Journal: Pharmacological Reports

Article Title: Interactions of fentanyl with blood platelets and plasma proteins: platelet sensitivity to prasugrel metabolite is not affected by fentanyl under in vitro conditions

doi: 10.1007/s43440-023-00447-7

Figure Lengend Snippet: Effect of fentanyl on platelet aggregation in whole blood. The influence of 2 or 2000 ng/ml fentanyl on whole blood platelet aggregation was measured in non-stimulated cells ( A ), cells stimulated with 2 µM ADP in combination with fentanyl ( B , C ), and cells stimulated with 5 µM ADP after 15-min preincubation with prasugrel metabolite and fentanyl ( D , E ). Data with normal distribution ( A , C ‒ E ) are expressed as mean ± SD, and those with non-normal distribution ( B ) are given as median with interquartile range, n = 6. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. control

Article Snippet: Prasugrel metabolite (R-138727) was obtained from BOC Sciences (Shirley, NY, USA).

Techniques: Control

Journal: Pharmacological Reports

Article Title: Interactions of fentanyl with blood platelets and plasma proteins: platelet sensitivity to prasugrel metabolite is not affected by fentanyl under in vitro conditions

doi: 10.1007/s43440-023-00447-7

Figure Lengend Snippet: The effect of fentanyl on ADP-induced platelet aggregation in PRP in the absence and presence of prasugrel metabolite

Article Snippet: Prasugrel metabolite (R-138727) was obtained from BOC Sciences (Shirley, NY, USA).

Techniques: Control, Incubation

Journal: Pharmacological Reports

Article Title: Interactions of fentanyl with blood platelets and plasma proteins: platelet sensitivity to prasugrel metabolite is not affected by fentanyl under in vitro conditions

doi: 10.1007/s43440-023-00447-7

Figure Lengend Snippet: Representative curves of ADP-induced platelet aggregation measured by LTA. The effect of fentanyl on agonist-induced platelet function was measured in PRP, in the absence or presence of 1.3 µM prasugrel metabolite (PM). The example data show platelet response to fentanyl added to PRP with subthreshold concentration of ADP ( A ) or preincubated with PM before stimulation of PRP with 5 µM ADP

Article Snippet: Prasugrel metabolite (R-138727) was obtained from BOC Sciences (Shirley, NY, USA).

Techniques: Concentration Assay

Journal: Pharmacological Reports

Article Title: Interactions of fentanyl with blood platelets and plasma proteins: platelet sensitivity to prasugrel metabolite is not affected by fentanyl under in vitro conditions

doi: 10.1007/s43440-023-00447-7

Figure Lengend Snippet: Effect of fentanyl on platelet activation. The influence of 2 or 2000 ng/ml fentanyl on platelet function was measured in blood, PRP or suspensions of isolated platelets in the following models: non-stimulated cells ( A ), cells stimulated with 2 µM ADP in combination with fentanyl ( B ), cells stimulated with 2 µM ADP after 15-min preincubation with fentanyl ( C ), and cells stimulated with 10 µM ADP after 15-min preincubation with prasugrel metabolite and fentanyl ( D ). Data with normal distribution ( A , B : PRP/platelets, C / D : blood/platelets, D : blood/PRP) are expressed as mean ± SD; otherwise data are presented as median with interquartile range, n = 6‒7. Before statistical analysis, data showing departure from normal distribution were log transformed for normalization. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. the appropriate controls: vehicle ( A ) or ADP ( B ‒ D )

Article Snippet: Prasugrel metabolite (R-138727) was obtained from BOC Sciences (Shirley, NY, USA).

Techniques: Activation Assay, Isolation, Transformation Assay

Journal: Pharmacological Reports

Article Title: Interactions of fentanyl with blood platelets and plasma proteins: platelet sensitivity to prasugrel metabolite is not affected by fentanyl under in vitro conditions

doi: 10.1007/s43440-023-00447-7

Figure Lengend Snippet: Summary of kinetic constants and binding affinities for the interactions of fentanyl and prasugrel metabolite with human plasma proteins assayed by SPR

Article Snippet: Prasugrel metabolite (R-138727) was obtained from BOC Sciences (Shirley, NY, USA).

