300,000 C2C12 cells (ATCC® CRL-1772™) were cultured on 50 cm2 Petri dishes in DMEM-F12 culture medium supplemented with 10% FBS and 1% antibiotics. Cells were incubated at 37 °C and 5% CO2, until the 65% confluence was reached (approx. 48h). Six groups of cultured cells were formed and they were cultured with 1) FBS 10% (proliferation conditions, Control), 2) Horse serum (HS, 2%, positive control), 3) 1 μM of EC, 4) 1 μM of PXR's agonist (PCN) (Merck KGaA, Darmstadt, Germany), 5) the combination of 10 μM of the PXR's antagonist (Ketoconazole) (Merck KGaA, Darmstadt, Germany) and 1μM of EC (30min) or 6) 1 μM PCN every 2 days until the 12th day was reached. Every 2 days, photographs were taken. At the end of the assay, total protein was obtained as described above. Protein content was measured by Bradford's method. Myogenin was detected and quantified as a differentiation marker in each group, with Western Blot assay (three independent assays in triplicate), with the same characteristics as those described above, using the anti-myogenin antibody (sc52903).
Ketoconazole
Manufactured by Merck Group
Sourced in United States, Germany, India, Sao Tome and Principe, United Kingdom, Macao, Israel
Ketoconazole is a laboratory product manufactured by Merck Group. It is an antifungal agent used for research and development purposes. The core function of Ketoconazole is to inhibit the synthesis of ergosterol, a key component of fungal cell membranes.
Automatically generated - may contain errors
Lab products found in correlation
Quinidine, by Merck Group (37 mentions)
Fluconazole, by Merck Group (32 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (28 mentions)
Sulfaphenazole, by Merck Group (27 mentions)
Amphotericin b, by Merck Group (25 mentions)
Itraconazole, by Merck Group (21 mentions)
Dextromethorphan, by Merck Group (14 mentions)
Miconazole, by Merck Group (14 mentions)
Acetonitrile, by Thermo Fisher Scientific (14 mentions)
Furafylline, by Merck Group (13 mentions)
Phenacetin, by Merck Group (13 mentions)
Voriconazole, by Merck Group (13 mentions)
Clotrimazole, by Merck Group (13 mentions)
Testosterone, by Merck Group (12 mentions)
Diclofenac, by Merck Group (12 mentions)
Glucose 6 phosphate dehydrogenase, by Merck Group (12 mentions)
Streptomycin, by Merck Group (11 mentions)
Posaconazole, by Merck Group (11 mentions)
Formic acid, by Merck Group (11 mentions)
Ampicillin, by Merck Group (10 mentions)
234 protocols using ketoconazole
1
Myogenesis Modulation by ECand PXR Agonists
2
Cytochrome P450 Inhibition Assay
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High-performance liquid chromatography (HPLC) grade methanol, acetic acid, triethylamine, and other analytical grade solvents were procured from Merck (Mumbai, India). 0.45 μm syringe filter and membrane filter were obtained from Phenomenex (Torrance, CA) and Millipore (Billerica, MA), respectively. Tinosporaside was purchased from Sigma chemicals (Steinheim, Germany). Vivid® CYP450 Screening kit and Vivid® substrates were purchased from Invitrogen Drug Discovery Solutions, USA. 96-well black microplate was obtained from NUNC (Roskilde, Denmark). Quinidine and sulfaphenazole were procured from Sigma (Steinheim, Germany). Ketoconazole and α-naphthoflavone were obtained from Merck (Mumbai, India).
3
Reagents and Materials for Cell Culture
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Dulbecco’s Modified Eagle Medium nutrient mixture (DMEM) (Fisher Scientific, Grand Island, NY, USA), penicillin/streptomycin solution (pen-strep) (Fisher Scientific, Grand Island, NY, USA), fetal bovine serum (FBS) (Fisher Scientific, Grand Island, NY, USA), and amphotericin B were purchased from Invitrogen (Camarillo, CA, USA). Heparin and ketoconazole were purchased from EMD Millipore Corp. USA (Burlington MA, USA). (Sigma-Aldrich, St. Louis, MO, USA). Climbazole was purchased from Tokyo Chemical Industry Co., LTD (Toshima, Kita-Ku, Tokyo, Japan). Unless otherwise indicated, all other drugs were purchased from commercial sources.
