Moxidectin and Ivermectin Inhibit SARS-CoV-2 Replication in Vero E6 Cells but Not in Human Primary Bronchial Epithelial Cells
Nilima Dinesh Kumar, Bram M Ter Ellen, Ellen M Bouma, Berit Troost, Denise P I Van De Pol, Heidi H Van Der Ende-Metselaar, Djoke Van Gosliga, Leonie Apperloo, Orestes A Carpaij, Maarten Van Den Berge, Martijn C Nawijn, Ymkje Stienstra, Izabela A Rodenhuis-Zybert, Nilima Dinesh Kumar, Bram M Ter, Jolanda M Smit
Antiviral therapies are urgently needed to treat and limit the development of severe COVID-19 disease. Ivermectin, a broad-spectrum anti-parasitic agent, has been shown to have anti-SARS-CoV-2 activity in Vero cells at a concentration of 5 mM. These limited in vitro results triggered the investigation of ivermectin as a treatment option to alleviate COVID-19 disease. However, in April 2021, the World Health Organization stated the following: "The current evidence on the use of ivermectin to treat COVID-19 patients is inconclusive." It is speculated that the in vivo concentration of ivermectin is too low to exert a strong antiviral effect. Here, we performed a head-to-head comparison of the antiviral activity of ivermectin and the structurally related, but metabolically more stable moxidectin in multiple in vitro models of SARS-CoV-2 infection, including physiologically relevant human respiratory epithelial cells. Both moxidectin and ivermectin exhibited antiviral activity in Vero E6 cells. Subsequent experiments revealed that these compounds predominantly act on the steps following virus cell entry. Surprisingly, however, in human-airway-derived cell models, both moxidectin and ivermectin failed to inhibit SARS-CoV-2 infection, even at concentrations of 10 mM. These disappointing results call for a word of caution in the interpretation of anti-SARS-CoV-2 activity of drugs solely based on their activity in Vero cells. Altogether, these findings suggest that even using a high-dose regimen of ivermectin, or switching to another drug in the same class, is unlikely to be useful for treatment of SARS-CoV-2 in humans. KEYWORDS moxidectin, ivermectin, antiviral, SARS-CoV-2, ALI, in vitro W ithin less than 1.5 years, the pandemic SARS coronavirus 2 (SARS-CoV-2) has infected over 153 million individuals and resulted in over 3.2 million deaths worldwide (1-3). The social and economic burden of this still-ongoing pandemic is staggering, and, besides vaccine development, it is of utmost importance to develop therapeutic interventions to reduce disease symptoms. To date, multiple compounds have been shown to exert SARS-CoV-2 antiviral activity in vitro and several compounds have reached clinical trials (4, 5). Remdesivir and hydroxychloroquine were thought to be effective early in the pandemic, but after a careful evaluation in an interim solidarity trial, the WHO released a conditional yet strong recommendation against the usage of
% cytotoxicity ¼ ðcompound-treated LDH activity 2 spontaneous LDH activityÞ ðmaximum LDH activity 2 spontaneous LDH activityÞ Â 100 Live/dead staining and flow cytometry. PBECs cultured under ALI conditions were treated with 10 mM moxidectin, ivermectin, or an equivalent volume of EtOH at the basolateral side for 48 h at 37°C. Subsequently, cells were harvested by trypsinization and stained with fixable viability dye eFluor780 (Thermo Fisher Scientific) for 20 min at 4°C. Next, cells were washed with fluorescence-activated cell sorter (FACS) buffer (1X phosphate-buffered saline, 2% FBS, 1% EDTA), centrifuged, and fixed with 4% paraformaldehyde for 10 min at 4°C. After fixation, cells were washed, centrifuged, and resuspended in FACS buffer. Flow cytometry analyses were performed using the LSR-2 flow cytometer (BD Biosciences, San Jose, CA, USA) and data was further analyzed using Kaluza analysis software, version 2.1 (Beckman Coulter, Fullerton, CA, USA). Antiviral assays in Vero E6 and Calu-3. Vero E6 cells were seeded at a density of 1.3 Â 10 5 cells/well in 12-well plates. The next day, the medium was replaced with 0.25 ml of DMEM (2% FBS) containing the virus inoculum (MOI 1), in the presence of either increasing concentrations of compounds or the equivalent volume of EtOH. Following 2 h adsorption at 37°C, the virus inoculum was removed, after which the cells were washed twice and fresh DMEM (10% FBS) containing either the compounds or EtOH was added. At 8 hpi, cell..
