Reduced size self-delivering RNAi compounds
The present invention relates to methods for in vivo administration of sd-rxRNA molecules.
1. An sd-rxRNA 2 comprising
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 isolated double stranded nucleic acid molecule are modified, wherein at least two Us and/or Cs include a hydrophobic modification selected from the group consisting of a thiophene, ethynyl, isobutyl benzyl and imidazole modification, and wherein the hydrophobic modifications are located on positions 4 or 5 of the Us and/or Cs.
2. The sd-rxRNA® of claim 1 , wherein at least 60% of the nucleotides are modified.
3. The sd-rxRNA® of claim 1 , wherein the modifications include at least one 2′F or 2′O methyl modifications.
4. The sd-rxRNA® of claim 1 , wherein a plurality of U's and/or C's include a hydrophobic modification, optionally selected from the group consisting of a thiophene, ethynyl, isobutyl benzyl, and imidazole modification, and wherein the hydrophobic modifications are located on positions 4 or 5 of U's and/or C's.
5. The sd-rxRNA® of claim 1 , wherein the sd-rxRNA is attached to a linker.
6. The sd-rxRNA® of claim 5 , wherein the sd-rxRNA is linked to cholesterol, and/or wherein the guide strand contains a single stranded region which contains at least 5 phosphorothioate modifications.
7. The sd-rxRNA® of claim 5 , wherein the linker is protonatable.
8. The sd-rxRNA® of claim 1 , wherein the sd-rxRNA® is formulated for intravenous administration, subcutaneous administration or intrathecal administration and/or wherein the half-life of the sd-rxRNA® in serum is longer than 12 hours.
9. The sd-rxRNA® of claim 1 , wherein the guide strand is 19, 20, 21, 22 or 23 nucleotides long, the passenger strand is 13 nucleotides long, and at least 50% of the nucleotides within the double stranded nucleic acid molecule are phosphorothioate modified.
10. The sd-rxRNA® of claim 1 , wherein at least 50% of the nucleotides within the sd-rxRNA® are phosphorothioate modified.
11. A composition comprising the sd-rxRNA® of claim 1 .
12. The composition of claim 11 , wherein the composition comprises two or more different sd-rxRNA® molecules.
13. A method for inhibiting the expression of a target gene of a mammal, comprising:
administering to a mammal an isolated double stranded nucleic acid molecule according to claim claim 9 .
14. The method of claim 13 , wherein the isolated double stranded nucleic acid molecule is delivered to the brain or spinal cord of the mammal.
15. The method of claim 14 , wherein the isolated double stranded nucleic acid molecule is directed against a target gene encoding SOD1.