Search

CN-122003251-A - TROP 2-targeted antibody-drug conjugate and preparation method and application thereof

CN122003251ACN 122003251 ACN122003251 ACN 122003251ACN-122003251-A

Abstract

Provided are an antibody-drug conjugate targeting TROP2, and a preparation method and application thereof. The TROP 2-targeted antibody-drug conjugate shows strong tumor-specific cytotoxicity and shows stronger 'bystander effect' in vivo experiments, and therefore, the TROP 2-targeted antibody-drug conjugate can be used as a medical drug, particularly used as a single drug or combined drug for treating tumors, used for new adjuvant treatment of tumors and the like.

Inventors

  • XIONG HUIZHONG
  • ZHANG XIGUANG
  • ZHU CHENCHEN

Assignees

  • 信达生物制药(苏州)有限公司

Dates

Publication Date
20260508
Application Date
20240927
Priority Date
20230928

Claims (20)

  1. An antibody-drug conjugate represented by the following formula (I), a stereoisomer or a pharmaceutically acceptable salt or solvate thereof, Wherein D is represented by the following formula (II): Wherein R 1 is selected from H, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 1 -C 6 haloalkyl, C 2 -C 6 haloalkenyl, and C 2 -C 6 haloalkynyl; R 2 is selected from H, halogen, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, -OR 6 , and-SR 6 ;R 3 is selected from H, halogen, CN, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, and-OR 6 , OR R 2 and R 3 together form-O (CH 2 ) n O-OR-O (CF 2 ) n O-, wherein n is 1 OR 2; R 6 is selected from H or C 1 -C 4 alkyl, and L 1 is-L 1b -L 1a -, wherein L 1b is linked to Q', L 1a is linked to D, Wherein L 1a is absent or- (C 1 -C 10 alkylene) -; l 1b is absent, - (C 1 -C 10 alkylene) -C (O) N (R 5 ) -or- (C 1 -C 10 alkylene) -N (R 5 ) C (O) -, wherein R 5 is H or C 1 -C 6 alkyl, Q' is-O-or-S-; E is-CH 2 -NH-M, said M being a peptide residue comprising 2 to 10 amino acids, wherein said peptide residue is optionally substituted with 1 or more groups independently selected from C 1-6 alkyl and a polyol group, and wherein the N-terminus of M is covalently attached to Z'; Z 'is-C (=O) -L 2 -Y' -, wherein Z 'is covalently linked to E through the-C (=O) moiety shown and to A through the Y' moiety; L 2 is selected from -(CH 2 ) m -(O-CH 2 CH 2 ) m1 -(CH 2 ) m2 -、-(CH 2 ) m -(O-CH 2 CH 2 ) m1 -NHC(O)-(CH 2 ) m -、-(CH 2 ) m -(O-CH 2 CH 2 ) m1 -C(O)NH-(CH 2 ) m -, wherein m1 and m2 are independently selected from integers from 0 to 20, m is selected from integers from 1 to 10, wherein the left end of these groups is attached to-C (=O) -and the right end is attached to Y'; Y' is composed of A group formed; A represents an antibody or antibody fragment targeting TROP2 comprising a heavy chain comprising a CDR1 comprising or consisting of the sequence shown in amino acid sequence SEQ ID NO. 1, a CDR2 comprising or consisting of the sequence shown in amino acid sequence SEQ ID NO. 2, and a CDR3 comprising or consisting of the sequence shown in amino acid sequence SEQ ID NO. 3, The light chain comprises a CDR1 comprising or consisting of the sequence shown in amino acid sequence SEQ ID NO. 4, a CDR2 comprising or consisting of the sequence shown in amino acid sequence SEQ ID NO. 5, and a CDR3 comprising or consisting of the sequence shown in amino acid sequence SEQ ID NO. 6, and P is the average of the drug to antibody ratio and is a value between 1 and 15, for example between 3 and 5, 3.5 and 4.5.
