Kinetic release of triptolide after injection of renal‐targeting 14‐succinyl triptolide‐lysozyme in a rat kidney study by liquid chromatography/mass spectrometry

Abstract

Triptolide (TP) is one of the most important biologically active components of the Chinese herb Tripterygium wilfordii Hook f (TWHf). A novel high-performance liquid chromatography/mass spectrometry (LC/MS) method was developed to study the kinetic release of TP after intravenous injection of the newly synthesized 14-succinyl triptolide-lysozyme (TPS-LZM). The method was validated in terms of selectivity, linearity, precision, accuracy, stability, limit of detection (LOD) and limit of quantification (LOQ). The calibration curve was linear over the concentration range 0.5-400.0 ng/g. The mean recovery of triptolide from spiked samples, in a concentration range of 0.5-400.0 ng/g, was 91.84% (RSD = 3.69%, n = 3). The intra- and inter-assay coefficients of variation were less than 6.38%. The method with simple sample pretreatment and being highly specific and precise, can be used for analysis of triptolide release in rat kidney after intravenous injection of renal-targeting TPS-LZM conjugate. The results showed that, as compared with free TP, TPS-LZM could significantly increase the concentration and prolong the action time of TP in the kidney.

The targeting of 14-succinate triptolide-lysozyme conjugate to proximal renal tubular epithelial cells

“…Endogenous low molecular weight proteins (LMWP), lysozyme, was utilized as renal-specific drug carriers to form 14-succinate triptolide-lysozyme (TPS-LZM) conjugate [14][15][16]. Our data demonstrated the efficacy and safety of this PTEC specific TP therapeutics as well as its potential clinical applications as a local immunosuppressant.…”

“…These mixtures were hydrolyzed at room temperature for 20 min. TP concentration was then analyzed by HPLC/MS [15].…”

“…TPS-LZM was added to the incubation medium at the start of the uptake experiment, and uptake was terminated after 3 min of incubation (unless otherwise specified) by the addition of 1 mL of ice-cold buffer followed by immediate aspiration. The monolayers were rinsed twice with ice-cold buffer and digested with 1 mL of 1 M NaOH, neutralized by HCl, and then analyzed for TP [15]. Protein contents of cell digests were estimated on parallel wells, using the Pierce BCA protein assay reagent kit (Pierce, USA), with bovine serum albumin as the standard.…”

Traditional Chinese Medicine Integrated Responsive Microneedles for Systemic Sclerosis Treatment

“…By employing a template-assisted layer-by-layer curing method, such MNs with triptolide (TP)/paeoniflorin (Pae) needle tips and BP-hydrogel needle bottoms were generated. TP, as the main component of T. wilfordii , has excellent anti-inflammatory, detoxification, and immunomodulatory effects [ 9 , 42 , 43 ], while Pae is an important extractive of P. lactiflora , which can further enhance the therapeutic effect and decrease the toxicity of TP [ 44 – 46 ]. When the traditional Chinese medicine integrated responsive MNs were pasted on the skin, their mechanical properties could bring about instant skin penetration to fully contact the lesion regions.…”