Specific conjugation linkers, specific immunoconjugates thereof, methods of making and uses such conjugates thereof

Patent metadata

Field	Value
Publication number	US-12478695-B2
Application number	US-202117373950-A
Country	US
Kind code	B2
Filing date	Jul 13, 2021
Priority date	Feb 4, 2016
Publication date	Nov 25, 2025
Grant date	Nov 25, 2025

Abstract

The present invention relates to novel linkers containing a 2,3-disubstituted succinic group, or 2-monosubstituted, or 2,3-disubstituted fumaric or maleic (trans (E)- or cis (Z)-butenedioic), or acetylenedicarboxyl group for conjugation of a cytotoxic agent, and/or one or more different functional molecules per linker to a cell-binding molecule, through bridge linking pairs of thiols on the cell-binding molecule specifically. The invention also relates to methods of making such linkers, and of using such linkers in making homogeneous conjugates, as well as of application of the conjugates in treatment of cancers, infections and autoimmune disorders.

First claim

What is claimed is:

1 . A compound having following formula (I):

wherein:
represents either a single bond or a double bond;
R 1 is absent, or selected from the group consisting of C 1 -C 8 of alkyl; C 2 -C 8 of heteroalkyl, alkylcycloalkyl, or heterocycloalkyl; C 3 -C 8 of aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl; C 1 -C 8 of ester, ether, or amide; polyethyleneoxy unit of formula (OCH 2 CH 2 ) p or (OCH 2 CH(CH 3 )) p , wherein p is an integer from 0 to about 1000, and a combination thereof; or R 1 is a chain of atoms selected from the group consisting of C, N, O, S, Si, and P;
X 1 is selected from the group consisting of O; S; B(R 3 ); Si(R 3 )N(R 3′ ); P(O)(R 3 ); C 2 -C 8 of heterocycloalkyl; C 3 -C 8 of aryl, heterocyclic, or heteroaryl; 1-8 amino acids; wherein R 3 and R 3′ are independently H; C 1 -C 8 of alkyl; C 2 -C 8 of heteroalkyl, alkylcycloalkyl, or heterocycloalkyl; C 3 -C 8 of aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl; C 1 -C 8 of ester, ether, or amide; polyethyleneoxy unit of formula (OCH 2 CH 2 ) p or (OCH 2 CH(CH 3 )) p , wherein p is an integer from 0 to about 1000, and a combination thereof,
X 2 is absent or selected from the group consisting of NH; NHNH; N(R 3 ); N(R 3 )N(R 3′ ); O; S; B(R 3 ); Si(R 3 )N(R 3 ); P(O)(R 3 ); C 1 -C 8 of alkyl; C 2 -C 8 of heteroalkyl, alkylcycloalkyl, or heterocycloalkyl; C 3 -C 8 of aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl; 1-8 amino acids; wherein R 3 and R 3′ are independently H; C 1 -C 8 of alkyl; C 2 -C 8 of heteroalkyl, alkylcycloalkyl, or heterocycloalkyl; C 3 -C 8 of aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl; C 1 -C 8 of ester, ether, or amide; polyethyleneoxy unit of formula (OCH 2 CH 2 ) p or (OCH 2 CH(CH 3 )) p , wherein p is an integer from 0 to about 1000, and a combination thereof,
U and U′ are independently H or a leaving group that can be substituted by a thiol, provided that when represents a single bond, both U and U′ are not H; and when represents a double bond, at least one of U and U′ is the leaving group;
R 2 is independently selected from the group consisting of OH, H, NH 2 ; SH; NHNH 2 ; N(R 3 )(R 3′ ); N(R 3 )NH(R 3′ ); polyethyleneoxy unit of 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 , or CH 2 CH 2 (OCH 2 CH 2 ) p COOR 3 ,wherein p and p′ are independently an integer selected from 0 to about 1000, or a combination thereof, C 1 -C 8 of alkyl; C 2 -C 8 of heteroalkyl, alkylcycloalkyl, or heterocycloalkyl; and C 3 -C 8 of aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl; wherein R 3 and R 3′ are independently H; C 1 -C 8 of alkyl; C 2 -C 8 of heteroalkyl, alkylcycloalkyl, or heterocycloalkyl; C 3 -C 8 of aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl; C 1 -C 8 of ester, ether, or amide; 1-8 amino acids; polyethyleneoxy unit of formula (OCH 2 CH 2 ) p or (OCH 2 CH(CH 3 )) p , wherein p is an integer from 0 to about 1000, or a combination thereof; or a chain of atoms selected from the group consisting of C, N, O, S, Si, and P;
m 1 and m 2 are independently an integer from 1 to 30; and
Z 1 is a function group that enables the compound to react with a cytotoxic drug, to form a disulfide, thioether, thioester, peptide, hydrazone, ether, ester, carbamate, carbonate, secondary, tertiary, or quaternary amine, imine, cycloheteroalkyane, heteroaromatic, alkoxime or amide bond, and comprises at least one of thiol, disulfide, carboxyl, aldehyde, ketone, N-hydroxysuccinimide ester, p-nitrophenyl ester, dinitrophenyl ester, pentafluorophenyl ester, carboxylic acid chloride, carboxylic acid anhydride, pyridyldisulfide, nitropyridyldisulfide, maleimide, haloacetate, methylsulfonephenyloxadiazole, amine, alkoxyamine, hydrazine, acyloxylamine, hydrazide, or alkyne.

