Trimethyl Lock A Stable Chromogenic Substrate for Esterases 英文参考文献.docVIP

Molecules 2008, 13, 204–211 molecules ISSN 1420-3049 ? 2008 by MDPI /molecules Full Paper Trimethyl Lock: A Stable Chromogenic Substrate for Esterases Michael N. Levine 1, Luke D. Lavis 2 and Ronald T. Raines 1,2,* 1 Department of Biochemistry, University of Wisconsin–Madison, 433 Babcock Drive, Madison, WI 53706-1544, USA 2 Department of Chemistry, University of Wisconsin–Madison, 1101 University Avenue, Madison, WI 53706-1322, USA * Author to whom correspondence should be addressed; E-mail: rtraines@ Received: 19 January 2008 / Accepted: 30 January 2008 / Published: 31 January 2008 Abstract: p-Nitrophenyl acetate is the most commonly used substrate for detecting the catalytic activity of esterases, including those that activate prodrugs in human cells. This substrate is unstable in aqueous solution, limiting its utility. Here, a stable chromogenic substrate for esterases is produced by the structural isolation of an acetyl ester and p-nitroaniline group using a trimethyl lock moiety. Upon ester hydrolysis, unfavorable steric interactions between the three methyl groups of this o-hydroxycinnamic acid derivative encourage rapid lactonization to form a hydrocoumarin and release p-nitroaniline. This “prochromophore” could find use in a variety of assays. Keywords: enzyme catalysis, chromogenic substrate, p-nitrophenyl acetate, trimethyl lock Introduction Prodrug strategies have been employed to improve the properties of potential small-molecule chemotherapeutic agents, including their solubility, stability, organ selectivity, duration of action, and bioavailability [1, 2]. Recently, our research group reported on the use of the “trimethyl lock” prodrug strategy as the basis for a new class of latent fluorophores [3–5]. The trimethyl lock is an o-hydroxycinnamic acid derivative in which severe steric repulsion between three methyl groups leads to r