Age-related macular degeneration treatment
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 VEGF-A, the ddRNAi agent comprising:
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 one of the first or second effector sequences is substantially complementary to a region of corresponding length within a transcript of the sequence set forth in SEQ ID NO: 8.
2. A ddRNAi agent according to claim 1 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 one of the first or second effector sequences is substantially complementary to a region of corresponding length within a transcript of the sequence set forth in SEQ ID NO: 8.
3. A ddRNAi agent according to claim 1 , wherein the effector sequence which is substantially complementary to a region of corresponding length within a transcript of the sequence set forth in SEQ ID NO: 8 is selected from the group consisting of the sequences set forth in SEQ ID NOS: 40-49.
4. A ddRNAi agent according to claim 1 , wherein the agent is expressed within a miRNA structure.
5. 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.
6. A ddRNAi expression construct comprising a ddRNAi expression cassette according to claim 5 , optionally comprising miRNA encoding (ME) sequences.
7. A ddRNAi expression construct according to claim 6 , wherein the construct is a viral delivery construct.
8. A method of treating AMD in a subject comprising administering a therapeutically effective amount of a ddRNAi expression construct of claim 6 , optionally wherein the AMD is wet AMD.
9. A method of treating choroidal neovascularisation in a subject comprising administering a therapeutically effective amount of a ddRNAi expression construct of claim 6 .
10. A method of reducing drusen deposits in a subject comprising administering a therapeutically effective amount of a ddRNAi expression construct of claim 6 .
11. A method according to claim 8 , wherein the ddRNAi expression construct is administered to the subject's eye/s by intravitreal injection.
12. A pharmaceutical composition comprising a ddRNAi expression construct of claim 6 , and a pharmaceutically acceptable carrier or diluent.
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. A ddRNAi expression construct comprising a ddRNAi expression cassette according to claim 13 , optionally comprising miRNA encoding (ME) sequences.
15. A ddRNAi expression construct according to claim 14 , wherein the construct is a viral delivery construct.
16. A method of treating AMD in a subject comprising administering a therapeutically effective amount of a ddRNAi expression construct of claim 14 .
17. A method according to claim 16 wherein the AMD is wet AMD.
18. A method of treating choroidal neovascularisation in a subject comprising administering a therapeutically effective amount of a ddRNAi expression construct of claim 14 .
19. A method of reducing drusen deposits in a subject comprising administering a therapeutically effective amount of a ddRNAi expression construct of claim 14 .
20. A method according to claim 16 , wherein the ddRNAi expression construct is administered to the subject's eye/s by intravitreal injection.
21. A pharmaceutical composition comprising a ddRNAi expression construct of claim 14 , and a pharmaceutically acceptable carrier or diluent.
22. A ddRNAi agent according to claim 1 , wherein one of the first or second effector sequences comprises at least 17 contiguous nucleotides of the sequence set forth in SEQ ID NO: 47.
23. A ddRNAi agent according to claim 1 , wherein one of the first or second effector sequences comprises the sequence set forth in SEQ ID NO: 47.