Highly Miniaturized Formats for In Vitro Drug Metabolism Assays Using Vivid® Fluorescent Substrates and Recombinant Human Cytochrome P450 Enzymes

Trubetskoy, Olga V.; Gibson, Jasmin; Marks, Bryan D.

doi:10.1177/1087057104269731

Cited by 78 publications

(57 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although microsomal assays are commonly used to evaluate the activity of P450 enzymes, fluorescent methods for determining P450 activity allow for high-throughput screening and more reproducible results [21][22][23][24][25] . Bell et al found that nearly 75% of marketed drugs yielded acceptable correlations between fluorogenic and HLM+LC-MS/MS assays for CYPs 3A4, 2D6, and 2C9, which supports the use of fluorogenic assays for rapid early risk binning based on IC 50 values [26] .…”

Section: Discussionmentioning

confidence: 99%

Pungent ginger components modulates human cytochrome P450 enzymes in vitro

Chen

Yue

et al. 2013

Acta Pharmacol Sin

View full text Add to dashboard Cite

Aim: Ginger rhizome is used worldwide as a spicy flavor agent. This study was designed to explore the potential effects of pungent ginger components, 6-, 8-, and 10-gingerol, on human cytochrome P450 (CYP450) enzymes that are responsible for the metabolism of many prescription drugs. Methods: The activities of human CYP2C9, CYP2C19, CYP2D6, and CYP3A4 were analyzed using Vivid P450 assay kits. The mRNA expression of CYP3A4 in human hepatocellular carcinoma cell line HepG2 was measured using quantitative real-time PCR assay. Results: All three gingerols potently inhibited CYP2C9 activity, exerted moderate inhibition on CYP2C19 and CYP3A4, and weak inhibion on CYP2D6. 8-Gingerol was the most potent in inhibition of P450 enzymes with IC 50 values of 6.8, 12.5, 8.7, and 42.7 μmol/L for CYP2C9, CYP2C19, CYP3A4, and CYP2D6, respectively. By comparing the effects of gingerols on CYP3A4 with three different fluorescent substrate probes, it was demonstrated that the inhibition of gingerols on CYP3A4 had no substrate-dependence. In HepG2 cells, 8-gingerol and 10-gingerol inhibited, but 6-gingerol induced mRNA expression of CYP3A4. Conclusion: 6-, 8-, and 10-gingerol suppress human cytochrome P450 activity, while 8-and 10-gingerol inhibit CYP3A4 expression. The results may have an implication for the use of ginger or ginger products when combined with therapeutic drugs that are metabolized by cytochrome P450 enzymes.

show abstract

Section: Discussionmentioning

confidence: 99%

Pungent ginger components modulates human cytochrome P450 enzymes in vitro

Chen

Yue

et al. 2013

Acta Pharmacol Sin

View full text Add to dashboard Cite

show abstract

“…The assays were performed according to the manufacturer's recommendations (20). The incubation times were chosen according to the CYP and CYP substrate combination, as described earlier (21). For CYP2D6 and its substrate Vivid 2D6 cyan, the fluorescence was recorded every minute between 0 to 60 min after the start of the assay.…”

Section: Methodsmentioning

confidence: 99%

In Vitro and In Vivo Drug Interaction Study of Two Lead Combinations, Oxantel Pamoate plus Albendazole and Albendazole plus Mebendazole, for the Treatment of Soil-Transmitted Helminthiasis

