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Frontiers in Pharmacology 2020Vonoprazan fumarate is a potassium-competitive acid blocker that was developed as a novel acid-suppressing drug for multiple indications. As a potential alternative to...
BACKGROUND
Vonoprazan fumarate is a potassium-competitive acid blocker that was developed as a novel acid-suppressing drug for multiple indications. As a potential alternative to proton-pump inhibitors, the determination of the drug-drug interactions is vital for further applications. Probe drug cocktails are a type of rapid, economical, and efficient approach for evaluating cytochrome P450 enzyme activities. Since vonoprazan is metabolized partly by cytochrome P450, cocktails were used to study CYP-based drug-drug interactions.
METHODS
This study was conducted both and . In the study of rat liver microsomes, ultra-performance liquid chromatography coupled to tandem mass spectrometry was utilized to assess the reversible inhibition of cytochrome P450 by vonoprazan by determining the concentration of probe drugs (phenacetin, bupropion, tolbutamide, dextromethorphan, midazolam, chlorzoxazone). The differences in the levels of probe drugs between the rat groups with or without vonoprazan administration were also tested in the rats.
RESULTS
analysis revealed that the IC values of midazolam, tolbutamide, dextromethorphan, and bupropion in rat microsomes were 22.48, 18.34, 3.62, and 3.68 μM, respectively, while chlorzoxazone and phenacetin displayed no inhibition. analysis revealed that midazolam, bupropion, dextromethorphan, and tolbutamide showed significant ( < 0.05) differences in distinct pharmacokinetic parameters after vonoprazan administration, while those of chlorzoxazone and phenacetin were not significantly different.
CONCLUSION
The and results indicated that vonoprazan can inhibit CYP3A4, CYP2C9, CYP2D6, and CYP2B6, suggesting that the coadministration of vonoprazan with cytochrome P450 substrates should be performed cautiously in clinical settings.
PubMed: 32116727
DOI: 10.3389/fphar.2020.00053 -
Drug Design, Development and Therapy 2020Calycosin (CAL), a type of O-methylated isoflavone extracted from the herb (AM), is a bioactive chemical with antioxidative, antiphlogistic and antineoplastic...
BACKGROUND
Calycosin (CAL), a type of O-methylated isoflavone extracted from the herb (AM), is a bioactive chemical with antioxidative, antiphlogistic and antineoplastic activities commonly used in traditional alternative Chinese medicine. AM has been shown to confer health benefits as an adjuvant in the treatment of a variety of diseases.
AIM
The main objective of this study was to determine whether CAL influences the cytochrome P450 (CYP450) system involved in drug metabolism.
METHODS
Midazolam, tolbutamide, omeprazole, metoprolol and phenacetin were selected as probe drugs. Rats were randomly divided into three groups, specifically, 5% Carboxymethyl cellulose (CMC) for 8 days (Control), 5% CMC for 7 days + CAL for 1 day (single CAL) and CAL for 8 days (conc CAL), and metabolism of the five probe drugs evaluated using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).
RESULTS
No significant differences were observed for omeprazole and midazolam, compared to the control group. and t values of only one probe drug, phenacetin, in the conc CAL group were significantly different from those of the control group ( h: 0.50±0.00 vs 0.23±0.15; control vs conc CAL). of tolbutamide was decreased about two-fold in the conc CAL treatment group (conc vs control: 219.48 vs 429.56, <0.001).
CONCLUSION
Calycosin inhibits the catalytic activities of CYP1A2, CYP2D6 and CYP2C9. Accordingly, we recommend caution, particularly when combining CAL as a modality therapy with drugs metabolized by CYP1A2, CYP2D6 and CYP2C9, to reduce the potential risks of drug accumulation or ineffective treatment.
Topics: Animals; Cytochrome P-450 Enzyme Inhibitors; Cytochrome P-450 Enzyme System; Drugs, Chinese Herbal; Isoflavones; Medicine, Chinese Traditional; Metoprolol; Midazolam; Omeprazole; Phenacetin; Rats; Tolbutamide
PubMed: 32099327
DOI: 10.2147/DDDT.S236221 -
BMC Complementary Medicine and Therapies Feb 2022Echinacoside (ECH) possesses a wide range of biological activity. This present study analyzes the effect of ECH on cytochrome P450 isozymes (CYPs) activities of human...
