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Katherine Hu, 1, 2 Zhaowen Zhu, 1, 2 Meleah M Mathahs, 1, 2 Huy Tran, 1, 2 Jerry Bommer, 3, 4 Charles A Testa, 5 Warren N Schmidt 1, 2 1Department of Internal Medicine and Research Service, Veterans Affairs Medical Center, Iowa City, IA 52246, USA; 2Department of Internal Medicine of the Roy G. And Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA; 3Frontier Scientific, Logan, UT 84321, USA; 4Echelon Biosciences Inc, Salt Lake City, UT 84108, USA; 5Curza, Provo, UT and Cambridge, MA, USACorrespondence: Warren N SchmidtDepartment of Internal Medicine, University of Iowa Hospitals and Clinics, 200 Hawkins Drive, 4553 JCP, Iowa City, IA 52242, USATel +1 319 353-7048Fax +1 319 356-7918Email warren-schmidt@uiowa.eduBackground: Antiviral actions of tetrapyrroles have been described in a number of systems. Our goal was to evaluate antagonism of the HCV NS3-4A protease by a variety of common porphyrins and characterize structure–activity relationships that may be useful for future drug design of HCV and related Flaviviruses.Methods: Using fluorometric assays, common metalloprotoporphyrins (MPP) all inhibited NS3-4A protease with IC 50 values in low micromolar ranges [CoPP (1.4 μM) < ZnPP = MnPP = SnPP < CuPP < FePP (6.5 μM) = protoporphyrin].Results: Lineweaver–Burk plots confirmed that MPP: NS3 inhibition was basically competitive. All tested MPPs inhibited HCV genotype 1A, 1B, 2A and 3A recombinant proteases with the same fidelity suggesting wide antagonistic capabilities. However, when the MPPs were tested in cellular incubations with HCV replicons only Zn, Fe and free-base protoporphyrin showed comparable EC 50 and IC 50 values suggesting that there may be critical differences in MPP uptake and intracellular availability. Meso, deutero, and isohematoporphyrin derivatives, with or without metal substitution, all showed less anti-protease and antiviral activities as compared to protoporphyrins, suggesting that the planar, vinyl side chains are important for protease active site binding. MPPs were also active against three common protease mutants (T54A, A156T, and V36M) with equivalent or better IC 50 values as compared to wild type enzyme.Conclusion: These findings document the versatility of MPPs as antiviral agents with an expanded sensitivity for HCV genotypes and resistance to some common viral mutations. The results also suggest that further study of MPP structure and function will be useful for the development of new antiviral agents.Keywords: porphyrins, antiviral agents, hepatitis C virus, hepatitis C genotype |
