131I标记热休克蛋白90抑制剂17-丙烯胺基-17-去甲氧基格尔德霉素的方法学研究-东南大学学报医学版

131I标记热休克蛋白90抑制剂17-丙烯胺基-17-去甲氧基格尔德霉素的方法学研究

单位：东南大学,核医学技术研究所,江苏,南京,210009

关键词：131碘 17-丙烯胺基-17-去甲氧基格尔德霉素热休克蛋白90 放射性同位素生物分布

分类号：R730.55

出版年·卷·期（页码）：2007·26·第三期（170-173）

摘要：

目的:建立131I标记热休克蛋白90抑制剂17-丙烯胺基-17-去甲氧基格尔德霉素(17-AAG)的方法,探讨其标记条件.方法:采用放射性碘标记氯胺-T法标记17-AAG,并研究其在ICR正常小鼠体内的生物分布.结果:131I氯胺-T法标记17-AAG 的最佳条件为:17-AAG乙醇溶液40μl(含17-AAG 5μg),0.5mol·L-1磷酸盐缓冲液50μl(pH 7.5),Na 131I溶液20μl,氯胺-T 30μg·(15μl)-1(用pH 7.5的0.05mol·L-1 PB溶解),25℃反应50s,偏重亚硫酸钠40μg·(20μl)-1(用pH 7.5的0.05mol·L-1 PB溶解)中止反应.标记率约为53.1%,经乙酸乙酯纯化后所得标记物的放射化学纯度>90%;其生理盐水溶液4℃放置5d,放射化学纯度大于90%.小鼠尾静脉注射131I-17-AAG后0.5h 131I-17-AAG在胆囊达到高峰(369.44±147.81) %ID·g-1,肝脏中仅有(2.23±0.50)%ID·g-1,说明对肝脏毒性小.结论:热休克蛋白90抑制剂17-AAG的标记方法操作简便,反应时间短.

Objective To study the method for labeling 17-allylamino,17-demethoxygeldanamycin(17-AAG) with 131I and its biodistribution in normal mice.Methods 131I17-AAG was prepared by the reaction of 17AAG with Na 131I in the presence of chloramines-T.The conditions of labeling,the stability of 131I-17AAG and the biodistribution in normal mice were investigated.Results To search for the optimal conditions,5μg 17AAG dissolved by 40μl alcohol,50μl 0.5mol·L-1 phosphate buffer saline(pH 7.5),20μl Na 131I(185 MBq·ml-1),chloramines-T(2 g·L-1) dissolved by 0.05mol·L-1 phosphate buffer(pH 7.5) were mixed at 25 ℃ for 50 seconds.After the labeling reaction,an excess of sodium metabisulfite dissolved by 0.05mol·L-1 phosphate buffer(pH 7.5) was added to terminate the reaction.The average labeling efficiency was around 53.1%.The radiochemical purity of 131I-17-AAG was over 90% after purification.131I-17-AAG showed the stability above 90% in saline up to 5d at 4 ℃.The biodistribution study in normal mice showed that there was high uptake in cholecyst,stomach and intestine.Conclusion Our preliminary results demonstrate that the labeling of 131I-17-AAG is simple and successful.Further study is going on.

参考文献：

[1] GOETZ M P, TOFT D O, AMES M M. The Hsp90 chaperone complex as a novel target for cancer therapy. 2003(8). doi:10.1093/annonc/mdg316
[2] KUMAR V, GREEN S, STAUB A. Localization of the estradiol binding and putative DNA binding domains of the human oestrogen receptor, 1986(9)
[3] GROSSMANN M E, HUANG H, TINDALL D. Androgen receptor signalling in androgen refractory prostate cancer. 2001(22). doi:10.1093/jnci/93.22.1687
[4] MA Y Y, WEI S J, LIN Y. PI3CA as an oncogene in cervical cancer. 2000(23). doi:10.1038/sj.onc.1203597
[5] BEARSS D J, HURLEY L H, von HOFF D D. Telomere maintenance mechanisms as a target for drug development. 2000(56). doi:10.1038/sj.onc.1204092
[6] XU W, MIMNUAUGH E G, KIM J S. Hsp90,not Grp94,regulates the intracellular trafficking and stability of nascent ErbB2, 2002(1)
[7] KAMAL A, THAO L, SENSINTAFFAR J. A high affinity conformation of Hsp90 confers tumour selectivity on Hsp90 inhibitors. 2003(6956). doi:10.1038/nature01913

服务与反馈：

【文章下载】【发表评论】【查看评论】【加入收藏】

提示：您还未登录，请登录！点此登录