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Science 5 March 1993:
Vol. 259. no. 5100, pp. 1445 - 1450
DOI: 10.1126/science.8451640
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Articles

Science, Vol 259, Issue 5100, 1445-1450
Copyright © 1993 by American Association for the Advancement of Science

articles

Structure-based discovery of inhibitors of thymidylate synthase

BK Shoichet, RM Stroud, DV Santi, ID Kuntz, and KM Perry

Department of Pharmaceutical Chemistry, University of California, San Francisco 94143.

A molecular docking computer program (DOCK) was used to screen the Fine Chemical Directory, a database of commercially available compounds, for molecules that are complementary to thymidylate synthase (TS), a chemotherapeutic target. Besides retrieving the substrate and several known inhibitors, DOCK proposed putative inhibitors previously unknown to bind to the enzyme. Three of these compounds inhibited Lactobacillus casei TS at submillimolar concentrations. One of these inhibitors, sulisobenzone, crystallized with TS in two configurations that differed from the DOCK-favored geometry: a counterion was bound in the substrate site, which resulted in a 6 to 9 angstrom displacement of the inhibitor. The structure of the complexes suggested another binding region in the active site that could be exploited. This region was probed with molecules sterically similar to sulisobenzone, which led to the identification of a family of phenolphthalein analogs that inhibit TS in the 1 to 30 micromolar range. These inhibitors do not resemble the substrates of the enzyme. A crystal structure of phenolphthalein with TS shows that it binds in the target site in a configuration that resembles the one suggested by DOCK.


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