CN105997888A - 一种巯基壳聚糖纳米-微球载体及其制备方法 - Google Patents

一种巯基壳聚糖纳米-微球载体及其制备方法 Download PDF

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CN105997888A
CN105997888A CN201610288148.9A CN201610288148A CN105997888A CN 105997888 A CN105997888 A CN 105997888A CN 201610288148 A CN201610288148 A CN 201610288148A CN 105997888 A CN105997888 A CN 105997888A
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王立强
李谦
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Abstract

本发明属于口服基因给药系统领域，具体涉及巯基壳聚糖纳米‑微球载体及其制备方法。载体是具有核壳结构的纳米‑微球，微球外壳为生物相容性好、可降解的聚己内酯(PCL)，内核为巯基壳聚糖纳米粒。核壳结构的纳米微球能够实现对药物的密封性保护，尤其适合胃肠道苛刻的生理环境，应用于基因口服载体等领域。

Description

一种巯基壳聚糖纳米-微球载体及其制备方法
技术领域
本发明属于口服基因给药系统领域，具体涉及一种巯基壳聚糖纳米-微球载体及其制备方法。
背景技术
十几年来，功能性载体材料不断涌现，但符合临床要求的基因口服制剂的研究仍举步维艰，主要原因包括：口服吸收机理不明确、给药系统的设计无所适从以及吸收、转染效率低下。基因药物与一般的小分子不同，导致其口服吸收的机理特殊并且复杂，还会受到药物性质、载体、配体以及个体差异等影响。胃肠道的生理学屏障是基因药物口服吸收的第一道屏障，胃肠道中的pH环境、各种酶都会对基因药物造成破坏。
发明内容
本发明的目的是提供一种在胃肠道内可有效封闭基因药物且不易降解，控释效果较好的巯基壳聚糖纳米-微球载体及其制备方法。
为了实现上述目的，本发明采用如下技术方案：
巯基壳聚糖纳米-微球载体，粒径为20-200μm，具有核壳结构，外壳为PCL，内核为巯基壳聚糖。
所述的PCL分子量为10000-50000，所述的巯基壳聚糖巯基取代度为30％-60％。
巯基壳聚糖纳米-微球载体的制备方法，包括如下步骤：
(1)将巯基壳聚糖与多聚磷酸钠在室温下，以500-2000rpm的转速搅拌15-30min后，经过透析、浓缩、冻干后得到巯基壳聚糖纳米粒；
(2)将上述巯基壳聚糖纳米粒通过去离子水复溶处理后，溶于质量体积比为0.5-10％的PCL二氯甲烷溶液，以2000-9000rpm的转速，20-25℃均化3-5min，形成稳定的乳化体系；
(3)再在上述稳定的乳化体系中加入质量体积比为0.1-1％的PVA水溶液，以2000-9000rpm的转速，20-25℃均化5min，然后以500-2000rpm的转速室温搅拌，直至二氯甲烷挥发完全，通过洗涤、离心制备得到巯基壳聚糖纳米-微球载体，此巯基壳聚糖纳米-微球载体粒径为20-200μm，具有核壳结构，外壳为PCL，内核为巯基壳聚糖。
所述的PCL分子量为10000-50000；所述的PVA醇解80-95％，分子量为20000-60000；所述的巯基壳聚糖巯基取代度为30％-60％。
微球的粒径大小根据所述PCL的质量、所述巯基壳聚糖纳米粒的质量以及所述步骤(2)与(3)中的转速进行控制，微球的成球性及表面光滑度根据所述PVA的质量分数控制。
采用上述方案后，本发明的巯基壳聚糖纳米-微球载体，外壳是PCL，PCL在胃内酸性pH中不溶，在肠道pH和脂肪酶的作用下才缓慢水解、酶解，加之PCL的疏水性，微球能为纳米微球在体内提供长时间的密闭保护，随着PCL在胃肠道中缓慢降解而释放出纳米粒，具有较好的控释效果，有利于基因长期表达，延长基因药物在体内的滞留时间，提高对DNA、载体的保护性，保证基因纳米载体在被细胞摄取前的完整性。核-壳结构本身有利于基因药物在体内的长期表达，并且该给药系统的制备由两步独立过程组成，先制备载药纳米粒，再以相对简单、惰性的条件包裹纳米粒形成微球，极大降低了制备工艺对基因药物的破坏，将反应路线中不可控的风险降至最低。不仅保留原载体巯基壳聚糖纳米粒的优点，还有助于跨越基因口服过程中的诸多屏障。