BACKGROUND
Echinacoside (ECH) possesses a wide range of biological activity. This present study analyzes the effect of ECH on cytochrome P450 isozymes (CYPs) activities of human liver microsomes.
METHODS
The effect of ECH on CYPs enzyme activities were studied using the enzyme-selective substrates phenacetin (1A2), chlorzoxazone (2E1), S-mephenytoin (2C19), testosterone (3A4), coumarin (2A6), diclofenac (2C9), paclitaxel (2C8), and dextromethorphan (2D6). The IC50 values for CYP1A2, CYP2E1, CYP2C19, and CYP3A4 isoforms were examined to express the strength of inhibition. Further, the inhibition of CYPs was checked for time-dependent or not, and then fitted with competitive or non-competitive inhibition models. The corresponding parameters were also obtained.
RESULTS
ECH caused inhibitions on CYP1A2, CYP2E1, CYP2C19 and CYP3A4 enzyme activities in HLMs with IC50 of 21.23, 19.15, 8.70 and 55.42 μM, respectively. The obtained results showed that the inhibition of ECH on CYP3A4 was time-dependent with the KI/K value of 6.63/0.066 min·μM. Moreover, ECH inhibited the activity of CYP1A2 and CYP2E1 via non-competitive manners (K = 10.90 μM and K = 14.40 μM, respectively), while ECH attenuated the CYP2C19 activity via a competitive manner (K = 4.41 μM).
CONCLUSIONS
The results of this study indicate that ECH inhibits CYP1A2, CYP2E1, CYP2C19 and CYP3A4 activities in vitro. In vivo and clinical studies are warranted to verify the relevance of these interactions.
Topics: Cytochrome P-450 Enzyme Inhibitors; Cytochrome P-450 Enzyme System; Glycosides; Humans; Isoenzymes
PubMed: 35180866
DOI: 10.1186/s12906-022-03517-0 -
Frontiers in Pharmacology 2022Pharmacokinetic characterization plays a vital role in drug discovery and development. Although involving numerous laboratory animals with error-prone, labor-intensive,...
Pharmacokinetic characterization plays a vital role in drug discovery and development. Although involving numerous laboratory animals with error-prone, labor-intensive, and time-consuming procedures, pharmacokinetic profiling is still irreplaceable in preclinical studies. With physiologically based pharmacokinetic (PBPK) modeling, the profiles of drug absorption, distribution, metabolism, and excretion can be predicted. To evaluate the application of such an approach in preclinical investigations, the plasma pharmacokinetic profiles of seven commonly used probe substrates of microsomal enzymes, including phenacetin, tolbutamide, omeprazole, metoprolol, chlorzoxazone, nifedipine, and baicalein, were predicted in rats using bottom-up PBPK models built with data alone. The prediction's reliability was assessed by comparison with pharmacokinetic data reported in the literature. The overall predicted accuracy of PBPK models was good with most fold errors within 2, and the coefficient of determination (R) between the predicted concentration data and the observed ones was more than 0.8. Moreover, most of the observation dots were within the prediction span of the sensitivity analysis. We conclude that PBPK modeling with acceptable accuracy may be incorporated into preclinical studies to refine investigations, and PBPK modeling is a feasible strategy to practice the principles of 3Rs.
PubMed: 35645843
DOI: 10.3389/fphar.2022.895556 -
PloS One 2023GL-V9, a new synthetic flavonoid derived from wogonin, has shown beneficial biological functions. In this study, accurate and sensitive UPLC-MS/MS methods were developed...
