Journal: Frontiers in Pharmacology

Article Title: Contributions of UDP-Glucuronosyltransferases to Human Hepatic and Intestinal Metabolism of Ticagrelor and Inhibition of UGTs and Cytochrome P450 Enzymes by Ticagrelor and its Glucuronidated Metabolite

doi: 10.3389/fphar.2021.761814

Figure Lengend Snippet: CYPs-selective metabolic pathways inhibition mediated by ticagrelor-O-Glu.

Article Snippet: Reference standard of ticagrelor (purity 95.0%), 7-Ethyl-10-hydroxy-camptothecin (SN-38) glucuronide (purity 97%), chenodeoxycholic acid 24-acyl-β-D-glucuronide (purity 96%), N-Acetyl serotonin β-D-glucuronide (purity 97%), mycophenolic acid β-D-glucuronide (purity 96%), 3′-azide-3′-deoxythymidine β-D-glucuronide sodium salt (purity 97%), bupropion (purity 98%), hydroxy bupropion (purity >98%), amodiaquine (purity >95%), N-Desethyl amodiaquine hydrochloride (purity >95%), 4′-hydroxylation diclofenac (purity >95%), S-mephenytoin, S - 4 - hydroxy mephenytoin (purity >98%), dextromethorphan (purity >95%), dextrorphan tartrate salt (purity >98%), and furafylline (purity >95%, internal standard) were purchased from Toronto Research Chemicals (Toronto, ON, Canada).

Techniques: Inhibition

Journal: Frontiers in Pharmacology

Article Title: Contributions of UDP-Glucuronosyltransferases to Human Hepatic and Intestinal Metabolism of Ticagrelor and Inhibition of UGTs and Cytochrome P450 Enzymes by Ticagrelor and its Glucuronidated Metabolite

doi: 10.3389/fphar.2021.761814

Figure Lengend Snippet: Effects of ticagrelor-O-Glu on the activities of human CYPs enzymes. bupropion, amodiaquine, diclofenac, [S]-mephenytoin, dextromethorphan, and testosterone were used as the probe substrates for CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6 and CYP3A4, respectively. Each bar represents the mean ± S.D. of triple incubations. *, p < 0.05; ***, p < 0.001, ****, p < 0.0001. Each bar represents the mean ± S.D. of triple incubations.

Article Snippet: Reference standard of ticagrelor (purity 95.0%), 7-Ethyl-10-hydroxy-camptothecin (SN-38) glucuronide (purity 97%), chenodeoxycholic acid 24-acyl-β-D-glucuronide (purity 96%), N-Acetyl serotonin β-D-glucuronide (purity 97%), mycophenolic acid β-D-glucuronide (purity 96%), 3′-azide-3′-deoxythymidine β-D-glucuronide sodium salt (purity 97%), bupropion (purity 98%), hydroxy bupropion (purity >98%), amodiaquine (purity >95%), N-Desethyl amodiaquine hydrochloride (purity >95%), 4′-hydroxylation diclofenac (purity >95%), S-mephenytoin, S - 4 - hydroxy mephenytoin (purity >98%), dextromethorphan (purity >95%), dextrorphan tartrate salt (purity >98%), and furafylline (purity >95%, internal standard) were purchased from Toronto Research Chemicals (Toronto, ON, Canada).

Techniques:

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