Patent EP-4316502-A2 (EP)
Title: Means and methods for the treatment of angiogenesis-, fibrosis- and cancer-related diseases with protein oligomers comprising nc-1-fc
Inventor: JAVAHERIAN KASHI
IPC: A61K38/00
Filed: 2019-04-16
Abstract: The present invention pertains to a method for producing a protein oligomer comprising at least two, and preferably three heterodimeric human NC-1-Fc proteins, the method comprising: a) culturing a host cell expressing (i) a fusion protein comprising, from N- to C-terminus, human NC-1 from collagen 18 fused to human IgG1 Fc with "knob" mutations, or human IgG1 Fc with "knob" mutations fused to human NC-1 from collagen 18, and (ii) human IgG1 Fc with "hole" mutations, under conditions which allow the formation of a protein oligomer comprising at least two, and preferably three heterodimeric human NC-1-Fc proteins, and wherein the fusion protein of (i) and the human IgG1 Fc with "hole" mutations of (ii) are expressed in a ratio of 2:1 or higher, and b) obtaining from the host cell of step a) the protein oligomer comprising at least two, and preferably three heterodimeric human NC-1-Fc proteins. The invention further relates to a method for producing a protein oligomer comprising at least two, and preferably three monomeric human NC-1-Fc proteins, the method comprising: a) culturing a host cell expressing a fusion protein comprising, from N-to C-terminus, human NC-1 from collagen 18 fused to human IgG1 Fc, or human IgG1 Fc fused to human NC-1 from collagen 18, wherein the human IgG1 Fc comprises at least one monomeric mutation, preferably the monomeric mutation F405R, and one or more half-life extension mutations, preferably the half-life extension mutations M252Y, S254T and T256E, under conditions which allow the formation of a protein oligomer comprising at least two, and preferably three monomeric human NC-1-Fc proteins, and b) obtaining from the host cell of step a) the protein oligomer comprising at least two, and preferably three monomeric human NC-1-Fc proteins. Furthermore, the invention provides for a protein oligomer comprising at least two, and preferably three heterodimeric human NC-1-Fc proteins for use in treating, ameliorating or preventing an angiogenesis-related disease, wherein the heterodimeric human NC-1-Fc protein comprises (i) a fusion protein comprising, from N- to C-terminus, human NC-1 from collagen 18 fused to human IgG1 Fc with "knob" mutations, or human IgG1 Fc with "knob" mutations fused to human NC-1 from collagen 18, and (ii) the human IgG1 Fc with "hole" mutations, wherein the "knob" mutations and "hole" mutations in human IgG1 Fc are defined as set forth in the present application. The invention also concerns a monomeric NC-1-Fc protein, comprising human NC-1 from collagen 18 and a Fc domain from human IgG1, wherein the Fc domain from human IgG1 comprises at least one monomeric mutation, preferably the monomeric mutation F405R, and one or more half-life extension mutations, preferably the half-life extension mutations M252Y, S254T and T256E. The invention also relates to a protein oligomer comprising at least two of said monomeric human NC-1-Fc fusion proteins for use as a medicament, preferably for treating an angiogenesis-related disease, fibrosis or a fibrosis-associated disease, vascular endothelial growth factor (VEGF)-related disease or matrix metalloproteinase (MMP)-related disease.