中国科学院昆明动物研究所机构知识库

中风是全球范围的高发病率、高致死率、除血管问题外发病前无明显症状的突发性综合疾病。不管是对于中风的急性干预治疗，还是后期的恢复性治疗都存在一定的困难。因此，不断地发掘有效的抗中风药物以及对这些药物抗中风机制的研究，对于中风的预防、治疗及恢复都具有非常重要的意义。现今，世界多个国家都在尝试从植物中获取具有一定疾病治疗作用的小分子化合物。但只有很小一部分药用植物来源的化合物被发掘。例如，灯盏花、细辛、银杏叶和丹参等，这些中草药植物的提取物已应用于中风的中西临床联合治疗。 中国传统中药数据库记录了超过2000种的药用植物。这些药用植物被用于治疗中风已经有近千年的历史。由此可见，在有效的抗中风药物发掘方面，传统中草药无疑是一个宝贵资源库。然而，依靠实验系统地发掘传统中草药植物中有效地小分子化合物及其作用机制是一个庞大复杂且昂贵耗时的过程。 鉴于这一现状，本论文研究旨在结合中草药中所含的活性化合物小分子和已知的中风相关的作用靶点，利用分子对接的方法寻找中草药中能够对治疗中风起到关键作用的活性化合物，进而发掘中草药活性化合物在抗中风过程中的作用机制，明确其治疗中风的作用靶点，从而有效地预防和治疗中风。 首先，通过中国知网（CNKI）数据库，以“中风病”、“脑梗塞”和“脑出血”为检索主题，我们获得了5000多篇关于中风临床中草药治疗的研究。在这些相关的临床抗中风中草药的研究中，有801篇拥有完整的中药药方、用药人数和治疗效果等信息。进而，对每篇研究进行人工查阅和统计，获得954个临床抗中风中药药方、192个药方中所包含的抗中风中草药植物以及这些植物的使用频率。 其次，使用台湾中草药数据库TCM Database@Taiwan（录入8529个中草药植物ID和超过60000的中草药植物中所含的活性化合物小分子ID）中通过类药性筛选的21940个中草药活性小分子和汤森路透（Thomson Reuters）Integrity数据库中15个已知的中风相关靶点进行分子对接，从而获得2355个具有潜在作用于中风相关靶点能力的候选抗中风中草药化合物。 接着，将候选抗中风中草药化合物，通过TCM数据库获得其所对应的中草药植物中。结果显示，相对于随机选取的非抗中风中草药植物而言，候选化合物显著富集于使用频率前十的抗中风中草药植物。此外，抗中风靶点结构复合物上的配体小分子和Integrity数据库51个中风相关的药物，同候选化合物一起与靶点进行分子对接。分子对接能量值之间的对比表明，候选抗中风中草药活性化化合物具有更低的分子对接能量值，也更易于同抗中风靶点发生互作反应。这些发现都肯定了获得的2355个候选化合物潜在的抗中风作用。 然后，我们对候选抗中风中草药化合物进行同现有药物结构相似性分析、基于作用靶点的互作网络分析、基于共有分子指纹印记结构的聚类分析、与抗中风靶点的作用机制分析以及ADMET可药特性等方面的分析。 （1）1985个DrugBank上市药物中，16个药物化合物与19个候选抗中风中草药化合物结构上一致（Tc=1）。这些候选化合物和现有药物分别作用于不同的抗中风靶点。其中部分药物已用于抗中风的治疗研究。 （2）在2355个候选抗中风中草药化合物中，有1564个化合物单一靶向15个抗中风靶点；其中作用于抗中风靶点COX-1的候选化合物数目最多，有172个。791个化合物属于多靶点化合物，其作用的抗中风靶点数目从2到11不等；候选抗中风化合物18583麦角异胺（Ergotaminine）可作用于11个抗中风靶点。 （3）根据候选化合物的分子指纹印记特征，将候选化合物进行结构聚类。10个聚类簇的中心化合物分子都含有碳环结构，但每个簇的候选化合物成员数目不同，作用于每个靶点的候选化合物数目也不尽相同。 （4）临床抗中风中草药植物和其所含的候选抗中风化合物，在靶点NOS3、PSD-95和PDE5A上显著富集，表明这些抗中风靶点在中风治疗中具有重要作用。并进一步揭示了中草药植物和候选化合物的抗中风作用机制。 （5）经过ADMET成药性特征评价，候选抗中风中草药化合物中35个化合物有良好水溶性、肠吸收、血脑屏障渗透性、肝毒性和血浆蛋白结合力等特征。通过本论文的研究，发掘了天然中草药化合物中获得具有潜在抗中风活性的先导化合物及其作用的分子机制，对中风药物的开发设计有重要的指导性作用。

Stroke is a sudden comprehensive disease with globally high incidence and mortality, in addition to vascular problems, there is no obvious symptoms before onset. Whether for acute stroke intervention,or for the late restorative treatment, both have some difficulties. Therefore, it is of great significance for prevention, treatment and recovery of stroke to explore effective anti-stroke drugs and study their mechanism.Nowadays, many countries in the world are trying to get small molecule compounds with therapeutic effect on certain diseases from medicinal plants. But only a small fraction of the medicinal plant-derived compounds were explored. For example, Erigeron breviscapus, Asarum, Ginkgo biloba and Salvia miltiorrhiza and other herbal extracts of Chinese herbal medicine has been used in the combination of Chinese and Western clinical treatment of stroke. The traditional Chinese medicine database records more than 2000 medicinal plants. These medicinal plants have been used to treat stroke for nearly a thousand years. Thus, in the effective anti-stroke drug discovery, the traditional Chinese herbal medicine is undoubtedly a valuable resource. However, it is a complicated, expensive and time-consuming process to systematically explore the effective small molecule compounds and their mechanism in traditional Chinese herbal plants relying on experiments. In view of this situation, this paper aims to combine the small active compounds contained in Chinese herbal medicine and the known anti-stroke target, and use molecular docking method to find the key Chinese herbal medicine active compounds of anti-stroke, and then explore the mechanism of these active compounds and their targets in the process of stroke treatment, so as to effectively prevent and treat stroke.First, through the China National Knowledge Infrastructure(CNKI), we obtained more than 5 thousand stroke clinical Chinese medicine treatment researches with "stroke", "cerebral infarction" and "cerebral hemorrhage" as the searching theme. Of these related clinical