Inhibition of PPAR gamma expression by specific osteogenic oxysterols

• US 8,022,052 B2
• Filed: 12/19/2007
• Issued: 09/20/2011
• Est. Priority Date: 12/19/2006
• Status: Active Grant

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Parhami, Farhad, Kim, Woo-Kyun, Nguyen, Khanhlinh, Jung, Michael E.
• Primary Examiner(s): HENLEY III, RAYMOND J
• Application Number:
  US12/520,281
• Publication Number:
  US 20100105645A1
• Time in Patent Office: 1,371 Days
• Field of Search: 514/170, 514/182
• US Class Current: 514/170
• CPC Class Codes: A61K 31/57 substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: C07J 1/00 Normal steroids containing carbon, hydrogen, halogen or oxygen, not substituted in position 17 beta by a carbon atom, e.g. estrane, androstane
• CPC Class Codes: G01N 2333/70567 Nuclear receptors, e.g. retinoic acid receptor [RAR], RXR, nuclear orphan receptors
• CPC Class Codes: G01N 2500/00 Screening for compounds of potential therapeutic value
• CPC Class Codes: G01N 2500/10 involving cells

Abstract

This invention relates to methods for using agents to inhibit peroxisome proliferator activated receptor expression.

Claims

1. A method for inhibiting expression of a peroxisome proliferator activated receptor (PPAR) in a cell, comprising:

contacting at least one oxysterol compound with the cell in an amount effective to inhibit expression of the PPAR; and

measuring inhibition of expression of the PPAR.

2. The method of claim 1, wherein the inhibited expression is of PPAR gamma.

3. The method of claim 1, wherein the cell is a marrow stromal cell (MSC) and the at least one oxysterol compound stimulates osteogenesis of the cell.

4. The method of claim 1, wherein the cell is a marrow stromal cell (MSC), and the at least one oxysterol compound has inhibits adipogenesis of the cell.

5. The method of claim 1, wherein the at least one oxysterol compound is an activator of the hedgehog signaling pathway.

6. The method of claim 1, wherein the at least one oxysterol compound is selected from the group consisting of 20(S)-hydroxycholesterol,

7. The method of claim 1, wherein the cell is a marrow stromal cell.

8. The method of claim 1, wherein contacting the at least one oxysterol compound with the cell inhibits expression and/or activity of a PPAR target gene;

and further comprising measuring inhibition of expression of the PPAR target gene.

9. The method of claim 8, wherein the PPAR target gene is a gene selected from the group consisting of adipocyte protein 2 (aP2) and lipoprotein lipase (LPL).

10. The method of claim 1, wherein the inhibition of PPAR expression is sufficient to inhibit a PPAR mediated response in the cell.

11. The method of claim 1, wherein the cell is in vitro.

12. The method of claim 1, wherein the cell is in a subject.

13. The method of claim 1, wherein the cell is in a human.

14. A method of treating a condition associated with increased adipogenesis in a subject, comprising administering to the subject an amount of an oxysterol compound effective to treat the condition, wherein the oxysterol compound has been shown to be in an amount that is effective to inhibit PPAR expression.

15. A method of treating a condition associated with excessive accumulation of intracellular and/or extracellular fats and/or lipids in a subject, comprising administering to the subject an amount of an oxysterol compound effective to treat the condition, wherein the oxysterol compound has been shown to be in an amount that is effective to inhibit PPAR expression.

16. A method for identifying an oxysterol compound that inhibits expression of PPAR, comprising:

screening a candidate oxysterol compound for the ability to inhibit expression of PPAR in an in vitro assay; and

selecting a candidate oxysterol compound that measurably inhibits PPAR expression.

17. The method of claim 16, wherein the screening comprises

measuring the ability of the candidate oxysterol to reduce the stimulation of PPAR expression by a PPAR agonist.

18. The method of claim 17,

wherein the PPAR agonist is contacted with the cell in the presence and in the absence of the candidate oxysterol;

and the amount of expression of PPAR is at least 3-fold less when measured in the presence of the candidate oxysterol than when measured in the absence of the candidate oxysterol.

19. The method of claim 17,

wherein the cell is a marrow stromal cell,

wherein the PPAR expression is PPAR gamma expression, and

wherein the PPAR agonist is troglitazone.