ABSTRACT
The cassette dosing technique is employed in the drug discovery stage of non-clinical studies to obtain pharmacokinetic data from multiple drug candidates in a single experiment. The objective of the current investigation was to evaluate the effect of sex and food on the selected pharmacokinetic parameters of four biopharmaceutical classification system (BCS) drugs (BCS-I: propranolol, BCS-II: diclofenac, BCS-III: atenolol, and BCS-IV: acetazolamide) utilizing cassette dosing in male and female rats under fed and fasting conditions. Different animal groups were dosed intravenous (i.v) and oral at 1 and 10 mg/kg, respectively, in the form of cassette at a dose of 5 mL/kg. Blood samples were analyzed by liquid chromatography-tandem mass spectrometry. Pharmacokinetics parameters were calculated using Phoenix software version 8.1. A significant increase (p < 0.05) of the area under the plasma concentration-time (AUC0-last) was observed for diclofenac and acetazolamide in females over males after i.v dosing. Additionally, acetazolamide showed greater instantaneous concentration at the time of dosing, and clearance in females (p < 0.05) compared to males after i.v administration. After oral dosing, propranolol exhibited significant variations (p < 0.05) in the maximum drug concentration (Cmax), AUC0-last, the volume of distribution (Vd), and bioavailability in females as compared to males under fed state. Diclofenac showed significant changes (p < 0.05) in AUC0-last, and clearance (Cl) in females as compared to males under fasting and fed state. However, acetazolamide exhibited a significant enhancement (p < 0.05) in AUC0-last, Vd, and Cl in fasting females than the males. The data here illustrates that there is an appreciable difference in AUC and Cmax values exist in male and female rats under fed and fasting conditions administered with the cassette dosing of tested BCS class drugs.
Subject(s)
Biological Products , Pharmaceutical Preparations , Administration, Oral , Animals , Area Under Curve , Cross-Over Studies , Female , Male , RatsABSTRACT
These studies compared the efficacies of our clinical agent galeterone (Gal) and the FDA-approved prostate cancer drug, enzalutamide (ENZ) with two lead next generation galeterone analogs (NGGAs), VNPP414 and VNPP433-3ß, using prostate cancer (PC) in vitro and in vivo models. Antitumor activities of orally administered agents were also assessed in CWR22Rv1 tumor-bearing mice. We demonstrated that Gal and NGGAs degraded AR/AR-V7 and Mnk1/2; blocked cell cycle progression and proliferation of human PC cells; induced apoptosis; inhibited cell migration, invasion, and putative stem cell markers; and reversed the expression of epithelial-to-mesenchymal transition (EMT). In addition, Gal/NGGAs (alone or in combination) also inhibited the growth of ENZ-, docetaxel-, and mitoxantrone-resistant human PC cell lines. The NGGAs exhibited improved pharmacokinetic profiles over Gal in mice. Importantly, in vivo testing showed that VNPP433-3ß (at 7.53-fold lower equimolar dose than Gal) markedly suppressed (84% vs. Gal, 47%; p < 0.01) the growth of castration-resistant PC (CRPC) CWR22Rv1 xenograft tumors, with no apparent host toxicity. ENZ was ineffective in this CRPC xenograft model. In summary, our findings show that targeting AR/AR-V7 and Mnk1/2 for degradation represents an effective therapeutic strategy for PC/CRPC treatment and supports further development of VNPP433-3ß towards clinical investigation.
ABSTRACT
A rapid sensitive and selective MRM based method for the determination of polyethylene glycol 400 (PEG 400) in rat plasma was developed using liquid chromatography/tandem mass spectrometry (LC-MS/MS). PEG 400 and telmisartan (Internal standard) were extracted from rat plasma with acetonitrile and analysed on C18 column (Waters Xbridge, 50×4.6 mm, 3.5 µm) with the mobile phase (A - 0.1% formic acid in water; B - methanol). A generic gradient method with a short run time of 3.5 min was developed for the analysis of PEG 400. A total of nine oligomers were identified for PEG 400. The most abundant ions corresponding to PEG 400 oligomers at m/z 327, 371, 432, 476, 520, 564, 608, 652 and 696 with daughter ion at m/z 89 were selected for multiple reaction monitoring (MRM) in electrospray mode of ionisation. Analyte peak area of the oligomers was summed up to calculate the plasma concentrations of total PEG 400. The standard curve was linear (0.9954) over the concentration range of 1.01-1013.40 µg/mL. The lower limit of quantitation for PEG 400 was 1.01 µg/mL using 50 µL plasma. The coefficient of variation and relative error for inter and intraassay at three QC levels were 2.31-13.34 and -7.99 to 0.37, respectively. The method was validated for various parameters such as extraction recovery, matrix effect, autosampler stability, benchtop stability, freeze thaw stability, long term stability and was proved to be consistent across three QC levels with overall %CV less than 15. The developed method was successfully applied to the absolute bioavailability study of PEG 400 in male Sprague Dawley rats. Plasma concentrations of PEG 400 was measured after administration through oral and intravenous routes in male Sprague Dawley rats at a dose of 3.38 g/kg. Pharmacokinetic (PK) parameters were characterised by performing the analysis using Phoenix Winnonlin software (v 6.3). PEG 400 has good oral bioavailability with mean absolute bioavailability of 47.23%. Plasma concentration profile/PK parameters of PEG 400 was established in both intravenous and oral routes, which helps to qualify the analytical batch of NCEs having spiky plasma concentration profiles/erratic results. Purity of the PEG 400 oligomers was estimated using ELSD detection. Differences in pharmacokinetics of oligomers was studied. It was found that with increase in molecular weight of the oligomer, a decrease in absolute bioavailability was observed.