trials, 801 have complete information of Chinese medicine prescriptions, the number of patients and treatment effects. In addition, manual statistics of each study were carried out to obtain 954 clinical anti-stroke Chinese medicine prescriptions, 192 anti-stroke plants contained from the prescription and the usage frequency of these anti-stroke plants for the clinical treatment of stroke.Second, 21940 Chinese active small molecules and Thomson Reuters 15 known anti-stroke targets undergo molecular docking so as to obtain 2355 candidate anti-stroke compound which has the potential to act with anti-stroke target. The former came from TCM Database@Taiwan (8529 Chinese plant ID and more than 60000 active compound ID contained from Chinese plants) by drug-based screening. The later obtained from Thomson Reuters Integrity database.Next, corresponding candidate anti-stroke compounds to anti-stroke plants according to TCM database. The results showed that the candidate compounds were significantly enriched in the top 10 anti-stroke plants, compared with the randomly selected 10 non-anti-stroke plants. In addition, ligands and the Integrity 51 stroke related drugs are used in molecular docking together with candidate anti-stroke compounds. The comparison of molecular docking energy values showed that candidate anti-stroke compounds had lower molecular docking energy values, and were more likely to react with anti-stroke target. These findings confirm the potential anti-stroke effect of 2355 candidate compounds.Then, we analyzed candidate anti-stroke compounds from similarity with the existing drug, interaction network with their anti-stroke targets, structure clustering based on their common molecular fingerprints, action mechanism with their anti-stroke targets, and the ADMET pharmacological properties.(1) Of the 1985 DrugBank marketed drugs, 16 drug compounds were structurally consistent with 19 candidate anti-stroke compounds (Tc=1). These candidate anti-stroke compounds and existing drugs act on different anti-stroke targets, respectively. Some of these drugs have been used in the treatment of stroke.(2) Among the 2355 candidate anti-stroke Chinese medicinal compounds, there are 1564 small molecules targeting a single target, and the number of candidate anti-stroke compounds acting on the target COX-1 is the largest, with a total of 172. The 791 candidate anti-stroke Chinese medicinal compounds belong to multiple target compounds, the target number of these compounds ranges from 2 to 11, and candidate anti-stroke compound 18583 (Ergotaminine) can acts with 11 anti-stroke targets.(3) According to the characteristics of molecular fingerprint for candidate anti-stroke compounds, the candidate anti-stroke compounds are clustered. Each of the 10 cluster center molecular contains carbocyclic structure, but the number of candidate anti-stroke compounds in each cluster is different, and the number of candidates compounds for their primary targets is not the same.(4) The significant enrichment of clinical anti-stroke Chinese medicinal plants and their candidate anti-stroke compounds at target NOS3, PSD-95 and PDE5A, indicating that these anti stroke targets play an important role in the treatment of stroke. And further revealed the anti-stroke mechanism of Chinese medicinal plants and their candidate anti-stroke compounds..(5) After the evaluation of the ADMET potability, 35 compounds of candidate anti-stroke Chinese herbal compounds had good water-solubility, intestinal absorption, blood-brain barrier permeability, hepatotoxicity and plasma protein binding. Through the research of this paper, the molecular mechanism of the lead compound with potential anti-stroke activity and its action in natural Chinese medicine compounds was explored, which has important guiding significance for the development and design of stroke drug.