4
Hospital Wastewater Characterization and Analysis
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Diclofenac sodium salt (NaDCF, 98%+), fluconazole (98%+), hydrochlorothiazide (98%+), chlorhexidine (99%+), ketoconazole (98%+), triclosan (Irgasan, 97%+), atrazine (analytical standard), aluminum–nickel alloy (purum, 50% Al basis, 50% Ni basis) and an aqueous solution of NaBH4 (12 wt.% in 14 M NaOH) were delivered by Merck Co. (Prague, Czech Republic). Deuterated chloroform (CDCl3) was purchased from Merck Co. (Prague, Czech Republic). Additional chemicals and solvents in p.a. quality were obtained from a local supplier (Lach-Ner Co., Neratovice, Czech Republic).
A sample of hospital wastewater was stabilized with HNO3 for AOX (adsorbable organically bound halogens) determination [36 ]. The parameters of hospital wastewater stabilized with HNO3 are: pH = 2.07; [NH4+] = 32.4 mg L−1; [Cl−] = 820.7 mg L−1; [Ca+2 + Mg+2] = 4.5 mM; [NO3−] = 1213.6 mg L−1; CODCr = 349 mg O2 L−1; AOX = 1.44 mg Cl L−1. The experiments were carried out using deionized water, except where noted otherwise. All operations and manipulations were conducted in the air.
A sample of hospital wastewater was stabilized with HNO3 for AOX (adsorbable organically bound halogens) determination [36 ]. The parameters of hospital wastewater stabilized with HNO3 are: pH = 2.07; [NH4+] = 32.4 mg L−1; [Cl−] = 820.7 mg L−1; [Ca+2 + Mg+2] = 4.5 mM; [NO3−] = 1213.6 mg L−1; CODCr = 349 mg O2 L−1; AOX = 1.44 mg Cl L−1. The experiments were carried out using deionized water, except where noted otherwise. All operations and manipulations were conducted in the air.
5
Cytotoxicity Assay with DMSO and MTT
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Dimethyl sulfoxide (DMSO), 3-(4′,5′-dimethylthiazol-2′-yl)-2,5-diphenyltetrazolium bromide (MTT), cyclosporin A (CsA), ketoconazole, digoxin, ethyl paraben, acetonitrile (liquid chromatography [LC] grade), methanol and ethyl acetate (LC grade) were purchased from Merck (Merck Ltd., Taiwan). Sorafenib was purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum, 100 IU/mL penicillin, 100 mg/mL streptomycin, and 1% nonessential amino acids were purchased from Biological Industries (Cromwell, CT, USA). Milli-Q plus water (Millipore, Bedford, MA, USA) was used for all preparations.
6
Lipid Biosynthesis Inhibition Protocol
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Squalene (B50732), lanosterol (B50780), cholesterol (B20272), and stigmasterol (B20314) were purchased from Yuanye Biotechnology; cycloartenol (C923402) was purchased from Macklin; and mevinolin (PHR1285), fosmidomycin (F8307), terbinafine (T8826), and ketoconazole (PHR1385) were purchased from Merck.
7
Evaluation of Furin Inhibitor MI-1851
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The furin inhibitor MI-1851 was available from the previous studies [16] (link), its chemical structure is demonstrated in
Fig. 1 . Human serum albumin (HSA), racemic warfarin, racemic naproxen, CypExpress™ 3A4 Cytochrome P450 human kit, ketoconazole, testosterone and 6β-hydroxytestosterone were purchased from Merck (Darmstadt, Germany).Fig. 1 ![]()
Chemical structure of the furin inhibitor MI-1851.
