Recombinant protein targeting pd-1 and tgfbeta

Abstract

A recombinant fusion protein comprising 1) TGFβR2, or a fragment thereof, capable of binding to TGFβ, and 2) an antibody, or an antigen-binding fragment thereof, that binds to PD-1. Also disclosed are a polynucleotide encoding the recombinant fusion protein, an expression vector containing the polynucleotide, a method for producing the recombinant protein and a method for treating a disease caused by over expression of TGFβ and/or PD-1 using the recombinant protein.

We claim: 
     
         1 . A recombinant fusion protein, comprising
 a) human TGFβR2 or a fragment thereof, that binds to TGFβ, and   b) an antibody or an antigen-binding fragment thereof comprising heavy chain variable region and light chain variable region, that binds to PD-1.   
     
     
         2 . The recombinant fusion protein according to  claim 1 , comprising:
 a) a human TGFβR2 fragment, and   b) an antibody or an antigen-binding fragment thereof binding to PD-1 and comprising 3 CDRs of a heavy chain variable region of SEQ ID NO: 2 and 3 CDRs of a light chain variable region of SEQ ID NO: 3.   
     
     
         3 . The recombinant fusion protein according to  claim 1   wherein the human TGFβR2 fragment is linked to the C-terminus of the heavy chain or an antigen-binding fragment thereof.   
     
     
         4 . The recombinant fusion protein according to  claim 1 , wherein human TGFβR2 is a TGFβR2 extracellular domain, optionally, wherein the TGFβR2 extracellular domain comprises an amino acid sequence of SEQ ID NO: 1, or an amino acid sequence having at least 90% identity with SEQ ID NO: 1. 
     
     
         5 . The recombinant fusion protein according to  claim 1 , wherein the antibody or the antigen-binding fragment thereof comprises a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 2 or an amino acid sequence having at least 90% identity with SEQ ID NO: 2 and simultaneously having 3 CDRs of SEQ ID NO: 2, and a light chain variable region comprising an amino acid sequence of SEQ ID NO: 3 or an amino acid sequence having at least 90% identity with SEQ ID NO:3 and simultaneously having 3 CDRs of SEQ ID NO:3. 
     
     
         6 . The recombinant fusion protein according to  claim 1 , wherein the antibody or the antigen-binding fragment thereof comprises a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence of SEQ ID NO: 4 or an amino acid sequence having at least 90% identity with SEQ ID NO: 4 and simultaneously having 3 CDRs of SEQ ID NO: 4, and the light chain comprises an amino acid sequence of SEQ ID NO: 5 or an amino acid sequence having at least 90% identity with SEQ ID NO:5 and simultaneously having 3 CDRs of SEQ ID NO:5. 
     
     
         7 . The recombinant fusion protein according to  claim 3 , wherein the human TGFβR2 fragment is linked via a linker to the C-terminus of the heavy chain or an antigen-binding fragment thereof, wherein the linker comprises an amino acid sequence of SEQ ID Nos.: 6, 7, 8 or 9. 
     
     
         8 . The recombinant fusion protein according to  claim 1 , selected from:
 1) a recombinant fusion protein comprising an amino acid sequence of SEQ ID NO: 10 or an amino acid sequence having at least 90% identity with SEQ ID NO: 10 and simultaneously having 3CDRs of SEQ ID NO: 4; and an amino acid sequence of SEQ ID NO: 5 or an amino acid sequence having at least 90% identity with SEQ ID NO: 5 and simultaneously having 3CDRs of SEQ ID NO: 5;   2) a recombinant fusion protein comprising an amino acid sequence of SEQ ID NO: 11 or an amino acid sequence having at least 90% identity with SEQ ID NO: 11 and simultaneously having 3CDRs of SEQ ID NO: 4, and an amino acid sequence of SEQ ID NO: 5 or an amino acid sequence having at least 90% identity with SEQ ID NO: 5 and simultaneously having 3CDRs of SEQ ID NO: 5;   3) a recombinant fusion protein comprising an amino acid sequence of SEQ ID NO: 12 or an amino acid sequence having at least 90% identity with SEQ ID NO:12 and simultaneously having 3CDRs of SEQ ID NO: 4, and an amino acid sequence of SEQ ID NO: 5 or an amino acid sequence having at least 90% identity with SEQ ID NO: 5 and simultaneously having 3CDRs of SEQ ID NO: 5; or   4) a recombinant fusion protein comprising an amino acid sequence of SEQ ID NO: 13 or an amino acid sequence having at least 90% identity with SEQ ID NO: 13 and simultaneously having 3CDRs of SEQ ID NO: 4, and an amino acid sequence of SEQ ID NO: 5 or an amino acid sequence having at least 90% identity with SEQ ID NO: 5 and simultaneously having 3CDRs of SEQ ID NO: 5.   
     
     
         9 . A nucleic acid encoding the recombinant fusion protein according to  claim 1 . 
     
     
         10 . A vector comprising the nucleic acid of  claim 9 . 
     
     
         11 . A host cell comprising the nucleic acid of  claim 9 . 
     
     
         12 . A method for producing a recombinant fusion protein, comprising:
 (1) providing a recombinant fusion protein-encoding polynucleotide molecule;   (2) constructing an expression vector comprising the polynucleotide molecule of (1);   (3) transfecting or transforming suitable host cells with the expression vector of (2) and cultivating the host cells to express the protein; and   (4) purifying the protein.   
     
     
         13 . A recombinant fusion protein produced by the method of  claim 12 . 
     
     
         14 . A pharmaceutical composition, comprising the recombinant fusion protein of  claim 1 , and at least one pharmaceutically acceptable carrier. 
     
     
         15 . A method for treating a disease over-expressing TGFβ and/or PD-L1, comprising administering to a subject in need thereof a therapeutically effective amount of the recombinant fusion protein of  claim 1 . 
     
     
         16 . The method of  claim 15 , wherein the disease is a cancer/tumor selected from the group consisting of colorectal, breast, ovarian, pancreatic, gastric, prostate, renal, cervical, myeloma, lymphoma, leukemia, thyroid, endometrial, uterine, bladder, neuroendocrine, head and neck, liver, nasopharyngeal, testicular, small cell lung cancer, non-small cell lung cancer, melanoma, basal cell skin cancer, squamous cell skin cancer, dermatofibrosarcoma protuberans, Merkel cell carcinoma, glioblastoma, glioma, sarcoma, mesothelioma, and myelodysplastic syndromes. 
     
     
         17 . A host cell comprising the vector of  claim 10 .