DL0805衍生物的生物活性筛选与细胞毒性评价  

Biological activity screening of DL0805 derivatives and the cytotoxicity evaluation

作　　者：袁天翊[1] 王夙博[1,2] 陈俞材 张惠芳[1] 焦晓臻[3] 谢平[3] 方莲花[1] 杜冠华[1] 

机构地区：[1]中国医学科学院&北京协和医学院药物研究所药物靶点研究与新药筛选北京市重点实验室,北京100050 [2]山东大学国家糖工程技术研究中心,济南250100 [3]中国医学科学院&北京协和医学院药物研究所活性物质发现与适药化研究北京市重点实验室,北京100050

出　　处：《中国新药杂志》2016年第19期2183-2188,共6页Chinese Journal of New Drugs

基　　金：国家自然科学基金面上项目(81573645);国家自然科学基金青年基金资助项目(81603101);国家"重大新药创制"科技重大专项资助项目(2013ZX09103001-008;2013ZX09508104;2013ZX09402203);药物研究所基本业务费(2016CX09)

摘　　要：目的:DL0805是本实验室发现的具有Rho激酶抑制作用的吲唑类化合物,本研究拟对DL0805结构改造后得到的105个小分子化合物进行体外活性筛选,发现潜在候选药物。方法:利用ELISA方法评价105个化合物的Rho激酶活性抑制作用;采用离体大鼠胸主动脉血管环张力检测系统评价化合物舒张血管作用;利用MTT法,检测化合物对血管平滑肌细胞(VSMC)和人脐静脉内皮细胞(HUVEC)的细胞毒性,并评价化合物对VSMC增殖的抑制作用。结果:经过初筛和复筛,发现5个化合物对Rho激酶活性具有较强的抑制作用;22个化合物对离体血管具有良好的舒张作用;1个化合物对VSMC毒性作用较强;5个化合物对血清诱导的VSMC增殖有一定的抑制作用;3个化合物对HUVEC毒性较强。结论:对DL0805衍生物的生物活性和细胞毒性的系统评价和综合分析,105个化合物中发现5个化合物具有良好的Rho激酶抑制作用、舒张血管活性及较低的细胞毒性,具有继续研究和开发的潜力,其中4个化合物为全新结构,值得进一步研究。Objective： To evaluate the in vitro activity of 105 derivatives of DL0805,a Rho-kinase inhibitor with indazole scaffold discovered in our previous work,and discover potential candidate compounds. Methods：Rho-kinase inhibitors among these 105 compounds were screened by ELISA and the vasorelaxant effects of these compounds on isolated rat thoracic aorta rings were evaluated. The toxicity tests were performed on two kinds of vascular cells including VSMC（ vascular smooth muscle cells） and HUVEC（ human umbilical vein endothelial cells）. Also the inhibitory effects on VSMC proliferation were tested by MTT assay. Results： After preliminary andsecondary screening,it was found that there were five compounds owning potent Rho-kinase inhibitory effect.Twenty-two compounds had vasodilatory effect on isolated aorta rings. Only one compound exhibited toxicity on VSMC and five compounds could inhibit the proliferation of VSMC induced by FBS. Meanwhile,three compounds were toxic on HUVEC. Conclusion： There are five compounds from the 105 DL0805 derivatives having the potential for further development. Among them,four compounds are new chemical entities,which deserve more investigation.

关 键 词：DL0805衍生物 RHO激酶 离体血管环 血管平滑肌细胞 人脐静脉内皮细胞 

分 类 号：R96[医药卫生—药理学]