PMC:7755033 / 2086-14590 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/7755033","sourcedb":"PMC","sourceid":"7755033","source_url":"https://www.ncbi.nlm.nih.gov/pmc/7755033","text":"1. Introduction\nAs per the World Health Organization (WHO), viral diseases are on the rise posing serious public health concerns. Over the last twenty years, numerous viral epidemics such as the Severe Acute Respiratory Syndrome (SARS) in 2002–2003, H1N1 influenza in 2009 and Middle East Respiratory Syndrome (MERS) in 2012 have been reported. Wuhan, the sprawling capital of Central China’s Hubei province very recently witnessed ‘pneumonia of unknown etiology’ that was first reported to the WHO Country Office in China on December 31, 2019. The causal agent coronavirus belonging to the family coronaviridae are enveloped viruses which are divided into four genera (α, β, γ, and δ). The SARS-CoV-2 virus (formerly designated as COVID-19) belonging to the class of β-coronaviruses is supposed to have evolved from bats and thereafter, spread like wildfire worldwide. A total of 43,825,003 confirmed cases of COVID-19 and 1,165,289 deaths have been registered round the globe, whereas in India the confirmed cases has reached to 7,946,429 with 119,535 deaths as of October 27, 2020 (https://www.worldometers.info/coronavirus/country/india/).\nCoronaviruses are a large family of single stranded RNA (ssRNA) viruses ranging from 60 to 140 nm in diameter. Under the electron microscope, they appear to possess spike like projections present on the surface giving it a crown like appearance, hence named ‘coronaviruses’ (Richman et al., 2016). SARS-CoV-2 virus spreads faster than its two predecessors (SARS-CoV and MERS-CoV), but has a lower fatality rate though it is highly contagious. Coronavirus possesses four main structural proteins : Spike (S) protein, membrane (M) protein, envelope (E) protein and nucleocapsid (N) protein (Bosch et al., 2003). Published literature has tracked the onset of symptoms which include a pandemic of cases with pneumonia, unexplained lower respiratory tract infections and multi-organ dysfunction. On the other hand, many reported cases are asymptomatic. SARS-CoV-2 has become one of the major disease pathogens in emerging outbreaks.\nIntense co-operative worldwide efforts and aggressive isolation measures have more or less led to a progressive decline in the number of incidents. The political and health authorities are making an extraordinary effort to halt the shock wave which is seriously challenging the health system globally. Currently, therapeutic options are limited and preventive attempts to minimize social transmission are our best defence weapons. Researchers are working diligently to find novel and/or repurposed therapeutic options to prevent and treat this global pandemic. In this process, they have laid down three strategies , the first includes testing of existing broad-spectrum antiviral drugs (Chan et al., 2013), the second strategy is to use molecular dynamics tools to screen for natural molecules effective as therapeutic and preventive drugs (de Wilde et al., 2014; Dyalla et al., 2014), and third strategy includes the direct utilization of genomic information and pathological symptoms of different mutated coronaviruses to develop new targeted drugs from scratch (Zumla et al., 2016).\nTill date, research has identified more than 30 therapeutic agents, including natural products, allopathic drugs and traditional Chinese medicines which could potentially be effective against COVID-19. For initial treatment of COVID-19, the People's Republic of China National Health Commission (NHC) has included antiviral agents including interferon α (IFN-α), lopinavir/ritonavir, chloroquine phosphate, ribavirin and arbidol in the revised edition of the guidelines for the prevention, diagnosis and treatment of novel coronavirus-induced pneumonia. Combinations of protease inhibitor(s) lopinavir/ritonavir for the treatment of COVID-19 infected patients have been evaluated which have been used previously to treat HIV patients (Liu et al., 2020). In several animal models, including non-human primates (Fouchier et al., 2003; Kuiken et al., 2003; McAuliffe et al., 2004; Rowe et al., 2004), ferrets (Martina et al., 2003), mice (Glass et al., 2004; Hogan et al., 2004) and Syrians hamsters (Roberts et al., 2005), experimental infections with SARS-CoV have been found to cause severe respiratory tract infections.