Vegf-like factor antibodies and methods of use thereof

Abstract

A novel human gene having a significant homology with a VEGF-C gene, a member of the VEGF family, has been isolated by the PCR method using primers designed based on the sequence of EST that is assumed to be homologous with the C-terminal region of the VEGF-C gene. Mouse and rat genes have been isolated based on the human gene isolated as above. A protein encoded by the above human gene has been isolated by introducing the gene into Escherichia coli and expressing it. The isolated protein and genes can be applied to, for example, gene therapy for the VEGF-D deficiency, wound healing, and promotion of collateral vessel formation. Furthermore, VEGF-D protein inhibitors can be used as a novel anticancer drug, etc.

1 . An antibody binding to a protein shown by SEQ ID NO: 1 or having the amino acid sequence derived therefrom in which one or more amino acids are substituted, deleted, or added. 
     
     
         2 . The antibody of  claim 1 , wherein said protein is encoded by a nucleic acid hybridizing with the nucleic acid shown by SEQ ID NO: 2. 
     
     
         3 . The antibody of  claim 1 , wherein said protein is an isolated mammalian VEGF-D protein encoded by an isolated nucleic acid hybridizing under highly stringent conditions to the complement of the sequence set forth in SEQ ID NO: 2, wherein said nucleic acid has about 90% or higher sequence homology with the sequence set forth in SEQ ID NO: 2, and said highly stringent conditions involve gradient hybridization from a temperature of about 68° C. to about 55° C. for about 2 hours and washing with 2×SSC solution at room temperature, and wherein said protein has a biological activity of a VEGF family protein. 
     
     
         4 . The antibody of  claim 1 , wherein said protein is encoded by an isolated nucleic acid that is capable of hybridizing with the complement of the nucleic acid shown by SEQ ID NO: 2 under the conditions of hybridizing under a temperature gradient of 68° C. to 55° C. for 2 hours and washing with a 2×SSC solution containing 0.05% SDS at room temperature for 10 minutes for four times and with a 0.1×SSC solution containing 0.1% SDS at 45° C. for 3 minutes, and wherein said protein has at least one activity selected from the group consisting of vascular formation, lymphatic vessel formation, pathological neovascularization, increasing vascular permeability, edema formation, wound healing, and promoting collateral vessel formation. 
     
     
         5 . The antibody of  claim 1 , wherein said protein is encoded by an isolated nucleic acid having 90% or higher sequence homology to the nucleic acid shown by SEQ ID NO: 2, and wherein said protein has at least one activity selected from the group consisting of vascular formation, lymphatic vessel formation, pathological neovascularization, increasing vascular permeability, edema formation, wound healing, and promoting collateral vessel formation. 
     
     
         6 . The antibody of  claim 1 , wherein the amino acid sequence of said protein comprises the amino acid sequence of SEQ ID NO: 1. 
     
     
         7 . The antibody of  claim 1 , wherein said antibody is humanized. 
     
     
         8 . A method for inhibiting angiogenesis or neovascularization comprising administering to a mammal an effective amount of an antibody according to any one of  claims 1  to  7 . 
     
     
         9 . The method of  claim 8 , wherein the method is for treating pathological neovascularization, lymphatic dysplasia, dyshematopoiesis, or edemas.