2 . The compound of claim 1 , wherein the leaving group is a halide, methanesulfonyl, toluenesulfonyl, trifluoromethyl-sulfonyl, trifluoromethylsulfonate, nitrophenoxyl, N-succinimidyloxyl, phenoxyl; dinitrophenoxyl; pentafluorophenoxyl, tetrafluorophenoxyl, trifluorophenoxyl, difluorophenoxyl, monofluorophenoxyl, pentachlorophenoxyl, 1H-imidazole-1-yl, chlorophenoxyl, dichlorophenoxyl, trichlorophenoxyl, tetrachlorophenoxyl, N-(benzotriazolyl)oxyl, 2-ethyl-5-phenylisoxazolium-yl, 2-ethyl-5-phenylisoxazolium-3′-sulfonyl, phenyloxadiazolyl, phenyloxadiazole-sulfonyl, or oxadiazolyl.

3 . The compound of claim 1 , wherein the leaving group is a halide, methanesulfonyl, p-toluenesulfonyl, trifluoromethyl sulfonyl, and trifluoromethylsulfonate.

4 . The compound of claim 1 , wherein the leaving group is bromide or iodide.

5 . The compound of claim 1 , wherein: (i) represents a single bond, both U and U′ are bromide or iodide; (ii) represents a double bond, both U and U′ are bromide or iodide; or (iii) represents a double bond, one of U and U′ is bromide or iodide and the other is H.

6 . The compound of claim 1 , wherein the component

is 2,3-disubstituted succinic group, 2-monosubstituted or 2,3-disubstitutedfumaric group, or 2-monosubstituted or 2,3-disubstitutedmaleic group, and is capable of reacting with a pair of thiols of a cell-binding agent.

7 . The compound of claim 1 , wherein m 1 and m 2 are independently an integer from 1 to 10.

8 . The compound of claim 1 , wherein R 1 , R 3 or R 3′ comprises alkane, alkenylene, alkynylene, ether, polyoxyalkylene, ester, amine, imine, polyamine, hydrazine, hydrazone, amide, urea, semicarbazide, carbazide, alkoxyamine, alkoxylamine, urethane, amino acid, peptide, acyloxylamine, hydroxamic acid, or a combination of two or more thereof.

9 . The compound of claim 1 , wherein R 1 , R 2 , R 3 or R 3′ is a linear alkyl having from 1-6 carbon atoms, or polyethyleneoxy unit of formula (OCH 2 CH 2 ) p , p=1-100.

10 . The compound of claim 1 , wherein R 1 or R 2 has independently one or more, or repeating, or a combining thereof, of components depicted in the following:

and L- or D-, natural or unnatural peptides containing 1-20 amino acids.

11 . The compound of claim 1 , wherein R 1 or R 2 is cleavable by a protease.

12 . The compound of claim 1 , wherein Z 1 comprises at least one of N-hydroxysuccinimide ester, p-nitrophenyl ester, dinitrophenyl ester, pentafluorophenyl ester, carboxylic acid chloride, carboxylic acid anhydride, pyridyldisulfide, nitropyridyldisulfide, maleimide, haloacetate, methylsulfonephenyloxadiazole, amine, alkoxyamine, hydrazine, acyloxylamine, hydrazide,

Assignees

• Hangzhou Dac Biotech Co Ltd
• Zhao Robert Yongxin

Inventors

• Zhao Robert Yongxin
• Yang Qingliang
• Huang Yuanyuan
• Gai Shun
• Zhao Linyao
• Ye Hangbo
• Guo Huihui
• Tong Qianqian
• Cao Minjun
• Jia Junxiang
• Yang Chengyu
• Li Wenjun
• Zhou Xiaomai
• Xie Hongsheng
• Lin Chen
• Guo Zhixiang
• Ye Zhicang

Classifications

• A61K47/68035

  the drug being a pyrrolobenzodiazepine · CPC title

• A61K47/68033

  the drug being a maytansine · CPC title

• A61K47/68031

  the drug being an auristatin · CPC title

• A61K31/194

  having two or more carboxyl groups, e.g. succinic, maleic or phthalic acid · CPC title

• C07D519/00

  Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00 · CPC title