  2. The antibody-drug conjugate, stereoisomer or pharmaceutically acceptable salt or solvate thereof according to claim 1, wherein the TROP 2-targeting antibody or antibody fragment comprises a variable domain of a heavy chain and a variable domain of a light chain, wherein the variable domain of the heavy chain has at least 80% sequence identity, preferably at least 90% sequence identity, such as at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or even 100% sequence identity with the amino acid sequence SEQ ID No. 7 and the variable domain of the light chain has at least 80%, preferably at least 90% sequence identity, such as at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or even 100% sequence identity with the amino acid sequence SEQ ID No. 8.
  3. The antibody-drug conjugate, stereoisomer or pharmaceutically acceptable salt or solvate thereof of claim 1 or 2, wherein the TROP 2-targeting antibody or antibody fragment comprises a variable domain of a heavy chain and a variable domain of a light chain, wherein the variable domain of the light chain comprises or consists of the amino acid sequence shown in SEQ ID No. 8 and the variable domain of the heavy chain comprises or consists of the amino acid sequence shown in SEQ ID No. 7.
  4. The antibody-drug conjugate, stereoisomer or pharmaceutically acceptable salt or solvate thereof of any of claims 1-3, wherein the TROP 2-targeting antibody or antibody fragment comprises a heavy chain and a light chain, wherein the light chain of the TROP 2-targeting antibody or fragment thereof has at least 80% sequence identity, preferably at least 90% sequence identity, such as at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or even 100% sequence identity with the amino acid sequence SEQ ID No. 10, and the heavy chain of the TROP 2-targeting antibody or fragment thereof has at least 80% sequence identity, preferably at least 90% sequence identity, such as at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or even 100% sequence identity with the amino acid sequence SEQ ID No. 9.
  5. The antibody-drug conjugate, stereoisomer, or pharmaceutically acceptable salt or solvate thereof of any one of claims 1-4, wherein the TROP 2-targeting antibody or antibody fragment comprises a heavy chain and a light chain, wherein the heavy chain comprises or consists of the amino acid sequence set forth in SEQ ID No. 9, and the light chain comprises or consists of the amino acid sequence set forth in SEQ ID No. 10.
  6. The antibody-drug conjugate, stereoisomer or pharmaceutically acceptable salt or solvate thereof of any one of claims 1-5, wherein the antibody fragment is selected from Fab, fab ', F (ab ') 2, fab ' -SH, scFv or scFv-Fc.
  7. The antibody-drug conjugate of any one of claims 1-6, a stereoisomer, or a pharmaceutically acceptable salt or solvate thereof, wherein the TROP 2-targeting antibody is a monoclonal antibody.
  8. The antibody-drug conjugate, stereoisomer, or pharmaceutically acceptable salt or solvate thereof of any one of claims 1-6, wherein the TROP 2-targeting antibody is a chimeric or humanized antibody.