\nSeveral antiviral therapies have been identified for human pathogenic CoVs including synthetic inhibitors of neuraminidase, nucleoside analogs, remdesivir, tenofovir disoproxil (TDF), umifenovir (arbidol) and lamivudine (3TC) (Li et al., 2005). Studies involving identification of cellular receptors that facilitate the binding and entry of human associated coronaviruses have been done. Angiotensin I-converting enzyme 2 (ACE2) is a membrane bound aminopeptidase expressed in renal, cardiovascular, vascular and testicular tissue, as well as the small intestine (Donoghue et al., 2000; Hamming et al., 2004; Harmer et al., 2002). It has been shown to be a co-receptor for viral entry of SARS-CoV-2 with increasing evidence that it has a significant role in SARS-CoV-2 pathogenesis. Therefore, targeting the human ACE2 receptor might block the entry and the subsequent pathophysiology of the virus and is one of the premises of the present study (Zhou et al., 2020). SARS-CoV-2 entry into host cells is mediated by transmembrane spike(S) like glycoprotein that causes neutralization of antibodies, facilitates the projection of homotrimers from the viral surface, binds to human ACE2 receptor, and finally mediates membrane fusion and transport (Tortorici \u0026 Veesler, 2019). Spike (S) protein initially cleaves itself into two functional subunits (S1 and S2) which remain non-covalently bound to each other in a prefusion conformation. The S1 subunit is responsible for binding to ACE2, whereas S2 subunit has role in viral and host cell membrane fusion (Belouzard et al., 2009; Bosch et al., 2003; Burkard et al., 2014; Kirchdoerfer et al., 2016).\nSARS-CoV-2 also encodes a non-structural replicase polyprotein that is processed by viral proteases viz. the primary viral proteinase (3CL-pro) and the papain-like protease (PL-pro). 3CL-pro (Nsp5) regulates viral replication and cleavage of Nsp4 to Nsp16 (Ziebuhr et al., 2000). This main protease is another attractive therapeutic target (Anand et al., 2003; Liu et al., 2020). PL-pro, another crucial non-structural protein, is a virally encoded cysteine protease which has a role in N-terminal processing of viral polyproteins causing release of Nsp1, Nsp2 and Nsp3 which are essential for viral replication to proceed in a correct manner (Chen et al., 2020; Harcourt et al., 2004). Recently, the structure of two critical non-structural proteins in complex Nsp10/Nsp16 has been reported. The complex is responsible for the modification of genetic material of virus in order to camouflage it with host cell DNA. This permits viral evasion and escape from the immune cells of the host, thereby perpetuating infection. Thus, targeting the Nsp10/Nsp16 complex might help in the effective combat and eradication of COVID-19 infection (https://www.anl.gov/article/new-drug-target-found-for-covid19).\nIn the past decade, hundreds of drugs were withdrawn from the market as a result of their failure to either exhibit the desired biological activity or due to their unexpected toxicity and adverse side effects in clinical trials. These poor outcomes may be attributed due to lack of a planned and systematic approach to rational drug design and discovery leading to losses that place a heavy burden on the poor economic sections of the society in particular and the third world countries in general. In the post genomic era, in silico prediction of chemical leads has a significant role in drug discovery, which has proven to be more time and cost effective (Xu et al., 2012). Clinical studies of various drug like properties are very time consuming and expensive. Therefore, computational techniques come is as helpful and handy before going for the expensive in vitro, in vivo studies and clinical trials. Computational techniques can filter and predict the druglikeness, absorption, distribution, metabolism, excretion, and toxicity (ADMET) criteria of a prospective drug candidate (Cheng et al., 2012). In this context, Ayurveda, the Indian traditional system of medicine encompassing a plethora of medicinal plants and their phytoconstituents(s) with proven diverse pharmacological activities, has remained largely unexplored. Thus, Ayurveda offers a staggering array of natural products and prospective therapeutic agents for evaluation against the causal agent of the ongoing global pandemic.