Bispecific antibody against PD-1 and anti-VEGF, pharmaceutical composition thereof, and use thereof

Abstract

This application provides a bispecific antibody targeting PD-1 and VEGF, and a composition containing the same. Further, the application provides a nucleic acid molecule encoding the bispecific antibody, a vector and host cells for expressing the bispecific antibody, and therapeutic, diagnostic methods and uses of the antibody or its antigen-binding fragment.

Claims

1. A bispecific antibody comprising a first protein functional region and a second protein functional region, The first protein functional region is an anti-PD-1 antibody or its antigen-binding fragment, and the second protein functional region is an anti-VEGF antibody or its antigen-binding fragment, and the anti-VEGF antibody or its antigen-binding fragment includes a heavy chain variable region containing HCDR1, HCDR2, and HCDR3 whose amino acid sequences are shown in SEQ ID NO: 7, SEQ ID NO: 8, and SEQ ID NO: 9, respectively. Bispecific antibodies.

2. The anti-PD-1 antibody or its antigen-binding fragment includes a heavy chain variable region and / or a light chain variable region, and the amino acid sequences of HCDR1, HCDR2, and HCDR3 included in the heavy chain variable region are selected from the amino acid sequences of HCDR1, HCDR2, and HCDR3 included in the heavy chain variable region of nivolumab, pembrolizumab, semiprimab, tripalimab, cintilimab, or camrelizumab, respectively, and the amino acid sequences of LCDR1, LCDR2, and LCDR3 included in the light chain variable region are selected from the amino acid sequences of LCDR1, LCDR2, and LCDR3 included in the light chain variable region of nivolumab, pembrolizumab, semiprimab, tripalimab, cintilimab, or camrelizumab, respectively. The bispecific antibody according to claim 1.

3. The anti-PD-1 antibody or its antigen-binding fragment comprises a heavy chain variable region and / or a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3 whose amino acid sequences are shown in SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3 whose amino acid sequences are shown in SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively. A bispecific antibody according to claim 1 or 2.

4. The anti-PD-1 antibody or its antigen-binding fragment comprises a heavy chain variable region having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the amino acid sequence shown in SEQ ID NO: 10, and / or a light chain variable region having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the amino acid sequence shown in SEQ ID NO: 11, and / or The anti-VEGF antibody or its antigen-binding fragment includes a heavy chain variable region having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the amino acid sequence shown in SEQ ID NO:

12. A bispecific antibody according to any one of claims 1 to 3.

5. The anti-PD-1 antibody or its antigen-binding fragment is selected from Fab, Fab', F(ab')2, Fd, Fv, dAb, complementarity-determining region fragment, single-domain antibody, single-chain antibody, humanized antibody, chimeric antibody, or bi-antibody, and / or The anti-VEGF antibody or its antigen-binding fragment is selected from Fab, Fab', F(ab')2, Fd, Fv, dAb, complementarity-determining region fragment, single-domain antibody, single-chain antibody, humanized antibody, chimeric antibody, or bi-antibody. A bispecific antibody according to any one of claims 1 to 3.

6. The first protein functional region is an immunoglobulin, and / or The aforementioned second protein functional region is a single-domain antibody. A bispecific antibody according to any one of claims 1 to 5.

7. The immunoglobulin is selected from IgG, IgA, IgD, IgE, or IgM, and is preferably IgG. The bispecific antibody according to claim 6.

8. The constant region of the immunoglobulin is derived from a human antibody. Preferably, the constant region of the immunoglobulin is selected from the constant regions of human IgG1, IgG2, IgG3, or IgG4. The bispecific antibody according to claim 6.

9. There are two of the single-domain antibodies, and one end of each single-domain antibody is ligated to the C-terminus or N-terminus of the two heavy chains of the immunoglobulin, or One end of each single-domain antibody is ligated to the C-terminus or N-terminus of the two light chains of the immunoglobulin, or The N-terminus of each single-domain antibody is ligated to the C-terminus of the heavy chain constant region CH1 of the immunoglobulin, and the C-terminus of each single-domain antibody is ligated to the N-terminus of the heavy chain constant region CH2 of the immunoglobulin. The bispecific antibody according to claim 6.

