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P-glycoprotein Restricts Ocular Penetration of Loperamide across the Blood-Ocular Barriers: a Comparative Study in Mdr1a Knock-out and Wild Type Sprague Dawley Rats

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Abstract

The current research was undertaken to determine the existence and magnitude of P-glycoprotein (P-gp) expression on the blood-ocular barriers by studying the ocular penetration of loperamide, a specific P-gp substrate, in P-gp (Mdr1a) knock-out (KO) and wild type (WT) Sprague Dawley rats. A clear, stable, sterile solution of loperamide (1 mg/mL), for intravenous administration, was formulated and evaluated. Ocular distribution was studied in P-gp KO and WT rats following intravenous administration of loperamide (at two doses). The drug levels in plasma, aqueous humor (AH), and vitreous humor (VH) samples were determined with the aid of UHPLC-Q-TOF-MS/MS, and the AH/plasma (D AH ) and VH/plasma (D VH ) distribution ratios were estimated. Electroretinography (ERG), ultrastructural analyses, and histology studies were carried out, in both KO and WT rats, to detect any drug-induced functional and/or structural alterations in the retina. Dose-related loperamide levels were observed in the plasma of both WT and KO rats. The loperamide concentrations in the AH and VH of KO rats were significantly higher compared to that observed in the WT rats, at the lower dose. However, a marked increase in the D AH and D VH was noted in the KO rats. ERG, ultrastructure, and histology studies did not indicate any drug-induced toxic effects in the retina under the test conditions. The results from these studies demonstrate that P-gp blocks the penetration of loperamide into the ocular tissues from the systemic circulation and that the effect is more pronounced at lower plasma loperamide concentrations.

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Abbreviations

ABC, ATP-binding cassette; AH, Aqueous humor; BAB, Blood-aqueous barrier; BM, Bruch’s membrane; BOB, Blood-ocular barrier; BQ, Below quantification; BRB, Blood-retinal barrier; Ch, Choriocapillaris; D AH , Distribution ratio of loperamide from plasma to aqueous humor; D VH , Distribution ratio of loperamide from plasma to vitreous humor; EE, Extraction efficiency; ERG, Electroretinography; I.P., Intraperitoneal; IIG, Inactive ingredients; INL, Inner nuclear layer; IS, Inner segment; IV, intravenous; KO, Knock-out; LOD, Limit of detection; LOQ, Limit of quantification; Mdr1a, Multidrug resistance 1a gene; ONL, Outer nuclear layer; OS, Outer segment; P-gp, P-glycoprotein; PG, pigmented granules; Q-TOF, Quadrupole time of flight; RPE, Retinal pigmented epithelium; SD, Sprague Dawley; UHPLC, Ultra-high performance liquid chromatography; VH, Vitreous humor; WT, Wild type

Funding

This project was supported by the grant from the National Eye Institute, National Institutes of Health [Grant#-1R01EY022120-03]. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

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Author notes

  1. Akshaya Tatke and Karthik Yadav Janga contributed equally to this work.

Authors and Affiliations

  1. Department of Pharmaceutics and Drug Delivery, School of Pharmacy, The University of Mississippi, Oxford, Mississippi, USA

    Akshaya Tatke, Karthik Yadav Janga & Soumyajit Majumdar

  2. National Center for Natural Products Research, The University of Mississippi, Oxford, Mississippi, USA

    Bharathi Avula & Ikhlas A. Khan

  3. Department of Ophthalmology, Hamilton Eye Institute, The University of Tennessee Health Science Center, Memphis, Tennessee, USA

    XiangDi Wang & Monica M. Jablonski

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  1. Akshaya Tatke
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Correspondence to Soumyajit Majumdar.

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Tatke, A., Janga, K.Y., Avula, B. et al. P-glycoprotein Restricts Ocular Penetration of Loperamide across the Blood-Ocular Barriers: a Comparative Study in Mdr1a Knock-out and Wild Type Sprague Dawley Rats. AAPS PharmSciTech 19, 1662–1671 (2018). https://doi.org/10.1208/s12249-018-0979-2

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  • DOI: https://doi.org/10.1208/s12249-018-0979-2

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