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CN-122005843-A - Bispecific antibody drug conjugate for resisting B7H3 and PD-L1, and preparation method and application thereof

CN122005843ACN 122005843 ACN122005843 ACN 122005843ACN-122005843-A

Abstract

The invention discloses a bispecific antibody drug conjugate, a preparation method and application thereof, wherein the structure of the bispecific antibody drug conjugate comprises a bispecific antibody resisting B7H3 and PD-L1 or an antigen binding fragment thereof, a linker unit L and a cytotoxic drug. The bispecific antibody drug conjugate has good endocytic effect, proliferation inhibition activity and tumor growth inhibition activity.

Inventors

  • LI CHENGGANG
  • YANG JUNJIE
  • ZHANG YU
  • HUA HAIQING
  • ZHU ZHONGYUAN

Assignees

  • 映恩生物科技(上海)有限公司

Dates

Publication Date
20260512
Application Date
20231227
Priority Date
20221227

Claims (20)

  1. 1. A bispecific antibody drug conjugate comprising the structure of a bispecific antibody or antigen-binding fragment thereof against B7H3 and PD-L1, a linker unit L, and a cytotoxic drug, wherein the bispecific antibody or antigen-binding fragment thereof comprises: a monoclonal antibody unit directed against PD-L1 and comprising 2 heavy chains and 2 light chains, Nanobody units which are directed against B7H3 and comprise 2 identical nanobodies, Wherein the N-terminal of the 2 nanobodies are respectively connected with the C-terminal of the Fc fragments of the 2 heavy chains of the monoclonal antibody unit through connecting peptides.
  2. 2. The bispecific antibody drug conjugate of claim 1 wherein, The light chain variable region of the monoclonal antibody unit comprises a CDR1 having an amino acid sequence of SEQ ID NO. 1, a CDR2 having an amino acid sequence of SEQ ID NO. 2, and a CDR3 having an amino acid sequence of SEQ ID NO. 3, the heavy chain variable region of the monoclonal antibody unit comprises a CDR1 having an amino acid sequence of SEQ ID NO. 5, a CDR2 having an amino acid sequence of SEQ ID NO. 6, and a CDR3 having an amino acid sequence of SEQ ID NO. 7, and the nanobody comprises a CDR1 having an amino acid sequence of SEQ ID NO. 12, a CDR2 having an amino acid sequence of SEQ ID NO. 13, and a CDR3 having an amino acid sequence of SEQ ID NO. 14.
  3. 3. The bispecific antibody drug conjugate of claim 1 or 2, wherein, The light chain variable region of the monoclonal antibody unit comprises an amino acid sequence as set forth in SEQ ID NO. 4, the heavy chain variable region of the monoclonal antibody unit comprises an amino acid sequence as set forth in SEQ ID NO. 8, and the nanobody comprises an amino acid sequence as set forth in SEQ ID NO. 15.
  4. 4. The bispecific antibody drug conjugate of any one of claim 1 to 3, wherein, The monoclonal antibodies comprise immunoglobulin constant regions, which are human IgG constant regions, e.g., human IgG1 constant regions.
  5. 5. The bispecific antibody drug conjugate of any one of claim 1-4, wherein, The light chain of the monoclonal antibody unit comprises an amino acid sequence as set forth in SEQ ID NO. 9, the heavy chain of the monoclonal antibody unit comprises an amino acid sequence as set forth in SEQ ID NO. 10, and the nanobody comprises an amino acid sequence as set forth in SEQ ID NO. 15, or The full-length amino acid sequence of the light chain of the monoclonal antibody unit is shown as SEQ ID NO. 9, the full-length amino acid sequence of the heavy chain of the monoclonal antibody unit is shown as SEQ ID NO. 10, and the amino acid sequence of the nano antibody is shown as SEQ ID NO. 15.
