Use of azetidinone compounds
AssigneeSchering Corporation
InventorsScott W. Altmann, Duane A. Burnett, Harry R. Davis JR., Michael P. Graziano, Maureen Laverty, Xiaorui Yao
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Abstract
The use of azetidinone compounds that are inhibitors of cholesterol absorption as tools for discovering and characterizing proteins involved in trafficking or absorption of cholesterol and/or cholesteryl esters in biological systems is presented. These compounds can serve as tools for competitive binding assays to discover and characterize other chemical agents useful as cholesterol absorption inhibitors. New compounds of the present invention are also highly efficacious inhibitors of cholesterol absorption.
Claims28
A method for identifying a protein involved in cholesterol absorption in a biological system comprising the steps of:
- (a) providing a cDNA expression library capable of expressing a protein involved in cholesterol absorption in a biological system, said cDNA expression library comprising a plurality of cells capable of expressing different cDNAs;
- (b) screening said expression library by incubating cells from said library with a fluorescent cholesterol absorption inhibitor;
- (c) after step (b), identifying the cell or cells in said library that display the greatest amount of fluorescence;
- and
- (d) identifying the protein associated with the cDNA expressed by said cell or cells displaying the greatest amount of fluorescence.
A method for assaying inhibitory agents for activity against cholesterol absorption comprising the steps of:
- providing a cell capable of binding a fluorescent cholesterol absorption inhibitor;
- contacting said cell with a candidate inhibitory agent in the presence of said fluorescent cholesterol absorption inhibitor;
- and
- measuring the inhibition of the fluorescence of said cell.
A method for identifying inhibitory agents which inhibit the absorption of cholesterol into a cell membrane, said method comprising the steps of:
- (a) combining a fluorescent cholesterol absorption inhibitor, said cell membrane and a candidate inhibitory agent, under conditions wherein, but for the presence of said inhibitory agent, said fluorescent cholesterol absorption inhibitor is bound to the membrane;
- and
- (b) detecting the relative presence or absence of fluorescent cholesterol absorption inhibitor absorption bound to the membrane,
- wherein a relative absence of fluorescent cholesterol absorption inhibitor absorption indicates that said candidate inhibitory agent is an inhibitory agent which inhibits cholesterol absorption into the membrane.
A method for identifying inhibitory agents which inhibit the absorption of cholesterol, said method comprising the steps of:
- (a) combining a labeled cholesterol absorption inhibitor, a cell expressing the scavenger receptor type B, class I (SR-BI) and a candidate inhibitory agent, under conditions wherein, but for the presence of said inhibitory agent, said labeled cholesterol absorption inhibitor binds to SR-BI;
- and
- (b) detecting the relative presence or absence of labeled cholesterol absorption inhibitor absorption bound to SR-BI,
- wherein a relative absence of labeled cholesterol absorption inhibitor absorption indicates that said candidate inhibitory agent is an inhibitory agent which inhibits SR-BI-mediated cellular cholesterol absorption.
A fluorescent cholesterol absorption inhibitor of formula I or II:
- 20
- wherein R comprises a fluorescent moiety.
Dependencies are inferred from the claim text.