GAPDH抑制剂的设计与合成及功能研究

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a multi-functional protein in the cell. It has attracted much attention in recent years as a potential drug target. Our preliminary study for the first time found that GAPDH binded with cADPR, and mediated cellular calcium signaling pathway which was induced by cADPR. In this research, we will develop specific inhibitor which targets to GAPDH/cADPR binding site with the aid of computer aided drug design and other strategies. Meanwhile, in vitro and in vivo activities will be studied, molecular mechanism of action will be investigated, and the drugability will be evaluated. This research will expand the understanding of the biological function of GAPDH, have a deeper understanding for possibility of GAPDH as drug targets, and promote the development of GAPDH targeted antitumor drug candidates and drug target validation. The details of this research mainly includes: 1) the discovery of active compounds based on virtual screening, compound library assay, and structural modification of known compound. 2) in vitro screening based on calcium signaling, SPR and inhibition of proliferation of tumor cells. 3) development of synthetic strategy, structural optimization, and summarization of structure-activity relationship. 4) investigation of molecular mechanism from the aspects of effects on calcium pathway, PI3K/Akt/mTOR pathway, and Oxidative stress reaction. 5) the evaluation of drugability based on pharmacodynamics, pharmacokinetics and safety studies.

GAPDH是细胞内功能众多的一种蛋白，近年来作为药物潜在靶标备受关注。本课题组首次发现了GAPDH与cADPR结合、并介导cADPR调控的细胞钙信号通路，本项目拟借助计算机辅助药物设计等策略，发展靶向GAPDH/cADPR结合位点的GAPDH抑制剂，并进行体内外活性、作用机制及成药性评价。本研究将拓展对GAPDH的生物学功能的认识，促进对GAPDH作为药物靶标的可能性的了解，为发展抗肿瘤药物以及药物靶标的确认奠定基础。研究内容主要包括：1基于虚拟筛选和实体化合物库筛选以及已知结构改造的活性化合物的发现；建立在细胞钙信号影响、表面等离子共振信号变化及肿瘤细胞增殖抑制的体外活性评价；发展合成方法、结构优化及构效关系研究；探讨抑制剂对细胞钙信号通路、PI3K/Akt/mTOR通路、氧化应激反应及肿瘤细胞侵袭迁移的影响；5) 从药效、药代及安全性方面初步考察抑制剂及靶点的成药性

本项目围绕cADPR参与调控钙信号通路，项目聚焦于小分子调节剂的设计、合成、机制与功能研究，完成以下主要研究内容：a) 深入研究了GAPDH对cADPR钙信号通路的调控机制，考察了作用位点，研究证实了GAPDH是cADPR的特异结合蛋白，并且cADPR与GAPDH的结合是一种瞬时结合。b) 对cADPR通过TRPM2调控钙信号通路的分子机制进行了深入研究，借助分子模拟方法并辅以点突变与SPR结合实验，解析了关键作用位点；借助计算机辅助药物设计策略，设计、合成并获得了对TRPM2具有特异抑制作用的化合物，并经初步成药性评价获得了高活性药物先导化合物。c) 发展了基于神经网络的深度学习分子图生成方法，所设计的药物分子具有结构多样性与可合成性及成药性特征；发展了基于分子三维力场的3D分子指纹TF3P策略的深度神经网络药物分子多靶标预测方法。d) 基于cADPR与GAPDH的结合模式分析，通过虚拟筛选获得了多个GAPDH功能调控分子，考察了个别分子对cADPR钙信号的调控作用，并通过结构优化活性得到了提升。在本项目执行期间，共发表研究论文11篇，获中国发明专利授权1项，应邀做会议学术报告7次；博士研究生毕业4人、硕士研究生毕业5人、博士后出站2人。项目整体完成了计划目标。