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In Vitro Virucidal Activity of Echinaforce®, an Echinacea Purpurea

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In Vitro Virucidal Activity of Echinaforce®, an Echinacea Purpurea Signer et al. Virology Journal (2020) 17:136 https://doi.org/10.1186/s12985-020-01401-2 RESEARCH Open Access In vitro virucidal activity of Echinaforce®, an Echinacea purpurea preparation, against coronaviruses, including common cold coronavirus 229E and SARS-CoV-2 Johanna Signer1†, Hulda R. Jonsdottir1*† , Werner C. Albrich2, Marc Strasser1, Roland Züst1, Sarah Ryter1, Rahel Ackermann-Gäumann1, Nicole Lenz1, Denise Siegrist1, Andreas Suter3, Roland Schoop3 and Olivier B. Engler1* Abstract Background: Coronaviruses (CoVs) were long thought to only cause mild respiratory and gastrointestinal symptoms in humans but outbreaks of Middle East Respiratory Syndrome (MERS)-CoV, Severe Acute Respiratory Syndrome (SARS)-CoV-1, and the recently identified SARS-CoV-2 have cemented their zoonotic potential and their capacity to cause serious morbidity and mortality, with case fatality rates ranging from 4 to 35%. Currently, no specific prophylaxis or treatment is available for CoV infections. Therefore we investigated the virucidal and antiviral potential of Echinacea purpurea (Echinaforce®) against human coronavirus (HCoV) 229E, highly pathogenic MERS- and SARS-CoVs, as well as the newly identified SARS-CoV-2, in vitro. Methods: To evaluate the antiviral potential of the extract, we pre-treated virus particles and cells and evaluated remaining infectivity by limited dilution. Furthermore, we exposed cells to the extract after infection to further evaluate its potential as a prophylaxis and treatment against coronaviruses. We also determined the protective effect of Echinaforce® in re-constituted nasal epithelium. Results: In the current study, we found that HCoV-229E was irreversibly inactivated when exposed to Echinaforce® at 3.2 μg/ml IC50. Pre-treatment of cell lines, however, did not inhibit infection with HCoV-229E and post-infection treatment had only a marginal effect on virus propagation at 50 μg/ml. However, we did observe a protective effect in an organotypic respiratory cell culture system by exposing pre-treated respiratory epithelium to droplets of HCoV-229E, imitating a natural infection. The observed virucidal activity of Echinaforce® was not restricted to common cold coronaviruses, as both SARS-CoV-1 and MERS-CoVs were inactivated at comparable concentrations. Finally, the causative agent of COVID-19, SARS-CoV-2 was also inactivated upon treatment with 50μg/ml Echinaforce®. (Continued on next page) * Correspondence: [email protected]; [email protected] †Johanna Signer and Hulda R. Jonsdottir contributed equally to this work. 1SPIEZ LABORATORY, Austrasse, 3700 Spiez, Switzerland Full list of author information is available at the end of the article © The Author(s). 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. Signer et al. Virology Journal (2020) 17:136 Page 2 of 11 (Continued from previous page) Conclusions: These results show that Echinaforce® is virucidal against HCoV-229E, upon direct contact and in an organotypic cell culture model. Furthermore, MERS-CoV and both SARS-CoV-1 and SARS-CoV-2 were inactivated at similar concentrations of the extract. Therefore we hypothesize that Echinacea purpurea preparations, such as Echinaforce®, could be effective as prophylactic treatment for all CoVs due to their structural similarities. Keywords: Antivirals, Echinacea, Coronavirus, Prevention, HCoV-229E, Common cold, SARS-CoV-1, MERS-CoV, SARS- CoV-2 Background Echinacea plants have traditionally been used in North Coronaviruses (CoVs) are enveloped, positive-sense, America for the prevention and treatment of cold and single-stranded RNA viruses with a large genome, typic- flu symptoms and are now one of the most widely used ally 26–32 kb in length. They belong to the family Coro- medical plants in both North America and Europe [13]. naviridae and are capable of infecting a wide variety of Several different products are on the market, not only hosts [1]. The CoVs capable of causing disease in varying in the Echinacea species and the parts of the humans (HCoVs) were traditionally thought to cause plant used but also in manufacturing procedures, which, only mild gastrointestinal and respiratory tract