10. (1) The first polypeptide chain of the bispecific antibody is represented by the formula VLPD-1-CL, and the second polypeptide chain is represented by the formula VHVEGF-linker-VHPD-1-CH1-hinge-CH2-CH3, or (2) The first polypeptide chain of the bispecific antibody is represented by the formula VLPD-1-CL, and the second polypeptide chain is represented by the formula VHPD-1-CH1-hinge-CH2-CH3-linker-VHVEGF, or (3) The first polypeptide chain of the bispecific antibody is represented by the formula VLPD-1-CL, and the second polypeptide chain is represented by the formula VHPD-1-CH1-linker-VHVEGF-linker-hinge-CH2-CH3, or (4) The first polypeptide chain of the bispecific antibody is represented by the formula VHVEGF-linker-VLPD-1-CL, and the second polypeptide chain is represented by the formula VHPD-1-CH1-hinge-CH2-CH3. The bispecific antibody according to claim 9.

11. (1) The first polypeptide chain comprises the amino acid sequence shown in SEQ ID NO: 13, and the second polypeptide chain comprises the amino acid sequence shown in SEQ ID NO: 14, or (2) The first polypeptide chain comprises the amino acid sequence shown in SEQ ID NO: 13, and the second polypeptide chain comprises the amino acid sequence shown in SEQ ID NO: 15, or (3) The first polypeptide chain comprises the amino acid sequence shown in SEQ ID NO: 13, and the second polypeptide chain comprises the amino acid sequence shown in SEQ ID NO: 16, or (4) The first polypeptide chain comprises the amino acid sequence shown in SEQ ID NO: 17, and the second polypeptide chain comprises the amino acid sequence shown in SEQ ID NO:

18. The bispecific antibody according to claim 10.

12. Encoding a bispecific antibody according to any one of claims 1 to 11, Isolated nucleic acid molecules.

13. A comprising the isolated nucleic acid molecule described in claim 12, vector.

14. The isolated nucleic acid molecule according to claim 12, or the vector according to claim 13, host cell.

15. A method for producing a bispecific antibody according to any one of claims 1 to 11, comprising the steps of culturing a host cell according to claim 14 and obtaining the bispecific antibody from the culture, method.

16. A conjugate comprising a bispecific antibody and a conjugation moiety according to any one of claims 1 to 11, wherein the conjugation moiety is a therapeutically active substance or a detectable label, preferably the therapeutically active substance is a cytotoxin, cytokine or radionuclide, and preferably the detectable label is selected from a radioisotope, fluorescent substance, luminescent substance, colored substance or enzyme. Conjugate.

17. A reagent kit comprising a bispecific antibody according to any one of claims 1 to 11, or a conjugate according to claim 16, Preferably, the reagent kit further comprises a second antibody capable of specifically binding to the bispecific antibody, preferably the second antibody further comprises a detectable label, and more preferably the detectable label is selected from radioisotopes, fluorescent substances, luminescent substances, colored substances, or enzymes. Reagent kit.

18. A pharmaceutical composition comprising a bispecific antibody according to any one of claims 1 to 11, or a conjugate according to claim 16, preferably further comprising a pharmaceutically acceptable adjuvant. Pharmaceutical composition.

19. A method for treating or improving a malignant tumor, comprising administering to a patient in need a bispecific antibody according to any one of claims 1 to 11, a conjugate according to claim 16, or a pharmaceutical composition according to claim 18, wherein the malignant tumor is a tumor overexpressing PD-L1 and / or VEGF, preferably a solid tumor, more preferably lung cancer, colon cancer, colorectal cancer, melanoma, or ovarian cancer, more preferably lung cancer is non-small cell lung cancer or small cell lung cancer, more preferably non-small cell lung cancer is squamous cell non-small cell lung cancer or non-squamous cell non-small cell lung cancer, and more preferably non-squamous cell non-small cell lung cancer is locally advanced or metastatic non-squamous cell non-small cell lung cancer. method.

20. A method for regulating the immune response and tumor angiogenesis, comprising administering a bispecific antibody according to any one of claims 1 to 11, a conjugate according to claim 16, or a pharmaceutical composition according to claim 18 to a patient in need, wherein the regulation of the immune response is achieved by blocking the binding of PD-1 to its receptor, and the regulation of tumor angiogenesis is achieved by blocking the binding of VEGF to its receptor. method.

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