Pooled hlm

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Sourced in United States

Pooled HLMs are human liver microsomes that have been pooled from multiple donors. They are a type of in vitro laboratory equipment used for drug metabolism and pharmacokinetic studies.

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Pooled human liver microsomes (HLMs) (4 citations)

11 protocols using pooled hlm

Lysionotin and Testosterone Characterization

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Lysionotin (≥98%) and testosterone (≥98%) were obtained from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). The chemical structure of lysionotin is shown in Figure 1. d-Glucose-6-phosphate, glucose-6-phosphate dehydrogenase, corticosterone (≥98%), NADP⁺, phenacetin (≥98%), acetaminophen (≥98%), 4-hydroxymephenytoin (≥98%), 7-hydroxy Coumarin (≥98%), 4′-hydroxy diclofenac (≥98%), sulfaphenazole (≥98%), quinidine (≥98%), tranylcypromine (≥98%), chlorzoxazone (≥98%), 6-hydroxychlorzoxazone (≥98%), paclitaxel (≥98%), 6β-hydroxytestosterone (≥98%), clomethiazole (≥98%), and furafylline (≥98%) were obtained from Sigma Chemical Co. Montelukast (≥98%) was obtained from Beijing Aleznova Pharmaceutical (Beijing, China). Coumarin (≥98%), diclofenac (≥98%), dextromethorphan (≥98%), and ketoconazole (≥98%) were purchased from ICN Biomedicals. Pooled HLMs were purchased from BD Biosciences Discovery Labware. All other reagents and solvents were of analytical reagent grade.

Li Y., Qin J., Wu H., Xu Y., Zhang L., Su K., Cui Y, & Wang H. (2020). In vitro inhibitory effect of lysionotin on the activity of cytochrome P450 enzymes. Pharmaceutical Biology, 58(1), 695-700.

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Inhibition of Human Liver Enzymes

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Anemarsaponin BII (≥98%) and testosterone (≥98%) were purchased from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). d-Glucose-6-phosphate, glucose-6-phosphate dehydrogenase, corticosterone (≥98%), NADP⁺, phenacetin (≥98%), acetaminophen (≥98%), 4-hydroxymephenytoin (≥98%), 7-hydroxy Coumarin (≥98%), 4′-hydroxy diclofenac (≥98%), sulfaphenazole (≥98%), quinidine (≥98%), tranylcypromine (≥98%), chlorzoxazone (≥98%), 6-hydroxychlorzoxazone (≥98%), paclitaxel (≥98%), 6β-hydroxytestosterone (≥98%), clomethiazole (≥98%) and furafylline (≥98%) were obtained from Sigma Chemical Co. Montelukast (≥98%) was obtained from Beijing Aleznova Pharmaceutical (Beijing, China). Coumarin (≥98%), diclofenac (≥98%), dextromethorphan (≥98%), and ketoconazole (≥98%) were purchased from ICN Biomedicals. Pooled HLMs were purchased from BD Biosciences Discovery Labware. All other reagents and solvents were of analytical reagent grade.

Wang M., Jiang W., Zhou J., Xue X, & Yin C. (2020). Anemarsaponin BII inhibits the activity of CYP3A4, 2D6, and 2E1 with human liver microsomes. Pharmaceutical Biology, 58(1), 1064-1069.

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Peucedanol Metabolism Assay Protocol

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Peucedanol (≥98%, Figure 1) and testosterone (≥98%) were purchased from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). d-Glucose-6-phosphate, glucose-6-phosphate dehydrogenase, corticosterone (≥98%), NADP⁺, phenacetin (≥98%), acetaminophen (≥98%), 4-hydroxymephenytoin (≥98%), 7-hydroxy Coumarin (≥98%), 4′-hydroxy diclofenac (≥98%), sulfaphenazole (≥98%), quinidine (≥98%), tranylcypromine (≥98%), chlorzoxazone (≥98%), 6-hydroxychlorzoxazone (≥98%), paclitaxel (≥98%), 6β-hydroxytestosterone (≥98%), clomethiazole (≥98%), and furafylline (≥98%) were obtained from Sigma Chemical Co. Montelukast (≥98%) was obtained from Beijing Aleznova Pharmaceutical (Beijing, China). Coumarin (≥98%), diclofenac (≥98%), dextromethorphan (≥98%), and ketoconazole (≥98%) were purchased from ICN Biomedicals. Pooled HLMs were purchased from BD Biosciences Discovery Labware. All other reagents and solvents were of analytical reagent grade.

Zhang C., Li Y., Yin C., Zheng J, & Liu G. (2021). In vitro study on the effect of peucedanol on the activity of cytochrome P450 enzymes. Pharmaceutical Biology, 59(1), 935-940.

