Pharmaceutical composition and use

Abstract

A pharmaceutical composition comprising an anti-LAG3-anti-PD-1 bispecific antibody and an anti-CD47 monoclonal antibody and a use of the pharmaceutical composition in the preparation of a drug for treating or preventing tumors.

Description

As used herein, when referring to the amino acid sequence of PD-1 protein (NCBI GenBank: NM_005018), it includes the full length of PD-1 protein, or the extracellular fragment PD-1 ECD of PD-1, or a fragment comprising PD-1 ECD, and it also includes a fusion protein of the full length of PD-1 protein or a fusion protein of PD-1 ECD, such as a fragment fused to an Fc protein fragment of mouse or human IgG (mFc or hFc). However, those skilled in the art will appreciate that in the amino acid sequence of the PD-1 protein, mutations or variations (including but not limited to, substitutions, deletions, and / or additions) can be naturally produced or artificially introduced without affecting biological functions thereof. Therefore, in the present disclosure, the term “PD-1 protein” shall include all such sequences, including their natural or artificial variants. In addition, when describing a sequence fragment of the PD-1 protein, it also includes the corresponding sequence fragments in their natural or artificial variants. As used herein, when referring to the amino acid sequence of lymphocyte-activation gene 3 (LAG3), it includes the full length of the LAG3 protein, or the extracellular fragment LAG3 ECD of LAG3, or a fragment comprising LAG3 ECD, and it also includes a fusion protein of the full length of the LAG3 protein or a fusion protein of LAG3 ECD, such as a fragment fused to an Fc protein fragment of mouse or human IgG (mFc or hFc). However, those skilled in the art will appreciate that in the amino acid sequence of the LAG3 protein, mutations or variations (including but not limited to, substitutions, deletions, and / or additions) can be naturally produced or artificially introduced without affecting biological functions thereof. Therefore, in the present disclosure, the term “LAG3 protein” shall include all such sequences, including their natural or artificial variants. In addition, when describing a sequence fragment of the LAG3 protein, it also includes the corresponding sequence fragments in their natural or artificial variants. As used herein, when referring to the amino acid sequence of a CD47 protein (NCBI GenBank: NP_001768.1), it includes the full length of the CD47 protein, or the extracellular fragment CD47 ECD of CD47, or a fragment comprising CD47 ECD, and it also includes a fusion protein of the full length of the CD47 protein or a fusion protein of CD47 ECD, such as a fragment fused to an Fc protein fragment of mouse or human IgG (mFc or hFc). However, those skilled in the art will appreciate that in the amino acid sequence of the CD47 protein, mutations or variations (including but not limited to, substitutions, deletions, and / or additions) can be naturally produced or artificially introduced without affecting biological functions thereof. Therefore, in the present disclosure, the term “CD47 protein” shall include all such sequences, including their natural or artificial variants. In addition, when the sequence fragment of the CD47 protein is described, the CD47 protein also includes the corresponding sequence fragments in natural or artificial variants thereof. In the present disclosure, the term “ADCP” refers to antibodydependent cellular phagocytosis. The Fc fragment of an antibody that binds to a cell surface antigen binds to the Fc receptor of a phagocytically active cell, such as a macrophage, which in turn mediates phagocytosis of the antibody-bound cells by phagocytic cells. In the present disclosure, the terms “first” (e.g., first protein functional region, first antibody, or first linker), “second” (e.g., second protein functional region, second antibody, or second linker), “third” (e.g., third linker), and “fourth” (e.g., fourth linker) are used for distinguishing or clarity in expression and do not carry typical sequential meanings, unless otherwise specified. The contents of prior applications CN202211199026.4 (publication No. CN 115873123A) and CN202010919545.8 (publication No. CN112442123A) are incorporated herein by reference in their entirety. Beneficial Effects of the Present Disclosure The present disclosure achieves one or more of the following effects: (1) The therapeutic combination of the present disclosure is capable of effectively preventing or treating a tumor. (2) The combination of the anti-LAG3-anti-PD-1 bispecific antibody and the anti-CD47 monoclonal antibody has a pharmacological effect of effectively inhibiting tumors, which is superior to that of either the anti-LAG3-anti-PD-1 bispecific antibody or the anti-CD47 monoclonal antibody alone, achieving a synergistic technical effect. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 shows phagocytosis of tumor cells by macrophages promoted by administration of the anti-LAG3-anti-PD-1 bispecific antibody in combination with the anti-CD47 monoclonal antibody. Hybridoma cell line LT012 was deposited at China Center for Type Culture Collection (CCTCC) on June 21, 2018 with a CCTCC designation of CCTCC NO. C2018135, the depository address being Wuhan University, Wuhan, China, postal code: 430072. DETAILED DESCRIPTION The embodiments of the present disclosure will be described in detail below with reference to the examples. Those skilled in the art will appreciate that the following examples are only for illustrating the present disclosure, and should not be construed as limitations to the scope of the present disclosure. Examples where the specific technologies or conditions are not specified are performed according to the technologies or conditions described in the publications of the art (e.g., see Molecular Cloning: A Laboratory Manual, authored by J. Sambrook et al., and translated by Huang Peitang et al., third edition, Science Press) or according to the package insert. Reagents or instruments used are commercially available conventional products if the manufacturers thereof are not specified. In the examples of the present disclosure, the isotype control antibody used, i.e., hIgG4, was an antibody targeting human anti-hen egg lysozyme (HEL), and the variable region sequence of the antibody is from the study reported by Acierno et al., entitled “Affinity maturation increases the stability and plasticity of the Fv domain of anti-protein antibodies” (Acierno et al., J Mol Biol., 2007; 374(1): 130-46). hIgG4 employs Ig gamma-4 chain C region (ACCESSION: P01861.1) as the heavy chain constant region, with the S228P mutation introduced to enhance stability, and employs Ig kappa chain C region (ACCESSION: P01834) as the light chain constant region. The hIgG4 is prepared in the laboratory of Akeso Biopharma Inc. Preparation Example 1: Design and Preparation of Anti-LAG3 Antibodies 1. Design of antibodies The inventors creatively designed a series of antibody sequences based on the known LAG3 protein sequence (NCBI Reference Sequence: NP_002277.4), the three-dimensional crystal structure thereof, etc. Through extensive screening and testing, humanized monoclonal antibodies specifically binding to LAG3 were finally obtained, named H7L8, H7L9 and H7L10, respectively. The amino acid sequences of the heavy and light chain variable regions of the monoclonal antibodies and the encoding sequences thereof are as follows. Nucleotide sequence of the heavy chain variable region H7v of H7L8 (360 bp): CAGGTGCAGCTGCAGCAGTGGGGAGCTGGACTGCTGAAA CCTAGCGAGACACTGAGCCTGACCTGTGCTGTGTACGGCGGA TCTATCAGCGATTACTACTGGAACTGGATCAGGCAGCCCCCTG GAAAGGGACTGGAATGGATCGGAGAGATCAACCACAGGGGCA CCACCAACTCCAATCCCTCTCTGAAGAGCAGGGTGACACTGA GCCTCGACACAAGCAAGAATCAGTTCAGCCTGAAGCTGAGGT CCGTGACCGCTGCTGATACAGCTGTGTACTACTGTGCCTTCGG CTACAGCGATTACGAGTACGATTGGTTCGACCCTTGGGGCCA GGGAACACTGGTTACAGTGAGCTCC (SEQ ID NO: 1) Amino acid sequence of the heavy chain variable region H7v of H7L8 (120 aa): QVQLQQWGAGLLKPSETLSLTCAVYGGSISDYYWNWIRQPP GKGLEWIGEINHRGTTNSNPSLKSRVTLSLDTSKNQFSLKLRSVT AADTAVYYCAFGYSDYEYDWFDPWGQGTLVTVSS (SEQ ID NO: 2) Nucleotide sequence of the light chain variable region L8v of H7L8 (321 bp): GAGATCGTTCTGACCCAGAGCCCAGCTACACTGAGCCTGT CTCCTGGAGAGAGGGCTACACTGTCCTGCAGAGCTAGCCAGA CCATCAGCAGCTACCTGGCTTGGTACCAGCAGAAGCCTGGCC AAGCTCCAAGGCTGCTGATCTACGACGCCTCTAATAGGGCCA CCGGCATCCCTGCTAGATTCTCTGGAAGCGGCAGCGGAACCG ACTTTACACTGACAATCAGCTCCCTGGAGCCCGAGGATTTCGC TGTTTACTACTGTCAGCAGCGCAGCAACTGGCCCATCACATTC GGACAGGGCACAAATCTGGAGATCAAG (SEQ ID NO: 3) Amino acid sequence of the light chain variable region L8v of H7L8 (107 aa): EIVLTQSPATLSLSPGERATLSCRASQTISSYLAWYQQKPGQA PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQ QRSNWPITFGQGTNLEIK (SEQID NO: 4) The nucleotide sequence of the heavy chain variable region H7v of H7L9 is identical to the nucleotide sequence of the heavy chain variable region H7v of H7L8, as set forth in SEQ ID NO: 1. The amino acid sequence of the heavy chain variable region H7v of H7L9 is identical to the amino acid sequence of the heavy chain variable region H7v of H7L8, as set forth in SEQ ID NO: 2. Nucleotide sequence of the light chain variable region L9v of H7L9 (321 bp): GAGATCGTTCTGACCCAGAGCCCAGCTACACTGAGCCTGT CTCCTGGAGAGAGGGCTACACTGTCCTGCAGAGCTAGCCAGA CCATCAGCAGCTACCTGGCTTGGTACCAGCAGAAGCCTGGCC AAGCTCCAAGGCTGCTGATCTACGACGGCTCTAATAGGGCCA CCGGCATCCCTGCTAGATTCTCTGGAAGCGGCAGCGGAACCG ACTTTACACTGACAATCAGCTCCCTGGAGCCCGAGGATTTCGC TGTTTACTACTGTCAGCAGCGCAGCAACTGGCCCCTCACATTC GGACAGGGCACAAATCTGGAGATCAAG (SEQ ID NO: 41) Amino acid sequence of the light chain variable region L9v of H7L9 (107 bp): EIVLTQSPATLSLSPGERATLSCRASQTISSYLAWYQQKPGQA PRLLIYDGSNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQ QRSNWPLTFGQGTNLEIK (SEQ ID NO: 42) The nucleotide sequence of the heavy chain variable region H7v of H7L10 is identical to the nucleotide sequence of the heavy chain variable region H7v of H7L8, as set forth in SEQ ID NO: 1. The amino acid sequence of the heavy chain variable region H7v of H7L10 is identical to the amino acid sequence of the heavy chain variable region H7v of H7L8, as set forth in SEQ ID NO: 2. Nucleotide sequence of the light chain variable region L10v of H7L10 (321 bp): GAGATCGTTCTGACCCAGAGCCCAGCTACACTGAGCCTGT CTCCTGGAGAGAGGGCTACACTGTCCTGCAGAGCTAGCCAGT CCATCAGCAGCTACCTGGCTTGGTACCAGCAGAAGCCTGGCC AAGCTCCAAGGCTGCTGATCTACGACGGCTCTAATAGGGCCA CCGGCATCCCTGCTAGATTCTCTGGAAGCGGCAGCGGAACCG ACTTTACACTGACAATCAGCTCCCTGGAGCCCGAGGATTTCGC TGTTTACTACTGTCAGCAGCGCAGCAACTGGCCCATCACATTC GGACAGGGCACAAATCTGGAGATCAAG (SEQ ID NO: 43) Amino acid sequence of the light chain variable region L10v of H7L10 (107 bp): EIVLTQSPATLSLSPGERATLSCRASQSISSYLAWYQQKPGQA PRLLIYDGSNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQ QRSNWPITFGQGTNLEIK (SEQ ID NO: 44) The amino acid sequences of the CDRs of the antibody H7L8 are as follows (according to the IMGT numbering system): HCDR1: GGSISDYY (SEQ ID NO: 5); HCDR2: INHRGTT (SEQ ID NO: 6); HCDR3: AFGYSDYEYDWFDP (SEQ ID NO: 7); LCDR1: QTISSY (SEQ ID NO: 8); LCDR2: DAS (SEQ ID NO: 9); LCDR3: QQRSNWPIT (SEQ ID NO: 10). The amino acid sequences of the CDRs of the antibody H7L9 are as follows (according to the IMGT numbering system): HCDR1: GGSISDYY (SEQ ID NO: 5); HCDR2: INHRGTT (SEQ ID NO: 6); HCDR3: AFGYSDYEYDWFDP (SEQ ID NO: 7); LCDR1: QTISSY (SEQ ID NO: 8); LCDR2: DGS (SEQ ID NO: 46); LCDR3: QQRSNWPLT (SEQ ID NO: 47). The amino acid sequences of the CDRs of the antibody H7L10 are as follows (according to the IMGT numbering system): HCDR1: GGSISDYY (SEQ ID NO: 5); HCDR2: INHRGTT (SEQ ID NO: 6); HCDR3: AFGYSDYEYDWFDP (SEQ ID NO: 7); LCDR1: QSISSY (SEQ ID NO: 48); LCDR2: DGS (SEQ ID NO: 46); LCDR3: QQRSNWPIT (SEQ ID NO: 10). 