  6. 6. The bispecific antibody drug conjugate of any one of claims 1-5, wherein the linker peptide is a polypeptide comprising glycine and serine and having a certain elasticity and protease resistance, preferably the amino acid sequence of the linker peptide is SEQ ID No. 11.
  7. 7. The bispecific antibody drug conjugate of any one of claims 1-6, wherein the heavy chain amino acid sequence of the bispecific antibody is shown in SEQ ID No. 16 and the light chain amino acid sequence is shown in SEQ ID No. 9.
  8. 8. The bispecific antibody drug conjugate of any of claim 1-7, wherein the cytotoxic drug is a structure of formula (A-1), a tautomer, enantiomer, diastereomer, or mixture of isomers thereof, or a pharmaceutically acceptable salt or solvate thereof, ; Wherein, the M is-L 2 -L 1 -C (O) -; L 2 is-O-or-S-, preferably-O-, and L 2 is connected to said linker unit L; L 1 is- (C (R 1a )(R 1b )) m -CH 2 -、C 3 -C 6 saturated cycloalkyl or 3-6 membered saturated heterocyclyl, each of which is independently optionally substituted with one or more R 2a ; m is 1,2,3 or 4, and hetero atoms in the 3-6 membered saturated heterocyclic group are N, O or S independently, and the number of the hetero atoms is 1,2 or 3; r 1a and R 1b are each independently hydrogen, halogen, hydroxy, amino, or C 1 -C 6 alkyl, said C 1 -C 6 alkyl optionally substituted with one or more halogens; R 2a is halogen, hydroxy, amino, or C 1 -C 6 alkyl, said C 1 -C 6 alkyl optionally substituted with one or more halogens.
  9. 9. The bispecific antibody drug conjugate of claim 8, wherein, L 1 is- (C (R 1a )(R 1b )) m -CH 2 -; each R 1a is independently hydrogen, halogen or C 1 -C 6 alkyl; each R 1b is independently hydrogen, halogen or C 1 -C 6 alkyl); Preferably, L 1 is- (C (R 1a )(R 1b )) m -CH 2 -; each R 1a is independently of the others C 1 -C 6 alkyl, preferably C 1 -C 3 alkyl; each R 1b is independently of the others hydrogen or C 1 -C 6 alkyl, preferably hydrogen or C 1 -C 3 alkyl); Preferably, L 1 is- (C (R 1a )(R 1b )) m -CH 2 -;R 1a is-CH 3 ; each R 1b is independently of the others hydrogen or-CH 3 ); Preferably, L 1 is 、 、 、 Or (b) ; And/or, each of said cycloalkyl, heterocyclyl and alkyl is independently unsubstituted; And/or, the atoms in the bispecific antibody drug conjugate are atoms in natural abundance.
  10. 10. The bispecific antibody drug conjugate of claim 8, wherein, L 1 is C 3 -C 6 saturated cycloalkyl or 3-6 membered saturated heterocyclyl, preferably L 1 is C 3 -C 6 saturated cycloalkyl, each of which is independently optionally substituted with one or more R 2a , R 2a is independently halogen or C 1 -C 6 alkyl; Preferably, L 1 is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl optionally substituted by one or more R 2a , R 2a each independently being halogen or C 1 -C 6 alkyl; Preferably, L 1 is 、 、 、 、 、 Or (b) 。
  11. 11. The bispecific antibody drug conjugate of claim 8, wherein in the structure of formula (A-1), M is-L 2 -L 1 -C (O) -; L 2 is-O-; L 1 is- (C (R 1a )(R 1b )) m -CH 2 -or C 3 -C 6 saturated cycloalkyl, said C 3 -C 6 saturated cycloalkyl optionally substituted with one or more R 2a ; m is selected from 1 or 2; R 1a and R 1b are each independently selected from hydrogen, halogen and C 1 -C 6 alkyl, said C 1 -C 6 alkyl optionally substituted with one or more halogens; Each R 2a is independently selected from halogen and C 1 -C 6 alkyl, said C 1 -C 6 alkyl optionally substituted with one or more halogen.
