MICs were determined as previously described (5 (link)). Briefly, cells were grown overnight in the indicated testing condition and then diluted to 5 × 106 CFU/mL with 2-fold serial dilutions of cefuroxime (Sigma-Aldrich) in the same medium and incubated overnight without aeration at 37°C. For ticlopidine synergy assays, a final concentration of 32 μg/mL ticlopidine (Sigma-Aldrich) was included in media with 2-fold serial dilutions of cefuroxime. This ticlopidine concentration was established as optimum for these MIC assays based on extensive pilot testing. The MIC was scored as the first well in which visual turbidity was decreased compared to the no-drug control well.
Ticlopidine
Manufactured by Merck Group
Sourced in United States, Germany
Ticlopidine is a pharmaceutical ingredient used in the production of various lab equipment and medical devices. It is a platelet aggregation inhibitor that helps prevent the formation of blood clots. The core function of Ticlopidine is to serve as a key component in the manufacturing of specialized lab equipment and medical products. Further details on its intended use or application are not available.
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Lab products found in correlation
Sulfaphenazole, by Merck Group (12 mentions)
Ketoconazole, by Merck Group (9 mentions)
Quinidine, by Merck Group (9 mentions)
Furafylline, by Merck Group (7 mentions)
Dextromethorphan, by Merck Group (5 mentions)
Diclofenac, by Merck Group (5 mentions)
Acetonitrile, by Thermo Fisher Scientific (5 mentions)
α naphthoflavone, by Merck Group (4 mentions)
Phenacetin, by Merck Group (4 mentions)
Acetaminophen, by Merck Group (4 mentions)
Testosterone, by Merck Group (4 mentions)
Omeprazole, by Merck Group (3 mentions)
Tranylcypromine, by Merck Group (3 mentions)
Tolbutamide, by Merck Group (3 mentions)
Coumarin, by Merck Group (3 mentions)
Mephenytoin, by Merck Group (3 mentions)
Chlorzoxazone, by Merck Group (3 mentions)
Midazolam, by Merck Group (3 mentions)
Amodiaquine, by Merck Group (3 mentions)
Quercetin, by Merck Group (3 mentions)
21 protocols using ticlopidine
1
Cefuroxime MIC Determination with Ticlopidine Synergy
2
Validation of KB Cell Line Authenticity
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The human oral carcinoma cell line KB were obtained from the American Type Culture Collection. The KB cells was originally thought to originate from an oral epidermal carcinoma, but has now been shown to be a HeLa derivative (15 (link),16 (link)) and vincristine-selected cell line KB/VCR was subscribed by professor Lihong Hu from Shanghai Institute of Materia Medica of Chinese Academy of Sciences. All cell lines have been under cell line authentication service and its result showed cell lines could be used for research. Cells were maintained in DMEM supplemented with 10% fetal bovine serum, 100 units/ml penicillin and 100 µg/ml streptomycin at 37°C and 5% CO2. PPD12 was synthesized by our laboratory as previously reported (11 (link)). ADM, midazolam, testosterone, phenacetin, bupropion, amodiaquine, diclofenac, s-mephenytoin, dextromethorphan, Ketoconazol, α-Naphthoflavone, Ticlopidine, Quercetin, Sulfaphenazole, Ticlopidine, Quinidine, 13C2,15N-acetaminophen and d9−1′-hydroxybufuralol were purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany). Acetonitrile and methanol [high-performance liquid chromatography (HPLC) grade] were obtained from Thermo Fisher Scientific, Inc. (Waltham, MA, USA). All other chemicals were of the highest quality available. A TUNEL staining assay kit was purchased from Wuhan Boster Biological Technology, Ltd. (Wuhan, China).
3
In Vitro Metabolism of Cannabinoids
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THC and CBD were obtained from Sigma-Aldrich (Merck, Germany). Alpha-naphthoflavone, ticlopidine, ketoconazole, sulfaphenazole, methoxsalen, trimethoprim, quindine, nicotinamide adenine dinucleotide phosphate (NADPH), ethyl acetate, GSH, Cys, and formic acid were purchased from Sigma-Aldrich (St. Louis, MO, USA). The MLM and HLM systems were provided by Bioreclamationivt Inc. (Hicksville, NY, USA). Recombinant human P450 enzymes were purchased from Xenotecch, LLC (Kansas City, MO, USA). All organic reagents were of the highest commercially available grade.
4
Comprehensive Pharmacological Compound Database
5
Human Liver Microsome Characterization
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BFT and BF were purchased from Shanghai Winherb Medical Technology Company (Shanghai, China). ABT, furafylline, sulfaphenazole, clomethiazole, omeprazole, 8-methoxypsoralen, ticlopidine, CYP3cide, glucose-6-phosphate dehydrogenase, D -glucose-6-phosphate, and NADP+ were obtained from Sigma (St. Louis, MO, United States). Montelukast, quinidine, ketoconazole was purchased from Jianglai Biotechnology Co., Ltd. (Shanghai, China). The pooled HLMs (from 50 donors, lot no. X008067) were obtained from BioreclamationIVT (Baltimore, MD, United States). A panel of baculovirus expressed human P450s (CYP1A1, 1A2, 2A6, 2A13, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, 3A4, 3A5, 4F2, and 4F3), co-expressing NADPH-CYP reductase and cytochrome b5 were obtained from BD Gentest Corp (Woburn, MA, United States). All chemicals and solvents were of analytical grade.
