PHARMACEUTICAL COMPOSITIONS FOR SUPPRESSING B-AMYLOID PRODUCTION
To provide pharmaceutical compositions for suppressing β-amyloid production on the basis of the cyclin-dependent kinase inhibitory activity. When a substance having a cyclin-dependent kinase inhibitory activity was allowed to contact with nerve cells, the β-amyloid production was suppressed.
1 - 10 . (canceled)
11 . A method for identifying a substance having β-amyloid production suppressing action, comprising:
(a) contacting a substrate with cyclin-dependent kinase (Cdk5) and Cdk5 inhibitor in the presence of ATP, and
(b) terminating the enzyme reaction and measuring the amount of phosphorylation of the substrate.
12 . A method for identifying a substance having β-amyloid production suppressing action, comprising:
(a) contacting a substrate with cyclin-dependent kinase (Cdk5) and Cdk5 inhibitor in the presence of ATP,
(b) terminating the enzyme reaction and measuring the amount of phosphorylation of the substrate,
(c) contacting a cell with the Cdk5 inhibitor, and
(d) determining whether said Cdk5 inhibitor inhibits APP phosphorylation in the cell thereby identifying a compound having β-amyloid production suppressing action.
13 . The method of claim 11 or 12 , wherein said substrate is APP, histone H1 or their biotinylated forms.
14 . A method of claim 12 , wherein said ATP is isotope-labeled ATP.
15 . The method of claim 12 , wherein said inhibition of APP phosphorylation is determined by western blot using an antibody specific for phosphorylated APP.
16 . The method of claim 12 , wherein said cell does not proliferate.
17 . The method claim 16 , wherein said cell is a nerve cell.
18 . A method for identifying a substance having β-amyloid production suppressing action, comprising:
(a) contacting a substrate with cyclin-dependent kinase (Cdk5) and a test compound in the presence of ATP, and
(b) terminating the enzyme reaction and measuring the amount of phosphorylation of the substrate.
19 . A method for identifying a substance having β-amyloid production suppressing action, comprising:
(a) contacting a substrate with cyclin-dependent kinase (Cdk5) and a test compound in the presence of ATP,
(b) terminating the enzyme reaction and measuring the amount of phosphorylation of the substrate,
(c) contacting a cell with the test compound, and
(d) determining whether said test compound inhibits APP phosphorylation in the cell thereby identifying a compound having β-amyloid production suppressing action.
20 . The method of claim 18 or 19 , wherein said substrate is APP, histone H1 or their biotinylated form.
21 . The method of claim 19 , wherein said ATP is isotope-labeled ATP.
22 . The method of claim 19 , wherein said inhibition of APP phosphorylation is determined by western blot using an antibody specific for phosphorylated APP.
23 . The method of claim 19 , wherein said cell does not proliferate.
24 . The method of claim 23 , wherein said cell is a nerve cell.