To determine the antiviral activity of niclosamide, Huh-7 cells were infected with DENV-1, DENV-2, DENV-3, DENV-4, or ZIKV at a multiplicity of infection (MOI) of 0.5 and treated with either niclosamide (Sigma-Aldrich, St. Louis, MO, USA) or a solvent control, dimethyl sulfoxide (DMSO; Sigma-Aldrich), at the indicated concentrations. Dose-response curves were obtained with 3-fold serial dilutions of niclosamide ranging from 0.04 μM to 10 μM. At 24 h p.i., live cells and cell culture supernatants were collected and processed for flow activated cell sorter (FACS) analysis and virus titration, respectively.
Niclosamide
Manufactured by Merck Group
Sourced in United States, Germany, China, Spain, Sao Tome and Principe
Niclosamide is a lab equipment product manufactured by Merck Group. It is a chemical compound used for various research and analytical applications. The core function of Niclosamide is to serve as a reference standard or reagent in laboratory settings.
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Lab products found in correlation
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Icrt14, by Merck Group (3 mentions)
Stat3, by Cell Signaling Technology (3 mentions)
Rotenone, by Merck Group (3 mentions)
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Iwp 2, by Merck Group (2 mentions)
Cyclin d1, by Cell Signaling Technology (2 mentions)
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Tyrphostin a9, by Merck Group (2 mentions)
Doxycycline, by Merck Group (2 mentions)
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Sodium chloride (nacl), by Merck Group (2 mentions)
89 protocols using niclosamide
1
Evaluating Niclosamide's Antiviral Efficacy
2
Synergistic Antimicrobial Activity of Colistin and Niclosamide
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MIC of colistin (Sigma, Spain), MIC of niclosamide (Sigma, Spain), and MIC of colistin in presence of different concentrations of niclosamide (between 0.5 and 4 μM) against Col-S and Col-R references and clinical A. baumannii and K. pneumoniae strains were determined in two independent experiments by broth microdilution assay according to CLSI recommendations for A. baumannii and EUCAST recommendations for K. pneumoniae (Clinical and Laboratory Standards Institute, 2016 European Committee on Antimicrobial Susceptibility European Committee on Antimicrobial Susceptibility Testing [EUCAST], 2016 ). The initial inoculum of 5 x 105 CFU/mL for each strain was used in microtiter plates V (Deltlab, Spain) in presence of colistin, niclosamide, or colistin plus niclosamide, and incubated for 16–18 h at 37°C. Escherichia coli ATCC 25922 was used as control strain.
3
Niclosamide Attenuates Autoimmune Nephritis
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MRL/lpr mice were purchased from SLC Inc. (Japan). C57BL/6 mice were purchased from OrientBio (Korea). Niclosamide (Sigma-Aldrich, St Louis, MO, USA) was resuspended in 0.5% methyl cellulose (Sigma-Aldrich, St Louis, MO, USA) for in vivo studies or in 5% DMSO for in vitro use. Female 10-week-old MRL/lpr mice received daily administration of vehicle (n = 7) or Niclosamide (n = 7; 100 mg/kg) for 7 weeks by oral gavage. All mice were sacrificed at 16 weeks of age. Female 8-week-old C57BL/6 mice were treated via epicutaneous application of 50 μg of the TLR7 agonist resiquimod (R848; Sigma-Aldrich) dissolved in 10 μl of acetone, with or without 100 mg/kg of Niclosamide daily for 4 weeks, or acetone alone as a control, to the right ear three times a week until euthanasia. All procedure of animal research were provided in accordance with the Laboratory Animals Welfare Act, the Guide for the Care and Use of Laboratory Animals and the Guidelines and Policies for Rodent experiment provided by the IACUC(Institutional Animal Care and Use Committee) in school of medicine, The Catholic University of Korea. (Approval numbers: CUMS-2018–0341-02 and 2018-0236-02).
4
Evaluating Drug Effects with Niclosamide and Clofazimine
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The effect of drugs was tested in the presence of 1 μM Niclosamide (Sigma-Aldrich, Burlington, MA, United States) or 5 μM Clofazimine (Sigma-Aldrich, Burlington, MA, United States) for 10 min. Niclosamide (Sigma-Aldrich, Burlington, MA, United States, N0560000) and Clofazimine (Sigma-Aldrich, Burlington, MA, United States, Y0000313) stocks were prepared at 10 mM by resuspending in DMSO. Fresh dilutions were prepared for each experiment. In all the experiments, DMSO (vehicle) was used in controls at the same concentration as in the drug treated samples.
