Structure analyses of key enzymes from trypanosoma and malaria for the development of new drugs

用于新药开发的锥虫和疟疾关键酶的结构分析

• 批准号: 19370043
• 负责人: YOSHIMOTO Tadashi
• 金额: $ 11.56万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2007
• 资助国家: 日本
• 起止时间: 2007 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-19370043/
• 关键词: マラリア; トリパノソーマ; オリゴペプチダーゼB; 阻害剤; 医薬品; アミノペプチダーゼ; メチオニンアミノペプチダーゼ; 治療薬; 構造生物学; 酵素; 立体構造; X線結晶構造解析; プロテアーゼ; 感染症; 結晶解析

Since there is no effective medicine for infectious disease caused by Trypanosoma and malaria, many peoples live in tropical were killed. In order to develop new drugs for these, we focused to metabolic enzymes in Trypanosoma and malalia as target, and studied specific inhibitors.(1) Methionine aminopeptidase from P. falciparum ; We develop new assay method of methionine aminopeptidase using prolyl aminopeptidase. We cloned the enzyme gene and expressed in E. coli. Using the new assay method, specific inhibitor have been screened. A family of structurally related inhibitors containing a 2-(2pyridinyl)- pyrimidine was identified in compound library. Inhibitors were also found in microbial products and plant extract.(2) Oligopeptidase B from trypanosomiasis : The enzyme plays a key role in the pathogenesis of trypanosomiasis. We found protamine act unique and potent inhibitor for oligopeptidase B from trypanosoma. In order to clarify the mechanism of inhibition, we crystallized the enzyme and studied X-ray crystallography. The crystallographic parameters were a=b=123.9, C=248.0 A gamma=120 degree and p hexagonal.(3) Specific inhibitors for aminopeptidase N and prolyl oligopeptidase from them were also studied.

由于对锥虫病和疟疾引起的传染病没有有效的药物，许多生活在热带地区的人因此丧生。为了开发针对这些疾病的新药，我们重点以锥虫和疟疾的代谢酶为靶点，研究特异性抑制剂。(1)恶性疟原虫甲硫氨酸氨基肽酶；建立了以脯氨酸氨基肽酶为检测手段测定蛋氨酸氨基肽酶的新方法。我们克隆了该酶基因并在大肠杆菌中表达。利用新的测定方法，筛选了特异性抑制剂。在化合物文库中发现了一个含有2-（2吡啶基）-嘧啶的结构相关抑制剂家族。在微生物产物和植物提取物中也发现了抑制剂。(2)锥虫病寡肽酶B：该酶在锥虫病的发病机制中起关键作用。我们发现鱼精蛋白是一种独特而有效的锥虫寡肽酶B抑制剂。为了明确抑制机制，我们对酶进行了结晶和x射线晶体学研究。结晶参数为a=b=123.9， C=248.0, a=120°，p为六边形。(3)对氨基肽酶N和脯氨酰寡肽酶的特异性抑制剂也进行了研究。

项目成果

Structure of aminopeptidase N from Escherichia coli complexed with the transition state alnalogue aminophosphinic inhibitor PL250, Marie-Claude Fournie-Zaluski, Herve Poras and Bernard P.Roques

大肠杆菌氨肽酶 N 与过渡态类似物氨基膦抑制剂 PL250 复合的结构，Marie-Claude Fournie-Zaluski、Herve Poras 和 Bernard P.Roques

• 发表时间: 2009
• 期刊: Acta Crystal.D. 65
• 作者: 中嶋義隆;伊藤潔;芳本忠
• 通讯作者: 芳本忠

Closed Complex of the D-3-Hydroxybutyrate Dehydrogenase Induced by an Enantiomeric Competitive Inhibitor : Importance of Gln196 in Stable Ternary Complex Formation.

对映体竞争性抑制剂诱导的 D-3-羟基丁酸脱氢酶的闭合复合物：Gln196 在稳定三元复合物形成中的重要性。

• 发表时间: 2009
• 期刊: J.Biochem. 145,(4)
• 作者: Kanako Nakashima;Kiyoshi Ito;Yoshitaka Nakajima;Ryuji Yamazawa;Syunsuke. Miyakawa;Tadashi Yoshimoto
• 通讯作者: Tadashi Yoshimoto

微生物酵素の生化学的および構造生物学的研究と医療への応用

微生物酶的生化和结构生物学研究及医学应用

• 发表时间: 2007
• 期刊: 薬学雑誌 127
• 作者: Kasuya;E.;芳本 忠
• 通讯作者: 芳本 忠

Crystal Structure of FAD-dependent D-Glucose Dehydrogenase complexed with an inhibitor

FAD 依赖性 D-葡萄糖脱氢酶与抑制剂复合的晶体结构

• 发表时间: 2010
• 作者: 中嶋義隆;西矢芳昭;芳本忠;伊藤潔
• 通讯作者: 伊藤潔

Closed Complex of the D-3-Hydroxybutyrate Dehydrogenase Induced by an Enantiomeric Competitive Inhibitor

对映体竞争性抑制剂诱导的 D-3-羟基丁酸脱氢酶的闭合复合物

• 发表时间: 2009
• 期刊: J. Biochem. 145(4)
• 作者: Kanako Nakashima;Kiyoshi Ito;Yoshitaka Nakajima;Ryuji Yamazawa;Syunsuke Miyakawa;and Tadashi;Yoshimoto
• 通讯作者: Yoshimoto