附图说明
图1-1为本发明中FITC标记的巯基壳聚糖纳米-微球载体的荧光视野下显微照片；图1-2为本品发明中FITC标记的巯基壳聚糖纳米-微球载体白光视野下显微照片。
图2-1为本发明中局部破碎的巯基壳聚糖纳米-微球载体的扫描电镜照片；图2-2为图2-1的局部放大示意图。
图3为本发明实施例1中的巯基壳聚糖纳米-微球载体的扫描电镜照片。
图4为本发明实施例2中的巯基壳聚糖纳米-微球载体的扫描电镜照片。
图5为本发明实施例3中的巯基壳聚糖纳米-微球载体的扫描电镜照片。
具体实施方式
为了更好地理解本发明，下面结合实例进一步阐明发明的内容，但是本发明的内容不仅局限于下面的实例：
实施例1：
巯基壳聚糖纳米-微球载体的制备方法，通过如下步骤实现：
(1)取0.1g巯基壳聚糖，巯基取代度30％，溶于0.1M乙酸，用2M氢氧化钠溶液调节溶液pH至5.0，与12mL1mg·mL-1多聚磷酸钠在室温下以2000rpm的转速搅拌20min，经过透析、浓缩、冻干后得到巯基壳聚糖纳米粒；
(2)取上述制备的巯基壳聚糖纳米粒0.1g通过500μL去离子水复溶处理后，溶于20mL，0.5％(W/V)PCL(分子量10000-20000)二氯甲烷溶液，以9000rpm的转速，20℃均化3min，形成稳定的乳化体系；
(3)再在上述稳定的乳化体系中加入20mL 0.1％(W/V)的PVA(醇解80％，分子量30000)水溶液，以9000rpm的转速，20℃均化5min，然后以2000rpm的转速室温搅拌，直至二氯甲烷挥发完全，通过洗涤、离心制备得到粒径约为20μm的巯基壳聚糖纳米-微球载体，其具有核壳结构，外壳为PCL，内核为巯基壳聚糖，扫描电镜照片如图3所示。
实施例2：
巯基壳聚糖纳米-微球载体的制备方法，通过如下步骤实现：
(1)取0.3g巯基壳聚糖(巯基取代度50％)，溶于0.1M乙酸，用2M氢氧化钠溶液调节溶液pH至5.0，与36mL1mg·mL-1多聚磷酸钠在室温下以2000rpm的转速搅拌20min，经过透析、浓缩、冻干后得到巯基壳聚糖纳米粒；
(2)取上述制备的巯基壳聚糖纳米粒0.3g通过500μL去离子水复溶处理后，溶于20mL，5％(W/V)PCL(分子量20000-40000)二氯甲烷溶液，以5000rpm的转速，20℃均化3min，形成稳定的乳化体系；
(3)再在上述稳定的乳化体系中加入20mL 0.1％(W/V)的PVA(醇解88％，分子量470000)水溶液，以5000rpm的转速，20℃均化5min，然后以1000rpm的转速室温搅拌，直至二氯甲烷挥发完全，通过洗涤、离心制备得到粒径约为100μm的巯基壳聚糖纳米-微球载体，其具有核壳结构，外壳为PCL，内核为巯基壳聚糖，扫描电镜照片如图4所示。
实施例3：
巯基壳聚糖纳米-微球载体的制备方法，通过如下步骤实现：
(1)取1.2g巯基壳聚糖(巯基取代度60％)，溶于0.1M乙酸，用2M氢氧化钠溶液调节溶液pH至5.0，与72mL1mg·mL-1多聚磷酸钠在室温下以2000rpm搅拌20min，经过透析、浓缩、冻干后巯基壳聚糖纳米粒；
(2)取上述制备的巯基壳聚糖纳米粒0.6g通过1mL去离子水复溶处理后，溶于20mL，10％(W/V)PCL(分子量40000-50000)二氯甲烷溶液，以3000rpm的转速，20℃均化5min，形成稳定的乳化体系；
(3)再在上述稳定的乳化体系中加入20mL 0.1％(W/V)的PVA(醇解88％，分子量470000)水溶液，以3000rpm的转速，20℃均化5min，然后以500rpm的转速室温搅拌，直至二氯甲烷挥发完全，通过洗涤、离心制备得到粒径约为200μm的巯基壳聚糖纳米-微球载体，其具有核壳结构，外壳为PCL，内核为巯基壳聚糖，扫描电镜照片如图5所示。
本发明中FITC标记的巯基壳聚糖纳米-微球载体的荧光视野下显微照片如图1-1所示，FITC标记的巯基壳聚糖纳米-微球载体白光视野下显微照片如图1-2所示；图2-1为本发明中局部破碎的巯基壳聚糖纳米-微球载体的扫描电镜照片；图2-2为图2-1的局部放大示意图。

