PHYSICOCHEMICAL PROPERTIES AND BIOLOGICAL ACTIVITY OF THE NEW ANTIVIRAL SUBSTANCE
Keywords:Antiviral pharmaceutical substance, Rimantadine, L-histidyl-1-adamantylethylamine, Quality control
Objective: To develop a set of quality control procedures for the promising antiviral pharmaceutical substance L-histidyl-1-adamantylethylamine dihydrochloride monohydrate, a derivative of rimantadine.
Methods: Substances and solvents: synthesized in laboratory L-histidyl-1-adamantylethylamine dihydrochloride monohydrate (H-His-Rim•2HCl•H2O), rimantadine hydrochloride (Rim•HCl), 99%, ethanol 96%, N, N-dimethylformamide (DMF) anhydrous, 99.8% and n-hexane anhydrous, 95%, deionized high-resistance water (18.2 MΩ•cm at 25 °C, Milli-Q system), silver nitrate. Infrared (IR) Spectroscopy–Cary 630 Fourier Transform IR Spectrometer, elemental analysis–elemental composition analyzer CHNS-O EuroEA3000, ultraviolet (UV) spectrometry–Cary-60 spectrophotometer, polarimetry–POL-1/2 polarimeter with an external Peltier module, granulometric analysis by optical microscopy (Altami BIO 2 microscope) and low-angle laser light scattering (LALLS)–Master Sizer 3600, measurement of potential for hydrogen–potentiometer PB-11, Spirotox method–the study of temperature dependences of Spirostomum ambiguum lifetime to characterize the biological activity of the studied compounds.
Results: The substance H-His-Rim•2HCl•H2O is an amorphous yellowish powder, slightly soluble in water, soluble in ethanol, freely soluble in N, N-dimethylformamide, and practically insoluble in n-hexane. A study of the elemental composition has confirmed the authenticity of H-His-Rim•2HCl•H2O. Comparison of the spectral characteristics of H-His-Rim•2HCl•H2O and Rim•HCl by IR spectroscopy and UV spectrometry confirmed the authenticity of the substance. The racemic form of the substance Rim•HCl with an insignificant amount of impurity of the levorotatory enantiomer was proved polarimetrically: α =-0.0126±0.0003 (1% aqueous solution, 20±0.5 °С). The specific optical rotation of 1% aqueous solution H-His-Rim•2HCl•H2O . In 1% ethanol solution -10.32±0.12. Using the method of laser light diffraction for a substance H-His-Rim•2HCl•H2O, the dimensional spectra «fraction of particles, %-d, μm» were characterized, the maximum of which in hexane is in the region of 40–50 μm. Arrhenius’s kinetics on the Spirotox model established statistically significant differences in ligand-receptor interactions, which are characterized by values of observed apparent activation energy °bsEa, kJ/mol: 132.36±1.55 for H-His-Rim•2HCl•H2O and 176.15±0.48 for Rim•HCl.
Conclusion: The developed set of methods for assessment of physical and chemical properties and biological activity of a new antiviral substance H-His-Rim•2HCl•H2O is the basis for establish of regulatory documentation.
Shie JJ, Fang JM. Development of effective anti-influenza drugs: congeners and conjugates-a review. J Biomed Sci 2019;26:84.
Haque SKM, Elzagheid M. Antiviral drugs for influenza viruses. Int J Pharm Pharm Sci 2019;11:1-10.
Hay AJ, Wolstenholme AJ, Skehel JJ, Smith MH. The molecular basis of the specific anti-influenza action of amantadine. EMBO J 1985;4:3021-4.
Pielak RM, Chou JJ. Flu channel drug resistance: a tale of two sites. Protein Cell 2010;1:246-58.
Bright RA, Medina MJ, Xu X, Perez Oronoz G, Wallis TR, Davis XM. Incidence of adamantane resistance among influenza A (H3N2) viruses isolated worldwide from 1994 to 2005: a cause for concern. Lancet 2005;366:1175–81.
