Molecules that modulate Galphaq activity and methods of treating urinary incontinence

• US 20070065883A1
• Filed: 04/14/2006
• Published: 03/22/2007
• Est. Priority Date: 01/28/2002
• Status: Abandoned Application

• Current Assignee: Bristol-Myers Squibb Company
• Original Assignee: Bristol-Myers Squibb Company
• Inventors: Fitzgerald, Kevin, Kopczynski, Jenny, Stouch, Terry, Cockett, Mark, Moore, Lisa, Doberstein, Stephen, Ramanathan, Chandra, Kindt, Rachel, Lodge, Nicholas
• Application Number:
  US11/404,725
• Publication Number:
  US 20070065883A1
• Time in Patent Office: Days
• US Class Current: 435/7.200
• CPC Class Codes: G01N 2333/726 G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH
• CPC Class Codes: G01N 2333/916 acting on ester bonds (3.1), e.g. phosphatases (3.1.3), phospholipases C or phospholipases D (3.1.4)
• CPC Class Codes: G01N 2500/00 Screening for compounds of potential therapeutic value
• CPC Class Codes: G01N 2800/341 Urinary incontinence
• CPC Class Codes: G01N 33/6893 related to diseases not provided for elsewhere

Abstract

The present invention provides methods that are useful for the treatment or prevention of smooth muscle disorders such as urinary incontinence and compounds that are useful in such methods.

Claims

1. A method for screening candidate agents that modulate interaction of RGS and Gα

q, said method comprising;

(a) contacting a screening assay system comprising a polypeptide selected from RGS and Gα

q with a candidate agent under conditions whereby, but for the presence of the agent, the system provides a reference activity; and

(b) detecting a candidate agent-biased activity of the system;

wherein a difference between the agent-biased activity and the reference activity indicates that the candidate agent modulates interaction of RGS and Gα

q.

2. The method of claim 1 wherein said assay system comprises RGS and Gα

q and said reference activity detects binding of RGS to Gα

q.

3. The method of claim 1 wherein said activity is a GAP/GTPase activity.

4. The method of claim 1 wherein said contacting in (a) comprises administering said candidate agent to cultured host cells that have been genetically engineered to express said polypeptide.

5. The method of claim 1 wherein the screening assay system is a cell-free system comprising purified polypeptide.

6. The method of claim 1 wherein the screening assay system is used in a high throughput format to screen test agents in a compound library.

7. The method of claim 1 wherein the activity is selected from carbachol-induced activity and IP3 activity.

8. The method of claim 1 wherein the reference activity comprises a measurement of muscle contraction.

9. The method of claim 8 wherein said muscle is bladder muscle strips.

10. The method of claim 1 wherein said polypeptide comprises an amino acid sequence having at least 80% sequence identity with the amino acid sequence of SEQ ID NOS:

_ (RGS &

Gα

q).

11. The method of claim 1 further comprising:

testing the candidate agent in an animal model defective for RGS or Gα

q, and detecting phenotypic changes relative to control animals.

12. A method of modulating the interaction of an RGS polypeptide and a Gα

q polypeptide comprising the step of contacting the RGS polypeptide or the Gα

q polypeptide with a compound identified by the method of claim 1.

15. The method of claim 12 wherein said assay system comprises RGS and Gα

q and reference activity detects binding of RGS to Gα

q.

16. The method of claim 12 wherein said activity is a GAP/GTPase activity.

17. The method of claim 12 wherein said contacting in (a) comprises administering said candidate agent to cultured host cells that have been genetically engineered to express said polypeptide.

18. The method of claim 12 wherein said testing in (c) is performed by administering said candidate agent to carbachol-induced bladder strips, and a reduction in muscle contraction identifies the candidate agent as capable of modulating urinary incontinence.

20. The method of claim 17 in which the agent is a chemical binding agent.

13. A method of modulating the interaction of an RGS polypeptide and a Gα

q polypeptide comprising the step of contacting the RGS polypeptide or the Gα

q polypeptide with an agent capable of modulating the interaction of the RGS polypeptide and the Gα

q.

14. A method for identifying agents that modulate urinary incontinence, said method comprising:

(a) contacting a screening assay system comprising a polypeptide selected form RGS and Gα

q with a candidate agent under conditions whereby, but for the presence of the agent, the system provides a reference activity; and

(b) detecting a candidate agent-biased activity of the system; and

(c) testing if the candidate agent that results in the biased activity detected in (b) modulates urinary incontinence.

19. A method for modulating urinary incontinence comprising contacting a vertebrate animal predetermined to have urinary incontinence with an agent that specifically binds a protein selected from the group consisting of RGS and Gα

q.