Drug Repurposing of Selected FDA-approved Drugs on Inhibition of Human b-tryptase Enzymatic Activities

Ismail, Muhammad Munir (2022) Drug Repurposing of Selected FDA-approved Drugs on Inhibition of Human b-tryptase Enzymatic Activities. [Project Paper] (Submitted)

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Abstract

Dengue has been one of the consistent endemics globally, with millions of cases worldwide. Dengue hemorrhagic fever (DHF) is a more severe form of dengue infection. The main hallmark of DHF is the manifestation of severe plasma leakage, which may prove fatal if the prognosis is improperly managed. Current management of DHF includes intravenous fluid replacement therapy as well as periodic hemodynamic and vital sign reassessment during the critical phase. A recent study has reported the role of tryptase, a protease produced by mast cells, in the pathogenesis of DHF by promoting vascular permeability. Hence, a potent inhibitor against tryptase may be a potential treatment for DHF by reducing DENV-induced vascular leakage. Previously, we identified several FDA-approved drugs with strong binding affinities to human b-tryptase using a computer-assisted drug design method. The identified FDA-approved drugs are gabexate mesylate, vilanterol trifenate, argatroban, acotiamide and sunitinib. Objective: This study aims to determine the inhibitory effects of the selected FDA-approved drugs via their potency against human β-tryptase through enzymatic assay. Methodology: An optimisation assay was performed to determine the optimum concentration of β-tryptase for subsequent assays. Next, enzyme inhibition assays were performed to assess the inhibitory effects of these FDA-approved drugs against β-tryptase by co-incubating β-tryptase with different concentrations of the selected FDA-approved drugs (1, 10 and 100 µM) at 37°C. BAPNA acted as the substrate, while nafamostat mesylate was used as the positive control. Serial absorbance of the products formed was measured using a microplate reader at 405 nm for up to 72 hours at every 8 hours interval to determine the inhibitory effect of the drugs. Results: Tryptase concentration at 12.5 ng/mL was chosen as the optimum β-tryptase concentration for the subsequent enzyme inhibition assay. From the enzyme inhibition assay, gabexate mesylate was found to significantly inhibit β-tryptase activity at 10 (P = 0.0051) and 100 µM (P < 0.001). On the other hand, argatroban was found to significantly reduce β-tryptase activity only at 100 µM (P = 0.0146). Discussion: Gabexate mesylate and argatroban have been proven to significantly inhibit β-tryptase activity at different concentrations. The IC50 of both drugs should be determined in order to confirm their potency against β-tryptase. Besides, in comparison to nafamostat mesylate which is a suicide substrate that inhibits β-tryptase in an irreversible manner, both gabexate mesylate and argatroban seem to exert their inhibitory effects against β-tryptase in a reversible manner. Conclusion: Gabexate mesylate and argatroban significantly inhibit β-tryptase and thus may be potential treatments for DHF by reducing DENV-induced vascular leakage.

Item Type:	Project Paper
Faculty:	Faculty of Medicine and Health Science
Depositing User:	Ms. Nor Safa'aton Saidin
Date Deposited:	22 Aug 2023 06:34
Last Modified:	22 Aug 2023 06:34
URI:	http://psaspb.upm.edu.my/id/eprint/1351

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