标题: 分子间药理作用介面家族应用在磷酸化酵素-药物-疾病网络与机制之研究
Molecular Pharma-interface Families for Kinase-drug-disease Network and Mechanisms
作者: 杨进木
Yang, Jinn-Moon
国立交通大学生物科技研究所
关键字: 分子间药理作用介面家族;旧药新用;多标的药物;磷酸化酵素;癌症治疗;副作用;Molecular pharma-interface family;New uses for old drugs;Multi-target drugs;Kinase;Cancer treatment;Side effect
公开日期: 2014
摘要: 本五年期计划的主要目标在于利用所提出之新概念“分子间药理作用介面家族(molecular pharma-interface family)”结合“区域官能基地图(site-moiety map, SiMMap)”,以探讨蛋白磷酸化酵素抑制剂之结合机制与选择性。希望达到三项目标: (一)为上市药物与中草药寻找新用途(旧药新用)。(二)发展多标靶药物 (multi-target drugs)。(三)建立国人常见癌症之磷酸化酵素-药物-疾病关联性网络资料库,相信能藉此达到加速药物发展及增进治疗复杂疾病之效果。在上一期(第三年)中共发表了5篇科技论文 (Nucleic Acids Research,影响系数: 8.0;PLoS Comput Biol,影响系数: 4.9;BMC Bioinformatics,影响系数: 3.0;Basic & Clinical Pharmacology & Toxicology,影响系数: 2.1;BMC Genomics,影响系数: 4.4),并有2个美国专利申请中。
上一期中我们完成了以下事项:(一)蛋白磷酸化酵素-抑制剂-疾病家族地图资料库之建构。除可据此解释蛋白磷酸化酵素抑制剂之选择性与结合机制外,更可对对癌症等复杂疾病开发更有效的新型抑制剂(多标靶抑制剂)、克服耐药性与旧药新用。(二)以广泛出现于蔬果之类黄酮为核心,在此资料库的基础下预测不同方向延伸之类黄酮对蛋白磷酸化酵素之选择性,最后透过大规模之蛋白质磷酸化酵素-化合物之活性分析以验证此资料库。结果显示大部分的类黄酮均如此资料库的预测,只有JMY66在选择性上有偏差,须进一步检讨修正。同时,也确认了包含扁柏双黄酮 (JMY26)、槲皮素 (JMY27)、儿茶素 (JMY29)、黄芩素 (JMY51)、金鸡菊查尔酮苷 (JMY91)在内的多种类黄酮为蛋白磷酸化酵素抑制剂,其选择性从22%至56%不等,具潜力发展为癌症特效之多标靶抑制剂。并可由KIDFamMap预测其作用位向、结合机制与其选择性。另外透过同源映射网络推测JMY26有可能透过抑制IGF1R、KIT、RAF1与LCK等而影响细胞凋亡及透过抑制ABL1与LIMK1而影响轴突导向。(三)发现一个新颖的选择性蛋白磷酸化酵素抑制剂作用袋点,如类黄酮ZINC12872097与迷迭香酸,均因作用于此袋点而深具选择性,选择性分别为6%与3%。且作用于该袋点之化合物(如:迷迭香酸)有机会克服传统耐药性突变的影响,我们特将这类的化合物命名为C型 (type C) 选择性及抗耐药性抑制剂。为了更进一步验证此类抑制剂在临床治疗上的效果,已于八月时与淡水马偕医院合作申请【迷迭香酸治疗肺癌病人与其病理机制探讨】的临床试验,目前仍在审核中。(四)由单一的结构系统生物实验室茁壮成涵盖化学合成与新药物传输系统设计、生物化学、动物实验与临床医学等领域的成熟药物设计与系统生物团队,并积极与产业界合作:在与食工所的合作上,发现红麴中含有化合物JMY103能抑制多种蛋白磷酸化酵素,且对EGFR野生型及抗药型癌细胞皆具抑制能力,此初步结果将提升原有红麴商品之新商机;在与杰升生技的合作上,发现丹蔘含有化合物JMY102能有效抑制非小细胞肺癌的治疗标靶EGFR与类风湿性关节炎的潜在治疗标靶SYK,可望朝植物药新药努力;在与华联生技的合作上,由细胞与动物实验初步验证JMY-P01为新的乳癌生物标记,结合ERBB2等已知生物标记可有效侦测乳癌细胞之生成及类型,将可结合华联生技研发之晶片制造技术,开发癌症快筛检验试剂套组。
上述研究成果已达到计画去年之预期目标,初步成果显示相似化合物因具有相似交互作用与锚点而有类似的抑制效果的概念是可行的,并证实类黄酮可作为抑制剂之核心及藉由不同方向的延伸成为蛋白磷酸化酵素活化与否的开关。本年度的工作构想,主要分为三大部分:(一)将天然物由迷迭香酸与类黄酮扩展至药用植物,建立草药-天然物-作用蛋白质-疾病(症状) 网路,使之成为中草药药理平台。(二)将蛋白磷酸化酵素-抑制剂-疾病家族地图资料库与同源映射网路结合,发展以疾病为主的多标靶蛋白磷酸化酵素抑制剂,并与华联生技的强项-基因晶片结合,探讨蛋白磷酸化酵素抑制剂与细胞行为的关联,并从中寻找可作为疾病检验的潜在生物标记。(三)以广效型蛋白磷酸化酵素抑制剂为基础,藉由触及下游蛋白受质辨识锚点而创造选择性抑制剂(type S inhibitors),成为继以类黄酮为核心之后的另一类蛋白磷酸化酵素活化与否的开关。
The addressed issue of this project is to explore the binding mechanisms and kinase inhibitor selectivity by the novel concept, molecular pharma-interface family, and site-moiety map (SiMMap). There are three goal of this project including (1) new usages or targets of approved drugs and the Chinese herbs; (2) developments of multi-target drugs; (3) for popular diseases (such as cancers), to build the kinase-drug-disease network. It would be helpful to increase the drug developments and therapeutic effects of complex diseases. In previous period (third year), we have published 5 scientific papers (Nucleic Acids Research, impact factor: 8.0; PLoS Comput Biol, impact factor: 4.9; BMC Bioinformatics, impact factor: 3.0; Basic & Clinical Pharmacology & Toxicology, impact factor: 2.1; BMC Genomics, impact factor: 4.4) and two applied U.S.A patents.
