy, haloalkoxy, phenol protecting groups, prodrug moieties, and OR29, where R29 is substituted aryl, C(O)R30, P(O)(OR30)2, or SO2R30, where R30 is selected from H, alkyl, alkenyl, cycloalkyl, heterocyclyl, aryl, and arylalkyl or a metal cation; T is a targeting or binding ligand; L is a releasable linker.

### Claim 6 (depends on 1, 2)

A conjugate of cell binding molecule-cytotoxic agent according to claim 1 having a structure of Formula (VI):
and pharmaceutical acceptable salts and solvates thereof,
Wherein T, L, n, m, R1, R2, R3, R4, R5, R6, R7, R8, R10, R11 and R13 are defined the same as in the claim 1.
Wherein R9 is defined the same as in the claim 2.
Wherein R12 is independently R14, —O—, —S—, —NH—, ═N—, ═NNH—, —N(R14)—, —OR14—, —C(O)O—, —C(O)NH—, C(O)NR14—, —SR14—, —S(═O)R14—, —NHR14—, —CH2OP(═O)(OR15)—, —P(═O)(OR15)—, —C(O)OP(═O)(OR15)—, —OP(═O)(OR15)O−, —SO2R14. R14 and R15 are independently H, C1˜C8 of alkyl, heteroalkyl; C2˜C8 of alkenyl, alkynyl, heterocyclic, carbocyclic; C3˜C8 of aryl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroaralkyl, heteroalkylcycloalkyl, alkylcarbonyl.
In addition, R12 can be H when R10 is not H, or when R13 is:
wherein Z1 is H, CH2OP(O)(OR18)2, C(O)OP(O)(OR18)2, PO(OR18)2, P(O)(R18)OP(O)(OR18)2, C(O)R18, C(O)NHR18, SO2(OR18), C4˜C12 glycosides or C1˜C8 of alkyl, carboxyalkyl, heterocyclic; R18 is H, C1˜C8 of alkyl, carboxyalkyl, heteroalkyl; C2˜C8 of alkenyl, alkynyl, heterocyclic; C3˜C8 of aryl, alkylcarbonyl; R19 is H, OH, NH2, OSO2(OR18), XCH2OP(O)(OR18)2, XPO(OR18)2, XC(O)OP(O)(R18)2, XC(O) R18, XC(O)NHR18, C1˜C8 of alkyl, carboxyalkyl, carboxylic acid derivative; C2˜C8 of alkenyl, alkynyl, heterocyclic; C3˜C8 of aryl, alkylcarbonyl; or pharmaceutical salts; X is O, S, NH; Y1 and Y2 are N or CH respectively.
Or R12 can be H when R11 is:
wherein X2 is O, S, N—R8; R6 and R8 are H, C1˜C6 of alkyl or heteroalkyl respectively.

### Claim 7 (depends on 1, 2)

A conjugate of cell binding molecule-cytotoxic agent according to claim 1 having a structure of Formula (VII):
and pharmaceutical acceptable salts and solvates thereof,
Wherein T, L, n, m, Y, R1, R2, R3, R4, R5, R6, R7, R8, R10, R11 and R13 are defined the same as in the claim 1.
Wherein R9 and R12 are defined the same as in the claim 2.
Wherein R13 is C1˜C10 of alkyl, heteroalkyl, alkyl acid, alkyl amide, alkyl amine, or Ar; Ar refers to an aromatic or hetero aromatic group, composed of one or several rings, comprising four to ten carbon atoms, preferentially four to six carbon atoms. The term of hetero aromatic group refers to an aromatic group that has one or several carbon atoms replaced by hetero atoms, preferentially one, two or three carbon atoms replaced by O, N, Si, Se, P or S, preferentially O, S, N. The term aryl or Ar also refers to an aromatic group, wherein one or several H atoms are replaced independently by R18, F, Cl, Br, I, OR16, SR16, NR16R17, N═NR16, N═R16, NR16 R18, NO2, SOR16R18, SO2R16, SO3R16, OSO3R16, PR16R18, POR16R18, PO2R16R18, PO3OR16R18, CH2OPO3R16R18, OP(O)(OR16)OP(O)(OR18)2, C(O)OPO3R16R18, or PO3R16R18, wherein R16, R18 are independently H, C1˜C8 of alkyl; C2˜C8 of alkenyl, alkynyl, heteroalkyl; C3˜C8 of aryl, heterocyclic, carbocyclic, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroaralkyl, heteroalkylcycloalkyl, alkylcarbonyl; or C4˜C12 of glycosides; or pharmaceutical salts.
Wherein said conjugate of the formula (VII) specifically excludes the following structures:
Wherein R24 is H or CH3; R25 is alkyl or C(O)alkyl or C(O)aryl; R26 is H, C1˜C5 alkyl, C2˜C5 alkenyl, C(═O)C1˜C5 alkyl, or C(═O)C2˜C5 alkenyl; R28 is H or C1˜C5 alkyl; T is a targeting or binding ligand; L is a releasable linker.