Cowan

Vargas

Keiser

2016

Antimicrob Agents Chemother

View full text Add to dashboard Cite

bThe current treatments against Trichuris trichiura, albendazole and mebendazole, are only poorly efficacious. Therefore, combination chemotherapy was recommended for treating soil-transmitted helminthiasis. Albendazole-mebendazole and albendazole-oxantel pamoate have shown promising results in clinical trials. However, in vitro and in vivo drug interaction studies should be performed before their simultaneous treatment can be recommended. Inhibition of human recombinant cytochromes P450 (CYPs) CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 was tested by exposure to albendazole, albendazole sulfoxide, mebendazole, and oxantel pamoate, as well as albendazole-mebendazole, albendazole sulfoxide-mebendazole, albendazole-oxantel pamoate, and albendazole sulfoxide-oxantel pamoate. A high-pressure liquid chromatography (HPLC)-UV/visible spectroscopy method was developed and validated for simultaneous quantification of albendazole sulfoxide, albendazole sulfone, mebendazole, and oxantel pamoate in plasma. Albendazole, mebendazole, oxantel pamoate, albendazole-mebendazole, and albendazole-oxantel pamoate were orally applied to rats (100 mg/kg) and pharmacokinetic parameters calculated. CYP1A2 showed a 2.6-fold increased inhibition by albendazole-oxantel pamoate (50% inhibitory concentration [IC 50 ] ‫؍‬ 3.1 M) and a 3.9-fold increased inhibition by albendazole sulfoxide-mebendazole (IC 50 ‫؍‬ 3.8 M) compared to the single drugs. In rats, mebendazole's area under the concentration-time curve (AUC) and maximal plasma concentration (C max ) were augmented 3.5-and 2.8-fold, respectively (P ‫؍‬ 0.02 for both) when coadministered with albendazole compared to mebendazole alone. Albendazole sulfone was slightly affected by albendazole-mebendazole, displaying a 1.3-fold-elevated AUC compared to albendazole alone. Oxantel pamoate could not be quantified, translating to a bioavailability below 0.025% in rats. Elevated plasma levels of albendazole sulfoxide, albendazole sulfone, and mebendazole in coadministrations are probably not mediated by CYP-based drug-drug interaction. Even though this study indicates that it is safe to coadminister albendazole-oxantel pamoate and albendazole-mebendazole, human pharmacokinetic studies are recommended.A n estimated 465 million people are infected with the soiltransmitted helminth (STH) Trichuris trichiura, also known as human whipworm (1). Treatment programs (preventive chemotherapy) using the benzimidazoles albendazole and mebendazole are implemented to deworm patients infected with STHs. However, both drugs show low efficacy in single-dose treatment regimens (2, 3, 36).To make a step toward better treatment options for trichuriasis (4), combination therapies have received increased attention in the recent past. In more detail, in a recent study in Uganda, albendazole coadministered with mebendazole cured 54.2% of infections and reduced egg excretion by 94.3%, as opposed to low cure rates of single treatments with albendazole (15.4% cure rate; 54.9% egg reduction rate) and mebendazole (20.4% cur...

show abstract

“…These products can then be detected directly using a fluorescence plate reader. This method has the advantage that it is a fast and cost-effective way to perform thousands of IC 50 determinations per year (3,4). Measurements may be subject to interference from test inhibitors which either fluoresce or cause fluorescence quenching.…”

Section: In Vitro Methodologiesmentioning

confidence: 99%

In Vitro Evaluation of Reversible and Irreversible Cytochrome P450 Inhibition: Current Status on Methodologies and their Utility for Predicting Drug–Drug Interactions

Fowler

Zhang

2008

AAPS J

157

108

View full text Add to dashboard Cite

Abstract. It is widely accepted that today's practice of polypharmacy inevitably increases the incidence of drug-drug interactions (DDIs). Serious DDI is a major liability for any new chemical entity (NCE) entering the pharmaceutical market. As such, pharmaceutical companies employ various strategies to avoid problematic compounds for clinical development. A key cause for DDIs is the inhibition of cytochrome P450 enzymes (CYPs) that are responsible for metabolic clearance of many drugs. Screening for inhibition potency of CYPs by NCEs has therefore become a routine practice during the drug discovery stage. However, in order to make proper use of DDI data, an understanding of the strengths and weaknesses of the various experimental systems in current use is required. An illustrated review of experimental practices is presented with discussion of likely future developments. The combination of high quality in vitro data generation and the application of in vivo CYP inhibition modelling approaches should allow more informed decisions to be made in the search for drug molecules with acceptable DDI characteristics.

show abstract

Highly Miniaturized Formats for In Vitro Drug Metabolism Assays Using Vivid® Fluorescent Substrates and Recombinant Human Cytochrome P450 Enzymes

Cited by 78 publications

References 37 publications

Pungent ginger components modulates human cytochrome P450 enzymes in vitro

Pungent ginger components modulates human cytochrome P450 enzymes in vitro

In Vitro and In Vivo Drug Interaction Study of Two Lead Combinations, Oxantel Pamoate plus Albendazole and Albendazole plus Mebendazole, for the Treatment of Soil-Transmitted Helminthiasis

In Vitro Evaluation of Reversible and Irreversible Cytochrome P450 Inhibition: Current Status on Methodologies and their Utility for Predicting Drug–Drug Interactions

Contact Info

Product

Resources

About