GL-V9, a new synthetic flavonoid derived from wogonin, has shown beneficial biological functions. In this study, accurate and sensitive UPLC-MS/MS methods were developed and validated for the quantification of GL-V9 and its glucuronide metabolite (5-O-glucuronide GL-V9) in Beagle dog plasma. The chromatographic separation was performed on a C8 column (ACE Excel 5 C8 50×3.0 mm) using 0.1% formic acid and acetonitrile were used as mobile phase. Mass detection was performed on a triple quadrupole tandem mass spectrometer equipped with an electrospray ionization (ESI) interface operating in positive ion mode. Quantitative analysis was performed in multiple reaction monitoring (MRM) mode with the transitions of m/z 410.2→126.1 for GL-V9, m/z 586.3→410.0 for 5-O-glucuronide GL-V9 and m/z 180.0→110.3 for phenacetin (internal standard), respectively. The calibration curves for GL-V9 and 5-O-glucuronide GL-V9 showed excellent linearity over the concentration range of 0.5-500 ng/mL with correlation coefficient greater than 0.99. The intra- and inter-day accuracies were within 99.86% to 109.20% for GL-V9 and 92.55% to 106.20% for 5-O-glucuronide GL-V9, respectively. The mean recovery was 88.64% ± 2.70% for GL-V9, and 92.31% ± 6.28% for 5-O-glucuronide GL-V9, respectively. The validated method was successfully applied to the pharmacokinetic study in Beagle dogs after oral and intravenous administration. The oral bioavailability of GL-V9 was approximately 2.47%~4.35% in Beagle dogs and reached steady state on the fifth day after repeated dosing.
Topics: Dogs; Animals; Tandem Mass Spectrometry; Chromatography, Liquid; Chromatography, High Pressure Liquid; Glucuronides; Flavonoids; Reproducibility of Results
PubMed: 37285365
DOI: 10.1371/journal.pone.0286467 -
Pharmaceutics Mar 2020Mertansine, a tubulin inhibitor, is used as the cytotoxic component of antibody-drug conjugates (ADCs) for cancer therapy. The effects of mertansine on uridine...
Mertansine, a tubulin inhibitor, is used as the cytotoxic component of antibody-drug conjugates (ADCs) for cancer therapy. The effects of mertansine on uridine 5'-diphospho-glucuronosyltransferase (UGT) activities in human liver microsomes and its effects on the mRNA expression of cytochrome P450s (CYPs) and UGTs in human hepatocytes were evaluated to assess the potential for drug-drug interactions (DDIs). Mertansine potently inhibited UGT1A1-catalyzed SN-38 glucuronidation, UGT1A3-catalyzed chenodeoxycholic acid 24-acyl-β-glucuronidation, and UGT1A4-catalyzed trifluoperazine -β-d-glucuronidation, with values of 13.5 µM, 4.3 µM, and 21.2 µM, respectively, but no inhibition of UGT1A6, UGT1A9, and UGT2B7 enzyme activities was observed in human liver microsomes. A 48 h treatment of mertansine (1.25-2500 nM) in human hepatocytes resulted in the dose-dependent suppression of mRNA levels of CYP1A2, CYP2B6, CYP3A4, CYP2C8, CYP2C9, CYP2C19, UGT1A1, and UGT1A9, with IC values of 93.7 109.1, 36.8 18.3, 160.6 167.4, 32.1 14.9, 578.4 452.0, 539.5 233.4, 856.7 781.9, and 54.1 29.1 nM, respectively, and decreased the activities of CYP1A2-mediated phenacetin -deethylase, CYP2B6-mediated bupropion hydroxylase, and CYP3A4-mediated midazolam 1-hydroxylase. These in vitro DDI potentials of mertansine with CYP1A2, CYP2B6, CYP2C8/9/19, CYP3A4, UGT1A1, and UGT1A9 substrates suggest that it is necessary to carefully characterize the DDI potentials of ADC candidates with mertansine as a payload in the clinic.
PubMed: 32131538
DOI: 10.3390/pharmaceutics12030220 -
The Journal of Toxicological Sciences 2022According to ICH S3A Q&A focusing on microsampling, its application should be avoided in main study animals for test drugs that could exacerbate hematological parameters...
