CN110256413A - 一种新型aie荧光探针分子及该探针分子、纳米胶束和囊泡的制备方法 - Google Patents
一种新型aie荧光探针分子及该探针分子、纳米胶束和囊泡的制备方法 Download PDFInfo
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Abstract
本发明涉及聚集诱导发光效应和纳米探针领域，具体为一种新型AIE荧光探针分子及该探针分子、纳米胶束和囊泡的制备方法。本发明通过化学合成得到两亲性AIE分子，并可自组装成生物相容性好、稳定性高的纳米胶束或囊泡。通过本发明得到的AIE分子具有羟基、季铵盐和羧基的组成亲水性头，AIE性质的多个苯环和柔性十六烷基烃组成疏水性尾，利用特定的组装条件可组装形成纳米胶束或囊泡，且具有良好的稳定性。本发明纳米胶束和囊泡制备过程简单，可重复性高。
Description
技术领域
本发明涉及聚集诱导发光效应和纳米探针领域，具体为一种新型AIE荧光探针分子及该探针分子、纳米胶束和囊泡的制备方法。
背景技术
随着聚集诱导发光(aggregation-inducedemission)现象的发现，科研工作者已经合成了许多具有不同性质的AIE化合物。AIE材料在化学、材料和生命科学等领域均具有广泛的应用，已涵盖生物探针、化学传感器、光电子器件以及刺激响应材料等。AIE分子解决了常见荧光分子聚集导致荧光淬灭、光稳定性差等缺点，并且具有良好的生物相容性，因此在生物医疗领域作为荧光探针具有很大的应用前景。
常见AIE分子为疏水性分子，可通过纳米沉淀法得到纳米点，但其稳定性差，易聚集成较大的微米颗粒。纳米胶束和囊泡均具有较高的稳定性，在纳米材料领域已广泛应用。把AIE分子进行结构修饰，通过特定组装条件则有可能得到具有AIE性质的纳米胶束或囊泡。
综上所述，本发明通过三苯基胺为AIE母核引入亲水性头部和柔性疏水性十六烷基烃链，得到两亲性的AIE小分子，并自组装成纳米胶束或囊泡，并且具有良好的生物相容性、可生物降解性、光稳定性等优良性能。可作为载体在药物递送、基因递送、光动力治疗等领域具有很大的应用前景。
发明内容
本发明所要解决的技术问题是：同一个两亲性AIE分子可通过不同的组装条件可能到纳米胶束和囊泡目前尚未见报道，本发明为填补研究空白，通过化学合成得到两亲性AIE分子，并可自组装成生物相容性好、稳定性高的纳米胶束或囊泡。
为了实现上述目的，苯发明是通过以下技术方案实现的：一种新型AIE荧光探针分子，其结构式如下：
本发明的第二个技术方案是：一种新型AIE荧光探针分子,其合成路线如下：
本发明的第三个技术方案是：一种新型AIE荧光探针分子,其合成步骤如下：
1)对碘苯酚和碘代十六烷溶于DMF中，加入碳酸钾，室温反应过夜，后处理，得到化合物a；
2)化合物a和对溴苯胺溶于甲苯中，加入氢氧化钾，在1,10-菲咯啉和碘化亚铜催化下得到化合物b；
3)化合物b和5-醛基2-噻吩硼酸溶于甲醇和甲苯(v/v＝1:1)溶剂中，加入碳酸钾，在双三苯基膦二氯化钯催化下，60-90℃反应12-24h，后处理得到化合物c；
4)化合物c和对乙酸吡啶盐酸盐溶于氯仿中，加入哌啶为催化剂，60-80℃反应过夜，后处理得到化合物APN；
5)化合物APN溶于氯仿中，加入2-溴乙醇，60-80℃反应过夜，后处理得到AIE探针分子APNO。
本发明的第四个技术方案是：一种新型AIE荧光探针分子的纳米胶束和囊泡的制备方法，包括如下步骤：
1)AIE探针分子APNO溶于四氢呋喃中，加入对应摩尔比的胆固醇:
其中,APNO/胆固醇的摩尔比范围为1:0-1:2；
