Development, validation and utility of an in vitro technique for assessment of potential clinical drug–drug interactions involving P-glycoprotein

Keogh, John P.; Kunta, Jeevan

doi:10.1016/j.ejps.2005.11.011

Cited by 103 publications

(84 citation statements)

References 39 publications

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“…Acceptance criteria are defined for selected standard compounds and measurements (e.g., viability, resistance, integrity). These criteria are used to assess the permeability assay's functionality (29).…”

Section: Methods Suitabilitymentioning

confidence: 99%

“…There are three stages which compromise method suitability (Table I), including method development (optimization, standardization), demonstrating assay suitability (IVIVC), and permeability classification of new drugs (23). The assay is first optimized and standardized for the parameters that influence its experimental outcome to increase predictivity and throughput (26)(27)(28)(29). Additionally, the assay can be characterized for the presence of functional active transporters (e.g., amino acids, di/tripeptides, monocarboxylic acids, organic anions and cations) and drug efflux mechanisms.…”

Section: Methods Suitabilitymentioning

confidence: 99%

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Application of Method Suitability for Drug Permeability Classification

Volpe

2010

AAPS J

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Abstract. Experimental models of permeability in animals, excised tissues, cell monolayers, and artificial membranes are important during drug discovery and development as permeability is one of several factors affecting the intestinal absorption of oral drug products. The utility of these models is demonstrated by their ability to predict a drug's in vivo intestinal absorption. Within the various permeability models, there are differences in the performance of the assays, along with variability in animal species, tissue sources, and cell types, resulting in a variety of experimental permeability values for the same drug among laboratories. This has led to a need for assay standardization within laboratories to ensure applicability in the drug development process. Method suitability provides a generalized approach to standardize and validate a permeability model within a laboratory. First, assay methodology is optimized and validated for its various experimental parameters along with acceptance criteria for the assay. Second, the suitability of the model is demonstrated by a rank order relationship between experimental permeability values and human extent of absorption of known model compounds. Lastly, standard compounds are employed to classify a test drug's intestinal permeability and ensure assay reproducibility and quality. This review will provide examples of the different aspects method suitability for in situ (intestinal perfusions), ex vivo (everted intestinal sacs, diffusion chambers), and in vitro (cell monolayers, artificial membranes) experimental permeability models. Through assay standardization, reference standards, and acceptance criteria, method suitability assures the dependability of experimental data to predict a drug's intestinal permeability during discovery, development, and regulatory application.KEY WORDS: artificial membranes; drug permeability; ex vivo perfusion; in situ perfusion; in vitro cell monolayers; method suitability.

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Section: Methods Suitabilitymentioning

confidence: 99%

Section: Methods Suitabilitymentioning

confidence: 99%

Application of Method Suitability for Drug Permeability Classification

Volpe

2010

AAPS J

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“…MDCKII-MDR1 cell line has been extensively employed to conduct P-glycoprotein (P-gp/ MDR1) mediated drug efflux studies (Guo et al, 2002;Williams et al, 2003;Keogh and Kunta, 2006;Rodriguez-Proteau et al, 2006). P-gp, a MDR gene product, is an ATP-dependent drug efflux pump initially described in cancer chemotherapy.…”

Section: Introductionmentioning

confidence: 99%

Functional characterization of peptide transporters in MDCKII–MDR1 cell line as a model for oral absorption studies

