Multimeric constructs

Abstract

Multimeric fusion proteins of an Ig-like domain of Flt-1 are rendered functional by inclusion of a linker moiety. Vectors encoding the fusion proteins and host cells expressing the fusion proteins can be used therapeutically to block neovascularization in individuals with pathological conditions related to neovascularization. Such conditions include age-related macular degeneration, cancer, psoriasis, proliferative diabetic retinopathy, asthma, uveitis, osteoarthritis, and rheumatoid arthritis. The same means of multimerization used for an Ig like domain of Flt-1, i.e., a linker and a multimerization domain, can be used for other polypeptides, including extracellular receptors, antibody variable regions, cytokines, chemokines, and growth factors.

1 - 57 . (canceled) 
     
     
         58 . A method comprising: delivering a nucleic acid encoding a fusion protein of the formula X-Y-Z to a mammal, wherein X comprises a polypeptide that is an extracellular receptor, an antibody variable region, a cytokine, a chemokine, and a growth factor, wherein Y consists essentially of a 5-25 amino acid residue polypeptide, and wherein Z is an Fc portion, and whereby said fusion protein is expressed in the mammal. 
     
     
         59 . The method of  claim 58  wherein the fusion protein comprises a sequence selected from the group consisting of SEQ ID NO: 2, 8, 21, 23, and 25. 
     
     
         60 . The method of  claim 58 , wherein the mammal has wet age-related macular degeneration or proliferative diabetic retinopathy. 
     
     
         61 . The method of  claim 58 , wherein the mammal has cancer. 
     
     
         62 . The method of  claim 58 , wherein the mammal has rheumatoid arthritis. 
     
     
         63 . The method of  claim 58 , wherein the mammal has asthma. 
     
     
         64 . The method of  claim 58 , wherein the mammal has osteoarthritis. 
     
     
         65 - 71 . (canceled) 
     
     
         72 . The fusion protein of  claim 58 , wherein X comprises an extracellular receptor and said receptor is selected from the group consisting of a tyrosine kinase receptor and a serine threonine kinase receptor. 
     
     
         73 . The method of  claim 58 , wherein the extracellular receptor is a VEGF receptor. 
     
     
         74 . The method of  claim 58 , wherein X is the IgG-like domain 2 of VEGF-R1 (FLT-1). 
     
     
         75 . The method of  claim 58 , wherein the polypeptide Y is flexible. 
     
     
         76 . The method of  claim 58 , wherein the polypeptide Y is selected from the group consisting of gly9 (SEQ ID NO: 27), glu9 (SEQ ID NO: 28), ser9 (SEQ ID NO: 29), gly5cyspro2cys (SEQ ID NO: 30), (gly4ser)3 (SEQ ID NO: 31), SerCysValProLeuMetArgCysGlyGlyCysCysAsn (SEQ ID NO: 32), Pro Ser Cys Val Pro Leu Met Arg Cys Gly Gly Cys Cys Asn (SEQ ID NO: 13), Gly-Asp-Leu-Ile-Tyr-Arg-Asn-Gln-Lys (SEQ ID NO: 26), and Gly9ProSerCysValProLeuMetArgCysGlyGlyCysCysAsn (SEQ ID NO: 34). 
     
     
         77 . The method of  claim 58 , wherein the Fc is an IgG1 Fc. 
     
     
         78 . The method of  claim 58 , wherein Z is an IgG1 CH3 region. 
     
     
         79 . The method of  claim 58 , wherein Z is an IgG2 CH3 region. 
     
     
         80 . The method of  claim 58 , wherein the nucleic acid is in a vector. 
     
     
         81 . The method of  claim 80 , wherein the vector is a viral vector or a plasmid vector. 
     
     
         82 . The method of  claim 81 , wherein the viral vector is an adeno-associated virus (AAV) vector. 
     
     
         83 . The method of  claim 58 , wherein the nucleic is delivered by intravitreal injection to the mammal. 
     
     
         84 . The method of  claim 58 , wherein the mammal is a human.