Journal: Acta Pharmacologica Sinica

Article Title: Alflutinib (AST2818), primarily metabolized by CYP3A4, is a potent CYP3A4 inducer

doi: 10.1038/s41401-020-0389-3

Figure Lengend Snippet: Inhibition of CYP activities in HLMs

Article Snippet: Bupropion and hydroxy bupropion were supplied by Toronto Research Chemicals Inc. (Toronto, Canada).

Techniques: Inhibition, Concentration Assay, Activity Assay

Journal: The AAPS journal

Article Title: Metabolic Profiling of Human Long-Term Liver Models and Hepatic Clearance Predictions from In Vitro Data Using Nonlinear Mixed-Effects Modeling.

doi: 10.1208/s12248-016-0019-7

Figure Lengend Snippet: Fig. 2. Metabolic activities of in vitro liver models. The metabolite formation rates (black symbols) of 11 diverse phase I and phase II enzyme markers were determined in 2D cultures of HepG2 (at day 4), iPSC-derived hepatocyte-like cells (iCell®, at day 4), HepaRG™(at day 7), HepatoPac® (at day 8), HμREL™(at day 8), and primary pooled cryopreserved hepatocytes in suspension cultures (at day 0). The metabolites were quantified by LC-MS/MS and the formation rates were derived for 1′-hydroxymidazolam (CYP3A4), dextrorphan (CYP2D6), 4-hydroxydiclofenac (CYP2C9), hydroxybupropion (CYP2B6), hydroxytacrine (CYP1A2), benzydamine N-oxide (FMO), daunorubicinol (AKR), 8-oxo-O6-benzylguanine (AO), N-acetyl-sulfamethazine (NAT2), SN-38 glucuronide (UGT1A1), 7-hydroxycoumarin glucuronide (UGT), and sulfate (SULT). Gray symbols indicate the lower limit of quantification for the studied metabolites (for details, see Table I). The line represents the metabolic activity index calculated by averaging the determined metabolite formation rates of the 11 enzyme markers (see Table II). For the HepatoPac® system, cryopreserved primary hepatocytes from a single donor were used (Lot 3121A, BioreclamationIVT, Lot TLQ). Primary pooled cryopreserved hepatocytes were employed for the HμREL™model (Lot HU1007, BioreclamationIVT, Lot YMD (five donors)) and for the suspension culture studies (BioreclamationIVT, Lots ECO (ten donors), URK (five donors), and JJR (ten donors)). The different lots of primary hepatocytes showed similar metabolic activities for the CYP and UGT/ SULT enzyme markers in the BioreclamationIVT lot characterization sheets (listed in Supplemental 6)

Article Snippet: SN-38 (S589950), SN-38 glucuronide (S589980), hydroxybupropion (H830675), benzydamine N-oxide (B209960), hydroxytacrine (A629900), daunorubicinol (D194510), andN-acetyl-sulfamethazine (A187850) were purchased from Toronto Research Chemicals (Toronto, Canada).

Techniques: In Vitro, Derivative Assay, Suspension, Liquid Chromatography with Mass Spectroscopy, Activity Assay

Journal: bioRxiv

Article Title: Bupropion inhibits serotonin type 3AB heteromeric channels at a physiologically relevant concentration

doi: 10.1101/709881

Figure Lengend Snippet: (A) Sample traces of oocytes expressing 5-HT 3A R or 5-HT 3AB R in response to 5-HT (∼EC 30 ) alone and in combination with bupropion. 5-HT-evoked inward currents (grey, 5-HT 3A = 0.3 μM, 5-HT 3AB = 2 μM) were used as the control current. Following, the 5-HT concentration was kept constant and co-applied with increasing concentrations of bupropion (5-HT 3A : 10 - 1000 μM, 5-HT 3AB : 30 - 4000 μM). 5-HT 3AB R bottom panel, bar graph showing that a low concentration of 30 µM bupropion in the presence of 2 µM 5-HT elicits potentiation (122.6±8.0%, n = 6) in 5-HT 3AB R (B) Currents were normalized to the control currents and yielded the following IC 50 values, 5-HT 3A R: IC 50 = 87.1 µM (n H = 1.28±0.07, n = 5, means±S.E.M.), 5-HT 3AB R: IC 50 = 866 µM (n H = 2.07±0.20, n = 7, means ± S.E.M.) (C) Oocytes expressing 5-HT 3A R and 5-HT 3AB R did not elicit an inward current in response to bupropion alone. Statistical significance was determined with paired t-test (*p ≤ 0.05 **p ≤ 0.01 ***p ≤ 0.001).

Article Snippet: Prospect, IL); bupropion hydro-chloride and hydroxybupropion (Toronto Research Chemicals, Inc. North York, Canada); serotonin (5-HT; serotonin creatinine sulfate monohydrate) from Acros Organics (New Jersey, NJ).

