现代中药创制设计方法学及关键技术研究
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摘要
我国拥有极丰富的传统中药资源,是研发现代中药的重要战略资源。目前中药新药研发通常采用三种研究模式:一种是以临床有效的经方、验方按传统工艺研制成中成药;第二种是通过分离分析与药效筛选,找出中药的有效部位或有效单体化合物制成中药新药;第三种是在已有成方制剂基础上进行精制开发。这些中药新药研制方法技术水平偏低,缺乏新药设计技术支持。因此,发展现代中药新药发现与设计技术是我国医药领域的重大战略需求和科学研究前沿。
如何从传统中药中发现有效组分,进而通过有效组分的配伍优化,设计出化学组成与作用机制相对明确的现代中药,是当前中药新药创制领域所面临的重要研究命题。本论文针对现代中药创制中的主要技术环节开展创新研究,将分析技术、生物技术与计算智能技术等相结合,创新发展了用于创制现代中药的多组分药物发现和优化设计方法,并成功应用于气血并治方、双丹方、复方丹参方等方剂的二次开发研究,发展形成了以方剂中有效组分创制现代中药的新模式。主要研究工作及创新研究成果包括:
1.提出一种中药有效组分智能辨识方法。该方法将中药分离分析、药效评价实验与因果关系发现技术相结合,通过对化学组成与药效评价数据进行相关性分析,计算辨析出中药药效物质。该方法应用于研究人参提取物,辨识出具有抑制肿瘤细胞增殖作用的人参皂苷类成分;应用于研究双丹方,辨识出具有心肌保护作用的活性化合物;还将其用于研究气血并治方,发现了具有降脂作用的有效组分。应用研究结果表明,该方法能利用化学组成与药效评价数据较为准确地从药材或中药复方中寻找到有效组分或成分,为从传统中药中发现药效物质提供了一种新的技术手段。
2.提出了基于定量组效关系建模的中药有效组分配伍优化设计方法。该方法通过建立中药组分配伍与药效间相关性定量模型,定量描述中药组分配伍与药效活性的相随变动关系,在此模型上用优化搜索算法对所有的组分配伍方案进行排序寻优,可设计得到有效组分的优化配伍。在气血并治方有效组分配伍优化中的应用结果表明,该方法可根据降脂及抗炎等指标设计得到药效优于原方的组分中药。
3.研究建立了中药蛋白质组分析方法。运用该方法研究了双丹方抗急性缺血心肌作用机制,发现给药前后大鼠缺血心肌的23个差异蛋白,包括与呼吸链、三羧酸循环和β氧化等能量代谢相关的11个蛋白以及超氧化物岐化酶、Hsp27等与细胞氧化应激相关蛋白。通过对差异蛋白的功能分析,揭示了双丹方通过改善能量代谢、对抗氧化应激等途径抗急性心肌缺血的作用机制。
4.运用中药蛋白质组分析方法研究复方丹参方保护心肌作用机制,鉴定出32个差异蛋白,并研究了复方丹参方对氧化应激相关的Hsp27蛋白表达及其翻译后修饰的影响。研究发现复方丹参方可通过调节能量代谢、抗氧化应激、保护线粒体功能和改善细胞结构等多个途径发挥心肌保护作用。
Traditional Chinese medicine (TCM) is a great treasure-house in China, which has become important resources for drug discovery and design. Nowadays, there are three major strategies for the development of Chinese medicines from TCM. The first strategy is to manufacture botanical drug from clinically therapeutical prescription by traditional preparation techniques. The second method is to isolate, screen and identify active components or chemical compounds from TCM, which can be used as herbal medicine. The third approach is to develop new drug from already marketed drugs by changing the formulation or clinical use. However, the technological levels of those methods are relatively low, especially lacks of appropriate methodology and key techniques for drug design. Therefore, the establishment of new technology for drug discovery and design has become pivotal industrial demands and scientific research project in Chinese pharmaceutical fields.