### Claim 8 (depends on 1)

The conjugate according to claim 1, having the structure of Formula (VIII):
and pharmaceutical acceptable salts and solvates thereof,
Wherein T is a targeting or binding ligand; L is a releasable linker that connects to a molecule inside the bracket independently; n is 1˜20 and m is 1˜10; q=1˜5;
Wherein Z1 is H, CH2OP(O)(OR18)2, PO(OR18)2, C(O)OP(O)(OR18)2, P(O)(OR18)OP(O)(OR18)2, C(O)R18, C(O)NHR18, SO2(OR18), C4˜C12 of glycosides, or C1˜C8 of alkyl, carboxyalkyl, heterocyclic; R18 is H, C1˜C8 of alkyl, carboxyalkyl, heteroalkyl; C2˜C8 of alkenyl, alkynyl, heterocyclic; C3˜C8 of aryl, alkylcarbonyl; R24 and R24′ are independently H or CH3.

### Claim 9 (depends on 1, 2, 3, 4, 5, 6, 7, 8)

The conjugate of claims 1, 2, 3, 4, 5, 6, 7 and 8 wherein the linker L has the formula of: —Ww-(Aa)r-Vv—
Wherein: —W— is a stretcher unit, which links a targeted binding molecular unit (T) to an amino acid unit (Aa) when presents, or links V when an Aa is not present, and W may independently contain a self-immolative spacer, a peptidyl unit, a hydrazone, a disulfide, a thioether bond, an ester or an amide bond; w is 0 or 1; Wherein Aa is independently a natural or unnatural amino acid unit; r is independently an integer ranging from 0 to 12; (Aa)r represents a natural or unnatural amino acid, dipeptide, tripeptide, tetrapeptide, pentapeptide, hexapeptide, heptapeptide, octapeptide, nonapeptide, decapeptide, undecapeptide or dodecapeptide unit.
Wherein V is a spacer unit and is independently H, O, NH, S, C1˜C8 of alkyl, heteroalkyl; C2˜C8 of alkenyl, alkynyl, hetero-cyclic, carbocyclic; C3˜C8 of aryl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroaralkyl, heteroalkylcycloalkyl, alkylcarbonyl, or one to four amino acid units; and v is 0, 1 or 2.

### Claim 10 (depends on 1, 2, 3, 4, 5, 6, 7, 8)

The conjugate of claims 1, 2, 3, 4, 5, 6, 7 and 8, wherein the cell-binding agent T is selected from an antibody; a single chain antibody; an antibody fragment that binds to the target cell; a monoclonal antibody; a single chain monoclonal antibody; or a monoclonal antibody fragment that binds to the 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; adnectins that mimic antibodies; DARPins; a lymphokine; a hormone; a vitamin; a growth factor; a colony stimulating factor; a nutrient-transport molecule (a transferrin); or a binding peptide, protein, small molecule attached on albumin, a polymer, a dendrimer, a liposome, a nanoparticle, a vesicle, a (viral) capsid.

