Volume 9(6): 528-529 (2017) - 528 J Bioequiv Availab, an open access journal ISSN: 0975-0851 Open Access Kulsirirat et al., J Bioequiv Availab 2017, 9:6 DOI: 10.4172/jbb.1000357 Research Article OMICS International Journal of Bioequivalence & Bioavailability J o u r n a l o f B i o e q u i v a l e n c e & B i o a v a i l a b i l i t y ISSN: 0975-0851 Keywords: P-glycoprotein (P-gp); Non-ionic surfactant; High- roughput screening; Bioluminescence; Pgp-Glo™ assay; Efflux transporters Introduction e number of poorly soluble and lipophilic drugs with limited oral bioavailability among the new drug entities entering formulation development has increased over the last years. In many cases lower oral bioavailability entails higher inter-individual variability in bioavailability [1]. is can result in an inadequate control of plasma concentrations and pharmacological effects. Besides solubility and gastric stability, metabolic processes as well as transporter-mediated efflux play an essential role in influencing oral bioavailability of drugs [2]. P-glycoprotein (P-gp), also known as MDR1 and ABCB1, is a 170 kDa integral plasma membrane protein that functions as an ATP-dependent drug efflux pump and plays an essential role in multi-drug resistance and certain adverse drug-drug interactions [3]. Its efflux mechanism involves the protein binding to the ATP and requires energy derived by the hydrolysis of ATP to ADP in the presence of adenosine-triphosphatase enzyme (ATPase) [4]. P-gp belongs to the ATP-binding cassette (ABC) transporter subfamily that combines a wide range of proteins found in a number of different species. ABC transport proteins mediate the transport of a diversity of structurally different substances like amino acids, ions, peptides and also a variety of drugs through cell membranes [4]. A common method to improve the oral bioavailability of drugs is the use of solubilizing agents like complex forming hydroxypropyl β cyclodextrin, co-solvents or surfactants in drug formulations. However, the knowledge about interactions between nonionic surfactants and the efflux transporter P-gp is limited up to date. In this study, to investigated whether non- ionic surfactant stimulated or inhibited the ATPase activity of P-gp using Pgp-Glo TM assay systems. Methods Preparation of buffers and solutions e Pgp-Glo™ assay systems contain the necessary reagents for performing luminescent P-glycoprotein (P-gp) ATPase assays. e ATP detection buffer was thawed and equilibrated to room temperature. en the ATP detection buffer was shaken briefly to resuspend any precipitate The Potential of Non-Ionic Surfactant Against P-Glycoprotein Efflux Transporters for Drug Development System Kulsirirat T 1 , Rukthong P 2 , Dechwongya P 1 and Sathirakul K 1,3* 1 Faculty of Pharmacy, Mahidol University, Bangkok, Thailand 2 Faculty of Pharmacy, Srinakharinwirot University, Nakhon-Nayok, Thailand 3 Drug Discovery and Development Center, Thammasart University (Rangsit campus), Pathum Thani, Thailand Abstract Polysorbates (Tweens) are widely used in pharmaceutical formulations as excipients, such as stabilizer or solubilizer, for poorly soluble drugs. The efflux transporters ABCB1 (P-glycoprotein, P-gp) play an essential role in limiting the oral bioavailability of drugs. P-gp is expressed in the intestinal epithelium, thereby, it significantly impairs the systemic absorption of various active substances. The present study is to investigate a new property of polysorbates including its ability to stimulate or inhibit the efflux transporter P-glycoprotein (P-gp) using Pgp-Glo™ assay systems. The Pgp-Glo TM assay systems are luminescent P-glycoprotein (P-gp) ATPase assays. It detected the effect of substance on recombinant human P-gp in a cell membrane fraction. The investigation showed that the concentration of 0.1% Tween 80 is an inhibitor of P-gp ATPase activity, whereas, the concentrations of 0.5% and 1% of Tween 80 demonstrated as a stimulator. All of these concentrations of Tween 20 were observed to be an inhibitory effect on P-gp ATPase activity. that may form aſter freezing. en the entire content of the bottle of ATP detection buffer was transferred to the amber bottle containing the lyophilized ATP detection substrate and swirled or inverted several times to obtain a homogeneous solution of ATP detection reagent. Finally, 2.5 fold of concentration of test compounds in Pgp-Glo™ assay buffer were prepared. Preperation of P-gp membranes e Pgp-Glo™ assay system detects the effects of compounds on recombinant human P-gp in a cell membrane fraction. e P-gp membranes were prepared rapidly at 37 ° C and were immediately kept on ice until they became ready to use. Unused P-gp membranes were dispensed into single-use aliquots and stored at -70 ° C. en 5 mg/ml membranes were diluted to 1.25 mg/ml with Pgp-Glo™ assay buffer. Performing the assay e Pgp-Glo™ assay systems were used to analyze the effects of polysorbates (Tweens) on the activity of P-gp. Preperations of P-gp reaction mixtures and ATP reagent were done as described above. 20 μl of polysorbates were prepared in different concentrations (0.1, 0.5 and 1% v/v), Verapamil (positive control as P-gp substrate, 0.5 mM) and sodium orthovanadate (P-gp inhibitor, 0.25 mM) were added into a white opaque 96-well plate. P-gp was then added into the well containing the test substances and then this well were incubated at 37°C for 40 min. e reactions were stopped and luminescence was initiated by adding *Corresponding author: Sathirakul K, Drug Discovery and Development center, Thammasart University (Rangsit campus), Pathum Thani, Thailand, Tel: +66863208895; E-mail: pyksk2001@yahoo.com.sg Received May 17, 2017; Accepted September 27, 2017; Published October 05, 2017 Citation: Kulsirirat T, Rukthong P, Dechwongya P, Sathirakul K (2017) The Potential of Non-Ionic Surfactant Against P-Glycoprotein Efflux Transporters for Drug Development System. J Bioequiv Availab 9: 528-529. doi: 10.4172/jbb.1000357 Copyright: © 2017 Kulsirirat T, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.