The major challenges in the discovery and design of modern drug from TCM includes three major problems, i.e. how to discover active components from TCM, how to design optimal combination of active components, and how to dissect the chemical composition and exact mechanism of action of those components. In this dissertation, a series of techniques were established aiming to solve those problems in the drug development pipeline of modern Chinese medicine. By integrating multi-disciplinary technologies from analytical chemistry, biochemistry and computer sciences, novel methodology and techniques for the design of modern Chinese medicine have been established. The proposed methods have been successfully applied in the re-discovery of many traditional prescriptions including Qi-Xue-Bing-Zhi-Fang, Shuangdan Decoction, and Compound Danshen, which indicated a new mode for the development of multi-component drug using active components of traditional formulae. The major results obtained from the study have been described as following:
1. A novel method was proposed to identify active components of TCM. This method used a stepwise causal adjacent relationship discovery algorithm to analyze correlation between the composition and bioactivity of TCM. Integrated with phytochemical analytical and bioassay techniques, causality discovery algorithm can find out active components from the complex mixture. Successful applications of this method have been performed in discovering anti-cancer compounds from mixture of ginsenosides, cardio-protective compounds from Shuangdan Decoction, and identifying components combination with decreasing total cholesterol activity from Qi-Xue-Bing-Zhi-Fang. The results indicated that the present method can accurately discover active components or compounds based on the composition and bioactivity data of TCM, which offers a new way to screen active components from TCM.
2. The strategy for designing multi-component drugs from TCM was developed by modeling quantitative composition-activity relationship (QCAR). The computational models characterized the relationship between chemical composition and their activities of components, which can be used to predict activity of all possible combination of components and to select the optimal combination of multiple active components by exhaustive searching and genetic algorithm. This approach was successfully applied in finding optimal combination of two active components of Qi-Xue-Bing-Zhi-Fang. Validation experiments showed that the optimized combination has greater activity than original remedy, which implies that the presented method is an efficient approach for the design of botanical drug from TCM.
3. The analytical approach for the pharmacoproteomics study of TCM was established. The proposed comparative proteomic approach was applied to investigate the potential mechanism of action for cardioprotective effects of Shuangdan Decoction (SDD) on ischemic rat heart. Twenty-three altered proteins were identified. Those proteins included eleven proteins related to energy metabolism process, such as respiration chain, TCA-cycle, andβ-oxidation. Moreover, some proteins linked to oxidative stress such as SOD and Hsp27 were also found to be regulated by SDD. Functional analysis of the differentially expressed proteins suggested that SDD diminished ischemic-induced damage through promoting myocardial energy status and reducing oxidative stress.
4. Cardioprotective effect of Compound Danshen on coronary artery ligated rats has been studied by the proposed proteomic approach. Thirty-two differentially expressed proteins were identified. Moreover, expression of heat shock protein 27 (Hsp27) and related post-translational modifications (PTMs) induced by Compound Danshen were further studied. The results suggested that the mechanism of action of Compound Danshen was related to the improvement of energy metabolism, recovery of mitochondrial function, and the depressing of oxidative stress induced injury in ischemia myocardium.
如何从传统中药中发现有效组分,进而通过有效组分的配伍优化,设计出化学组成与作用机制相对明确的现代中药,是当前中药新药创制领域所面临的重要研究命题。本论文针对现代中药创制中的主要技术环节开展创新研究,将分析技术、生物技术与计算智能技术等相结合,创新发展了用于创制现代中药的多组分药物发现和优化设计方法,并成功应用于气血并治方、双丹方、复方丹参方等方剂的二次开发研究,发展形成了以方剂中有效组分创制现代中药的新模式。主要研究工作及创新研究成果包括:
1.提出一种中药有效组分智能辨识方法。该方法将中药分离分析、药效评价实验与因果关系发现技术相结合,通过对化学组成与药效评价数据进行相关性分析,计算辨析出中药药效物质。该方法应用于研究人参提取物,辨识出具有抑制肿瘤细胞增殖作用的人参皂苷类成分;应用于研究双丹方,辨识出具有心肌保护作用的活性化合物;还将其用于研究气血并治方,发现了具有降脂作用的有效组分。应用研究结果表明,该方法能利用化学组成与药效评价数据较为准确地从药材或中药复方中寻找到有效组分或成分,为从传统中药中发现药效物质提供了一种新的技术手段。
2.提出了基于定量组效关系建模的中药有效组分配伍优化设计方法。该方法通过建立中药组分配伍与药效间相关性定量模型,定量描述中药组分配伍与药效活性的相随变动关系,在此模型上用优化搜索算法对所有的组分配伍方案进行排序寻优,可设计得到有效组分的优化配伍。在气血并治方有效组分配伍优化中的应用结果表明,该方法可根据降脂及抗炎等指标设计得到药效优于原方的组分中药。
3.研究建立了中药蛋白质组分析方法。运用该方法研究了双丹方抗急性缺血心肌作用机制,发现给药前后大鼠缺血心肌的23个差异蛋白,包括与呼吸链、三羧酸循环和β氧化等能量代谢相关的11个蛋白以及超氧化物岐化酶、Hsp27等与细胞氧化应激相关蛋白。通过对差异蛋白的功能分析,揭示了双丹方通过改善能量代谢、对抗氧化应激等途径抗急性心肌缺血的作用机制。
4.运用中药蛋白质组分析方法研究复方丹参方保护心肌作用机制,鉴定出32个差异蛋白,并研究了复方丹参方对氧化应激相关的Hsp27蛋白表达及其翻译后修饰的影响。研究发现复方丹参方可通过调节能量代谢、抗氧化应激、保护线粒体功能和改善细胞结构等多个途径发挥心肌保护作用。
