Methods of selecting compounds for modulation of bladder function

Abstract

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 selecting compounds for the treatment, prevention, or inhibition of urge urinary incontinence, bladder instability, abnormal bladder contractility, hyperactive bladder, hyperreflexive bladder, neurogenic bladder, or bladder-related sensory urgency comprising: 
 (a) expressing a target KCNQ protein in a host cell; and    (b) detecting activation of said target KCNQ protein.    
     
     
         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 the membrane potential is measured using electrophysiological techniques or fluorescence techniques.  
     
     
         8 . The method of  claim 1 , wherein compounds are selected that exhibit at least 2 times greater activity on a target KCNQ protein in bladder smooth muscle than on other target KCNQ proteins in tissue other than bladder smooth muscle.  
     
     
         9 . The method of  claim 1 , wherein compounds are selected that exhibit at least 10 times greater activity on a target KCNQ protein in bladder smooth muscle than on other target KCNQ proteins in tissue other than bladder smooth muscle.  
     
     
         10 . The method of  claim 1 , wherein compounds are selected that exhibit at least 100 times greater activity on a target KCNQ protein in bladder smooth muscle than on other target KCNQ proteins in tissue other than bladder smooth muscle.  
     
     
         11 . The method of  claim 1 , wherein compounds are selected that exhibit substantially no effect on other target KCNQ proteins in tissue other than bladder smooth muscle.  
     
     
         12 . The method of  claim 1 , wherein compounds are selected that exhibit at least 2 times greater activity on a target KCNQ protein in bladder smooth muscle than on non-target KCNQ proteins.  
     
     
         13 . The method of  claim 1 , wherein compounds are selected that exhibit at least 10 times greater activity on a target KCNQ protein in bladder smooth muscle than on non-target KCNQ proteins.  
     
     
         14 . The method of  claim 1 , wherein compounds are selected that exhibit at least 100 times greater activity on a target KCNQ protein in bladder smooth muscle than on non-target KCNQ proteins.  
     
     
         15 . The method of  claim 1 , wherein compounds are selected that exhibit substantially no effect on non-target KCNQ proteins.  
     
     
         16 . The method of  claim 1 , wherein compounds are selected that exhibit at least 2 times greater activity on a target KCNQ protein in bladder smooth muscle than on proteins which form potassium channels other than KCNQ proteins.  
     
     
         17 . The method of  claim 1 , wherein compounds are selected that exhibit at least 10 times greater activity on a target KCNQ protein in bladder smooth muscle than on proteins which form potassium channels other than KCNQ proteins.  
     
     
         18 . The method of  claim 1 , wherein compounds are selected that exhibit at least 100 times greater activity on a target KCNQ protein in bladder smooth muscle than on proteins which form potassium channels other than KCNQ proteins.  
     
     
         19 . The method of  claim 1 , wherein compounds are selected that exhibit substantially no effect on proteins which form potassium channels other than KCNQ proteins.  
     
     
         20 . A method of selecting compounds for the treatment, prevention, or inhibition of urge urinary incontinence, bladder instability, abnormal bladder contractility, hyperactive bladder, hyperreflexive bladder, neurogenic bladder, or bladder-related sensory urgency comprising: 
 (a) expressing a target KCNQ channel in a host cell; and    (b) detecting activation of said target KCNQ channel.    
     
     
         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 the membrane potential is measured using electrophysiological techniques or fluorescence techniques.  
     
     
         27 . The method of  claim 20 , wherein compounds are selected that exhibit at least 2 times greater activity on a target KCNQ protein in bladder smooth muscle than on other target KCNQ proteins in tissue other than bladder smooth muscle.  
     
     
         28 . The method of  claim 20 , wherein compounds are selected that exhibit at least 10 times greater activity on a target KCNQ protein in bladder smooth muscle than on other target KCNQ proteins in tissue other than bladder smooth muscle.  
     
     
         29 . The method of  claim 20 , wherein compounds are selected that exhibit at least 100 times greater activity on a target KCNQ protein in bladder smooth muscle than on other target KCNQ proteins in tissue other than bladder smooth muscle.  
     
     
         30 . The method of  claim 20 , wherein compounds are selected that exhibit substantially no effect on other target KCNQ proteins in tissue other than bladder smooth muscle.  
     
     
         31 . The method of  claim 20 , wherein compounds are selected that exhibit at least 2 times greater activity on a target KCNQ protein in bladder smooth muscle than on non-target KCNQ proteins.  
     
     
         32 . The method of  claim 20 , wherein compounds are selected that exhibit at least 10 times greater activity on a target KCNQ protein in bladder smooth muscle than on non-target KCNQ proteins.  
     
     
         33 . The method of  claim 20 , wherein compounds are selected that exhibit at least 100 times greater activity on a target KCNQ protein in bladder smooth muscle than on non-target KCNQ proteins.  
     
