Reduced size self-delivering RNAi compounds

• US 10,240,149 B2
• Filed: 06/02/2015
• Issued: 03/26/2019
• 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
• Application Number:
  US14/729,006
• Publication Number:
  US 20160130578A1
• Time in Patent Office: 1,393 Days
• Field of Search: None
• 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. 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.

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.

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.

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.