Techniques: Binding Assay, Clinical Proteomics

Journal: Pharmacological Reports

Article Title: Interactions of fentanyl with blood platelets and plasma proteins: platelet sensitivity to prasugrel metabolite is not affected by fentanyl under in vitro conditions

doi: 10.1007/s43440-023-00447-7

Figure Lengend Snippet: Representative SPR sensograms showing the interactions of fentanyl or prasugrel metabolite with HSA, α 1 -AGP, ApoA-1, or ApoB-100. The proteins were immobilized to a dextran-coated gold surface. Injection of analyte (10, 20, 40 80 μM fentanyl or 2, 5, 10 μM prasugrel metabolite) produced a signal change that was directly proportional to the mass of bound drug molecules, expressed in resonance units (RU). Further details are given in

Article Snippet: Prasugrel metabolite (R-138727) was obtained from BOC Sciences (Shirley, NY, USA).

Techniques: Injection, Produced

Journal: Drug Metabolism and Disposition

Article Title: The Hydroxylation Position Rather than Chirality Determines How Efavirenz Metabolites Activate Cytochrome P450 46A1 In Vitro

doi: 10.1124/dmd.122.000874

Figure Lengend Snippet: Computational models of (S)-EFV (A) and L-Glu (B) binding to the allosteric sites on CYP46A1, showing some of the amino acid residues involved in the interaction with these compounds. The loop (from G417 to T426) separating the two allosteric sites is also shown and colored in magenta. The nitrogen, oxygen, chlorine, and fluorine atoms are in blue, red, light green, and cyan, respectively. The allosteric site mapping and compound docking were carried out in our previous work (Anderson et al., 2016; Mast et al., 2017a, 2020).

Article Snippet: All EFV metabolites were obtained from Toronto Research Chemicals (H805345, H941820, H941830, H941825, D452979, D46250 , D468240, D452800).

Techniques: Binding Assay

Journal: Drug Metabolism and Disposition

Article Title: The Hydroxylation Position Rather than Chirality Determines How Efavirenz Metabolites Activate Cytochrome P450 46A1 In Vitro

doi: 10.1124/dmd.122.000874

Figure Lengend Snippet: Phase 1 metabolic products of (S)-EFV generated by cytochrome P450 enzymes as suggested by previous studies (Avery et al., 2013b). The major P450s metabolizing (S)-EFV are shown in bold.

Article Snippet: All EFV metabolites were obtained from Toronto Research Chemicals (H805345, H941820, H941830, H941825, D452979, D46250 , D468240, D452800).

Techniques: Generated

Journal: Drug Metabolism and Disposition

Article Title: The Hydroxylation Position Rather than Chirality Determines How Efavirenz Metabolites Activate Cytochrome P450 46A1 In Vitro

doi: 10.1124/dmd.122.000874

Figure Lengend Snippet: Dependence of in vitro activation of CYP46A1 on the concentration of EFV (A), its mono- (B) and dihydroxylated (C) metabolites. The Y-axis is identical for all three graphs and represents CYP46A1 activity as nanomoles of 24-hydoxycholsterol (24HC) formed per nmole of CYP46A1 per minute. The results are the mean ±S.D. of the measurements from the three independent experiments. Statistically significant differences between (S)-EFV versus (R)-EFV were assessed by two-way ANOVA with Bonferroni multiple comparisons. No significant differences were found between the (S) enantiomer versus racemate of the same EFV metabolite * P ≤ 0.05.

Article Snippet: All EFV metabolites were obtained from Toronto Research Chemicals (H805345, H941820, H941830, H941825, D452979, D46250 , D468240, D452800).

Techniques: In Vitro, Activation Assay, Concentration Assay, Activity Assay

Journal: Drug Metabolism and Disposition

Article Title: The Hydroxylation Position Rather than Chirality Determines How Efavirenz Metabolites Activate Cytochrome P450 46A1 In Vitro

doi: 10.1124/dmd.122.000874

Figure Lengend Snippet: Effect of the allosteric site mutations on basal CYP46A1 activity and enzyme activation by L-Glu, EFV, and EFV metabolites. CYP46A1 activity is presented as nanomoles of 24-hydoxycholsterol (24HC) formed per nmole of CYP46A1 per minute. The results are the mean ±S.D. of the measurements from the three independent experiments. Statistically significant differences were assessed by one-way ANOVA with Tuckey multiple comparisons versus basal activity of WT or the CYP46A1 mutant. ** P ≤ 0.01; *** P ≤ 0.001.

Article Snippet: All EFV metabolites were obtained from Toronto Research Chemicals (H805345, H941820, H941830, H941825, D452979, D46250 , D468240, D452800).

