Dose proportionality and bioavailability of quinoxaline-based JNK inhibitor after single oral and intravenous administration in rats.

The dose proportionality and bioavailability of the potential anti-inflammatory and neuroprotective JNK inhibitor 11H-indeno[1,2-b]quinoxalin-11-one oxime (IQ-1) were evaluated by comparing pharmacokinetic parameters after single oral (25, 50 and 100 mg/kg) and intravenous (1 mg/kg) IQ-1 administration in rats.IQ-1 and its major metabolite ketone 11H-indeno[1,2-b]quinoxalin-11-one (IQ-18) were isolated from plasma samples by liquid-liquid extraction. IQ-1 (E-isomer) and IQ-18 were simultaneously quantified in plasma by the validated method of liquid chromatography with triple quadrupole mass spectrometry (HPLC-MS/MS).The absolute bioavailability of IQ-1 was < 1.5%. Cmax values were 24.72 ± 4.30, 25.66 ± 7.11 and 37.61 ± 3.53 ng/mL after single oral administration of IQ-1 at doses of 25, 50 and 100 mg/kg, respectively. IQ-1 exhibited dose proportionality at 50-100 mg/kg dose levels, whereas its pharmacokinetics was not dose proportional over the range of 25-50 mg/kg. IQ-18 demonstrated the invariance of the dose-normalized Cmax.In this study we systematically elucidated the absorption characteristics of IQ-1 in rat gastrointestinal tract and provided better understanding of IQ-1 pharmacology for the future development of a new formulations and therapeutic optimisation.

Novel aspects of taxifolin pharmacokinetics: Dose proportionality, cumulative effect, metabolism, microemulsion dosage forms.

Taxifolin (TFL) is a small drug molecule with a broad therapeutic potential limited by its poor aqueous solubility and excessive metabolism. Despite comprehensive research, some aspects of the TFL pharmacokinetics, e.g., dose proportionality and possible cumulative effect, remain unexplored. In the current study, we have tried to fill this gap. Our results revealed that the TFL pharmacokinetics in rats had nonlinear character in the dose range of 10-50 mg/kg after its single oral administration (AUC). For Cmax, the data are ambiguous: linearity was confirmed via the equivalence criterion and was disproved using the power model approach. Also, the cumulative drug effect was observed on the 4th day after its multiple-dose oral administration (25 mg/kg; compared to the 1st day). Interestingly, biologically active TFL metabolites such as aromadendrin and luteolin were putatively found in plasma samples, although they were previously detected only in feces. In addition, oil-in-water and water-in-oil microemulsions were fabricated to design novel drug delivery systems. These carrier dosage forms did not improve the TFL bioavailability but significantly affected its metabolism. To support pharmacokinetic studies, the bioanalytical liquid chromatography-tandem mass spectrometry method was developed and validated in the concentration range of 1-1000 ng/mL using candesartan as an internal standard. Liquid-liquid extraction with methyl tert-butyl ether was used to isolate the analytes from plasma followed by evaporation and reconstitution of the residues in acetonitrile. Thus, the present findings broaden our understanding of the TFL behavior in vivo and provide novel ideas and reference directions for its continued use in medical practice.

LC-MS/MS method for the determination of a semi-synthetic phenolic antioxidant 2,6-diisobornyl-4-methylphenol in rats after different administration routes.

IBP (2,6-diisobornyl-4-methylphenol) is a small drug molecule with antioxidant properties considered to be a promising neuro-, cardio-, and retinoprotective agent. In this study, a bioanalytical LC-MS/MS method for its determination in rat plasma was developed using 11H-indeno[1,2-b]quinoxalin-11-one oxime as an internal standard (IS). The analytes were extracted from plasma by liquid-liquid extraction technique using isopropyl alcohol:chloroform mixture (1:5, v/v) followed by evaporation and reconstitution of the residues in acetonitrile. The chromatographic separation was carried out on the EC Nucleodur C8 ec column (150 × 4.6 mm, 5 µm) under an isocratic elution mode using acetonitrile and water containing 0.1% (v/v) formic acid (97:3, v/v) as a mobile phase at a flow rate of 0.55 mL/min (40 °C). The IS and IBP were eluted at 3.79 ± 0.02 and 6.30 ± 0.02 min, respectively. The total analysis time was 7.00 min. Multiple reaction monitoring was used to conduct the MS/MS detection in the negative ion mode with transitions at m/z 245.9 â†’ 214.9 (IS) and 379.2 â†’ 256.0 (IBP). Validation studies of the developed method revealed good linearity over the range of 10-5,000 ng/mL. Within- and between-run accuracy was in the range of 92-110%, while within- and between-run precision was below 8%. Additionally, low matrix effects and high recovery (above 98%) were observed. IBP remained stable in rat plasma at room temperature for 4 h, at -80 °C for 21 days, over three freeze-thaw cycles, under vacuum concentrator (45 °C, dried residues) and auto-sampler (15 °C, processed samples) temperatures for 1 h and 24 h, respectively. Subsequently, the validated LC-MS/MS method has been successfully applied to quantitate IBP in actual plasma samples after a single oral, intramuscular, and subcutaneous dose of IBP (10 mg/kg in the peach oil) to rats. Pharmacokinetic studies show that more rapid and complete IBP absorption with a satisfactory excretion rate were observed after oral administration route compared to the intramuscular and subcutaneous ones.

Quantification of a promising JNK inhibitor and nitrovasodilator IQ-1 and its major metabolite in rat plasma by LC-MS/MS.

Background: IQ-1 is a promising c-Jun-N-terminal kinase inhibitor and nitrovasodilator. An LC-MS/MS method was validated to determine IQ-1 isomers and major metabolite IQ-18 in rat plasma. Materials & methods: The analytes were extracted using ethyl acetate. The chromatographic separation was performed on a C8 column (150 × 4.6 mm, 5 µm) under acetonitrile-water (5 mM ammonium formate buffer, pH 2.93) gradient elution. Multiple reaction monitoring was used for MS/MS detection in the positive ion mode. Results: The method was fully validated over the range of 0.1-400 ng/ml (Z-isomer), 0.9-3600 ng/ml (E-isomer), 5.0-4000 (IQ-18). Conclusion: This method has been successfully applied to pharmacokinetic studies of IQ-1 and IQ-18 in rats after a single oral dose of IQ-1 (50 mg/kg).