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A chemical library was screened for inhibition of lethal factor (LF), one of anthrax toxin, by in vitro and in vivo analysis. Lethal factor has proteolytic activity to cleave MEK1 protein. This proteolytic activity was used for in vitro screening of chemicals by using GST-MEK1 as a substrate of lethal factor. The optimum metal ion concentrations for high proteolytic activity of LF were 50 M Caand 5 mM Mg. 2,6-Dimercapto-3,5-diphenyl-thiopyran4-one (FBS00831) was a strong inhibitor of LF and showed IC50 = 100 M by in vitro analysis. RAW 264.7 cell line and DMSO concentrations were established to 12.5% and 4% respectively. After treatment with protective antigen, lethal factor was endocytosed into RAW 264.7 cells, which were treated by FBS00831 for in vivo assay. 1.2 mM FBS00831 recovered 67% cell viability as compare to 80% recovery by GM6001 (known metalloprotease inhibitor) at 0.4 mM concentration.

In Vitro Screening and In Vivo Analysis of Anthrax Lethal Factor Inhibitor