2)磁力搅拌下，用注射器缓慢滴加1-3mL去离子水，室温搅拌0.5-2h；
3)用去离子水透析24-72h，每两小时换一次水，得到纳米胶束或囊泡。
有益效果
1.纳米胶束和囊泡制备过程简单，可重复性高；
2.组装纳米粒径均一，并且平均粒径在200纳米左右，静脉注射后，可通过实体肿瘤的EPR效应，到达被动靶向肿瘤组织，减少对正常组织的毒副作用；
3.可作为其他药物载体，递送药物至肿瘤部位，也可本身作为治疗试剂；
4.聚集诱导发光探针分子APNO本身可发射荧光，可用于肿瘤的诊断；
5.聚集诱导发光探针分子APNO可在白光照射下产生活性氧，当APNO纳米颗粒到达肿瘤后，在光照下产生活性氧，杀死肿瘤细胞，用于肿瘤的光动力治疗。
附图说明
图1为AIE荧光探针分子APNO的紫外吸收和荧光发射谱图。
图2为APNO在不同体积水/THF(ft)中的荧光发射谱图。
图3为实施例1的动态光散射图。
图4为实施例1的透射电镜图。
图5为实施例3的动态光散射图。
图6为实施例3的透射电镜图。
具体实施方式
以下结合附图和实施例来对本发明做进一步的说明。应当理解的是，这里所讨论的实施例只是为了说明，对本领域技术人员来说，可以加以改进或变换，而所有这些改进和变换都应属于本发明所附权利要求的保护范围。
实施例1
将对碘苯酚(10g，45.4mmol)和碘代十六烷(16g，45.4mmol)溶于200ml DMF中，室温反应过夜。加入400mL乙酸乙酯，分别用1M的氢氧化钠水溶液200ml洗三次，饱和食盐水洗一次有机相。有机相无水硫酸镁干燥，过滤，旋转蒸发除溶剂，硅胶柱纯化，洗脱剂为石油醚：乙酸乙酯＝500:1，得到产物a，重量17.69g，产率：87.6％。1HNMR(400MHz，CDCl3)δ7.53(d,J＝12Hz,2H),6.67(d,J＝8Hz,2H),3.90(t,J＝4Hz,2H),1.80-1.71(m,2H),1.48-1.38(m,2H),1.36-1.23(m,24H),0.88(t,J＝8Hz,3H)。
氩气保护下，在反应瓶中一次加入化合物a(9.98g，22.46mmol)，对溴苯胺(1.68g，9.76mmol)，氢氧化钾(3.83g，68.32mmol)，1,10-菲咯啉(36mg，0.2mmol)，碘化亚铜(38mg，0.2mmol)和70mL甲苯，120℃回流反应24h。减压除溶剂，硅胶柱纯化，洗脱剂为石油醚：乙酸乙酯＝6:1，得到产物b，重量1.35g，产率为16.7％。1HNMR(400MHz，CDCl3)δ7.22(d,J＝8Hz,2H),7.01(d,J＝12Hz,4H),6.83-6.76(m,6H),3.91(t,J＝8Hz,4H),1.80-1.72(m,4H),1.50-1.40(m,4H),1.31-1.24(m,48H),0.91-0.85(t,J＝8Hz,6H)。
20ml无水THF中加入b(1.88g，2.30mmol)、5-甲酰-2噻吩硼酸(0.74g，4.60mmol)、Pd(PPh3Cl)2(0.16g，0.23mmol)、碳酸钾(1.59g，11.48mmol)，氩气保护，70℃反应过夜，减压除溶剂，加入40mL乙酸乙酯，分别用30mL蒸馏水萃取三次和30mL饱和食盐水萃取一次有机相，有机相无水硫酸镁干燥，过滤，硅胶柱纯化，洗脱剂为二氯甲烷：石油醚＝2:1，得到产物c，重量0.48g，产率为25％。1HNMR(400MHz，CDCl3)δ9.83(s,1H),7.68(d,J＝4Hz,1H),7.45(d,J＝8Hz,2H),7.25(d,J＝4Hz,1H),7.07(d,J＝8Hz,4H),6.92-6.80(m,6H),3.94(t,J＝8Hz,4H),1.80-1.72(m,4H),1.50-1.41(m,4H),1.32-1.24(m,48H),0.88(t,J＝8Hz,6H)。
