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 inhibitors, from a library of typical enzyme inhibitors. By screening 10,000 culture broths of microorganism (actinomycetes, fungi, and bacteria) an actinomycete strain, RK99-A234, was found to have haparanase inhibitory activity. RK-682 was identified in the fermentation broths as a heparanase inhibitors, IC50 = 17 μM.
ASJC Scopus subject areas
- Drug Discovery