2. Expression and purification of humanized antibody H7L8(hG1WT) The heavy chain cDNA sequence (the encoding sequence of the variable region is set forth in SEQ ID NO: 1; the constant region is Ig gamma-1 chain C region, SEQ ID NO: 39) and the light chain cDNA sequence (the encoding sequence of the variable region is set forth in SEQ ID NO: 3; the constant region is P01834.1 (human Ig kappa chain C region, SEQ ID NO: 40) of H7L8(hG1WT) were separately cloned into pUC57simple vectors (supplied by GenScript), and plasmids pUC57simple-H7 and pUC57simple-L8 were obtained, respectively. The plasmids pUC57simple-H7 and pUC57simple-L8 were each digested (HindIII&EcoRI). The heavy and light chains isolated by electrophoresis were separately subcloned into pcDNA3.1 vectors, and recombinant plasmids were extracted to co-transfect 293F cells. After 7 days of cell culture, the culture solution was separated by high-speed centrifugation, and the supernatant was concentrated and loaded onto a HiTrap MabSelect SuRe column. The protein was eluted in one step with an elution buffer. The target sample was isolated and buffer-exchanged into PBS. Amino acid sequence of the heavy chain constant region of H7L8(hG1WT) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK PSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPGK (SEQ ID NO: 39) Amino acid sequence of the light chain constant region of H7L8(hG1WT) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWK VDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 40) 3. Design of humanized antibody H7L8(hG1TM) On the basis of H7L8(hG1WT), the inventors obtained a humanized antibody H7L8(hG1TM) with constant region mutations by introducing a leucine-to-alanine point mutation at position 234 (according to the EU numbering system, the same below) (L234A), a leucine-to-alanine point mutation at position 235 (L235A), and a glycine-to-alanine point mutation at position 237 (G237A) in the heavy chain. The amino acid sequence of the heavy chain H7(hG1TM) of H7L8(hG1TM) is set forth in SEQ ID NO: 11, and the amino acid sequence of the light chain L8 thereof is set forth in SEQ ID NO: 12. The humanized antibody H7L8(hG1TM) was prepared by the method described above in step 2. Amino acid sequence of the heavy chain H7(hG1TM) of H7L8(hG1TM) QVQLQQWGAGLLKPSETLSLTCAVYGGSISDYYWNWIRQPP GKGLEWIGEINHRGTTNSNPSLKSRVTLSLDTSKNQFSLKLRSVT AADTAVYYCAFGYSDYEYDWFDPWGQGTLVTVSSASTKGPSVFP LAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEP KSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPGK (SEQ ID NO: 11) Amino acid sequence of the light chain L8 of H7L8(hG1TM) EIVLTQSPATLSLSPGERATLSCRASQTISSYLAWYQQKPGQA PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQ QRSNWPITFGQGTNLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCL LNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 12) 4. Design of humanized antibody H7L8(hG4DM) On the basis of H7L8(hG1WT), the inventors obtained a humanized antibody H7L8(hG4DM) with constant region mutations by using Ig gamma-4 chain C region as a heavy chain constant region and introducing a phenylalanine-to-alanine point mutation at position 234 (F234A) and a leucine-to-alanine point mutation at position 235 (L235A) in the heavy chain constant region while keeping the antibody variable region unchanged. The amino acid sequence of the heavy chain of H7L8(hG4DM) is set forth in SEQ ID NO: 13, and the amino acid sequence of the light chain thereof is set forth in SEQ ID NO: 12. Amino acid sequence of the heavy chain H7(hG4DM) of H7L8(hG4DM): QVQLQQWGAGLLKPSETLSLTCAVYGGSISDYYWNWIRQPP GKGLEWIGEINHRGTTNSNPSLKSRVTLSLDTSKNQFSLKLRSVT AADTAVYYCAFGYSDYEYDWFDPWGQGTLVTVSSASTKGPSVFP LAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVES KYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVD VSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVL HQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPS QEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS LSLGK (SEQ ID NO: 13) The amino acid sequence of the light chain L8 of H7L8(hG4DM) is identical to the amino acid sequence of the light chain of H7L8(hG1TM), as set forth in SEQ ID NO: 12. 5. Expression and purification of humanized antibodies H7L8(hG4WT), H7L9(hG4WT) and H7L10(hG4WT) The heavy chain cDNA sequences (the encoding sequences of the variable regions are set forth in SEQ ID NO: 1; the constant regions are Ig gamma-4 chain C regions, set forth in SEQ ID NO: 45) of H7L8(hG4WT), H7L9(hG4WT) and H7L10(hG4WT), the light chain cDNA sequence (the encoding sequence of the variable region is set forth in SEQ ID NO: 3; the constant region is human Ig kappa chain C region, set forth in SEQ ID NO: 40) of H7L8(hG4WT), the light chain cDNA sequence (the encoding sequence of the variable region is set forth in SEQ ID NO: 42; the constant region is human Ig kappa chain C region, set forth in SEQ ID NO: 40) of H7L9(hG4WT), and the light chain cDNA sequence (the encoding sequence of the variable region is set forth in SEQ ID NO: 44; the constant region is human Ig kappa chain C region, set forth in SEQ ID NO: 40) of H7L10(hG4WT) were separately cloned into pUC57simple vectors (supplied by GenScript), and plasmids pUC57simple-H7, pUC57simple-L8, pUC57simple-L9 and pUC57simple-L10 were obtained, respectively. The plasmids pUC57simple-H7, pUC57simple-L8, pUC57simple-L9 and pUC57simple-L10 were each digested (HindIII&EcoRI). The heavy and light chains isolated by electrophoresis were separately subcloned into pcDNA3.1 vectors, and recombinant plasmids were extracted to co-transfect 293F cells. After 7 days of cell culture, the culture solution was separated by high-speed centrifugation, and the supernatant was concentrated and loaded onto a HiTrap MabSelect SuRe column. The protein was eluted in one step with an elution buffer. The target sample was isolated and buffer-exchanged into PBS. Amino acid sequence of the heavy chain constant region of H7L8(hG4WT), H7L9(hG4WT), or H7L10(hG4WT): ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNS GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHK PSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMIS RTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQ PREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEA LHNHYTQKSLSLSLGK (SEQ ID NO: 45) Amino acid sequence of the light chain constant region of H7L8(hG4WT), H7L9(hG4WT), or H7L10(hG4WT): RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWK VDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 40) Preparation Example 2: Design and Preparation of Anti-PD-1 Antibody 14C12 and Its Humanized Antibody 14C12H1L1 The amino acid sequences and encoding nucleotide sequences of the heavy and light chains of the anti-PD-1 antibody 14C12 and its humanized antibody 14C12H1L1 are identical to those of 14C12 and 14C12H1L1 in Chinese Patent Publication No. CN 106967172A (or No. CN 106977602A), respectively. (1) Heavy and light chain variable region sequences of 14C12 Nucleotide sequence of the heavy chain variable region of 14C12: (354 bp) GAGGTCAAACTGGTGGAGAGCGGCGGCGGGCTGGTGAAG CCCGGCGGGTCACTGAAACTGAGCTGCGCCGCTTCCGGCTTC GCCTTTAGCTCCTACGACATGTCATGGGTGAGGCAGACCCCT GAGAAGCGCCTGGAATGGGTCGCTACTATCAGCGGAGGCGGG CGATACACCTACTATCCTGACTCTGTCAAAGGGAGATTCACAA TTAGTCGGGATAACGCCAGAAATACTCTGTATCTGCAGATGTC TAGTCTGCGGTCCGAGGATACAGCTCTGTACTATTGTGCAAAC CGGTACGGCGAAGCATGGTTTGCCTATTGGGGACAGGGCACC CTGGTGACAGTCTCTGCC (SEQ ID NO: 14) Amino acid sequence of the heavy chain variable region of 14C12: (118 aa) EVKLVESGGGLVKPGGSLKLSCAASGFAFSSYDMSWVRQTP EKRLEWVATISGGGRYTYYPDSVKGRFTISRDNARNTLYLQMSS LRSEDTALYYCANRYGEAWFAYWGQGTLVTVSA (SEQ ID NO: 15) Nucleotide sequence of the light chain variable region of 14C12: (321 bp) GACATTAAGATGACACAGTCCCCTTCCTCAATGTACGCTA GCCTGGGCGAGCGAGTGACCTTCACATGCAAAGCATCCCAGG ACATCAACACATACCTGTCTTGGTTTCAGCAGAAGCCAGGCAA AAGCCCCAAGACCCTGATCTACCGGGCCAATAGACTGGTGGA CGGGGTCCCCAGCAGATTCTCCGGATCTGGCAGTGGGCAGGA TTACTCCCTGACCATCAGCTCCCTGGAGTATGAAGACATGGGC ATCTACTATTGCCTGCAGTATGATGAGTTCCCTCTGACCTTTG GAGCAGGCACAAAACTGGAACTGAAG (SEQ ID NO: 16) Amino acid sequence of the light chain variable region of 14C12: (107 aa) DIKMTQSPSSMYASLGERVTFTCKASQDINTYLSWFQQKPG KSPKTLIYRANRLVDGVPSRFSGSGSGQDYSLTISSLEYEDMGIYY CLQYDEFPLTFGAGTKLELK (SEQ ID NO: 17) (2) Heavy and light chain variable region sequences and heavy and light chain sequences of humanized monoclonal antibody 14C12H1L1 Nucleotide sequence of the heavy chain variable region 14C12H1v of 14C12H1L1: (354 bp) GAAGTGCAGCTGGTCGAGTCTGGGGGAGGGCTGGTGCAG CCCGGCGGGTCACTGCGACTGAGCTGCGCAGCTTCCGGATTC GCCTTTAGCTCCTACGACATGTCCTGGGTGCGACAGGCACCA GGAAAGGGACTGGATTGGGTCGCTACTATCTCAGGAGGCGGG AGATACACCTACTATCCTGACAGCGTCAAGGGCCGGTTCACA ATCTCTAGAGATAACAGTAAGAACAATCTGTATCTGCAGATGA ACAGCCTGAGGGCTGAGGACACCGCACTGTACTATTGTGCCA ACCGCTACGGGGAAGCATGGTTTGCCTATTGGGGGCAGGGAA CCCTGGTGACAGTCTCTAGT (SEQ ID NO: 18) Amino acid sequence of the heavy chain variable region 14C12H1v of 14C12H1L1: (118 aa) EVQLVESGGGLVQPGGSLRLSCAASGFAFSSYDMSWVRQAP GKGLDWVATISGGGRYTYYPDSVKGRFTISRDNSKNNLYLQMNS LRAEDTALYYCANRYGEAWFAYWGQGTLVTVSS (SEQ ID NO: 19) Nucleotide sequence of the light chain