TITLE OF PROJECT : Invention of intensifiers of antibiotics-susceptibility against multi-drugs resistant pathogenic bacteria and invesrigation for novel control system of multi-drugs resistance

项目名称：多重耐药病原菌抗生素敏感性增强剂的发明及新型多重耐药控制体系的研究

• 批准号: 14390038
• 负责人: HIGUTI Tomihiko
• 金额: $ 8.51万
• 依托单位: The University of Tokushima
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14390038/
• 关键词: medicinal plants; MRSA; intensifier of β-lactam-susceptibility; high throughput screening; alkyl gallate; structure-activity relationship; flavone; two-components signal transduction system; 生薬; 岩白菜; 最適炭素鎖長; 親水性基と疎水性基; 生薬植物; β-ラクタム剤-感受性増強薬

High-throughput screening for novel intensifiers of β-lactam-susceptibility in MRSAs (ILSMRs)We were screening for the activity of 900 species of medicinal plants to modulate β-lactam resistance of methicillin-resistant Staphylococcus aureus (MRSA). We found 43 species of plants which showed the potent antibacaterial activity and the same number of plants which had the ILSMR activity. Among them, a methanol extract of fruit pods of Caesalpinia spinosa (Tara) showed a potent ILSMR activity. By the activity-guided fractionation of the extracts, we purified and identified four compounds including ethyl gallate as effective constituents. SAR analysis with various alkyl gallates and the related compounds revealed the significant contribution of the galloyl moiety and the alkyl chain. The optimum alkyl chain lengths were shown at C9-10 for the antibacterial activity and at C5-6 for the ILSMR activityWe also found that Bergenia purpurascens (Gan-hakusai) showed remarkable activity. The isolation and identification procedure of active principles from the fraction is in progressStudy on the mechanism of action of novel ILSMRsWe detected 16 up-regulated clones, including the genes associated with fatty acid metabolism and those encoding the two-component signal transduction system (TCSTS), and 10 down-regulated clones, including the genes associated with metabolic pathways such as the tricarboxylic acid cycle and oxidative phosphorylation. A positive correlation was found between the up-regulation of the TCSTS and the β-lactam susceptibility intensified by flavone in the MRSA strains, and disruption of the gene encoding the sensor of the TCSTS resulted in decreased oxacillin resistance. The results suggest important roles of the TCSTS in resistance to β-lactam antibiotics

高通量筛选新型 MRSA β-内酰胺敏感性增强剂 (ILSMR) 我们正在筛选 900 种药用植物调节耐甲氧西林金黄色葡萄球菌 (MRSA) β-内酰胺耐药性的活性。我们发现了 43 种植物表现出有效的抗菌活性，并且有相同数量的植物具有 ILSMR 活性。其中，Caesalpinia spinosa (Tara) 果荚的甲醇提取物显示出有效的 ILSMR 活性。通过提取物的活性引导分馏，我们纯化并鉴定了包括没食子酸乙酯在内的四种化合物作为有效成分。对各种没食子酸烷基酯和相关化合物的 SAR 分析揭示了没食子酰基部分和烷基链的显着贡献。抗菌活性的最佳烷基链长度为 C9-10，ILSMR 活性的最佳烷基链长度为 C5-6。我们还发现紫白白菜 (Gan-hakusai) 显示出显着的活性。该馏分中有效成分的分离鉴定工作正在进行中。新型ILSMR的作用机制研究，检测到16个上调克隆，包括与脂肪酸代谢相关的基因和编码双组分信号转导系统（TCSTS）的基因，以及10个下调克隆，包括与三羧酸循环和氧化等代谢途径相关的基因。 磷酸化。研究发现，MRSA 菌株中 TCSTS 的上调与黄酮增强的 β-内酰胺敏感性之间呈正相关，而 TCSTS 传感器编码基因的破坏导致苯唑西林耐药性降低。结果表明 TCSTS 在 β-内酰胺抗生素耐药性中发挥重要作用

项目成果

姫田 敏樹: "「新ミトコンドリア学」.ミトコンドリアのバイオジェネシス"共立出版. 4 (2001)

Toshiki Himeda：“‘新线粒体学’。线粒体生物发生”Kyoritsu Shuppan 4 (2001)。

Kawazoe, K., et al.: "Sesquiterpenoids from Artemisia gilvescens and an anti-MRSA compound."Journal of Natural Products. 66(4). 538-539 (2003)

Kawazoe, K. 等人：“来自银蒿的倍半萜类化合物和抗 MRSA 化合物。”天然产物杂志。

Shibata A.: "Photocurrent of Purple Membrane Adsorbed onto a Thin Polymer Film : Effects of Monovalent and Divalent Ions"Studies in Surface Science and Catalysis. 132. 635-637 (2001)

Shibata A.：“吸附在聚合物薄膜上的紫色膜的光电流：单价和二价离子的影响”表面科学和催化研究。

Kondo, K., et al.: "ILSMRs (Intensifier of β-Lactam-Susceptibility in Methicillin-Resistant Staphylococcus aureus) from Tara [Caesalpinia spinosa (Molina) Kuntze]"Phytomedicine. 11(in press). (2004)

Kondo, K. 等人：“ILSMR（耐甲氧西林金黄色葡萄球菌的 β-内酰胺敏感性增强剂），来自 Tara [Caesalpinia spinosa (Molina) Kuntze]”植物医学 11（印刷中）。

堀尾武一監修: "分子細胞生物学基礎実験法 改定第2版"南江堂. 37 (2004)

堀尾武一监督：“分子和细胞生物学的基本实验方法，修订第2版”Nankodo 37（2004）。