- 作者：Abhirup Mandal ; Dhananjay Pal ; Ashim K. Mitra ; Ph.D. ; mitraa@umkc.edu" class="auth_mail" title="E-mail the corresponding author (Professor of Pharmacy Chair)
- 关键词：HAART ; Highly active antiretroviral therapy ; HIV ; Human immunodeficiency virus ; LPV ; Lopinavir ; PHT1 ; Peptide/histidine transporter ; PepT1/PepT2 ; Peptide transporters ; TEER ; Transepithelial electric resistance ; GlySar ; Glycylsarcosine ; His ; Histidine ; Leu ; Leucine ; P-gp ; Permeability glycoprotein ; MRP2 ; Multidrug resistance-associated protein 2
- 刊名：International Journal of Pharmaceutics
- 出版年：2016
- 出版时间：15 October 2016
- 年：2016
- 卷：512
- 期：1
- 页码：49-60
- 全文大小：3287 K
Methods
His-Leu-LPV was synthesized following esterification of hydroxyl group of LPV and was identified by 1H 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.