基于化学信息学的抗心衰相关药物筛选和作用机制探讨

发布时间：2018-06-28 20:56

本文选题：化学信息学 + 心衰　；参考：《第二军医大学》2017年博士论文

【摘要】：心衰是一种常见的、昂贵的、可能致命的疾病,近年来,心衰的患病率和发病率正在逐年增加,心衰疾病已经发展成为一个常见的公共问题。本论文采用化学信息学手段虚拟筛选靶向心衰潜在靶标蛋白肿瘤坏死因子-α的小分子拮抗剂,并进行实验验证。同时,将化学信息学工具与网络药理学理念相整合,探讨四逆汤抗心衰的潜在作用机制,并构建了一个心衰相关中草药网络药理平台,为抗心衰中药复方、药材和化合物的靶标鉴定及网络药理机制探讨提供了一个专有的网络平台。1.基于虚拟筛选的肿瘤坏死因子-α小分子拮抗剂的预测和验证肿瘤坏死因子-α是治疗心衰疾病的潜在靶标蛋白,然而,目前报道的肿瘤坏死因子-α小分子抑制剂数量非常有限。本研究拟将虚拟筛选与生物验证相结合,快速高效地筛选与肿瘤坏死因子-α相结合的拮抗剂。首先,采用药效团模型和分子对接方法虚拟筛选specs商业库,然后,将得到的目标化合物进行表面等离子共振分析,得到4个不同骨架的肿瘤坏死因子-α结合配体。其中,T1与肿瘤坏死因子-α的亲和力最强(Kd=11μM),该亲和值与已有的肿瘤坏死因子-α小分子拮抗剂活性相当。进一步的细胞实验结果也表明T1与已知拮抗剂C87的活性相近。本研究不仅提供了一些肿瘤坏死因子-α小分子拮抗剂的新骨架,用于进一步的结构优化,同时也证明了虚拟筛选策略在肿瘤坏死因子-α小分子拮抗剂鉴别方面的实用性。2.基于代谢组学和网络药理学相结合的四逆汤抗阿霉素诱导心衰作用研究四逆汤是一味经典的治疗阿霉素诱导心衰的中药复方,由附子、干姜和甘草组成。目前的化学物质组学和药理学研究表明四逆汤主要的活性部位为附子总生物碱、干姜总姜酚、甘草总黄酮以及甘草总皂苷。本研究的动物实验结果表明四逆汤有效部位配伍治疗效果与四逆汤相近,比两两配伍或单组分配伍的疗效好。尽管四逆汤有效部位配伍的疗效明确,但是由于其成分的复杂性,其作用机制仍不明确。本研究旨在首次联用代谢组学和网络药理学方法探讨四逆汤有效部位配伍治疗心衰的潜在作用机制。通过基于GC/LC-MS的整体代谢组学策略,我们发现四逆汤有效部位配伍主要通过调控6个代谢通路发挥疗效。然后,我们进行了网络药理学研究,通过分子对接技术找到了有效部位作用的17个心血管疾病相关的潜在靶点,其中11个被文献证明为有效部位中化学成分的靶点,该结果也从网络药理学的角度证明了四逆汤有效部位整体的治疗效果。在找到的17个潜在靶标中,磷酸肌醇3-激酶γ、胰岛素受体、鸟氨酸转氨酶和葡糖激酶存在于四逆汤有效部位所调控的代谢通路中。并且,磷酸肌醇3-激酶γ、胰岛素受体和葡糖激酶已经被文献证明为四逆汤有效部位中化学成分的靶标。我们的研究也表明附子总生物碱是主要的活性成分,干姜总姜酚和甘草总黄酮总皂苷是辅助成分。代谢组学和网络药理学的联用为更全面可靠地挖掘中药化学物质组所作用的靶标群提供了可能,并为传统中药复方治疗作用的探讨提供了新的思路。3.基于网络分析的四逆汤活性成分的靶标鉴别及机制研究同步鉴别化学物质组的靶标群是当前面临的一个重要的且尚未解决的难题。在本研究中,我们创新性地整合了网络药理学和代谢组学数据,首次提出了网络分析的方法,用于同步鉴别四逆汤中抗心衰活性物质组的潜在靶标群。首先,我们通过血清药物化学、文本挖掘和相似性匹配相结合的方法鉴别出四逆汤抗心衰的48个潜在活性成分。然后采用文本挖掘和分子对接的方法鉴别这些活性成分的潜在靶标,并通过STRING数据库对这些靶标蛋白相关的生物过程,通路和相关疾病进行了富集分析。最后,整合代谢组学数据进行网络分析进一步筛选四逆汤活性成分的靶标蛋白,以提高文本挖掘和分子对接方法的准确性。在网络分析鉴别出的25个潜在靶标蛋白中,肿瘤坏死因子-α被首次验证为四逆汤中次乌头碱、新乌头碱、去甲乌药碱和槲皮素的靶标。表面等离子共振实验表明这些化合物能与肿瘤坏死因子-α直接结合。细胞实验结果也表明这些化合物能缓解肿瘤坏死因子-α导致的L929细胞毒性,并抑制阿霉素诱导的H9C2心肌细胞凋亡。我们期望网络分析也能被用于活性单体靶标的鉴别。4.HF-Net Pharm:心衰相关中草药网络药理平台的构建几千年来,作为多靶标疗法的代表,中药复方已经被成功地用于治疗多种疾病。然而,研究方法的缺乏极大地限制了其作用机制的探讨。本章首次构建了一个心衰相关中草药网络药理平台(HF-Net Pharm,www.cbligand.org/HF),用于中药复方作用机制的研究。该平台搜集了46个治疗心衰的中药复方、复方中111种药材以及药材中13,301个化合物的信息。同时,还整合了所有文献报道的心衰相关化学基因组学数据,包括259个心衰相关的靶标蛋白和77个治疗心衰的上市或临床试验药。为了对化合物的靶标进行预测,我们嵌入了Target Hunter和HTDocking模块。并且还对心衰相关网络药理平台的应用进行了举例,包括活性化合物的靶标鉴别,FDA批准抗心衰药物和草药抗心衰活性成分的多向药理分析,以及抗心衰中药复方的网络药理研究。通过文本挖掘和体外实验对预测结果进行了验证。本研究构建的心衰相关中草药网络药理平台将有助于中药材来源的药物发现、心衰相关的靶标鉴别、多向药理研究及中药复方的网络药理学研究。
[Abstract]:Heart failure is a common, expensive, and potentially fatal disease. In recent years, the prevalence and incidence of heart failure are increasing year by year. Heart failure disease has developed into a common public problem. This paper uses chemical informatics to virtual screening of small molecular antagonists targeting the target protein TNF - alpha targeting target protein, tumor necrosis factor - alpha. At the same time, the chemical informatics tool and the concept of network pharmacology are integrated to explore the potential mechanism of the anti heart failure of the four reverse soup, and a network pharmacology platform of the Chinese herbal medicine for heart failure is constructed, which provides a proprietary for the target identification of anti heart failure, the target identification of medicinal materials and compounds and the study of the network pharmacological mechanism. The network platform.1. predicts and verifies that TNF - alpha is a potential target protein for the treatment of heart failure based on the virtual screening of tumor necrosis factor - alpha small molecule antagonists. However, the number of small molecular inhibitors of TNF - alpha is very limited. Screening the antagonists with tumor necrosis factor