Claims (5)

1.巯基壳聚糖纳米-微球载体，其特征在于：粒径为20-200μm，具有核壳结构，外壳为PCL，内核为巯基壳聚糖。
2.如权利要求1所述的巯基壳聚糖纳米-微球载体，其特征在于：所述的PCL分子量为10000-50000，所述的巯基壳聚糖巯基取代度为30％-60％。
3.巯基壳聚糖纳米-微球载体的制备方法，其特征在于，包括如下步骤：
(1)将巯基壳聚糖与多聚磷酸钠在室温下，以500-2000rpm的转速搅拌15-30min，制备得到巯基壳聚糖纳米粒；
(2)将上述巯基壳聚糖纳米粒依次经过透析、浓缩、冻干，再通过去离子水复溶处理后，溶于质量体积比为0.5-10％的PCL二氯甲烷溶液，以2000-9000rpm的转速，20-25℃均化3-5min，形成稳定的乳化体系；
(3)再在上述稳定的乳化体系中加入质量体积比为0.1-1％的PVA水溶液，以2000-9000rpm的转速，20-25℃均化5min，然后以500-2000rpm的转速室温搅拌，直至二氯甲烷挥发完全，通过洗涤、离心制备得到巯基壳聚糖纳米-微球载体，此巯基壳聚糖纳米-微球载体的粒径为20-200μm，具有核壳结构，外壳为PCL，内核为巯基壳聚糖。
4.如权利要求3所述的巯基壳聚糖纳米-微球载体的制备方法，所述的PCL分子量为10000-50000；所述的PVA醇解80-95％，分子量为20000-60000；所述的巯基壳聚糖巯基取代度为30％-60％。
5.如权利要求3所述的巯基壳聚糖纳米-微球载体的制备方法，其特征在于：微球的粒径大小根据所述PCL的质量、所述巯基壳聚糖纳米粒的质量以及所述步骤(2)与(3)中的转速进行控制，微球的成球性及表面光滑度根据所述PVA的质量分数控制。
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CN110318259A (zh) * 2019-06-25 2019-10-11 北京宇阳泽丽防水材料有限责任公司 一种高聚物改性沥青防水卷材及其生产工艺
CN110318259B (zh) * 2019-06-25 2021-12-07 北京宇阳泽丽防水材料有限责任公司 一种高聚物改性沥青防水卷材及其生产工艺

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