Proučevanje načina vezave zaviralcev proteina toplotnega šoka Hsp90 s protivirusnim delovanjem z molekulskim sidranjem

Roštan, Klara (author), Tomašič, Tihomir (mentor)

Virusne okužbe močno ogrožajo javno zdravje celotne svetovne populacije. Protein toplotnega šoka 90 (Hsp90) ima različne vloge pri virusnih okužbah, saj je v interakciji s številnimi virusnimi proteini. Virusi za podvojevanje in širjenje potrebujejo gostiteljsko celico. Z zdravilnimi učinkovinami želimo vplivati na eno izmed faz življenjskega cikla virusa in preprečiti nadaljnje korake. Velik problem protivirusnih zdravilnih učinkovin je razvoj rezistence. V ta namen smo izvedli proučevanje zaviralcev Hsp90 kot novih spojin s protivirusnim delovanjem. Iz člankov smo povzeli ključne izsledke dosedanjih raziskav in spojine z največjim potencialom sidrali v vezavno mesto Hsp90 z uporabo treh programov. Geldanamicin in njegovi derivati so selektivni zaviralci Hsp90, ki imajo protivirusno delovanje, vendar je njihov največji problem citotoksičnost. Derivat radicikola pochonin D je 90 % zaviral razmnoževanje človeškega rinovirusa 1B v koncentraciji 2 µM. Spojina 4l je pokazala potencial za zdravljenje okužb z virusom HIV preko zaviranja Hsp90. Med 4,5,6,7-tetrahidrobenzo[1,2-d]tiazolnimi derivati je imela spojina 7 največji potencial za zdravljenje okužb z virusom hepatitisa C, saj je manj toksična kot referenčni zaviralec 17-DMAG. Med benzo[d]tiazoli je izstopala spojina 9b zaradi njene dobre vezavne afinitete na Hsp90 in delovanja proti virusu gripe brez citotoksičnosti. Gedunin se selektivno veže na N-končno domeno Hsp90 in ne povzroča toksičnosti v testiranih koncentracijah do 100 µM. S primerjavo različnih programov za sidranje smo ugotovili, da program FRED (OpenEye Scientific Software) najboljše ponovi eksperimentalno določeno vezavo zaviralcev. Spojino 17-AAG je v vezavno mesto sidral podobno kot zaviralec 17-DMAG. Iz primerjave vrednosti Kd in cenilne funkcije smo ugotovili, da ni korelacije. Rezultati sidranja so nam dali vpogled v potencialno vezavo zaviralcev, vendar bi bilo potrebno vezavno konformacijo določiti še eksperimentalno. Na ta način bi dobili primerno osnovo za nadaljnjo optimizacijo spojin za zdravljenje virusnih okužb in drugih bolezni.

• Language: Slovenian
• Keywords: Hsp90, molekulsko sidranje, protivirusno delovanje, virusne okužbe
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2022
• PID: 20.500.12556/RUL-136560
• Publication date in RUL: 11.05.2022

Secondary language

• Language: English
• Title: Binding mode studies of Hsp90 heat-shock protein inhibitors with antiviral activity by molecular docking

Abstract

Viral infections pose a serious threat to global public health. Heat shock protein 90 (Hsp90) plays many different roles in viral infections as it interacts with several viral proteins. Viruses require a host cell to replicate and spread. The goal in the treatment is to use antiviral drugs that interfere with one of the phases and prevent further steps in the life cycle of a virus. The main problem of antiviral agents is development of resistance. The aim of our study was to explore Hsp90 inhibitors as agents with antiviral activity. First, we summarized the main findings from previously published articles and then used three different programs to dock Hsp90 inhibitors with the greatest antiviral activity potential. Geldanamycin and its derivatives are selective Hsp90 inhibitors with antiviral activity, but their biggest problem is cytotoxicity. The radicicol derivative pochonin D inhibited human rhinovirus 1B replication by 90 % in a concentration of 2 µM. Compound 4l showed the potential to treat HIV infection by inhibiting Hsp90. Among the 4,5,6,7-tetrahydrobenzo[1,2-d]thiazole derivatives, compound 7 had the greatest potential for the treatment of hepatitis C virus infections, being less toxic than the reference inhibitor 17-DMAG. Among the benzo[d]thiazoles, compound 9b had the best binding affinity to Hsp90 and anti-influenza virus activity without displaying cytotoxicity. Gedunin selectively binds to the N-terminal domain of Hsp90 and does not cause toxicity at the tested concentrations up to 100 µM. Comparing different docking programs, we found that FRED (OpenEye Scientific Software) best reproduced the experimentally determined binding mode of the inhibitors. 17-AAG was docked to the binding site in a similar manner as inhibitor 17-DMAG. When comparing the Kd values and scoring functions, we found no correlation between them. The docking results gave us insight into the potential binding of the inhibitors, but it is necessary to determine the binding mode experimentally as well. This would provide us a suitable basis for further optimization of compounds for the treatment of viral infections and other diseases.

• Keywords: Hsp90, molecular docking, antiviral activity, viral infections