糖加水分解酵素蛍光プローブ群の開発による新規迅速がんイメージング手法の確立

开发一组糖基水解酶荧光探针，建立新的快速癌症成像方法

• 批准号: 16F16109
• 负责人: 浦野 泰照
• 金额: $ 1.41万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for JSPS Fellows
• 财政年份: 2016
• 资助国家: 日本
• 起止时间: 2016-10-07 至 2019-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16F16109/
• 关键词: sialidase; fluorescence; activation; cancer; self-immolative space; bacteria; Sialidase; Fluorescence activation; Lung cancer; Self-immolative space; Sialic acid substrate; Cancer imaging

Sialidases are members of a family of exoglycosidases specifically catalyzing the removal of sialic acid residues on the surface of exposed glycoconjugates. A novel fluorescence probe HMRef-S-Neu5Ac has been synthesised by incorporating a self-immolative spacer group between the sialic acid residue and the HMRef fluorophore to prevent hydrolysis via an acid-mediated pathway (i.e. in the absence of the target enzyme sialidase). First, kinetic measurements have shown the probe to have similar properties to that of a commercially available sialidase substrate, 4-MUNANA, demonstrating that the spacer group does not impede substrate recognition. Second, the stabilities of HMRef-S-Neu5Ac and a commercially avaiable 4MU-Neu5Ac were monitored over time at selected pHs of 2, 4, 6, 7.4 and 8. 4MU-Neu5Ac only showed significant instability at pH 2 where over the two-hour period fluorescence increased 5.6-fold. HMRef-S-Neu5Ac showed stability across all pHs within the two-hour experiment time. These experiments demonstrate that this probe is a better probe than 4MU-Neu5Ac due to its superior stability allowing for specificity for sialidase hydrolysis over acidic hydrolysis. The probe has been assessed in prostate cancer cells (LNCaP, DU145, PC3) which are reported to have significant levels of sialidase. Furthermore, some preliminary experiments of bacterial samples using this probe have been very positive and are currently under investigation.

唾液酸酶是外切糖苷酶家族的成员，专门催化去除暴露的糖复合物表面上的唾液酸残基。通过在唾液酸残基和 HMRef 荧光团之间掺入自毁间隔基团，合成了一种新型荧光探针 HMRef-S-Neu5Ac，以防止通过酸介导途径（即在不存在目标酶唾液酸酶的情况下）发生水解。首先，动力学测量表明该探针具有与市售唾液酸酶底物 4-MUNANA 相似的特性，证明间隔基团不会妨碍底物识别。其次，在选定的 pH 值 2、4、6、7.4 和 8 下，随时间监测 HMRef-S-Neu5Ac 和市售 4MU-Neu5Ac 的稳定性。4MU-Neu5Ac 仅在 pH 2 时表现出显着的不稳定性，而在这两种情况下，小时周期荧光增加5.6倍。 HMRef-S-Neu5Ac 在两小时的实验时间内在所有 pH 值下均表现出稳定性。这些实验表明，该探针是比 4MU-Neu5Ac 更好的探针，因为它具有优异的稳定性，允许唾液酸酶水解比酸性水解具有特异性。该探针已在前列腺癌细胞（LNCaP、DU145、PC3）中进行了评估，据报道这些细胞具有显着水平的唾液酸酶。此外，使用该探针对细菌样品进行的一些初步实验非常积极，目前正在调查中。

项目成果

シアリダーゼ活性検出蛍光プローブ

唾液酸酶活性检测荧光探针

• 发表时间: 2019

Development of a highly sensitive fluorescence probe for the detection of sialidase in cancer tissue

开发用于检测癌组织中唾液酸酶的高灵敏度荧光探针

• 发表时间: 2017
• 作者: Charlotte Rivas,Mako Kamiya;Yasuteru Urano
• 通讯作者: Yasuteru Urano