阅读说明：本技术 抗pd-l1抗体及其应用 (anti-PD-L1 antibody and application thereof ) 是由 房健民 姜静 李慎军 赵国锐 于 2020-08-25 设计创作，主要内容包括：本发明提供了一种PD-L1免疫抑制剂,具体提供了一种能够靶向PD-L1的单克隆抗体以及编码这些抗体的核苷酸、多核苷酸组合、表达载体以及表达载体组合。本发明还提供了包含上述抗PD-L1抗体的偶联物或者药物组合物。本发明还提供了上述抗PD-L1抗体核苷酸、多核苷酸组合、表达载体、表达载体组合、偶联物或者药物组合物在治疗或预防癌症的药物中的用途。(The invention provides a PD-L1 immunosuppressant, and particularly provides a monoclonal antibody capable of targeting PD-L1, and nucleotides, polynucleotide combinations, expression vectors and expression vector combinations for encoding the antibodies. The invention also provides a conjugate or a pharmaceutical composition comprising the anti-PD-L1 antibody. The invention also provides application of the anti-PD-L1 antibody nucleotide, the polynucleotide combination, the expression vector combination, the conjugate or the pharmaceutical composition in preparing a medicament for treating or preventing cancer.)

An isolated anti-PD-L1 antibody, or antigen-binding fragment thereof, comprising a heavy chain variable region and a light chain variable region, the CDRs of the heavy chain variable region and/or the CDRs of the light chain variable region having the same CDR sequences or having 1-2 amino acid substitutions in the CDRs of an antibody defined by the sequences:

(1) the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 31; and/or

(2) The amino acid sequence of the light chain variable region is shown as SEQ ID NO: 32.

The antibody or antigen-binding fragment thereof of claim 1, wherein:

(1) the heavy chain variable region CDR1 has the amino acid sequence shown in SEQ ID NO. 1, 7, 13, 19 or 25 or the amino acid sequence obtained by 1 or 2 amino acid substitutions of SEQ ID NO. 1, 7, 13, 19 or 25; CDR2 amino acid sequence is shown in SEQ ID NO. 2, 8, 14, 20 or 26 or the amino acid sequence after 1 or 2 amino acid substitutions are carried out on SEQ ID NO. 2, 8, 14, 20 or 26; CDR3 amino acid sequence is shown in SEQ ID NO. 3, 9, 15, 21 or 27 or amino acid sequence obtained by 1 or 2 amino acid substitutions of SEQ ID NO. 3, 9, 15, 21 or 27; and/or

(2) The light chain variable region CDR1 has the amino acid sequence shown in SEQ ID NO. 4, 10, 16, 22 or 28 or the amino acid sequence obtained by 1 or 2 amino acid substitutions of SEQ ID NO. 4, 10, 16, 22 or 28; CDR2 amino acid sequence is shown in SEQ ID NO. 5, 11, 17, 23 or 29 or amino acid sequence obtained by 1 or 2 amino acid substitutions of SEQ ID NO. 5, 11, 17, 23 or 29; the CDR3 amino acid sequence is shown in SEQ ID NO. 6, 12, 18, 24 or 30 or the amino acid sequence obtained by 1 or 2 amino acid substitutions of SEQ ID NO. 6, 12, 18, 24 or 30.

The antibody or antigen-binding fragment thereof according to claim 2, wherein,

(1) the heavy chain variable region CDR1-3 amino acid sequence is SEQ ID NO:1-3 or 1 or 2 amino acid substitutions to SEQ ID NO 1-3; and/or the amino acid sequence of CDR1-3 in the variable region of the light chain is SEQ ID NO: 4-6 or to SEQ ID NO: 4-6 amino acid sequence after 1 or 2 amino acid substitutions; or

(2) The heavy chain variable region CDR1-3 amino acid sequence is SEQ ID NO:7-9 or an amino acid sequence obtained by 1 or 2 amino acid substitutions of SEQ ID NO 7-9; and/or the amino acid sequence of CDR1-3 in the variable region of the light chain is SEQ ID NO: 10-12 or to SEQ ID NO: 10-12 amino acid sequence after 1 or 2 amino acid substitutions; or

(3) The heavy chain variable region CDR1-3 amino acid sequence is SEQ ID NO:13-15 or an amino acid sequence obtained by 1 or 2 amino acid substitutions of SEQ ID NO 13-15; and/or the amino acid sequence of CDR1-3 in the variable region of the light chain is SEQ ID NO: 16-18 or to SEQ ID NO: 16-18 by 1 or 2 amino acid substitutions; or

(4) The heavy chain variable region CDR1-3 amino acid sequence is SEQ ID NO:19-21 or an amino acid sequence obtained by substituting 1 or 2 amino acids for SEQ ID NO 19-21; and/or the amino acid sequence of CDR1-3 in the variable region of the light chain is SEQ ID NO: 22-24 or to SEQ ID NO: 22-24 by 1 or 2 amino acid substitutions; or

(5) The heavy chain variable region CDR1-3 amino acid sequence is SEQ ID NO:25-27 or an amino acid sequence obtained by substituting 1 or 2 amino acids for SEQ ID NO 25-27; and/or the light chain variable region CDR1-3 amino acid sequence is SEQ ID NO: 28-30 or to SEQ ID NO: 28-30 by 1 or 2 amino acid substitutions.