### Claim 11 (depends on 1, 2, 3, 4, 5, 6, 7, 8)

The conjugate of the cytotoxic agents according to claims 1, 2, 3, 4, 5, 6, 7 and 8 comprises, the formula of IIa, IIb, IIc, IId, IIe, IIf, IIg, IIh, IIi, IIj, IIk, IIl, IIm, IIn, IIo, IIp, IIq, IIr, IIIa, IIIb, IIIc, IIId, IIIe, IIIf, IIIg, IIIh, IIIi, IIIj, IVa, IVb, IVc, IVd, IVe, IVf, Va, Vb, Vc, Vd, Ve, Vf, Vg, Vh, VIa, VIb, VIc, VIe, VIf, VIg, VIh, VIi, VIIa, VIIb, VIIc, VIId, VIIe, VIIf, VIIg, VIIh, VIIi, VIIj, VIIk, VIIl, VIIm, VIIn, VIIo, VIIp, VIIr, VIIs, and VIIt, which are illustrated below:
Wherein Aa is a natural or unnatural amino acid; Ar is aryl; n is 1˜20; q=1˜5; X′, Y′ and Z′ are independently CH, O, S, NH, or NR22; R22, R23 and R24 are independently C1˜C8 of alkyl; C2˜C8 of alkenyl, alkynyl, heteroalkyl; C3˜C8 of aryl, heterocyclic, carbocyclic, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, Ar-alkyl, heteroalkylcycloalkyl, heteroaralkyl, or —(OCH2CH2)n—; R′ and R″ are independently H or CH3. Inside the round brackets are the antimitotic drugs and the inside square brackets are the antimitotic drug conjugated to a cell binding agent T via a linker.

### Claim 12 (depends on 1, 2, 3, 4, 5, 6, 7, 8)

The conjugate of the cytotoxic agents according to claims 1, 2, 3, 4, 5, 6, 7 and 8 comprises antimitotic agents and linkers having structures of 232a, 232b, 232c, 232d, 232e, 232f, 232g, 232h, 232i, 232j, 232k, 2321, 232m, 232n, 232o, 232p, 232q, 232r, 232s, 232t, 221a, 221b, 233a, 233b, 233c, 233d, 222a, 308a, 308b, 309a, 309b, 290, 299a, 299b, 241, 242, 247, 248, 206, 211, 346, 350, 354, 357, 361, 366, 371, and 376 which are illustrated below:

### Claim 13 (depends on 1, 2, 3, 4, 5, 6, 7, 8)

The conjugate of the cytotoxic agents according to claims 1, 2, 3, 4, 5, 6, 7 and 8 comprises Formula of: mAb-TZ01, mAb-TZ02, mAB-TZ03, mAb-TZ04, mAb-TZ05, mAb-TZ06, mAb-TZ07, mAb-TZ08, mAb-TZ09, mAb-TZ10a, mAb-TZ10b, mAb-5 TZ11, mAb-TZ12, mAb-TZ13, mAb-TZ14, mAb-TZ15, mAb-TZ16, mAb-TZ17, mAb-TZ18, mAb-TZ19, mAb-TZ20, mAb-TZ21, mAb-TZ22, mAb-TZ23, mAb-TZ24, and mAb-TZ25 which are illustrated below:
Wherein mAb is an antibody.