Techniques: Expressing, Control, Concentration Assay

Journal: bioRxiv

Article Title: Bupropion inhibits serotonin type 3AB heteromeric channels at a physiologically relevant concentration

doi: 10.1101/709881

Figure Lengend Snippet: (A) Sample traces of oocytes expressing 5-HT 3A R (black, left panel) and 5-HT 3AB R (green, right panel). The first 5-HT evoked currents were used as the control currents (grey bars, ∼EC 30 , 5-HT 3A R: 0.5 µM, 5-HT 3AB R: 2 µM) and was co-applied with bupropion (magenta bars, ∼IC 50 , 5-HT 3A R: 100 µM, 5-HT 3AB R: 1 mM). Following the stable 5-HT response, bupropion (∼EC 50 ) was perfused for 5 min before another co-application of 5-HT and bupropion (B) Same experimental design as in (B), but with hydroxybupropion (blue bars, ∼IC 50 , 5-HT 3A R: 100 µM, 5-HT 3AB R: 500 µM). (C) Quantification of fractional inhibition of currents when the oocyte was pre-incubated in bupropion (magenta) or hydroxybupropion (blue) normalized to the control current (100%). Pre-incubation significantly reduced current amplitudes for 5-HT 3A R (Bup: 76.1±3.20%, n = 5; HydroB: 93.0±2.50%, n = 6) and 5-HT 3AB R (Bup: 35.5±2.29%, n = 6; HydroB: 46.1±2.47%, n = 4) as compared to co-application. Statistical significance was determined with paired t-test (*p ≤ 0.05 **p ≤ 0.01 ***p ≤ 0.001).

Article Snippet: Prospect, IL); bupropion hydro-chloride and hydroxybupropion (Toronto Research Chemicals, Inc. North York, Canada); serotonin (5-HT; serotonin creatinine sulfate monohydrate) from Acros Organics (New Jersey, NJ).

Techniques: Expressing, Control, Inhibition, Incubation

Journal: bioRxiv

Article Title: Bupropion inhibits serotonin type 3AB heteromeric channels at a physiologically relevant concentration

doi: 10.1101/709881

Figure Lengend Snippet: (A)-(B) Shows sample traces of bupropion application (magenta bar) and the recovery times for 5-HT 3A R (left panel, black) and 5-HT 3AB R (right panel, green). (A) In two-electrode voltage clamp experiments, oocytes expressing 5-HT 3A R and 5-HT 3AB R showed a stable response to repeated applications of 0.8 μM and 5 μM 5-HT at −60 mV, with a minimal wash time of 2 min and 90 s between each application, for 5-HT 3A R and 5-HT 3AB R respectively. (B) The first 5-HT evoked response represents the control current for the recovery experiment. Bupropion (∼IC 200 , A: 400 μM, B: 4000 μM) was applied alone for 60 s at - 60 mV, followed by an immediate application of 5-HT. The grey and magenta bars represent the time of application of 5-HT and bupropion, respectively. Moving down the panel, the wash times after bupropion application were 0 s, 30 s, and 60 s. 5-HT 3AB R current lacked desensitization and was slowly recovering for the 0 s wash after the application of bupropion and resulted in the application of 5 μM 5-HT for 3.5 min before withdrawing the agonist. (C) Quantitative representation of current amplitudes and results in (B) (n=3). 5-HT 3A R was maximally reduced to 82.4±1.78% and 5-HT 3AB R to 18.1±2.19% of the control current after 60s exposure to IC 200 bupropion with 0 s and 10 s wash, followed by a stepwise recovery. All currents could be recovered to ∼95% after ∼7.5 min wash. Statistical significance was determined with paired t-test (*p ≤ 0.05 **p ≤ 0.01 ***p ≤ 0.001).

Article Snippet: Prospect, IL); bupropion hydro-chloride and hydroxybupropion (Toronto Research Chemicals, Inc. North York, Canada); serotonin (5-HT; serotonin creatinine sulfate monohydrate) from Acros Organics (New Jersey, NJ).

Techniques: Expressing, Control

Journal: bioRxiv

Article Title: Bupropion inhibits serotonin type 3AB heteromeric channels at a physiologically relevant concentration

doi: 10.1101/709881

Figure Lengend Snippet: (A) Sample traces of 5-HT 3A R and 5-HT 3AB R expressing oocytes (5-HT 3A R: left, black; 5-HT 3AB R: right, green) in response to 5-HT (∼EC 50 ; top and bottom traces, 5-HT 3A R: 0.8 µM; 5-HT 3AB R: 5.0 µM) in the absence and presence of bupropion (magenta traces, ∼IC 50 ; 5-HT 3A R: 100 µM; 5-HT 3AB R: 1mM) at different voltages. (B) Quantification of fractional inhibition, currents were normalized to the control currents at each voltage (n=4). Data is shown as mean±S.E.M. Statistical significance was determined with paired t-test (*p ≤ 0.05 **p ≤ 0.01 ***p ≤ 0.001).

Article Snippet: Prospect, IL); bupropion hydro-chloride and hydroxybupropion (Toronto Research Chemicals, Inc. North York, Canada); serotonin (5-HT; serotonin creatinine sulfate monohydrate) from Acros Organics (New Jersey, NJ).

Techniques: Expressing, Inhibition, Control

Journal: bioRxiv

Article Title: Bupropion inhibits serotonin type 3AB heteromeric channels at a physiologically relevant concentration

doi: 10.1101/709881

Figure Lengend Snippet: (A) Sample traces of oocytes expressing 5-HT 3A R (black, left panel) and 5-HT 3AB R (green, right panel) in response to 0.5, 1.0, and 5.0 μM 5-HT (grey bars) followed by the same concentrations co-applied with 20 μM bupropion. Following the initial exposure to the three 5-HT concentrations (control current), the oocytes were exposed to 20 μM bupropion for at least 2 min before co-application with the agonist. (B) Quantitative representation of current amplitudes and results in (A) (A: n=4, AB: n=5). Data is shown as mean±S.E.M. Statistical significance was determined with paired t-test (*p ≤ 0.05 **p ≤ 0.01 ***p ≤ 0.001).

Article Snippet: Prospect, IL); bupropion hydro-chloride and hydroxybupropion (Toronto Research Chemicals, Inc. North York, Canada); serotonin (5-HT; serotonin creatinine sulfate monohydrate) from Acros Organics (New Jersey, NJ).

Techniques: Expressing, Control