During the period, we first constructed kinase-inhibitor-disease family map database (KIDFamMap). Though KIDFamMap database, we could elucidate the binding mechanisms and selectivity of kinase inhibitors. It also provides the opportunity to develop more effect inhibitors (multi-target drugs) for complex diseases (such as cancers), anti-drug resistance, new uses of existing drugs. Then, we focused on the flavonoid derivatives, which are a class of plant secondary metabolites and have over 8000 derivatives. These derivatives have the same core structure and different extended functional groups. Based on KIDFamMap and kinase-inhibitor family, the interactions and selectivity of flavonoid derivatives were predicted. The large-scale kinase-inhibitor profiling was used to verify the results. The experiments confirmed the predictions, excluding JMY66. The flavonoid derivatives, including hinokiflavone (JMY26), quercetin (JMY27), catechin (JMY29), baicalein (JMY51), marein (JMY91) were confirmed as kinase inhibitors (selectivity from 22% to 56%) and KIDFamMap also provided the binding models and mechanisms of these derivatives. It could be potential inhibitor for multiple targets for cancers. JMY26 may induce cancer cell apoptosis by inhibiting kinase IGF1R, KIT, RAF1 and LCK and affect axon guidance by inhibiting kinase ABL1 and LIMK1 through homologous mapping networks predictions. Moreover, we identified a novel selective binding pocket for kinase-inhibitor interactions and some inhibitors bind this pocket, including flavonoid derivative ZINC12872097 and rosmarinic acid. The kinase selectivity are 6% and 3% for ZINC12872097 and rosmarinic acid, respectively. The inhibitors bind the selective pocket can overcome the effects of drug resistant mutations and are called as Type C inhibitors. For testing the therapeutic effect of clinical trials of these Type C inhibitors, we cooperated with Mackay Memorial Hospital and applied the Institutional Review Board (IRB) of “Treatment of lung cancer with rosmarinic acid”. In addition, we cooperated with other laboratories to form a drug design and systems biology research team involved chemical synthesis, drug delivery, biochemistry, animal experiments, and clinical trials. We cooperated with The Food Industry Research and Development Institute (FIRDI) and found the compound (JMY103) in Monascus purpureus can target many protein kinases and inhibit both EGFR wild-type and EGFR with drug-resistant mutations. With Jason Life Tech Inc., compound (JMY102) of Salvia miltiorrhiza was found to target EGFR related to non-small-cell lung carcinoma (NSCLC) and SYK related to rheumatoid arthritis and can be potential herbal medicine. Cooperated with Phalanx Biotech, we have identified JMY-P01 as new biomarker of breast cancer and verified by cell and animal experiments. By combining known biomarkers, such as ERBB2, and it can be utilized to develop cancer diagnostic kits with Phalanx Biotech to detect breast cancer.
Our results show that similar compounds with similar interactions will perform similar inhibitions and use flavonoid derivatives to explore the binding mechanisms and selectivity. The goals of this period are described as following: (1) we will focus on herbs and create herb-natural product-target-disease network and herb pharmacological platform. (2) Combining kinase-inhibitor-disease family map database and homologous mapping network to develop disease-driven kinase inhibitors and study the relationships between kinase inhibitors and cell-behaviors though gene chips by cooperating with Phalanx Biotech. (3) We propose a new kind of selective inhibitors (called Type S inhibitors) by extending broad-spectrum the inhibitors to kinase substrate binding pocket. We will develop highly selective Type S inhibitors and study kinase-inhibitor binding mechanisms.
官方说明文件#: NHRI-EX103-10009PI
URI: http://hdl.handle.net/11536/101801
https://www.grb.gov.tw/search/planDetail?id=8049082&docId=427688
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