Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China; Second Affiliated Hospital of Tianjin University of Traditional Chinese Medicine, Hebei District, Tianjin 300150, China.
Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China.
Phytomedicine. 2018 May 15;44:155-163. doi: 10.1016/j.phymed.2018.02.006. Epub 2018 Feb 21.
Diabetes is a chronic disease associated with significant morbidity and mortality. Tangzhiqing tablet (TZQ), a Chinese traditional patent medicine, has been in phase 2 clinical trial for the treatment of diabetes mellitus. However, the current quality evaluation of TZQ still remains rather obscure.
The promising quality markers (Q-markers) of TZQ will be sought for its quality assessment and process control based on the qualitative, quantitative and dose-exposure-response analysis.
The fingerprint analysis of TZQ was carried out through ultra high performance liquid chromatography- quadrupole time-of-flight/ mass spectrometry (UPLC-Q-TOF/MS) assay. Multicomponent quantitative analysis was implemented to the main ingredients of nuciferine, paeoniflorin, salvianolic acid B, hyperoside and rutin in TZQ by means of LC analysis. The dose-exposure-response relationship of TZQ was revealed by a placebo-controlled, 5-way crossover study in healthy Chinese subjects. The potential Q-markers in plasma were determined by a liquid chromatography combined with tandem mass spectrometry (LC-MS/MS) method. The therapeutic effect of TZQ was expressed as the glucose-lowering profile. The exposure-response relevance was developed between the concentration of Q-markers and the glucose-lowering effect of TZQ.
46 compounds were identified or tentatively characterized from TZQ. The contents of paeoniflorin, nuciferine, salvianolic acid B, hyperoside and rutin in TZQ were 6.40, 1.75, 1.70, 0.004, and 0.006 mg, respectively. However, salvianolic acid B, hyperoside and rutin could hardly be detected in human plasma under the current LC-MS/MS condition. The exposures of nuciferine and paeoniflorin (AUC, C) were dose-proportionality in human at the studied dosage ranges. The glucose-lowering effect appeared to increase proportionally with increasing TZQ dose in healthy volunteers. A clockwise hysteresis was displayed between the exposure of nuciferine and paeoniflorin and the glucose-lowering effect of TZQ.
Nuciferine and paeoniflorin were identified as the promising Q-markers of TZQ based on the fingerprint qualitative analysis, multicomponent quantitative analysis and dose-exposure-response analysis. The two Q-markers are meaningful to ensure the quality assessment and process control of TZQ.
糖尿病是一种与显著发病率和死亡率相关的慢性疾病。糖脂清片（TZQ）是一种中药传统专利药，已进入 2 期临床试验，用于治疗糖尿病。然而，目前 TZQ 的质量评价仍然相当模糊。
基于定性、定量和剂量-暴露-反应分析，寻找 TZQ 的有前途的质量标志物（Q 标志物），用于其质量评估和过程控制。
采用超高效液相色谱-四极杆飞行时间/质谱联用（UPLC-Q-TOF/MS）法对 TZQ 进行指纹图谱分析。采用 LC 分析对 TZQ 中的主要成分荷叶碱、芍药苷、丹酚酸 B、金丝桃苷和芦丁进行了多组分定量分析。通过在中国健康受试者中进行的安慰剂对照、5 向交叉研究，揭示了 TZQ 的剂量-暴露-反应关系。通过液相色谱-串联质谱（LC-MS/MS）法确定了潜在的血浆 Q 标志物。TZQ 的治疗效果表示为降低血糖的概况。在 Q 标志物的浓度与 TZQ 的降血糖作用之间建立了暴露-反应相关性。
从 TZQ 中鉴定或初步鉴定了 46 种化合物。TZQ 中芍药苷、荷叶碱、丹酚酸 B、金丝桃苷和芦丁的含量分别为 6.40、1.75、1.70、0.004 和 0.006mg。然而，在当前的 LC-MS/MS 条件下， salvianolic acid B、hyperoside 和 rutin 几乎无法在人血浆中检测到。在所研究的剂量范围内，人血浆中荷叶碱和芍药苷（AUC、C）的暴露量呈剂量比例关系。健康志愿者中，随着 TZQ 剂量的增加，降糖效果呈比例增加。在荷叶碱和芍药苷的暴露量与 TZQ 的降糖效果之间显示出顺时针滞后。
基于指纹定性分析、多组分定量分析和剂量-暴露-反应分析，鉴定荷叶碱和芍药苷为 TZQ 的有前途的 Q 标志物。这两个 Q 标志物对于确保 TZQ 的质量评估和过程控制具有重要意义。
