Cyp2b6

Manufactured by BD

Sourced in United States

CYP2B6 is a laboratory equipment product that functions as a tool for the analysis and study of the CYP2B6 enzyme. CYP2B6 is a member of the cytochrome P450 enzyme family, which are responsible for the metabolism of various endogenous and exogenous compounds. The CYP2B6 product provides researchers with a means to investigate the role and activity of this specific enzyme.

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Lab products found in correlation

Cyp2c9, by BD (5 mentions) Cyp2c19, by BD (5 mentions) Cyp2d6, by BD (5 mentions) Cyp3a4, by BD (5 mentions) Cyp1a2, by BD (4 mentions) Cyp1b1, by BD (4 mentions) Cyp2c8, by BD (4 mentions) Cyp2e1, 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) Amplex red reagent, by Thermo Fisher Scientific (1 mentions) Menadione, by Merck Group (1 mentions) Superoxide dismutase, by Merck Group (1 mentions) 5 5 dithiobis 2 nitrobenzoic acid, by Merck Group (1 mentions) Glucose 6 phosphate (g6p), by Roche (1 mentions) 7 ethoxyresorufin, by Merck Group (1 mentions) Horseradish peroxidase, by Merck Group (1 mentions) Coumarin, by Merck Group (1 mentions)

5 protocols using cyp2b6

Cytochrome P450 Enzyme Inhibition Assay

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CYP inhibition was performed with the recombinant enzymes CYP1A2, CYP2B6, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 obtained from BD Biosciences. Compounds for CYP inhibition testing were prepared as 10 mM stock solutions in DMSO. Isoform-specific substrates were incubated at 37 °C individually with P450 enzymes (Table 5) and a range of test compound concentrations (1.1, 3.3 and 10 μM) in duplicate. Each isoform was tested separately with one reference compound, a known positive control inhibitor. During preincubation, the 96-well black plates were scanned with a fluorescence plate reader to eliminate false results originating from the autofluorescence of the test compounds. At the end of the incubation, product formation was monitored with fluorescence detection. A decrease in the formation of the metabolite compared to the “no inhibition” control samples was used to calculate the IC₅₀ value.

Kaczorowska K., Stankiewicz A., Bugno R., Paluchowska M.H., Burnat G., Brański P., Cieślik P., Wierońska J.M., Milik M., Nowak M., Przybyłowicz A., Kozioł A., Hogendorf A., Hogendorf A.S., Kalinowska-Tłuścik J., Duszyńska B., Pilc A, & Bojarski A.J. (2023). Design and Synthesis of New Quinazolin-4-one Derivatives with Negative mGlu7 Receptor Modulation Activity and Antipsychotic-Like Properties. International Journal of Molecular Sciences, 24(3), 1981.

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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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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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