Traditional Chinese medicine (TCM) is a great treasure-house in China, which has become important resources for drug discovery and design. Nowadays, there are three major strategies for the development of Chinese medicines from TCM. The first strategy is to manufacture botanical drug from clinically therapeutical prescription by traditional preparation techniques. The second method is to isolate, screen and identify active components or chemical compounds from TCM, which can be used as herbal medicine. The third approach is to develop new drug from already marketed drugs by changing the formulation or clinical use. However, the technological levels of those methods are relatively low, especially lacks of appropriate methodology and key techniques for drug design. Therefore, the establishment of new technology for drug discovery and design has become pivotal industrial demands and scientific research project in Chinese pharmaceutical fields.
The major challenges in the discovery and design of modern drug from TCM includes three major problems, i.e. how to discover active components from TCM, how to design optimal combination of active components, and how to dissect the chemical composition and exact mechanism of action of those components. In this dissertation, a series of techniques were established aiming to solve those problems in the drug development pipeline of modern Chinese medicine. By integrating multi-disciplinary technologies from analytical chemistry, biochemistry and computer sciences, novel methodology and techniques for the design of modern Chinese medicine have been established. The proposed methods have been successfully applied in the re-discovery of many traditional prescriptions including Qi-Xue-Bing-Zhi-Fang, Shuangdan Decoction, and Compound Danshen, which indicated a new mode for the development of multi-component drug using active components of traditional formulae. The major results obtained from the study have been described as following:
1. A novel method was proposed to identify active components of TCM. This method used a stepwise causal adjacent relationship discovery algorithm to analyze correlation between the composition and bioactivity of TCM. Integrated with phytochemical analytical and bioassay techniques, causality discovery algorithm can find out active components from the complex mixture. Successful applications of this method have been performed in discovering anti-cancer compounds from mixture of ginsenosides, cardio-protective compounds from Shuangdan Decoction, and identifying components combination with decreasing total cholesterol activity from Qi-Xue-Bing-Zhi-Fang. The results indicated that the present method can accurately discover active components or compounds based on the composition and bioactivity data of TCM, which offers a new way to screen active components from TCM.
2. The strategy for designing multi-component drugs from TCM was developed by modeling quantitative composition-activity relationship (QCAR). The computational models characterized the relationship between chemical composition and their activities of components, which can be used to predict activity of all possible combination of components and to select the optimal combination of multiple active components by exhaustive searching and genetic algorithm. This approach was successfully applied in finding optimal combination of two active components of Qi-Xue-Bing-Zhi-Fang. Validation experiments showed that the optimized combination has greater activity than original remedy, which implies that the presented method is an efficient approach for the design of botanical drug from TCM.
3. The analytical approach for the pharmacoproteomics study of TCM was established. The proposed comparative proteomic approach was applied to investigate the potential mechanism of action for cardioprotective effects of Shuangdan Decoction (SDD) on ischemic rat heart. Twenty-three altered proteins were identified. Those proteins included eleven proteins related to energy metabolism process, such as respiration chain, TCA-cycle, andβ-oxidation. Moreover, some proteins linked to oxidative stress such as SOD and Hsp27 were also found to be regulated by SDD. Functional analysis of the differentially expressed proteins suggested that SDD diminished ischemic-induced damage through promoting myocardial energy status and reducing oxidative stress.
4. Cardioprotective effect of Compound Danshen on coronary artery ligated rats has been studied by the proposed proteomic approach. Thirty-two differentially expressed proteins were identified. Moreover, expression of heat shock protein 27 (Hsp27) and related post-translational modifications (PTMs) induced by Compound Danshen were further studied. The results suggested that the mechanism of action of Compound Danshen was related to the improvement of energy metabolism, recovery of mitochondrial function, and the depressing of oxidative stress induced injury in ischemia myocardium.
引文
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