Rapid screening of enzymes for the enzymatic hydrolysis of chiral esters in drug discovery.
Thirty-five enzymes were rapidly screened for their ability to selectively hydrolyze chiral esters to their corresponding carboxylic acids for the efficient generation of chiral intermediates in drug discovery. Optimization of the enzymatic reactions at various incubation times was performed using a robotic liquid handler. Enantiomeric pairs of chiral esters and carboxylic acids were then analyzed simultaneously by chiral GC/MS in a single analysis. This analytical approach is particularly useful for compounds that do not possess a conjugated chromophore or are volatile and difficult to analyze by chiral HPLC/UV or HPLC/MS. The resulting data was used to determine enantiomeric excesses and percent conversions to the desired enantiomer of the carboxylic acid for the selection of efficient enzymes for bioconversions in drug discovery in a pharmaceutical company.
Petucci
Chris
C
Discovery Analytical Chemistry, Chemical Technologies, Chemical and Screening Sciences, Wyeth Research, Collegeville, PA 19426, USA. petuccc@wyeth.com
Di
Li
L
McConnell
Oliver
O
D000818:Animals; D002626:Chemistry, Pharmaceutical / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D015195:Drug Design; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D006868:Hydrolysis; D007700:Kinetics; D013058:Mass Spectrometry / Q000379:methods; D008956:Models, Chemical; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D013237:Stereoisomerism; D013552:Swine; D013678:Technology, Pharmaceutical / Q000379:methods; D013997:Time Factors