References
Ahmed, Karim, Ross, Hossain, Clemens et al., A five-day course of ivermectin for the treatment of COVID-19 may reduce the duration of illness, Int J Infect Dis,
doi:10.1016/j.ijid.2020.11.191Bmt, Kumar, Bouma, Troost, Van De Pol et al., Resveratrol and pterostilbene inhibit SARS-CoV-2 replication in air liquid interface cultured human primary bronchial epithelial cells, Viruses,
doi:10.3390/v13071335Bray, Rayner, Noel, Jans, Wagstaff, Ivermectin and COVID-19: a report in antiviral research, widespread interest, an FDA warning, two letters to the editor and the authors' responses, Antiviral Res,
doi:10.1016/j.antiviral.2020.104805Cao, Coyle, Xiong, Wang, Heflich et al., Invited review: human air-liquid-interface organotypic tissue models derived from primary tracheobronchial epithelial cells-overview and perspectives, Vitro Cell Dev Biol Anim,
doi:10.1007/s11626-020-00517-7Chaccour, Casellas, Blanco-Di Matteo, Pineda, Fernandez-Montero et al., The effect of early treatment with ivermectin on viral load, symptoms and humoral response in patients with non-severe COVID-19: A pilot, double-blind, placebo-controlled, randomized clinical trial, EClinicalMedicine,
doi:10.1016/j.eclinm.2020.100720Chowdhury, Shahbaz, Karim, A Randomized Trial of Ivermectin-Doxycycline and Hydroxychloroquine-Azithromycin therapy on COVID-19 patients
Chu, Chan, Yuen, Shuai, Yuan et al., Comparative tropism, replication kinetics, and cell damage profiling of SARS-CoV-2 and SARS-CoV with implications for clinical manifestations, transmissibility, and laboratory studies of COVID-19: an observational study, Lancet Microbe,
doi:10.1016/S2666-5247(20)30004-5De Melo, Benincasa, Cruz, Maricato, Porcionatto, 3D culture models to study SARS-CoV-2 infectivity and antiviral candidates: From spheroids to bioprinting, Biomed J,
doi:10.1016/j.bj.2020.11.009Gonzalez Canga, Prieto, Diez Liebana, Martinez, Vega et al., The pharmacokinetics and metabolism of ivermectin in domestic animal species, Vet J,
doi:10.1016/j.tvjl.2007.07.011Gonzalez, Gámez, Enciso, Maldonado, Palacios et al., Efficacy and safety of ivermectin and hydroxychloroquine in patients with severe COVID-19. A randomized controlled trial,
doi:10.1101/2021.02.18.21252037Gotz, Magar, Dornfeld, Giese, Pohlmann et al., Influenza A viruses escape from MxA restriction at the expense of efficient nuclear vRNP import, Sci Rep,
doi:10.1038/srep25428Hashim, Maulood, Rasheed, Fatak, Kabah et al., Controlled randomized clinical trial on using ivermectin with doxycycline for treating COVID-19 patients in Baghdad,
doi:10.1101/2020.10.26.20219345Heijink, Kies, Kauffman, Postma, Van Oosterhout et al., Down-regulation of E-cadherin in human bronchial epithelial cells leads to epidermal growth factor receptor-dependent Th2 cell-promoting activity, J Immunol,
doi:10.4049/jimmunol.178.12.7678Heijink, Postma, Noordhoek, Broekema, Kapus, House dust mite-promoted epithelial-to-mesenchymal transition in human bronchial epithelium, Am J Respir Cell Mol Biol,
doi:10.1165/rcmb.2008-0449OCJia, Look, Shi, Hickey, Pewe et al., ACE2 receptor expression and severe acute respiratory syndrome coronavirus infection depend on differentiation of human airway epithelia, J Virol,
doi:10.1128/JVI.79.23.14614-14621.2005Jonsdottir, Dijkman, Coronaviruses and the human airway: a universal system for virus-host interaction studies, Virol J,
doi:10.1186/s12985-016-0479-5Ketkar, Yang, Wormser, Wang, Lack of efficacy of ivermectin for prevention of a lethal Zika virus infection in a murine system, Diagn Microbiol Infect Dis,
doi:10.1016/j.diagmicrobio.2019.03.012Lam, Bordin, Waman, Scholes, Ashford et al., SARS-CoV-2 spike protein predicted to form complexes with host receptor protein orthologues from a broad range of mammals, Sci Rep,