5
Niclosamide Preparation and Application
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Niclosamide (Sigma-Aldrich, St. Louis, MI, USA) working solutions were prepared prior to each experiment and used immediately after preparation to avoid precipitation. More specifically, Niclosamide was first weighed and diluted into a 1:1 solution of methanol:acetone (Sigma-Aldrich, St. Louis, MI, USA), followed by intense vortexing to yield a homogenous solution of 20 mM. The working solution was subsequently prepared through serial dilutions in dimethyl sulfoxide (DMSO; Sigma-Aldrich, St. Louis, MI, USA) to a concentration of 10 μM. Adequate quantities of the working solution were then diluted in a serum-free culture medium to yield the 30 nM and 100 nM desired concentrations for SELM stimulation. The final concentration percentage of DMSO in our cell cultures was 0.003% and 0.01% for the 30 nM and 100 nM concentrations of Niclosamide, respectively. The same solvent used for Niclosamide dilution was used as blank in SELM stimulations with no effect on SELM mRNA transcription, collagen production, or migration (Supplementary Table S1 ).
6
Niclosamide Effects on Mouse Retina
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Four-week-old C57BL/6 male mice (weighting 13–15 g) and 10 weeks old APCMin mice received once-daily oral administration of 100 µL of 40 mg/mL niclosamide (200 mg/kg of body weight; Sigma) by gavage as the niclosamide group, whereas the other mice were treated with PBS by gavage as the control group. After 4 weeks of treatment, the mice were euthanized 24 hours after the last drug administration, and retina tissues were collected.
7
Niclosamide Preparation and Application
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Niclosamide was purchased from Calbiochem (Cat# 481909-1GM). For cell treatment, Niclosamide powder was prepared in dimethyl sulfoxide (DMSO, Sigma-Aldrich, Cat# D8418) at a concentration of 10 mmol/L and further diluted in cell culture medium for indicated working concentrations for the treatment of cells. For the animal study, Niclosamide powder was dissolved in PBS with 10% Tween-80 (Sigma-Aldrich, Cat# P4780) and 5% ethanol at a concentration of 2 mg/mL for intraperitoneal injection.
8
Niclosamide Preparation and Application
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Niclosamide was purchased from Calbiochem (Cat# 481909-1GM). For cell treatment, Niclosamide powder was prepared in dimethyl sulfoxide (DMSO, Sigma-Aldrich, Cat# D8418) at a concentration of 10 mmol/L and further diluted in cell culture medium for indicated working concentrations for the treatment of cells. For the animal study, Niclosamide powder was dissolved in PBS with 10% Tween-80 (Sigma-Aldrich, Cat# P4780) and 5% ethanol at a concentration of 2 mg/mL for intraperitoneal injection.
9
Wnt/β-catenin Pathway Inhibitor Assay
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Wnt/β-catenin pathway inhibitors FH535, ICRT14, IWP-2, and niclosamide were purchased from Sigma-Aldrich, USAFH535, ICRT14, IWP-2, and niclosamide (10, 25, and 50μM each) were dissolved in the 0.5% dimethyl sulfoxide (DMSO). The data recorded from the migrations and proliferation with Wnt/β-catenin pathway inhibitors were compared against control group (DMSO 0.5%).
10
Bacterial Strain Cultivation and Quorum Sensing Signaling
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The bacterial strains and clinical isolates used in this study are listed in Supplementary Tables 1 , 2 , respectively. The strains were routinely grown at 37°C with aeration in Luria-Bertani broth (LB) supplemented, when required, with 3-(N-morpholino)propane sulfonic acid (MOPS; pH 7.0) at the final concentration of 50 mM. Synthetic 3OC12-HSL was prepared at the concentration of 10 mM in ethyl acetate acidified with 0.1% (v/v) acetic acid, while synthetic PQS stock solution was prepared at the concentration of 20 mM in methanol. Synthetic QS signal molecules were kindly provided by Proff. Paul Williams and Miguel Camara (Centre for Molecular Sciences, University of Nottingham, United Kingdom). Niclosamide and clofoctol were purchased from Sigma-Aldrich and dissolved in dimethyl sulfoxide (DMSO) at 10 and 80 mM final concentration, respectively.
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