EFFECTS OF APOLIPOPROTEIN B INHIBITION ON GENE EXPRESSION PROFILES IN ANIMALS

• US 20100331390A1
• Filed: 03/09/2010
• Published: 12/30/2010
• Est. Priority Date: 11/13/2002
• Status: Abandoned Application

• Current Assignee: Kastle Therapeutics, LLC.
• Original Assignee: Genzyme Corporation (Sanofi )
• Inventors: Graham, Mark J., Crooke, Rosanne M.
• Application Number:
  US12/720,581
• Publication Number:
  US 20100331390A1
• Time in Patent Office: Days
• 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 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• 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 2310/13 Decoys
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3341 5-Methylcytosine
• 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 2310/3525 MOE, methoxyethoxy
• CPC Class Codes: Y02P 20/582 Recycling of unreacted starting or intermediate materials

Abstract

Methods are provided for modulating the expression of genes involved in lipid metabolism, useful in the treatment of conditions associated with cardiovascular risk. Antisense oligonucleotides targeted to apolipoprotein B reduce the level of apolipoprotein B mRNA, lower serum cholesterol and shift liver gene expression profiles from those of an obese animal towards those of a lean animal. Further provided are methods for improving the cardiovascular risk of a subject through antisense inhibition of apolipoprotein B. Also provided are methods for employing antisense oligonucleotides targeted to apolipoprotein B to modulate a cellular pathway or metabolic process.

Claims

1. A method comprising contacting an animal with an antisense oligonucleotide 15-30 nucleobases in length, and modulating the level of a target gene mRNA,

wherein said antisense oligonucleotide reduces the level of apolipoprotein B mRNA and wherein said target gene is selected from the group consisting of Lcat, Lip1, Lipc, Ppara, Pparg, Pcx, Apoa4, Apoc1, Apoc2, Apoc4, Mttp, Prkaa1, Prkaa2, Prkab1, Prkag1, Srebp-1, Scd2, Scd1, Acad1, Acadm, Acads, Acox1, Cpt1a, Cpt2, Crat, Elovl2, Elovl3, Acadsb, Fads2, Fasn, Facl2, Facl4, Abcd2, Dbi, Fabp1, Fabp2, Fabp7, Acat-1, Acca-1, Cyp7a1, Cyp7b1, Soat2, Ldlr, Hmgcs1, Hmgcs2, Car5a, Gck, Gck and G6 pc.

2. The method of claim 1 which results in a shift a gene expression profile of an obese animal to that of a lean animal.

3. The method of claim 1 wherein the target gene mRNA is reduced in a time dependent manner.

4. The method of claim 3 wherein the target gene mRNA is reduced in a dose dependent manner.

5. The method of claim 1 wherein said antisense oligonucleotide comprises a chimeric oligonucleotide.

6. The method of claim 1 wherein said antisense oligonucleotide has at least one modified internucleoside linkage, sugar moiety or nucleobase.

7. The method of claim 1 wherein said antisense oligonucleotide has at least one 2′

-O-methoxyethyl sugar moiety.

8. The method of claim 1 wherein said antisense oligonucleotide has at least one phosphorothioate internucleoside linkage.

9. The method of claim 1 wherein at least one cytosine in said antisense oligonucleotide is a 5-methyl cytosine.

10. An antisense oligonucleotide 15-30 nucleobases in length targeted to a nucleic acid encoding apolipoprotein B that shifts a liver gene expression profile of an obese animal to that of a lean animal.

11. A method of lowering the cardiovascular risk profile of an individual, said individual having a high cardiovascular risk profile as defined by ATP III, comprising administering to said individual the compound of claim 10.

12. A method of altering a cellular pathway or metabolic process comprising contacting a cell with an antisense oligonucleotide that specifically hybridizes to and inhibits the expression of a nucleic acid molecule encoding apolipoprotein B, wherein the cellular pathway or metabolic process is apoptosis, angiogenesis, leptic secretion or T-cell co-stimulation.

13. The method of claim 12, wherein the antisense oligonucleotide comprises SEQ ID NO:

20.

14. The method of claim 12, wherein apoptosis is induced in said cells.

15. The method of claim 14 wherein said cells are cancer cells.

16. The method of claim 15 wherein said cancer cells are breast cancer cells.

17. The method of claim 12 wherein angiogenesis is inhibited.

18. The method of claim 12 wherein leptin secretion is increased.

19. The method of claim 12 wherein T-cell co-stimulation is inhibited.