  12. 12. The bispecific antibody drug conjugate of claim 8, wherein the cytotoxic drug is selected from any one of the following structures: 、 、 、 、 、 、 、 、 、 、 And 。
  13. 13. The bispecific antibody drug conjugate of any one of claims 1-12, wherein the linker unit L is-L a -L b -L c -, and the L c is linked to the cytotoxic drug; -L a -is Or (b) ; -L b -is selected from any one of the following structures: 、 、 、 、 、 And Preferably Or (b) ; -L c -is 。
  14. 14. The bispecific antibody drug conjugate of claim 13, wherein the linker unit L is 、 Or (b) Preferably 。
  15. 15. The bispecific antibody drug conjugate of any one of claims 1-14, wherein the bispecific antibody drug conjugate is of the formula (a-2): ; Wherein p represents an average number of linkages and p is any integer or fraction from 1 to 10, preferably any integer or fraction from 2 to 8; ab is the bispecific antibody or antigen-binding fragment thereof of any one of claims 1-7; M is as defined in any one of claims 8 to 12; l is a linker unit L as claimed in claim 13 or 14.
  16. 16. The bispecific antibody drug conjugate of claim 15, wherein the bispecific antibody drug conjugate is of the formula (a-2 a) or (a-2 b): Or (b) ; Wherein, the P represents an average number of linkages and p is any integer or fraction from 1 to 10, preferably any integer or fraction from 2 to 8; ab is the bispecific antibody or antigen-binding fragment thereof of any one of claims 1-7; L 2 is-O-or-S-; preferably-O-; x 1 is C 3 -C 6 saturated cycloalkyl optionally substituted with 1,2 or 3R 2a ; X 2 is- (C (R 1a )(R 1b )) m -CH 2 -; m is 1 or 2; R 1a 、R 1b and R 2a are each independently hydrogen, halogen or C 1 -C 6 alkyl, said C 1 -C 6 alkyl optionally being substituted by one or more halogens.
  17. 17. The bispecific antibody drug conjugate of any one of claims 1-16, wherein the bispecific antibody drug conjugate is selected from any one of the following structures: 、 、 、 、 、 、 、 、 、 、 And ; Wherein p represents an average number of linkages and p is any integer or fraction from 1 to 10, preferably any integer or fraction from 2 to 8, preferably any integer or fraction from 4 to 8, preferably any integer or fraction from 6 to 8; ab is the bispecific antibody or antigen-binding fragment thereof of any one of claims 1-7.
  18. 18. A bispecific antibody drug conjugate, wherein the bispecific antibody drug conjugate is selected from any one of the following structures: 、 And ; Wherein p represents an average number of linkages and p is any integer or fraction from 1 to 10, preferably any integer or fraction from 3 to 8, preferably any integer or fraction from 4 to 8, preferably any integer or fraction from 6 to 8; DSYE001 is a bispecific antibody against B7H3 and PD-L1, the heavy chain amino acid sequence of which is shown as SEQ ID NO. 16, and the light chain amino acid sequence of which is shown as SEQ ID NO. 9.
  19. 19. A pharmaceutical composition comprising the bispecific antibody drug conjugate of any one of claims 1-18, and a pharmaceutically acceptable carrier or excipient.
  20. 20. Use of the bispecific antibody drug conjugate of any one of claims 1-19 or the pharmaceutical composition of claim 19 in the manufacture of a medicament for the treatment and/or prophylaxis of cancer, preferably, a B7H3 and/or PD-L1 positive expressed cancer.

