Cyp2c8

Manufactured by BD

Sourced in United States

CYP2C8 is a laboratory equipment product used for the analysis and measurement of CYP2C8 enzyme activity. CYP2C8 is a member of the cytochrome P450 enzyme family, which plays a role in the metabolism of various drugs and other xenobiotics. The product is designed to facilitate the study and characterization of CYP2C8 enzyme function and interactions.

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Lab products found in correlation

Cyp2c9, by BD (5 mentions) Cyp2c19, by BD (5 mentions) Cyp2d6, by BD (5 mentions) Cyp2e1, by BD (5 mentions) Cyp1a2, by BD (4 mentions) Cyp2b6, by BD (4 mentions) Cyp3a4, by BD (4 mentions) Cyp1b1, by BD (4 mentions) Cyp3a5, by BD (4 mentions) Cyp2a6, by BD (3 mentions) Cyp1a1, by BD (2 mentions) Nadph, by Merck Group (2 mentions) Acetylthiocholine chloride, by Merck Group (1 mentions) Tetraisopropyl pyrophosphoramide, by Merck Group (1 mentions) Menadione, by Merck Group (1 mentions) Superoxide dismutase, by Merck Group (1 mentions) Glucose 6 phosphate (g6p), by Roche (1 mentions) 7 ethoxyresorufin, by Merck Group (1 mentions) Recombinant human acetylcholinesterase, by Merck Group (1 mentions) Amplex red reagent, by Thermo Fisher Scientific (1 mentions)

5 protocols using cyp2c8

Recombinant Human Cytochrome P450 Enzymes

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Baculovirus-insect
cell-expressed human CYP1A1, CYP1A2, CYP1B1, CYP2A6, CYP2B6, CYP2C8,
CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A4, CYP3A5, and CYP3A7 were purchased
from BD Biosciences Discovery Labware (Woburn, MA, USA) and used according
to the manufacturer’s instructions.

Juvonen R.O., Ahinko M., Jokinen E.M., Huuskonen J., Raunio H, & Pentikäinen O.T. (2021). Substrate Selectivity of Coumarin Derivatives by Human CYP1 Enzymes: In Vitro Enzyme Kinetics and In Silico Modeling. ACS Omega, 6(17), 11286-11296.

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Cytochrome P450 Enzyme Assay Protocol

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Recombinant human acetylcholinesterase, superoxide dismutase, horseradish peroxidase, NADPH, menadione, acetylthiocholine chloride, 5,5′-dithiobis(2-nitrobenzoic acid), 7-ethoxyresorufin, tetraisopropyl pyrophosphoramide, and coumarin were purchased from Sigma (St. Louis, MO). Parathion, paraoxon, and diethylthiophosphate were from Chem Service Inc. (West Chester, PA). Amplex Red reagent was from Molecular Probes (Eugene, OR). Recombinant human CYPs (CYP1A1, CYP1A2, CYP1B1, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A4, CYP3A5 and CYP 3A7), and pooled human liver microsomes were purchased from BD Gentest (Woburn, MA). Recombinant human CPR, Vivid P450 substrates, 7-ethoxy-methyloxy-3-cyanocoumarin (EOMCC) and 7-benzyloxy-methyloxy-3-cyanocoumarin (BOMCC), 7-methoxy-4-trifluoromethyl coumarin and dibenzylfluorescein were from Life Technologies (Grand Island, NY). Glucose-6-phosphate and Glucose-6-phosphate dehydrogenase were from Roche Diagnostic (Indianapolis, IN).

Jan Y.H., Richardson J.R., Baker A.B., Mishin V., Heck D.E., Laskin D.L, & Laskin J.D. (2015). Vitamin K3 (menadione) redox cycling inhibits cytochrome P450-mediated metabolism and inhibits parathion intoxication. Toxicology and applied pharmacology, 288(1), 114-120.

