To evaluate the effect of itraconazole or lapatinib on cell proliferation, log phase OE33, FLO-1, KYSE70 and KYSE510 cells were plated in 6-well plates at 1×105 cells/well and cultured in cell culture medium supplemented with 2.5 μM itraconazole (Sigma), 2.5 μM lapatinib (Cayman Chemical) or DMSO for up to 6 days, with replacement of medium and drug every 24 hours. Every 48 hours, cell proliferation was measured by using a hemocytometer, and each experiment was performed in triplicate. The cell viability assay was performed using alamarBlue according to the manufacturer’s protocol (Thermo Fisher Scientific). OE33 and KYSE510 cells were plated in Corning 96-well black plates with clear bottom at 1×103 cells/well and cultured in cell culture medium supplemented with 2.5 μM itraconazole or DMSO for 72 hours. Fluorescence signal was measured at the designated timepoints under the fluorimeter (excitation at 544 nm and emission at 590 nm) using a POLARstar Omega Microplate Reader (BMG Labtech). Fluorescence signal was normalized to the cells treated with DMSO for 24 hours.
Itraconazole
Manufactured by Merck Group
Sourced in United States, Germany, France, United Kingdom, Brazil, Hungary, India
Itraconazole is a broad-spectrum antifungal agent used in the treatment of various fungal infections. It functions by inhibiting the synthesis of ergosterol, a critical component of the fungal cell membrane, thereby disrupting the integrity and function of the fungal cell.
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Lab products found in correlation
Fluconazole, by Merck Group (42 mentions)
Voriconazole, by Merck Group (40 mentions)
Amphotericin b, by Merck Group (39 mentions)
Posaconazole, by Merck Group (35 mentions)
Ketoconazole, by Merck Group (21 mentions)
Caspofungin, by Merck Group (20 mentions)
Clotrimazole, by Merck Group (15 mentions)
Miconazole, by Merck Group (10 mentions)
Terbinafine, by Merck Group (8 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (8 mentions)
Anidulafungin, by Pfizer (7 mentions)
Isavuconazole, by Merck Group (7 mentions)
Micafungin, by Astellas Pharma (5 mentions)
Micafungin, by Merck Group (5 mentions)
Nystatin, by Merck Group (5 mentions)
Voriconazole, by Pfizer (4 mentions)
Fbs superior, by Merck Group (4 mentions)
Natamycin, by Merck Group (4 mentions)
Verapamil, by Merck Group (4 mentions)
Fluoxetine, by Merck Group (4 mentions)
138 protocols using itraconazole
1
Drug Effects on Cell Proliferation
2
Evaluating Anti-Proliferative Effects of Itraconazole and Lapatinib
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To evaluate the effect of itraconazole or lapatinib on cell proliferation, log-phase OE33, FLO-1, KYSE70, and KYSE510 cells were plated in 6-well plates at 1 × 105 cells/well and cultured in cell culture medium supplemented with 2.5 μmol/L itraconazole (Sigma), 2.5 μmol/L lapatinib (Cayman Chemical), or DMSO for up to 6 days, with replacement of medium and drug every 24 hours. Every 48 hours, cell proliferation was measured by using a hemocytometer, and each experiment was performed in triplicate. The cell viability assay was performed using AlamarBlue according to the manufacturer's protocol (Thermo Fisher Scientific). OE33 and KYSE510 cells were plated in Corning 96-well black plates with clear bottom at 1 × 103 cells/well and cultured in cell culture medium supplemented with 2.5 μmol/L itraconazole or DMSO for 72 hours. Fluorescence signal was measured at the designated timepoints under the fluorimeter (excitation at 544 nm and emission at 590 nm) using a POLARstar Omega Microplate Reader (BMG Labtech). Fluorescence signal was normalized to the cells treated with DMSO for 24 hours.
3
Orthotopic and Subcutaneous Tumor Models
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All animal experiments were approved by the Institutional Animal Care and Use Committee of the National Taiwan University College of Medicine (Taipei, Taiwan; no. 20200014). Five‐week‐old female mice were obtained from the National Laboratory Animal Center, Taipei, Taiwan. For orthotopic models, SCA9 (5 × 105) and MTC‐Q1 cells (1 × 106) were injected into the tongues of Swiss Webster and C57BL/6 mice, respectively. For the subcutaneous model, MTC‐Q1 cells (2 × 106) were injected subcutaneously into the flanks of C57BL/6 mice. Mice were sacrificed when the tumor volume exceeded 20 mm or the body weight was reduced by ≥ 20%. To evaluate the effect of itraconazole and anti‐PD‐1 antibody treatment on tumor growth in a subcutaneous model, itraconazole (600 μg/mouse, Sigma Aldrich, St. Louis, MO, USA) or the solvent control was administered intraperitoneally every day. The IgG2a isotype (clone 2A3) and anti‐PD‐1 (clone RMP1‐14) controls were obtained from Bio X Cell (Lebanon, NH, USA) and intraperitoneally injected twice a week (300 μg/mouse).
4
Itraconazole-Induced Apoptosis and Autophagy Modulation
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The human colon adenocarcinoma cell line, COLO 205, was purchased from the Food Industry Research and Development Institute, and the colorectal cell line, HCT 116, was kindly gifted by Dr Tzu-Ming Jao from Kaohsiung Veterans General Hospital (Kaohsiung, Taiwan). Both cell lines were maintained in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (Sigma-Aldrich; Merck KGaA), 2 mM L-glutamine (Gibco; Thermo Fisher Scientific, Inc.) and 1% penicillin and streptomycin (Invitrogen; Thermo Fisher Scientific, Inc.), at 37°C with 5% CO2.
