清热解毒解表类中药辅助治疗流感的分子机制研究

The prevention and the treatment of influenza are the major issues for human health. The clinical drug therapy is mainly western medicine supplemented by Traditional Chinese Medicine (TCMs) at the present stage. In the clinics, influenza is mainly treated with western medicine of neuraminidase (NA) inhibitor drugs. However, the emergence of drug-resistant NA variants is the major problems of antiviral therapy. TCMs treatment of influenza has a long history, its resolves both the symptom and cause of the disease with minimal side effects, effectively adjust and balance the state of the human body. But the treatment integrated Chinese and Western medicine lack of in-depth study on the mechanism at present. Thus research and discovery of new anti-NA strategy is of particularly importance. In this project, we will screen compounds from exterior-releasing, heat-clearing, and detoxicating medicinal of TCMs library using computer simulation and enzyme kinetics method. Study on the compounds which have NA inhibitory activities and reversal NA resistance combined with western medicine. Molecular docking and molecular dynamics method will be used to study the type of the structure-activity relationships of NA and the active ingredient to reveal the mechanism of action. Furthermore, test the TCMs and Chinese Medicine containing the NA inhibitory compounds with similar efficacy. Finally, the anti-influenza virus effect of the active ingredient will be evaluated in vitro and in vivo. The ultimate goal of this project is to reveal the molecular mechanism of TCMs as an adjunctive therapy with western medicine in anti-influenza and reversal drug resistance. It will contribute to help drug selection of combined traditional Chinese medicine and western medicine in clinics, and to provide the new effective clinical solutions for influenza treatment.

流感对人类健康造成重大危害，现阶段的临床用药方案主要是以西药为主中药为辅的方式。西药以神经氨酸酶(NA)抑制剂类药物为主，但因NA变异而造成的耐药性问题，对部分患者疗效不佳。中药抗流感历史悠久，具有标本兼治、调理人体整体状态的独特优势。而中西医结合治疗流感疗效更加显著，但其背后的机制亟待阐明。我们发现，一些中药单体可抑制NA并且增强西药的疗效，较好地解释了中西医结合对流感的治疗作用。本项目拟以计算机模拟和酶学实验检测方法相结合，从清热解毒解表类中药单体文库中，筛选得到与西药协同作用提高NA抑制活性并逆转突变型NA耐药性的中药单体。通过分子对接等方法，分析联合药物抑制NA的作用机制。在此基础上，检测含有与西药协同作用单体的中药以及中药复方，并对其抗流感病毒效果进行细胞和动物水平的评价。本项目的实施，有助于解释中西医结合治疗流感并逆转病毒耐药性的分子机制，并为临床选择用药提供新的思路和方法。

流行性感冒是由流感病毒引起的一种具有高度传染性的急性呼吸道疾病。对人类健康和生命造成巨大危害，同时也给全球的经济带来巨大的损失，是人类至今尚不能控制的世界性传染病之一。抗流感药物一直是治疗流感的重要有效途径，目前应该比较广泛的流感病毒治疗靶点有神经氨酸酶和核酸内切酶。但是，突变体出现使得流感病毒对药物产生耐药性，这对我们研发新型的抗流感病毒药物以及解决流感病毒的耐药性成为研究抗流感病毒药物的重点和热点。中医抗流感历史悠久，积累的大量行之有效的抗流感中药和中药复方，中药具有不同于西药的抗流感方式，中药和中药复方往往能够作用于流感病毒复制过程中的多个靶点，并且很多中药能够通过调节机体免疫力来治疗流感。这使得运用中药解决流感病毒的耐药性成为可能，但是对于中药成分抗流感病毒作用的机制还缺乏系统深入的研究。我们通过分子模拟和酶学实验检测方法相结合，从中药单体数据库中，筛选得到有效的流感病毒抑制剂。获得含有中药单体的中药，对其进行实验上的抑制酶活效果测定，以鉴定其酶学抑制效应。随着本项目的实施，我们筛选得到了中药单体芦丁、金丝桃苷、水仙苷、杨梅苷、洋蓟素和1,5-二咖啡酰奎宁酸对神经氨酸酶有较好的抑制效果；发现淫羊藿和韭菜籽对无论是流感病毒核算内切核酸酶的野生型还是PA-I38T突变型都有很好的水解活性；以及应用老药新用的思路得到钙黄绿素具有很好的抑制神经氨酸酶效果，并且可以和奥司他韦联合用药具有协同作用。以上项目的实施，有助于解释中西医结合治疗流感并逆转病毒耐药性的分子机制，并为临床选择用药提供新的思路和方法。