Bispecific fusion polypeptides and their applications

Abstract

The present invention provides a bispecific fusion polypeptide that binds to vascular endothelial growth factor (VEGF)A and VEGFC, the bispecific fusion polypeptide comprising a first domain that specifically recognizes VEGFA and a second domain that specifically recognizes VEGFC, and a method for treating or preventing a disease associated with VEGFA or VEGFC using the bispecific fusion polypeptide.

Claims

[Claim 1] A bispecific binding molecule for vascular endothelial growth factor (VEGF) A and VEGFC, characterized by comprising a dimer of a bispecific fusion polypeptide, wherein the bispecific fusion polypeptide comprises a first domain that binds to vascular endothelial growth factor (VEGF) A and VEGFC and specifically recognizes VEGFA, a second domain that specifically recognizes VEGFC, and a third domain, It is characterized by containing an amino acid sequence shown in any one of SEQ ID NO: 17 to 24, or an amino acid sequence having at least 98%, 99%, or 100% sequence identity with an amino acid sequence shown in any one of SEQ ID NO: 17 to 24, The first domain includes an amino acid sequence shown in SEQ ID NO:1 or an amino acid sequence having at least 98%, 99%, or 100% sequence identity with SEQ ID NO:1, and an amino acid sequence shown in SEQ ID NO:2 or an amino acid sequence having at least 98%, 99%, or 100% sequence identity with SEQ ID NO:

2. The second domain includes the amino acid sequence shown in SEQ ID NO: 10, the amino acid sequence in which the 80th amino acid corresponding to SEQ ID NO: 10 is not asparagine, or the amino acid sequence having at least 98%, 99%, or 100% sequence identity with SEQ ID NO:

10. In the amino acid sequence corresponding to SEQ ID NO: 10, where the 80th amino acid is not asparagine, the 80th amino acid is substituted with glutamine, aspartic acid, glutamic acid, arginine, or lysine. The third domain is a third domain containing the Fc region of immunoglobulin, and has the amino acid sequence shown in SEQ ID NO: 13, or an amino acid sequence having at least 98%, 99%, or 100% sequence identity with SEQ ID NO:

13. Bispecific binding molecules. [Claim 2] The bispecific binding molecule according to Claim 1, wherein the immunoglobulin is selected from IgA, IgG, IgM, IgD, and IgE. [Claim 3] The bispecific binding molecule according to claim 1, wherein the first domain, the second domain and / or the third domain are directly connected or connected via a linker. [Claim 4] The linker comprises at least six amino acids, as described in claim 3, for the bispecific linking molecule. [Claim 5] The bispecific binding molecule according to claim 4, characterized in that the linker has an amino acid sequence shown in SEQ ID NO: 7 or 8, or an amino acid sequence having one, two, or three amino acid insertions, substitutions, or deletions compared to the amino acid sequence shown in SEQ ID NO: 7 or 8. [Claim 6] The bispecific binding molecule according to claim 1, characterized in that the first domain includes an amino acid sequence shown in SEQ ID NO: 9 or an amino acid sequence having at least 98%, 99%, or 100% sequence identity with SEQ ID NO:

9. [Claim 7] The bispecific binding molecule according to claim 1, characterized in that the amino acid sequence in which the 80th amino acid corresponding to SEQ ID NO: 10 is not asparagine is the amino acid sequence shown in SEQ ID NO:

11. [Claim 8] An isolated nucleic acid molecule encoding a bispecific fusion polypeptide, the bispecific fusion polypeptide comprising: a first domain that binds to vascular endothelial growth factor (VEGF) A and VEGFC and specifically recognizes VEGFA; a second domain that specifically recognizes VEGFC; and a third domain, It is characterized by containing an amino acid sequence shown in any one of SEQ ID NO: 17 to 24, or an amino acid sequence having at least 98%, 99%, or 100% sequence identity with an amino acid sequence shown in any one of SEQ ID NO: 17 to 24, The first domain includes an amino acid sequence shown in SEQ ID NO:1 or an amino acid sequence having at least 98%, 99%, or 100% sequence identity with SEQ ID NO:1, and an amino acid sequence shown in SEQ ID NO:2 or an amino acid sequence having at least 98%, 99%, or 100% sequence identity with SEQ ID NO:

2. The second domain includes the amino acid sequence shown in SEQ ID NO: 10, the amino acid sequence in which the 80th amino acid corresponding to SEQ ID NO: 10 is not asparagine, or the amino acid sequence having at least 98%, 99%, or 100% sequence identity with SEQ ID NO:

10. In the amino acid sequence corresponding to SEQ ID NO: 10, where the 80th amino acid is not asparagine, the 80th amino acid is substituted with glutamine, aspartic acid, glutamic acid, arginine, or lysine. The third domain is a nucleic acid molecule having an amino acid sequence represented by SEQ ID NO: 13, or an amino acid sequence having at least 98%, 99%, or 100% sequence identity with SEQ ID NO:

13. [Claim 9] A nucleic acid delivery carrier comprising the isolated nucleic acid molecule described in claim 8. [Claim 10] The nucleic acid delivery carrier according to claim 9, wherein the nucleic acid delivery carrier includes one derived from an adenovirus, an adeno-associated virus, a lentivirus, or another acceptable nucleic acid delivery carrier. [Claim 11] The bispecific binding molecule according to claim 1, wherein the dimer is a homodimer or a heterodimer. [Claim 12] A host cell containing an isolated nucleic acid molecule as described in claim 8. [Claim 13] A drug composition comprising the bispecific binding molecule described in claim 1 and a pharmaceutically acceptable carrier. [Claim 14] The drug composition according to claim 13, characterized in that it is in the form of a tablet, powder, granules, pill, injection, suspension, powder, emulsion, aerosol, gel, eye drops, sustained-release agent, or sustained-release implant. [Claim 15] A kit comprising the drug composition according to claim 13, which is sealed in a container, The container is a glass ampoule, glass bottle, plastic ampoule, plastic bottle, plastic bag, or pre-filled syringe. [Claim 16] The bispecific binding molecule according to claim 1, for use in the manufacture of a pharmaceutical composition for treating or preventing diseases related to VEGFA or VEGFC. [Claim 17] The bispecific binding molecule according to claim 16, wherein the disease is selected from neovascular eye diseases and cancer-related indications. [Claim 18] The bispecific binding molecule according to claim 17, characterized in that the disease is selected from age-related macular degeneration, diabetic macular edema, diabetic retinopathy, neovascular glaucoma, retinal vein occlusion, corneal neovascularization, neovascularization after corneal transplantation, breast cancer, kidney cancer, ovarian cancer, fallopian tube cancer, peritoneal cancer, gastric cancer, colorectal cancer, non-small cell lung cancer, bladder cancer, pancreatic cancer, liver cancer, cervical cancer, and glioblastoma.

Images