Methods of selecting compounds for modulation of bladder function
View Patent ↗The present invention involves an assay system and method of selecting compounds useful in the treatment of bladder instability and related bladder conditions through the activation of KCNQ potassium channels in the bladder smooth muscle.
1. A method of screening for a compound for the inhibition of urge urinary incontinence comprising:
(a) expressing a target KCNQ protein in a host cell; and
(b) detecting activation of said target KCNQ protein in the host cell; wherein the detection is performed by measuring the membrane potential of the host cell in the presence or absence of said compound and selecting the compound whose presence causes hyperpolarization in membrane potential of the host cell.
2. The method of claim 1 , wherein the host cell is an animal cell.
3. The method of claim 1 , wherein the host cell is mammalian.
4. The method of claim 1 , wherein the host cell is human.
5. The method of claim 1 , wherein the host cell is human embryonic kidney.
6. The method of claim 1 , wherein the host cell is HEK293 or a COS cell.
7. The method of claim 1 , wherein detection is performed by measuring membrane potential using an electrophysiological technique or a fluorescence technique.
8. The method of claim 1 , wherein the compound exhibits at least 2 times greater activity on a target KCNQ protein in bladder smooth muscle than on another target KCNQ protein in tissue other than bladder smooth muscle.
9. The method of claim 1 , wherein the compound exhibits at least 10 times greater activity on a target KCNQ protein in bladder smooth muscle than on another target KCNQ protein in tissue other than bladder smooth muscle.
10. The method of claim 1 , wherein the compound exhibits at least 100 times greater activity on a target KCNQ protein in bladder smooth muscle than on another target KCNQ protein in tissue other than bladder smooth muscle.
11. The method of claim 1 , wherein the compound exhibits no detectable activity on another target KCNQ protein in tissue other than bladder smooth muscle.
12. The method of claim 1 , wherein the compound exhibits at least 2 times greater activity on a target KCNQ protein in bladder smooth muscle than on a non-target KCNQ protein.
13. The method of claim 1 , wherein the compound exhibits at least 10 times greater activity on a target KCNQ protein in bladder smooth muscle than on a non-target KCNQ protein.
14. The method of claim 1 , wherein the compound exhibits at least 100 times greater activity on a target KCNQ protein in bladder smooth muscle than on a non-target KCNQ protein.
15. The method of claim 1 , wherein the compound exhibits no detectable activity on a non-target KCNQ protein.
16. The method of claim 1 , wherein the compound exhibits at least 2 times greater activity on a target KCNQ protein in bladder smooth muscle than on a protein which forms a potassium channel other than a KCNQ protein.
17. The method of claim 1 , wherein the compound exhibits at least 10 times greater activity on a target KCNQ protein in bladder smooth muscle than on a protein which forms a potassium channel other than a KCNQ protein.
18. The method of claim 1 , wherein the compound exhibits at least 100 times greater activity on a target KCNQ protein in bladder smooth muscle than on a protein which forms a potassium channel other than a KCNQ protein.
19. The method of claim 1 , wherein the compound exhibits no detectable activity on a protein which forms a potassium channel other than a KCNQ protein.
20. A method of screening for a compound for the inhibition of urge urinary incontinence comprising:
(a) expressing a target KCNQ channel in a host cell; and
(b) detecting activation of said target KCNQ channel in the host cell;
wherein the detection is performed by measuring the membrane potential of the host cell in the presence or absence of said compound and selecting the compound whose presence causes a change in membrane potential of the host cell.
21. The method of claim 20 , wherein the host cell is an animal cell.
22. The method of claim 20 , wherein the host cell is mammalian.
23. The method of claim 20 , wherein the host cell is human.
24. The method of claim 20 , wherein the host cell is human embryonic kidney.
25. The method of claim 20 , wherein the host cell is HEK293 or a COS cell.
26. The method of claim 20 , wherein detection is performed by measuring membrane potential using an electrophysiological technique or a fluorescence technique.
27. The method of claim 20 , wherein the compound exhibits at least 2 times greater activity on a target KCNQ channel in bladder smooth muscle than on another target KCNQ channel in tissue other than bladder smooth muscle.
28. The method of claim 20 , wherein the compound exhibits at least 10 times greater activity on a target KCNQ channel in bladder smooth muscle than on another target KCNQ channel in tissue other than bladder smooth muscle.
