College of Pharmaceutical Science, Zhejiang University of Technology, No. 18, Chaowang Road, Hangzhou, 310014, China.
State Key Laboratory of Generic Manufacture Technology of Chinese Traditional Medicine, Shandong, 276006, China; Lunan Pharmaceutical Group Co., Ltd., Shandong, 276006, China.
J Ethnopharmacol. 2021 Jan 10;264:113282. doi: 10.1016/j.jep.2020.113282. Epub 2020 Sep 3.
Shenqi Jiangtang granule (SJG) is an ancient Chinese herbal formula used for treatment of Diabetes mellitus and its complications.
To establish an integrated approach for discovery of effective Aldose reductase inhibitors (ARIs) from SJG.
An integrated approach combining ultrafiltration-liquid chromatography-mass spectrometry (UF-LC-MS) with in silico molecular docking was established for development of ARIs. AR enzyme was separated from the rabbit's crystalline lens. The inhibitory activities of these compounds were detected by UV spectrophotometry with DL-glyceraldehyde as a substrate. Furthermore, molecular docking was used to understand the binding mechanism of these screened compounds interacting with AR.
After optimization of AR reaction system and ultrafiltration incubation system, 17 active ingredients were screened from SJG by UF-LC-MS technique. Among these potential AR inhibitors, ginsenoside Rd exhibited the strongest activity with IC value of 45.77 μM. Three of them, calycosin, gomisin J and schisandrin A were demonstrated to be potential inhibitors for the first time, with IC at 447.34 μM, 181.73 μM, and 429.00 μM, respectively. Most of the active compounds exhibited competitive inhibition against AR. The docking scores of saponins were higher than that of lignans, which was consistent with the verification results.
The results indicated that TCM formula with clinical efficacy was indeed hopeful source for screening active ingredients, and the combination of UF-LC-MS and in silico molecular docking was a universal and promising approach for development of effective enzyme inhibitors.
参芪降糖颗粒（SJG）是一种古老的中药配方，用于治疗糖尿病及其并发症。
建立一种从 SJG 中发现有效的醛糖还原酶抑制剂（ARIs）的综合方法。
建立了一种结合超滤-液相色谱-质谱联用（UF-LC-MS）和计算机分子对接的综合方法，用于开发 ARIs。从兔晶状体中分离出 AR 酶。用紫外分光光度法以 DL-甘油醛为底物检测这些化合物的抑制活性。此外，还使用分子对接来了解这些筛选化合物与 AR 相互作用的结合机制。
在优化 AR 反应体系和超滤孵育体系后，通过 UF-LC-MS 技术从 SJG 中筛选出 17 种活性成分。在这些潜在的 AR 抑制剂中，人参皂苷 Rd 表现出最强的活性，IC 值为 45.77 μM。其中 3 种，毛蕊异黄酮、戈米辛 J 和五味子 A，首次被证明是潜在的抑制剂，IC 值分别为 447.34 μM、181.73 μM 和 429.00 μM。大多数活性化合物对 AR 表现出竞争性抑制作用。皂苷的对接分数高于木脂素，这与验证结果一致。
这些结果表明，具有临床疗效的中药配方确实是筛选有效成分的有希望的来源，而 UF-LC-MS 与计算机分子对接的结合是一种通用且有前途的开发有效酶抑制剂的方法。