According to ICH S3A Q&A focusing on microsampling, its application should be avoided in main study animals for test drugs that could exacerbate hematological parameters with frequent blood sampling. However, no study has reported the effects of microsampling on toxicity parameters of drugs known to induce hematological toxicity. Therefore, we assessed the toxicological effects of serial microsampling on rats treated with phenacetin as a model drug. In a common 28-day study, 50 µL of microsampling was performed at 6-time points on days 1 to 2 and 7-time points on days 27 to 28 from the jugular vein of Sprague Dawley rats. The study was performed independently by two organizations. The toxicological influence of microsampling was evaluated on body weight, food consumption, hematology, blood clinical chemistry, urine parameters, organ weights, and tissue pathology. Phenacetin treatments induced significant changes of various hematological parameters (including hemoglobin and reticulocytes), some organ weights (including liver and spleen), and some hematology-related pathological parameters in the liver, spleen and bone marrow. Meanwhile, serial microsampling exhibited minimal influence on the assessed parameters, although 20 parameters showed statistical differences mostly at one organization. The current results support the notion that serial 50 μL microsampling from the jugular vein had minimal impacts on overall toxicological profiles even in rats treated with a drug inducing hematological toxicity, but the potential adverse effect on certain parameters could not be fully excluded. Accordingly, this microsampling technique has possibility to be employed even for non-clinical rat toxicity studies using drugs with potentially hematological toxicity.
Topics: Animals; Blood Specimen Collection; Body Weight; Jugular Veins; Phenacetin; Rats; Rats, Sprague-Dawley; Spleen
PubMed: 35527007
DOI: 10.2131/jts.47.193 -
Molecules (Basel, Switzerland) Jan 2023Benzisothiazolinone (BIT), a biocide widely used as a preservative in household cleaning and personal care products, is cytotoxic to lung cells and a known skin allergen...
Benzisothiazolinone (BIT), a biocide widely used as a preservative in household cleaning and personal care products, is cytotoxic to lung cells and a known skin allergen in humans, which highlights the importance of assessing its toxicity and pharmacokinetics. In this study, a simple, sensitive, and accurate LC−MS/MS method for the quantification of BIT in rat plasma, urine, or tissue homogenates (50 μL) using phenacetin as an internal standard was developed and validated. Samples were extracted with ethyl acetate and separated using a Kinetex phenyl−hexyl column (100 × 2.1 mm, 2.6 μm) with isocratic 0.1% formic acid in methanol and distilled water over a run time of 6 min. Positive electrospray ionization with multiple reaction monitoring transitions of m/z 152.2 > 134.1 for BIT and 180.2 > 110.1 for phenacetin was used for quantification. This assay achieved good linearity in the calibration ranges of 2−2000 ng/mL (plasma and urine) and 10−1000 ng/mL (tissue homogenates), with r ≥ 0.9929. All validation parameters met the acceptance criteria. BIT pharmacokinetics was evaluated via an intravenous and dermal application. This is the first study that evaluated BIT pharmacokinetics in rats, providing insights into the relationship between BIT exposure and toxicity and a basis for future risk assessment studies in humans.
Topics: Humans; Rats; Animals; Chromatography, Liquid; Tandem Mass Spectrometry; Disinfectants; Phenacetin; Reproducibility of Results
PubMed: 36677902
DOI: 10.3390/molecules28020845 -
RSC Advances Mar 2020PI-103 is a phosphatidylinositol 3-kinase inhibitor that includes multiple receptor affinity modifications, and it is also a therapeutic drug candidate primarily for...
An investigation of the metabolic activity, isozyme contribution, species differences and potential drug-drug interactions of PI-103, and the identification of efflux transporters for PI-103--glucuronide in HeLa1A9 cells.
