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Control of the drug efflux transport from the brain utilizing the transport function for the large molecules〜

利用大分子的转运功能控制药物从大脑的流出转运〜

基本信息

批准号：
11672270
负责人：
DEGUCHI Yoshiharu
金额：
$ 2.18万
依托单位：
Hokkaido College of Pharmacy (2000)University of Shizuoka (1999)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

项目摘要

The final goal of this study is to control the expression of drug efflux transporter in the blood-brain barrier (BBB) and to improve the brain transferability of the drug. There is the possibility of suppressing the efflux transporter by introducing the antibody and/or antisense gene in the brain capillary endothelial cell. However, these giant molecule does not BBB easily. Some bioactive peptides and proteins are exceptionally taken up by the brain through the specialized transportation system of the BBB.If such transport system is utilized, antisense gene may be delivered into the brain capillary endothelial cells. However, information on the peptide/or protein transport system in the BBB is limited, In this study, the researcher clarified for the first time that heparan sulfate proteoglycan (HSPG) of the BBB functions transport system of large molecules including bioactive peptides and proteins.Using conditionally immortalized mouse brain capillary endothelial cell lines (TM-EBB), i … More t was confirmed that the gene expression of mouse perlecan which is the core protein of HSPG is expressed at TM-BBB.Next, the transport function of HSPG is examined using ^<125>l-bFGF as a ligand to HSPG.^<125>l-bFGF avidly bound to the brain capillary isolated from the bovine brain. Internalization of ^<125>l-bFGF into TM-BBB showed concentration-dependence, temperature and osmolarity dependence. In addition, the internalization was significantly inhibited in heparin, chondroitin sulfate B, and it was also inhibited in cells treated with heparinase and sodium chlorate, and antibody to mouse perlecan. In situ brain perfusion of ^<125>l-bFGF revealed the uptake into the brain parenchyma of unchanged ^<125>l-bFGF.This brain uptake of ^<125>l-bFGF was significantly inhibited by the heparin co-perfusion. From these results, it became clear that HSPG is expressed in the BBB and that it functions as the transport system of large molecule to the brain. By utilizing this transport system in future, antibody and/or antisense gene, which controls the function of the efflux transporter in the BBB, may be efficiently delivered into the brain endothelial cells. Less

本研究的最终目的是控制药物外排转运体在血脑屏障（BBB）中的表达，提高药物的脑转移性。存在通过在脑毛细血管内皮细胞中引入抗体和/或反义基因来抑制外排转运蛋白的可能性。然而，这些大分子并不容易BBB。某些生物活性肽和蛋白质通过血脑屏障的特殊转运系统被脑摄取，利用这种转运系统，可以将反义基因导入脑毛细血管内皮细胞。本研究首次阐明了血脑屏障中硫酸乙酰肝素蛋白聚糖（heparan sulfate proteoglycan，HSPG）在生物活性肽和蛋白质等大分子物质的转运系统中的作用。利用条件永生化小鼠脑毛细血管内皮细胞系（conditionally immortalized mouse brain capillary endothelial cell lines，TM-EBB），通过体外培养，在体外培养条件永生化小鼠脑毛细血管内皮细胞系（conditionally immortalized mouse brain capillary endothelial cell lines，TM-EBB），在体外培养条件永生化小鼠脑毛细血管内皮细胞系（conditionally immortalized mouse brain capillary endothelial cell lines，TM-EBB），在体外培养条件永生化小鼠脑毛细血管内皮细胞系（conditionally immortalized mouse brain capillary endothelial cell lines，TM-EBB），在体外培养条件永生化小鼠脑毛细血管内皮细胞系（conditionally immortalized mortalized mouse brain ...更多信息证实了HSPG的核心蛋白--小鼠串珠素的基因表达在TM-BBB中表达。接着，使用β 1-bFGF作为HSPG的配体，检查HSPG的转运功能。<125><125>L-bFGF与从牛脑分离的脑毛细血管紧密结合。TM-BBB对bFGF的内化具有浓度依赖性、温度依赖性和渗透压依赖性。<125>此外，肝素、硫酸软骨素B、肝素酶、氯酸钠和抗小鼠串珠素抗体处理的细胞也能显著抑制内化。在体脑灌注显示，脑实质中摄取了未改变的^l-bFGF，这种脑摄取被肝素共灌注显著抑制。<125><125><125>从这些结果可以清楚地看出，HSPG在BBB中表达，并且它作为大分子到脑的运输系统发挥作用。通过将来利用这种转运系统，可以将控制BBB中外排转运蛋白功能的抗体和/或反义基因有效地递送到脑内皮细胞中。少