doi:10.1038/s41598-020-71936-5Leist, Schafer, Martinez, Cell and animal models of SARS-CoV-2 pathogenesis and immunity, Dis Model Mech,
doi:10.1242/dmm.046581Lopez-Medina, Lopez, Hurtado, Davalos, Ramirez et al., Effect of ivermectin on time to resolution of symptoms among adults with mild COVID-19: a randomized clinical trial, JAMA,
doi:10.1001/jama.2021.3071Lundberg, Pinkham, Baer, Amaya, Narayanan et al., Nuclear import and export inhibitors alter capsid protein distribution in mammalian cells and reduce Venezuelan Equine Encephalitis Virus replication, Antiviral Res,
doi:10.1016/j.antiviral.2013.10.004Makhani, Khatib, Corbeil, Kariyawasam, Raheel et al., 2018 in review: five hot topics in tropical medicine, Trop Dis Travel Med Vaccines,
doi:10.1186/s40794-019-0082-zMastrangelo, Pezzullo, Burghgraeve, Kaptein, Pastorino et al., Ivermectin is a potent inhibitor of flavivirus replication specifically targeting NS3 helicase activity: new prospects for an old drug, J Antimicrob Chemother,
doi:10.1093/jac/dks147Matsuyama, Nao, Shirato, Kawase, Saito et al., Enhanced isolation of SARS-CoV-2 by TMPRSS2-expressing cells, Proc Natl Acad Sci U S A,
doi:10.1073/pnas.2002589117Mcaloose, Laverack, Wang, Killian, Caserta et al., From people to Panthera: natural SARS-CoV-2 infection in tigers and lions at the Bronx Zoo, mBio,
doi:10.1128/mBio.02220-20Mohan, Tiwari, Suri, Mittal, Patel et al., Ivermectin in mild and moderate COVID-19 (RIVET-COV): a randomized, placebo-controlled trial, Res Square Preprint,
doi:10.21203/rs.3.rs-191648/v1Momekov, Momekova, Ivermectin as a potential COVID-19 treatment from the pharmacokinetic point of view: antiviral levels are not likely attainable with known dosing regimens, Biotechnol Biotechnol Equip,
doi:10.1080/13102818.2020.1775118Niaee, Gheibi, Namdar, Allami, Zolghadr et al., Ivermectin as an adjunct treatment for hospitalized adult COVID-19 patients: a randomized multi-center clinical trial, Res Square Preprint,
doi:10.21203/rs.3.rs-109670/v1Nolan, Lok, Macrocyclic lactones in the treatment and control of parasitism in small companion animals, Curr Pharm Biotechnol,
doi:10.2174/138920112800399167Ogando, Dalebout, Zevenhoven-Dobbe, Limpens, Van Der Meer et al., SARS-coronavirus-2 replication in Vero E6 cells: replication kinetics, rapid adaptation and cytopathology, J Gen Virol,
doi:10.1099/jgv.0.001453Okumus, Demirtürk, Çetinkaya, Güner, Avcı et al., Evaluation of the effectiveness and safety of adding ivermectin to treatment in severe COVID-19 patients, Res Square Preprint,
doi:10.21203/rs.3.rs-224203/v1Opoku, Bakajika, Kanza, Howard, Mambandu et al., Single dose moxidectin versus ivermectin for Onchocerca volvulus infection in Ghana, Liberia, and the Democratic Republic of the Congo: a randomized, controlled, doubleblind phase 3 trial, Lancet,
doi:10.1016/S0140-6736(17)32844-1Pan, Peto, Henao-Restrepo, Preziosi, Sathiyamoorthy et al., Repurposed antiviral drugs for COVID-19 -interim WHO solidarity trial results, N Engl J Med,
doi:10.1056/NEJMoa2023184Patterson, Elia, Grassi, Giordano, Desario et al., Evidence of exposure to SARS-CoV-2 in cats and dogs from households in Italy, Nat Commun,
doi:10.1038/s41467-020-20097-0Pena-Silva, Duffull, Steer, Jaramillo-Rincon, Gwee et al., Pharmacokinetic considerations on the repurposing of ivermectin for treatment of COVID-19, Br J Clin Pharmacol,
doi:10.1111/bcp.14476Podder, Chowdhury, Sina, Haque, Outcome of ivermectin treated mild to moderate COVID-19 cases: a single-center, openlabel, randomised controlled study, IMC J Medical Science