PI-103 is a phosphatidylinositol 3-kinase inhibitor that includes multiple receptor affinity modifications, and it is also a therapeutic drug candidate primarily for human malignant tumors. However, its metabolic fate and potential drug-drug interactions involving human cytochrome P450 (CYP) and UDP-glucuronosyltransferases (UGT) enzymes remain unknown. In this study, our results demonstrated that the intrinsic clearance (CL) values of oxidated metabolite (M1) in human liver microsomes (HLM) and human intestine microsomes (HIM) were 3.10 and 0.08 μL min mg, respectively, while PI-103 underwent efficient glucuronidation with CL values of 15.59 and 211.04 μL min mg for mono-glucuronide (M2) by HLM and HIM, respectively. Additionally, reaction phenotyping results indicated that CYP1A1 (51.50 μL min mg), 1A2 (46.96 μL min mg), and UGT1A1 (18.80 μL min mg), 1A7 (8.52 μL min mg), 1A8 (8.38 μL min mg), 1A9 (34.62 μL min mg), 1A10 (107.01 μL min mg) were the most important contributors for the oxidation and glucuronidation of PI-103. Chemical inhibition assays also suggest that CYP1A2 and UGT1A1, 1A9 play a predominant role in the metabolism of PI-103 in HLM. Significant activity correlations were detected between phenacetin--deacetylation and M1 ( = 0.760, = 0.004) as well as β-estradiol-3--glucuronide and M2 ( = 0.589, = 0.044), and propofol--glucuronidation and M2 ( = 0.717, = 0.009). Furthermore, the metabolism of PI-103 revealed marked species differences, and dogs, rats, mice and mini-pigs were not the appropriate animal models. Gene silencing of breast cancer resistance protein (BCRP) or multidrug resistance-associated protein (MRPs) transporter results indicated that M2 was mainly excreted by BCRP, MRP1 and MRP4 transporters. Moreover, PI-103 displayed broad-spectrum inhibition towards human CYPs and UGTs isozymes with IC values ranging from 0.33 to 6.89 μM. Among them, PI-103 showed potent non-competitive inhibitory effects against CYP1A2, 2C19, 2E1 with IC and values of less than 1 μM. In addition, PI-103 exhibited moderate non-competitive inhibition against UGT1A7, 2B7, and moderate mixed-type inhibition towards CYP2B6, 2C9 and UGT1A3. Their IC and values were 1.16-6.89 and 0.56-5.64 μM, respectively. In contrast, PI-103 could activate the activity of UGT1A4 in a mechanistic two-site model with a value of 13.76 μM. Taken together, PI-103 was subjected to significant hepatic and intestinal metabolism. CYP1A1, 1A2 and UGT1A1, 1A7, 1A8, 1A9, 1A10 were the main contributing isozymes, whereas BCRP, MRP1 and MRP4 contributed most to the efflux excretion of M2. Meanwhile, PI-103 had a potent and broad-spectrum inhibitory effect against human CYPs and UGTs isozymes. These findings could improve understanding of the metabolic fates and efflux transport of PI-103. The inhibited human CYP and UGT activities could trigger harmful DDIs when PI-103 is co-administered with clinical drugs primarily cleared by these CYPs or UGTs isoforms. Additional studies are required to evaluate the clinical significance of the data presented herein.
PubMed: 35497201
DOI: 10.1039/c9ra09906a -
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi =... Aug 2022Primary human hepatocytes (PHH) are the gold standard of human liver model for drug screening. However, a problem of culturing PHH is the rapid decline of cytochrome...
Primary human hepatocytes (PHH) are the gold standard of human liver model for drug screening. However, a problem of culturing PHH is the rapid decline of cytochrome P450 (CYP450) activity, which plays an important role in drug metabolism. In this study, thermo-responsive culture dishes were used to explore the conditions for murine embryonic 3T3-J2 fibroblasts to form cell sheet. Based on the cell sheet engineering technology, a three-dimensional (3D) "sandwich" co-culture system of 3T3-J2 cell sheet/PHH/collagen gel was constructed. The tissue structure and protein expression of the model section were observed by hematoxylin eosin staining and immunofluorescence staining respectively. Phenacetin and bupropion were used as substrates to determine the activity of CYP450. The contents of albumin and urea in the system were determined by enzyme linked immunosorbent assay (ELISA). The results showed that the complete 3T3-J2 cell sheet could be obtained when the cell seeding density was 1.5×10 /dish (35 mm dish) and the incubation time at low temperature was 60 min. Through cell sheet stacking, a 3D liver model was developed. Compared with the two-dimensional (2D) model, in the 3D model, the cell-cell and cell-matrix connections were tighter, the activities of cytochrome P450 CYP1A2 and cytochrome P450 CYP2B6 were significantly increased, and the secretion levels of albumin and urea were increased. These indexes could be maintained stably for 21 d. Therefore, cell sheet stacking is helpful to improve the level of liver function of 3D liver model. This model is expected to be used to predict the metabolism of low-clearance drugs in preclinical, which is of great significance for drug evaluation and other studies.
Topics: Albumins; Animals; Cytochrome P-450 Enzyme System; Hepatocytes; Humans; Liver; Mice; Urea
PubMed: 36008342
DOI: 10.7507/1001-5515.202108056