symp- unfortunately, results in a large variability in quality and toms. Currently, seven HCoVs have been identified. Four activity [14, 15]. Echinaforce® is a standardized prepar- of those, HCoV-229E, HCoV-OC43, HCoV-NL63 and ation extracted from freshly harvested Echinacae pur- HCoV-HKU1, are non-zoonotic and cause worldwide purea plants with a 65% alcoholic solution. outbreaks of upper respiratory tract infections (URTI) Echinaforce® as prevention and treatment of respira- predominantly in the winter period [2]. These com- tory tract infections has been investigated in both pre- monly circulating viruses have been thought to be re- clinical and clinical studies and its beneficial effects sponsible for 10–15% of all URTIs in humans. They documented [16–19]. Specific mechanism of action is replicate in the nasopharynx and generally cause mild, not fully understood but in vitro studies indicate that self-limited URTIs with short incubation periods, al- Echinaforce® inhibits enveloped respiratory viruses in- though lower tract respiratory infections and pneumonia cluding influenza A and B, respiratory syncytial virus have occasionally been described [3–6]. Until the emer- (RSV) or parainfluenza virus, through direct interaction gence of Severe Acute Respiratory Syndrome (SARS)- with whole virions and viral envelope proteins [20, 21]. CoV-1 in 2002, HCoVs were thought to mainly be In general, intracellular activity of Echinacea has been responsible for the common cold. However, the more observed for some viruses (e.g. influenza and herpes sim- virulent coronaviruses, Middle East respiratory syn- plex virus) but not others (e.g. RSV), and only at higher drome (MERS)-CoV and SARS-CoV-1 have animal res- concentrations than required for extracellular inactiva- ervoirs with proposed origins in bats [7] and can cause tion. Furthermore, Echinacea has been shown to severe pneumonias with longer incubation periods and interfere with virus mediated cytokine release [22, 23] often fatal outcome [8]. SARS-CoV-1 was introduced and since typical symptoms of the common cold, i.e. into the human species in 2002 causing a worldwide sneezing, coughing and runny nose, are the results of pandemic, culminating in 8422 infections and 916 deaths the stimulation of pro-inflammatory cytokines, the re- [9]. MERS-CoV is endemic in dromedary camels and duction of cytokine release might help to ease such leads to lower respiratory tract infections in humans with symptoms. In a randomized, double-blind, multi-center, a current case-fatality rate of 35.5% [10]. As of late 2019, a non-inferiority clinical trial Echinaforce® was demon- pneumonia outbreak caused by a novel CoV, designated strated to be non-inferior to Oseltamivir in patients with SARS-CoV-2, supposedly originating from a live seafood influenza-like illness, i.e. involvement of the lower re- market in Wuhan, China, has resulted in a global pan- spiratory tract (cough) and systemic symptoms (e.g. demic with almost 25 million infections and over 800.000 headache, myalgia, fever), and confirmed influenza deaths (WHO situation report, August 31st 2020 and infection with a non-significant trend towards lower in- [11]). To date, there is a lack of established and clinically cidence of complications with Echinaforce Hot Drink® tested antiviral compounds against coronaviruses in gen- compared to Oseltamivir [17]. eral and, more distressingly, the zoonotic betacorona- The antiviral activity of Echinacea has been investi- viruses [12]. Given their increasing incidence and burden, gated in vitro for most of the respiratory viruses finding an inexpensive, accessible and effective treatment associated with common colds and flu, but as of yet, not for HCoVs is of utmost importance. for coronaviruses. Since HCoV-229E is a typical Signer et al. Virology Journal (2020) 17:136 Page 3 of 11 representative of a coronavirus strain causing a seasonal Table 1 Pharmacologically active substances in Echinaforce common cold, we used this virus strain to investigate (batch: 1023117). Data is presented as the mean of four the general antiviral activity of Echinaforce® against cor- independent determinations onaviruses, thereby closing the knowledge gap on the Compound Concentration (μg/ml) antiviral effects of Echinacea purpurea on typical Caffeic acid 0±0 common cold viruses. Furthermore, we expanded
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