High-throughput screening for human collagenase 1 inhibitors

AIM: To establish a high-throughput method for inhibitor screening using a recombinant collagenase catalytic domain. METHODS: Human collagenase 1 cata lytic domain protein was expressed in E coli and used for screening a set o f 2720 compounds in a high-throughput fashion. RESULTS: The screening was a ccomplished within 2 h and 10 min with consumption of each compound at 4 mu g. Sixty-six compounds were identified with > 60 % inhibitory activity at 2 0 mg/L, among which 44 compounds were confirmed by subsequent testing at mu ltiple concentrations. The most potent compound showed an IC50 at 4.3 mu mo l/L, and there were total 15 compounds with IC50 less than 20 mu mol/L. CON CLUSION: The high-throughput method using the recombinant collagenase is fa st, effective and practical in identifying inhibitors.