Method of identifying a PPARgamma-agonist compound having a decreased likelihood of inducing dose-dependent peripheral edema

• US 7,482,124 B2
• Filed: 07/07/2006
• Issued: 01/27/2009
• Est. Priority Date: 07/08/2005
• Status: Active Grant

• Current Assignee: Bristol-Myers Squibb Company
• Original Assignee: Bristol-Myers Squibb Company
• Inventors: Geese, William J., Delmonte, Terrye Aigeldinger, Ranade, Koustubh
• Primary Examiner(s): Landsman; Robert
• Application Number:
  US11/483,290
• Publication Number:
  US 20070009945A1
• Time in Patent Office: 935 Days
• Field of Search: None
• US Class Current: 435/6.14
• CPC Class Codes: C07K 14/57536 Endothelin, vasoactive intestinal contractor [VIC]
• CPC Class Codes: C07K 14/70571 for neuromediators, e.g. serotonin receptor, dopamine receptor
• CPC Class Codes: C12N 9/6486 Renin (3.4.23.15)
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/106 Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
• CPC Class Codes: C12Q 2600/136 Screening for pharmacological compounds
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: C12Q 2600/178 miRNA, siRNA or ncRNA
• CPC Class Codes: C12Y 304/23015 Renin (3.4.23.15)
• CPC Class Codes: G01N 2333/70567 Nuclear receptors, e.g. retinoic acid receptor [RAR], RXR, nuclear orphan receptors
• CPC Class Codes: G01N 2500/10 involving cells
• CPC Class Codes: G01N 33/5023 on expression patterns

Abstract

The invention provides novel polynucleotides and polypeptides associated with the incidence of PPAR-agonist induced edema. The invention also provides polynucleotide fragments corresponding to the genomic and/or coding regions of these polynucleotides which comprise at least one polymorphic locus per fragment. Allele-specific primers and probes which hybridize to these regions, and/or which comprise at least one polymorphic locus are also provided. The polynucleotides, primers, and probes of the present invention are useful in phenotype correlations, medicine, and genetic analysis. Also provided are vectors, host cells, antibodies, and recombinant and synthetic methods for producing said polynucleotides and/or polypeptides. The invention further relates to diagnostic and therapeutic methods for applying these novel polynucleotides and polypeptides to the diagnosis, treatment, and/or prevention of various diseases and/or disorders, particularly PPAR-agonist induced edema or related indications. The invention further relates to screening methods for identifying agonists of PPAR proteins with decreased risk of inducing peripheral edema in patients.

Claims

1. A method of identifying a PPARgamma-agonist compound having a decreased likelihood of inducing dose-dependent peripheral edema in a patient comprising the step of:

(a) incubating mammalian cells that endogenously express renin with a test compound; and

(b) measuring the level of induced expression of renin mRNA in response to said test compound;

wherein a decreased level of induced renin mRNA expression by said test compound relative to a reference compound is indicative of a decreased risk of said test compound inducing dose-dependent peripheral edema in a patient.

2. The method according to claim 1, wherein said cells are human CALU-6 cells.

3. The method according to claim 1, further comprising the step of measuring the level of induced expression of endothelin-1 mRNA in response to said test compound wherein said cells endogenously express endothelin-1;

and

wherein an increased level of induced endothelin-1 expression by the test compound relative to a reference compound is indicative of a decreased risk of said compound inducing dose-dependent peripheral edema in a patient.

4. The method according to claim 3, wherein said cells are human CALU-6 cells.

5. The method according to claim 1, wherein the measurement of human renin mRNA expression is performed using the polymerase chain reaction.

6. The method according to claim 1, wherein the measurement of human renin mRNA expression is performed using the quantitative polymerase chain reaction.

7. The method according to claim 1, wherein the measurement of human renin mRNA expression is performed using the reverse-transcribed polymerase chain reaction.

8. The method according to claim 1, wherein the measurement of human renin mRNA expression is performed using microarrays.

9. The method according to claim 1, wherein the measurement of human renin mRNA expression is performed using mass spectrometry.

10. A method according to claim 1, wherein said renin mRNA encodes the renin polypeptide with one amino acid substitution.

11. A method according to claim 3, wherein said endothelin-1 mRNA encodes the endothelin-1 polypeptide with one amino acid substitution.

12. A method of identifying whether a PPARgamma-agonist compound has an increased likelihood of inducing dose-dependent peripheral edema in a patient comprising the step of:

(a) incubating mammalian cells that endogenously express renin with a test compound; and

(b) measuring the level of induced expression of renin mRNA in response to said test compound;

wherein an increased level of induced renin mRNA expression by said test compound relative to a reference compound is indicative of an increased risk of said test compound inducing dose-dependent peripheral edema in a patient, whereas a decreased level of induced renin mRNA expression by said test compound relative to a reference compound is indicative of a decreased risk of said test compound inducing dose-dependent peripheral edema in a patient.

13. The method according to claim 12, wherein said cells are human CALU-6 cells.

14. The method according to claim 12, further comprising the step of measuring the level of induced expression of endothelin-1 mRNA in response to said test compound, wherein said cells endogenously express endothelin-1;

wherein a decreased level of induced endothelin-1 expression by the test compound relative to a reference compound is indicative of an increased risk of said compound inducing dose-dependent peripheral edema in a patient, whereas an increased level of induced endothelin-1 expression by the test compound relative to a reference compound is indicative of a decreased risk of said compound inducing dose-dependent peripheral edema in a patient.

15. The method according to claim 14, wherein said cells are human CALU-6 cells.

16. A kit for identifying a PPAR gamma-agonist compound having a decreased likelihood of inducing dose-dependent peripheral edema in a patient comprising:

(a) a test compound;

(b) a means for incubating mammalian cells that endogenously express renin with said test compound;

(c) a means for measuring the level of induced expression of renin mRNA in response to said test compound;

wherein a decreased level of induced renin mRNA expression by said test compound relative to a reference compound is indicative of a decreased risk of said test compound inducing dose-dependent peripheral edema in a patient.

17. The kit according to claim 16, further comprising a means for measuring the level of induced expression of endothelin-1 mRNA in response to said test compound, wherein said cells endogenously express endothelin-1;

and

wherein an increased level of induced endothelin-1 expression by the test compound relative to a reference compound is indicative of a decreased risk of said compound inducing dose-dependent peripheral edema in a patient.