To determine the role of autophagy on itraconazole induced apoptosis in COLO 205 cells, 5 mM 3-MA (3-methyladenine, Sigma-Aldrich; Merck KGaA) was co-cultured with 50 µM itraconazole (Sigma-Aldrich; Merck KGaA) at 37°C for 24 h.
To determine the role of autophagy on itraconazole induced apoptosis in COLO 205 cells, 5 mM 3-MA (3-methyladenine, Sigma-Aldrich; Merck KGaA) was co-cultured with 50 µM itraconazole (Sigma-Aldrich; Merck KGaA) at 37°C for 24 h.
5
Itraconazole-Loaded Polymer Dispersions
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Itraconazole (Sigma-Aldrich), polyvinyl alcohol (PVA) Mw=72000, (Merck) , Tween 80, Tween 20, Span 60 (Merck), Eumulgin CO40 (BASF), Cremophor RH 60 (BASF), Pluronic F127 (BASF), propylene glycol (PG) (Merck), benzyl benzoate (BB) (Merck), methanol (Merck), dichloromethane (DCM) (Merck), ethanol (Merck). Sabouraud 4% Dextrose Agar (SDA) (Merck), Sodium Dihydrogen Phosphate Dodecahydrate (Merck). Deionized distilled water was prepared using an in-house distillation assembly.
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Itraconazole (Sigma-Aldrich), polyvinyl alcohol (PVA) Mw=72000, (Merck
6
Antifungal Susceptibility Testing of Malassezia spp.
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Antifungal susceptibility testing was performed using a broth microdilution method as described by Leong et al. (10 (link)). Briefly, 200× drug stock dilutions were prepared at a 2× concentration in fresh OptiMAL medium. Amphotericin B, terbinafine, clotrimazole, miconazole, itraconazole, fluconazole, voriconazole, and ketoconazole were purchased from Sigma-Aldrich, Singapore. Stock and drug plate dilutions were prepared in accordance with CLSI and EUCAST guidelines. Yeast inocula were obtained from 4- to 7-day-old strains of Malassezia spp. A 50-μl yeast inoculum was added to 50 μl of 2× concentrated antifungals to achieve a final cell density of 5 × 103 to 5 × 104 CFU/ml. A 10-μl portion of 2× yeast inoculum that had been diluted 10 times was also plated onto a modified Leeming-Notman agar plate and incubated for 4 to 7 days at 35°C for postverification of the CFU inoculum (10 to 100 colonies per 10 μl). Each assay was performed in triplicate plates for a single culture at every individual time point or reading.
7
Fungicide Inhibitors Evaluation
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All fatty acids were obtained from Sigma-Aldrich (Poole, UK), Fluconazole, itraconazole and ketoconazole were obtained from Sigma-Aldrich (Poole, UK) whilst ketaminazole was supplied by professor T.R. Holman (University of California, Santa Cruz, CA). Lanosterol, eburicol and obtusifoliol were supplied by professor W. D. Nes (Texas Tech University, Lubbock, TX).
8
Antifungal Drug Susceptibility Testing
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All antifungal drugs including terbinafine (TRB; purity ≥ 98%, SIGMA), itraconazole (ITC; purity ≥ 99%, SIGMA), cyclopirox (CLO; purity ≥ 99%, European Pharmacopoeia Reference Standard), and fluconazole (FLU; purity ≥ 98%, SIGMA) were purchased in powder form from Sigma Chemical Co., St. Louis, MO and prepared as outlined in the clinical and laboratory standards institute (CLSI) broth microdilution method M38-A2. The working concentration ranges of drugs were 0.0009785∼0.5 μg/ml for TRB, 0.03125∼16 μg/ml for ITC and CLO and 0.125∼64 μg/ml for FLU.
9
Antifungal Susceptibility of C. africana Isolates
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All C. africana isolates obtained herein were tested for in vitro susceptibility to amphotericin B, flucytosine, fluconazole, itraconazole, voriconazole, and posaconazole (Sigma, St. Louis, MO, USA), caspofungin (Merck & Co., Inc.), and micafungin (Astellas Pharma) by using broth microdilution method as described in CLSI document M27-A3 and M27-S4. The MIC values were read following 24 h of incubation and determined visually as the lowest concentration in which prominent decrease in turbidity was observed excepting for amphotericin B which is defined as the lowest concentration in which the absence of turbidity is observed. The recently revised CLSI clinical breakpoints (CBPs) values for C. albicans were used as reference [28 (link)]. Quality control was performed as recommended in CLSI documents using C. parapsilosis ATCC22019 and C. krusei ATCC 6258.
10
Itraconazole and Endothelial Cell Lines Protocol
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Itraconazole was obtained from Sigma-Aldrich Inc. (St. Louis, MO), and Sporanox (Itraconazole: oral solution) was purchased from Janssen-Clag Ltd. (Batch NO. BKB4000). Endothelial cell lines (HUVEC and SVEC4-10) were obtained from Gibco (Invitrogen, Carlsbad, CA92008, USA) and the American Type Culture Collection (ATCC, Manassas, VA, USA). SKOV3ip1 and HeyA8 were a gift from Dr. Anil K. Sood, Department of Cancer Biology, University of Texas M.D. Anderson Cancer Center, TX, USA. All ovarian cancer cells (SKOV3ip1 or HeyA8) were maintained in RPMI 1640 supplemented with 10% fetal bovine serum (FBS). HUVECs were maintained in M200 media, and SVEC4-10 (mouse endothelial) cells were maintained in DMEM media supplemented with 10% fetal bovine serum (FBS). All cells were maintained in 5% CO2 at 37 °C.
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