Intracellular accumulation of Praziquantel in T lymphoblastoid cell lines, CEM (parental) and CEMVBL(P-gp-overexpressing).

Gabriel Kigen,Geoffrey Edwards

doi:10.1186/s40360-016-0079-4

Gabriel Kigen, Geoffrey Edwards

Open Access

https://doi.org/10.1186/s40360-016-0079-4

Copy DOI

Abstract

BackgroundPraziquantel (PZQ) is an antihelminthic drug whose P-glycoprotein (P-gp) substrate specificity has not been conclusively characterized. We investigated its specificity by comparing its in vitro intracellular accumulation in CEM (parental), and CEMvbl cells which over express P-gp, a drug efflux transporter. Saquinavir (SQV), a known substrate of efflux transporters was used as control.MethodsA reversed phase liquid chromatography method was developed to simultaneously quantify PZQ and SQV in cell culture media involving involved a liquid - liquid extraction followed by ultra-high performance liquid chromatography using a Hypurity C18 column and ultraviolet detection set at a wavelength of 215 nm. The mobile phase consisted of ammonium formate, acetonitrile and methanol (57:38:5 v/v). Separation was facilitated via isocratic elution at a flow rate of 1.5 ml/min, with clozapine (CLZ) as internal standard. This was validated over the concentration range of 1.6 to 25.6 μM for all analytes. Intracellular accumulation of SQV in CEMvbl was significantly lower compared to that in CEM cells (0.1 ± 0.031 versus 0.52 ± 0.046, p = 0.03 [p <0.05]).ResultsAccumulation of PZQ in both cell lines cells were similar (0.05 ± 0.005 versus 0.04 ± 0.009, p = 0.4) suggesting that it is not a substrate of P-gp in CEM cells. In presence tariquidar, a known inhibitor of P-gp, the intracellular accumulation of SQV in CEMvbl cells increased (0.52 ± 0.068 versus 0.61 ± 0.102, p = 0.34 in CEM cells and 0.09 ± 0.015 versus 0.56 ± 0.089, p = 0.029 [p < 0.05] in CEMvbl cells). PZQ did not significantly affect the accumulation of SQV in either CEM (0.52 ± 0.068 versus 0.54 ± 0.061, p = 0.77), or in CEMvbl cells (0.09 ± 0.015 versus 0.1 ± 0.031, p = 0.89) cells compared to tariquidar, implying that PZQ is not an inhibitor of P-gp in CEMvbl cells.ConclusionsPZQ is neither a substrate nor an inhibitor of the efflux drug transporter P-gp in T-lymphoblastoid cells, CEM and CEMvbl. We also report a simple, accurate and precise method for simultaneous quantification of PZQ and SQV.

Highlights

Praziquantel (PZQ) is an antihelminthic drug whose P-glycoprotein (P-gp) substrate specificity has not been conclusively characterized
Praziquantel (PZQ) is a broad spectrum antihelminthic drug used in the mass treatment of lymphatic filariasis and schistosomiasis which afflicts over 243 million people [1, 2]
The reverse-phase analysis was initially performed with various mixtures of ammonium formate, Accuracy and precision The mean inter-day precision was within the range for both drugs with average Coefficient of variation (CV) of between 3.28 and 5.89 % for PZQ, and 3.67 and 9.32 for SQV (Table 2)

Summary

Introduction

Praziquantel (PZQ) is an antihelminthic drug whose P-glycoprotein (P-gp) substrate specificity has not been conclusively characterized. Praziquantel (PZQ) is a broad spectrum antihelminthic drug used in the mass treatment of lymphatic filariasis and schistosomiasis which afflicts over 243 million people [1, 2]. It is widely used because of its low cost and efficacy. The coadministration may give rise to drug-drug interactions that could influence the treatment outcomes, which in some instances may require dosage adjustments in order to prevent toxicity or resistance [10,11,12]. There is currently limited information regarding PZQ’s mechanisms of action at molecular level, and a lot more research is required in its pharmacokinetics in order to prevent resistance [13]

Objectives

Methods

Results

Conclusion