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Characterization of Drug Metabolic Enzymes

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Bergenin (≥98%) and testosterone (≥98%) were obtained from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). d-Glucose-6-phosphate, glucose-6-phosphate dehydrogenase, corticosterone (≥98%), NADP⁺, phenacetin (≥98%), acetaminophen (≥98%), 4-hydroxymephenytoin (≥98%), 7-hydroxy Coumarin (≥98%), 4′-hydroxy diclofenac (≥98%), sulphaphenazole (≥98%), quinidine (≥98%), tranylcypromine (≥98%), chlorzoxazone (≥98%), 6-hydroxychlorzoxazone (≥98%), paclitaxel (≥98%), 6β-hydroxytestosterone (≥98%), clomethiazole (≥98%) and furafylline (≥98%) were obtained from Sigma Chemical Co. (St. Louis, MO). Montelukast (≥98%) was obtained from Beijing Aleznova Pharmaceutical (Beijing, China). Coumarin (≥98%), diclofenac (≥98%), dextromethorphan (≥98%) and ketoconazole (≥98%) were purchased from ICN Biomedicals (Aurora, OH). Pooled HLMs were purchased from BD Biosciences Discovery Labware (Bedford, MD). All other reagents and solvents were of analytical reagent grade.

Dong G., Zhou Y, & Song X. (2018). In vitro inhibitory effects of bergenin on human liver cytochrome P450 enzymes. Pharmaceutical Biology, 56(1), 620-625.

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Cytochrome P450 Enzyme Assay Reagents

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DHM (≥98%) and testosterone (≥98%) were obtained from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). The chemical structure of DHM is shown in Figure 1. d-Glucose-6-phosphate, glucose-6-phosphate dehydrogenase, corticosterone (≥98%), NADP⁺, phenacetin (≥98%), acetaminophen (≥98%), 4-hydroxymephenytoin (≥98%), 7-hydroxy Coumarin (≥98%), 4'-hydroxy diclofenac (≥98%), sulphaphenazole (≥98%), quinidine (≥98%), tranylcypromine (≥98%), chlorzoxazone (≥98%), 6-hydroxychlorzoxazone (≥98%), paclitaxel (≥98%), 6β-hydroxytestosterone (≥98%), clomethiazole (≥98%) and furafylline (≥98%) were obtained from Sigma Chemical Co. (Chicago, IL). Montelukast (≥98%) was obtained from Beijing Aleznova Pharmaceutical (Beijing, China). Dextrorphan was purchased from Toronto Research Chemical (North York, Canada). Coumarin (≥98%), diclofenac (≥98%), dextromethorphan (≥98%) and ketoconazole (≥98%) were purchased from ICN Biomedicals (Seven Hills, Australia). 6α-Hydroxypaclitaxel and Pooled HLMs were purchased from BD Biosciences (Woburn, MA). All other reagents and solvents were of analytical reagent grade.

Liu L., Sun S., Rui H, & Li X. (2017). In vitro inhibitory effects of dihydromyricetin on human liver cytochrome P450 enzymes. Pharmaceutical Biology, 55(1), 1868-1874.

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Obtusofolin Chemical Characterization

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Obtusofolin was obtained from Chengdu Must Bio-Technology Co. (Chengdu, China). The required substrates and reagents were purchased from Sigma Chemical Co. (Chicago, USA). Pooled HLMs were obtained from BD Bioscience (Woburn, USA). The purity of used chemicals was over 98% and the other reagents were of at least analytical reagent grade.

Liu N., Chen P., Du X., Sun J, & Han S. (2021). In vitro inhibitory effect of obtusofolin on the activity of CYP3A4, 2C9, and 2E1. BMC Complementary Medicine and Therapies, 21, 218.

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Cytochrome P450 Enzyme Inhibition Assay

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Ganoderic acid A (≥98%) and testosterone (≥98%) were obtained from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). The chemical structure of GAA is shown in Figure 1. d-Glucose-6-phosphate, glucose-6-phosphate dehydrogenase, corticosterone (≥98%), NADP⁺, phenacetin (≥98%), acetaminophen (≥98%), 4-hydroxymephenytoin (≥98%), 7-hydroxy Coumarin (≥98%), 4′-hydroxy diclofenac (≥98%), sulfaphenazole (≥98%), quinidine (≥98%), tranylcypromine (≥98%), chlorzoxazone (≥98%), 6-hydroxychlorzoxazone (≥98%), paclitaxel (≥98%), 6β-hydroxytestosterone (≥98%), clomethiazole (≥98%), and furafylline (≥98%) were obtained from Sigma Chemical Co. Montelukast (≥98%) was obtained from Beijing Aleznova Pharmaceutical (Beijing, China). Coumarin (≥98%), diclofenac (≥98%), dextromethorphan (≥98%), and ketoconazole (≥98%) were purchased from ICN Biomedicals. Pooled HLMs were purchased from BD Biosciences Discovery Labware. All other reagents and solvents were of analytical reagent grade.

