Bispecific antibody capable of being combined with immune cells to enhance tumor killing capability, and preparation method therefor and application thereof

Abstract

The present invention provides a bispecific antibody capable of being combined with immune cells to enhance a targeting tumor killing capability, and a preparation method therefor and an application thereof. Antibodies and degradable nanoparticles are connected by using a chemical method, so as to make the one nanoparticle be connected to two or more antibody molecules at the same time, wherein one antibody can be specifically bound with immune cells, and the other antibody or the other antibodies can be specifically bound to tumor cells so as to achieve the effect of enhancing the capability of the immune cells for specifically killing tumor cells in a targeting way.

1 .- 17 . (canceled) 
     
     
         18 . A bispecific antibody, comprising a first antibody moiety that binds to an antigen expressed on an effector T cell and a second antibody moiety that binds to an antigen expressed on a cancer cell, wherein the first antibody moiety and the second antibody moiety are connected by a biodegradable nanomaterial, wherein the antigen expressed on an effector T cell is not CD40. 
     
     
         19 . The bispecific antibody according to  claim 18 , wherein the nanomaterial is any one of polylactic acid-glycolic acid, poly lactic acid, polycaprolactone, polybutylene glycol succinate, polyaniline, polycarbonate, glycolide-lactide copolymer or glycolide-caprolactone copolymer, or a mixture thereof. 
     
     
         20 . The bispecific antibody according to  claim 18 , wherein the antigen expressed on the cancer cell is selected from the group consisting of CD19, CD20, CD22, CD30, CD33, CD123, Muc1, Muc16, HER2, HER3, EGFRvIII, VEGFA, CEA, GPA33, GP100, ANG2, L1CAM, ROR-1, CS1, MICA and MICB; the antigen expressed on the effector T cell is selected from the group consisting of CD2, CD3, CD4, CD5, CD6, CD8, CD25, CD28, CD30, CD40L, CD44, CD45, CD69, and CD90. 
     
     
         21 . The bispecific antibody according to  claim 18 , wherein the antigen expressed on the cancer cell is selected from the group consisting of carbonic anhydrase IX, alpha-fetoprotein, alpha-actinin-4, A3, A33 antibody-specific antigen, ANG2, ART-4, B7, Ba 733, BAGE, BrE3-antigen, CA125, CAMEL, CAP-1, CASP-8/m, CCCL19, CCCL21, CD1, CD1a, CD2, CD3, CD4, CD5, CD8, CD11A, CD14, CD15, CD16, CD18, CD19, CD20, CD21, CD22, CD23, CD25, CD29, CD30, CD32b, CD33, CD37, CD38, CD40, CD40L, CD45, CD46, CD52, CD54, CD55, CD59, CD64, CD66a-e, CD67, CD70, CD70L, CD74, CD79a, CD79b, CD80, CD83, CD95, CD126, CD132, CD133, CD138, CD147, CD154, CDC27, CDK-4/m, CDKN2A, CS1, CXCR4, CXCR7, CXCL12, HIF-1alpha, colon-specific antigen-p (CSAp), CEA(CEACAM5), CEACAM6, c-met, DAM, EGFR, EGFRvIII, EGP-1, EGP-2, ELF2-M, Ep-CAM, Flt-1, Flt-3, folate receptor, G250 antigen, GAGE, gp100, GPA33, GROB, HLA-DR, HM1.24, human chorionic gonadotropin (HCG) and its subunit, HER2/neu, HERS, HMGB-1, hypoxia-inducible factor (HIF-1), HSP70-2M, HST-2, Ia, IGF-1R, IFN-γ, IFN-α, IFN-β, IL-2, IL-4R, IL-6R, IL-13R, IL-15R, IL-17R, IL-18R, IL-6, IL-8, IL-12, IL-15, IL-17, IL-18, IL-23, IL-25, IL-33, insulin-like growth factor-1 (IGF-1), KC4-antigen, KS-1-antigen, KS1-4, L1CAM, Le-Y, LDR/FUT, macrophage migration inhibitory factor (MIF), MAGE, MAGE-3, MART-1, MART-2, NY-ESO-1, TRAG-3, mCRP, MCP-1, MICA, MICB, MIP-1A, MIP-1B, MIF, MUC1, MUC2, MUC3, MUC4, MUC5ac, MUC13, MUC16, MUM-1/2, MUM-3, NCA66, NCA95, NCA90, pancreatic cancer mucin, placental growth factor, p53, PLAGL2, prostate acid phosphatase, PSA, PRAME, PSMA, P1GF, ILGF, ILGF-1R, IL-6, IL-25, ROR-1, RS5, RANTES, T101, SAGE, S100, survivin, survivin-2B, TAC, TAG-72, tenascin, TRAIL receptor, TNF-alpha, Tn antigen, Thomson-Friedrich antigen, tumor necrosis antigen, TROP-2, VEGFA, VEGFR, ED-B fibronectin, WT-1, 17-1A-antigen, complement factor C3, C3a, C3b, C5a, C5, angiogenesis marker, bc1-2, bc1-6, Kras, and cMET, and the antigen expressed on the effector T cell is selected from the group consisting of ADAM17, CD2, CD3, CD4, CD5, CD6, CD8, CD11a, CD11b, CD14, CD16, CD16b, CD25, CD28, CD30, CD32a, CD40L, CD44, CD45, CD56, CD57, CD64, CD69, CD74, CD89, CD90, CD137, CD177, CEACAM6, CEACAM8, HLA-DRa chain, KIR, LSECtin or SLC44A2. 
     
