Determination and pharmacokinetic study of the diacid metabolite of norcantharidin in beagle plasma by use of liquid chromatography–tandem mass spectrometry

Abstract

Because norcantharidin (NCTD) is unstable and subject to ring opening and hydrolysis, the diacid metabolite of norcantharidin (DM-NCTD) is the stable form of NCTD found in normal saline solution. Conversion of NCTD to DM-NCTD is almost 100%, making it possible to determine and investigate the pharmacokinetics of DM-NCTD converted from NCTD. In this paper, a sensitive, simple and selective liquid chromatographic-tandem mass spectrometric method was developed and validated for determination of DM-NCTD in beagle plasma. DM-NCTD was detected in multiple-reaction monitoring (MRM) mode by using the dehydrated ion 169.3 as precursor ion and its product ion 123.1 as the detected ion. Ribavirin was used as internal standard and detected in MRM mode by use of precursor ions, resulting in a product ion transition of m/z 267.1 → 135.1. This method was successfully used for a pharmacokinetic study of DM-NCTD in beagles after intravenous administration of DM-NCTD in normal saline solution at doses of 0.39, 0.78, and 1.6 mg kg(-1). DM-NCTD had dose-dependent kinetics across the dosage range investigated, with enhanced T(1/2α) and AUC(0-12) and apparently decreasing V(d) and CL with increasing dosage. After single-dose administration, T(1/2α) ranged from 0.20 to 0.55 h, AUC(0-12) from 1.81 to 43.6 μg mL(-1) h(-1), V(d) from 228 to 55.9 mL kg(-1), and CL from 220 to 36.5 mL kg(-1) h(-1) (P < 0.01). The results indicated nonlinear pharmacokinetic behavior of DM-NCTD in beagles, suggesting that the risk of DM-NCTD in normal saline solution intoxication may be non-proportionally increased at higher doses.

Folate receptor-targeted liposomes loaded with a diacid metabolite of norcantharidin enhance antitumor potency for H22 hepatocellular carcinoma both in vitro and in vivo

“…9 The pharmacokinetic study revealed that the risk of DM-NCTD intoxication may increase at high doses, and these results were consistent with the reported adverse effects for adults administered the high clinical dosage in the PRC. It is precisely because of these reasons that our study focused on increasing the t ½ and decreasing the potential adverse effects of DM-NCTD when the drug was used at a moderate dose.…”

“…The apparatus and LC-MS/MS conditions were the same as those described in our previous study. 9 Whole tissues and tumor samples were collected from the mice and stored at −80°C until use. The samples were homogenized in saline (tissue or tumor:water ratio of 1:4, w/v).…”

“…In a previous study, we confirmed the prolonged circulation characteristics of related parameters and increased the relative bioavailability (F r ) of DM-NCTD liposomes (compared with DM-NCTD as the reference formulation) in Kunming mice at a moderate dose (2 mg/kg) converted from the clinical dosage used in the People's Republic of China (PRC). 9,38 Although the results indicated that a liposomal drug-delivery system could have the potential to overcome the shortcomings of DM-NCTD by improving F r and increasing therapeutic efficacy, further in vitro and in vivo studies on tumor-targeted FR-mediated DM-NCTD liposome therapy are still needed to confirm these results. Therefore, the aim of this study was to develop tumor-targeted delivery for DM-NCTD and confirm its targeting characteristics at a moderate dose.…”

Effect of norcantharidin on the proliferation, apoptosis, and cell cycle of human mesangial cells

“…NCTD was previously shown to induce cell-cycle arrest through G2 arrest. 21 , 22 However, some groups have also suggested that NCTD inhibits the cell cycle at the G1 or the S phase. 8 , 23 As we know, there were two important stages in the cell cycle, which were the G2 phase to the M phase and the G1 phase to the S phase.…”

Review targeted drug delivery systems for norcantharidin in cancer therapy

“…Although NCTD greatly reduces the toxicity of CTD, there is still a certain degree of urinary toxicity, and organ toxicity in high-dose or long-term use [ 42 , 43 ]. Moreover, the poor solubility, short half-life, fast metabolism, as well as high venous irritation and weak tumor targeting ability limit its wide clinical application [ 44 – 46 ].…”