variable region 14C12L1v of 14C12H1L1: (321 bp) GACATTCAGATGACTCAGAGCCCCTCCTCCATGTCCGCCT CTGTGGGCGACAGGGTCACCTTCACATGCCGCGCTAGTCAGG ATATCAACACCTACCTGAGCTGGTTTCAGCAGAAGCCAGGGA AAAGCCCCAAGACACTGATCTACCGGGCTAATAGACTGGTGT CTGGAGTCCCAAGTCGGTTCAGTGGCTCAGGGAGCGGACAGG ACTACACTCTGACCATCAGCTCCCTGCAGCCTGAGGACATGG CAACCTACTATTGCCTGCAGTATGATGAGTTCCCACTGACCTT TGGCGCCGGGACAAAACTGGAGCTGAAG (SEQ ID NO: 20) Amino acid sequence of the light chain variable region 14C12L1v of 14C12H1L1: (107 aa) DIQMTQSPSSMSASVGDRVTFTCRASQDINTYLSWFQQKPGK SPKTLIYRANRLVSGVPSRFSGSGSGQDYTLTISSLQPEDMATYYC LQYDEFPLTFGAGTKLELK (SEQ ID NO: 21) Nucleotide sequence of the heavy chain 14C12H1 of 14C12H1L1: (1344 bp) GAAGTGCAGCTGGTCGAGTCTGGGGGAGGGCTGGTGCAG CCCGGCGGGTCACTGCGACTGAGCTGCGCAGCTTCCGGATTC GCCTTTAGCTCCTACGACATGTCCTGGGTGCGACAGGCACCA GGAAAGGGACTGGATTGGGTCGCTACTATCTCAGGAGGCGGG AGATACACCTACTATCCTGACAGCGTCAAGGGCCGGTTCACA ATCTCTAGAGATAACAGTAAGAACAATCTGTATCTGCAGATGA ACAGCCTGAGGGCTGAGGACACCGCACTGTACTATTGTGCCA ACCGCTACGGGGAAGCATGGTTTGCCTATTGGGGGCAGGGAA CCCTGGTGACAGTCTCTAGTGCCAGCACCAAAGGACCTAGCG TGTTTCCTCTCGCCCCCTCCTCCAAAAGCACCAGCGGAGGAAC CGCTGCTCTCGGATGTCTGGTGAAGGACTACTTCCCTGAACCC GTCACCGTGAGCTGGAATAGCGGCGCTCTGACAAGCGGAGTC CATACATTCCCTGCTGTGCTGCAAAGCAGCGGACTCTATTCCC TGTCCAGCGTCGTCACAGTGCCCAGCAGCAGCCTGGGCACCC AGACCTACATCTGTAACGTCAACCACAAGCCCTCCAACACCAA GGTGGACAAGAAAGTGGAGCCCAAATCCTGCGACAAGACACA CACCTGTCCCCCCTGTCCTGCTCCCGAACTCCTCGGAGGCCCT AGCGTCTTCCTCTTTCCTCCCAAACCCAAGGACACCCTCATGA TCAGCAGAACCCCTGAAGTCACCTGTGTCGTCGTGGATGTCA GCCATGAGGACCCCGAGGTGAAATTCAACTGGTATGTCGATG GCGTCGAGGTGCACAACGCCAAAACCAAGCCCAGGGAGGAAC AGTACAACTCCACCTACAGGGTGGTGTCCGTGCTGACAGTCC TCCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGG TGTCCAACAAGGCTCTCCCTGCCCCCATTGAGAAGACCATCAG CAAGGCCAAAGGCCAACCCAGGGAGCCCCAGGTCTATACACT GCCTCCCTCCAGGGACGAACTCACCAAGAACCAGGTGTCCCT GACCTGCCTGGTCAAGGGCTTTTATCCCAGCGACATCGCCGT CGAGTGGGAGTCCAACGGACAGCCCGAGAATAACTACAAGAC CACCCCTCCTGTCCTCGACTCCGACGGCTCCTTCTTCCTGTAC AGCAAGCTGACCGTGGACAAAAGCAGGTGGCAGCAGGGAAAC GTGTTCTCCTGCAGCGTGATGCACGAAGCCCTCCACAACCACT ACACCCAGAAAAGCCTGTCCCTGAGCCCCGGCAAA (SEQ ID NO: 22) Amino acid sequence of the heavy chain 14C12H1 of 14C12H1L1: (448 aa) EVQLVESGGGLVQPGGSLRLSCAASGFAFSSYDMSWVRQAP GKGLDWVATISGGGRYTYYPDSVKGRFTISRDNSKNNLYLQMNS LRAEDTALYYCANRYGEAWFAYWGQGTLVTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKS CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVV DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPP VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS LSLSPGK (SEQ ID NO: 23) Nucleotide sequence of the light chain 14C12L1 of 14C12H1L1: (642 bp) GACATTCAGATGACTCAGAGCCCCTCCTCCATGTCCGCCT CTGTGGGCGACAGGGTCACCTTCACATGCCGCGCTAGTCAGG ATATCAACACCTACCTGAGCTGGTTTCAGCAGAAGCCAGGGA AAAGCCCCAAGACACTGATCTACCGGGCTAATAGACTGGTGT CTGGAGTCCCAAGTCGGTTCAGTGGCTCAGGGAGCGGACAGG ACTACACTCTGACCATCAGCTCCCTGCAGCCTGAGGACATGG CAACCTACTATTGCCTGCAGTATGATGAGTTCCCACTGACCTT TGGCGCCGGGACAAAACTGGAGCTGAAGCGAACTGTGGCCGC TCCCTCCGTCTTCATTTTTCCCCCTTCTGACGAACAGCTGAAA TCAGGCACAGCCAGCGTGGTCTGTCTGCTGAACAATTTCTACC CTAGAGAGGCAAAAGTGCAGTGGAAGGTCGATAACGCCCTGC AGTCCGGCAACAGCCAGGAGAGTGTGACTGAACAGGACTCAA AAGATAGCACCTATTCCCTGTCTAGTACACTGACTCTGTCCAA GGCTGATTACGAGAAGCACAAAGTGTATGCATGCGAAGTGAC ACATCAGGGACTGTCAAGCCCCGTGACTAAGTCTTTTAACCGG GGCGAATGT (SEQ ID NO: 24) Amino acid sequence of the light chain 14C12L1 of 14C12H1L1: (214 aa) DIQMTQSPSSMSASVGDRVTFTCRASQDINTYLSWFQQKPGK SPKTLIYRANRLVSGVPSRFSGSGSGQDYTLTISSLQPEDMATYYC LQYDEFPLTFGAGTKLELKRTVAAPSVFIFPPSDEQLKSGTASVV CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 25) The CDRs of the antibodies 14C12 and 14C12H1L1 are identical as follows (according to the IMGT numbering system): HCDR1: GFAFSSYD (SEQ ID NO: 26) HCDR2: ISGGGRYT (SEQ ID NO: 27) HCDR3: ANRYGEAWFAY (SEQ ID NO: 28) LCDR1: QDINTY (SEQ ID NO: 29) LCDR2: RAN (SEQ ID NO: 30) LCDR3: LQYDEFPLT (SEQ ID NO: 31) Heavy and light chain variable region sequences of 14C12H1L1(M) 14C12H1L1(M) was obtained by mutating certain amino acids in the framework region (light chain) on the basis of 14C12H1L1. The heavy chain variable region 14C12H1(M) of 14C12H1L1(M) is identical to the heavy chain variable region 14C12H1 of 14C12H1L1; that is, both heavy chain variable regions have the amino acid sequence set forth in SEQ ID NO: 19. Light chain variable region 14C12L1(M) of 14C12H1L1(M): DIQMTQSPSSMSASVGDRVTFTCRASQDINTYLSWFQQKPGK SPKTLIYRANRLVSGVPSRFSGSGSGQDYTLTISSLQPEDMATYYC LQYDEFPLTFGAGTKLELKR (SEQ ID NO: 38) Preparation Example 3:  Design of Humanized Antibody 14C12H1L1(hG4DM) On the basis of 14C12H1L1, the inventors obtained a humanized antibody 14C12H1L1(hG4DM) with constant region mutations by using Ig gamma-4 chain C region as a heavy chain constant region and introducing a phenylalanine-to-alanine point mutation at position 234 (F234A) and a leucine-to-alanine point mutation at position 235 (L235A) in the heavy chain constant region while keeping the antibody variable region unchanged. The amino acid sequence of the heavy chain 14C12H1(hG4DM) of 14C12H1L1(hG4DM) is set forth in SEQ ID NO: 32, and the amino acid sequence of the light chain thereof is set forth in SEQ ID NO: 25. Amino acid sequence of the heavy chain of 14C12H1L1(hG4DM) EVQLVESGGGLVQPGGSLRLSCAASGFAFSSYDMSWVRQAP GKGLDWVATISGGGRYTYYPDSVKGRFTISRDNSKNNLYLQMNS LRAEDTALYYCANRYGEAWFAYWGQGTLVTVSSASTKGPSVFPL APCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESK YGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS QEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQ EEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD SDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLGK (SEQ ID NO: 32) The amino acid sequence of the light chain of 14C12H1L1(hG4DM) is identical to the amino acid sequence of the light chain 14C12L1 of 14C12H1L1, as set forth in SEQ ID NO: 25. Preparation Example 4: Sequence Design of Humanized Antibody 14C12H1L1(hG1TM) On the basis of the humanized antibody 14C12H1L1, the inventors obtained a humanized mutant 14C12H1L1(hG1TM) by introducing a leucine-to-alanine point mutation at position 234 (L234A), a leucine-to-alanine point mutation at position 235 (L235A), and a glycine-to-alanine point mutation at position 237 (G237A) in the hinge region of the heavy chain according to the EU numbering system. Nucleotide sequence of the heavy chain 14C12H1(hG1TM) of 14C12H1L1(hG1TM): (1344 bp) GAAGTGCAGCTGGTCGAGTCTGGGGGAGGGCTGGTGCAG CCCGGCGGGTCACTGCGACTGAGCTGCGCAGCTTCCGGATTC GCCTTTAGCTCCTACGACATGTCCTGGGTGCGACAGGCACCA GGAAAGGGACTGGATTGGGTCGCTACTATCTCAGGAGGCGGG AGATACACCTACTATCCTGACAGCGTCAAGGGCCGGTTCACA ATCTCTAGAGATAACAGTAAGAACAATCTGTATCTGCAGATGA ACAGCCTGAGGGCTGAGGACACCGCACTGTACTATTGTGCCA ACCGCTACGGGGAAGCATGGTTTGCCTATTGGGGGCAGGGAA CCCTGGTGACAGTCTCTAGTGCCAGCACCAAAGGGCCCAGCG TGTTTCCTCTCGCCCCCTCCTCCAAAAGCACCAGCGGAGGAAC CGCTGCTCTCGGATGTCTGGTGAAGGACTACTTCCCTGAACCC GTCACCGTGAGCTGGAATAGCGGCGCTCTGACAAGCGGAGTC CATACATTCCCTGCTGTGCTGCAAAGCAGCGGACTCTATTCCC TGTCCAGCGTCGTCACAGTGCCCAGCAGCAGCCTGGGCACCC AGACCTACATCTGTAACGTCAACCACAAGCCCTCCAACACCAA GGTGGACAAGAAAGTGGAGCCCAAATCCTGCGACAAGACACA CACCTGTCCCCCCTGTCCTGCTCCCGAAGCTGCTGGAGCCCCT AGCGTCTTCCTCTTTCCTCCCAAACCCAAGGACACCCTCATGA TCAGCAGAACCCCTGAAGTCACCTGTGTCGTCGTGGATGTCA GCCATGAGGACCCCGAGGTGAAATTCAACTGGTATGTCGATG GCGTCGAGGTGCACAACGCCAAAACCAAGCCCAGGGAGGAAC AGTACAACTCCACCTACAGGGTGGTGTCCGTGCTGACAGTCC TCCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGG TGTCCAACAAGGCTCTCCCTGCCCCCATTGAGAAGACCATCAG CAAGGCCAAAGGCCAACCCAGGGAGCCCCAGGTCTATACACT GCCTCCCTCCAGGGACGAACTCACCAAGAACCAGGTGTCCCT GACCTGCCTGGTCAAGGGCTTTTATCCCAGCGACATCGCCGT CGAGTGGGAGTCCAACGGACAGCCCGAGAATAACTACAAGAC CACCCCTCCTGTCCTCGACTCCGACGGCTCCTTCTTCCTGTAC AGCAAGCTGACCGTGGACAAAAGCAGGTGGCAGCAGGGAAAC GTGTTCTCCTGCAGCGTGATGCACGAAGCCCTCCACAACCACT ACACCCAGAAAAGCCTGTCCCTGAGCCCCGGCAAA (SEQ ID NO: 33) Amino acid sequence of the heavy chain 14C12H1(hG1TM) of 14C12H1L1(hG1TM): (448 aa) EVQLVESGGGLVQPGGSLRLSCAASGFAFSSYDMSWVRQAP GKGLDWVATISGGGRYTYYPDSVKGRFTISRDNSKNNLYLQMNS LRAEDTALYYCANRYGEAWFAYWGQGTLVTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKS CDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVV DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPP VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS LSLSPGK (SEQ ID NO: 34) The nucleotide sequence of the light chain of 14C12H1L1(hG1TM) is set forth in SEQ ID NO: 24. The amino acid sequence of the light chain of 14C12H1L1(hG1TM) is identical to the amino acid sequence of the light chain 14C12L1 of 14C12H1L1, as set forth in SEQ ID NO: 25. Preparation Example 5: Design and Preparation of Anti-LAG3 / PD-1 Bispecific Antibodies 1. Sequence design The structures of the bispecific antibodies BS-PL021A, Bs-PL022B, BS-PL023C, Bs-PLV02, and Bi-PGV02 of the present disclosure are in the Morrison format (IgG-scFv); that is, the C-termini of two heavy chains of an IgG antibody are each linked to an scFv fragment of another antibody. The main composition design of the heavy and light chains is as shown in Table 1 below. Table 1: Composition design of heavy and light chains of bispecific antibodies Bispecific antibody No. Immunoglobulin portion Linker scFv portion Heavy chain Light chain Heavy chain variable region Linker Light chain variable region BS-PL021A H7(hGlTM) (SEQ ID NO: 11) L8 (SEQ ID NO: 12) (GGGGS)3 14C12H1V (SEQ ID NO: 19) (GGGGS)4 14C12Ll(M)v (SEQ ID NO: 38) BS-PL022B H7(hGlTM) (SEQ ID NO: 11) L8 (SEQ ID NO: 12) (GGGGS)4 14C12H1V (SEQ ID NO: 19) (GGGGS)4 14C12Ll(M)v (SEQ ID NO: 38) BS-PL023C 14C12Hl(hGlTM) (SEQ ID NO: 34) 14C12L1 (SEQ ID NO: 25) (GGGGS)4 H7v (SEQ ID NO: 2) (GGGGS)4 L8v (SEQ ID NO: 4) Bs-PLV02 14C12Hl(hG4DM) (SEQ ID NO: 32) 14C12L1 (SEQ ID NO: 25) (GGGGS)4G H7v (SEQ ID NO: 2) (GGGGS)4 L8v (SEQ ID NO: 4) Bi-PGV02 H7(hG4DM) (SEQ ID NO: 13) L8 (SEQ ID (GGGGS)4 14C12H1V (SEQ ID NO: (GGGGS)4 14C12L1V (SEQ ID NO: NO: 12) 19) 21) In Table 1 above: (1) Amino acid sequence of linker (GGGGS)3: GGGGSGGGGSGGGGS (SEQ ID NO: 35) Amino acid sequence of linker (GGGGS)4: GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 36) Amino acid sequence of linker (GGGGS)4G: GGGGSGGGGSGGGGSGGGGSG (SEQ ID NO: 37) (2) Those with a “v” label at the lower right corner refer to the variable region of the corresponding heavy chain or the variable region of the corresponding light chain. For those without a “v” label, the corresponding heavy or light chain is the full length comprising the constant region. The amino acid sequences of these variable regions or full-length sequences and the nucleotide sequences encoding them were as described in the corresponding sequences in the preparation examples described above. 