High-throughput screening for human collagenase 1 inhibitors.
OBJECTIVE
To establish a high-throughput method for inhibitor screening using a recombinant collagenase catalytic domain.


METHODS
Human collagenase 1 catalytic domain protein was expressed in E coli and used for screening a set of 2720 compounds in a high-throughput fashion.


RESULTS
The screening was accomplished within 2 h and 10 min with consumption of each compound at 4 micrograms. Sixty-six compounds were identified with > 60% inhibitory activity at 20 mg/L, among which 44 compounds were confirmed by subsequent testing at multiple concentrations. The most potent compound showed an IC50 at 4.3 mumol/L, and there were total 15 compounds with IC50 less than 20 mumol/L.


CONCLUSIONS
The high-throughput method using the recombinant collagenase is fast, effective and practical in identifying inhibitors.
Ye
Q Z
QZ
National Centers for Drug Screening, Shanghai Institute of Materia Medica, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
Nan
F J
FJ
Hu
L Y
LY
Qian
Z
Z
Qian
J
J
D017364:Collagenases / Q000096:biosynthesis; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004926:Escherichia coli / Q000378:metabolism; D020128:Inhibitory Concentration 50; D061965:Matrix Metalloproteinase Inhibitors; D011994:Recombinant Proteins / Q000096:biosynthesis
D061965:Matrix Metalloproteinase Inhibitors; D011994:Recombinant Proteins; D017364:Collagenases; C417249:collagenase 1