Age-related macular degeneration treatment

Abstract

This invention is directed to an RNA interference (RNAi) agent and the use of that RNAi agent to treat Age-related Macular Degeneration, as well as pharmaceutical compositions containing the RNAi agents of the invention. The RNAi agent is a DNA-directed RNA interference (ddRNAi) agent (being an RNA molecule), together with an expression cassette or construct to express that agent in a cell (including in vivo), for inhibiting, preventing or reducing expression of an AMD associated gene. Preferably that AMD associated gene is one that is associated with wet AMD.

1 . A DNA-directed RNA interference (ddRNAi) agent for inhibiting expression of one or more target sequences in an AMD associated gene, the ddRNAi agent comprising:
 a first effector sequence of at least 17 nucleotides in length; and   a first effector complement sequence;   
       wherein the effector sequence is substantially complementary to one or more target regions in a transcript of the one or more target sequences. 
     
     
         2 . A ddRNAi agent according to  claim 1 , comprising a second effector sequence and second effector complement sequence. 
     
     
         3 . A ddRNAi agent according to  claim 2  comprising, in a 5′ to 3′ direction:
 (a) a first effector sequence of at least 17 nucleotides in length;
 a second effector sequence of at least 17 nucleotides in length; 
 a second effector complement sequence; and 
 a first effector complement sequence; or 
 
 (b) a first effector sequence of at least 17 nucleotides in length;
 a first effector complement sequence; 
 a second effector sequence of at least 17 nucleotides in length; and 
 a second effector complement sequence; 
 
 wherein each effector sequence is substantially complementary to one or more target regions in a transcript of the one or more target sequences. 
 
     
     
         4 . (canceled) 
     
     
         5 . A ddRNAi agent according to  claim 1 , wherein:
 (i) the AMD associated gene is selected from the group consisting of VEGF-A, VEGFR2, PDGFR-β and CFB;   (ii) the target sequences are selected from the group consisting of any 10 or more contiguous nucleotides within a sequence set forth in any one of SEQ ID NOS: 1-39;   (iii) the AMD associated gene is VEGF-A and each effector sequence is selected from the sequences set forth in SEQ ID NOS: 40-49;   (iv) the AMD associated gene is VEGFR2, and each effector sequence is selected from the sequences set forth in SEQ ID NOS: 50-59;   (v) the AMD associated gene is PDGFR-β and each effector sequence is selected from the sequences set forth in SEQ ID NOS: 60-69; or   (vi) the AMD associated gene is CFB and each effector sequence is selected from the sequences set forth in SEQ ID NOS: 70-78.   
     
     
         6 - 10 . (canceled) 
     
     
         11 . A ddRNAi agent according to  claim 1 , wherein the agent is expressed within a miRNA structure. 
     
     
         12 . A ddRNAi agent according to  claim 1 , wherein the AMD is wet AMD. 
     
     
         13 . A ddRNAi expression cassette for expressing a ddRNAi agent according to  claim 1 , the expression cassette comprising (in no particular order):
 one or more promoter sequences;   one or more DNA sequences that encode for one or more effector sequences; and   one or more DNA sequences that encode for one or more effector complement sequences;   and optionally:   one or more terminator sequences;   one or more DNA sequences that encode for loop sequences, spacer sequences, or both;   one or more enhancer sequences; and/or   miRNA encoding (ME) sequences.   
     
     
         14 . (canceled) 
     
     
         15 . A ddRNAi expression construct comprising a ddRNAi expression cassette according to  claim 13 . 
     
     
         16 . A ddRNAi expression construct according to  claim 15 , wherein the construct is a viral delivery construct. 
     
     
         17 . A method of treating AMD in a subject comprising administering a therapeutically effective amount of a ddRNAi expression construct of  claim 15 , optionally wherein the AMD is wet AMD. 
     
     
         18 . (canceled) 
     
     
         19 . A method of treating choroidal neovascularisation in a subject comprising administering a therapeutically effective amount of a ddRNAi expression construct of  claim 15 . 
     
     
         20 . A method of reducing drusen deposits in a subject comprising administering a therapeutically effective amount of a ddRNAi expression construct of  claim 15 . 
     
     
         21 . A method according to  claim 17 , wherein the ddRNAi expression construct is administered to the subject's eye/s by intravitreal injection. 
     
     
         22 . A pharmaceutical composition comprising a ddRNAi expression construct of  claim 15 , and a pharmaceutically acceptable carrier or diluent. 
     
     
         23 . A ddRNAi agent according to  claim 2 , wherein:
 (i) the AMD associated gene is selected from the group consisting of VEGF-A, VEGFR2, PDGFR-β and CFB;   (ii) the target sequences are selected from the group consisting of any 10 or more contiguous nucleotides within a sequence set forth in any one of SEQ ID NOS: 1-39;   (iii) the AMD associated gene is VEGF-A and each effector sequence is selected from the sequences set forth in SEQ ID NOS: 40-49;   (iv) the AMD associated gene is VEGFR2, and each effector sequence is selected from the sequences set forth in SEQ ID NOS: 50-59;   (v) the AMD associated gene is PDGFR-β and each effector sequence is selected from the sequences set forth in SEQ ID NOS: 60-69; or   (vi) the AMD associated gene is CFB and each effector sequence is selected from the sequences set forth in SEQ ID NOS: 70-78.   
     
     
         24 . A ddRNAi expression cassette for expressing a ddRNAi agent according to  claim 2 , the expression cassette comprising (in no particular order):
 one or more promoter sequences;   one or more DNA sequences that encode for one or more effector sequences; and   one or more DNA sequences that encode for one or more effector complement sequences;   and optionally:   one or more terminator sequences;   one or more DNA sequences that encode for loop sequences, spacer sequences, or both;   one or more enhancer sequences; and/or   miRNA encoding (ME) sequences.   
     
     
         25 . A ddRNAi expression construct comprising a ddRNAi expression cassette according to  claim 24 , optionally comprising miRNA encoding (ME) sequences. 
     
     
         26 . A ddRNAi expression construct according to  claim 25 , wherein the construct is a viral delivery construct. 
     
     
         27 . A method of treating AMD in a subject comprising administering a therapeutically effective amount of a ddRNAi expression construct of  claim 25 . 
     
     
         28 . A method according to  claim 27  wherein the AMD is wet AMD. 
     
     
         29 . A method of treating choroidal neovascularisation in a subject comprising administering a therapeutically effective amount of a ddRNAi expression construct of  claim 25 . 
     
     
         30 . A method of reducing drusen deposits in a subject comprising administering a therapeutically effective amount of a ddRNAi expression construct of  claim 25 . 
     
     
         31 . A method according to  claim 27 , wherein the ddRNAi expression construct is administered to the subject's eye/s by intravitreal injection. 
     
     
         32 . A pharmaceutical composition comprising a ddRNAi expression construct of  claim 25 , and a pharmaceutically acceptable carrier or diluent.