2. Expression and purification of antibodies The heavy chain cDNA sequences and the light chain cDNA sequences of Bs-PL021A, Bs-PL022B, Bs-PL023C, Bs-PLV02, and Bi-PGV02 were separately cloned into pUC57simple vectors (supplied by GenScript), and plasmids pUC57simple-Bs-PL021AH / pUC57simple-Bs-PL021AL,     pUC57simple-Bs-PL022BH / pUC57simple-Bs-PL022BL, pUC57simple-Bs-PL023CH / pUC57simple-Bs-PL023CL, pUC57simple- Bs-PLV02H / pUC57simple-Bs-PLV02L, and pUC57simple-Bi-PGV02H / pUC57simple-Bi-PGV02L  were obtained, respectively. The plasmids   pUC57simple-Bs-PL021AH / pUC57simple-Bs- PL021AL,     pUC57simple-Bs-PL022BH / pUC57simple-Bs-PL022BL, pUC57simple-Bs-PL023CH / pUC57simple-Bs-PL023CL, pUC57simple-Bs-PLV02H / pUC57simple-Bs-PLV02L and pUC57simple-Bi-PGV02H / pUC57simple-Bi-PGV02L were each digested (HindIII&EcoRI). The heavy and light chains isolated by electrophoresis were separately subcloned into pcDNA3.1 vectors, and recombinant plasmids were extracted to co-transfect 293F cells. After 7 days of cell culture, the culture solution was separated by high-speed centrifugation, and the supernatant was concentrated and loaded onto a HiTrap MabSelect SuRe column. The protein was eluted in one step with an elution buffer. The target sample was isolated and buffer-exchanged into PBS. Preparation Example 6: Preparation of Anti-Human CD47 Antibody 6F7 1. Preparation of hybridoma cell line 6F7 The antigen used for preparing the anti-CD47 antibody with the hybridoma cell line 6F7 was CD47 IgV TEV-His (including human CD47 mature peptide of positions 19-141 of GenbankID: NP 942088.1 and TEV (amino acid sequence: ENLYFQG, SEQ ID NO: 72)-his tag fusion protein, synthesized by Akeso Biopharma Inc.) and 3T3-CD47 cells (NIH / 3T3, manufacturer: ATCC, Cat. No: CRL-1658; the human CD47 mature peptide was transfected into the cells on the basis of NIH / 3T3 to construct a cell line stably expressing 3T3-CD47). Spleen cells of immunized mice were fused with myeloma cells of mice to prepare hybridoma cells. With CD47 IgV TEV-His and 3T3-CD47 cells separately taken as antigens, the hybridoma cells were screened by indirect ELISA to obtain hybridoma cells capable of secreting antibodies capable of specifically binding to CD47. The hybridoma cells obtained by ELISA screening were screened by competitive ELISA to obtain a hybridoma cell line capable of secreting a monoclonal antibody capable of competing for binding to human CD47 IgV TEV-His with the receptor human  SIRPaECD-hFc-Biotin (SIRPaECD refers to the extracellular region of SIRPa, positions 31 -373 of protein GenBank Accession No. NP_542970.1; hFc refers to a human IgG Fc purification tag, specifically to Ig gamma-1 chain C region, positions 114-330 of GenbankID: P01857), which was then subjected to a limiting dilution assay to obtain a stable hybridoma cell line. The hybridoma cell line above was named hybridoma cell line LT012, and the monoclonal antibody secreted by the cell line was named 6F7. Hybridoma cell line LT012 was deposited at China Center for Type Culture Collection (CCTCC) on June 21, 2018 with a CCTCC designation of CCTCC NO. C2018135, the depository address being Wuhan University, Wuhan, China, postal code: 430072. 2. Preparation of anti-CD47 antibody 6F7 The cell line LT012 prepared above was cultured with a chemical defined medium (CD medium; containing 1% penicillin-streptomycin) in a 5% CO2, 37 °C incubator. After 7 days, the supernatant was collected and purified by high-speed centrifugation and vacuum filtration through a microfiltration membrane and through a HiTrap protein A HP column to obtain antibody 6F7. Preparation Example 7: Sequence Analysis of Anti-CD47 Antibody 6F7 mRNA was extracted from the cell line LT012 cultured in Preparation Example 6 according to the method described in the manual of RNAprep pure Cell / Bacteria Kit (Tiangen, Cat. No.: DP430). cDNA was synthesized according to the instructions of an Invitrogen SuperScript® III First-Strand Synthesis System for RT-PCR kit and amplified by PCR. The PCR-amplified products were directly subjected to TA cloning according to the manual of the pEASY-T1 Cloning Kit (Transgen CT101). The TA cloning products were directly sequenced, and the sequencing results are as follows: The nucleotide sequence of the heavy chain variable region is set forth in SEQ ID NO: 49 with a length of 351 bp. The encoded amino acid sequence is set forth in SEQ ID NO: 50 with a length of 117 aa, and the sequences of heavy chain CDR1, CDR2, and CDR3 are set forth in SEQ ID NOs: 53, 54, and 55, respectively. The nucleotide sequence of the light chain variable region is set forth in SEQ ID NO: 51 with a length of 321 bp. The encoded amino acid sequence is set forth in SEQ ID NO: 52 with a length of 107 aa, and the sequences of light chain CDR1, CDR2, and CDR3 are set forth in SEQ ID NOs: 56, 57, and 58, respectively. 6F7 heavy chain variable region: CAGGTGCAGCTGCAGCAGCCAGGAGCAGAGCTGGTGAGG CCAGGAGCATCCGTGAAGCTGTCTTGTAAGGCCAGCGGCTAC ACCTTCACATCCTATTGGATGAACTGGGTGAAGCAGAGGCCT GGACAGGGACTGGAGTGGATCGGCATGATCGACCCAAGCGAT TCCGAGACCCACAACAATCAGATGTTTAAGGACAAGGCCACC CTGACAGTGGATAAGAGCTCCAATACCGCCTACATGCACCTGT CTAGCCTGACATCTGAGGACAGCGCCGTGTATCACTGCGCCC GGCTGTACAGATGGTATTTTGACGTGTGGGGAGCAGGAACCA CAGTGACCGTGTCCTCT (SEQ ID NO: 49) QVQLQQPGAELVRPGASVKLSCKASGYTFTSYWMNWVKQR PGQGLEWIGMIDPSDSETHNNQMFKDKATLTVDKSSNTAYMHLS SLTSEDSAVYHCARLYRWYFDVWGAGTTVTVSS (SEQ ID NO: 50) 6F7 light chain variable region: AACATCGTGATGACCCAGTCCCCCAAGTCTATGAGCATGT CCCTGGGCGAGAGGGTGACCCTGTCCTGTAAGGCCTCTGAGA TCGTGGGCACATACGTGTCTTGGTTTCAGCAGAAGCCACACC AGAGCCCCAAGCTGCTGATCTACGGCGCCTCCAATCGGTATA CAGGCGTGCCTGACAGATTCACCGGCTCTGGCAGCGCCACAG ACTTCACCCTGACAATCTCTAACGTGCAGGCCGAGGACCTGG CCGATTATCACTGCGGCCAGAGCTACAATTTCCCTTATACCTT TGGCGGCGGCACAAAGCTGGAGATCAAG (SEQ ID NO: 51) NIVMTQSPKSMSMSLGERVTLSCKASEIVGTYVSWFQQKPH QSPKLLIYGASNRYTGVPDRFTGSGSATDFTLTISNVQAEDLADY HCGQSYNFPYTFGGGTKLEIK (SEQ ID NO: 52) The 6 CDRs of antibody 6F7 are defined by the IMGT numbering system as follows: HCDR1: GYTFTSYW (SEQ ID NO: 53) HCDR2: IDPSDSET (SEQ ID NO: 54) HCDR3: ARLYRWYFDV (SEQ ID NO: 55) LCDR1: EIVGTY (SEQ ID NO: 56) LCDR2: GAS (SEQ ID NO: 57) LCDR3: GQSYNFPYT (SEQ ID NO: 58) Preparation Example 8: Light and Heavy Chain Design and Preparation of Humanized Anti-Human CD47 Antibodies 6F7 H1L1(hG4), 6F7 H2L2(hG4), and 6F7 H3L3(hG4) 1. Light and heavy chain design of humanized anti-human CD47 antibodies 6F7 H1L1(hG4), 6F7 H2L2(hG4), and 6F7 H3L3(hG4) Based on the three-dimensional crystal structure of human CD47 protein (Hage T, Reinemer P, Sebald W. Crystals of a 1:1 complex between human interleukin-4 and the extracellular domain of its receptor alpha chain. Eur J Biochem. 1998; 258(2):831-6.) and the sequence of antibody 6F7 obtained in Preparation Example 7, the variable region sequences of antibodies 6F7 H1L1, 6F7 H2L2, and 6F7 H3L3 were obtained by computer modeling and mutation design (antibody constant region sequences from NCBI database: the heavy chain constant region is Ig gamma-4 chain C region, ACCESSION: P01861.1; the light chain constant region is Ig kappa chain C region, ACCESSION: P01834). The designed variable region sequences are as follows: (1) Heavy and light chain variable region sequences of humanized monoclonal antibody 6F7 H1L1 The nucleotide sequence of the heavy chain variable region is set forth in SEQ ID NO: 59 with a length of 351 bp. The encoded amino acid sequence is set forth in SEQ ID NO: 60 with a length of 117 aa, and the sequences of heavy chain CDR1, CDR2, and CDR3 are set forth in SEQ ID NOs: 53, 54, and 55, respectively. The nucleotide sequence of the light chain variable region is set forth in SEQ ID NO: 61 with a length of 321 bp. The encoded amino acid sequence is set forth in SEQ ID NO: 62 with a length of 107 aa, and the sequences of light chain CDR1, CDR2, and CDR3 are set forth in SEQ ID NOs: 56, 57, and 58, respectively. (2) Heavy and light chain variable region sequences of humanized monoclonal antibody 6F7 H2L2 The nucleotide sequence of the heavy chain variable region is set forth in SEQ ID NO: 63 with a length of 351 bp. The encoded amino acid sequence is set forth in SEQ ID NO: 64 with a length of 117 aa, and the sequences of heavy chain CDR1, CDR2, and CDR3 are set forth in SEQ ID NOs: 53, 54, and 55, respectively. The nucleotide sequence of the light chain variable region is set forth in SEQ ID NO: 65 with a length of 321 bp. The encoded amino acid sequence is set forth in SEQ ID NO: 66 with a length of 107 aa, and the sequences of light chain CDR1, CDR2, and CDR3 are set forth in SEQ ID NOs: 56, 57, and 58, respectively. (3) Heavy and light chain variable region sequences of humanized monoclonal antibody 6F7 H3L3 The nucleotide sequence of the heavy chain variable region is set forth in SEQ ID NO: 67 with a length of 351 bp. The encoded amino acid sequence is set forth in SEQ ID NO: 68 with a length of 117 aa, and the sequences of heavy chain CDR1, CDR2, and CDR3 are set forth in SEQ ID NOs: 53, 54, and 55, respectively. The nucleotide sequence of the light chain variable region is set forth in SEQ ID NO: 69 with a length of 321 bp. The encoded amino acid sequence is set forth in SEQ ID NO: 70 with a length of 107 aa, and the sequences of light chain CDR1, CDR2, and CDR3 are set forth in SEQ ID NOs: 56, 57, and 58, respectively. 