School of Chinese Materia Medica, Tianjin University of Traditional Chinese Medicine, Nankai District, Tianjin 300193, PR China.
School of Chinese Materia Medica, Tianjin University of Traditional Chinese Medicine, Nankai District, Tianjin 300193, PR China; Second Affiliated Hospital of Tianjin University of TCM, Tianjin 300150, PR China.
Phytomedicine. 2019 Feb 15;54:357-364. doi: 10.1016/j.phymed.2018.03.031. Epub 2018 Mar 19.
The quality standard of Tripterygium glycosides tablet (TGT) by CFDA can not fully reflect the effectiveness and safety. While, Q-marker was proposed to solve the problem of traditional Chinese medicine. PK-marker is mainly used to reflect the material exposure and the influencing factors of Chinese medicine after administration.
Based on the study of quantitative analysis, cytotoxicity and pharmacokinetics, this study screened out and confirmed whether wilforine could be served as a potential Q-marker and PK-marker of TGT.
A sensitive and selective UPLC-MS/MS method was developed and applied to quantitative research of TGT preparation and pharmacokinetics study of TGT. Then, HepG2 cells assay was used to evaluate the cytotoxicity induced by alkaloids in TGT. Then, a PK-PD research was carried out in adjuvant arthritis (AA) rats and control rats after oral administration of TGT, with different dosage and timing. The pharmacokinetic characteristics were determined and calculated by DAS1.0. The pharmacodynamics of TGT was evaluated by the change of paw swelling through one-way ANOVA analysis.
The quality of four alkaloids showed significant difference among four manufacturers, and they were abundant component in TGT from three manufacturers of all. HepG2 cells test revealed that wilforine and wilforgine could induce the cytotoxicity obviously. Pharmacodynamics index suggested that TGT had therapeutic effect on adjuvant arthritis. Thus, the four cases of death occurred in the high dose AA rat group had proven the significant toxicity caused by continuous high dose TGT administration. Furthermore, the result of pharmacokinetic study proved that C, and AUC of wilforine have dose-dependent and time-dependent characteristics. But for wilforgine, there was no indication that there was an accumulation phenomenon in vivo and its plasma concentration showed low exposure. Therefore, it could hardly become the PK-marker of TGT.
Wilforine is proposed as a biologically active and toxic component of TGT that can be served both as Q-marker and PK-marker. The quality, clinical safety, and efficacy of TGT should be evaluated by the quality of wilforine.
国家食品药品监督管理总局（CFDA）制定的雷公藤多苷片质量标准不能充分反映其有效性和安全性。而 Q 标志物被提出以解决中药的问题。PK 标志物主要用于反映给药后中药的物质暴露和影响因素。
本研究通过定量分析、细胞毒性和药代动力学研究，筛选并确定钩吻碱是否可作为雷公藤多苷片的潜在 Q 标志物和 PK 标志物。
建立并应用灵敏、选择性的 UPLC-MS/MS 方法对雷公藤多苷片进行定量研究和药代动力学研究。然后，采用 HepG2 细胞试验评估雷公藤多苷片中生物碱的细胞毒性。然后，在佐剂性关节炎（AA）大鼠和对照大鼠中进行 PK-PD 研究，分别给予不同剂量和时间的雷公藤多苷片。采用 DAS1.0 确定和计算药代动力学特征。通过单向方差分析评估雷公藤多苷的药效学。
四个生物碱的质量在四个厂家之间存在显著差异，并且它们在三个厂家的雷公藤多苷片中都是丰富的成分。HepG2 细胞试验表明，钩吻碱和钩吻宁可明显诱导细胞毒性。药效学指标表明，雷公藤多苷对佐剂性关节炎有治疗作用。因此，高剂量 AA 大鼠组的四起死亡病例证实了连续高剂量雷公藤多苷给药引起的显著毒性。此外，药代动力学研究结果表明，钩吻碱的 Cmax 和 AUC 具有剂量依赖性和时间依赖性特征。但是对于钩吻宁，体内没有表明存在蓄积现象，其血浆浓度显示出低暴露。因此，它很难成为 TGT 的 PK 标志物。
钩吻碱被提议为雷公藤多苷的生物活性和毒性成分，可同时作为 Q 标志物和 PK 标志物。雷公藤多苷的质量、临床安全性和疗效应通过钩吻碱的质量来评估。