Prichard, Geary, Perspectives on the utility of moxidectin for the control of parasitic nematodes in the face of developing anthelmintic resistance, Int J Parasitol Drugs Drug Resist,
doi:10.1016/j.ijpddr.2019.06.002Rajter, Sherman, Fatteh, Vogel, Sacks et al., Use of ivermectin is associated with lower mortality in hospitalized patients with coronavirus disease 2019: the ivermectin in COVID nineteen Study, Chest,
doi:10.1016/j.chest.2020.10.009Rochwerg, Agarwal, Siemieniuk, Agoritsas, Lamontagne et al., A living WHO guideline on drugs for COVID-19, BMJ,
doi:10.1136/bmj.m3379Roy, Pattadar, Raj, Agarwal, Biswas et al., Ivermectin as a potential treatment for mild to moderate COVID-19 -A double blind randomized placebo-controlled trial,
doi:10.1101/2021.01.05.21249310Schmith, Zhou, Lohmer, The approved dose of ivermectin alone is not the ideal dose for the treatment of COVID-19, Clin Pharmacol Ther,
doi:10.1002/cpt.1889Sharun, Tiwari, Natesan, Dhama, SARS-CoV-2 infection in farmed minks, associated zoonotic concerns, and importance of the One Health approach during the ongoing COVID-19 pandemic, Vet Q,
doi:10.1080/01652176.2020.1867776:1-14Shi, Wen, Zhong, Yang, Wang et al., Susceptibility of ferrets, cats, dogs, and other domesticated animals to SARS-coronavirus 2, Science,
doi:10.1126/science.abb7015Sims, Burkett, Yount, Pickles, SARS-CoV replication and pathogenesis in an in vitro model of the human conducting airway epithelium, Virus Res,
doi:10.1016/j.virusres.2007.03.013Sohrabi, Alsafi, Neill, Khan, Kerwan et al., World Health Organization declares global emergency: a review of the 2019 novel coronavirus (COVID-19), Int J Surg,
doi:10.1016/j.ijsu.2020.02.034Soto-Becerra, Culquichicón, Hurtado-Roca, Araujo-Castillo, Real-world effectiveness of hydroxychloroquine, azithromycin, and ivermectin among hospitalized COVID-19 patients: results of a target trial emulation using observational data from a nationwide healthcare system in Peru,
doi:10.1101/2020.10.06.20208066Tay, Fraser, Chan, Moreland, Rathore et al., Nuclear localization of dengue virus (DENV) 1-4 nonstructural protein 5; protection against all 4 DENV serotypes by the inhibitor Ivermectin, Antiviral Res,
doi:10.1016/j.antiviral.2013.06.002Varghese, Kaukinen, Glasker, Bespalov, Hanski et al., Discovery of berberine, abamectin and ivermectin as antivirals against chikungunya and other alphaviruses, Antiviral Res,
doi:10.1016/j.antiviral.2015.12.012Vieira Braga, Kar, Berg, Carpaij, Polanski et al., A cellular census of human lungs identifies novel cell states in health and in asthma, Nat Med,
doi:10.1038/s41591-019-0468-5Wagstaff, Rawlinson, Hearps, Jans, An AlphaScreen (R)-based assay for high-throughput screening for specific inhibitors of nuclear import, J Biomol Screen,
doi:10.1177/1087057110390360Wagstaff, Sivakumaran, Heaton, Harrich, Jans, Ivermectin is a specific inhibitor of importin alpha/beta-mediated nuclear import able to inhibit replication of HIV-1 and dengue virus, Biochem J,
doi:10.1042/BJ20120150Who, Coronavirus disease (COVID-19) pandemic
Who, Therapeutics and COVID-19: living guidelines
Yamasmith, Avirutnan, Mairiang, Tanrumluk, Suputtamongkol et al., Efficacy and safety of ivermectin against dengue infection: a phase III, randomized, double-blind, placebo-controlled trial
Zhang, Xie, Hashimoto, Current status of potential therapeutic candidates for the COVID-19 crisis, Brain Behav Immun,
doi:10.1016/j.bbi.2020.04.046Zhou, Yang, Wang, Hu, Zhang et al., A pneumonia outbreak associated with a new coronavirus of probable bat origin, Nature,
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'Accessed 24 February 2021.'},
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