6F7H1: CAGGTGCAGCTGGTGCAGAGCGGAGCAGAGGTGGTGAAG CCAGGAGCCTCTGTGAAGCTGAGCTGTAAGGCCTCCGGCTAC ACCTTCACAAGCTATTGGATGAACTGGGTGCGGCAGAGACCA GGACAGGGACTGGAGTGGATCGGAATGATCGACCCTTCCGAT TCTGAGACCCACAATGCCCAGAAGTTTCAGGGCAAGGCCACC CTGACAGTGGACAAGAGCACCTCCACAGCCTACATGCACCTG AGCTCCCTGCGGTCCGAGGACACAGCCGTGTACTATTGCGCC AGGCTGTACCGCTGGTATTTTGACGTGTGGGGAGCAGGAACC ACAGTGACCGTGTCTAGC (SEQ ID NO: 59) QVQLVQSGAEVVKPGASVKLSCKASGYTFTSYWMNWVRQR PGQGLEWIGMIDPSDSETHNAQKFQGKATLTVDKSTSTAYMHLS SLRSEDTAVYYCARLYRWYFDVWGAGTTVTVSS (SEQ ID NO: 60) 6F7L1: AACATCGTGATGACCCAGTCCCCAGCCACAATGTCTATGA GCCCAGGAGAGAGGGTGACCCTGTCCTGTAGAGCCTCTGAGA TCGTGGGCACATACGTGTCTTGGTTTCAGCAGAAGCCAGGAC AGGCACCTAGGCTGCTGATCTACGGAGCAAGCAACAGGTATA CCGGAGTGCCAGCACGCTTCTCCGGCTCTGGCAGCGGCACAG ACTTTACCCTGACAATCAGCTCCGTGCAGCCTGAGGACCTGG CCGATTATCACTGCGGCCAGTCTTACAATTTCCCATATACCTT TGGCGGCGGCACAAAGCTGGAGATCAAG (SEQ ID NO: 61) NIVMTQSPATMSMSPGERVTLSCRASEIVGTYVSWFQQKPG QAPRLLIYGASNRYTGVPARFSGSGSGTDFTLTISSVQPEDLADYH CGQSYNFPYTFGGGTKLEIK (SEQ ID NO: 62) 6F7H2: CAGGTGCAGCTGGTGCAGAGCGGAGCAGAGGTGGTGAAG CCAGGAGCCTCTGTGAAGGTGAGCTGTAAGGCCTCCGGCTAC ACCTTCACATCCTATTGGATGAACTGGGTGCGGCAGAGACCA GGACAGGGACTGGAGTGGATCGGAATCATCGACCCTTCCGAT TCTGAGACCTCTAATGCCCAGAAGTTTCAGGGCCGGGTGACC CTGACAGTGGACAAGAGCACCTCCACAGCCTACATGCACCTG AGCTCCCTGAGGAGCGAGGACACAGCCGTGTACTATTGCGCC AGGCTGTACCGCTGGTATTTTGACGTGTGGGGAGCAGGAACC ACAGTGACCGTGTCTAGC (SEQ ID NO: 63) QVQLVQSGAEVVKPGASVKVSCKASGYTFTSYWMNWVRQR PGQGLEWIGIIDPSDSETSNAQKFQGRVTLTVDKSTSTAYMHLSS LRSEDTAVYYCARLYRWYFDVWGAGTTVTVSS (SEQ ID NO: 64) 6F7L2: AACATCGTGATGACCCAGTCCCCAGCCACACTGTCTCTGA GCCCAGGAGAGAGGGTGACCCTGTCCTGTAGAGCCTCTGAGA TCGTGGGCACATACGTGTCTTGGTTTCAGCAGAAGCCAGGAC AGGCACCTAGGCTGCTGATCTATGGCGCCAGCAACAGGGCAA CCGGCATCCCCGCACGCTTCTCCGGCTCTGGCAGCGGCACAG ACTTTACCCTGACAATCAGCTCCCTGCAGCCTGAGGACCTGGC CGATTACTATTGCGGCCAGTCTTACAATTTCCCATATACCTTT GGCGGCGGCACAAAGCTGGAGATCAAG (SEQ ID NO: 65) NIVMTQSPATLSLSPGERVTLSCRASEIVGTYVSWFQQKPGQ APRLLIYGASNRATGIPARFSGSGSGTDFTLTISSLQPEDLADYYC GQSYNFPYTFGGGTKLEIK (SEQ ID NO: 66) 6F7H3: CAGGTGCAGCTGGTGCAGAGCGGAGCAGAGGTGGTGAAG CCAGGAGCCTCTGTGAAGGTGAGCTGTAAGGCCTCCGGCTAC ACCTTCACATCCTATTGGATGAACTGGGTGCGGCAGGCACCA GGACAGGGACTGGAGTGGATCGGCATCATCGACCCTTCCGAT TCTGAGACCTCTTACGCCCAGAAGTTTCAGGGCAGGGTGACC CTGACAGTGGACAAGAGCACCTCCACAGCCTATATGGAGCTG AGCTCCCTGCGCAGCGAGGACACAGCCGTGTACTATTGCGCC CGGCTGTACAGATGGTATTTTGACGTGTGGGGAGCAGGAACC ACAGTGACCGTGTCTAGC (SEQ ID NO: 67) QVQLVQSGAEVVKPGASVKVSCKASGYTFTSYWMNWVRQA PGQGLEWIGIIDPSDSETSYAQKFQGRVTLTVDKSTSTAYMELSSL RSEDTAVYYCARLYRWYFDVWGAGTTVTVSS (SEQ ID NO: 68) 6F7L3: AACATCGTGATGACCCAGTCCCCAGCCACACTGTCTCTGA GCCCAGGAGAGAGGGTGACCCTGTCCTGTAGAGCCTCTGAGA TCGTGGGCACATACCTGTCTTGGTATCAGCAGAAGCCAGGAC AGGCACCTAGGCTGCTGATCTACGGAGCCAGCACCAGGGCAA CAGGCATCCCCGCACGCTTCTCCGGCTCTGGCAGCGGCACCG ACTTTACCCTGACAATCAGCTCCCTGCAGCCTGAGGATTTTGC CGTGTACTATTGCGGCCAGTCTTACAATTTCCCATATACCTTT GGCGGCGGCACAAAGCTGGAGATCAAG (SEQ ID NO: 69) NIVMTQSPATLSLSPGERVTLSCRASEIVGTYLSWYQQKPGQ APRLLIYGASTRATGIPARFSGSGSGTDFTLTISSLQPEDFAVYYC GQSYNFPYTFGGGTKLEIK (SEQ ID NO: 70) 2. Preparation of humanized antibodies 6F7 H1L1(hG4), 6F7 H2L2(hG4), and 6F7 H3L3(hG4) The heavy chain constant regions are all Ig gamma-4 chain C region, SEQ ID NO: 71; the light chain constant regions are all Ig kappa chain C region, SEQ ID NO: 40. The heavy chain cDNA and light chain cDNA of 6F7 H1L1(hG4), the heavy chain cDNA and light chain cDNA of 6F7 H2L2(hG4), and the heavy chain cDNA and light chain cDNA of 6F7 H3L3(hG4) were separately cloned into pUC57simple vectors (provided by GenScript) to obtain pUC57simple-6F7H1 and pUC57simple-6F7L1, pUC57simple-6F7H2 and pUC57simple-6F7L2, and pUC57simple-6F7H3 and pUC57simple-6F7L3. With reference to the standard techniques described in Molecular Cloning: A Laboratory Manual (Third Edition), the heavy and light chain full-length genes synthesized by EcoRI&HindIII digestion of genes were subcloned into an expression vector pcDNA3.1 through digestion by a restriction enzyme (EcoRI&HindIII) to obtain expression plasmids pcDNA3.1-6F7H1, pcDNA3.1-6F7L1, pcDNA3.1-6F7H2, pcDNA3.1-6F7L2, pcDNA3.1- 6F7H3, and pcDNA3.1-6F7L3, and the heavy and light chain genes of the recombinant expression plasmids were further sequenced. Subsequently, the designed recombinant plasmid gene combinations containing the corresponding light and heavy chains (pcDNA3.1-6F7H1 / pcDNA3.1-6F7L1, pcDNA3.1-6F7H2 / pcDNA3.1-6F7L2, and pcDNA3.1-6F7H3 / pcDNA3.1-6F7L3) were separately co-transfected into 293F cells, and then the culture solutions were collected and purified. After the sequences were verified, endotoxin-free expression plasmids were prepared and transiently transfected into HEK293 cells for antibody expression. The cell cultures were collected after 7 days of culture, and subjected to affinity purification on a Protein A column (MabSelect SURE (GE)) to obtain humanized antibodies. Heavy chain constant region Ig gamma-4 chain C region ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNS GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHK PSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMIS RTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQ PREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEA LHNHYTQKSLSLSLGK (SEQ ID NO: 71) Light chain constant region Ig kappa chain C region RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWK VDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 40) Amino acid sequence of 6F7 H1L1(hG4) heavy chain QVQLVQSGAEVVKPGASVKLSCKASGYTFTSYWMNWVRQR PGQGLEWIGMIDPSDSETHNAQKFQGKATLTVDKSTSTAYMHLS SLRSEDTAVYYCARLYRWYFDVWGAGTTVTVSSASTKGPSVFPL APCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESK YGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS QEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQ EEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD SDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLGK (SEQ ID NO: 74) Amino acid sequence of 6F7 H1L1(hG4) light chain NIVMTQSPATMSMSPGERVTLSCRASEIVGTYVSWFQQKPG QAPRLLIYGASNRYTGVPARFSGSGSGTDFTLTISSVQPEDLADYH CGQSYNFPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVV CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 75) Amino acid sequence of 6F7 H2L2(hG4) heavy chain QVQLVQSGAEVVKPGASVKVSCKASGYTFTSYWMNWVRQR PGQGLEWIGIIDPSDSETSNAQKFQGRVTLTVDKSTSTAYMHLSS LRSEDTAVYYCARLYRWYFDVWGAGTTVTVSSASTKGPSVFPLA PCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL QSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKY GPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQ EDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQE EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS DGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLS LGK (SEQ ID NO: 76) Amino acid sequence of 6F7 H2L2(hG4) light chain NIVMTQSPATLSLSPGERVTLSCRASEIVGTYVSWFQQKPGQ APRLLIYGASNRATGIPARFSGSGSGTDFTLTISSLQPEDLADYYC GQSYNFPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVC LLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 77) Amino acid sequence of 6F7 H3L3(hG4) heavy chain QVQLVQSGAEVVKPGASVKVSCKASGYTFTSYWMNWVRQA PGQGLEWIGIIDPSDSETSYAQKFQGRVTLTVDKSTSTAYMELSSL RSEDTAVYYCARLYRWYFDVWGAGTTVTVSSASTKGPSVFPLAP CSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYG PPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQE DPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSL GK (SEQ ID NO: 78) Amino acid sequence of 6F7 H3L3(hG4) light chain NIVMTQSPATLSLSPGERVTLSCRASEIVGTYLSWYQQKPGQ APRLLIYGASTRATGIPARFSGSGSGTDFTLTISSLQPEDFAVYYC GQSYNFPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVC LLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 79) Experimental Example 1: Phagocytosis of Tumor Cells by Macrophages Promoted by Administration of Anti-LAG3-Anti-PD-1 Bispecific Antibody in Combination with Anti-CD47 Monoclonal Antibody Macrophage induction: PBMCs (from a healthy donor) were cultured overnight, and then the cells were collected; monocytes were isolated using a Pan Monocyte Isolation Kit (Miltenyi, Cat: 130-096-537) according to the manual isolation instructions, and M-CSF (peprotech, Cat: 300-25) at a final concentration of 100 ng / mL was added; the cells were cultured in a 37 °C, 5% CO2 incubator; half of the medium was exchanged and M-CSF at a final concentration of 100 ng / mL was added on day 3, half of the medium was exchanged and M-CSF at a final concentration of 150 ng / mL was added on day 5, and the culture was continued for 2 days; and then the induced macrophages were collected. Treatment of Raji cells with IFN-y: On day 5 of macrophage induction, Raji cells (purchased from the Chinese Academy of Sciences) were conventionally passaged and treated with IFN-y (sinobiological, Cat: 11725-HNAS-100, at a final concentration of 100 ng / mL), and the cells were cultured in an incubator for 2 days. CFSE staining treatment of Raji cells: On the day of the experiment, the Raji cells that had been treated with IFN-y for 2 days were conventionally collected and counted; CFSE (Biolegend, Cat: 423801) was diluted with serum-free RPMI-1640 to a final concentration of 0.25 gM for staining and labeling; the cells were incubated in a 37 °C, 5% CO 2 incubator for labeling for 15 min and centrifuged; after the supernatant was discarded, the cells were resuspended in 10 mL of RPMI-1640 containing 10% FBS; and the cells were incubated in a 37 °C, 5% CO2 incubator for 10 min to restore the state, centrifuged, resuspended, and counted. According to the groups (the negative control was macrophages + Raji cells; the isotype control was the negative control + hIgG4 at a final concentration of 10 nM; the Bs-PL022B group was the negative control + Bs-PL022B at a final concentration of 30 nM; the 6F7 H1L1(hG4) group was the negative control + 6F7 H1L1(hG4) at a final concentration of 1 / 10 nM; and the 6F7 H1L1(hG4) + Bs-PL022B group was the negative control + 6F7 H1L1(hG4) at a final concentration of 1 / 10 nM + Bs-PL022B at a final concentration of 30 nM), macrophages and CFSE-labeled Raji cells that had been treated with IFN-y for 2 days were plated in a V-bottom 96-well plate, and different antibodies were added. The cells were cultured in a 37 °C, 5% CO2 incubator for 2 h (Raji cells:macrophages = 3:1). After washing, APC anti-mouse / human CD11b (BioLegend, Cat: 101212) diluted at 1:100 was added at 100 gL / well, and the mixture was incubated on ice for 30 min in the dark. After washing, 200 gL of 1% PBSA was added to resuspend the cells, and then the phagocytosis rate (CD11b+CFSE+%) was detected by flow cytometry. The results are shown in FIG. 1. The results show that compared with the isotype control (hIgG4), the anti-CD47 monoclonal antibody 6F7 H1L1(hG4) and the anti-LAG3-anti-PD-1 bispecific antibody Bs-PL022B in combination with the anti-CD47 monoclonal antibody 6F7 H1L1(hG4) can both effectively promote phagocytosis of IFN—Y—treated Raji cells by macrophages, and the anti-LAG3-anti-PD-1 bispecific antibody in combination with the anti-CD47 monoclonal antibody has stronger pro-phagocytic activity than the anti-CD47 monoclonal antibody. Although the specific embodiments of the present disclosure have been described in detail, those skilled in the art will understand that various modifications and substitutions can be made to those details according to all the teachings that have been disclosed, and these changes shall all fall within the protection scope of the present disclosure. The full scope of the present disclosure is given by the appended claims and any equivalent thereof.

