Antisense modulation of human mdm2 expression

• US 6,184,212 B1
• Filed: 03/26/1999
• Issued: 02/06/2001
• Est. Priority Date: 03/26/1998
• Status: Expired due to Fees

• Current Assignee: ISIS Pharmaceuticals Incorporated
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Nero, Pamela, Miraglia, Loren J., Monia, Brett P., Graham, Mark J., Cowsert, Lex M.
• Primary Examiner(s): Elliott, George C.
• Assistant Examiner(s): EPPS -SMITH, JANET L
• Application Number:
  US09/280,805
• Time in Patent Office: 683 Days
• Field of Search: 514/44, 435/375, 435/325, 435/91.1, 435/6, 536/23.1, 536/24.5, 536/24.3
• 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: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. 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 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 2'-R Modification
• CPC Class Codes: C12N 2310/335 Modified T or U
• CPC Class Codes: C12N 2310/341 Gapmers, i.e. of the type ===---===
• CPC Class Codes: C12N 2310/346 having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/3521 Methyl
• CPC Class Codes: C12N 2310/3525 MOE, methoxyethoxy
• CPC Class Codes: Y02P 20/582 Recycling of unreacted starting or intermediate materials

Abstract

Compounds, compositions and methods are provided for inhibiting the expression of human mdm2. The compositions include antisense compounds targeted to nucleic acids encoding mdm2. Methods of using these oligonucleotides for inhibition of mdm2 expression and for treatment of diseases such as cancers associated with overexpression of mdm2 are provided.

Claims

1. An antisense compound 8 to 30 nucleobases in length targeted to nucleobases 1-308 of the 5′

untranslated region, 1776-1806 of the translation termination codon region or 1818-2370 of the 3′

untranslated region of a nucleic acid molecule encoding human mdm2, wherein said antisense compound modulates the expression of human mdm2.

2. The antisense compound of claim 1 wherein said antisense compound inhibits the expression of human mdm2.

3. The antisense compound of claim 2 comprising SEQ ID NO:

4.

4. A method of reducing hyperproliferation of human cells comprising contacting proliferating human cells in vitro with the antisense compound of claim 2 or a composition comprising said antisense compound.

5. The antisense compound of claim 1 which is an antisense oligonucleotide.

6. The antisense compound of claim 1 which contains at least one phosphorothioate intersugar linkage.

7. The antisense compound of claim 1 which has at least one 2′

-O-methoxyethyl modification.

8. The antisense compound of claim 1 which contains at least one 5-methyl cytidine.

9. The antisense compound of claim 7 in which every 2′

-O-methoxyethyl modified residue is a 5-methyl cytidine.

10. A method of modulating the expression of human mdm2 in human cells or tissues comprising contacting said cells or tissues in vitro with the antisense compound of claim 1.

11. An antisense compound up to 30 nucleobases in length comprising at least an 8-nucleobase portion of SEQ ID NO:

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

7, SEQ ID NO;

17, SEQ ID NO;

18, SEQ ID NO;

19, SEQ ID NO;

21, SEQ ID NO;

36, SEQ ID NO;

52, SEQ ID NO;

216, SEQ ID NO;

246, SEQ ID NO;

251, SEQ ID NO;

260, or SEQ ID NO;

264 which inhibits the expression of human mdm2.

12. An antisense compound up to 30 nucleobases in length targeted to a 5′

untranslated region of a nucleic acid molecule encoding a S-mdm2 transcript, wherein said antisense compound inhibits the expression of said S-mdm2 transcript and comprises at least an 8 nucleobase portion of SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5 or SEQ ID NO;

7.

13. An antisense compound up to 30 nucleobases in length comprising at least an 8-nucleobase portion of SEQ ID NO:

15, SEQ ID NO;

56, SEQ ID NO;

57, SEQ ID NO;

120, SEQ ID NO;

126, SEQ ID NO;

160, SEQ ID NO;

177, SEQ ID NO;

180, or SEQ ID NO;

195.

14. The antisense compound of claim 13 which contains at least one phosphorothioate intersugar linkage.

15. The antisense compound of claim 13 which has at least one 2′

-O-methoxyethyl modification.

16. The antisense compound of claim 13 which contains at least one 5-methyl cytidine.

17. The antisense compound of claim 15 in which every 2′

-O-methoxyethyl modified residue is a 5-methyl cytidine.

18. A method of modulating the expression of human mdm2 in cells or tissues comprising contacting said cells or tissues in vitro with the antisense compound of claim 13.

19. A method of reducing hyperproliferation of human cells comprising contacting proliferating human cells in vitro with the antisense compound of claim 13.

20. An antisense compound consisting of SEQ ID NO:

60, 61, 69 or 70.