Abstract
Celastrol (CL), a compound isolated from Tripterygium wilfordii, possesses various bioactivities such as antitumor, anti-inflammatory and anti-obesity effects. In previous studies, we developed CL-encapsulated silk fibroin nanoparticles (CL-SFNP) with satisfactory formulation properties and in vitro cancer cytotoxicity effect. For further in vivo oral bioavailability evaluation, in this study, a simple and reliable LC-MS/MS method was optimized and validated to determine CL concentration in rat plasma. The separation of CL was performed on a C18 column (150 by 2 mm, 5 µm) following sample preparation using liquid–liquid extraction with the optimized extraction solvent of tert-butyl methylether. The assay exhibited a good linearity in the concentration range of 0.5–500 ng/mL with the lower limit of quantification (LLOQ) of 0.5 ng/mL. The method was validated to meet the requirements for bioassay with accuracy of 91.1–110.0%, precision (RSD%) less than 9.1%, extraction recovery of 63.5–74.7% and matrix effect of 87.3–101.2%. The developed method was successfully applied to the oral bioavailability evaluation of CL-SFNP. The pharmacokinetic results indicated the AUC0-∞ values of CL were both significantly (p < 0.05) higher than those for pure CL after intravenous (IV) or oral (PO) administration of equivalent CL in rats. The oral absolute bioavailability (F, %) of CL significantly (p < 0.05) increased from 3.14% for pure CL to 7.56% for CL-SFNP after dosage normalization. This study provides valuable information for future CL product development.
Highlights
Celastrol (CL, Figure 1A), a quinone methide triterpenoid isolated from Tripterygium wilfordii, possesses various bioactivities such as antitumor, anti-inflammatory and anti-obesity effects, and have been demonstrated to have treatment potential on multiple cancers [1], autoimmune diseases [2], chronic inflammation [3], cardiovascular diseases [4] and neurodegenerative diseases [5]
In the LC-MS/MS method development, chromatographic condition was optimized, and the mobile phase with the additive of 0.1% formic acid-2 mM ammonium acetate in aqueous solution (A)
To mediate the contradiction between elution time and separation as well as to reduce matrix effect, we applied a gradient elution of 50% B at 0–2 min, 50–95% B at 2–3 min, 95% B at 3–6 min, 95–50% B at 6–7 min with the total analysis time of 10 min, and injected the eluent into the MS system at 5.5 min using the valve as a diverter
Summary
Celastrol (CL, Figure 1A), a quinone methide triterpenoid isolated from Tripterygium wilfordii, possesses various bioactivities such as antitumor, anti-inflammatory and anti-obesity effects, and have been demonstrated to have treatment potential on multiple cancers [1], autoimmune diseases [2], chronic inflammation [3], cardiovascular diseases [4] and neurodegenerative diseases [5]. The poor aqueous solubility, low therapeutic index and systemic toxicity of CL limited its clinical applications [3,6]. The clinical application of CL is limited mainly due to its unsatisfactory in vivo pharmacokinetic as well as pharmacodynamic profiles. In vitro drug release studies of CL-SFNP revealed a slow and sustained release of drug at physiological pH (pH 7.4) and rapid release at lysosomal pH (pH 4.5), and the in vitro cytotoxicity study against two human pancreatic cancer cell lines (PANC-1 and Mia PaCa-2) demonstrated increased growth inhibition with the nanoparticle formulation compared to pure CL. It is necessary to further study this formulation’s in vivo properties for possible product development of CL
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