Fiore AE, Fry A, Shay D, Gubareva L, Bresee JS, Uyeki TM. Antiviral agents for the treatment and chemoprophylaxis of influenza: recommendations of the advisory committee on immunization practices (ACIP). MMWR Recomm Rep 2011;60:1–24.
Fytas C, Kolocouris A, Fytas G, Zoidis G, Valmas C, Basler CF. Influence of an additional amino group on the potency of aminoadamantanes against influenza virus A. II-Synthesis of spiropiperazines and in vitro activity against influenza a H3N2 virus. Bioorg Chem 2010;38:247-51.
Li F, Ma C, Hu Y, Wang Y, Wang J. Discovery of potent antivirals against amantadine-resistant influenza viruses by targeting the M2-S31N proton channel. ACS Infect Dis 2016;2:726-33.
Kuznetsov NY, Tikhov RM, Godovikov IA, Medvedev MG, Lyssenko KA, Burtseva EI, et al. Stereoselective synthesis of novel adamantane derivatives with high potency against rimantadine-resistant influenza a virus strains. Org Biomol Chem 2017;15:3152-7.
Hu Y, Musharrafieh R, Ma C, Zhang J, Smee DF, DeGrado WF, et al. An M2-V27A channel blocker demonstrates potent in vitro and in vivo antiviral activities against amantadine-sensitive and-resistant influenza viruses. Antiviral Res 2017;140:45-54.
Shibnev VA, Deryabin PG, Garaev TM, Finogenova MP, Botikov AG, Mishin DV. Peptide carbocyclic derivatives as inhibitors of the viroporin function of RNA-containing viruses. Russ J Bioorg Chem 2017;43:517-25.
Shibnev VA, Garaev TM, Finogenova MP, Shevchenko ES, Burtseva EI. New adamantane derivatives can overcome resistance of influenza A(H1N1)pdm2009 and A(H3N2) viruses to «remantadine». B Exp Biol Med 2012;153:200–2.
Shhelkanov MJu, Shibnev VA, Finogenova MP, Fedjakina IT, Garaev TM, Markova NV, et al. The antiviral activity of the adamantane derivatives against the influenza virus a (H1N1) pdm09 model in vivo. Voprosy Virusologii 2014;59:37–40.
Deryabin PG, Garaev TM, Finogenova MP, Odnovorov AI. Assessment of the antiviral activity of 2HCl•H-His-Rim compound compared to the anti-influenza drug arbidol for influenza caused by A/duck/Novosibirsk/56/05 (H5N1) (Influenza a virus, alphainfluenzavirus, orthomyxoviridae). Voprosy Virusologii 2019;64:268-73.
The State Pharmacopoeia of the Russian Federation 14 (XIV) Edition; 2018.
Zauer EA. Modern automatic CHNS/O/X organic compound analyzers. Anal Control 2018;22:6-19.
Uspenskaya EV, Anfimova EV, Syroeshkin AV, Pleteneva TV. Kinetics of pharmaceutical substance solubility in water with different hydrogen isotopes content. Indian J Pharm Sci 2018;80:318-24.
Goncharuk VV, Syroeshkin AV, Zlatskiy IA, Uspenskaya EV, Orekhova AV, Levitskaya OV, et al. Quasichemical description of the cell death kinetics of cellular biosensor spirostomum ambigua for testing the biological activity of aqueous solutions. J Water Chem Technol 2017;39:97-102.
Emshanova SV, Abramovich RA, Potanina OG. Influence of particle shape and size of substances on the quality of finished medicines. Drug Dev Registration 2014;2:50-63.
Nałecz Jawecki G. Spirotox spirostomum ambiguum acute toxicity test-10 y of experience. Environ Toxicol 2004;19:359-64.
Syroeshkin AV, Uspenskaya EV, Pleteneva TV, Morozova MA, Maksimova TV, Koldina AM, et al. Mechanochemical activation of pharmaceutical substances as a factor for modification of their physical, chemical and biological properties. Int J Appl Pharm 2019;11:118-23.