Zhang Xiaojing, Liu Tong, Zhang Yidan, Liu Fanye, Li Haiying, Fang Dong, Wang Chaojie, Sun Hua, Xie Songqiang
School of Pharmacy, Institute for Innovative Drug Design and Evaluation, Henan University, Kaifeng, China.
The Key Laboratory of Natural Medicine and Immuno-Engineering, Henan University, Kaifeng, China.
Front Pharmacol. 2019 May 17;10:521. doi: 10.3389/fphar.2019.00521. eCollection 2019.
Cinobufotalin is one of the major anti-tumor components isolated from toad venom and has been used in the clinical therapy of hepatocellular carcinoma (HCC), known as Cinobufacini injection. However, the pharmacokinetic (PK) behaviors of cinobufotalin with HCC are still unknown. Hence, we have established a HCC model in Sprague Dawley (SD) rats induced by diethylnitrosamine (DEN), named as DEN-injured rats. Then, we developed and validated a sensitive and rapid ultra-performance liquid chromatography-tandem mass spectrometric (UPLC-MS/MS) method to quantify cinobufotalin in rat plasma. This UPLC-MS/MS method was successfully used to characterize the PK behaviors of cinobufotalin in normal and DEN-injured rats after intravenous (i.v.) injection at a dosage of 2.5 mg/kg. Cinobufotalin pharmacokinetics was well described by the two-compartment pharmacokinetic model and the PK parameters were calculated using WinNonlin 3.3 software. The transfer rate constant of cinobufotalin from the central compartment to the peripheral compartment (k) in DEN-injured rats was significantly greater than that in normal rats ( < 0.01), accompanied by the shorter half-life for the distribution phase (t). Additionally, the elimination rate constant (K) and clearance (CL) values in DEN-injured rats were significantly higher than that in normal rats ( < 0.05 for K and < 0.001 for CL, respectively). Therefore, the values of areas under concentration - time curve (AUC) and the liver concentration of cinobufotalin in DEN-injured rats was obviously lower than that in normal rats ( < 0.001 and < 0.01, respectively). This indicated that the PK behaviors of cinobufotalin will be altered in rats with HCC. In addition, P-glycoprotein (P-gp) has shown higher expression in live tissues of DEN-injured rats. Furthermore, cinobufotalin was identified as the substrate of P-gp using MDCK II and MDCK-MDR1 cell models for the first time. Consequently, P-gp will play an important role in the disposition of cinobufotalin , which provided a new combination therapy for the clinical treatment of HCC.
华蟾酥毒基是从蟾蜍毒液中分离出的主要抗肿瘤成分之一，已被用于肝细胞癌（HCC）的临床治疗，即华蟾素注射液。然而，华蟾酥毒基在HCC中的药代动力学（PK）行为仍不清楚。因此，我们在二乙基亚硝胺（DEN）诱导的Sprague Dawley（SD）大鼠中建立了HCC模型，命名为DEN损伤大鼠。然后，我们开发并验证了一种灵敏快速的超高效液相色谱-串联质谱（UPLC-MS/MS）方法，用于定量大鼠血浆中的华蟾酥毒基。该UPLC-MS/MS方法成功用于表征华蟾酥毒基在正常和DEN损伤大鼠静脉注射（i.v.）2.5 mg/kg剂量后的PK行为。华蟾酥毒基的药代动力学通过二室药代动力学模型得到很好的描述，并使用WinNonlin 3.3软件计算PK参数。DEN损伤大鼠中华蟾酥毒基从中央室向外周室的转运速率常数（k）显著大于正常大鼠（<0.01），分布相半衰期（t）较短。此外，DEN损伤大鼠的消除速率常数（K）和清除率（CL）值显著高于正常大鼠（K<0.05，CL<0.001）。因此，DEN损伤大鼠中华蟾酥毒基的浓度-时间曲线下面积（AUC）值和肝脏浓度明显低于正常大鼠（分别为<0.001和<0.01）。这表明HCC大鼠中华蟾酥毒基的PK行为会发生改变。此外，P-糖蛋白（P-gp）在DEN损伤大鼠的肝脏组织中表达较高。此外，首次使用MDCK II和MDCK-MDR1细胞模型确定华蟾酥毒基为P-gp的底物。因此，P-gp在华蟾酥毒基的处置中起重要作用，这为HCC的临床治疗提供了一种新的联合治疗方法。