     
         22 . A bispecific antibody, comprising a first antibody moiety that binds to an antigen expressed on an effector T cell and a second antibody moiety that binds to an antigen expressed on a cancer cell, wherein the first antibody moiety and the second antibody moiety are connected by a biodegradable nanomaterial, and
 wherein the antigen expressed on the effector T cell is CD3 and the antigen expressed on the target cell is Muc1, or   wherein the antigen expressed on the effector T cell is CD3 and the antigen expressed on the target cell is CD19, or   wherein the antigen expressed on the effector T cell is CD3 and the antigen expressed on the target cell is CD20, or   wherein the antigen expressed on the effector T cell is CD3 and the antigen expressed on the target cell is CD33, or   wherein the antigen expressed on the effector T cell is CD3 and the antigen expressed on the target cell is Her2, or   wherein the antigen expressed on the effector T cell is CD8 and the antigen expressed on the target cell is Muc1, or   wherein the antigen expressed on the effector T cell is CD3 and the antigen expressed on the target cell is CD38.   
     
     
         23 . The bispecific antibody according to  claim 22 , wherein the nanomaterial is any one of polylactic acid-glycolic acid, polylactic acid, polycaprolactone, polybutylene glycol succinate, polyaniline, polycarbonate, glycolide-lactide copolymer or glycolide-caprolactone copolymer, or a mixture thereof. 
     
     
         24 . A method for producing a bispecific antibody according to  claim 18 , which comprises connecting the nanomaterial to the first antibody moiety and to the second antibody moiety. 
     
     
         25 . The method according to  claim 24 , which comprises the steps of: (1) preparation, collection and activation of the nanomaterial; (2) connecting the nanomaterial obtained in step (1) with a mixture of the first antibody moiety and the second antibody moiety. 
     
     
         26 . The method according to  claim 25 , wherein the nanomaterial is any one of polylactic acid-glycolic acid, poly lactic acid, polycaprolactone, polybutylene glycol succinate, polyaniline, polycarbonate, glycolide-lactide copolymer or glycolide-caprolactone copolymer, or a mixture thereof; and the nanomaterial dissolved in a solvent selected from acetone, butanone, methanol, ethanol or isopropanol or a mixture thereof. 
     
     
         27 . A method of treating a tumor in a subject comprising administering to the subject a bispecific antibody according to  claim 18 . 
     
     
         28 . The method according to  claim 27 , wherein the tumor is selected from the group consisting of liver cancer, non-small cell lung cancer, small cell lung cancer, adrenocortical carcinoma, acute (chronic B) lymphocytoma, myeloma, prostate cancer, breast cancer, esophageal cancer, gastric cancer, colorectal cancer, cervical cancer, kidney cancer, bladder cancer and lymphoma. 
     
     
         29 . A method for producing a bispecific antibody according to  claim 22 , which comprises connecting the nanomaterial to the first antibody moiety and to the second antibody moiety. 
     
     
         30 . The method according to  claim 29 , which comprises the steps of: (1) preparation, collection and activation of the nanomaterial; (2) connecting the nanomaterial obtained in step (1) with a mixture of the first antibody moiety and the second antibody moiety. 
     
     
         31 . The method according to  claim 30 , wherein the nanomaterial is any one of polylactic acid-glycolic acid, poly lactic acid, polycaprolactone, polybutylene glycol succinate, polyaniline, polycarbonate, glycolide-lactide copolymer or glycolide-caprolactone copolymer, or a mixture thereof; and the nanomaterial dissolved in a solvent selected from acetone, butanone, methanol, ethanol or isopropanol or a mixture thereof. 
     
     
         32 . A method of treating a tumor in a subject comprising administering to the subject a bispecific antibody according to  claim 22 . 
     
     
         33 . The method according to  claim 32 , wherein the tumor is selected from the group consisting of liver cancer, non-small cell lung cancer, small cell lung cancer, adrenocortical carcinoma, acute (chronic B) lymphocytoma, myeloma, prostate cancer, breast cancer, esophageal cancer, gastric cancer, colorectal cancer, cervical cancer, kidney cancer, bladder cancer and lymphoma.