Background:Salviae Miltiorrhizae Radix et Rhizoma (Red Sage root) is widely usedin traditional Chinese medicine (TCM) for the treatment of Alzheimer’s disease (AD) withdemonstrated curative effects, based on the concept of "one drug with multiple therapeutic targets,"which appears to be a good strategy for AD treatment.Objective:This study aimed to develop of high-throughput screening (HTS) method for multitherapeutictarget components found in complex TCMs, which are active against AD, using RedSage root as the case study.Method:Acetylcholinesterase (AChE) inhibitors (AChEIs) from Red Sage root extracts werepre-screened by ultrafiltration-HPLC (UF-HPLC) analysis, in which AChE was added to the extractand then ultrafiltered to remove non-binding compounds. Potential AChEIs were identifiedby HPLC analysis of compounds bound to AChE. A microplate-based HTS was then used toquantify the AChE inhibitory activity and antioxidant activity of the pre-screened compounds.Results:Pre-screening found ten potential inhibitors, which were identified by ESI-TOF/MS; sixof these were purified by semi-preparative HPLC: Oleoyl neocryptotanshinone (1), DihydrotanshinoneⅠ (2), Cryptotanshinone (3), Tanshinone Ⅰ (4), Tanshinone ⅡA (5) and Miltirone (6).All six compounds had good AChE inhibitory activity and weak DPPH scavenging capacity.Conclusion:This study provides a platform and technology support for the rapid discovery ofmulti-target components, potentially active against AD, from complex TCMs and with strong potentialfor adaptation to the discovery of treatments for other diseases.

中文翻译：

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UHPLC结合高通量筛选的中药治疗阿尔茨海默病多靶点先导化合物快速筛选的综合策略——以丹参为例

背景：丹参（红鼠尾草根）在中医药（TCM）中广泛用于治疗阿尔茨海默病（AD），基于“一药多靶”的理念，疗效显着。目的：本研究旨在以红鼠尾草根为案例研究，开发对复杂中药中对AD有活性的多治疗靶点成分的高通量筛选（HTS）方法。方法：乙酰胆碱酯酶来自红鼠尾草根提取物的 (AChE) 抑制剂 (AChEI) 通过超滤-HPLC (UF-HPLC) 分析进行预筛选，其中将 AChE 添加到提取物中，然后超滤以去除非结合化合物。通过对与 AChE 结合的化合物的 HPLC 分析鉴定潜在的 AChEI。然后使用微孔板HTS对预筛选化合物的AChE抑制活性和抗氧化活性进行定量。结果：预筛选发现10个潜在抑制剂，通过ESI-TOF/MS鉴定；其中六种通过半制备型HPLC纯化：油酰新隐丹参酮（1）、二氢丹参酮Ⅰ（2）、隐丹参酮（3）、丹参酮Ⅰ（4）、丹参酮ⅡA（5）和米替隆（6）。六种化合物均具有良好的AChE抑制作用活性和较弱的 DPPH 清除能力。结论：本研究为从复杂的中药中快速发现具有潜在抗 AD 活性的多靶点成分提供了平台和技术支持，并具有很大的适应其他疾病治疗方法的潜力。