Compositions and methods for treating and preventing macular degeneration

Abstract

Compositions and methods for treating macular degeneration are disclosed. The methods utilize gene delivery to human eyes of soluble Flt-1 receptors, as well fusion proteins including a soluble Flt-1 receptor.

1 . A method of treating macular degeneration in a human subject comprising administering to the diseased eye of the subject a composition comprising a recombinant adeno-associated virus (rAAV) virion comprising a polynucleotide encoding a soluble protein comprising at least one domain of VEGFR-1 (Flt-1) capable of modulating VEGF activity, wherein from about 1×10 7  to about 1×10 13 , or less than about 2×10 10  rAAV virions are delivered to the eye. 
     
     
         2 . The method of  claim 1 , wherein the method comprises reducing intraocular pressure, retinal thickness, subretinal fluids, or intraretinal fluids. 
     
     
         3 . A method of treating macular edema in a human subject comprising administering to the diseased eye of the subject a composition comprising a recombinant adeno-associated virus (rAAV) virion comprising a polynucleotide encoding a soluble protein comprising at least one domain of VEGFR-1 (Flt-1) capable of modulating VEGF activity, wherein from about 1×10 7  to about 1×10 13 , or less than about 2×10 10  rAAV virions are delivered to the eye. 
     
     
         4 . The method of  claim 3 , wherein the method comprises reducing intraocular pressure, retinal thickness, subretinal fluids, or intraretinal fluids. 
     
     
         5 - 10 . (canceled) 
     
     
         11 . The method of  claim 1 , wherein from about 2×10 8  to less than 2×10 10  rAAV virions are delivered to the eye. 
     
     
         12 . The method of  claim 1 , wherein up to about 2×10 8  rAAV virions are delivered to the eye. 
     
     
         13 . The method of  claim 1 , wherein up to about 2×10 9  rAAV virions are delivered to the eye. 
     
     
         14 . The method of  claim 1 , wherein the composition further comprises an opthalmalogically acceptable vehicle. 
     
     
         15 . The method of  claim 1 , wherein a single intravitreal injection of rAAV virions is administered to the eye. 
     
     
         16 . The method of  claim 1 , wherein the soluble protein comprises:
 (a) the at least one domain of Flt-1;   (b) a multimerization domain derived from an immunoglobulin heavy chain; and   (c) a linker 5-25 amino acid residues in length linking (a) to (b),   wherein when the soluble protein is expressed, a multimer of the soluble protein is produced.   
     
     
         17 . The method of  claim 16 , wherein the at least one domain comprises domain 2 of Flt-1. 
     
     
         18 . The method of  claim 16 , wherein the multimer is a homodimer. 
     
     
         19 . The method of  claim 16 , wherein the multimerization domain comprises the Fc region of an IgG, or an active fragment thereof. 
     
     
         20 . The method of  claim 16 , wherein the multimerization domain comprises the CH3 domain of an IgG, or an active fragment thereof. 
     
     
         21 . The method of  claim 16 , wherein the multimerization domain is from an IgG1, an IgG2, an IgG3 or an IgG4. 
     
     
         22 . The method of  claim 21 , wherein the multimerization domain is from the constant region of an IgG1 heavy chain. 
     
     
         23 . The method  claim 16  wherein the linker is selected from the group consisting of: 
       
         
           
                 
               
                   (SEQ ID NO: 1) 
                 
                   gly 9 ; 
                 
                     
                 
                   (SEQ ID NO: 2) 
                 
                   glu 9 ; 
                 
                     
                 
                   (SEQ ID NO: 3) 
                 
                   ser 9 ; 
                 
                     
                 
                   (SEQ ID NO: 4) 
                 
                   gly 5 cyspro 2 cys; 
                 
                     
                 
                   (SEQ ID NO: 5) 
                 
                   (gly 4 ser) 3 ; 
                 
                     
                 
                   (SEQ ID NO: 6) 
                 
                   SerCysValProLeuMetArgCysGlyGlyCysCysAsn; 
                 
                     
                 
                   (SEQ ID NO: 7) 
                 
                   ProSerCysValProLeuMetArgCysGlyGlyCysCysAsn; 
                 
                     
                 
                   (SEQ ID NO: 8) 
                 
                   GlyAspLeuIleTyrArgAsnGlnLys;  
                 
                   and 
                 
                     
                 
                   (SEQ ID NO: 9) 
                 
                   Gly 9 ProSerCysValProLeuMetArgCysGlyGlyCysCysAsn. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         24 . The method of  claim 16 , wherein the soluble protein has the formula X-Y-Z, wherein X comprises the IgG-like domain 2 of Flt-1, wherein Y is Gly 9 , and wherein Z is an IgG Fc region or an IgG CH3 region. 
     
     
         25 . The method of  claim 16 , wherein the multimerization domain is humanized. 
     
