Pharmacologic Method Of Lowering Cholesterol Production

Abstract

The present invention provides a screening method for identifying test agents that modulate cell membrane cholesterol activity. The modulating activity of the test agents may be measured using lytic compounds, which cause cell lysis or increases in cell permeability in response to cell membrane cholesterol levels. The invention further provides therapeutic agents that are identified using the screening method. The therapeutic agents either increase or decrease the cell membrane cholesterol activity, cholesterol concentration and/or both in vivo and/or in vitro.

1 . A method of screening compounds for cholesterol modulating activity, comprising: 
 (a) contacting one or more test agents with one or more cells; and    (b) determining whether the one or more test agents has an effect on cholesterol activity, cholesterol concentration or both in a membrane of the one or more cells.    
     
     
         2 . The method of  claim 1 , wherein (b) comprises contacting the one or more cells with a lytic compound, wherein the lytic compound causes perforation or lyses of the membrane of the one or more cells when the cholesterol activity, cholesterol concentration or both of the membrane of the one or more cells reaches a level at or above a threshold cholesterol level.  
     
     
         3 . The method of  claim 1 , further comprising performing (a) and (b) one or more times with different test agents.  
     
     
         4 . The method of  claim 3 , wherein the different test agents are screened simultaneously.  
     
     
         5 . The method of  claim 1 , further comprising: 
 (c) increasing or decreasing the cholesterol content of the membrane by contacting the one or more cells with a cholesterol modulating compound.    
     
     
         6 . The method of  claim 5  wherein (c) is performed prior to (a), simultaneous with (a) or subsequent to (a).  
     
     
         7 . The method of  claim 5 , wherein the cholesterol modulating compound comprises a cyclodextrin or cyclodextrin derivative.  
     
     
         8 . The method of  claim 2 , wherein the lytic compound comprises a polyene antibiotic.  
     
     
         9 . The method of  claim 2 , wherein the lytic compound comprises a lysophosphatide or cholesterol oxidase.  
     
     
         10 . The method of  claim 2 , wherein the lytic compound comprises a bacterial toxin.  
     
     
         11 . The method of  claim 1 , wherein the one or more cells comprise one or more eukaryotic cells.  
     
     
         12 . The method of  claim 11 , wherein the one or more eukaryotic cells comprise one or more mammalian cells.  
     
     
         13 . The method of  claim 11 , wherein the one or more cells comprise one or more red blood cells.  
     
     
         14 . The method of  claim 11 , wherein the one or more cells comprise one or more fibroblasts.  
     
     
         15 . The method of  claim 11 , wherein the one or more cells comprise one or more human cells.  
     
     
         16 . The method of  claim 1 , wherein the one or more cells have vigorous cholesterol homeostasis.  
     
     
         17 . The method of  claim 1 , wherein (b) comprises measuring the effect, if any, the test agent has on the cholesterol activity, cholesterol concentration or both in the membrane of the one or more cells.  
     
     
         18 . The method of  claim 1 , wherein (b) comprises measuring the permeability of the membrane of the one or more cells or the turbidity of the one or more cells.  
     
     
         19 . The method of  claim 1 , wherein the one or more cells are in vitro.  
     
     
         20 . The method of  claim 1 , wherein the one or more cells are in vivo.  
     
     
         21 . The method of  claim 1 , wherein (b) comprises measuring the cholesterol activity, cholesterol concentration or both in a plasma membrane of the one or more cells.  
     
     
         22 - 25 . (canceled)  
     
     
         26 . A method of modulating a cholesterol level of a cell comprising contacting one or more cells with an effective amount of octanol, ceramide, diglyceride, lysophosphosphatidyl choline, or combinations thereof, thereby increasing or decreasing the cholesterol level of the one or more cells.  
     
     
         27 . The method of  claim 26  wherein the one or more cells are in vivo.  
     
     
         28 . (canceled)