High-capacity screening of arylalkylamine N-acetyltransferase inhibitors using a high-performance liquid chromatography system.
Systematic screening is a natural development of any pharmacological program. Most enzyme inhibitor screens use indirect or "aspecific" methods, such as colorimetric or fluorimetric ones. These screening methods cause quite a few false-positive and false-negative hits. In order to limit these as much as possible, we developed a methodology using a high-performance liquid chromatography (HPLC) system for the medium throughput screening of serotonin N-acetyltransferase inhibitors. The core of this screening system is (1) the dramatic shortening of the analytical time down to 100 s per run by using a high-performance analytical column (Turbo), and (2) the use of absorption as opposed to radioactivity for detection of the product of the reaction (N-acetylserotonin). This system permits the analysis of about 1,000 compounds per day to be performed with a single HPLC system. This enzymatic system was taken as an example, because the methodology can be extended to many other enzymes, particularly transferases, phosphatases, and kinases.
Ferry
G
G
Division de Pharmacologie Moléculaire et Cellulaire, Institut de Recherches Servier, Croissy-sur-Seine, France. gilles.ferry@fr.netgrs.com
Boutin
J A
JA
D001191:Arylamine N-Acetyltransferase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004795:Enzyme Stability; D006801:Humans; D020128:Inhibitory Concentration 50; D011993:Recombinant Fusion Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D015203:Reproducibility of Results; D013997:Time Factors
D004791:Enzyme Inhibitors; D011993:Recombinant Fusion Proteins; D001191:Arylamine N-Acetyltransferase