6
Comparative Pharmacokinetics of PZQ Stereoisomers
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R‐PZQ, S‐PZQ, cis‐4‐hydroxy‐PZQ, and trans‐4‐hydroxy‐PZQ were a generous gift from Merck Serono. 5,5‐Diethyl‐1,3‐diphenyl‐2‐iminobarbituric acid was obtained from AstraZeneca. PZQ, sulfaphenazole, KTZ, quinidine, ticlopidine, furafylline, diazepam (DPZ), acetonitrile (ACN), β‐Nicotinamide adenine dinucleotide 2′‐phosphate reduced tetrasodium salt (NADPH), formic acid, potassium phosphate monobasic, and potassium phosphate dibasic were obtained from Sigma Chemical Co. All other reagents were of the highest obtainable grade.
7
Quantitative Analysis of Metabolic Enzymes
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The pooled human liver microsomes (HLMs) and human recombinant enzyme used in the incubation studies were purchased from BD Gentest Co. (Woburn, MA, USA). Coumarin, 7-hydroxyCoumarin, tolbutamide, 4-hydroxytolbutamide, (S)-mephenytoin, 4′-hydroxymephenytoin, metoprolol, α-hydroxymetoprolol, midazolam maleate, 1-hydroxymidazolam, propranolol, quinidine hydrochloride, tranylcypromine, fluconazole, ticlopidine, quinidine, ketoconazole, furafylline, and diethyldithiocarbamate were purchased from Sigma Chemicals (St. Louis, MO, USA). Formic acid, ethylic acid, ammonium formate, MgCl2, NADP+, glucose 6-phosphate, glucose 6-phosphate dehydrogenase, and potassium phosphate (monobic and dibaic) were chromatographic-grade chemicals purchased from Sigma Chemicals (St. Louis, MO, USA). Acetonitrile and methanol were of analytical grade and purchased from Sigma Chemicals (St. Louis, MO, USA). PrimeScript® RT reagent Kit With gDNA Eraser and SYBR® Premix DimerEraser™ were purchased from Takara Biotechnology (Dalian, China). Glycyrrhetinic acid (GA) was purchased from National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China).
8
Cytochrome P450 Inhibitor Characterization
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Mefenamic acid (≥99%), tacrine (≥99%), carvedilol (≥98%), nifedipine (≥98%), ellipticine, α-naphthoflavone (≥97%), ticlopidine (≥99%), 1-naphtol (≥99%), 2-naphtol (≥98%), 4-methoxy-benzaldehyde (≥98%), 2-(p-tolyl)ethylamine (≥97%) were purchased from Sigma-Aldrich (Schnelldorf, Germany); phenacetin was obtained from Brocades-ACF (Maarssen, the Netherlands). 7-Methoxyresorufin was synthesized by the method of Burke and Mayer [28 (link)] and final purity was assessed to be higher than 95%.
9
Analytical Method for Metabolic Profiling
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LKMS was provided by Henan Pulike Biological Engineering Co., Ltd. (Luoyang, China). LC-MS-grade methanol (MeOH), acetonitrile (ACN), and formic acid (FA) were purchased from Fisher Scientific (Pittsburgh, PA, USA). Ultrapure water was purified using a Milli-Q system (Millipore Corporation, Billerica, MA, USA). The following compounds were purchased from Sigma-Aldrich (St. Louis, MO, USA): PH, CAN, BP, TBD, DEX, CLN, and TS. ACE, HCAN, HBP, HTBD, HDEX, HCLN, and HTS were purchased from Beijing Huizhi Taikang Biological Technology (Beijing, China). α-Naphthoflavone, Pilocarpine, Ticlopidine, Sulfaphenazole, Quinidine, sodium diethyldithiocarbamate, and Ketoconazole were purchased from Sigma (St. Louis, MO, USA). Phosphate buffer (PBS) tablets were purchased from Beijing Solarbio Technology (Beijing, China). β-nicotinamide adenine dinucleotide phosphate was purchased from Fisher Scientific (Pittsburgh, PA, USA).
PH, CAN, BP, TBD, DEX, CLN, and TS stock solutions were prepared in ACN at 10 mg/mL. Stock solutions of ACE, HCAN, HBP, HTBD, HDEX, HCLN, and HTS were prepared in MeOH at a concentration of 10 μg/mL. The internal standard of aflatoxin B1 was prepared at 200 ng/mL in ACN. The specific probe substrates, metabolites, and inhibitors are presented inTable 4 .
PH, CAN, BP, TBD, DEX, CLN, and TS stock solutions were prepared in ACN at 10 mg/mL. Stock solutions of ACE, HCAN, HBP, HTBD, HDEX, HCLN, and HTS were prepared in MeOH at a concentration of 10 μg/mL. The internal standard of aflatoxin B1 was prepared at 200 ng/mL in ACN. The specific probe substrates, metabolites, and inhibitors are presented in
10
Synthesis and Characterization of Peptides
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Glutathione (GSH), MgCl2, NADPH, potassium phosphate, amlodipine, caffeine, clozapine, ibuprofen, nimesulide and ticlopidine were purchased from Sigma–Aldrich chemicals (Helsinki, Finland). Acetonitrile was from J.T. Baker (St. Louis MO, USA) and formic acid from Merck KGaA (Darmstadt, Germany). d -Isomer peptides (gly–tyr–pro–cys–pro–his–pro, gly–tyr–pro–ala–pro–his–pro and gly–tyr–arg–pro–cys–pro–his–lys–pro) were synthesized by GL Biochem (Shanghai, China) and had a purity of >95%.
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