alpha (TNF - alpha). First, the pharmacophore model and molecular docking method were used to selectively screen the specs commercial bank. Then, the target compounds were analyzed by surface plasmon resonance, and 4 tumor necrosis factor alpha binding ligands were obtained with different skeletons. Among them, T1 and tumor necrosis factor - alpha The affinity is the strongest (Kd=11 mu M), which is equivalent to the existing tumor necrosis factor - alpha small molecule antagonist. Further cell test results also show that the activity of T1 is similar to that of the known antagonist C87. This study not only provided some new skeleton of tumor necrosis factor - alpha small molecule antagonist, but also used for further structural optimization. The practicability of the virtual screening strategy in the identification of TNF - alpha small molecule antagonists.2. based on the combination of metabonomics and network pharmacology four inverse soup against adriamycin induced heart failure four reverse decoction is a classic Chinese herbal compound for the treatment of adriamycin induced heart failure, consisting of Aconitum, dried ginger and Glycyrrhiza. The previous study of chemical composition and pharmacology showed that the main active sites of the four reverse soup were total alkaloids of aconite, total ginger, total flavonoids and total liquorice saponins. The results of this study showed that the effective part of the four reverse soup was similar to the four reverse soup, and the effect was better than the compatibility of the 22 or the single component. The efficacy of the effective parts of the four reverse soup is clear, but its mechanism is still not clear due to the complexity of its components. The purpose of this study is to explore the potential mechanism of the effective part of the four reverse soup in the treatment of heart failure for the first time by means of metabonomics and network pharmacology. Through the overall metabolic strategy based on GC/ LC-MS, we found four The efficacy of the effective parts of the decoction is mainly controlled by the regulation of the 6 metabolic pathways. Then, we have conducted a network pharmacology study to find the potential targets of 17 cardiovascular diseases related to the effective parts of the site by molecular docking technique, and 11 of them were proved to be the targets of the chemical components in the effective part, and the results were also from the network. Pharmacologic points of view demonstrate the overall therapeutic effect of the effective parts of the four reverse soup. Among the 17 potential targets found, the inositol 3- kinase gamma, the insulin receptor, ornithine aminotransferase and glucokinase exist in the metabolic pathways regulated by the effective parts of the four reverse soup. And, the inositol phosphoric acid 3- kinase gamma, insulin receptor and glucokinase have already been found. Our study also showed that the total alkaloid of aconite is the main active ingredient, the total saponins of total ginger and total flavonoids of dried ginger are auxiliary components. The combination of metabolomics and network pharmacology is a more comprehensive and reliable target group for digging the chemical substance group of Chinese medicine more fully and reliably. It provides the possibility, and provides a new idea for the discussion of the therapeutic effect of traditional Chinese medicine compound. The target identification and mechanism of the active ingredient of the four