scholarly journals Expression of Cytochrome P450 Enzymes Is Induced by Flaxseed Enterolignans

2021 ◽  
Vol 5 (Supplement_2) ◽  
pp. 312-312
Author(s):  
Danielle Defries ◽  
Shiva Shariati ◽  
Heather Blewett ◽  
Michel Aliani
Keyword(s):  
Cytochrome P450 ◽  
Fold Increase ◽  
Cytochrome P450 Enzymes ◽  
Funding Sources ◽  
Pressure Lowering ◽  
Cyp Metabolism ◽  
Hepatocyte Spheroids ◽  
Post Hoc ◽  
Circulating Levels ◽  
Cyp3a4 Expression

Abstract Objectives The major flaxseed lignan, secoisolariciresinol diglucoside (SDG), is metabolized to bioactive enterolignans in the intestine and liver. Once formed, enterolignans are subject to hepatic conjugation, similar to cytochrome P450 (CYP)-mediated conjugation of many drugs. CYPs also metabolize food-derived phytochemicals; therefore, adding flaxseed to the diet may promote drug-nutrient interactions affecting the potency of medications. Clinical trials have shown blood pressure-lowering effects of flaxseed in hypertensive patients; however, it is unknown if this stems from direct actions of flaxseed bioactives or indirectly from increased circulating levels of anti-hypertensive (AH) medications resulting from competition for CYP-mediated conjugation and inactivation. Our objective was to determine if enterolignans produced from flaxseed SDG affect expression or activity of CYP enzymes that metabolize metoprolol (MET), a commonly-prescribed AH medication. Methods 3D human hepatocyte spheroids were treated with 25 μM MET or 1, 10, or 100 μM enterodiol (END) for 48 hours, after which RNA was extracted for qPCR analysis of expression of CYP2C9 and CYP3A4, two CYP isozymes involved in MET metabolism. Spheroids were also treated with the same concentrations of MET and END for 1 and 48 hours to measure MET and END metabolites using HPLC/LC-QTOF-MS analysis. Differences were tested with 1-way ANOVA with Tukey's post-hoc test. Results MET, 1 μM END, and 10 μM END all induced an approximate 2-fold increase in CYP2C9 expression, while 100 μM END induced a 4-fold increase (P < 0.05). CYP3A4 expression was induced 5-fold by MET, while 1 μM and 10 μM END produced only a 2-fold increase in CYP3A4 expression (P < 0.05). 100 μM END led to an almost 14-fold increase in CYP3A4 expression (P < 0.05). Compared to 1 hour of co-treatment with 25 μM MET + END, O-demethylmetoprolol in media was significantly lower after 48 hours (P < 0.05), suggesting competition between END and MET for CYP metabolism. Conclusions Our initial experiments in 3D human hepatic spheroids are the first to show that metabolites of flaxseed lignans induce expression of CYP2C9 and CYP3A4. The fact that flaxseed enterolignans induce the same CYPs as MET supports the hypothesis of shared metabolism and potential metabolic competition between enterolignans and AH medications. Funding Sources NSERC Discovery Grant.

scholarly journals Assessment of inhibitory effects on major human cytochrome P450 enzymes by spasmolytics used in the treatment of overactive bladder syndrome

2017 ◽  
Vol 9 (7) ◽  
pp. 163-177
Author(s):  
Dominik Dahlinger ◽  
Sevinc Aslan ◽  
Markus Pietsch ◽  
Sebastian Frechen ◽  
Uwe Fuhr
Keyword(s):  
Cytochrome P450 ◽  
Liver Microsomes ◽  
Fold Increase ◽  
Pharmacokinetic Data ◽  
Cytochrome P450 Enzymes ◽  
Trospium Chloride ◽  
Screening Experiments ◽  
Human Cytochrome

Background: The objective of this study was to examine the inhibitory potential of darifenacin, fesoterodine, oxybutynin, propiverine, solifenacin, tolterodine and trospium chloride on the seven major human cytochrome P450 enzymes (CYP) by using a standardized and validated seven-in-one cytochrome P450 cocktail inhibition assay. Methods: An in vitro cocktail of seven highly selective probe substrates was incubated with human liver microsomes and varying concentrations of the seven test compounds. The major metabolites of the probe substrates were simultaneously analysed using a validated liquid chromatography tandem mass spectrometry (LC-MS/MS) method. Enzyme kinetics were estimated by determining IC50 and Ki values via nonlinear regression. Obtained Ki values were used for predictions of potential clinical impact of the inhibition using a static mechanistic prediction model. Results: In this study, 49 IC50 experiments were conducted. In six cases, IC50 values lower than the calculated threshold for drug–drug interactions (DDIs) in the gut wall were observed. In these cases, no increase in inhibition was determined after a 30 min preincubation. Considering a typical dosing regimen and applying the obtained Ki values of 0.72 µM (darifenacin, 15 mg daily) and 7.2 µM [propiverine, 30 mg daily, immediate release (IR)] for the inhibition of CYP2D6 yielded a predicted 1.9-fold and 1.4-fold increase in the area under the curve (AUC) of debrisoquine (CYP2D6 substrate), respectively. Due to the inhibition of the particular intestinal CYP3A4, the obtained Ki values of 14 µM of propiverine (30 mg daily, IR) resulted in a predicted doubling of the AUC for midazolam (CYP3A4 substrate). Conclusions: In vitro/ in vivo extrapolation based on pharmacokinetic data and the conducted screening experiments yielded similar effects of darifenacin on CYP2D6 and propiverine on CYP3A4 as obtained in separately conducted in vivo DDI studies. As a novel finding, propiverine was identified to potentially inhibit CYP2D6 at clinically occurring concentrations.