\nWithania somnifera (WS), commonly known as ‘Indian Ginseng’ and ‘Ashwagandha’ in Hindi, belongs to family Solanaceae (Figure 1) and holds an important position in Ayurveda owing to its broad therapeutic spectrum and a plethora of pharmacological activities including anti-spasmodic, anti-arthritic, anti-inflammatory, sedative, nerve soothing, hypotensive, antioxidant, immunomodulatory, anti-stress and anti-tumor (Jain et al., 2012).\nFigure 1. (a) Withania somnifera (L.) Dunal Habit (b) Leaves (c) Flower (d) Fruit (e) Stem (Image courtesy naturesalive.wordpress.com, www.nmpb.nic.in, www.flowersofindia.net, Jain et al. (2012), https://stock.adobe.com/in/search?k=ashwagandha). The plant comprises 29 natural secondary metabolites commonly called as ‘withanolides’ extracted from leaves, stems, roots, and flowers which are known for their potent anti-tumor effect (Table 1). Some previous studies have reported the anti-influenza properties of active constituents of WS against H1N1 influenza and antiviral activity of withanone from WS against novel coronavirus (Balakrishnan et al., 2014; Cai et al., 2015; Varshney et al., 2020). However, still, not much is known regarding the antiviral activity of WS phytoconstituents. Therefore, one of the main aims of the present study was to evaluate the antiviral activity of WS phytoconstituents against major target proteins of SARS-CoV-2, the causal agent of the ongoing pandemic.\nTable 1. List of withanolides from Withania somnifera.\nWithaferin A Withanolide A\nPubChem CID: 265237 PubChem CID: 11294368\nMolecular Formula: C28H38O6 Molecular Formula: C28H38O6\nChemical Class: Steroidal Lactone Chemical Class: Steroidal Lactone\nWithanolide B Withanolide D\nPubChem CID: 14236711 PubChem CID: 161671\nMolecular Formula: C28H38O5 Molecular Formula: C28H38O6\nChemical Class: Steroidal Lactone Chemical Class: Steroidal Lactone\nWithanolide E Withanone\nPubChem CID: 301751 PubChem CID: 21679027\nMolecular Formula: C28H38O7 Molecular Formula: C28H38O6\nChemical Class: Steroidal Lactone Chemical Class: Steroidal Lactone\nViscosalactone B Anaferine\nPubChem CID: 57403080 PubChem CID: 443143\nMolecular Formula: C28H40O7 Molecular Formula: C13H24N2O\nChemical Class: Steroidal Lactone Chemical Class: Piperidine Alkaloid\n \nWithasomnine  \nPubChem CID: 442877  \nMolecular Formula: C12H12N2  \nChemical Class: Pyrazole Alkaloid   Withanolides, including withanolides A, B, D, E, anaferine, withaferin A, withasomnine, withanone and viscosalactone B (Table 1) are responsible for various pharmacological activities as detailed above (Jain et al., 2012; Misra et al., 2008). They belong to chemical classes of alkaloids and steroidal lactones.\nAgents that can preferentially bind to ACE2 receptor can be thought of as promising ‘preventive’ agents against SARS-CoV-2 infection. Alternately, agents that bind to main viral proteins can interfere and/or block crucial stages in the viral life cycle such as viral entry, replication, membrane fusion and hence, can function as promising therapeutic agents. The present paper attempts to highlight the preventive/therapeutic potential of withanolides from WS as promising antiviral agents to augment our limited armamentarium of drugs available against SARS-CoV-2 to effectively combat COVID-19.\nAll the standard reference drugs used in the study like losartan, procainamide, cinacalcet, arbidol, hydroxychloroquine, oberadilol and poziotinib are FDA-approved drugs that were chosen for their specific binding to proteins selected for the study viz. human ACE2 receptor, SARS-CoV and SARS-CoV-2 target proteins (Spike S glycoprotein, protease) as referenced in literature (Arya et al., 2020; Liu et al., 2020, Wu et al., 2020). Since there is no available literature on Nsp10/Nsp16 complex, two randomly selected reference drugs were chosen against it in binding 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