     
         34 . The method of  claim 20 , wherein compounds are selected that exhibit substantially no effect on non-target KCNQ proteins.  
     
     
         35 . The method of  claim 20 , wherein compounds are selected that exhibit at least 2 times greater activity on a target KCNQ protein in bladder smooth muscle than on proteins which form potassium channels other than KCNQ proteins.  
     
     
         36 . The method of  claim 20 , wherein compounds are selected that exhibit at least 10 times greater activity on a target KCNQ protein in bladder smooth muscle than on proteins which form potassium channels other than KCNQ proteins.  
     
     
         37 . The method of  claim 20 , wherein compounds are selected that exhibit at least 100 times greater activity on a target KCNQ protein in bladder smooth muscle than on proteins which form potassium channels other than KCNQ proteins.  
     
     
         38 . The method of  claim 20 , wherein compounds are selected that exhibit substantially no effect on proteins which form potassium channels other than KCNQ proteins.  
     
     
         39 . A method of selecting compounds for the treatment, prevention, or inhibition of urge urinary incontinence, bladder instability, abnormal bladder contractility, hyperactive bladder, hyperreflexive bladder, neurogenic bladder, or bladder-related sensory urgency comprising: 
 (a) recombinantly expressing a target KCNQ channel in a host cell;    (b) measuring the membrane potential of the host cell in the presence or absence of the a substance;    (c) selecting those compounds 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 the membrane potential is measured using electrophysiological techniques or fluorescence techniques.  
     
     
         46 . The method of  claim 39 , wherein compounds are selected that exhibit at least 2 times greater activity on a target KCNQ protein in bladder smooth muscle than on other target KCNQ proteins in tissue other than bladder smooth muscle.  
     
     
         47 . The method of  claim 39 , wherein compounds are selected that exhibit at least 10 times greater activity on a target KCNQ protein in bladder smooth muscle than on other target KCNQ proteins in tissue other than bladder smooth muscle.  
     
     
         48 . The method of  claim 39 , wherein compounds are selected that exhibit at least 100 times greater activity on a target KCNQ protein in bladder smooth muscle than on other target KCNQ proteins in tissue other than bladder smooth muscle.  
     
     
         49 . The method of  claim 39 , wherein compounds are selected that exhibit substantially no effect on other target KCNQ proteins in tissue other than bladder smooth muscle.  
     
     
         50 . The method of  claim 39 , wherein compounds are selected that exhibit at least 2 times greater activity on a target KCNQ protein in bladder smooth muscle than on non-target KCNQ proteins.  
     
     
         51 . The method of  claim 39 , wherein compounds are selected that exhibit at least 10 times greater activity on a target KCNQ protein in bladder smooth muscle than on non-target KCNQ proteins.  
     
     
         52 . The method of  claim 39 , wherein compounds are selected that exhibit at least 100 times greater activity on a target KCNQ protein in bladder smooth muscle than on non-target KCNQ proteins.  
     
     
         53 . The method of  claim 39 , wherein compounds are selected that exhibit substantially no effect on non-target KCNQ proteins.  
     
     
         54 . The method of  claim 39 , wherein compounds are selected that exhibit at least 2 times greater activity on a target KCNQ protein in bladder smooth muscle than on proteins which form potassium channels other than KCNQ proteins.  
     
     
         55 . The method of  claim 39 , wherein compounds are selected that exhibit at least 10 times greater activity on a target KCNQ protein in bladder smooth muscle than on proteins which form potassium channels other than KCNQ proteins.  
     
     
         56 . The method of  claim 39 , wherein compounds are selected that exhibit at least 100 times greater activity on a target KCNQ protein in bladder smooth muscle than on proteins which form potassium channels other than KCNQ proteins.  
     
     
         57 . The method of  claim 39 , wherein compounds are selected that exhibit substantially no effect on proteins which form potassium channels other than KCNQ proteins.  
     
     
         58 . A method of selecting a compound comprising: 
 (a) selecting compounds that do not cross the blood-brain barrier;    (b) testing those compounds for the ability to modulate a target KCNQ protein in bladder smooth muscle, and;    (c) selecting those compounds that show at least a greater ability to modulate a target KCNQ protein in bladder smooth muscle than on other target KCNQ proteins in tissue other than bladder smooth muscle.    
     
     
         59 . The method of  claim 58 , wherein the compound selected shows at least a 2 times greater ability to modulate a target KCNQ protein in bladder smooth muscle than on other target KCNQ proteins in tissue other than bladder smooth muscle.  
     
     
         60 . The method of  claim 58 , wherein the compound selected shows at least a 10 times greater ability to modulate a target KCNQ protein in bladder smooth muscle than on other target KCNQ proteins in tissue other than bladder smooth muscle.  
     
     
         61 . The method of  claim 58 , wherein the compound selected shows at least a 100 times greater ability to modulate a target KCNQ protein in bladder smooth muscle than on other target KCNQ proteins in tissue other than bladder smooth muscle.  
     