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In vitro Liver Microsome and Cytosol Preparation

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Fluorochloridone, NADPH, and glutathione (GSH) were purchased from Sigma-Aldrich (St. Louis, MO). Human liver microsomes (HLM) from a pool of 33 donors (Catalog. 452161) were purchased from BD Gentest Corp (Woburn, MA). Human liver cytosols (HLC) from a pool of 46 donors (Catalog. HMCYPL) were purchased from GIBCO (ThermoFisher Scientific, MA). The recombinant enzymes CYP1A1, CYP1A2, CYP1B1, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A4, and CYP3A5, produced in baculovirus were purchased from BD Gentest. Pooled mouse liver microsomes (MLM) and cytosols (MLC) were prepared from male C57BL/6 mouse liver homogenates by differential ultracentrifugation.^{15 (link)}

Shi J., Xie C., Liu H., Krausz K.W., Bewley C.A., Zhang S., Tang L., Zhou Z, & Gonzalez F.J. (2016). Metabolism and Bioactivation of Fluorochloridone, a Novel Selective Herbicide, in Vivo and in Vitro. Environmental science & technology, 50(17), 9652-9660.

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Characterization of CYP-Mediated Metabolism

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Monkey liver S9 fraction was obtained from BD Bioscience Pharmingen (San Diego, CA). Trizma pre-set crystals, reduced β-nicotinamide adenine dinucleotide phosphate (NADPH), alamethicin, buspirone, potassium phosphate buffer, Krebs-Henseleit buffer powder, trypan blue, and uridine 5’-diphosphoglucuronic acid (UDPGA) were obtained from Sigma (St. Louis, MO). LC-MS grade acetonitrile (ACN), water, and methanol (MeOH) were purchased from EM Science (Gibbstown, N.J.). Deuterated acetonitrile and deuterated methanol were obtained from Cambridge Isotopes Laboratories Inc. (Andover, MA). NMR tubes were acquired from Kimble/Chase Life Science (Vineland, NJ). Human recombinant CYP (Supersomes) CYP3A4, CYP2C9, CYP2C19, CYP1A2, CYP2D6, CYP2C8, and CYP2E1 were purchased from BD Biosciences (San Jose, CA). Cryopreserved pooled mouse, rat, dog, monkey, and human hepatocytes together with hepatocyte thawing medium were obtained from Bioreclamation IVT (Baltimore, MD). The test article (diamine purine containing parent compound) was synthesized in Celgene.

Apuy J.L., Xiang C., Franc S., Hegde S.G., Hubbard R., Zhao J, & Moghaddam M.F. (2016). Formation of A Novel Purine Metabolite through CYP3A4 Bioactivation and Glutathione Conjugation. Drug Metabolism Letters, 10(2), 144-150.

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In Vitro Metabolic Profiling

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Ocedurenone and KBP-5314 (internal standard) were prepared at KBP Biosciences,. Pooled human plasma was collected from healthy participants (Shanghai Center for Drug Metabolism and Pharmacokinetics Research, China). Human liver microsomes (HLM, Lot No.: 34689), human recombinant CYP1A2 (Lot No.: 30974), CYP1B1 (Lot No.: 68977), CYP2A6 (Lot No.: 33769), CYP2B6 (Lot No.: 49736), CYP2C8 (Lot No.: 03459), CYP2C9 (Lot No.: 00621), CYP2C19 (Lot No.: 69449), CYP2D6 (Lot No.: 75311), CYP2E1 (Lot No.: 73026), CYP3A4 (Lot No.: 16022), CYP3A5 (Lot No.: 44743), and CYP4A11 (Lot No.: 31489) were all purchased from BD Gentest™ (USA). Human primary hepatocytes were purchased from XenoTech (Lots H949 and HC2-8, Kansas, USA) and In Vitro Technologies (Lot OHO, MD, USA). Suppliers of materials used in the in vitro assays for relevant drug-metabolizing enzymes or transporters are listed in Table S1. Other reagents were commercially available and of special reagent grade, high-performance liquid chromatography grade, liquid chromatography–mass spectrometry grade, or equivalent. Further details of in vitro materials and methods can be requested from KBP Biosciences.

Wang P., Liu J., Tan X., Yang F., McCabe J, & Zhang J. (2023). Pharmacokinetics and Drug–Drug Interaction of Ocedurenone (KBP-5074) in vitro and in vivo. European Journal of Drug Metabolism and Pharmacokinetics, 48(4), 397-410.

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