作为一线临床药物，雷公藤苷片（TGTs）在疗效和毒副作用方面经常出现不一致的情况，主要是由于质量控制不足所致。因此，迫切需要临床相关的雷公藤苷片质量标准。基于化学物质并考虑药理学效应，我们旨在开发一种有效的雷公藤苷片质量评价方法。我们从不同制造商收集了代表性的商业样品，通过定性高效液相色谱-线性离子阱-轨道四极杆质谱联用（UHPLC/LTQ-Orbitrap-MS）和定量高效液相色谱串联质谱分析方法，成功地用于基于其化学性质评估其质量的相似性和差异性。然后，使用Jurkat、RAW264.7、MIA PaCa-2和PANC-1作为细胞模型，评估了雷公藤苷片及相关单体的抗免疫、抗炎和抗肿瘤活性。随后，我们通过分子对接来预测和验证小分子-DCTPP1的相互作用，使用建立的DCTPP1酶活性测定法。最后，我们进行了灰色关联分析，评估了通过不同制造商生产的雷公藤苷片的化学特性和生物效应。我们收集了来自5个制造商（公司A，公司B，公司C，公司D，公司E）的24批雷公藤苷片（D01-D24）进行质量评价。化学成分分析揭示了样品物质基础的显著差异。来自公司B的D02，D18-D20样品形成一个独立的分组，与其他样品有所不同，主要是由于三萜类化合物如雷公藤内酯、雷公藤酚内酯和雷公藤脱硫素在这些样品中的缺失。使用相同的雷公藤苷片浓度进行的体外抗免疫、抗肿瘤和抗炎试验显示，除了D02，D18-D20之外，其余的20个样品表现出不同程度的抗免疫、抗肿瘤和抗炎活性。我们的实验验证了三萜类化合物如雷公藤内酯、雷公藤酚内酯和雷公藤脱硫素均为DCTPP1的抑制剂，并且雷公藤苷片通常表现出DCTPP1的酶抑制活性。此外，来自公司B的D02，D18-D20样品的抑制活性远远低于其他样品，IC50差异接近10倍。进一步的综合分析揭示了这些样品的DCTPP1酶抑制活性与其抗免疫、抗肿瘤和抗炎活性之间的高度相关性。建立的DCTPP1酶活性测定法被证明非常适用于定量药理学和药学分析，以补充现有的雷公藤苷片质量控制体系并评估其有效性。© 2023 Elsevier GmbH版权所有。保留所有权利。

As first-line clinical drugs, tripterygium glycoside tablets (TGTs) often have inconsistent efficacy and toxic side effects, mainly due to inadequate quality control. Therefore, clinically relevant quality standards for TGTs are urgently required.Based on chemical substances and considering pharmacological efficacy, we aimed to develop an effective quality evaluation method for TGTs.Representative commercial samples of TGTs were collected from different manufacturers, and qualitative UHPLC/LTQ-Orbitrap-MS and quantitative UHPLC-MS/MS analysis methods were successfully applied to evaluate their quality similarities and differences based on their chemical properties. Then the anti-immunity, anti-inflammatory and antitumor activities of TGTs and related monomers were evaluated using Jurkat, RAW264.7, MIA PaCa-2, and PANC-1 as cellular models. Subsequently, we predicted and verified small molecule-DCTPP1 interactions via molecular docking using the established DCTPP1 enzymatic activity assay. Finally, we performed a gray relational analysis to evaluate the chemical characteristics and biological effects of TGTs produced by different manufacturers.We collected 24 batches of TGTs (D01-D24) from 5 manufacturers (Co. A, Co. B, Co. C, Co. D, Co. E) for quality evaluation. The chemical composition analysis revealed significant differences in the substance bases of the samples. The D02, D18-D20 samples from Co. B constituted a separate group that differed from other samples, mainly in their absence of diterpenoids and triterpenoids, including triptolide, triptophenolide, and triptonide. In vitro anti-immunity, antitumor and anti-inflammatory tests using the same TGT concentration revealed that, except for D02, D18-D20, the remaining 20 samples exhibited different degrees of anti-immunity, antitumor and anti-inflammatory activity. Our experiments verified that triptolide, triptophenolide, and triptonide were all DCTPP1 inhibitors, and that TGTs generally exhibited DCTPP1 enzyme inhibitory activity. Moreover, the inhibitory activity of D02, D18-D20 samples from Co. B was much lower than that of the other samples, with a nearly tenfold difference in IC50. Further comprehensive analysis revealed a high correlation between DCTPP1 enzyme inhibition activity and the anti-immunity and antitumor and anti-inflammatory activities of these samples.The established DCTPP1 enzymatic activity assay proved suitable for quantitative pharmacological and pharmaceutical analysis to complement the existing quality control system for TGTs and to evaluate their effectiveness.Copyright © 2023 Elsevier GmbH. All rights reserved.