Methodology for screening of HIV inhibitors applicable for in silico designed molecules and empirical testing of traditional medicinal materials

0506507 - ÚOCHB 2020 RIV NL eng C - Konferenční příspěvek (zahraniční konf.)
Rumlová, Michaela - Spiwok, V. - Ruml, T. - Hadravová, Romana
Methodology for screening of HIV inhibitors applicable for in silico designed molecules and empirical testing of traditional medicinal materials.
Environmental Technology and Innovations. Leiden: CRC Press, 2017 - (Kočí, V.; Ruml, T.; Dao, P.; Duc, P.), s. 21-27. ISBN 978-1-138-02996-5.
[International Conference on Environmental Technology and Innovations /1./. Ho Chi Minh City (VN), 23.11.2016-25.11.2016]
Grant CEP: GA MŠMT LO1302; GA MŠMT(CZ) LO1304
Institucionální podpora: RVO:61388963
Klíčová slova: HIV * inhibitor * screening * assembly * uncoating
Obor OECD: Biochemistry and molecular biology

Nature provides immense variety of biologically active compounds. The beneficial effects of biological materials for humans are well recognized in traditional medicine. However, most of them are still poorly characterized despite immense effort of numerous laboratories. This is in some extent caused also by a quest to identify single compound rather than focusing on synergistic effect of more compounds present in the material either of plant or animal origin. A need for new inhibitors preferentially of natural origin is well justified also for viruses. This is especially true for those viruses that copy genomic material with a low fidelity. This results in continuous emergence of high numbers of drug-resistant viral mutants including those of HIV. In the search for compounds potentially inhibiting HIV, we have recently developed a fluorescent high-throughput assay for screening the inhibitors of assembly of the virus (named FAITH-Fast Assembly Inhibitor Test for HIV) Hadravova et al. (2015). The method was validated using previously reported assembly inhibitors and was shown to be very sensitive and reliable as it provides minimum of false results, as confirmed by electron microscopy. We summarize here the principle and basic protocol of FAITH and we present its application for screening of inhibitors acting in two possible modes. Both of them are essential for complete retrovirus life cycle and thus their inhibition might block virus infectivity, by inhibiting either the assembly of immature particle or disassembly of mature virus core. Based on recently published detailed HIV-1 structure, we designed in silico several compounds presumptively binding to selected interaction interfaces within hexagonal lattice of virus particle. Interestingly, some of the compounds accelerated the assembly, instead of blocking it. This suggests that these compounds bind with a high affinity without preventing the particle formation and thus might block the disassembly of the particle rather than its assembly. As the disassembly is another key step in the virus life cycle, we are currently optimizing the method for screening of the compounds inhibiting the disassembly of the virus core. The method is now being adapted also for other viral species. Both the assembly and disassembly screening methods are applicable also for a non-targeted empirical screening of inhibiting activities of compounds present in biological extracts, preferentially those used in traditional medicine against viral infections. Extremely interesting would be extracts with combined inhibitory activity against HIV and microbial pathogens. These would target not only the primary cause of AIDS but also the microorganisms causing opportunistic infections in AIDS patients.
Trvalý link: http://hdl.handle.net/11104/0297740