Establishment of A Drug-Screening System for the Omsk Hemorrhagic Fever Virus NS2B-NS3 Protease and Inhibitor Selection
Omsk hemorrhagic fever virus(OHFV)is a tick-borne flavivirus found primarily in Siberian(Russia).It infects humans through tick bites or contact with infectious materials.Symptoms of infection include headache,cough,fever,and bleeding.Specific drugs targeting this virus are not available.The translated genome of OHFV forms polyproteins,among which the NS3 protease cleaves polyproteins to fulfill their respective roles in the viral replication cycle.The hydrophilic region of NS2B acts as a cofactor to maintain the hydrolytic activity of NS3 protease,having an indispensable role in the viral lifecycle.We aimed to develop small-molecule inhibitors of Omsk hemorrhagic fever by targeting the NS2B-NS3 protease.The NS2B-NS3 of OHFV protease was expressed using the Escherichia coli expression system to obtain highly pure and homogeneous protein.Simultaneously,a drug screening system was established to conduct inhibitor screening.The final drug-screening system consisted of Tris(20 mmol/L)and 20％glycerol at pH 8.5,with a final protein concentration of 2 μmol/L and a substrate concentration of 100 μmol/L.Utilizing this system for inhibitor screening,the compound zinc pyrithione,was identified with percent inhibition up to 99％against NS2B-NS3 protease of OHFV.The half-maximal inhibitory concentration of this inhibitor was determined,and its inhibitory type was identified as reversible.Zinc pyrithione shows promise to be a lead compound against Omsk hemorrhagic fever virus,providing a research basis for drug development targeting this virus.
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