Circumvention of P-gp and MRP2 mediated efflux of lopinavir by a histidine based dipeptide prodrug.

Abstract

PURPOSE:

This study was aimed to develop a novel Histidine-Leucine-Lopinavir (His-Leu-LPV) dipeptide prodrug and evaluate its potential for circumvention of P-gp and MRP2-mediated efflux of lopinavir (LPV) indicated for HIV-1 infection.

METHODS:

His-Leu-LPV was synthesized following esterification of hydroxyl group of LPV and was identified by (1)H NMR and LCMS/MS techniques. Aqueous solubility, stability and cell cytotoxicity of prodrug was determined. Uptake and permeability studies were carried out using P-gp (MDCK-MDR1) and MRP2 (MDCK-MRP2) transfected cell lines. To further delineate prodrug uptake, prodrug interaction with influx transporters (PepT1 and PHT1) was determined. Enzymatic hydrolysis and reconversion of His-Leu-LPV to LPV was examined using Caco-2 cell homogenates.

RESULTS:

Aqueous solubility generated by the prodrug was markedly higher relative to unmodified LPV. Importantly, His-Leu-LPV displayed significantly lower affinity towards P-gp and MRP2 as evident from higher uptake and transport rates. [3H]-GlySar and [3H]-l-His uptake receded to approximately 30% in the presence of His-Leu-LPV supporting the PepT1/PHT1 mediated uptake process. A steady regeneration of LPV and Leu-LPV in Caco-2 cell homogenates indicated His-Leu-LPV undergoes both esterase and peptidase-mediated hydrolysis.

CONCLUSION:

Histidine based dipeptide prodrug approach can be an alternative strategy to improve LPV absorption across poorly permeable intestinal barrier.

• Authors: Abhirup Mandal, Dhananjay Pal, Ashim K Mitra
• Journal: International journal of pharmaceutics (Int J Pharm) Vol. 512 Issue 1 Pg. 49-60 (Oct 15 2016) ISSN: 1873-3476 [Electronic] Netherlands
• PMID: 27543355 (Publication Type: Journal Article)
• Copyright: Copyright © 2016 Elsevier B.V. All rights reserved.

Chemical References

• ABCC2 protein, human
• ATP Binding Cassette Transporter, Subfamily B, Member 1
• Carrier Proteins
• Dipeptides
• Membrane Transport Proteins
• Multidrug Resistance-Associated Protein 2
• Multidrug Resistance-Associated Proteins
• Nerve Tissue Proteins
• Peptide Transporter 1
• Prodrugs
• SLC15A4 protein, human
• Symporters
• Lopinavir
• histidylleucine

Topics

• ATP Binding Cassette Transporter, Subfamily B, Member 1 (metabolism)
• Carrier Proteins (metabolism)
• Cell Survival (drug effects)
• Cells, Cultured
• Dipeptides (chemistry, pharmacokinetics)
• Drug Stability
• Humans
• Lopinavir (chemistry, pharmacokinetics, pharmacology)
• Membrane Transport Proteins
• Multidrug Resistance-Associated Protein 2
• Multidrug Resistance-Associated Proteins (metabolism)
• Nerve Tissue Proteins (metabolism)
• Peptide Transporter 1
• Prodrugs (chemical synthesis, chemistry, pharmacokinetics, pharmacology)
• Solubility
• Symporters (metabolism)
• Transfection