### Claim 14 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)

The conjugate of the cytotoxic agents according to claims 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, and 13 is used to target cells comprising tumor cells, virus infected cells, microorganism infected cells, parasite infected cells, autoimmune cells, activated cells, myeloid cells, activated T-cells, B cells, or melanocytes, cells expressing one or more of Apo2, ASLG659, BAFF-R, BMPR1B (bone morphogenetic protein receptor), IGF-IR, CA125, CanAg, E16, EGFR, EphA2 receptor, ErbB2, interleukin, MUC1, MUC16, Napi3b (NAPI-3B, NPTIIb, SLC34A2, solute carrier family 34, member 2, type II sodium-dependent phosphate transporter 3b), VEGF, TF, MY9, anti-B4, EpCAM, FcRH2, C242, CD2, CD3, CD4, CD5, CD6, CD11, CD 11a, CD18, CD19, CD20, CD21, CD22, CD26, CD30, CD33, CD37, CD38, CD40, CD44, CD56, CD70, CD72, CD79, CD79a, CD79b, CD90, CD105, CD138, CRIPTO (CR, CR1, CRGF, CRIPTO, CXCR5, LY64, TDGF1, Tetratocarcinoma-derived growth factors), endothelin B receptor (ETBR), EphA receptors, EphB receptors, EGFr, EGFRvIII, ETBR (Endothelin), FCRH1, HER2, HER2/neu, HER3, HLA-DOB (MHC class II molecule Ia antigen), integrins, IRTA2, LIV-1, MPF (MPF, MSLN, SMR, megakaryocyte potentiating factor, mesothelin), GEDA, Mesothelin, NaPi2b, PDL1, Sema 5b (FLJ10372, KIAA1445, Mm42015, SEMA5B, 5EMAG, semaphoring 5 bHlog, sdema domain, seven thrombospondin repeats, cytoplasmic domain), PSCA, PSMA, STEAP1 (six transmembrane epithelial antigen of prostate 1), and STEAP2 (HGNC 8639, IPCA-1, PCANP1, STAMP1, STEAP2, STMP, prostate) antigens; and cells expressing insulin growth factor receptor, epidermal growth factor receptor, or folate receptor.

### Claim 15 (depends on 9)

The conjugate according to claim 9, wherein when the linker is used to conjugate two cytotoxic agents, having a 3-bromo-maleimido, or 3, 4-dibromo-maleimido terminus as illustrated in the following formula:
Wherein R is C1˜C8 alkyl or C2H4(OC2H4)n, n is 1˜20; X3 is H or Br.

### Claim 16 (depends on 1, 2, 3, 4, 5, 6, 7, 8)

The conjugate of the cytotoxic agents according to claims 1, 2, 3, 4, 5, 6, 7, 8, and 9, wherein when two cytotoxic agents are linked to a linker, having the following formula:
Wherein R is C1˜C8 alkyl or C2H4(OC2H4)n, n is 1˜20; D is a cytotoxic agent/drug, T is a cell binding molecule.

### Claim 17 (depends on 1, 2, 3, 4, 5, 6, 7, 8)

The conjugate of the cytotoxic agents according to claims 1, 2, 3, 4, 5, 6, 7, 8, and 9, wherein when two cytotoxic agents are linked to a linker, having the following formula:
Wherein R is C1˜C8 alkyl or C2H4(OC2H4)n, n is 1˜20; D is a cytotoxic agent/drug, T is a cell binding molecule.

### Claim 18 (depends on 16, 17)

The conjugate of claims 16 and 17, wherein the cytotoxic drugs D can also be selected from chemical compounds useful in the treatment of cancer, such as calicheamicins, auristatins, maytansinoids, dolastatins, CC-1065 analogs, doxorubicins, taxanes, pyrrolobenzodiazepine dimmer, siRNA or a combination of them thereof, and the conjugate can also be pharmaceutically acceptable salts, acids or derivatives of any of the above.

### Claim 19 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 11)

A pharmaceutical composition comprising a therapeutically effective amount of the conjugate of a cell binding molecule—cytotoxic agents according to claims 1, 2, 3, 4, 5, 6, 7, 8, 11, or 13, or its pharmaceutically acceptable salt, and carrier, diluent, or excipient thereof, or a combination of them thereof, for the treatment or prevention of cancer, autoimmune disease, or infectious disease.

### Claim 20 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 11)

A pharmaceutical composition comprising a therapeutically effective amount of the conjugate of a cell binding molecule cytotoxic agents according to claims 1, 2, 3, 4, 5, 6, 7, 8, 11, or 13, administered concurrently with the other therapeutic agents such as chemotherapeutic agent, radiation therapy, immunotherapy agents, anti-autoimmune disorder agents, anti-infectious agents or other antibody-drug conjugates for synergistical treatment or prevention of a cancer, an autoimmune disease, or an infectious disease.