The simultaneous determination of naringenin and its valine carbamate prodrug in rat plasma using supercritical fluid chromatography -tandem mass spectrometric method.

To enhance the oral bioavailability of naringenin, a valine carbamate prodrug was firstly synthesized. It is essential to measure naringenin and its carbamate prodrug simultaneously for evaluating their pharmacokinetic behavior in Sprague-Dawley rats. Here, the samples were analyzed by a supercritical fluid chromatography-tandem mass spectrometric (SFC-MS/MS) method after extracting by liquid-liquid extraction with ethyl acetate. The analytes were eluted completely on an ACQUITY UPC2TM BEH 2-EP column (3.0 × 100 mm, 1.7 µm) within 2.5 min by gradient elution. The mass transition ion pairs were m/z 273.2→153.0, 416.0→153.1, and 271.2→91.0 for naringenin, the prodrug, and genistein (the internal standard), respectively. Naringenin and the prodrug had excellent linear correlations over the range of 2-1000 ng/mL (r > 0.995) and 4-2000 ng/mL (r > 0.998), with lower limits of quantification of 2 ng/mL and 4 ng/mL, respectively. The intra-day and inter-day precision and accuracy for all quality control samples were within ± 15 %. The high-throughput, sensitive, and economical SFC-MS/MS method was successfully applied to the pharmacokinetic study of naringenin and its carbamate prodrug for the first time. The pharmacokinetic study results showed the total Cmax of naringenin in prodrug group was 4.14-fold higher than naringenin group. The higher total AUC value observed with prodrug group indicated increased bioavailability of naringenin as compared to naringenin suspension. The present work provides some helpful information for future studies of naringenin and its carbamate prodrug.

Bioactivatable Pseudotripeptidization of Cyclic Dipeptides To Increase the Affinity toward Oligopeptide Transporter 1 for Enhanced Oral Absorption: An Application to Cyclo(l-Hyp-l-Ser) (JBP485).

The cyclic dipeptides generally present lower affinity toward intestinal oligopeptide transporter 1 (PEPT1) than the linear dipeptides. JBP485 (cyclo(l-Hyp-l-Ser)) is a low-affinity substrate of PEPT1 with poor oral bioavailability. However, JBP923 (l-Hyp-l-Ser) is a high-affinity substrate of PEPT1 with high oral absorption. We hypothesize that the bioactivatable pseudo-tripeptidization prodrug strategy is promising to increase the affinity of cyclic dipeptides toward PEPT1. To test our hypothesis, we design five amino acid ester prodrugs of JBP485. Compared with JBP485, the optimal prodrug (JBP485-3-CH2-O-valine, J3V) demonstrates improved affinity of PEPT1, oral bioavailability in rats and beagle dogs. Moreover, J3V can dose-dependently protect against liver injury. Additionally, J3V is stable in the gastrointestinal tract, beneficial to the PEPT1-mediated membrane transport, and is bioactivated in the enterocytes and hepatic cells, essential to elicit its bioactivity. In summary, the bioactivatable pseudo-tripeptidization strategy shows potential in increasing affinity of PEPT1 to enhance oral bioavailability of cyclic dipeptides.