Reduced size self-delivering RNAi compounds
The present invention relates to methods for in vivo administration of sd-rxRNA molecules.
1. A method for delivering a nucleic acid to a remote target tissue in a subject in need thereof, comprising
systemically administering to the subject an sd-rxRNA® in an effective amount to promote RNA interference by the sd-rxRNA® in the remote target tissue, wherein the sd-rxRNA® comprises a guide strand and a passenger strand, wherein the sd-rxRNA® includes a double-stranded region and a single stranded region wherein the double stranded region is from 8-15 nucleotides long, wherein the single stranded region is at the 3′ end of the guide strand and is 4-12 nucleotides long, wherein the single stranded region contains 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 phosphorothioate modifications, wherein at least 40% of the nucleotides of the sd-rxRNA® are modified, and wherein at least two Us and/or Cs include a hydrophobic modification selected from the group consisting of a thiophene, octyn-1-yl, and ethynyl modification, located on position 5 of the Us and/or Cs.
2. The method of claim 1 , wherein at least 60% of the nucleotides on the guide strand of the sd-rxRNA® are modified.
3. The method of claim 1 , wherein the modifications include at least one 2′F or 2′O methyl modification.
4. The method of claim 1 , wherein a plurality of Us and/or Cs include a hydrophobic modification, and wherein each of the hydrophobic modifications is selected from the group consisting of a thiophene, octyn-1-yl, and ethynyl modification.
5. The method of claim 1 , wherein the sd-rxRNA® is attached to a linker.
6. The method of claim 1 , wherein the sd-rxRNA® is linked at the 3′ end to cholesterol, and/or wherein the guide strand contains at least 5 phosphorothioate modifications.
7. The method of claim 1 , wherein the guide strand of the sd-rxRNA® contains at least two single stranded regions.
8. The method of claim 1 , wherein the sd-rxRNA® is delivered to the liver, heart, brain, lung or fat.
9. A method comprising:
administering to a subject having a tumor an sd-rxRNA® in an effective amount to promote RNA interference by the sd-rxRNA® in the tumor, wherein the sd-rxRNA® comprises a guide strand and a passenger strand, wherein the sd-rxRNA® includes a double-stranded region and a single stranded region wherein the double stranded region is from 8-15 nucleotides long, wherein the single stranded region is at the 3′ end of the guide strand and is 4-12 nucleotides long, wherein the single stranded region contains 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 phosphorothioate modifications, and wherein at least 40% of the nucleotides of the sd-rxRNA® are modified, and wherein at least two Us and/or Cs include a hydrophobic modification selected from the group consisting of a thiophene, octyn-1-yl, and ethynyl modification, located on position 5 of the Us and/or Cs.
10. The method of claim 9 , wherein the sd-rxRNA® is administered to the tumor through intravenous administration.
11. The method of claim 9 , wherein the sd-rxRNA® is administered to the tumor through direct injection into the tumor.
12. The method of claim 9 , wherein the sd-rxRNA® is directed against a gene encoding for a protein selected from the group consisting of: VEGF/VEGFR, HER2, PDGF/PDGFR, HDAC, MET, c-kit, CDK, FLT-1, IGF/IGFR, FGF/FGFR, Ras/Raf, Abl, Bcl-2, Src, mTOR, PKC, MAPK, BIRC5, FAS, HIF1A, CDH16, MYC, HRAS and CTNNB1, or a combination thereof.
13. A method comprising:
administering an sd-rxRNA® to the central nervous system of a subject in need thereof, wherein the sd-rxRNA® comprises a guide strand and a passenger strand, wherein the sd-rxRNA® includes a double-stranded region and a single stranded region wherein the double stranded region is from 8-15 nucleotides long, wherein the single stranded region is at the 3′ end of the guide strand and is 4-12 nucleotides long, wherein the single stranded region contains 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 phosphorothioate modifications, and wherein at least 40% of the nucleotides of the sd-rxRNA® are modified, and wherein at least two Us and/or Cs include a hydrophobic modification selected from the group consisting of a thiophene, octyn-1-yl, and ethynyl modification, located on position 5 of the Us and/or Cs.
14. The method of claim 13 , wherein the sd-rxRNA® is administered by intrathecal delivery.
15. The method of claim 9 , wherein the modifications include at least one 2′F or 2′O methyl modification.
16. The method of claim 9 , wherein a plurality of Us and/or Cs include a hydrophobic modification, wherein each of the hydrophobic modifications is selected from the group consisting of a thiophene, octyn-1-yl, and ethynyl modification.
17. The method of claim 9 , wherein the sd-rxRNA® is linked at the 3′ end to cholesterol, and/or wherein the guide strand contains at least 5 phosphorothioate modifications.
18. The method of claim 13 , wherein the modifications include at least one 2′F or 2′O methyl modification.
19. The method of claim 13 , wherein a plurality of Us and/or Cs include a hydrophobic modification, wherein each of the hydrophobic modifications is selected from the group consisting of a thiophene, octyn-1-yl, and ethynyl modification.
20. The method of claim 13 , wherein the sd-rxRNA® is linked at the 3′ end to cholesterol, and/or wherein the guide strand contains at least 5 phosphorothioate modifications.
21. A method for delivering a nucleic acid to a remote target tissue in a subject in need thereof, comprising
systemically administering to the subject an sd-rxRNA® in an effective amount to promote RNA interference by the sd-rxRNA® in the remote target tissue, wherein the sd-rxRNA® comprises a guide strand and a passenger strand, wherein the sd-rxRNA® includes a double-stranded region and a single stranded region wherein the double stranded region is from 8-15 nucleotides long, wherein the single stranded region is at the 3′ end of the guide strand and is 4-12 nucleotides long, wherein the single stranded region contains 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 phosphorothioate modifications, wherein at least 40% of the nucleotides of the sd-rxRNA® are modified, wherein at least two Us include an imidazole modification, and wherein the imidazole modifications are located on position 5 of the Us.