Repurposing of clinically approved drugs for treatment of coronavirus disease 2019 in a 2019-novel coronavirus-related coronavirus model

Abstract

Background Medicines for the treatment of 2019-novel coronavirus (2019-nCoV) infections are urgently needed. However, drug screening using live 2019-nCoV requires high-level biosafety facilities, which imposes an obstacle for those institutions without such facilities or 2019-nCoV. This study aims to repurpose the clinically approved drugs for the treatment of coronavirus disease 2019 (COVID-19) in a 2019-nCoV-related coronavirus model. Methods A 2019-nCoV-related pangolin coronavirus GX_P2V/pangolin/2017/Guangxi was described. Whether GX_P2V uses angiotensin-converting enzyme 2 (ACE2) as the cell receptor was investigated by using small interfering RNA (siRNA) -mediated silencing of ACE2. The pangolin coronavirus model was used to identify drug candidates for treating 2019-nCoV infection. Two libraries of 2406 clinically approved drugs were screened for their ability to inhibit cytopathic effects on Vero E6 cells by GX_P2V infection. The anti-viral activities and anti-viral mechanisms of potential drugs were further investigated. Viral yields of RNAs and infectious particles were quantified by quantitative real-time polymerase chain reaction (qRT-PCR) and plaque assay, respectively. Results The spike protein of coronavirus GX_P2V shares 92.2% amino acid identity with that of 2019-nCoV isolate Wuhan-hu-1, and uses ACE2 as the receptor for infection just like 2019-nCoV. Three drugs, including cepharanthine (CEP), selamectin, and mefloquine hydrochloride, exhibited complete inhibition of cytopathic effects in cell culture at 10 mmol/L. CEP demonstrated the most potent inhibition of GX_P2V infection, with a concentration for 50% of maximal effect [EC 50 ] of 0.98 mmol/L. The viral RNA yield in cells treated with 10 mmol/L CEP was 15,393-fold lower than in cells without CEP treatment ([6.48 ± 0.02] Â 10 À4 vs. 1.00 ± 0.12, t = 150.38, P < 0.001) at 72 h post-infection (p.i.). Plaque assays found no production of live viruses in media containing 10 mmol/L CEP at 48 h p.i. Furthermore, we found CEP had potent anti-viral activities against both viral entry (0.46 ± 0.12, vs.1.00 ± 0.37 t = 2.42, P < 0.05) and viral replication ([6.18 ± 0.95] Â 10 À4 vs. 1.00 ± 0.43, t = 3.98, P < 0.05). Conclusions Our pangolin coronavirus GX_P2V is a workable model for 2019-nCoV research. CEP, selamectin, and mefloquine hydrochloride are potential drugs for treating 2019-nCoV infection. Our results strongly suggest that CEP is a wide-spectrum inhibitor of pan-betacoronavirus, and further study of CEP for treatment of 2019-nCoV infection is warranted.Original Article * P < 0.01). Vero cells were transfected with 0.8, 4, and 20 nmol/L ACE2-specific siRNAs or NSC siRNA by using the RNAiMax transfection reagent. At 48 h p.t., the cells were infected with pangolin CoV at an MOI of approximately 10. At 24 h p.i., CoV-infected cells were lysed in lysis buffer, and the RNA levels of ACE2, CoV, and b-actin were determined by qRT-PCR.

COVID-19, Modern Pandemic: A Systematic Review From Front-Line Health Care Providers’ Perspective

“…It can inhibit 2019-nCoVr at a low concentration. Compared to regular infections without drug treatment, the viral RNA yields were significantly lower in vitro when using CEP; thus, our data suggest that CEP can potently inhibit coronavirus infection at viral entry and postentry phases [51]. CEP has low toxicity in animals and has no significant side effects in humans [51].…”

Shell disorder analysis predicts greater resilience of the SARS-CoV-2 (COVID-19) outside the body and in body fluids

“…This suggests that bat CoVs are likely to have an optimal mix of fecal-oral and respiratory transmission potentials that reflect their N PID range of 47-48%, which provides for greater fitness for the spread of the viruses among bats and that SARS-CoV-2 is likely to be of bat origin too given its N PID is 48%. There are highly debatable suggestions that snakes or pangolins [3,37,38] may be an intermediary host of the virus. Even if any of these is later deemed as true, our data imply that the virus had not the chance to evolved much with any intermediary host except bats, unlike the cases of SARS-CoV and MERS-CoV, in which the model does suggest that they had evolved with hosts such as civet cats and camels respectively.…”

In Silico Discovery of Candidate Drugs against Covid-19

“…Among possible interesting drugs for Covid-19 treatment, we found Nimesulide, Fluticasone Propionate, Thiabendazole, Photofrin, Didanosine and Flutamide.While the diagnosis of COVID-19 is based on the amplification of the viral genome in real-time PCR with specific probes, the current treatment of affected individuals is limited to a mixture of a broad-spectrum of antiviral drugs [6]. However, in many cases this pharmacological approach has proven to be totally ineffective.Screening drug studies on pangolin SARS-CoV-2, which is the human-related Coronavirus, demonstrated that three drugs (cepharanthine, selamectin and mefloquine hydrochloride) were effective in inhibiting viral replication, with cepharanthine potently inhibiting coronavirus infection at viral entry and post-entry viral replication [7]. The last drug is an anti-inflammatory and antineoplastic alkaloid approved for leukopenia and it is also proposed to inhibit the human immunodeficiency virus type 1 (HIV) entering in cells by reducing plasma membrane fluidity.…”

“…Screening drug studies on pangolin SARS-CoV-2, which is the human-related Coronavirus, demonstrated that three drugs (cepharanthine, selamectin and mefloquine hydrochloride) were effective in inhibiting viral replication, with cepharanthine potently inhibiting coronavirus infection at viral entry and post-entry viral replication [7]. The last drug is an anti-inflammatory and antineoplastic alkaloid approved for leukopenia and it is also proposed to inhibit the human immunodeficiency virus type 1 (HIV) entering in cells by reducing plasma membrane fluidity.…”