Description

Bispecific antibody drug conjugate for resisting B7H3 and PD-L1, and preparation method and application thereof The application relates to a bispecific antibody drug conjugate with the application date of 2023, 12-27, the application number of 202380069557.8 and the application name of 'anti-B7H 3 and PD-L1', and a preparation method and application thereof. Technical Field The invention provides a bispecific antibody drug conjugate for resisting B7H3 and PD-L1, a preparation method and application thereof and a pharmaceutical composition containing the same. Background The B7-CD28 family, as a co-stimulatory signal for T lymphocyte activation, plays a critical role in the immune response in which T lymphocytes are involved. Studies have shown that different B7 molecular types have positive or negative regulation of immune cell responses. B7H3 (also known as CD 276) is a member of the B7 family, mainly expressed on the surface of tumor cells, chapoval AI et al have for the first time found that it has a costimulatory effect on cd4+ and cd8+ T cells, B7H3 signaling induces cellular immunity and selectively enhances interferon-gamma (IFN-gamma) production under T cell receptor signaling. However, as the research on B7H3 continues to be in progress, the inhibition function of B7H3 is gradually discovered, for example, it can inhibit proliferation of cd4+ T and cd8+ T cells. Furthermore, studies have shown that aberrant expression of B7H3 is associated with the occurrence, progression and metastasis of a variety of cancers, and there is a great deal of evidence that its high expression is associated with poor prognosis for various malignancies. B7H3 is highly expressed in all cancer types tested, has limited heterogeneity, and is rarely expressed in normal tissues. This suggests that B7H3 may be considered a Tumor Antigen (TA), providing the possibility for targeted therapies against B7H 3-highly expressing tumor cells. Currently, B7H3 targeted therapeutic strategies mainly include blocking monoclonal antibodies, radioimmunotherapy, antibody-drug conjugates (ADCs), monoclonal antibodies that mediate cytotoxicity, bispecific antibodies (BsAbs), and the like. Inhibitors targeting immune checkpoint PD1/PD-L1 are certainly the focus of tumor immunotherapy. PD-L1 is expressed on the surface of tumor cells, and a PD-L1 inhibitory antibody with cytotoxicity has better anti-tumor effect theoretically. However, only Avelumab of the marketed PD-L1 monoclonal antibodies were reported to mediate ADCC effects against tumors and to exhibit safety comparable to other PD-L1 antibodies. One important reason is that ADCC is due primarily to the abundance of antigen expression, whereas PD-L1 cannot be considered as a typical tumor antigen, and it has a large heterogeneity in tumor cells. PD1/PD-L1 pathway blocking antibodies are often used in combination with cytotoxic antibodies to enhance the effect of immunotherapy. The antibody drug conjugate (antibody drug conjugate, ADC) consists of three parts, an antibody or antigen binding fragment thereof (targeting), a linker and a small molecule drug. The antibody or antigen binding fragment thereof is conjugated with a small molecular drug such as cytotoxin having biological activity such as cytotoxicity through a cleavable or non-cleavable linker, and the specificity of targeting the antibody or antigen binding fragment thereof to a cell of interest (targeted cell) or the specificity of binding to a high-expression antigen and the high efficiency of the small molecular drug are fully utilized, so that the toxic and side effects on non-targeted cells are reduced or avoided. This means that antibody drug conjugates for tumors can precisely target tumor cells and reduce the impact on non-tumor cells compared to traditional tumor chemotherapeutic drugs. Among antibody-conjugated drugs targeting B7H3, the first three co-DS 7300 and MGC018 of MacroGenics progressed fastest, both drugs were currently in clinical phase II. MGC018 coupled DNA alkylating agent Du Kamei (duocarmycin) to humanized B7H3 antibody via a cleavable linker (linker). Early clinical trials showed that MGC018 exhibited preliminary anti-tumor activity and controlled toxicity in patients with advanced metastatic castration-resistant prostate cancer (mCRPC) and melanoma. DS7300 coupled irinotecan derivative (deruxtecan) to B7H3 antibody via a cleavable linker (linker), phase I clinical shows that anti-tumor activity was demonstrated in a variety of tumors, including mCRPC, small cell lung cancer, lung squamous carcinoma, esophageal squamous cell carcinoma, and endometrial carcinoma, and demonstrated superior safety. At present, the double antibody ADC is in an early clinical stage, and no medicines are available in the market. There remains a need in the art for bispecific antibody drug conjugates with synergistic mechanisms, better tumor selectivity and superior stability. Disclosure of Invention The invention aims to ove