A small in-house library was screened by treating cells with the compounds prepared in DMSO at 10 µM concentration for 48 h. We used boceprevir (Cas # 394730600, Sigma) as a positive control that showed activity against SARS-COV-2 3CLpro [39 (link)]. The experiments were run on three occasions to identify the assay's hit rate, the reproducibility of the assay, and the false negative and false positive hit rates in the assay. At the endpoint of the split-GFP-3CLpro assay, we measure the compound cytotoxicity by applying 20/well of the MTT solution (CellTiter 96® AQueous, Promega) and incubating the cells for 4 h at 37 °C and 5% CO2. The absorbance was measured at 490 nm using a 96-well plate reader.
Boceprevir
Manufactured by Merck Group
Sourced in Canada, United States
Boceprevir is a protease inhibitor used in the treatment of hepatitis C virus infection. It functions by inhibiting the hepatitis C virus NS3/4A protease, an enzyme essential for viral replication. Boceprevir is utilized as part of a combination therapy regimen for the management of chronic hepatitis C.
Automatically generated - may contain errors
Lab products found in correlation
Celltiter 96 aqueous, by Promega (1 mentions)
Glycyrrhizic acid ammonium salt, by Merck Group (1 mentions)
Gc376, by Merck Group (1 mentions)
Tdzd 8, by Merck Group (1 mentions)
Poly 1 c, by InvivoGen (1 mentions)
Alisporivir, by Novartis (1 mentions)
Daclatasvir, by Bristol-Myers Squibb (1 mentions)
Nim811, by Novartis (1 mentions)
Cyclosporine, by Novartis (1 mentions)
4 protocols using boceprevir
1
Screening for SARS-CoV-2 3CLpro Inhibitors
2
Systematic Evaluation of Compounds
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Boceprevir, GC376, TDZD-8, lercanidipine hydrochloride, efonidipine hydrochloride monoethanolate, and glycyrrhizic acid ammonium salt from Glycyrrhiza root were purchased from Sigma. Stocks of all the compounds were made in dimethyl sulfoxide (DMSO) at a concentration of 20 mM and stored frozen at −80°C.
3
Comparing Viperin Induction by IFN Treatments
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It is known that viperin expression is induced after stimulation of type I IFN in HepG2, Huh7 and fibroblasts cells (13 (link)). Here, we compared six different treatments including 3 cell groups that were transfected with expressing plasmids 48 hours before recombinant IFN (Sigma, UK) treatment, one group was treated only by recombinant IFN alpha (1000 IU/mL) as positive control, mock group treated only with lipofectamine 2000, and the last group was a negative control group without any treatment. In addition, we used a newly approved anti-protease drug, Boceprevir (Merck Co. Canada) in 14 nm concentration to confirm our results by its protease blocking activity in all the treated groups 6 hours after IFN induction.
4
Antiviral Compounds Sourcing Protocol
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The HCV NS5A inhibitor daclatasvir (Bristol Myers Squibb), the HCV NS5B polymerase inhibitor sofosbuvir (Gilead), the HCV NS3 protease inhibitors boceprevir (Merck) and telaprevir (Vertex) and the HIV-1 reverse transcriptase inhibitor emtricitabine (Gilead) were all obtained from MedChemexpress (Princeton, NJ 08540, USA). Alisporivir and NIM811 were generously provided by Novartis, whereas cyclosporine A, sanglifehrins A and B were generously provided by Drs. Wilkinson and Gregory. Poly I:C was obtained from InvivoGen (San Diego, CA, USA).
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