【作者】 李素芳； 刘飞； 刘巍； 赵悦； 吕志珍； 徐明； 张幼怡；

【Author】 LI Su-fang~1,LIU Fei~1,LIU Wei~2,ZHAO Yue~2,LU Zhi-zhen~1,XU Ming~1,ZHANG You-yi~1 (1.Key Laboratory of Molecular Cardiovascular Sciences of Ministry of Education,Key Laboratory of Cardiovascular Molecular Biology and Regulatory Peptides,Ministry of Health Institute of Vascular Medicine,Peking University Third Hospital,Beijing 100191,China;2.Department of Chemical Biology,College of Chemistry and Molecular Engineering,Peking University,Beijing 100871,China)

【机构】 北京大学第三医院血管医学研究所分子心血管教育部重点实验室,卫生部心血管分子生物学与调节肽重点实验室； 北京大学化学与分子工程学院化学生物学系；

【摘要】 目的检测一批新合成的α₁-肾上腺素受体（α₁-AR）拮抗剂对α₁-AR的选择性拮抗活性。方法①通过吲哚哌啶与哌嗪分子耦合衍生,得到一系列α₁-肾上腺素受体拮抗剂分子,化合物B1～B9,分别具有吲哚哌啶基和不同的取代基团。②应用离体大鼠左心耳收缩功能实验,检测IPD,化合物B1～B9对PE刺激下离体大鼠左心耳上α₁-AR的拮抗活性。③采用Western印迹法检测IPD,化合物B1～B9对PE刺激下293细胞内细胞外信号调节激酶（ERK）磷酸化水平的影响。结果①成功合成了具有吲哚哌啶基和不同取代基团的潜在α₁-AR拮抗剂。②提前孵育α₁-AR拮抗剂酚妥拉明或IPD,化合物B1,B3,B4,B7,B8,B9,PE引起的离体大鼠左心耳的收缩反应均被有效抑制;其中IPD,化合物B4和B8引起收缩曲线的明显右移,IPD,化合物B4和B8的pA₂值分别是6.72±0.21,6.86±0.29和6.67±0.19。③在稳定表达α_1A-AR的HEK293细胞内,化合物B1,B2,B3,B5,B6,B7,B8,B9或IPD均较明显地抑制PE引起的ERK1/2的磷酸化增强;在稳定表达α_1B-AR的HEK293细胞内,化合物B2,B4,B7或B8较明显地抑制PE引起的ERK1/2的磷酸化增强。结论化合物B4能够选择性地拮抗α_1B-AR的活性;化合物B1,B3,B5,B6,B9和IPD能够选择性地拮抗α_1A-AR的活性。更多还原

【Abstract】 OBJECTIVE To investigate the blocking activities of a series of potentialα₁-adrenoceptor（α₁-AR） antagonists（Compounds Bl -B9） onα₁-AR.METHODS CD A series of potentialα₁-adrenoceptor（α₁-AR） antagonists,indolylpiperidine derivative（IPD） and Compounds Bl - B9,with indolylpiperidine moiety and different substitutes were synthesized through the coupling of indolylpiperidine and piperazine derivatives.（2） Inotropic responses experiment was used to examine blocking effects of IPD and Compounds Bl - B9 in isolated rat atria by phenylephrine（PE） stimulation.③Blocking effect of IPD and Compounds Bl - B9 on phosphorylation level of extracellular signal-regulated kinase（ERK） in PE treated HEK293 cells was tested by Western blotting.RESULTS （I） Potential a,-adrenoceptor（α₁-AR） antagonists with indolylpiperidine moiety and different substitutes were synthesized successfully.（2） PE caused a dose-dependent inotropic response which was inhibited by pre-incubation of phentolamine（Phen）,a non-selectiveα₁-AR antagonist,IPD and Compounds Bl,B3,B4,B7,B8 and B9,respectively; IPD and Compounds B4 and B8 caused an obvious rightward shift of inotropic response-curve,the pA₂ values for IPD and Compounds B4 and B8 were 6.72±0.21,6.86±0.29 and 6.67±0.19,respectively.③Phosphorylation level of ERK1/2 was inhibited by pre-incubation with Compounds B1,B2,B3,B5,B6,B7,B8 and B9 or IPD in PE treatedα_1A-AR stably expressed HEK293 cells;PE-stimulated phosphorylation level of ERK1/2 was inhibited by pre-incubation with Compounds B2,B4,B7 or B8 inα_1B-AR stably expressed HEK293 cells.CONCLUSION Compound B4 has a selective blocking activity onα_1B-AR,and Compounds B1,B3,B5, B6 and B9 or IPD have a selective blocking activity on the phosphorylation level of ERK1/2.更多还原