Bifunctional protein aiming at VEGF and TGF-beta and application of bifunctional protein
A bifunctional protein and TGF- technology, which is applied in the field of biomedicine, can solve the problems such as the news that the bifunctional protein drug has entered the clinical stage is not disclosed, and achieve the effect of improving the therapeutic effect and reducing the side effects.
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Embodiment 1
[0053] Example 1 of the present invention constructs a bifunctional protein targeting VEGF and TGF-β, including anti-VEGF monoclonal antibody capable of specifically binding to VEGF antigen, connecting peptide linker and TGF-βRII extracellular domain polypeptide, anti-VEGF The monoclonal antibody is connected to the TGF-βRII extracellular domain polypeptide through the linker peptide. The anti-VEGF monoclonal antibody includes two heavy chains and two light chains, and the TGF-βRII extracellular domain polypeptide is connected to the C-terminal or N-terminal of the two heavy chains or the two light chains of the anti-VEGF monoclonal antibody through a linker connect.
Embodiment 2
[0055] On the basis of Example 1, Example 2 of the present invention further defines the anti-VEGF monoclonal antibody. The anti-VEGF monoclonal antibody defined here is a monoclonal antibody targeting VEGF, and the present invention is preferably a bevacizumab monoclonal antibody. Antibody, the amino acid sequence of its heavy chain is SEQ ID NO:1, the amino acid sequence of its light chain is SEQ ID NO:2; the amino acid sequence of TGF-βRII extracellular domain polypeptide is SEQ ID NO:3.
[0056] The general formula of the connecting peptide linker is (GGGGS)n; the n is 4.
[0057] The bispecific antibody is selected from any of the following:
[0058] Protein EB-1, which is composed of TGF-βRII extracellular domain polypeptide connected to the C-terminals of the two heavy chains of anti-VEGF monoclonal antibody through the connecting peptide linker, the amino acid sequence of the heavy chain of protein EB-1 is SEQ ID NO :4, the amino acid sequence of its light chain is SE...
Embodiment 3
[0078] Embodiment 3 of the present invention provides a nucleotide molecule encoding a bifunctional protein targeting VEGF and TGF-β.
[0079] The present invention also provides a recombinant DNA expression vector, which contains the above-mentioned nucleotide molecules.
[0080] The present invention also provides a host cell transfected with a recombinant DNA expression vector, and the host cell includes prokaryotic, yeast or mammalian cells.
[0081] The present invention also provides the application of the bifunctional protein targeting VEGF and TGF-β in the preparation of drugs for treating tumor diseases or fibrotic diseases;
[0082] The neoplastic disease is selected from colorectal cancer, non-small cell lung cancer, breast cancer, pancreatic cancer, kidney cancer, gastric cancer, liver cancer, ovarian cancer, melanoma or glioma;
[0083] Fibrotic diseases include liver fibrosis and pulmonary fibrosis.
[0084] The present invention also provides the application o...
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