Journal: Acta Pharmaceutica Sinica. B

Article Title: Transfer learning enhanced graph neural network for aldehyde oxidase metabolism prediction and its experimental application

doi: 10.1016/j.apsb.2023.10.008

Figure Lengend Snippet: Identification of hAOX substrates using a liver cytosol assay (oxidative transformation).

Article Snippet: The human liver cytosol (mixed sex; a pool of 50 donors; catalog no. H0610.C, lot no. 1610027; 30 males and 20 females) was purchased from Sekisui XenoTech.

Techniques: Transformation Assay, Inhibition, Positive Control

Journal: Drug Metabolism and Disposition

Article Title: Stereospecific Metabolism of R - and S -Warfarin by Human Hepatic Cytosolic Reductases

doi: 10.1124/dmd.117.075929

Figure Lengend Snippet: Inhibitor phenotyping of steady-state reduction of R- and S-warfarin by HLC150. Reactions were carried out with two substrate concentrations, 50 µM (A and B) and 500 µM (C and D), 0.25 mg/ml protein, and 1 mM NADPH with inhibitors for specific cytosol reductases at 37°C and pH 7.4. (A and C) depict selective inhibition of R-warfarin to RS-warfarin alcohol. (B and D) show selective inhibition of S-warfarin to SS-warfarin alcohol. Reaction activities were normalized to the average control rate measured for reactions in the absence of inhibition. Rates were reported as percentages of the control rate. The reported activities are averaged from a minimum of five individual experiments ±S.E. Asterisks indicate statistical significance of inhibition compared with control as calculated using Student’s t test: *P < 0.05; **P < 0.005; ***P < 0.0005).

Article Snippet: High-performance liquid chromatography (HPLC)-grade methanol, acetic acid, dimethylsulfoxide (DMSO), and NADPH were purchased from Thermo Fisher (Pittsburgh, PA), and pooled human liver cytosol (HLC150) was from BD Biosciences (Franklin Lakes, NJ).

Techniques: Inhibition

Journal: Frontiers in Pharmacology

Article Title: Metabolism of F18, a Derivative of Calanolide A, in Human Liver Microsomes and Cytosol

doi: 10.3389/fphar.2017.00479

Figure Lengend Snippet: F18 metabolites detected in HLM, RLM, DLM, MLM, and HLcy after incubation with a substrate concentration of 50 μM for 1 h.

Article Snippet: Pooled mixed-gender human liver microsomes (HLMs), male beagle dog liver microsomes (DLMs), male monkey liver microsomes (MLMs), pooled mixed-gender human liver cytosol (HLcy), recombinant human cytochrome P450 enzymes (CYP1A1, CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP2J2, CYP3A4, CYP4F2, CYP4F3, and CYP4A11), and recombinant human flavin-containing monooxygenases (FMO1, FMO3, and FMO5) were purchased from BD Gentest (Woburn, MA, USA).

Techniques: Incubation, Concentration Assay

Journal: Frontiers in Pharmacology

Article Title: Metabolism of F18, a Derivative of Calanolide A, in Human Liver Microsomes and Cytosol

doi: 10.3389/fphar.2017.00479

Figure Lengend Snippet: Percentage of inhibition of the formation of M3-1 and M3-2 by chemical inhibitors in HLM and HLcy.

Article Snippet: Pooled mixed-gender human liver microsomes (HLMs), male beagle dog liver microsomes (DLMs), male monkey liver microsomes (MLMs), pooled mixed-gender human liver cytosol (HLcy), recombinant human cytochrome P450 enzymes (CYP1A1, CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP2J2, CYP3A4, CYP4F2, CYP4F3, and CYP4A11), and recombinant human flavin-containing monooxygenases (FMO1, FMO3, and FMO5) were purchased from BD Gentest (Woburn, MA, USA).

Techniques: Inhibition

Journal: Journal of Agricultural and Food Chemistry

Article Title: 4″-Sulfation Is the Major Metabolic Pathway of Epigallocatechin-3-gallate in Humans: Characterization of Metabolites, Enzymatic Analysis, and Pharmacokinetic Profiling

doi: 10.1021/acs.jafc.2c02150

Figure Lengend Snippet: Representative LC–MS/MS extracted ion chromatogram of EGCG-sulfates generated by human cytosol or standards. EGCG was incubated with HLC, small intestinal cytosol (HIC), and 3′-phosphoadenosine-5′-phosphosulfate (PAPS). The supernatants and authentic standards were analyzed by LC–MS/MS operated in full-scan mode. Analyses were carried out using the extracted ion chromatogram (XIC) of the m / z 537.03445 ion with a 0.002 Da window. RT means retention time.

Article Snippet: EGCG-4′′-sulfate, EGCG-3′′-sulfate, EGCG-3′′-glucuronide, and EGCG-4′′-glucuronide standards (purity >94%) were synthesized as previously reported., Pooled human liver and intestinal cytosols and microsomes from 15–50 donors of either sex were obtained from Xenotech (Kansas City, KS, USA).

Techniques: Liquid Chromatography with Mass Spectroscopy, Generated, Incubation

Journal: Journal of Agricultural and Food Chemistry

Article Title: 4″-Sulfation Is the Major Metabolic Pathway of Epigallocatechin-3-gallate in Humans: Characterization of Metabolites, Enzymatic Analysis, and Pharmacokinetic Profiling

doi: 10.1021/acs.jafc.2c02150

Figure Lengend Snippet: Concentration-dependent sulfation, methylation, and glucuronidation of EGCG by human liver or small intestinal fractions. Various concentrations of EGCG were incubated with (A) HLC and PAPS, (B) human small intestinal cytosol and PAPS, (C) HLC and S -adenosyl methionine (SAM), (D) human small intestinal cytosol and SAM, (E) human liver microsomes and uridine 5′-diphosphoglucuronic acid (UDPGA), and (F) human small intestinal microsomes and UDPGA. Data are presented as the mean ± SD of three independent experiments. The kinetic curve at lower or higher concentrations is shown in Figure S4 .

Article Snippet: EGCG-4′′-sulfate, EGCG-3′′-sulfate, EGCG-3′′-glucuronide, and EGCG-4′′-glucuronide standards (purity >94%) were synthesized as previously reported., Pooled human liver and intestinal cytosols and microsomes from 15–50 donors of either sex were obtained from Xenotech (Kansas City, KS, USA).

Techniques: Concentration Assay, Methylation, Incubation