High-capacity screening of arylalkylamine N-acetyltransferase inhibitors using a high-performance liquid chromatography system

Systematic screening is a natural development of any pharmacological progra m. 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 muc h as possible, we developed a methodology using a high-performance liquid c hromatography (HPLC) system for the medium throughput screening of serotoni n 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 usi ng a high-performance analytical column (Turbo), and (2) the use of absorpt ion as opposed to radioactivity for detection of the product of the reactio n (N-acetylserotonin). This system permits the analysis of about 1,000 comp ounds per day to be performed with a single HPLC system. This enzymatic sys tem was taken as an example, because the methodology can be extended to man y other enzymes, particularly transferases, phosphatases, and kinases.