Vlp for the treatment of a lysosomal storage disease

Abstract

The invention relates to virus like particles (VLP) associated with a lysosomal enzyme or an expression vector encoding a lysosomal enzyme which are used in a method for the treatment of a lysosomal storage disease. The invention also relates to a pharmaceutical composition for use in a method for the treatment of a lysosomal storage disease, to an expression vector encoding a lysosomal enzyme and to a method of associating a VLP with an expression vector encoding a lysosomal enzyme.

1 . A method of treating Metachromatic Leukodystrophy Disease (MLD), comprising administering a virus-like particle (VLP) to a subject in need thereof,
 wherein said VLP incorporates a lysosomal enzyme, or incorporates an expression vector comprising a nucleotide sequence encoding a lysosomal enzyme, and   wherein the VLP is derived from JCV.   
     
     
         2 . The method according to  claim 1 , wherein the VLP does not comprise viral genetic material, and the expression vector does not encode viral proteins. 
     
     
         3 . The method according to  claim 1 , wherein the subject is an animal. 
     
     
         4 . The method according to  claim 1 , wherein the subject has a neurologic deficit. 
     
     
         5 . The method according to  claim 1 , wherein the lysosomal enzyme is a human enzyme. 
     
     
         6 . The method according to  claim 1 , wherein the lysosomal enzyme is arylsulfatase A. 
     
     
         7 . The method according to  claim 1 , wherein the lysosomal enzyme comprises an amino acid sequence which is at least 80% identical to the amino acid sequence of SED NO: 1 over its entire length. 
     
     
         8 . The method according to  claim 1 , wherein the VLP incorporates an expression vector comprising a nucleotide sequence encoding a lysosomal enzyme, and wherein the expression vector has a size of less than 7 kb. 
     
     
         9 . The method according to  claim 1 , wherein the VLP incorporates an expression vector comprising a nucleotide sequence encoding a lysosomal enzyme, and wherein the expression vector has a promoter selected from the group consisting of a CMV and a CAG promoter. 
     
     
         10 . The method according to  claim 1 , wherein the VLP incorporates an expression vector comprising a nucleotide sequence encoding a lysosomal enzyme, and wherein the nucleotide sequence encoding the lysosomal enzyme is at least 70% identical to the nucleotide sequence of SEQ ID NO: 2 over its entire length. 
     
     
         11 . The method according to  claim 1 , wherein after said administration, the VLP crosses the blood-brain barrier to enter the CNS together with the lysosomal enzyme or the expression vector. 
     
     
         12 . The method according to  claim 11 , wherein the lysosomal enzyme or the expression vector enters at least one cell type selected from the group consisting of astrocytes, oligodendrocytes, microglia and neurons, after crossing said blood-brain barrier. 
     
     
         13 . The method according to  claim 1 , wherein the VLP is administered orally or parenterally. 
     
     
         14 . The method according to  claim 1 , wherein after said administration, a target cell is contacted with an effective amount of the lysosomal enzyme. 
     
     
         15 . The method according to  claim 1 , wherein the lysosomal enzyme has a therapeutically effective enzyme activity for at least 10 days. 
     
     
         16 . The method according to  claim 1 , wherein the VLP is composed of VP1 proteins of JC virus. 
     
     
         17 . The method according to  claim 16 , wherein the VP1 protein comprises an amino acid sequence which is at least 80% identical to the amino acid sequence of SEQ ID NO: 3 over its entire length. 
     
     
         18 . The method according to  claim 1 , wherein said VLP is administered in a pharmaceutical composition that further comprises at least one of a pharmaceutically acceptable carrier and an excipient. 
     
     
         19 . An expression vector comprising a nucleotide sequence encoding a lysosomal enzyme, and further comprising a promoter selected from the group consisting of CAG and CMV, wherein said expression vector has a size of less than 7 kb. 
     
     
         20 . The expression vector according to  claim 19 , wherein the nucleotide sequence encoding the lysosomal enzyme is at least 70% identical to the nucleotide sequence of SEQ ID NO: 2 over its entire length. 
     
     
         21 . A method of incorporating a lysosomal enzyme, or incorporating an expression vector comprising a nucleotide sequence encoding a lysosomal enzyme, into a VLP, wherein the method comprises:
 a) providing a composition comprising VP1 proteins,   b) exposing the VP1 proteins of the composition of a) to conditions inducing the VP1 to assemble into VLP,   c) exposing the VLP of the composition of b) to conditions disassembling the VLP into pentamers,   d) exposing the pentamers of the composition of c) to conditions inducing the pentamers to reassemble into VLP,   e) exposing the VLP of the composition of d) to conditions disassembling the VLP into pentamers, and   f) exposing the pentamers of the composition of e) to the lysosomal enzyme or the expression vector to conditions inducing the pentamers to assemble into a VLP associated with the lysosomal enzyme or the expression vector.   
     
