左娜娜创新答辩ppt-阿卡宁衍生物合成产物中乙酰胆碱酯酶

阿卡宁衍生物合成产物中乙酰胆碱酯酶抑制剂的筛选答辩人：左娜娜导师：马志玲副教授目录选题意义研究方法结果与讨论实验创新点Ⅰ选题意义成功筛选出乙酰胆碱酯酶抑制剂克服传统的有机定向合成的局限性乙酰胆碱酯酶抑制剂阿尔茨海默病（Alzheimer’s，AD）是一种多发生于老年人中的缓慢进行性、智能衰退性疾病，俗称老年性痴呆病。AD的关键性症状主要是由胆碱能功能障碍所引起的，治疗AD病的主要途径是寻找新的中枢拟胆碱药，乙酰胆碱酯酶抑制剂就是其中的一类治疗药物。乙酰胆碱在乙酰胆碱酯酶存在的条件下发生的水解反应如下：1.Acetylthiocholine+H2O→acetate+thiocholine2.Thiocholine+DTNB→2-nitrobenzoate-5-mercaptothiocholine+5-thio-2-nitrobenzoate（TNB）（yellow）BACK传统有机合成方法前导物合成分离提纯测定生物活性化学预试传统方法的局限性：1)只适合易分离体系；2)只注重主要合成产物的信息Ⅱ实验内容原料：β,β-二甲基丙烯酰阿卡宁乙酰胆碱酯酶抑制剂IC50的测定薄层色谱TLC液质联用LC-MS测定混合物是否具有抑制活力分离筛选具有抑制活力物质定性确定物质合成Ⅲ结果与讨论⒈研究体系以及合成路线（1）以2-氨基吡啶为例，说明可能的合成途径NNH2OHOHOOOOOHOHOOOOOHOHOOHOHOHOOOONHNNHNNHNOOOOHOHNNOOHOHOHNNOOHOHNN推测：可能有不止六个加成产物……其余从略（2）2-氨基苯并噻唑与原料的可能合成路径OHOHOOOOOHOHOOOOSNHSNHOHOHOOOOOHOHOOHSHNSNH2OOOOHOHNSOOHOHOHNSOOONS399推测：可能有六个加成产物⒉IC50的测定结果051015200.00150.00200.00250.00300.0035OD432抑制剂用量/μg051015200.00100.00150.00200.00250.00300.0035OD432抑制剂用量/μg图一2-氨基吡啶合成的抑制剂混合物的IC50测定图二2-氨基苯并噻唑合成的抑制剂混合物的IC50测定结论：从上面两图可知，两个抑制剂混合物体系都有抑制活力。⒊薄层色谱法测定抑制活力图三混合物经TLC的分离结果用薄层色谱法可以鉴别出具有抑制能力的化合物。结论:图四混合物有效抑制成分的筛选⒋应用高效液相色谱法分离抑制剂混合物min0246810121416mAU-8-6-4-20DAD1B,Sig=500,8Ref=600,100(ZNN\050519\05051902.D)2.0452.7165.90713.10214.04415.05715.658图五2-氨基吡啶合成混合物经HPLC分离的结果min051015202530mAU-8-6-4-202DAD1B,Sig=500,8Ref=600,100(ZNN\050519\05051905.D)2.1472.30412.64813.25119.70120.13120.50520.935图六2-氨基苯并噻唑合成混合物经HPLC分离的结果⒋应用高效液相色谱法分离抑制剂混合物⒌液相－质谱联用确定分子结构RT:0.00-21.110123456789101112131415161718192021Time(min)020000040000060000080000010000001200000140000016000001800000200000022000002400000260000028000003000000320000034000003600000uAU12.164.292.4511.6610.062.773.199.3510.275.342.258.245.4812.887.8513.7014.731.8417.8015.4818.8119.87NL:3.70E6ChannelAUVpyridine图七2-氨基吡啶合成抑制剂混合物的PDA总流图以2-氨基吡啶合成抑制剂混合物为例：pyridine#327RT:12.47AV:1SB:711.79-12.25NL:5.54E5T:-cESIFullms[150.00-1500.00]150200250300350400450500550600650700750800850900m/z050000100000150000200000250000300000350000400000450000500000550000Intensity463.2464.3361.2485.1848.3823.4269.2362.3567.1587.0657.1735.9792.6880.3712.1543.0518.3356.7413.2374.9289.4629.8685.5197.3268.3443.2176.7图八12.47min对应的质谱图，分子量463可能的结构式有：OOOOOHOHNHNpyridine#311RT:11.86AV:1NL:3.07E5T:-cESIFullms[150.00-1500.00]150200250300350400450500550600650700750800850900wavelength(nm)020000400006000080000100000120000140000160000180000200000220000240000260000280000300000Intensity461.23