Claims

1. A therapeutic combination, comprising an effective amount of a first antibody and an effective amount of a second antibody, wherein the first antibody is an anti-LAG3-anti-PD-1 bispecific antibody, and the second antibody is an anti-CD47 antibody or an antigen-binding fragment thereof.

2. The therapeutic combination according to claim 1, wherein:the anti-LAG3-anti-PD-1 bispecific antibody comprises:a first protein functional region targeting LAG3, anda second protein functional region targeting PD-1;whereinthe first protein functional region comprises a heavy chain variable region VH1 and a light chain variable region VL1, wherein the VH1 comprises an HCDR1 set forth in SEQ ID NO: 5, an HCDR2 set forth in SEQ ID NO: 6, and an HCDR3 set forth in SEQ ID NO: 7; and the VL1 comprises an LCDR1 set forth in SEQ ID NO: 8, an LCDR2 set forth in SEQ ID NO: 9, and an LCDR3 set forth in SEQ ID NO: 10, or the VL1 comprises an LCDR1 set forth in SEQ ID NO: 8, an LCDR2 set forth in SEQ ID NO: 46, and an LCDR3 set forth in SEQ ID NO: 47, or the VL1 comprises an LCDR1 set forth in SEQ ID NO: 48, an LCDR2 set forth in SEQ ID NO: 46, and an LCDR3 set forth in SEQ ID NO: 10;the second protein functional region comprises a heavy chain variable region VH2 and a light chain variable region VL2, wherein the VH2 comprises an HCDR1 set forth in SEQ ID NO: 26, an HCDR2 set forth in SEQ ID NO: 27, and an HCDR3 set forth in SEQ ID NO: 28; and the VL2 comprises an LCDR1 set forth in SEQ ID NO: 29, an LCDR2 setforth in SEQ ID NO: 30, and an LCDR3 set forth in SEQ ID NO: 31; and the anti-CD47 antibody or the antigen-binding fragment thereofcomprises a heavy chain variable region VH3 and a light chain variable region VL3, wherein the VH3 comprises an HCDR1 set forth in SEQ ID NO: 53, an HCDR2 set forth in SEQ ID NO: 54, and an HCDR3 set forth in SEQ ID NO: 55; and the VL3 comprises an LCDR1 set forth in SEQ ID NO: 56, an LCDR2 set forth in SEQ ID NO: 57, and an LCDR3 set forth in SEQ ID NO: 58.

