RNA-induced translational silencing and cellular apoptosis

• US 8,173,616 B2
• Filed: 05/01/2009
• Issued: 05/08/2012
• Est. Priority Date: 05/02/2008
• Status: Active Grant

• Current Assignee: Brigham and Women's Hospital Incorporated
• Original Assignee: Brigham and Women's Hospital Incorporated
• Inventors: Anderson, Paul, Yamasaki, Satoshi
• Primary Examiner(s): Gibbs, Terra Cotta
• Application Number:
  US12/990,493
• Publication Number:
  US 20110046209A1
• Time in Patent Office: 1,103 Days
• Field of Search: 514/44, 536/24.5
• US Class Current: 514/44.A
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 31/20 for DNA viruses
• CPC Class Codes: A61P 31/22 for herpes viruses
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• 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 2310/11 Antisense
• CPC Class Codes: C12N 2320/30 Special therapeutic applications

Abstract

The invention is directed to RNA molecules that can be used to inhibit protein synthesis and to induce cells to undergo apoptosis. It also includes pharmaceutical compositions containing the RNAs that can be used in treating or preventing tumors; abnormal dermatological growths and viral infections.

Claims

1. A method of inducing apoptosis or inhibiting protein synthesis in a population of cells, comprising administering to said cells an effective amount of an RNA molecule 18-35 nucleotides in length, wherein said RNA molecule has a sequence that is at least 90% identical to a sequence found within the first 40 nucleotides at the 5′

end of a human stress-induced transfer RNA (tiRNA).

2. The method of claim 1, wherein said RNA is selected from the group consisting of:

SEQ ID NO;

1-SEQ ID NO;

132.

3. The method of claim 2, wherein said RNA is present as part of a solution, suspension or emulsion at a concentration of 10 ng/ml-10 μ

g/ml.

4. The method of claim 2, wherein said cells are contacted with 1 μ

g-500 μ

g of RNA per million cells.

5. The method of claim 2, wherein said population of cells are in vitro.

6. The method of claim 2, wherein said population of cells are part of a benign or malignant tumor, an abnormal growth on the skin of a subject or are cells that are otherwise growing in vivo in an abnormally excessive manner.

17. The method of claim 2, wherein said RNA molecule has a sequence consisting of the sequence of any one of SEQ ID NO:

17-SEQ ID NO;

132.

18. The method of claim 2, wherein said RNA molecule has a sequence consisting of the sequence of any one of SEQ ID NO:

1-SEQ ID NO;

16.

7. A therapeutic method comprising administering to a patient an effective amount of a pharmaceutical composition comprising an RNA molecule 18-35 nucleotides in length, wherein said RNA molecule has a sequence that is at least 90% identical to a sequence found within the first 40 nucleotides at the 5′

end of a human stress-induced transfer RNA (tiRNA).

8. The therapeutic method of claim 7, wherein said pharmaceutical composition is in the form of a solution, cream, lotion, suspension, emulsion or gel, and said patient is treated for an abnormal dermatological growth by topically applying said composition.

9. The therapeutic method of claim 8, wherein said abnormal dermatological growth is a melanoma and wherein said patient is administered an RNA selected from the group consisting of:

SEQ ID NO;

1-SEQ ID NO;

132.

10. The therapeutic method of claim 7, wherein said pharmaceutical composition is in the form of an injectable composition and said patient is treated for a benign or malignant tumor by injecting said tumor with said composition.

11. The therapeutic method of claim 10, wherein said benign or malignant tumor is injected with 0.1-100 ml of said pharmaceutical composition per gram of tumor and wherein said patient is administered an RNA selected from the group consisting of:

SEQ ID NO;

1-SEQ ID NO;

132.

12. The therapeutic method of claim 7, wherein said therapeutic method comprises treating viral infection in a population of eukaryotic cells, by administering said pharmaceutical composition to said cells.

13. The therapeutic method of claim 12, wherein said viral infection is caused by a sexually transmitted virus or a virus that infects the eye of patients and wherein said RNA is selected from the group consisting of:

SEQ ID NO;

1-SEQ ID NO;

132.

14. The therapeutic method of claim 12, wherein said viral infection is caused by a virus selected from the group consisting of:

herpes simplex virus type 1;

herpes simplex virus type 2;

human papillomavirus;

human immunodeficiency virus; and

human cytomegalovirus and wherein said RNA is selected from the group consisting of;

SEQ ID NO;

1-SEQ ID NO;

132.

15. The therapeutic method of claim 12, wherein said RNA is part of a solution, suspension, emulsion, cream, lotion, ointment, or gel and is present at a concentration of 1 ng/ml-100 μ

g/ml.

16. The therapeutic method of claim 12, wherein said cells are contacted with 1 μ

g-500 μ

g of RNA per million cells.

19. The therapeutic method of claim 8, wherein said RNA molecule has a sequence consisting of the sequence of any one of SEQ ID NO:

17-SEQ ID NO;

132.

20. The therapeutic method of claim 8, wherein said RNA molecule has a sequence consisting of the sequence of any one of SEQ ID NO:

1-SEQ ID NO;

16.