反应瓶中依次加入c(1.8g，2.15mmol)，对乙酸吡啶盐酸盐(1.12g，6.46mmol)，哌啶(0.91g，10.75mmol)和40mL氯仿，70℃反应24h，调pH至6.0分别用40mL水和40mL饱和食盐水萃取有机相，无水硫酸钠干燥，硅胶柱纯化，洗脱剂为二氯甲烷：甲醇＝20:1,，得到产物APN，重量0.25g，产率12.1％。1HNMR(400MHz，CDCl3)δ8.57(s,1H),7.43-7.26(m,6H),7.12-7.01(m,6H),6.90(d,J＝8Hz,2H),6.83(d,J＝8Hz,4H),6.74(d,J＝12Hz,1H),3.92(t,J＝8Hz,4H),1.82-1.72(m,4H),1.49-1.40(m,4H),1.35-1.21(m,48H),0.88(t,J＝8Hz,6H)。
反应瓶中依次加入化合物APN(150mg，0.16mmol)，2-溴乙醇(98mg，0.78mmol)和5mL氯仿，70℃反应48h，减压除溶剂，制备硅胶层析柱纯化，展开剂为二氯甲烷：甲醇＝8:1，得到产物APNO 140mg，产率为81.4％。1HNMR(400MHz，d6-DMSO)δ8.79(d,J＝8Hz,2H),8.16(d,J＝8Hz,2H),7.53(d,J＝8Hz,2H),7.48(d,J＝4Hz,1H),7.44(d,J＝4Hz,1H),7.13-7.03(m,5H),6.93(d,J＝8Hz,4H),6.76(d,J＝8Hz,2H),4.51(t,J＝4Hz,2H),3.94(t,J＝8Hz,4H),3.84(t,J＝8Hz,2H),1.75-1.65(m,4H),1.43-1.38(m,4H),1.29-1.21(m,48H),0.85(t,J＝8Hz,6H)。
1.2mgAIE探针分子APNO溶于3mL四氢呋喃中，不加入胆固醇；磁力搅拌下，用注射器缓慢滴加3mL去离子水，APNO最终浓度为0.2mg/mL，室温搅拌1h；得到的溶液转移至透析袋(截留分子量8000-14000)中，用去离子水透析24h，每两小时换一次水，得到纳米胶束。动态光散射仪测试得平均粒径为230nm，PDI为0.149。
实施例2
1.2mg AIE探针分子APNO溶于3mL四氢呋喃中，加入0.22mg胆固醇；磁力搅拌下，用注射器缓慢滴加3mL去离子水，APNO最终浓度为0.2mg/mL，室温搅拌1h；得到的溶液转移至透析袋(截留分子量8000-14000)中，用去离子水透析24h，每两小时换一次水，得到纳米胶束。动态光散射仪测试得平均粒径为258nm，PDI为0.192。
实施例3
1.2mg AIE探针分子APNO溶于3mL四氢呋喃中，加入0.55mg胆固醇；磁力搅拌下，用注射器缓慢滴加3mL去离子水，APNO最终浓度为0.2mg/mL，室温搅拌1h；得到的溶液转移至透析袋(截留分子量8000-14000)中，用去离子水透析24h，每两小时换一次水，得到纳米胶束。动态光散射仪测试得平均粒径为251nm，PDI为0.222。
实施例4
1.2mg AIE探针分子APNO溶于3mL四氢呋喃中，加入0.87mg胆固醇；磁力搅拌下，用注射器缓慢滴加3mL去离子水，APNO最终浓度为0.2mg/mL，室温搅拌1h；得到的溶液转移至透析袋(截留分子量8000-14000)中，用去离子水透析24h，每两小时换一次水，得到纳米胶束。动态光散射仪测试得平均粒径为209nm，PDI为0.122。
实施例5