3. The therapeutic combination according to claim 2, wherein:the amino acid sequence of the VH1 is set forth in SEQ ID NO: 2, and the amino acid sequence of the VL1 is selected from SEQ ID NO: 4, SEQ ID NO: 42, and SEQ ID NO: 44;the amino acid sequence of the VH2 is selected from SEQ ID NO: 15 and SEQ ID NO: 19, and the amino acid sequence of the VL2 is selected from SEQ ID NO: 17, SEQ ID NO: 21, and SEQ ID NO: 38; and / orthe amino acid sequence of the VH3 is selected from SEQ ID NO: 50, SEQ ID NO: 60, SEQ ID NO: 64, and SEQ ID NO: 68, and the amino acid sequence of the VL3 is selected from SEQ ID NO: 52, SEQ ID NO: 62, SEQ ID NO: 66, and SEQ ID NO: 70.

4. The therapeutic combination according to any one of claims 2 to 3, wherein:(1) the amino acid sequence of the VH1 is set forth in SEQ ID NO: 2, and the amino acid sequence of the VL1 is set forth in SEQ ID NO: 4; theamino acid sequence of the VH2 is set forth in SEQ ID NO: 19, and theamino acid sequence of the VL2 is set forth in SEQ ID NO: 21; and theamino acid sequence of the VH3 is set forth in SEQ ID NO: 50, and theamino acid sequence of the VL3 is set forth in SEQ ID NO: 52;(2) the amino acid sequence of the VH1 is set forth in SEQ ID NO: 2, and the amino acid sequence of the VL1 is set forth in SEQ ID NO: 4; theamino acid sequence of the VH2 is set forth in SEQ ID NO: 19, and theamino acid sequence of the VL2 is set forth in SEQ ID NO: 21; and theamino acid sequence of the VH3 is set forth in SEQ ID NO: 60, and theamino acid sequence of the VL3 is set forth in SEQ ID NO: 62;(3) the amino acid sequence of the VH1 is set forth in SEQ ID NO: 2, and the amino acid sequence of the VL1 is set forth in SEQ ID NO: 4; theamino acid sequence of the VH2 is set forth in SEQ ID NO: 19, and theamino acid sequence of the VL2 is set forth in SEQ ID NO: 21; and theamino acid sequence of the VH3 is set forth in SEQ ID NO: 64, and theamino acid sequence of the VL3 is set forth in SEQ ID NO: 66;(4) the amino acid sequence of the VH1 is set forth in SEQ ID NO: 2, and the amino acid sequence of the VL1 is set forth in SEQ ID NO: 4; theamino acid sequence of the VH2 is set forth in SEQ ID NO: 19, and theamino acid sequence of the VL2 is set forth in SEQ ID NO: 21; and theamino acid sequence of the VH3 is set forth in SEQ ID NO: 68, and theamino acid sequence of the VL3 is set forth in SEQ ID NO: 70;(5) the amino acid sequence of the VH1 is set forth in SEQ ID NO: 2, and the amino acid sequence of the VL1 is set forth in SEQ ID NO: 4; theamino acid sequence of the VH2 is set forth in SEQ ID NO: 19, and theamino acid sequence of the VL2 is set forth in SEQ ID NO: 38; and theamino acid sequence of the VH3 is set forth in SEQ ID NO: 50, and theamino acid sequence of the VL3 is set forth in SEQ ID NO: 52;(6) the amino acid sequence of the VH1 is set forth in SEQ ID NO: 2, and the amino acid sequence of the VL1 is set forth in SEQ ID NO: 4; theamino acid sequence of the VH2 is set forth in SEQ ID NO: 19, and theamino acid sequence of the VL2 is set forth in SEQ ID NO: 38; and theamino acid sequence of the VH3 is set forth in SEQ ID NO: 60, and theamino acid sequence of the VL3 is set forth in SEQ ID NO: 62;(7) the amino acid sequence of the VH1 is set forth in SEQ ID NO: 2, and the amino acid sequence of the VL1 is set forth in SEQ ID NO: 4; theamino acid sequence of the VH2 is set forth in SEQ ID NO: 19, and theamino acid sequence of the VL2 is set forth in SEQ ID NO: 38; and theamino acid sequence of the VH3 is set forth in SEQ ID NO: 64, and theamino acid sequence of the VL3 is set forth in SEQ ID NO: 66;or(8) the amino acid sequence of the VH1 is set forth in SEQ ID NO: 2, and the amino acid sequence of the VL1 is set forth in SEQ ID NO: 4; theamino acid sequence of the VH2 is set forth in SEQ ID NO: 19, and theamino acid sequence of the VL2 is set forth in SEQ ID NO: 38; and theamino acid sequence of the VH3 is set forth in SEQ ID NO: 68, and theamino acid sequence of the VL3 is set forth in SEQ ID NO: 70.

5. The therapeutic combination according to any one of claims 1 to 4, wherein:the first protein functional region and the second protein functional region are each independently selected from an immunoglobulin and a single-chain variable fragment;the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain variable fragment; orthe first protein functional region is a single-chain variable fragment, and the second protein functional region is an immunoglobulin;preferably, the C-terminus of each of two heavy chains of the immunoglobulin is linked to one single-chain variable fragment.

6. The therapeutic combination according to claim 5, wherein the therapeutic combination is characterized by any one or more of thefollowing:(1) the immunoglobulin comprises a non-CDR region derived from a human antibody;(2) the immunoglobulin comprises a constant region, wherein the constant region is derived from a human antibody; preferably, the constant region is selected from constant regions of human IgG1, IgG2, IgG3, and IgG4;(3) the immunoglobulin comprises a heavy chain constant region, and the heavy chain constant region is Ig gamma-1 chain C region (e.g., as set forth in SEQ ID NO: 39) or Ig gamma-4 chain C region (e.g., as set forth in SEQ ID NO: 45 or SEQ ID NO: 71); andthe immunoglobulin comprises a light chain constant region, and the light chain constant region is Ig kappa chain C region (e.g., as set forth in SEQ ID NO: 40);(4) the immunoglobulin is of human IgG1 subtype,the immunoglobulin comprises a heavy chain constant region, wherein, according to the EU numbering system, the heavy chain constant region comprises the following mutations:L234A and L235A;L234A and G237A;L235A and G237A;orL234A, L235A, and G237A;(5) the first protein functional region is an immunoglobulin, comprising a heavy chain having the amino acid sequence set forth in SEQ ID NO: 11, and a light chain having the amino acid sequence set forth in SEQ ID NO: 12;(6) the second protein functional region is an immunoglobulin, comprising a heavy chain having the amino acid sequence set forth inSEQ ID NO: 34, and a light chain having the amino acid sequence set forth in SEQ ID NO: 25;(7) the immunoglobulin is of human IgG4 subtype,the immunoglobulin comprises a heavy chain constant region, wherein, according to the EU numbering system, the heavy chain constant region comprises the following mutations:F234A and L235A;F234A and G237A;L235A and G237A;orF234A, L235A, and G237A;(8) the first protein functional region is an immunoglobulin, comprising a heavy chain having the amino acid sequence set forth in SEQ ID NO: 13, and a light chain having the amino acid sequence set forth in SEQ ID NO: 12;(9) the second protein functional region is an immunoglobulin, comprising a heavy chain having the amino acid sequence set forth in SEQ ID NO: 32, and a light chain having the amino acid sequence set forth in SEQ ID NO: 25.

7. The therapeutic combination according to any one of claims 5 to 6, wherein the therapeutic combination is characterized by any one or more of the following:(1) the anti-CD47 antibody comprises a non-CDR region derived from a human antibody;(2) the anti-CD47 antibody comprises a constant region, wherein the constant region is derived from a human antibody; preferably, the constant region is selected from constant regions of human IgG1, IgG2, IgG3, and IgG4;(3) the anti-CD47 antibody comprises a heavy chain constant region, and the heavy chain constant region is Ig gamma-1 chain C region (e.g., as set forth in SEQ ID NO: 39) or Ig gamma-4 chain C region (e.g., as set forth in SEQ ID NO: 45 or SEQ ID NO: 71); andthe anti-CD47 antibody comprises a light chain constant region, and the light chain constant region is Ig kappa chain C region (e.g., as set forth in SEQ ID NO: 40);(4) the anti-CD47 antibody is of human IgG1 subtype,the anti-CD47 antibody comprises a heavy chain constant region, wherein, according to the EU numbering system, the heavy chain constant region comprises the following mutations:L234A and L235A;L234A and G237A;L235A and G237A;orL234A, L235A, and G237A;(5) the anti-CD47 antibody is of human IgG4 subtype,the anti-CD47 antibody comprises a heavy chain constant region, wherein, according to the EU numbering system, the heavy chain constant region comprises the following mutations:F234A and L235A;F234A and G237A;L235A and G237A;orF234A, L235A, and G237A;(6) the anti-CD47 antibody comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 74, and a light chain having the amino acid sequence set forth in SEQ ID NO: 75;the anti-CD47 antibody comprises a heavy chain having the aminoacid sequence set forth in SEQ ID NO: 76, and a light chain having the amino acid sequence set forth in SEQ ID NO: 77; orthe anti-CD47 antibody comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 78, and a light chain having the amino acid sequence set forth in SEQ ID NO: 79;(7) the anti-CD47 antibody or the antigen-binding fragment thereof is selected from Fab, Fab', F(ab')2, Fd, Fv, dAb, a complementarity determining region fragment, a single-chain variable fragment, a humanized antibody, a chimeric antibody, and a diabody.

8. The therapeutic combination according to any one of claims 1 to 7, wherein the therapeutic combination is characterized by any one or more of the following:(1) the first protein functional region and the second protein functional region are linked directly or via a linker;preferably, the linker is a polypeptide set forth in SEQ ID NO: 73 or a polypeptide formed by concatenating a plurality of (e.g., 2, 3, 4, 5, or 6) polypeptides set forth in SEQ ID NO: 73;preferably, the linker is a polypeptide having one or more glycines linked to the C-terminus of the polypeptide set forth in SEQ ID NO: 73 or a polypeptide formed by concatenating a plurality of (e.g., 2, 3, 4, 5, or 6) polypeptides set forth in SEQ ID NO: 73;(2) the first protein functional region and the second protein functional region are each independently 1, 2, or more in number;(3) the anti-LAG3-anti-PD-1 bispecific antibody is of the IgG-scFv form.

9. The therapeutic combination according to any one of claims 1 to 8, whereinthe anti-LAG3-anti-PD-1 bispecific antibody comprises:a first protein functional region targeting LAG3, anda second protein functional region targeting PD-1;the first protein functional region is 1 in number, and the second protein functional region is 2 in number;wherein the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain variable fragment;the immunoglobulin comprises a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 2, and a light chain variable region having the amino acid sequence set forth in SEQ ID NO: 4;the single-chain variable fragment comprises a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 19, and a light chain variable region having the amino acid sequence set forth in SEQ ID NO: 21 or SEQ ID NO: 38;the single-chain variable fragment is linked to the C-termini of the two heavy chains of the immunoglobulin;the first protein functional region and the second protein functional region are linked via a first linker; the heavy chain variable region of the single-chain variable fragment and the light chain variable region of the single-chain variable fragment are linked via a second linker; the first linker and the second linker are identical or different;preferably, the amino acid sequences of the first linker and the second linker are each independently selected from SEQ ID NOs: 35-37;preferably, the amino acid sequences of the first linker and the second linker are each set forth in SEQ ID NO: 36.

