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5 protocols using elbasvir

1

High-Throughput ABCB1 Inhibition Assay

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[3H]-digoxin was purchased from Moravek Biochemicals (Brea, CA, USA). Abacavir, atazanavir, etravirine, lopinavir, maraviroc, rilpivirine, ritonavir, saquinavir, tenofovir DF, and zidovudine were obtained from the NIH AIDS Reagent Program. Asunaprevir, daclatasvir, darunavir, dolutegravir, elbasvir, grazoprevir, ledipasvir, sofosbuvir, and velpatasvir were acquired from MedChemExpress LLC (Middlesex County, NJ, USA). The model ABCB1 inhibitor CP100356 [76 (link)], the ATP bioluminescence assay kit, DMSO, Dulbecco’s Modified Eagle Medium, ethanol (EtOH), fetal bovine serum, nonessential amino acid solution, and penicillin–streptomycin solutions were purchased from Sigma-Aldrich (St. Louis, MO, USA). Hanks’ balanced salt solution, William’s medium E containing L-glutamine (WME), and the bicinchoninic acid protein assay kit were obtained from Thermo Fisher Scientific (Waltham, MA, USA). Krebs–Henseleit buffer was prepared as described by de Graaf et al. [31 (link)]. All other reagents were of analytical grade.
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2

Synthesis of Hepatitis C Antivirals

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Glecaprevir, pibrentasvir and ombitasvir were synthesized at AbbVie. Daclatasvir, elbasvir, ledipasvir, and velpatasvir were purchased from MedChem Express (Monmouth Junction, NJ, USA).
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Evaluation of Hepatitis C Antivirals

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Daclatasvir, velpatasvir, and elbasvir were purchased from MedchemExpress; sofosbuvir was purchased from Selleckchem; and pibrentasvir was purchased from MedKoo.
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4

Antiviral Drug Potency Assay

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Daclatasvir, elbasvir, ledipasvir and velpatasvir were purchased from MedchemExpress; pibrentasvir was purchased from Acme Bioscience. RNA was electroporated and cells were seeded as above.
Compounds were serially diluted 1:2 or 1:10 in DMSO and added to the cells. Only DMSO was added in the control samples. Cells were incubated for 72 hours at 37°C before cell culture medium was removed and luciferase assay was performed. Each experiment included three replicates and was performed at least twice. Nonlinear regression on GraphPad Prism 7.00 (Graph-Pad Software, La Jolla, CA) was used to calculate the EC50 values.
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5

Antiviral Drug Screening Protocol

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Acetaminophen (catalog number HY-66005), arbidol hydrochloride (HY-14904A), aspirin (HY-14654), atazanavir sulfate (HY-17367A), baloxavir marboxil (HY-109025), daclatasvir (HY-10466), darunavir (HY-17040), dolutegravir (HY-13238), efavirenz (HY-10572), elbasvir (HY-15789), etravirine (HY-90005), galidesivir (BCX4430, HY-18649A), ganciclovir (HY-13637), glecaprevir (HY-17634), hydroxychloroquine (HY-B1370), indomethacin (HY-14397), ledipasvir (HY-15602), lopinavir (HY-14588), losartan (HY-17512), nafamostat mesylate (HY-B0190A), nitazoxanide (HY-B0217), ombitasvir (HY-13997), oseltamivir (HY-13318), penciclovir (HY-17424), perindopril salt (HY-B0130A), raltegravir (HY-10353), remdesivir (GS-5734, HY-104077), rilpivirine (HY-10574), ritonavir (HY-90001), simeprevir (HY-10241), sofosbuvir (HY-15005), and β-d-N4-hydroxycytidine (HY-125033) were purchased from MedChemExpress (Monmouth Junction, NJ, United States). Amodiaquin dihydrochloride dihydrate (A2799), artesunate (A3731), chloroquine phosphate (PHR1258), mefloquine hydrochloride (PHR1705), peramivir trihydrate (SML2486), ribavirin (R9644), teicoplanin (T0578), and toremifene citrate salt (T7204) were purchased from Sigma-Aldrich (St, Louis, Missouri, United States). Favipiravir (T-705) was provided by the Toyama Chemical Co., Ltd. (Japan). The drug solutions were filtered through a 0.22 μM membrane and were stored at −80°C.
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