     
         26 . The method of  claim 16 , wherein the soluble protein comprises an amino acid sequence selected from the group consisting of (a) the amino acid sequence depicted in  FIGS. 2A-2B  (SEQ ID NO:11); (b) the amino acid sequence depicted in  FIG. 6  (SEQ ID NO:15); (c) the amino acid sequence depicted in  FIG. 8  (SEQ ID NO:17); (d) the amino acid sequence depicted in  FIG. 12  (SEQ ID NO:21); and (e) an active variant of (a), (b), (c) or (d) having at least 90% sequence identity thereto. 
     
     
         27 . The method of  claim 1 , wherein the macular degeneration is age-related macular degeneration (AMD). 
     
     
         28 . The method of  claim 27 , wherein the macular degeneration is wet AMD. 
     
     
         29 . The method of  claim 1 , wherein the rAAV virion is derived from an AAV serotype selected from AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAVrh8, AAVrh8R, AAV10, AAVrh10, AAV11 or AAV12. 
     
     
         30 . The method of  claim 29 , wherein the rAAV virion is derived from AAV2. 
     
     
         31 - 60 . (canceled) 
     
     
         61 . The method of  claim 3 , wherein from about 2×10 8  to less than 2×10 10  rAAV virions are delivered to the eye. 
     
     
         62 . The method of  claim 3 , wherein up to about 2×10 8  rAAV virions are delivered to the eye. 
     
     
         63 . The method of  claim 3 , wherein up to about 2×10 9  rAAV virions are delivered to the eye. 
     
     
         64 . The method of  claim 3 , wherein the composition further comprises an opthalmalogically acceptable vehicle. 
     
     
         65 . The method of  claim 3 , wherein a single intravitreal injection of rAAV virions is administered to the eye. 
     
     
         66 . The method of  claim 3 , wherein the soluble protein comprises:
 (a) the at least one domain of Flt-1;   (b) a multimerization domain derived from an immunoglobulin heavy chain; and   (c) a linker 5-25 amino acid residues in length linking (a) to (b),   wherein when the soluble protein is expressed, a multimer of the soluble protein is produced.   
     
     
         67 . The method of  claim 66 , wherein the at least one domain comprises domain 2 of Flt-1. 
     
     
         68 . The method of  claim 66 , wherein the multimer is a homodimer. 
     
     
         69 . The method of  claim 66 , wherein the multimerization domain comprises the Fc region of an IgG, or an active fragment thereof. 
     
     
         70 . The method of  claim 66 , wherein the multimerization domain comprises the CH3 domain of an IgG, or an active fragment thereof. 
     
     
         71 . The method of  claim 66 , wherein the multimerization domain is from an IgG1, an IgG2, an IgG3 or an IgG4. 
     
     
         72 . The method of  claim 71 , wherein the multimerization domain is from the constant region of an IgG1 heavy chain. 
     
     
         73 . The method of  claim 66  wherein the linker is selected from the group consisting of: 
       
         
           
                 
               
                   (SEQ ID NO: 1) 
                 
                   gly 9 ; 
                 
                     
                 
                   (SEQ ID NO: 2) 
                 
                   glu 9 ; 
                 
                     
                 
                   (SEQ ID NO: 3) 
                 
                   ser 9 ; 
                 
                     
                 
                   (SEQ ID NO: 4) 
                 
                   gly 5 cyspro 2 cys; 
                 
                     
                 
                   (SEQ ID NO: 5) 
                 
                   (gly 4 ser) 3 ; 
                 
                     
                 
                   (SEQ ID NO: 6) 
                 
                   SerCysValProLeuMetArgCysGlyGlyCysCysAsn; 
                 
                     
                 
                   (SEQ ID NO: 7) 
                 
                   ProSerCysValProLeuMetArgCysGlyGlyCysCysAsn; 
                 
                     
                 
                   (SEQ ID NO: 8) 
                 
                   GlyAspLeuIleTyrArgAsnGlnLys;  
                 
                   and 
                 
                     
                 
                   (SEQ ID NO: 9) 
                 
                   Gly 9 ProSerCysValProLeuMetArgCysGlyGlyCysCysAsn. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         74 . The method of  claim 66 , wherein the soluble protein has the formula X-Y-Z, wherein X comprises the IgG-like domain 2 of Flt-1, wherein Y is Gly 9 , and wherein Z is an IgG Fc region or an IgG CH3 region. 
     
     
         75 . The method of  claim 66 , wherein the multimerization domain is humanized. 
     
     
         76 . The method of  claim 66 , wherein the soluble protein comprises an amino acid sequence selected from the group consisting of (a) the amino acid sequence depicted in  FIGS. 2A-2B  (SEQ ID NO:11); (b) the amino acid sequence depicted in  FIG. 6  (SEQ ID NO:15); (c) the amino acid sequence depicted in  FIG. 8  (SEQ ID NO:17); (d) the amino acid sequence depicted in  FIG. 12  (SEQ ID NO:21); and (e) an active variant of (a), (b), (c) or (d) having at least 90% sequence identity thereto. 
     
     
         77 . The method of  claim 3 , wherein the rAAV virion is derived from an AAV serotype selected from AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAVrh8, AAVrh8R, AAV10, AAVrh10, AAV 11 or AAV12. 
     
     
         78 . The method of  claim 77 , wherein the rAAV virion is derived from AAV2.