     
         22 . A VLP produced by the method according to  claim 21 . 
     
     
         23 . A VLP incorporating a lysosomal enzyme, or incorporating an expression vector comprising a nucleotide sequence encoding a lysosomal enzyme, for use in the method of  claim 1 . 
     
     
         24 . The VLP according to  claim 23 , wherein the VLP does not comprise viral genetic material, and the expression vector does not encode viral proteins. 
     
     
         25 . The method according to  claim 1 , wherein said method does not comprise a step of increasing the permeability of a blood-brain barrier of the subject. 
     
     
         26 . The VLP according to  claim 23 , wherein the VLP is derived from JCV. 
     
     
         27 . The VLP according to  claim 23 , wherein the lysosomal enzyme is arylsulfatase A. 
     
     
         28 . The VLP according to  claim 23 , wherein the lysosomal enzyme comprises an amino acid sequence which is at least 80% identical to the amino acid sequence of SEQ ID NO: 1 over its entire length. 
     
     
         29 . The method according to  claim 14 , wherein the lysosomal enzyme is transiently expressed in the target cell. 
     
     
         30 . The VLP according to  claim 28 , wherein the lysosomal enzyme comprises an amino acid sequence which is at least 90% identical to the amino acid sequence of SEQ ID NO: 1 over its entire length. 
     
     
         31 - 32 . (canceled) 
     
     
         33 . The method according to  claim 1 , wherein the subject is a human. 
     
     
         34 . The method according to  claim 7 , wherein the lysosomal enzyme comprises an amino acid sequence which is at least 90% identical to the amino acid sequence of SEQ ID NO: 1 over its entire length. 
     
     
         35 . The method according to  claim 34 , wherein the lysosomal enzyme comprises the amino acid sequence of SEQ ID NO: 1. 
     
     
         36 . The method according to  claim 8 , wherein the expression vector has a size of less than 6 kb. 
     
     
         37 . The method according to  claim 10 , wherein the nucleotide sequence encoding the lysosomal enzyme is at least 80% identical to the nucleotide sequence of SEQ ID NO: 2 over its entire length. 
     
     
         38 . The method according to  claim 37 , wherein the nucleotide sequence encoding the lysosomal enzyme is at least 90% identical to the nucleotide sequence of SEQ ID NO: 2 over its entire length. 
     
     
         39 . The method according to  claim 38 , wherein the nucleotide sequence encoding the lysosomal enzyme comprises the nucleotide sequence of SEQ ID NO: 2. 
     
     
         40 . The method according to  claim 11 , wherein said blood-brain barrier is a physiologically intact blood-brain barrier. 
     
     
         41 . The method according to  claim 12 , wherein the lysosomal enzyme or the expression vector enters an oligodendrocyte after crossing said blood-brain barrier. 
     
     
         42 . The method according to  claim 1 , wherein the VLP is administered intravenously. 
     
     
         43 . The method according to  claim 15 , wherein the lysosomal enzyme has a therapeutically effective enzyme activity for at least 20 days. 
     
     
         44 . The method according to  claim 43 , wherein the lysosomal enzyme has a therapeutically effective enzyme activity for at least 30 days. 
     
     
         45 . The method according to  claim 17 , wherein the VP1 protein comprises an amino acid sequence which is at least 90% identical to the amino acid sequence according to SEQ ID NO: 3 over its entire length. 
     
     
         46 . The expression vector according to  claim 20 , wherein the nucleotide sequence encoding the lysosomal enzyme comprises a nucleotide sequence which is at least 80% identical to the nucleotide sequence of SEQ ID NO: 2 over its entire length. 
     
     
         47 . The expression vector according to  claim 46 , wherein the nucleotide sequence encoding the lysosomal enzyme comprises a nucleotide sequence which is at least 90% identical to the nucleotide sequence of SEQ ID NO: 2 over its entire length. 
     
     
         48 . The expression vector according to  claim 47 , wherein the nucleotide sequence encoding the lysosomal enzyme comprises the nucleotide sequence of SEQ ID NO: 2. 
     
     
         49 . The expression vector according to  claim 19 , wherein the lysosomal enzyme is human arylsulfatase A.