  9. The antibody-drug conjugate of any of claims 1-8, a stereoisomer or a pharmaceutically acceptable salt or solvate thereof, Wherein R 1 is selected from-H or C 1 -C 4 alkyl, R 2 is selected from-H, -F, C 1 -C 3 alkyl or C 1 -C 3 haloalkyl, R 3 is selected from-H, -F, -CN, -OCH 3 、-CH 3 or-CF 3 ; L 1 is independently unsubstituted or halogenated- (C 1 -C 10 alkylene) -; q' is-O-or-S-; E is-CH 2 -NH-M, said M being a peptide comprising 2 to 10 amino acids, wherein optional amino acids are substituted with one or more polyols, and wherein the N-terminus of said M is covalently attached to Z'; Z 'is-C (=O) -L 2 -Y'; L 2 is- (C 1 -C 10 alkylene) -.
  10. The antibody-drug conjugate, stereoisomer or pharmaceutically acceptable salt or solvate thereof according to any one of claims 1-9, wherein Y' is Wherein represents the site of covalent attachment to the a.
  11. The antibody-drug conjugate of any of claims 1-10, a stereoisomer or a pharmaceutically acceptable salt or solvate thereof, Wherein L 2 is- (CH 2 ) m ) -and m is an integer selected from 1 to 10.
  12. The antibody-drug conjugate, stereoisomer or pharmaceutically acceptable salt or solvate thereof according to any one of claims 1-11, wherein Z' is formed from:
  13. The antibody-drug conjugate, stereoisomer or pharmaceutically acceptable salt or solvate thereof according to any one of claims 1-12, wherein Z' is: Wherein represents the site of covalent attachment to a.
  14. The antibody-drug conjugate, stereoisomer or pharmaceutically acceptable salt or solvate thereof according to any one of claims 1-13, wherein M is a peptide comprising 2,3 or 4 amino acids; Each amino acid of M is an L amino acid; or at least one amino acid of said M is a D amino acid.
  15. The antibody-drug conjugate, stereoisomer, or pharmaceutically acceptable salt or solvate thereof according to any one of claims 1-13, wherein the amino acid is selected from glycine, alanine, valine, glutamine, glutamic acid, phenylalanine, leucine, tyrosine, lysine, citrulline, serine, tryptophan, aspartic acid, asparagine, isoleucine, arginine, and proline, and wherein the glutamine or glutamic acid is optionally substituted with 1 polyol group and optionally substituted with 1C 1-6 alkyl group.
  16. The antibody-drug conjugate, stereoisomer, or pharmaceutically acceptable salt or solvate thereof according to any one of claims 1-15, wherein the M comprises one or more amino acids selected from glycine, alanine, valine, glutamine, glutamic acid, phenylalanine, and leucine, and wherein the glutamine or glutamic acid is optionally substituted with a polyol.
  17. The antibody-drug conjugate of any of claims 1-15, a stereoisomer or a pharmaceutically acceptable salt or solvate thereof, The substituted glutamine or glutamic acid has the structure shown below: Wherein R 4 is H or C 1 -C 6 alkyl; Preferably is Wherein R 4 is H or C 1 -C 6 alkyl.
  18. The antibody-drug conjugate of claim 17, a stereoisomer or a pharmaceutically acceptable salt or solvate thereof, The substituted glutamine or glutamic acid has the structure shown below: Wherein R 4 is H or C 1 -C 6 alkyl; Preferably is Wherein R 4 is H or C 1 -C 6 alkyl.
  19. The antibody-drug conjugate, stereoisomer or pharmaceutically acceptable salt or solvate thereof according to any one of claims 1-17, wherein M comprises an amino acid having the structure, Wherein R 4 is-H or C 1 -C 6 alkyl.
  20. The antibody-drug conjugate, stereoisomer or pharmaceutically acceptable salt or solvate thereof according to claim 19, wherein M comprises an amino acid having the structure

