Inhibition of cell proliferation using antisense oligonucleotides

• US 5,756,476 A
• Filed: 01/26/1994
• Issued: 05/26/1998
• Est. Priority Date: 01/14/1992
• Status: Expired due to Term

• Current Assignee: United States Department Of Health And Human Services
• Original Assignee: United States Department Of Health And Human Services
• Inventors: Speir, Edith H., Unger, Ellis F., Epstein, Stephen E.
• Primary Examiner(s): RORIES, CHARLES
• Application Number:
  US08/187,785
• Time in Patent Office: 1,581 Days
• Field of Search: 536/22.1, 536/24.5, 514/44, 435/375, 435/377, 435/6
• US Class Current: 514/44.00A
• 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: 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 15/1135 against oncogenes or tumor suppressor genes
• CPC Class Codes: C12N 2310/314 Phosphoramidates
• CPC Class Codes: C12N 2799/021 for the expression of a heterologous nucleic acid

Abstract

The present invention provides a method of inhibiting restenosis of a blood vessel in a mammal after mechanical treatment of the vessel to reduce a stenosis. The method includes contacting proliferating smooth muscle cells in the vessel with an antisense oligonucleotide directed against a cellular division cycle gene product. This gene product can be one of the following: c-myc, PCNA, and cyclin B₁. The antisense oligonucleotide is used in an amount effective to inhibit translation of the cellular division cycle gene product in the cell.

Claims

1. A method of inhibiting the growth of a human smooth muscle cell, comprising contacting said cell with a synthetic antisense oligonucleotide directed against a c-myc gene product in an amount effective to inhibit translation of said c-myc gene product in said cell.

2. The method of claim 1 wherein said c-myc gene product has an AUG translation initiation codon, and said synthetic antisense oligonucleotide is complementary to a portion of a 27 nucleotide sequence of said c-myc gene product extending from two codons upstream of the AUG translation initiation codon to six codons downstream of the AUG translation initiation codon.

3. The method of claim 2 wherein said synthetic antisense oligonucleotide is complementary to a region of said c-myc gene product that extends from said AUG translation initiation codon to four codons downstream of said AUG translation initiation codon.

4. The method of claim 3 wherein said synthetic antisense oligonucleotide is chemically modified with phosphoramidate linkages to block exonucleolytic degradation.

5. The method of claim 2 wherein said synthetic antisense oligonucleotide is complementary to a region of said c-myc gene product that extends from one codon upstream of said AUG translation initiation codon to four codons downstream of said AUG translation initiation codon.

6. The method of claim 5 wherein said synthetic antisense oligonucleotide is chemically modified with phosphoramidate linkages to block exonucleolytic degradation.

7. A method of inhibiting the migration of a human smooth muscle cell, comprising contacting said cell with a synthetic antisense oligonucleotide directed against a c-myc gene product in an amount effective to inhibit translation of said c-myc gene product in said cell.

8. The method of claim 7 wherein said c-myc gene product has an AUG translation initiation codon, and said synthetic antisense oligonucleotide is complementary to a portion of a 27 nucleotide sequence of said c-myc gene product extending from two codons upstream of the AUG translation initiation codon to six codons downstream of the AUG translation initiation codon.

9. The method of claim 8 wherein said synthetic antisense oligonucleotide is complementary to a region of said c-myc gene product that extends from said AUG translation initiation codon to four codons downstream of said AUG translation initiation codon.

10. The method of claim 9 wherein said synthetic antisense oligonucleotide is chemically modified with phosphoramidate linkages to block exonucleolytic degradation.

11. The method of claim 9 wherein said synthetic antisense oligonucleotide is complementary to a region of said c-myc gene product that extends from one codon upstream of said AUG translation initiation codon to four codons downstream of said AUG translation initiation codon.

12. The method of claim 11 herein said synthetic antisense oligonucleotide is chemically modified with phosphoramidate linkages to block exonucleolytic degradation.

13. A method of inhibiting restenosis of a blood vessel in a human after mechanical treatment of the vessel to reduce a stenosis, the method comprising contacting the vessel with a synthetic antisense oligonucleotide directed against a c-myc gene product in an amount effective to inhibit translation of said c-myc gene product in the cells of the vessel.

14. The method of claim 13 wherein said c-myc gene product has an AUG translation initiation codon, and said synthetic antisense oligonucleotide is complementary to a portion of a 27 nucleotide sequence of said c-myc gene product extending from two codons upstream of the AUG translation initiation codon to six codons downstream of the AUG translation initiation codon.

15. The method of claim 14 wherein said synthetic antisense oligonucleotide is complementary to a region of said c-myc gene product that extends from said AUG translation initiation codon to four codons downstream of said AUG translation initiation codon.

16. The method of claim 15 wherein said synthetic antisense oligonucleotide is chemically modified with phosphoramidate linkages to block exonucleolytic degradation.

17. The method of claim 16 wherein said mechanical treatment is cardiac angioplasty.

18. The method of claim 16 wherein said synthetic antisense oligonucleotide is complementary to a region of said c-myc gene product that extends from one codon upstream of said AUG translation initiation codon to four codons downstream of said AUG translation initiation codon.

19. The method of claim 18 wherein said synthetic antisense oligonucleotide is chemically modified with phosphorapidate linkages to block exonucleolytic degradation.