Methods for the screening of non-recombinant cell lines capable of expressing a single PDE4 isoenzyme and for the screening of PDE4 inhibitors
AssigneeWarner-Lambert Company
InventorCorinne Szilagyi
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Abstract
The invention concerns a method for the screening of a non-recombinant cell line capable, under appropriate conditions, of exhibiting an upregulated expression of a target protein, preferably an isoenzyme of PDE4, more preferably PDE4A. The invention also concerns methods for the screening of a candidate molecule that modulates the expression or the activity of human phosphodiesterase 4A. The candidate molecules selected by the screening methods of the invention are potentially useful as therapeutic molecules for diseases caused or regulated by cyclic nucleotide-modulated transduction mechanisms, such as airway disorders like asthma.
Claims7
A method for determining the selectivity of a candidate molecule for a human phosphodiesterase 4A (PDE 4A), said method comprising:
- a) incubating a non-recombinant cell line culture having a significantly upregulated expression of a PDE 4A and a substantially downregulated expression of other PDE4 isoenzymes by treating with a concentration of a phorbol ester sufficient to significantly increase PDE 4A production in said cell line.
- b) adding said candidate compound to said cell line culture;
- and
- c) quantifying the variation of cyclic-3′,5′-adenosine monophosphate (cAMP) contents in said culture.
Dependencies are inferred from the claim text.