Description

TROP 2-targeted antibody-drug conjugate and preparation method and application thereof Technical Field The application relates to an antibody-drug conjugate (ADC), in particular to an antibody-drug conjugate targeting TROP2, a preparation method and application thereof. Background ADCs (antibody drug conjugates) are typically composed of three parts, an antibody, a payload (e.g., a biologically active small molecule), and a Linker (Linker). The load is coupled to the antibody in a covalent bond form through a linker, the antibody (such as a monoclonal antibody) can specifically identify a specific target point on the surface of a tumor cell, and then the antibody can guide the ADC to reach the surface of the cancer cell and enable the ADC to enter the cancer cell through an endocytosis effect, and then the load is released in a tumor microenvironment, so that the cancer cell is specifically killed without damaging normal tissues. TROP2, a trophoblastic cell surface antigen, also known as tumor-associated calcium signaling protein (TACSTD 2) is overexpressed in a variety of human epithelial cancers, including breast, lung, stomach, colorectal, pancreatic, prostate, cervical, head and neck, and ovarian cancers, and the like (Yezhe Cheng et al, frontiers in Oncology,2022 Dec 23;12:951589). Preclinical and clinical studies have demonstrated the use of anti-TROP 2 antibody-drug conjugates (ADCs) (e.g., trodelvy, anti-human TROP2 antibody-SN-38 conjugates) for the treatment of cancer. Clinical results show that the Trodelvy can achieve good therapeutic effects in treating refractory solid tumors. The Objective Remission Rate (ORR) of Trodelvy reached 33% in drug resistant Triple Negative Breast Cancer (TNBC) patients. Trodelvy one of the important mechanisms of action, SN-38 (payload) is linked by a pH sensitive linker that can break and specifically release SN-38 in acidic tumor microenvironments. However, because the Trodelvy linker is not sufficiently stable, maleimide-mediated linkers break under physiological conditions through thiol exchange, resulting in a relatively short half-life of Trodelvy in serum (about 1 day). Therefore Trodelvy may have a relatively high off-target effect. There remains a need in the art for TROP 2-targeting antibody-drug conjugates for use in therapy. Disclosure of Invention Accordingly, one aspect of the present application is to provide an antibody-drug conjugate having the following formula (I), a stereoisomer or a pharmaceutically acceptable salt or solvate thereof Wherein D is represented by the following formula (II): Wherein R 1 is selected from H, C 1-C6 alkyl, C 2-C6 alkenyl, C 2-C6 alkynyl, C 1-C6 haloalkyl, C 2-C6 haloalkenyl, and C 2-C6 haloalkynyl; R 2 is selected from H, halogen, C 1-C6 alkyl, C 1-C6 haloalkyl, -OR 6, and-SR 6;R3 is selected from H, halogen, CN, C 1-C6 alkyl, C 1-C6 haloalkyl, and-OR 6, OR R 2 and R 3 together form-O (CH 2)n O-OR-O (CF 2)n O-, wherein n is 1 OR 2; R 6 is selected from H or C 1-C4 alkyl, and L 1 is-L 1b-L1a -, wherein L 1b is linked to Q', L 1a is linked to D, Wherein L 1a is absent or- (C 1-C10 alkylene) -; l 1b is absent, - (C 1-C10 alkylene) -C (O) N (R 5) -or- (C 1-C10 alkylene) -N (R 5) C (O) -, wherein R 5 is H or C 1-C6 alkyl, Q' is-O-or-S-; E is-CH 2 -NH-M-said M being a peptide residue comprising 2 to 10 amino acids, wherein said peptide residue is optionally substituted with 1 or more (e.g., 2,3 or 4) groups independently selected from C 1-6 alkyl and a polyol group, and wherein the N-terminus of M is covalently attached to Z'; Z 'is-C (=O) -L 2 -Y' -, wherein Z 'is covalently linked to E through the-C (=O) moiety shown and to A through the Y' moiety; L 2 is selected from -(CH2)m-(O-CH2CH2)m1-(CH2)m2-、-(CH2)m-(O-CH2CH2)m1-NHC(O)-(CH2)m-、-(CH2)m-(O-CH2CH2)m1-C(O)NH-(CH2)m-, wherein m1 and m2 are independently selected from integers from 0 to 20, for example 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16, m is selected from integers from 1 to 10, for example 1, 2, 3, 4, 5, 6, 7 or 8, wherein the left end of these groups is attached to-C (=O) -and the right end is attached to Y'; Y' is composed of A group formed; a represents an antibody or antibody fragment, preferably an antigen binding fragment, targeting TROP2, and P is the average of the drug to antibody ratios (i.e., average DAR) and is a value between 1 and 15, for example, between 2 and 10, 2 and 8, 2 and 6, 2 and 5, 3 and 5, and 3.5 and 4.5. In one embodiment, the application provides a TROP 2-targeting antibody-drug conjugate having the structure of formula (I): wherein D is represented by the following formula (II): Wherein R 1 is selected from-H or C 1-C4 alkyl, R 2 is selected from-H, -F, C 1-C3 alkyl or C 1-C3 haloalkyl, R 3 is selected from-H, -F, -CN, -OCH 3、-CH3 or-CF 3; L 1 is independently unsubstituted or halogenated- (C 1-C10 alkylene) -; q' is-O-or-S-; E is-CH 2 -NH-M, said M being a peptide comprising 2 to 10 amino acids, wherein optional ami