29. The method of claim 20 , wherein the compound exhibits at least 100 times greater activity on a target KCNQ channel in bladder smooth muscle than on another target KCNQ channel in tissue other than bladder smooth muscle.
30. The method of claim 20 , wherein the compound exhibits no detectable activity on a KCNQ channel in tissue other than bladder smooth muscle.
31. The method of claim 20 , wherein the compound exhibits at least 2 times greater activity on a target KCNQ channel in bladder smooth muscle than on a non-target KCNQ channel.
32. The method of claim 20 , wherein the compound exhibits at least 10 times greater activity on a target KCNQ channel in bladder smooth muscle than on a non-target KCNQ channel.
33. The method of claim 20 , wherein the compound exhibits at least 100 times greater activity on a target KCNQ channel in bladder smooth muscle than on a non-target KCNQ channel.
34. The method of claim 20 , wherein the compound exhibits no detectable activity on a non-target KCNQ channel.
35. The method of claim 20 , wherein the compound exhibits at least 2 times greater activity on a target KCNQ channel in bladder smooth muscle than on a potassium channel other than a KCNQ channel.
36. The method of claim 20 , wherein the compound exhibits at least 10 times greater activity on a target KCNQ channel in bladder smooth muscle than on a potassium channel other than a KCNQ channel.
37. The method of claim 20 , wherein the compound exhibits at least 100 times greater activity on a target KCNQ channel in bladder smooth muscle than on a potassium channel other than a KCNQ channel.
38. The method of claim 20 , wherein the compound exhibits no detectable activity on a potassium channel other than a KCNQ channel.
39. A method of screening for a compound for the inhibition of urge urinary incontinence comprising:
(a) recombinantly expressing a target KCNQ channel in a host cell;
(b) measuring membrane potential of the host cell in the presence or absence of the compound; and
(c) selecting the compound whose presence causes hyperpolarization of the host cell.
40. The method of claim 39 , wherein the host cell is an animal cell.
41. The method of claim 39 , wherein the host cell is mammalian.
42. The method of claim 39 , wherein the host cell is human.
43. The method of claim 39 , wherein the host cell is human embryonic kidney.
44. The method of claim 39 , wherein the host cell is HEK293 or a COS cell.
45. The method of claim 39 , wherein detection is performed by measuring membrane potential using an electrophysiological technique or a fluorescence technique.
46. The method of claim 39 , wherein the selected compound exhibits at least 2 times greater activity on a target KCNQ channel in bladder smooth muscle than on another target KCNQ channel in tissue other than bladder smooth muscle.
47. The method of claim 39 , wherein the selected compound exhibits at least 10 times greater activity on a target KCNQ channel in bladder smooth muscle than on another target KCNQ channel in tissue other than bladder smooth muscle.
48. The method of claim 39 , wherein the selected compound exhibits at least 100 times greater activity on a target KCNQ channel in bladder smooth muscle than on another target KCNQ channel in tissue other than bladder smooth muscle.
49. The method of claim 39 , wherein the selected compound exhibits no detectable activity on a KCNQ channel in tissue other than bladder smooth muscle.
50. The method of claim 39 , wherein the selected compound exhibits at least 2 times greater activity on a target KCNQ channel in bladder smooth muscle than on a non-target KCNQ channel.
51. The method of claim 39 , wherein the selected compound exhibits at least 10 times greater activity on a target KCNQ channel in bladder smooth muscle than on a non-target KCNQ channel.
52. The method of claim 39 , wherein the selected compound exhibits at least 100 times greater activity on a target KCNQ channel in bladder smooth muscle than on a non-target KCNQ channel.
53. The method of claim 39 , wherein the selected compound exhibits no detectable activity on a non-target KCNQ channel.
54. The method of claim 39 , wherein the selected compound exhibits at least 2 times greater activity on a target KCNQ channel in bladder smooth muscle than on a potassium channel other than a KCNQ channel.
55. The method of claim 39 , wherein the selected compound exhibits at least 10 times greater activity on a target KCNQ channel in bladder smooth muscle than on a potassium channel other than a KCNQ channel.
56. The method of claim 39 , wherein the selected compound exhibits at least 100 times greater activity on a target KCNQ channel in bladder smooth muscle than on a potassium channel other than a KCNQ channel.
57. The method of claim 39 , wherein the selected compound exhibits no detectable activity on a potassium channel other than a KCNQ channel.