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.

唾液酸酶是外糖苷酶家族的成员，特异性地催化了在暴露的糖缀合物表面上清除唾液酸残基。一种新型的荧光探针HMREF-S-NEU5AC已通过在唾液酸残基和HMREF荧光团之间合成一个自暴力的间隔组，从而通过酸介导的途径进行水解（即，在靶酶唾液酸酶的情况下）。首先，动力学测量结果表明探针具有与市售唾液酸酶底物4-munana相似的特性，表明间隔群不会阻碍底物识别。其次，随着时间的推移，HMREF-S-NEU5AC和商业上可用的4MU-NEU5AC的稳定性在2、4、6、7.4和8。4MU-NEU5AC的选定pHS中受到监测，在两个小时的荧光期间，在pH 2处仅显示出明显的不稳定。 HMREF-S-NEU5AC在两个小时的实验中均显示出所有pHS的稳定性。这些实验表明，由于其出色的稳定性允许在酸性水解上取得特异性，因此该探针比4MU-NEU5AC更好。该探针已在前列腺癌细胞（LNCAP，DU145，PC3）中进行了评估，这些探针据报道具有显着水平的唾液酸酶。此外，使用此探针的细菌样品进行了一些初步实验，目前正在研究中。

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