Microplate fluorescence protease assays test the inhibition of select North American snake venoms' activities with an anti-proteinase library

摘要: Snake envenomation is a relatively neglected significant world health problem, designated an orphan disease by the WHO. While often effective, antivenins are insufficient. Could another approach greatly aid inhibition of venom toxins? New fluorescent substrates for measuring protease activity in microplate assays suitable high throughput screening were tested and found reproducible with snake venom. Representative North American venoms showed strong proteinase collagenase, but weaker elastase activities. Caseinolytic inhibited nonspecific inhibitor 1,10-phenanthroline EDTA, as collagenase activity, consistent action metalloproteinases. Both general CV average 3%, Km measured above normal working conditions. Using library anti -proteinase compounds multiple revealed three agents known metalloproteinase (Actinonin, GM6001, NNGH), which incidentally supports concept that much degradative certain due to metalloproteinases activity. These results together support use assays, particularly this assay, future drug repurposing studies leading development new treatments those envenomations have major proteolytic component their pathophysiology.