A Recombinant Human Stromelysin Catalytic Domain Identifying Tryptophan Derivatives as Human Stromelysin Inhibitors

Abstract

The human stromelysin catalytic domain (SCD) has been expressed in Escherichia coli and purified to homogeneity (Ye et al. Biochemistry 1992, 31, 11231). We have used this recombinant SCD for inhibitor screening and identified tryptophan derivatives as competitive inhibitors of SCD. Both Cbz-L-Trp-OH (1, IC50 2.5 μM, Ki 2.1 μM) and Boc-L-Trp-OH (3, IC50 10 μM, Ki 8 μM) showed good inhibitory activity. Modification at the indole nitrogen with formyl or mesitylene-2-sulfonyl group (16, IC50 34 μM. Ki 28 μM; 17, IC50 63 μM, Ki 52 μM) showed reduced activity. The amide Cbz-L-Trp-NH2 (13) was not active, but esters Cbz-L-Trp-OSu (14, IC50 13 μM, Ki 11 μM) and Boc-L-Trp-OSu (15, IC50 102 μM, Ki 84 μM) showed activity. Aromatic amino acid derivatives Cbz-L-Tyr-OH (18, IC50 24 μM, Ki 20 μM) and Cbz-L-Phe-OH (26, IC50 40 μM, Ki 33 μM) were also active, but other amino acid derivatives had no activity. Although Cbz-d-Trp-OH (2, IC50 86 μM, Ki 71 μM) was active, the L-configuration is consistently preferred for inhibitory activity. Some of the SCD inhibitors were tested on full-length human stromelysin purified from cultured human cells, and they showed the same potency rank order. These results demonstrate the usefulness of recombinant DNA technology in generating the authentic human protein with improved properties for drug discovery. © 1994, American Chemical Society. All rights reserved.