8
Cytochrome P450 Enzyme Screening
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TG was purchased from Sigma-Aldrich. Vivid® CYP450 screening kit and Vivid® substrates (7-benzyloxymethyloxy-3 cyanocoumarin [Cat. No. P2861], ethoxymethyloxy- 3-cyanocoumarin [Cat. No. P3024]) were purchased from Invitrogen Drug Discovery Solutions, USA. CYP3A4 (Cat. No. P2858) and CYP2D6 (Cat. No. P2972) blue screening kit included baculosome (respective isozymes and NADPH-P450 reductase); regeneration system (glucose-6-phosphate, glucose-6-phosphate dehydrogenase) and NADP+ were used for the study. 96-well black-microplate was obtained from NUNC (Roskilde, Denmark). Ketoconazole and quinidine were obtained from Merck (Mumbai, India).
9
Pharmaceutical Compound Characterization Protocol
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Active pharmaceutical ingredients of studied medicines, i.e., atenolol (Daroupakhsh, Iran), benzoic acid (Merck, Iran), carbamazepine (Arastoo, Iran), carvedilol (Salehan Shimi, Iran), Ibuprofen (Daana, Iran), ketoconazole (Arastoo, Iran), lamotrigine (Arastoo, Iran), phenothiazine (Merck, Germany), phenytoin (Alhavi, Iran), piroxicam (Zahravi, Iran) salicylic acid (Merck, Germany), sulfamethoxazole (Merck, Germany), and tadalafil (Osveh, Iran), were provided from pharmaceutical and chemical companies (Purity: > 99%). Ethanol (96% w/w) was purchased from Jahan Alcohol (Iran), and choline chloride (> 99%), glycerol (> 99%), and urea (> 99%) from Merck (Germany). Lab-made double distilled water was used for the preparation of solutions.
10
Synthesis and Characterization of N-Sulfonylhydrazone Prototypes
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N-sulfonylhydrazone
prototypes, LASSBio-1771 (1 ), LASSBio-1772 (2 ), LASSBio-1773 (3 ), LASSBio-1774 (4 ),
and the internal standard (IP), biphenyl l-4-carboxylate methyl, were
synthesized at the Laboratório de Avaliação
e Síntese de Substâncias Bioativas (LASSBio,
UFRJ, Brazil) with ≥98% pure by HPLC (see theSupplemental Data ). HPLC grade solvents and reagents, i.e., acetonitrile, methanol, and formic acid (96%), were
purchased from Tedia-Brazil. Type I grade water produced by an ultrapure
water purification system (Master System MS2000, Gehaka, Brazil) was
used in experiments and HPLC analysis. Bis(p-nitrophenyl)
phosphate sodium salt, D-glucose-6-phosphate, glucose-6-phosphate
dehydrogenase, β-nicotinamide adenine dinucleotide phosphate
(NADP+), magnesium chloride hexahydrate, quinidine, sulfaphenazole,
furafylline, p-nitrophenol, ketoconazole, bicinchoninic
acid (BCA) protein assay kit, and salts used for buffer preparation
were purchased from Sigma-Aldrich (St. Louis, MO).
prototypes, LASSBio-1771 (
and the internal standard (IP), biphenyl l-4-carboxylate methyl, were
synthesized at the Laboratório de Avaliação
e Síntese de Substâncias Bioativas (LASSBio,
UFRJ, Brazil) with ≥98% pure by HPLC (see the
purchased from Tedia-Brazil. Type I grade water produced by an ultrapure
water purification system (Master System MS2000, Gehaka, Brazil) was
used in experiments and HPLC analysis. Bis(p-nitrophenyl)
phosphate sodium salt, D-glucose-6-phosphate, glucose-6-phosphate
dehydrogenase, β-nicotinamide adenine dinucleotide phosphate
(NADP+), magnesium chloride hexahydrate, quinidine, sulfaphenazole,
furafylline, p-nitrophenol, ketoconazole, bicinchoninic
acid (BCA) protein assay kit, and salts used for buffer preparation
were purchased from Sigma-Aldrich (St. Louis, MO).
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