scholarly journals Substrate-induced modulation of protein-protein interactions within human mitochondrial cytochrome P450-dependent system

2020 ◽  
Author(s):  
E.O. Yablokov ◽  
T.A. Sushko ◽  
L.A. Kaluzhskiy ◽  
A.A Kavaleuski ◽  
Y.V. Mezentsev ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Steroid Hormones ◽  
Protein Interactions ◽  
Fold Increase ◽  
Cytochrome P450 Enzymes ◽  
Protein Protein Interactions ◽  
Adrenal Steroid ◽  
Hormone Biosynthesis ◽  
Multiple Levels ◽  
And Function

AbstractSteroidogenesis is strictly regulated at multiple levels, as produced steroid hormones are crucial to maintain physiological functions. Cytochrome P450 enzymes are key players in adrenal steroid hormone biosynthesis and function within short redox-chains in mitochondria and endoplasmic reticulum. However, mechanisms regulating supply of reducing equivalents in the mitochondrial CYP-dependent system are not fully understood. In the present work, we aimed to estimate how the specific steroids, substrates, intermediates and products of multistep reactions modulate protein-protein interactions between adrenodoxin (Adx) and mitochondrial CYP11s. Using the SPR technology we determined that steroid substrates affect affinity and stability of CYP11s – Adx complexes in an isoform-specific mode. In particular, cholesterol induces a 4-fold increase in the rate of CYP11A1 – Adx complex formation without significant effect on dissociation (koff decreased ~1.5-fold), overall increasing complex affinity. At the same time steroid substrates decrease the affinity of both CYP11B1 – Adx and CYP11B2 – Adx complexes, predominantly reducing their stability (4-7 fold). This finding reveals differentiation of protein-protein interactions within the mitochondrial pool of CYPs, which have the same electron donor. The regulation of electron supply by the substrates might affect the overall steroid hormones production. Our experimental data provide further insight into protein-protein interactions within CYP-dependent redox chains involved in steroidogenesis.

Pharmacogenomics of Cytochrome P450 Enzymes in Tumours

2004 ◽  
Vol 2 (3) ◽  
pp. 243-254 ◽  
Author(s):  
Diane Downie ◽  
Patrick Rooney ◽  
Morag McFadyen ◽  
Graeme Murray
Keyword(s):  
Cytochrome P450 ◽  
Cytochrome P450 Enzymes

Engineering Cytochrome P450 Enzymes

2008 ◽  
Vol 21 (1) ◽  
pp. 220-231 ◽  
Author(s):  
Elizabeth M. J. Gillam
Keyword(s):  
Cytochrome P450 ◽  
Cytochrome P450 Enzymes

scholarly journals Digit Variability in Carotenoid Scores Obtained with the Veggie Meter: A Pilot Study (P02-001-19)

2019 ◽  
Vol 3 (Supplement_1) ◽  
Author(s):  
Velarie Ansu ◽  
Stephanie Dickinson ◽  
Alyce Fly
Keyword(s):  
Indiana University ◽  
Left Hand ◽  
Funding Sources ◽  
Right Hand ◽  
Period Data ◽  
The Mean ◽  
The Right ◽  
Post Hoc ◽  
Main Effects ◽  
Mean Square Errors

Abstract Objectives To determine which digit and hand have the highest and lowest skin carotenoid scores, to compare inter-and-intra-hand variability of digits, and to determine if results are consistent with another subject. Methods Two subjects’ first(F1), second(F2), third(F3) and fifth(F5) digits on both hands were measured for skin carotenoids with a Veggie Meter, for 3 times on each of 18 days over a 37-day period. Data were subjected to ANOVA in a factorial treatment design to determine main effects for hand (2 levels), digits (4), and days (18) along with interactions. Differences between digits were determined by Tukey's post hoc test. Results There were significant hand x digit, hand x day, digit x day, and hand x digit x day interactions and significant simple main effects for hand, digit, and day (all P < 0.001). Mean square errors were 143.67 and 195.62 for subject A and B, respectively, which were smaller than mean squares for all main effects and interactions. The mean scores ± SD for F1, F2, F3, and F5 digits for the right vs left hands for subject A were F1:357.13 ± 45.97 vs 363.74 ± 46.94, F2:403.17 ± 44.77 vs. 353.20 ± 44.13, F3:406.76 ± 43.10 vs. 357.11 ± 45.13, and F5:374.95 ± 53.00 vs. 377.90 ± 47.38. For subject B, the mean scores ± SD for digits for the right vs left hands were F1:294.72 ± 61.63 vs 280.71 ± 52.48, F2:285.85 ± 66.92 vs 252.67 ± 67.56, F3:268.56 ± 57.03 vs 283.22 ± 45.87, and F5:288.18 ± 34.46 vs 307.54 ± 40.04. The digits on the right hand of both subjects had higher carotenoid scores than those on the left hands, even though subjects had different dominant hands. Subject A had higher skin carotenoid scores on the F3 and F2 digits for the right hand and F5 on the left hand. Subject B had higher skin carotenoid scores on F5 (right) and F1 (left) digits. Conclusions The variability due to hand, digit, and day were all greater than that of the 3 replicates within the digit-day for both volunteers. This indicates that data were not completely random across the readings when remeasuring the same finger. Different fingers displayed higher carotenoid scores for each volunteer. There is a need to conduct a larger study with more subjects and a range of skin tones to determine whether the reliability of measurements among digits of both hands is similar across the population. Funding Sources Indiana University.

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