The antibody or antigen-binding fragment thereof of any one of claims 1-3, wherein the heavy chain variable region CDR1-3 amino acid sequence is SEQ ID NO: 1-3; and/or the amino acid sequence of CDR1-3 in the variable region of the light chain is SEQ ID NO: 4-6.

The antibody or antigen-binding fragment thereof of any one of claims 1-4, wherein:

(1) the heavy chain variable region has the sequence shown as SEQ ID NO. 31 or the sequence with the same CDR1-3 as SEQ ID NO. 31 and the identity of more than 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% compared with SEQ ID NO. 31; and/or

(2) The light chain variable region has the sequence shown in SEQ ID NO. 32 or the sequence with the same CDR1-3 as SEQ ID NO. 32 and the identity greater than 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% as compared with SEQ ID NO. 32.

The antibody or antigen-binding fragment thereof of claim 5, wherein:

(1) the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO: 31; and/or

(2) The amino acid sequences of the light chain variable regions are shown in SEQ ID NO:32, respectively.

The antibody or antigen-binding fragment thereof of claim 6, wherein:

(1) the amino acid sequence of the heavy chain is shown as SEQ ID NO. 33; and/or

(2) The amino acid sequences of the light chains are shown in SEQ ID NO 34, respectively.

The antibody or antigen binding fragment thereof of any one of claims 1-7, wherein the antibody is a monoclonal antibody, a chimeric antibody, a humanized antibody, a bispecific antibody, a multispecific antibody or Fab fragment, a F (ab')₂Fragments, Fv fragments, dAbs, Fd, single chain antibodies (scFv).

The antibody or antigen binding fragment thereof of any one of claims 1-8, wherein the antibody is a humanized monoclonal antibody.

The antibody or antigen-binding fragment thereof of any one of claims 1-9, further comprising a human or murine constant region.

The antibody or antigen binding fragment thereof of claim 10, wherein said constant region is selected from the group consisting of IgG1, IgG2, IgG3, IgG 4.

The antibody or antigen binding fragment thereof of claim 11, wherein the constant region is IgG1, IgG2A, IgG 2B.

An isolated polynucleotide encoding the antibody or antigen-binding fragment thereof of any one of claims 1-12.

A combination of isolated polynucleotides comprising: a polynucleotide encoding the heavy chain of the antibody or antigen-binding fragment thereof of any one of claims 1-12 and a polynucleotide encoding the light chain of the antibody or antigen-binding fragment thereof of any one of claims 1-12.

A nucleic acid construct comprising a polynucleotide according to claim 13.

The nucleic acid construct of claim 15, which is a vector.

A host cell comprising the nucleic acid construct of claim 15 or the vector of claim 16.

The host cell of claim 17, which is a prokaryotic cell, a eukaryotic cell, a yeast cell, a mammalian cell, an e.coli cell or a CHO cell, an NS0 cell, an Sp2/0 cell, a BHK cell.

An antibody drug conjugate comprising the antibody or antigen-binding portion thereof of any one of claims 1-12 and a drug toxin.

The antibody drug conjugate of claim 19, wherein the drug toxin is selected from the group consisting of chemical substances, toxins, polypeptides, enzymes, isotopes, cytokines, antibodies or other biologically active substances or mixtures thereof that directly or indirectly inhibit cell growth or kill cells, or inhibit or kill cells by activating immune response in the body, thereby treating tumors, preferably interleukins, tumor necrosis factors, chemokines, nanoparticles, MMAE, MMAF, DM1, DM4, Calicheamicin (Calicheamicin), duocarmycin, and Doxorubicin (Doxorubicin).

A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof of any one of claims 1-12 and/or the conjugate of claim 19 or 20, and a pharmaceutically acceptable carrier.

A method of making an anti-PD-L1 antibody, comprising culturing the host cell of claim 17 or 18 under conditions suitable for expression of a vector encoding an anti-PD-L1 antibody or antigen-binding fragment, and recovering the antibody or fragment.

A method of enhancing T cell function comprising administering to a dysfunctional T cell an effective amount of the pharmaceutical composition of claim 21.

A method of treating a disorder of T cell dysfunction, comprising administering to a patient having a disorder of T cell dysfunction, including infection, tumor immunity, a therapeutically effective amount of the pharmaceutical composition of claim 21.

The method of claim 24, wherein said tumor immunity results from a cancer selected from the group consisting of: breast, lung, colon, ovarian, melanoma, bladder, kidney, liver, salivary gland, stomach, glioma, thyroid, thymus, epithelial, head and neck, stomach and pancreatic cancer.

Use of the antibody or antigen-binding fragment thereof of any one of claims 1-12, the polynucleotide of claim 13, the polynucleotide combination of claim 14, the nucleic acid construct of claim 15, the vector of claim 16, the antibody drug conjugate of claim 19 or 20, or the pharmaceutical composition of claim 21 in the manufacture of a medicament for the treatment or prevention of cancer.

The use of claim 26, wherein the cancer is a solid tumor.

The use of claim 27, wherein the solid tumor is lung cancer, colorectal cancer, breast cancer, ovarian cancer, melanoma, bladder cancer, urothelial cancer, kidney cancer, liver cancer, salivary gland cancer, stomach cancer, glioma, thyroid cancer, thymus cancer, epithelial cancer, head and neck cancer, stomach cancer, pancreatic cancer, Merkel cell carcinoma.

The use of claim 28, wherein the lung cancer is non-small cell lung cancer.

The use of claim 29, wherein the ovarian cancer is triple negative breast cancer.