Method and reagent for inhibiting hepatitis C virus replication

• US 5,869,253 A
• Filed: 12/26/1996
• Issued: 02/09/1999
• Est. Priority Date: 05/14/1992
• Status: Expired due to Fees

• Current Assignee: Ribozyme Pharmaceuticals, Inc. (Alnylam Pharmaceuticals Incorporated)
• Original Assignee: Ribozyme Pharmaceuticals, Inc. (Alnylam Pharmaceuticals Incorporated)
• Inventors: Draper, Kenneth G.
• Primary Examiner(s): LeGuyader, John L.
• Application Number:
  US08/774,306
• Time in Patent Office: 775 Days
• Field of Search: 435/6, 435/91.31, 435/172.3, 435/320.1, 435/325, 435/366, 536/23.1, 536/24.5, 536/23.2, 514/44
• US Class Current: 435/5
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C12N 15/1131 against viruses
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2310/111 spanning the whole gene, or a large part of it
• CPC Class Codes: C12N 2310/12 catalytic nucleic acids, e.g. ribozymes
• CPC Class Codes: C12N 2310/121 Hammerhead
• CPC Class Codes: C12N 2310/122 Hairpin
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 2'-R Modification
• CPC Class Codes: C12N 2310/3521 Methyl
• CPC Class Codes: C12N 2310/3531 Hydrogen
• CPC Class Codes: C12N 2310/3533 Halogen
• CPC Class Codes: C12N 2310/3535 Nitrogen
• CPC Class Codes: C12N 2770/24222 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2770/24243 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12Q 1/701 : Specific hybridization probes
• CPC Class Codes: C12Q 1/707 : non-A, non-B Hepatitis, excluding hepatitis D

Abstract

An enzymatic RNA molecule which specifically cleaves RNA of a hepatitis C virus.

Claims

1. An enzymatic RNA molecule which specifically cleaves RNA encoding hepatitis C Virus (HCV) RNA, wherein said enzymatic RNA molecule comprises a substrate binding site and a nucleotide sequence within or surrounding said substrate binding site wherein said nucleotide sequence imparts to said enzymatic RNA molecule activity for the cleavage of said HCV RNA.

2. The enzymatic RNA molecule of claim 1, wherein said substrate binding site is complementary to said HCV RNA.

3. The enzymatic RNA molecule of claim 1, wherein said enzymatic RNA molecule is in a hammerhead motif.

4. The enzymatic RNA molecule of claim 1, wherein said enzymatic RNA molecule is in a hairpin motif.

5. The enzymatic RNA molecule of claim 1, wherein said substrate binding site comprises between 12 and 100 nucleotides complementary to said HCV RNA.

6. The enzymatic RNA molecule of claim 1, wherein said substrate binding site comprises between 14 and 24 nucleotides complementary to said HCV RNA.

7. An expression vector comprising nucleic acid sequence encoding one or more enzymatic RNA molecule of claim 1 in a manner which allows expression of said enzymatic RNA molecules.

8. The expression vector of claim 7, wherein said expression vector is a viral vector.

9. The expression vector of claim 8, wherein said viral vector is a retrovirus vector.

10. The enzymatic RNA molecule of claim 1, wherein said enzymatic RNA molecule is chemically synthesized.

11. The enzymatic RNA molecule of claim 1, wherein said enzymatic RNA molecule is in a purified form.

12. The enzymatic RNA molecule of claim 1, wherein said enzymatic RNA molecule is active in the presence of divalent metal ions.

13. The enzymatic RNA molecule of claim 12, wherein said divalent metal ion is magnesium.

14. The enzymatic RNA molecule of claim 1, wherein said enzymatic RNA molecule comprises a ribose modification.

15. The expression vector of claim 7, wherein said nucleic acid sequence encoding said enzymatic RNA molecule is under the control of a mammalian transcription promoter.

16. The expression vector of claim 15, wherein said mammalian transcription promoter is a cytomegalovirus promoter.

17. The enzymatic RNA molecule of claim 1, wherein said enzymatic RNA molecule comprises a 5'"'"'-cap.

18. The enzymatic RNA molecule of claim 1, wherein said enzymatic RNA molecule comprises a 3'"'"'-cap.

19. The enzymatic RNA molecule of claim 1, wherein said enzymatic RNA molecule comprises a 3'"'"'-polyA tail.

20. A method of cleaving HCV RNA comprising the step of contacting said HCV RNA with the enzymatic RNA molecule of claim 1 under conditions suitable for the cleavage of said HCV RNA.

21. The enzymatic RNA molecule of claim 1, wherein said enzymatic RNA molecule is capable of inhibiting replication of HCV.

22. The enzymatic RNA molecule of claim 21, wherein said enzymatic RNA molecule is in a hammerhead motif.