Xu S., Zhang F., Chen D., Su K., Zhang L, & Jiang R. (2020). In vitro inhibitory effects of ganoderic acid A on human liver cytochrome P450 enzymes. Pharmaceutical Biology, 58(1), 308-313.

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Detailed Characterization of Drug Metabolic Pathways

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Friedelin (≥98%) and testosterone (≥98%) were obtained from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). d-Glucose-6-phosphate, glucose-6-phosphate dehydrogenase, corticosterone (≥98%), NADP⁺, phenacetin (≥98%), acetaminophen (≥98%), 4-hydroxymephenytoin (≥98%), 7-hydroxy Coumarin (≥98%), 4′-hydroxy diclofenac (≥98%), sulfaphenazole (≥98%), quinidine (≥98%), tranylcypromine (≥98%), chlorzoxazone (≥98%), 6-hydroxychlorzoxazone (≥98%), paclitaxel (≥98%), 6β-hydroxytestosterone (≥98%), clomethiazole (≥98%), and furafylline (≥98%) were obtained from Sigma Chemical Co. Montelukast (≥98%) was obtained from Beijing Aleznova Pharmaceutical (Beijing, China). Coumarin (≥98%), diclofenac (≥98%), dextromethorphan (≥98%), and ketoconazole (≥98%) were purchased from ICN Biomedicals. Pooled HLMs were purchased from BD Biosciences Discovery Labware. All other reagents and solvents were of analytical reagent grade.

Wei J., Zhang H, & Zhao Q. (2018). In vitro inhibitory effects of Friedelin on human liver cytochrome P450 enzymes. Pharmaceutical Biology, 56(1), 363-367.

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Cytochrome P450 Enzyme Characterization

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Succinic acid (≥98%) and testosterone (≥98%) were obtained from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). The chemical structure of succinic acid is shown in Figure 1. d-Glucose-6-phosphate, glucose-6-phosphate dehydrogenase, corticosterone (≥98%), NADP+, phenacetin (≥98%), acetaminophen (≥98%), 4-hydroxymephenytoin (≥98%), 7-hydroxy Coumarin (≥98%), 4′-hydroxy diclofenac (≥98%), sulfaphenazole (≥98%), quinidine (≥98%), tranylcypromine (≥98%), chlorzoxazone (≥98%), 6-hydroxychlorzoxazone (≥98%), paclitaxel (≥98%), 6β-hydroxytestosterone (≥98%), clomethiazole (≥98%), and furafylline (≥98%) were obtained from Sigma Chemical Co (MO, USA). Montelukast (≥98%) was obtained from Beijing Aleznova Pharmaceutical (Beijing, China). Coumarin (≥98%), diclofenac (≥98%), dextromethorphan (≥98%), and ketoconazole (≥98%) were purchased from ICN Biomedicals (Costa Mesa, California). Pooled HLMs were purchased from BD Biosciences Discovery Labware. All other reagents and solvents were of analytical reagent grade.

Wang H., Xia B., Lin M., Wang Y., Sun B, & Li Y. (2020). Succinic acid inhibits the activity of cytochrome P450 (CYP450) enzymes. Pharmaceutical Biology, 58(1), 1159-1164.

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In Vitro Metabolism of Anamorelin

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Anamorelin was provided by Toronto Research Chemical, Inc. (North York, ON, Canada). Pooled HLM, c-DNA-expressing CYP3A4, and three human flavin-containing monooxygenases (FMO) isoforms (FMO1, FMO3, and FMO5) were obtained from BD Gentest Corp. (Woburn, MA, USA). Glucose 6-phosphate (G6P), β-NADP⁺, glucose 6-phosphate dehydrogenase (G6Pd), and ammonium formate were obtained from Sigma Chemical Co. (St. Louis, MO, USA). All chemicals were analytical grade and were used as received. HPLC-grade acetonitrile (ACN) and distilled water (DW) were purchased from J.T. Baker (Phillipsburg, NJ, USA). Formic acid was purchased from Junsei Chemical Co. (Chou-ku, Japan).

Kwak Y.B., Seo J.I, & Yoo H.H. (2023). Exploring Metabolic Pathways of Anamorelin, a Selective Agonist of the Growth Hormone Secretagogue Receptor, via Molecular Networking. Pharmaceutics, 15(12), 2700.

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