Timur Mansurovich Garaev
1* 
, Artyom Irorevich Odnovorov
2 , Alexander Aleksandrovich Lashkov
3 , Tatiana Vladimirovna Grebennikova
1 , Marina Pavlovna Finogenova
1 , Galina Kadymovna Sadykova
1 , Alexei Gennadievich Prilipov
1 , Tatiana Anatol'evna Timofeeva
1, Sergey Vadimovich Rubinsky
3, Svetlana Nikolaevna Norkina
1, Marina Mikhailovna Zhuravleva
11 Federal State Budgetary Institution «National Research Centre for Epidemiology and Microbiology named after the honorary academician N.F.Gamaleya» of the Ministry of Health of the Russian Federation (N.F.Gamaleya NRCEM), 123098, Moscow, Russian Federation.
2 Peoples Friendship University of Russia (RUDN University), Ministry of Education of the Russian Federation, 117198, Moscow, Russian Federation.
3 FSRC «Crystallography and Photonics» RAS, Leninskiy Prospekt 59, 119333, Moscow, Russia.
Abstract
Purpose: The aminoadamantane derivative of L-histidyl-1-adamantayl ethylamine hydrochloride (HCl*H-His-Rim) has showed a high inhibition level against influenza A virus strains in vitro. The aim of this work is to search and establish evidence of the direct effect of the drug on influenza A virus proton channel M2.
Methods: The compound HCl*H-His-Rim was obtained by classical peptide synthesis methods. Influenza A virus mutants of A/PuertoRico/8/34(H1N1) strain were obtained by reverse genetics methods. The mutant samples of the virus were cultured on chicken embryos with a virus titer in the hemagglutination test. ELISA was carried out on Madin-Darby canine kidney (MDCK) monolayer cells when multiplying the virus 10-4-10-6. The binding stability of HCl*H-His-Rim was compared to those of M2 (S31N) and M2 (S31N_A30T) channels by molecular dynamic (MD) modeling. The calculation was performed taking into account the interaction with the model lipid bilayer (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) in the presence of water molecules in accordance with the three-center model.
Results: It was found that HCl*H-His-Rim is a direct action drug against influenza A. The most likely conformation of drug binding to target protein has been shown. It has been found that the A30T mutation reduces the binding energy of the drug, and the results obtained in vitro have confirmed the data calculated in silico.
Conclusion: The mechanism of action of HCl*H-His-Rim is directly related to the suppression of the function of the proton channel M2 of influenza A virus.