361.27462.24362.24393.23878.29876.97640.59677.05483.03341.03550.92898.66591.06816.02500.40768.19309.36701.76578.72440.45249.20174.89306.97224.26图九11.66min对应的质谱图，分子量461.23可能的结构式有：OOOOOONHNOOOOOOHNNpyridine#271RT:10.32AV:1NL:1.01E6T:-cESIFullms[150.00-1500.00]150200250300350400450500550600650700750wavelength(nm)0500001000001500002000002500003000003500004000004500005000005500006000006500007000007500008000008500009000009500001000000Intensity393.25309.22394.28477.14655.46415.31281.26517.04363.05755.11642.51584.48670.76723.04310.37553.25475.00610.57279.35223.19191.01图十10.32min对应的质谱图，分子量393可能的结构图为：OOHOHOHNNOOHOHHNONpyridine#249RT:9.48AV:1NL:3.35E5T:-cESIFullms[150.00-1500.00]150200250300350400450500550600650700750800850900wavelength(nm)020000400006000080000100000120000140000160000180000200000220000240000260000280000300000320000Intensity393.20809.20810.15415.15378.25765.84833.45689.39362.30668.15309.17881.57531.15768.51494.76423.19606.87724.93808.32562.63331.01272.18233.14189.02pyridine#113RT:4.28AV:1NL:4.02E5T:-cESIFullms[150.00-1500.00]150200250300350400450500550600650700750800850900wavelength(nm)020000400006000080000100000120000140000160000180000200000220000240000260000280000300000320000340000360000380000400000Intensity409.18443.26547.18391.09525.11339.24779.33453.27737.13513.13718.53352.94819.33253.29566.70639.05508.98311.18654.32843.12884.56202.12230.12pyridine#317RT:12.09AV:1NL:2.60E5T:-cESIFullms[150.00-1500.00]150200250300350400450500550600650700750800850900wavelength(nm)020000400006000080000100000120000140000160000180000200000220000240000Intensity461.20361.25483.05405.32777.09406.39583.09565.14745.87706.42822.71393.26496.59643.01584.21438.62562.46869.96333.16697.77730.69788.27542.87322.86271.11183.24249.05pyridine#323RT:12.32AV:1NL:2.81E5T:-cESIFullms[150.00-1500.00]150200250300350400450500550600650700750800850900wavelength(nm)020000400006000080000100000120000140000160000180000200000220000240000260000280000Intensity361.21463.16461.11877.18464.23844.91692.83497.06363.29405.63545.02809.28662.98589.51744.89383.06879.20343.32422.59641.09506.76710.70323.26895.55272.99183.36241.06结论：•通过应用薄层色谱法，可以分离各个物质，并确定具有抑制活力的物质；•通过高效液相色谱法，可以分离混合物；•通过液质联用方法，可以对各个物质进行定性分析。Ⅳ创新点合成产物的混合体系不经分离直接进行筛选，对难分离体系有很大的优越性对合成过程中的所有产物进行分析、筛选，从而获得更多有用信息应用薄层色谱法分离同分异构体，同时直接在薄层板上检测其生物活性致谢感谢导师马志玲副教授的悉心指导感谢吴京洪、凌连生等老师的关心感谢师兄师姐对我的鼓励和热心帮助感谢我的同学对我的照顾和支持