新穎螢光葡萄糖類似物之特性研究與應用

Characterization of a novel fluorescent glucose derivative and its applications

高廷宇(Ting-Yu Kao)

指導教授 ： 許麗卿

國立臺灣大學/醫學院/藥學研究所/碩士(2014年)

https://doi.org/10.6342/NTU.2014.10636

摘要

葡萄糖對細胞來說是提供能量的重要來源，細胞主要是以兩種轉運蛋白將葡萄糖送入細胞內，一種是透過sodium-glucose co-transporters (以下簡稱 SGLT) 以主動運輸的方式，而另外一種是透過glucose transportes (以下簡稱 GLUT ) 以被動運輸的方式將葡萄糖送入細胞中。因此若能夠以簡單的方法得知葡萄糖運送入細胞的狀態，便能夠成為一個有效的研究工具。
1-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxy-D-glucose
(以下簡稱為 1-NBDG ) 是一種新合成的螢光標定葡萄糖，它可以透過葡萄糖轉運蛋白進入細胞中，達到標定細胞的目的。和 2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxy-D-glucose (以下簡稱為 2-NBDG ) 相比，1-NBDG是更好的SGLT受質，能有效地被SGLT運送進入細胞。但是對於其他的特性尚有許多不了解的地方，因此本文會對其物質的安定性、在細胞中的代謝情況作探討。並嘗試使用流式細胞儀來偵測標定的細胞，以提高運用的廣泛度。最後則是利用1-NBDG所建立的SGLT1和SGLT2抑制劑細胞篩選平台對天然物的萃取物進行篩選，得到可能的前導性藥物。

關鍵字

1-NBDG ； 2-NBDG ； 選擇性 SGLT2 抑制劑 ； 流式細胞儀 ； 第二型糖尿病

並列摘要

Glucose is an important energy source for cells. Cells transport glucose by two types of glucose transporters: the active transporters sodium-glucose co-transporters (SGLT), and the passive transporters glucose transportes (GLUT) . If we can develop an easy way to detect glucose in the cell, it could be a useful tool for research.
1-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxy-D-glucose (1-NBDG) is a newly developed fluorescence-labeled glucose. It can be transported into cells through glucose transporters. Compared with -(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxy-D-glucose (2-NBDG), 1-NBDG can be transported by SGLTs more effectively. 1-NBDG is not fully characterized yet. Therefore, in this thesis, we investigate the stability of 1-NBDG, determine its intracellular metabolites, and set up a flow cytometric method for the detection of 1-NBDG uptake in cells. Furthermore, 1-NBDG is also used in a high-throughput cell-based method to screen for potential SGLT1 and SGLT2 inhibitors from natural products.

並列關鍵字

1-NBDG ； 2-NBDG ； selective SGLT2 inhibitor ； Type2 diabetes mellitus ； flow cytometry

參考文獻

1. Idris I and Donnelly R., Sodium-glucose co-transporter-2 inhibitors: an emerging new class of oral antidiabetic drug. Diabetes Obes Metab, 2009. 11(2): p 79-88.

2. Kinne RK and Castaneda F., SGLT inhibitors as new therapeutic tools in the treatment of diabetes. Handb Exp Pharmacol, 2011(203): p 105-26.

3. Washburn WN., Evolution of sodium glucose co-transporter 2 inhibitors as anti-diabetic agents. Expert Opin Ther Pat, 2009. 19(11): p 1485-99.

4. Freitas HS, Anhe GF, Melo KF, Okamoto MM, Oliveira-Souza M, Bordin S and Machado UF., Na(+) -glucose transporter-2 messenger ribonucleic acid expression in kidney of diabetic rats correlates with glycemic levels: involvement of hepatocyte nuclear factor-1alpha expression and activity. Endocrinology, 2008. 149(2): p 717-24.

5. Vallon V., Molecular determinants of renal glucose reabsorption. Focus on "Glucose transport by human renal Na+/D-glucose cotransporters SGLT1 and SGLT2". Am J Physiol Cell Physiol, 2011. 300(1): p C6-8.