化合物a和b的合成过程同实施例1。
无水THF中加入b、5-甲酰-2噻吩硼酸、Pd(PPh3Cl)2、碳酸钾，氩气保护，60℃反应24h，减压除溶剂，加入乙酸乙酯，分别用蒸馏水萃取三次和饱和食盐水萃取一次有机相，有机相无水硫酸镁干燥，过滤，硅胶柱纯化，得到产物c。
反应瓶中依次加入c，对乙酸吡啶盐酸盐，哌啶和氯仿，60℃反应24h，调pH至6.0分别用水和饱和食盐水萃取有机相，无水硫酸钠干燥，硅胶柱纯化，得到产物APN。
反应瓶中依次加入化合物APN，2-溴乙醇和氯仿，60℃反应48h，减压除溶剂，制备硅胶层析柱纯化，得到产物APNO。
实施例6
化合物a和b的合成过程同实施例1。
无水THF中加入b、5-甲酰-2噻吩硼酸、Pd(PPh3Cl)2、碳酸钾，氩气保护，75℃反应16h，减压除溶剂，加入乙酸乙酯，分别用蒸馏水萃取三次和饱和食盐水萃取一次有机相，有机相无水硫酸镁干燥，过滤，硅胶柱纯化，得到产物c。
反应瓶中依次加入c，对乙酸吡啶盐酸盐，哌啶和氯仿，78℃反应24h，调pH至6.0分别用水和饱和食盐水萃取有机相，无水硫酸钠干燥，硅胶柱纯化，得到产物APN。
反应瓶中依次加入化合物APN，2-溴乙醇和氯仿，75℃反应48h，减压除溶剂，制备硅胶层析柱纯化，得到产物APNO。
实施例7
化合物a和b的合成过程同实施例1。
无水THF中加入b、5-甲酰-2噻吩硼酸、Pd(PPh3Cl)2、碳酸钾，氩气保护，90℃反应12h，减压除溶剂，加入乙酸乙酯，分别用蒸馏水萃取三次和饱和食盐水萃取一次有机相，有机相无水硫酸镁干燥，过滤，硅胶柱纯化，得到产物c。
反应瓶中依次加入c，对乙酸吡啶盐酸盐，哌啶和氯仿，80℃反应24h，调pH至6.0分别用水和饱和食盐水萃取有机相，无水硫酸钠干燥，硅胶柱纯化，得到产物APN。
反应瓶中依次加入化合物APN，2-溴乙醇和氯仿，80℃反应48h，减压除溶剂，制备硅胶层析柱纯化，得到产物APNO。尽管上面结合附图对本发明进行了描述，但是本发明并不局限于上述的具体实施方式，上述的具体实施方式仅仅是示意性的，而不是限制性的，本领域的普通技术人员在本发明的启示下，在不脱离本发明宗旨的情况下，还可以做出很多变形，这些均属于本发明的保护之内。
Claims (4)
1.一种新型AIE荧光探针分子，其结构式如下：
2.根据权利要求1所述的一种新型AIE荧光探针分子，其合成路线如下：
3.根据权利要求2所述的一种新型AIE荧光探针分子，其合成步骤如下：
1)对碘苯酚和碘代十六烷溶于DMF中，加入碳酸钾，室温反应过夜，后处理，得到化合物a；
2)化合物a和对溴苯胺溶于甲苯中，加入氢氧化钾，在1,10-菲咯啉和碘化亚铜催化下得到化合物b；
3)化合物b和5-醛基2-噻吩硼酸溶于甲醇和甲苯(v/v＝1:1)溶剂中，加入碳酸钾，在双三苯基膦二氯化钯催化下，60-90℃反应12-24h，后处理得到化合物c；
4)化合物c和对乙酸吡啶盐酸盐溶于氯仿中，加入哌啶为催化剂，60-80℃反应过夜，后处理得到化合物APN；
5)化合物APN溶于氯仿中，加入2-溴乙醇，60-80℃反应过夜，后处理得到AIE探针分子APNO。
4.根据权利要求1所述的一种新型AIE荧光探针分子的纳米胶束和囊泡的制备方法，其特征在于,包括如下步骤：
1)AIE探针分子APNO溶于四氢呋喃中，加入对应摩尔比的胆固醇:
其中，APNO/胆固醇的摩尔比范围为1:0-1:2；
2)磁力搅拌下，用注射器缓慢滴加去离子水，室温搅拌；
3)用去离子水透析，中间进行更换水，得到纳米胶束或囊泡。
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