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

Patent metadata

Field	Value
Publication number	US-11779663-B2
Application number	US-202017093184-A
Country	US
Kind code	B2
Filing date	Nov 9, 2020
Priority date	Feb 4, 2016
Publication date	Oct 10, 2023
Grant date	Oct 10, 2023

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

The invention claimed is:

1. A bridge linker compound of Formula (I)

wherein:
“ ” represents a triple bond;
provided that the component:

which is acetylenedicarboxyl group;
R 1 is selected from the group consisting of C 1 -C 8 alkyl; C 2 -C 8 heteroalkyl, alkylcycloalkyl, or heterocycloalkyl; C 3 -C 8 aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl; C 1 -C 8 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, which covalently connects to X 1 and Z 1 ;
X 1 and X 2 are independently 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 alkyl; C 2 -C 8 heteroalkyl, alkylcycloalkyl, or heterocycloalkyl; C 3 -C 8 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 alkyl; C 2 -C 8 heteroalkyl, alkylcycloalkyl, or heterocycloalkyl; C 3 -C 8 aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl; C 1 -C 8 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,
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 alkyl; C 2 -C 8 heteroalkyl, alkylcycloalkyl, or heterocycloalkyl; and C 3 -C 8 aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl; wherein R 3 and R 3 ′ are independently H; C 1 -C 8 alkyl; C 2 -C 8 heteroalkyl, alkylcycloalkyl, or heterocycloalkyl; C 3 -C 8 aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl; C 1 -C 8 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,
m 1 and m 2 are independently an integer from 1 to 30;
Z 1 is at least one of thiol, disulfide, amino, carboxyl, aldehyde, ketone, maleimido, haloacetyl, hydrazines, alkoxyamino, hydroxy, N-hydroxysuccinimide ester, p-nitrophenyl ester, dinitrophenyl ester, pentafluorophenyl ester, carboxylic acid chloride, carboxylic acid anhydride, pyridyldisulfide, nitropyridyldisulfide, maleimide, haloacetate, methylsulfonephenyloxadiazole, acyloxylamine, hydrazide, or alkyne.

2. The bridge linker compound according to claim 1 , wherein R 1 is at least one of alkylene, alkenylene, alkynylene, ether, polyoxyalkylene, ester, amine, imine, polyamine, hydrazine, hydrazone, amide, urea, semicarbazide, carbazide, alkoxyamine, alkoxylamine, urethanes, amino acid, peptide, acyloxylamine, or hydroxamic acid.

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

4. The bridge linker compound according to claim 1 , wherein R 1 or R 2 is cleavable by a protease.

5. The bridge linker compound according to 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.

6. The bridge linker compound according to claim 1 , wherein m 1 and m 2 are independently an integer from 1 to 10.

7. The bridge linker compound of claim 1 , wherein Z 1 is

wherein X 1 ′ is F, Cl, Br, I or Lv 3 ; X 2 ′ is O, NH, N(R 1 ), or CH 2 ; R 3 and R 5 are H, R 1 , aromatic, heteroaromatic, or aromatic group wherein one or several H atoms are replaced independently by —R 1 , -halogen, —OR 1 , —SR 1 , —NR 1 R 2 , —NO 2 , —S(O)R 1 , —S(O) 2 R 1 , or —COOR 1 ; Lv 3 is a leaving group selected from the group consisting of 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, phenyloxadiazol-yl, oxadiazol-yl, and an intermediate molecule generated with a condensation reagent for Mitsunobu reactions, wherein R 1 and R 2 are the same defined in claim 1 .

8. A method for preparing the bridge linker compound of Formula (I) of claim 1 , comprising a step of forming the acetylenedicarboxyl group, by condensation of an acetylenedicarboxyl compound, or an acid derivative thereof, with components containing a 1° or 2° amine, alcohol, or thiol, on their terminal as shown in the following scheme (Ia) or (Ib):

wherein X 1 , X 2 , R 1 , and R 2 are described the same in claim 1 ;
W and W′ are the same or independently a 1° or 2° amine, hydroxyl, or thiol;
Lv 1 and Lv 2 are the same or independently H, OH, F, Cl, Br, I, NH 2 , NHNH 2 , methane-sulfonyl, toluenesulfonyl, trifluoromethylsulfonyl, trifluoromethyl-sulfonate, nitrophenoxyl, N-succinimidyloxyl, phenoxyl; dinitrophenoxyl; pentafluo-rophenoxyl, tetrafluorophenoxyl, trifluorophenoxyl, difluorophenoxyl, monofluorophenoxyl, pentachlorophenoxyl, 1H-imidazole-1-yl, chlorophenoxyl, dichlorophenoxyl, trichlorophenoxyl, tetrachlorophenoxyl, N-(benzotriazol-yl)oxyl, 2-ethyl-5-phenylisoxazolium-3′-sulfonyl, phenyl-oxadiazole-sulfonyl, oxadiazol-yl, or an intermediate molecule generated with a condensation reagent for Mitsunobu reactions wherein the condensation reagents are: N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide, dicyclohexyl-carbodiimide), N,N′-diisopropylcarbodiimide, N-cyclohexyl-N′-(2-morpholinoethyl)carbodiimide metho-p-toluenesulfonate, 1,1′-carbonyldiimidazole, O-(benzotria-zol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate, N,N,N′,N′-tetramethyl-O-(1H-benzotriazol-1-yl)uronium hexafluorophosphate, (benzotriazol-1-yloxy)tris(dimethyl-amino)phosphonium hexafluorophosphate, (benzotriazol-1-yloxy)tripyrrolidinophos-phonium hexafluorophosphate, diethyl cyanophosphonate, chloro-N,N,N′,N′-tetrameth

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

• A61K49/0058 Primary

  Antibodies · CPC title

• A61K47/6809

  Antibiotics, e.g. antitumor antibiotics anthracyclins, adriamycin, doxorubicin or daunomycin · CPC title