mesЭкстремальная биомедицинаExtreme Medicine3033-89643033-8972Centre for Strategic Planning of the Federal Medical and Biological Agency10.47183/mes.2022.043mes-205Research ArticleОРИГИНАЛЬНОЕ ИССЛЕДОВАНИЕORIGINAL RESEARCHОценка цитотоксичности и противовирусной активности смеси лактоферрина, артемизинина и азитромицина в отношении SARS-CoV-2 in vitroAssessment of cytotoxicity and antiviral activity against SARS-CoV-2 of the mixture of lactoferrin, artemisinin, and azithromycin in vitroРябченковаА. А.RyabchenkovaA. A.

Анастасия Андреевна Рябченкова

пр-кт Малый В. О., д. 57, к. 4, литера Ж, помещение 5-Н, г. Санкт-Петербург, 199178

Anastasia A. Ryabchenkova

Prospect Maly V.O., 57, k. 4, litera Zh, k. 5-Н, St. Petersburg, 199178

riabchenkova@service-gene.ruКопатьВ. В.KopatV. V.

Санкт-Петербург

St. Petersburg

ЧиракЕ. Р.ChirakE. R.

Санкт-Петербург

St. Petersburg

ЧиракЕ. Л.ChirakE. L.

Санкт-Петербург

St. Petersburg

ЛеневаИ. А.LenevaI. A.

Москва

Moscow

ГлубоковаЕ. А.GlubokovaE. A.

Москва

Moscow

КарташоваН. П.KartashovaN. P.

Москва

Moscow

КолмаковН. Н.KolmakovN. N.

Санкт-Петербург

St. Petersburg

ДуховлиновИ. В.DukhovlinovI. V.

Санкт-Петербург

St. Petersburg

Общество с ограниченной ответственностью «АТГ Сервис Ген»РоссияATG Service Gen LLCRussian FederationНаучно-исследовательский институт вакцин и сывороток имени И. И. Мечникова Минобрнауки РоссииРоссияMechnikov Research Institute of Vaccines and SeraRussian FederationИнститут экспериментальной медицины Минобрнауки РоссииРоссияInstitute of Experimental MedicineRussian Federation2022281020242444351Copyright © Рябченкова А.А., Копать В.В., Чирак Е.Р., Чирак Е.Л., Ленева И.А., Глубокова Е.А., Карташова Н.П., Колмаков Н.Н., Духовлинов И.В., 20242024Рябченкова А.А., Копать В.В., Чирак Е.Р., Чирак Е.Л., Ленева И.А., Глубокова Е.А., Карташова Н.П., Колмаков Н.Н., Духовлинов И.В.Ryabchenkova A.A., Kopat V.V., Chirak E.R., Chirak E.L., Leneva I.A., Glubokova E.A., Kartashova N.P., Kolmakov N.N., Dukhovlinov I.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.extrememedicine.ru/jour/article/view/205

Лактоферрин, артемизинин и азитромицин обладают широким спектром противовирусного, иммуномодулирующего и противовоспалительного действия. Экспериментально показанное частичное ингибирование ими инфекции, вызванной SARS-CoV-2 in vitro, позволяет заключить, что влияния на проникновение вирионов в клетки, опосредованное каждым из этих веществ в отдельности, недостаточно для полного ингибирования инфекции SARS-CoV-2. Целью работы было оценить in vitro цитотоксичность и противовирусную активность смеси активных действующих веществ лактоферрина, артемизинина и азитромицина в отношении лабораторного штамма SARS-CoV-2. Использовали перевиваемую культуру клеток Vero CCL81 (ATСС) и лабораторный штамм коронавируса SARS-CoV-2 «Дубровка» (идентификационный № GenBank: MW161041.1), выделенный на культуре клеток Vero CCL81 из назофарингеального мазка больного СOVID-19. Определение цитотоксического действия смеси препаратов и изучение противовирусной активности в отношении вируса SARS-CoV-2 оценивали по эффекту цитопатического действия с использованием МТТ (метилтиазолилдифенил- тетразолия бромид). В качестве препарата сравнения использовали гидроксихлорохин. Показано, что при высокой множественности заражения (100 MOI) и низкой (20 MOI) в культуре клеток Vero ССL81 смеси артемизинина, лактоферрина и азитромицина оказывает значимый эффект на вирусную репродукцию SARS-CoV-2, ИК50 (полумаксимальная ингибирующая концентрация) оценивается в обоих случаях как разведение 1 : 2. Полученные результаты позволяют сделать вывод о низкой цитотоксичности изучаемой смеси и о наличии значимого противовирусного действия in vitro.

Lactoferrin, artemisinin, and azithromycin exhibit a broad spectrum of antiviral, immunomodulatory, and anti-inflammatory effects. The experiments show that these drugs partially inhibit the infection caused by SARS-CoV-2 in vitro. This allows us to conclude that the effects on the entry of virions into cells mediated by each of these substances taken separately are insufficient for complete inhibition of the SARS-CoV-2 infection. The study was aimed to perform in vitro assessment of cytotoxicity and antiviral activity against the laboratory SARS-CoV-2 strain of the mixture of active ingredients: lactoferrin, artemisinin, and azithromycin. We used the Vero CCL81 (ATСС) cell line and the Dubrovka laboratory strain of SARS-CoV-2 (GenBank ID: MW161041.1), isolated in the Vero CCL81 cell culture from the nasopharyngeal swab of patient with СOVID-19. Cytotoxic effects and antiviral activity against SARS-CoV-2 of the drug mixture were assessed based on the cytopathic effects using the MTT (methylthiazolyldiphenyl-tetrazolium bromide) assay. Hydroxychloroquine was used as a reference drug. It has been shown that at high (MOI 100) and low (MOI 20) multiplicity of infection used in the Vero CCL 81 cell culture, the mixture of artemisinin, lactoferrin and azithromycin has a significant effect on the SARS-CoV-2 reproduction, and IC50 (half maximal inhibitory concentration) is estimated as the 1 : 2 dilution in both cases. The findings make it possible to conclude that the studied mixture is low toxic and shows significant antiviral effects in vitro.

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The authors declare that there are no conflicts of interest present.