目的以水溶性磷酸盐柱[6]芳烃(phosphate salt pillar[6]arene,PP6A)作为芫花素的药物载体即主体分子,天然药芫花素为客体分子,构筑了一种新型的芫花素/PP6A(genkwanin/PP6A,GK/PP6A)超分子主客体包合物,对其进行表征和性能分析,进一步...目的以水溶性磷酸盐柱[6]芳烃(phosphate salt pillar[6]arene,PP6A)作为芫花素的药物载体即主体分子,天然药芫花素为客体分子,构筑了一种新型的芫花素/PP6A(genkwanin/PP6A,GK/PP6A)超分子主客体包合物,对其进行表征和性能分析,进一步对包合机制进行研究。方法采用饱和水溶液法制备芫花素与PP6A的超分子包合物;以包合物的包合率为指标,选用物质的量比、包合温度、包合时间和甲醇与水的体积比4个因素,每个因素3个水平,以L9(34)设计正交试验,对GK/PP6A包合物的制备工艺进行优化筛选。采用紫外可见吸收光谱法、傅里叶变换红外光谱、X射线粉末衍射、扫描电子显微镜和差示扫描量热等分析方法对芫花素和PP6A包合前后的理化性质进行研究分析;通过半经验分子轨道法、分子对接和核磁共振波谱法揭示了芫花素与PP6A最稳定的包合模式。结果GK/PP6A包合物的最优制备工艺为PP6A与芫花素的物质的量比为1∶1,包合温度为50℃,包合时间8 h,甲醇与超纯水体积比为1∶8,GK/PP6A包合物的包合率为89.93%;芫花素和PP6A以物质的量比1∶1形成超分子包合物,其络合常数为49488 L/mol;半经验分子轨道法与分子对接揭示的包合机制与二维核磁共振氢谱推断的主客体可能的包合模式相互吻合;芫花素包合前后的水溶解性和稳定性显著提高,其中水溶解性由3.5μg/mL提升至540.5μg/mL,约提高了155倍。结论PP6A是芫花素的良好宿主,芫花素与PP6A形成包合物后,芫花素的水溶性和稳定性得到显著提高,疏水作用和氢键作用为包合物形成的主要驱动力。展开更多
Enzymes containing 3′→5′exonuclease activities play an important role in various key cellular and physiological processes.The development of fluorescence biosensor is an efficient method to detecting enzyme activit...Enzymes containing 3′→5′exonuclease activities play an important role in various key cellular and physiological processes.The development of fluorescence biosensor is an efficient method to detecting enzyme activity.Herein,a fluorescence resonance energy transfer(FRET)“on”and“off”strategy for detecting exonuclease III(Exo III)activity has been developed.We report here that the double-stranded DNA(dsDNA)enables to bind tightly to self-assembled nanosheets of cationic perylene monoimide derivative(PMI-O7)through electrostatic interaction,and the 6-carboxyfluorescein(FAM)-modified dsDNA could be efficiently quenched via FRET between FAM and PMI-O7.Upon the addition of Exo III,the dsDNA will be digested and the FAM fluorophore will be released,resulting in the fluorescence recovery of FAM.This method provides a simple and sensitive biosensor platform with a low detection limit of 0.077 U/mL for Exo III.Importantly,this method exhibits similar and calibration curves for the detection of Exo III in both buffer and fetal bovine serum samples,indicating that this platform has potential to detect Exo III activity in complex samples.展开更多
文摘目的以水溶性磷酸盐柱[6]芳烃(phosphate salt pillar[6]arene,PP6A)作为芫花素的药物载体即主体分子,天然药芫花素为客体分子,构筑了一种新型的芫花素/PP6A(genkwanin/PP6A,GK/PP6A)超分子主客体包合物,对其进行表征和性能分析,进一步对包合机制进行研究。方法采用饱和水溶液法制备芫花素与PP6A的超分子包合物;以包合物的包合率为指标,选用物质的量比、包合温度、包合时间和甲醇与水的体积比4个因素,每个因素3个水平,以L9(34)设计正交试验,对GK/PP6A包合物的制备工艺进行优化筛选。采用紫外可见吸收光谱法、傅里叶变换红外光谱、X射线粉末衍射、扫描电子显微镜和差示扫描量热等分析方法对芫花素和PP6A包合前后的理化性质进行研究分析;通过半经验分子轨道法、分子对接和核磁共振波谱法揭示了芫花素与PP6A最稳定的包合模式。结果GK/PP6A包合物的最优制备工艺为PP6A与芫花素的物质的量比为1∶1,包合温度为50℃,包合时间8 h,甲醇与超纯水体积比为1∶8,GK/PP6A包合物的包合率为89.93%;芫花素和PP6A以物质的量比1∶1形成超分子包合物,其络合常数为49488 L/mol;半经验分子轨道法与分子对接揭示的包合机制与二维核磁共振氢谱推断的主客体可能的包合模式相互吻合;芫花素包合前后的水溶解性和稳定性显著提高,其中水溶解性由3.5μg/mL提升至540.5μg/mL,约提高了155倍。结论PP6A是芫花素的良好宿主,芫花素与PP6A形成包合物后,芫花素的水溶性和稳定性得到显著提高,疏水作用和氢键作用为包合物形成的主要驱动力。
基金supported by the Natural Science Foundation of Guangdong Province,China (No.2014A030313194)the Science and Technology Program of Guangzhou,China (No.201804010017)+1 种基金the Science and Technology Program of Guangdong Province,China (No.2017A050506021)the National Natural Science Foundation of China (No.21374137).
文摘Enzymes containing 3′→5′exonuclease activities play an important role in various key cellular and physiological processes.The development of fluorescence biosensor is an efficient method to detecting enzyme activity.Herein,a fluorescence resonance energy transfer(FRET)“on”and“off”strategy for detecting exonuclease III(Exo III)activity has been developed.We report here that the double-stranded DNA(dsDNA)enables to bind tightly to self-assembled nanosheets of cationic perylene monoimide derivative(PMI-O7)through electrostatic interaction,and the 6-carboxyfluorescein(FAM)-modified dsDNA could be efficiently quenched via FRET between FAM and PMI-O7.Upon the addition of Exo III,the dsDNA will be digested and the FAM fluorophore will be released,resulting in the fluorescence recovery of FAM.This method provides a simple and sensitive biosensor platform with a low detection limit of 0.077 U/mL for Exo III.Importantly,this method exhibits similar and calibration curves for the detection of Exo III in both buffer and fetal bovine serum samples,indicating that this platform has potential to detect Exo III activity in complex samples.
