Reduced size self-delivering RNAi compounds

• US 9,080,171 B2
• Filed: 03/24/2011
• Issued: 07/14/2015
• Est. Priority Date: 03/24/2010
• Status: Active Grant

• Current Assignee: Phio Pharmaceuticals Corp.
• Original Assignee: Phio Pharmaceuticals Corp.
• Inventors: Khvorova, Anastasia, Salomon, William, Kamens, Joanne, Samarsky, Dmitry, Woolf, Tod M., Cardia, James
• Primary Examiner(s): WHITEMAN, BRIAN A
• Application Number:
  US13/636,728
• Publication Number:
  US 20130131141A1
• Time in Patent Office: 1,573 Days
• US Class Current: 1/1
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07H 21/02 with ribosyl as saccharide radical
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/32 of the sugar
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 2'-R Modification
• CPC Class Codes: C12N 2310/334 Modified C
• CPC Class Codes: C12N 2310/335 Modified T or U
• CPC Class Codes: C12N 2310/344 Position-specific modifications, e.g. on every purine, at the 3'-end
• CPC Class Codes: C12N 2310/351 Conjugate
• CPC Class Codes: C12N 2310/3515 Lipophilic moiety, e.g. cholesterol
• CPC Class Codes: C12N 2310/3521 Methyl
• CPC Class Codes: C12N 2310/3533 Halogen
• CPC Class Codes: C12N 2320/32 Special delivery means, e.g. tissue-specific
• CPC Class Codes: C12N 2320/51 modulating the chemical stability, e.g. nuclease-resistance

Abstract

The present invention relates to methods for in vivo administration of sd-rxRNA molecules.

Claims

1. An sd-rxRNA² 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.