     
         62 . The method of  claim 58 , wherein the compound selected shows substantially no effect on other target KCNQ proteins in tissue other than bladder smooth muscle.  
     
     
         63 . A method of selecting a compound comprising: 
 (a) selecting compounds that do not cross the blood-brain barrier;    (b) testing those compounds for the ability to modulate a target KCNQ protein in bladder smooth muscle, and;    (c) selecting those compounds that show a greater ability to modulate a target KCNQ protein in bladder smooth muscle than on non-target KCNQ proteins.    
     
     
         64 . The method of  claim 63 , wherein the compound selected shows at least a 2 times greater ability to modulate a target KCNQ protein in bladder smooth muscle than on non-target KCNQ proteins.  
     
     
         65 . The method of  claim 63 , wherein the compound selected shows at least a 10 times greater ability to modulate a target KCNQ protein in bladder smooth muscle than on non-target KCNQ proteins.  
     
     
         66 . The method of  claim 63 , wherein the compound selected shows at least a 100 times greater ability to modulate a target KCNQ protein in bladder smooth muscle than on non-target KCNQ proteins.  
     
     
         67 . The method of  claim 63 , wherein the compound selected shows substantially no effect on non-target KCNQ proteins.  
     
     
         68 . A method of selecting a compound comprising: 
 (a) selecting compounds that do not cross the blood-brain barrier;    (b) testing those compounds for the ability to modulate a target KCNQ protein in bladder smooth muscle, and;    (c) selecting those compounds that show a greater ability to modulate a target KCNQ protein in bladder smooth muscle than proteins which form potassium channels other than KCNQ proteins.    
     
     
         69 . The method of  claim 68 , wherein the compound selected shows at least a 2 times greater ability to modulate a target KCNQ protein than proteins which form potassium channels other than KCNQ proteins.  
     
     
         70 . The method of  claim 68 , wherein the compound selected shows at least a 10 times greater ability to modulate a target KCNQ protein than proteins which form potassium channels other than KCNQ proteins.  
     
     
         71 . The method of  claim 68 , wherein the compound selected shows at least a 100 times greater ability to modulate a target KCNQ protein than proteins which form potassium channels other than KCNQ proteins.  
     
     
         72 . The method of  claim 68 , wherein the compound selected shows substantially no effect on proteins which form potassium channels other than KCNQ proteins.  
     
     
         73 . A method of selectively activating a target KCNQ protein in bladder smooth muscle of an animal for the treatment of bladder instability, comprising administering a therapeutically effective amount of a compound, which selectively activates a target KCNQ protein in bladder smooth muscle, to an animal.  
     
     
         74 . The method of  claim 73 , wherein the compound activates a target KCNQ protein in bladder smooth muscle at least 2 times greater than other target KCNQ proteins in tissue other than bladder smooth muscle.  
     
     
         75 . The method of  claim 73 , wherein the compound activates a target KCNQ protein in bladder smooth muscle at least 10 times greater than other target KCNQ proteins in tissue other than bladder smooth muscle.  
     
     
         76 . The method of  claim 73 , wherein the compound activates a target KCNQ protein in bladder smooth muscle at least 100 times greater than other target KCNQ proteins in tissue other than bladder smooth muscle.  
     
     
         77 . The method of  claim 73 , wherein the compound activates a target KCNQ protein in bladder smooth muscle and shows substantially no effect on other target KCNQ proteins in tissue other than bladder smooth muscle.  
     
     
         78 . The method of  claim 73 , wherein the compound activates a target KCNQ protein in bladder smooth muscle at least 2 times greater than on non-target KCNQ proteins.  
     
     
         79 . The method of  claim 73 , wherein the compound activates a target KCNQ protein in bladder smooth muscle at least 10 times greater than on non-target KCNQ proteins.  
     
     
         80 . The method of  claim 73 , wherein the compound activates a target KCNQ protein in bladder smooth muscle at least 100 times greater than on non-target KCNQ proteins.  
     
     
         81 . The method of  claim 73 , wherein the compound activates a target KCNQ protein in bladder smooth muscle and shows substantially no effect on non-target KCNQ proteins.  
     
     
         82 . The method of  claim 73 , wherein the compound activates a target KCNQ protein in bladder smooth muscle at least 2 times greater than proteins which form potassium channels other than KCNQ proteins.  
     
     
         83 . The method of  claim 73 , wherein the compound activates a target KCNQ protein in bladder smooth muscle at least 10 times greater than proteins which form potassium channels other than KCNQ proteins.  
     
     
         84 . The method of  claim 73 , wherein the compound activates a target KCNQ protein in bladder smooth muscle at least 100 times greater than proteins which form potassium channels other than KCNQ proteins.  
     
     
         85 . The method of  claim 73 , wherein the compound activates a target KCNQ protein in bladder smooth muscle and shows substantially no effect on proteins which form potassium channels other than KCNQ proteins.