Techniques: Activity Assay, Activation Assay, Mutagenesis

Journal: Drug Metabolism and Disposition

Article Title: The Hydroxylation Position Rather than Chirality Determines How Efavirenz Metabolites Activate Cytochrome P450 46A1 In Vitro

doi: 10.1124/dmd.122.000874

Figure Lengend Snippet: A summary of compound activation and binding to purified CYP46A1

Article Snippet: All EFV metabolites were obtained from Toronto Research Chemicals (H805345, H941820, H941830, H941825, D452979, D46250 , D468240, D452800).

Techniques: Activation Assay, Binding Assay, Purification

Journal: Drug Metabolism and Disposition

Article Title: The Hydroxylation Position Rather than Chirality Determines How Efavirenz Metabolites Activate Cytochrome P450 46A1 In Vitro

doi: 10.1124/dmd.122.000874

Figure Lengend Snippet: Effect of L-Glu or (S)-EFV on CYP46A1 activation by EFV metabolites. CYP46A1 activity is presented as nanomoles of 24-hydoxycholsterol (24HC) formed per nmole of CYP46A1 per minute. The results are the mean ±S.D. of the measurements from the three independent experiments. Statistically significant differences were assessed by one-way ANOVA with Tuckey multiple comparisons versus CYP46A1 activity when only compound in question was present. *** P ≤ 0.001.

Article Snippet: All EFV metabolites were obtained from Toronto Research Chemicals (H805345, H941820, H941830, H941825, D452979, D46250 , D468240, D452800).

Techniques: Activation Assay, Activity Assay

Journal: Drug Metabolism and Disposition

Article Title: The Hydroxylation Position Rather than Chirality Determines How Efavirenz Metabolites Activate Cytochrome P450 46A1 In Vitro

doi: 10.1124/dmd.122.000874

Figure Lengend Snippet: Spectral titrations of substrate-free CYP46A1 (A) and cholesterol-bound CYP46A1 (B) by EFV and some of its metabolites. Fit of spectral changes (ΔA, the amplitude of spectral response in the CYP46A1 difference spectrum) either to a hyperbolic equation or to the Hill equation, when cooperative binding was observed, is shown on the left, and the spectral response type in the CYP46A1 difference spectrum is shown on the right. The results are the mean ±S.D. of the measurements from the three independent titrations. Data for (S)-EFV are taken from (Mast et al., 2020).

Article Snippet: All EFV metabolites were obtained from Toronto Research Chemicals (H805345, H941820, H941830, H941825, D452979, D46250 , D468240, D452800).

Techniques: Binding Assay

Journal: The American Journal of Chinese Medicine

Article Title: Ultra-Performance Liquid Chromatography and Time-of-Flight Mass Spectrometry Analysis of Ginsenoside Metabolites in Human Plasma

doi: 10.1142/s0192415x11009470

Figure Lengend Snippet: Figure 2. UPLC/TOF-MS analysis of American ginseng saponins and metabolites. (A) Total ion chromatogram (TIC) of 15 standards of parent saponins and metabolites; (B) American ginseng root extract. Saponin peaks: 1, Rg1; 2, Re; 3, pseudoginsenoside F11; 4, Rh1; 5, Rg2; 6, 20(R)-Rg2; 7, Rb1; 8, Rc; 9, Rb2; 10, Rb3; 11, Rd; 12, Rg3; 13, 20(R)-Rg3; 14, C-K; and 15, Rh2; and (C) TOF mass chromatograms of ginsenoside Rb1, compound K (C-K) and internal standard (I.S.).

Article Snippet: Ginsenoside compound K (C-K) was obtained from ChromaDex.

Techniques:

Journal: The American Journal of Chinese Medicine

Article Title: Ultra-Performance Liquid Chromatography and Time-of-Flight Mass Spectrometry Analysis of Ginsenoside Metabolites in Human Plasma

doi: 10.1142/s0192415x11009470

Figure Lengend Snippet: Figure 3. Representative UPLC chromatograms of American ginseng saponins and metabolites in human plasma sample. (A) Total ion chromatogram (TIC) of a plasma sample. C-K was overlapped by other compounds. (B) Extracted ion chromatograms (XIC) of ginsenoside Rb1, C-K, and I.S. from the plasma matrix for quantification with a 0.01 Da mass window in negative modes. XIC at m/z 599.30–599.31 for Rb1, m/z 667.44–667.45 for C-K, and m/z 825.42–825.43 for I.S.

Article Snippet: Ginsenoside compound K (C-K) was obtained from ChromaDex.

Techniques: Clinical Proteomics