Agarwal

Jain

Pal

et al. 2007

International Journal of Pharmaceutics

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MDCKII-MDR1 cell line has been extensively selected as a model to study P-gp-mediated drug efflux. Recently, investigators have employed this cell line for studying influx of peptide prodrug derivatives of parent compounds which are P-gp substrates. Therefore, the objective of this study is to functionally characterize the peptide mediated uptake and transport of [ 3 H] Glycylsarcosine ([ 3 H] Gly-Sar), a model peptide substrate across MDCKII-MDR1 cells. [ 3 H] Gly-Sar uptake from apical (AP) and basolateral (BL) membranes was found to be time dependent and saturable. MichaelisMenten (Km) constants of [ 3 H] Gly-Sar uptake across the AP and BL directions in MDCKII-MDR1 cell line were found to be 457 ± 37 μM and 464 ± 85 μM respectively. Vmax values in AP and BL directions for the peptide transporters in MDCKII-MDR1 cell line were calculated to be 0.035 ± 0.001 and 0.35 ± 0.034 pmol/min/mg protein respectively. Uptake of [ 3 H] Gly-Sar was significantly inhibited in the presence of aminocephalosporins and ACE-Inhibitors, known substrates for peptide transporters in both the AP and BL directions. Permeability of [ 3 H] Gly-Sar in the BL direction was maximal at pH 4 as compared to pH 5, 6 and 7.4 whereas such permeability in the AP direction was optimal at pH 7.4. Transepithelial transport of [ 3 H] Gly-Sar in the AP-BL direction was significantly lower than from BL-AP direction at all observed pHs. No statistical difference was observed in the transepithelial permeability of [ 3 H] Gly-Sar across both AP and BL directions over 4-10 days of growth period. The present study indicates that peptide transporters are effectively involved in the bidirectional transport of Gly-Sar across MDCKII-MDR1 cell line; the BL peptide transporter can transport Gly-Sar at a greater rate as compared to the AP peptide transporter. Results from these studies suggest the application of MDCKII-MDR1 cell line as a rapid effective tool to study peptide mediated influx of compounds that may be substrates for both P-gp and peptide transporters.

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“…Similarly, the inhibitory affinity of the drug substance for an apical efflux transporter is determined by measuring either the A-B or the B-A transport of a suitable probe substrate after pre-incubation with increasing concentrations of the drug substance in both compartments: [184,185] * * * *…”

Section: Transcellular Transport Studiesmentioning

confidence: 99%

Intestinal transporters for endogenic and pharmaceutical organic anions: the challenges of deriving in-vitro kinetic parameters for the prediction of clinically relevant drug–drug interactions

Grandvuinet

Vestergaard

Rapin³

et al. 2012

Journal of Pharmacy and Pharmacology

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Objectives This review provides an overview of intestinal human transporters for organic anions and stresses the need for standardization of the various in‐vitro methods presently employed in drug–drug interaction (DDI) investigations. Key findings Current knowledge on the intestinal expression of the apical sodium‐dependent bile acid transporter (ASBT), the breast cancer resistance protein (BCRP), the monocarboxylate transporters (MCT) 1, MCT3‐5, the multidrug resistance associated proteins (MRP) 1–6, the organic anion transporting polypetides (OATP) 2B1, 1A2, 3A1 and 4A1, and the organic solute transporter α/β (OSTα/β) has been covered along with an overview of their substrates and inhibitors. Furthermore, the many challenges in predicting clinically relevant DDIs from in‐vitro studies have been discussed with focus on intestinal transporters and the various methods for deducting in‐vitro parameters for transporters (Km/Ki/IC50, efflux ratio). The applicability of using a cut‐off value (estimated based on the intestinal drug concentration divided by the Ki or IC50) has also been considered. Summary A re‐evaluation of the current approaches for the prediction of DDIs is necessary when considering the involvement of other transporters than P‐glycoprotein. Moreover, the interplay between various processes that a drug is subject to in‐vivo such as translocation by several transporters and dissolution should be considered.

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Development, validation and utility of an in vitro technique for assessment of potential clinical drug–drug interactions involving P-glycoprotein

Cited by 103 publications

References 39 publications

Application of Method Suitability for Drug Permeability Classification

Application of Method Suitability for Drug Permeability Classification

Functional characterization of peptide transporters in MDCKII–MDR1 cell line as a model for oral absorption studies

Intestinal transporters for endogenic and pharmaceutical organic anions: the challenges of deriving in-vitro kinetic parameters for the prediction of clinically relevant drug–drug interactions

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