- JAPAN ANTIBIOTICS RESEARCH ASSOCIATION
- The Journal of Antibiotics (ISSN:00218820)
- vol.57, no.2, pp.136-142, 2004-02-25 (Released:2009-01-27)
In this paper we describe the establishment of an efficient visual method for screening heparanase inhibitors, and we present the results of screening 10, 000 microbial culture broths. Heparanase-overexpressing stable clones of the human hepatocellular carcinoma HepG2 cells were established and used as an enzyme source. Digestion of heparan sulfate (HS) was detected using novel HS-containing tablets or SDS-polyacrylamide gel electrophoresis. This method was able to find suramin, a known heparanase inhibitor, from a library of typical enzyme inhibitors. By screening 10, 000 culture broths of microorganisms (actinomycetes, fungi, and bacteria) an actinomycete strain, RK99-A234, was found to have heparanase inhibitory activity. RK-682 was identified in the fermentation broth as a heparanase inhibitor, IC50=17μM.