10. The therapeutic combination according to any one of claims 1 to9, whereinthe anti-LAG3-anti-PD-1 bispecific antibody comprises:a first protein functional region targeting LAG3, anda second protein functional region targeting PD-1;the first protein functional region is 1 in number, and the second protein functional region is 2 in number;wherein the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain variable fragment;the immunoglobulin comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 11 or SEQ ID NO: 13, and a light chain having the amino acid sequence set forth in SEQ ID NO: 12;the single-chain variable fragment comprises a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 19, and a light chain variable region having the amino acid sequence set forth in SEQ ID NO: 21 or SEQ ID NO: 38;the single-chain variable fragment is linked to the C-termini of the two heavy chains of the immunoglobulin;the first protein functional region and the second protein functional region are linked via a first linker; the heavy chain variable region of the single-chain variable fragment and the light chain variable region of the single-chain variable fragment are linked via a second linker; the first linker and the second linker are identical or different;preferably, the amino acid sequences of the first linker and the second linker are each independently selected from SEQ ID NOs: 35-37;preferably, the amino acid sequences of the first linker and the second linker are each set forth in SEQ ID NO: 36.

11. The therapeutic combination according to any one of claims 1 to10, whereinthe anti-LAG3-anti-PD-1 bispecific antibody comprises:a first protein functional region targeting LAG3, anda second protein functional region targeting PD-1;the first protein functional region is 1 in number, and the second protein functional region is 2 in number;wherein the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain variable fragment;the immunoglobulin comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 11, and a light chain having the amino acid sequence set forth in SEQ ID NO: 12;the single-chain variable fragment comprises a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 19, and a light chain variable region having the amino acid sequence set forth in SEQ ID NO: 38;the single-chain variable fragment is linked to the C-termini of the two heavy chains of the immunoglobulin;the first protein functional region and the second protein functional region are linked via a first linker; the heavy chain variable region of the single-chain variable fragment and the light chain variable region of the single-chain variable fragment are linked via a second linker; the first linker and the second linker are identical or different;preferably, the amino acid sequences of the first linker and the second linker are each independently selected from SEQ ID NOs: 35-37;preferably, the amino acid sequences of the first linker and the second linker are each set forth in SEQ ID NO: 36.

12. The therapeutic combination according to any one of claims 1 to8, whereinthe anti-LAG3-anti-PD-1 bispecific antibody comprises:a first protein functional region targeting LAG3, anda second protein functional region targeting PD-1;the first protein functional region is 2 in number, and the second protein functional region is 1 in number;wherein the first protein functional region is a single-chain variable fragment, and the second protein functional region is an immunoglobulin;the single-chain variable fragment comprises a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 2, and a light chain variable region having the amino acid sequence set forth in SEQ ID NO: 4;the immunoglobulin comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 34 or SEQ ID NO: 32, and a light chain having the amino acid sequence set forth in SEQ ID NO: 25;the single-chain variable fragment is linked to the C-termini of the two heavy chains of the immunoglobulin;the first protein functional region and the second protein functional region are linked via a third linker, and the heavy chain variable region of the single-chain variable fragment and the light chain variable region of the single-chain variable fragment are linked via a fourth linker; the third linker and the fourth linker are identical or different;preferably, the amino acid sequences of the third linker and the fourth linker are each independently selected from SEQ ID NOs: 35-37;preferably, the amino acid sequences of the third linker and the fourth linker are each set forth in SEQ ID NO: 36.

13. The therapeutic combination according to any one of claims 1 to 11, wherein the anti-LAG3-anti-PD-1 bispecific antibody comprises:a first protein functional region targeting LAG3, anda second protein functional region targeting PD-1;the first protein functional region is 1 in number, and the second protein functional region is 2 in number;wherein the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain variable fragment;the immunoglobulin comprises a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 2, and a light chain variable region having the amino acid sequence set forth in SEQ ID NO: 4;the single-chain variable fragment comprises a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 19, and a light chain variable region having the amino acid sequence set forth in SEQ ID NO: 21 or SEQ ID NO: 38;the single-chain variable fragment is linked to the C-termini of the two heavy chains of the immunoglobulin;the first protein functional region and the second protein functional region are linked via a first linker; the heavy chain variable region of the single-chain variable fragment and the light chain variable region of the single-chain variable fragment are linked via a second linker; the first linker and the second linker are identical or different;preferably, the amino acid sequences of the first linker and the second linker are each independently selected from SEQ ID NOs: 35-37;preferably, the amino acid sequences of the first linker and the second linker are each set forth in SEQ ID NO: 36;the anti-CD47 antibody comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 74, and a light chain having the amino acid sequence set forth in SEQ ID NO: 75.

14. The therapeutic combination according to any one of claims 1 to 13, wherein the therapeutic combination is characterized by any one or more of the following:(1) the first antibody and the second antibody are mixed together, and the therapeutic combination is a pharmaceutical composition;(2) the first antibody and the second antibody are in separate packages, and the therapeutic combination is a combination product;(3) the mass ratio of the first antibody to the second antibody is (10:1) to (1:5), (5:1) to (1:3), (3:1) to (1:2), or 2:1;(4) the unit dose of the first antibody and the second antibody is independently 100 mg-2500 mg, 100 mg-2000 mg, 100 mg-1500 mg, 100 mg-1200 mg, 100 mg-1000 mg, 200 mg-800 mg, 200 mg-500 mg, 300 mg-600 mg, 400 mg-500 mg, or 450 mg, based on the mass of the first antibody or the second antibody;(5) the therapeutic combination further comprises an effective amount of an anti-tumor chemotherapeutic drug, such as an alkylating agent, an antimetabolite, an anti-tumor antibiotic, a plant-based anticancer agent, a hormone, or an immunological agent;(6) the therapeutic combination further comprises one or more pharmaceutically acceptable auxiliary materials; preferably, further comprises a package insert;(7) the active pharmaceutical ingredient of the therapeutic combination consists of the first antibody and the second antibody.

15. The therapeutic combination according to any one of claims 1 to 14 for use in treating or preventing a tumor, whereinpreferably, the tumor is selected from one or more of pancreatic cancer, breast cancer, ovarian cancer, colorectal cancer, head and neckcancer, brain cancer, throat cancer, nasopharyngeal cancer, thyroid cancer, mesothelioma, lung cancer, liver cancer, gastric cancer, biliary tract cancer, renal cancer, fallopian tube cancer, endometrial cancer, cervical cancer, bladder cancer, urothelial carcinoma, prostate cancer, testicular cancer, skin cancer, melanoma, myeloma, plasma cell cancer, vulval cancer, leukemia, lymphoma, bone cancer, esophageal cancer, and osteosarcoma;preferably, the lung cancer is selected from one or more of non-small cell lung cancer, small cell lung cancer and squamous cell lung cancer;preferably, the gastric cancer is gastric adenocarcinoma or gastroesophageal junction adenocarcinoma;preferably, the lymphoma is Hodgkin’s lymphoma or nonHodgkin’s lymphoma;preferably, the esophageal cancer is esophageal squamous cell carcinoma.

16. The therapeutic combination according to any one of claims 1 to 15, wherein the therapeutic combination is characterized by any one or more of the following:(1) the first antibody and the second antibody are independently administered before or after surgery and / or before or after radiotherapy;(2) the unit dose of the first antibody and the second antibody is independently 0.1-100 mg, 5-50 mg, 1-10 mg, or 5-15 mg per kg body weight; or the unit dose of the first antibody and the second antibody is independently 10-1000 mg, 50-500 mg, 100-400 mg, 150-300 mg, 150-250 mg, or 200 mg in each subject;(3) the first antibody and the second antibody are each independently administered once every half day, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 10 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks,or 6 weeks;(4) the route of administration is intravenous drip infusion, intravenous injection, subcutaneous injection, or intraperitoneal injection.

17. Use of the therapeutic combination according to any one of claims 1 to 14 in the preparation of a medicament for treating or preventing a tumor, whereinpreferably, the tumor is selected from one or more of pancreatic cancer, breast cancer, ovarian cancer, colorectal cancer, head and neck cancer, brain cancer, throat cancer, nasopharyngeal cancer, thyroid cancer, mesothelioma, lung cancer, liver cancer, gastric cancer, biliary tract cancer, renal cancer, fallopian tube cancer, endometrial cancer, cervical cancer, bladder cancer, urothelial carcinoma, prostate cancer, testicular cancer, skin cancer, melanoma, myeloma, plasma cell cancer, vulval cancer, leukemia, lymphoma, bone cancer, esophageal cancer, and osteosarcoma;preferably, the lung cancer is selected from one or more of non-small cell lung cancer, small cell lung cancer and squamous cell lung cancer;preferably, the gastric cancer is gastric adenocarcinoma or gastroesophageal junction adenocarcinoma;preferably, the lymphoma is Hodgkin’s lymphoma or nonHodgkin’s lymphoma;preferably, the esophageal cancer is esophageal squamous cell carcinoma.

18. The use according to claim 17, wherein the use is characterized by any one or more of the following:(1) the first antibody and the second antibody are independentlyadministered before or after surgery and / or before or after radiotherapy;(2) the unit dose of the first antibody and the second antibody is independently 0.1-100 mg, 5-50 mg, 1-10 mg, or 5-15 mg per kg body weight; or the unit dose of the first antibody and the second antibody is independently 10-1000 mg, 50-500 mg, 100-400 mg, 150-300 mg, 150-250 mg, or 200 mg in each subject;(3) the first antibody and the second antibody are each independently administered once every half day, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 10 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, or 6 weeks;(4) the route of administration is intravenous drip infusion, intravenous injection, subcutaneous injection, or intraperitoneal injection.

19. A method for treating or preventing a tumor, comprising a step of administering to a subject in need an effective amount of the therapeutic combination according to any one of claims 1 to 14, whereinpreferably, the tumor is selected from one or more of pancreatic cancer, breast cancer, ovarian cancer, colorectal cancer, head and neck cancer, brain cancer, throat cancer, nasopharyngeal cancer, thyroid cancer, mesothelioma, lung cancer, liver cancer, gastric cancer, biliary tract cancer, renal cancer, fallopian tube cancer, endometrial cancer, cervical cancer, bladder cancer, urothelial carcinoma, prostate cancer, testicular cancer, skin cancer, melanoma, myeloma, plasma cell cancer, vulval cancer, leukemia, lymphoma, bone cancer, esophageal cancer, and osteosarcoma;preferably, the lung cancer is selected from one or more of non-small cell lung cancer, small cell lung cancer and squamous cell lung cancer;preferably, the gastric cancer is gastric adenocarcinoma orgastroesophageal junction adenocarcinoma;preferably, the lymphoma is Hodgkin’s lymphoma or nonHodgkin’s lymphoma;preferably, the esophageal cancer is esophageal squamous cell carcinoma.

20. The method for treating or preventing a tumor according to claim 19, wherein the method is characterized by any one or more of the following:(1) the first antibody and the second antibody are independently administered before or after surgery and / or before or after radiotherapy;(2) the unit dose of the first antibody and the second antibody is independently 0.1-100 mg, 5-50 mg, 1-10 mg, or 5-15 mg per kg body weight; or the unit dose of the first antibody and the second antibody is independently 10-1000 mg, 50-500 mg, 100-400 mg, 150-300 mg, 150-250 mg, or 200 mg in each subject;(3) the first antibody and the second antibody are each independently administered once every half day, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 10 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, or 6 weeks;(4) the route of administration is intravenous drip infusion, intravenous injection, subcutaneous injection, or intraperitoneal injection.

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