inverse soup based on network analysis is an important and unsolved problem that the target group is facing at present. In this study, we have innovatively integrated it in this study. Network pharmacology and metabonomics data for the first time proposed a network analysis method for the simultaneous identification of the potential target groups of anti heart failure active substances in the four reverse soup. First, we identified the 48 potential active components of the anti heart failure of the four reverse soup through the combination of serum drug chemistry, text mining and similarity matching. Text mining and molecular docking methods identify the potential targets of these active components, and enrich the biological processes, pathways and related diseases related to these target proteins through the STRING database. Finally, the integrated metabolic data is integrated to screen the target proteins of the active ingredients of the four reverse soup to improve the text. The accuracy of this method of mining and molecular docking. In the 25 potential target proteins identified by network analysis, TNF - alpha was first verified as the target of aconitine, neoaconitine, noracinine and quercetin in four reverse soup. Surface plasmon resonance experiments showed that these compounds could be directly combined with tumor necrosis factor - alpha. The results of cell experiments also indicate that these compounds can alleviate the toxicity of tumor necrosis factor - alpha induced L929 cells and inhibit the apoptosis of H9C2 induced cardiomyocytes induced by adriamycin. We expect that network analysis can also be used for the identification of.4.HF-Net Pharm: heart failure related Chinese herbal medicine platform for thousands of years. The traditional Chinese medicine compound has been successfully used in the treatment of various diseases. However, the lack of research methods greatly restricts the discussion of its mechanism of action. In this chapter, a HF-Net Pharm (www.cbligand.org /HF) was first constructed for the study of the mechanism of the action of Chinese herbal compound. 46 compound Chinese medicine for the treatment of heart failure, 111 kinds of medicinal materials in the compound, and information of 13301 compounds in the medicinal materials were collected. At the same time, all the reports of heart failure related chemical genomics data, including 259 heart failure related target proteins and 77 listed or clinical trial drugs for the treatment of heart failure, were integrated. We embed the Target Hunter and HTDocking modules. We also exemplify the application of the pharmacological platform of the heart failure related network, including the target identification of active compounds, the multidirectional pharmacological analysis of the anti heart failure drugs and the anti heart failure active components of the herbal medicine by FDA, and the network pharmacology of the Chinese herbal compound of anti heart failure. The prediction results were verified by text mining and in vitro experiments. The network pharmacology platform of heart failure related herbal medicine constructed in this study will be helpful to the drug discovery of Chinese medicinal materials, the target identification of heart failure related, multi direction pharmacological research and the network pharmacology research of Chinese medicine compound.
【学位授予单位】：第二军医大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R285

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