CN114366816A - 一种具有优良冻干再分散性的壳聚糖纳米粒、冻干制剂及其制备方法和应用 - Google Patents

一种具有优良冻干再分散性的壳聚糖纳米粒、冻干制剂及其制备方法和应用 Download PDF

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CN114366816A
CN114366816A CN202210009248.9A CN202210009248A CN114366816A CN 114366816 A CN114366816 A CN 114366816A CN 202210009248 A CN202210009248 A CN 202210009248A CN 114366816 A CN114366816 A CN 114366816A
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周兴
娄杰
甘峰旭
刘燕
邵文陶
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Chongqing Fumei Stem Cell Biotechnology Development Co ltd
Chongqing University of Technology
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Abstract

本发明提供了一种具有优良冻干再分散性的壳聚糖纳米粒、冻干制剂及其制备方法和应用，涉及壳聚糖纳米粒工艺技术领域。一种具有优良冻干再分散性的壳聚糖纳米粒的制备方法，其将亲水性壳聚糖和壳聚糖按比例混合，与三聚磷酸钠溶液按比例进行交联，制得壳聚糖纳米粒。本发明还提供了该具有优良冻干再分散性的壳聚糖纳米粒和其制得的冻干制剂及应用。本发明可以制备得到具有优良冻干再分散性的壳聚糖纳米粒，且在冻干时不使用冻干保护剂依然可以保持壳聚糖纳米粒原有的纳米结构。

Description

一种具有优良冻干再分散性的壳聚糖纳米粒、冻干制剂及其 制备方法和应用
技术领域
本发明涉及壳聚糖纳米粒工艺技术领域，具体而言，涉及一种具有优良冻干再分散性的壳聚糖纳米粒、冻干制剂及其制备方法和应用。
背景技术
离子凝胶法合成壳聚糖纳米粒反应条件温和、制备简单、无需使用有机溶剂，且由于其得到的纳米粒稳定性好、粒径均匀等优势成为最常用的制备方法，特别适合包载蛋白质、核酸、疫苗等大分子生化药物。蛋白因子主要成分为具有各种功能的多肽如PDGF和TGF-β等各种细胞因子。因此用离子凝胶法来合成负载蛋白因子的壳聚糖纳米粒是不二选择。壳聚糖具有良好的透膜能力可以有效地提高蛋白因子的透膜能力，进而提高蛋白因子在应用时的效果。
目前，用于制备壳聚糖纳米粒的壳聚糖为疏水性，其表面氨基之间的氢键作用过强，在冻干后纳米粒之间相互通过氢键交联，形成表面完全疏水的团聚体，导致壳聚糖纳米粒冻干后无法再分散于水中，冻干再分散性极差，限制了壳聚糖纳米粒的工业应用；另外，壳聚糖纳米粒在冻干时需要添加冻干保护剂才能保持壳聚糖纳米粒原有的纳米结构。
发明内容
本发明的第一个目的在于提供一种具有优良冻干再分散性的壳聚糖纳米粒的制备方法，可以制备得到具有优良冻干再分散性的壳聚糖纳米粒，且在冻干时不使用冻干保护剂依然可以保持壳聚糖纳米粒原有的纳米结构。
本发明的第二个目的在于提供一种具有优良冻干再分散性的壳聚糖纳米粒，通过制备方法制得的壳聚糖纳米粒，具有优良的冻干再分散性。
本发明的第三个目的在于提供一种壳聚糖纳米粒冻干制剂，其由壳聚糖纳米粒冻干制得。
本发明的第四个目的在于提供一种壳聚糖纳米粒冻干制剂在制备生化药物或护肤品中的应用。
本发明的实施例通过以下技术方案实现：
一种具有优良冻干再分散性的壳聚糖纳米粒的制备方法，包括以下步骤：
S1：将亲水性壳聚糖和壳聚糖按比例混合形成一定浓度的壳聚糖溶液；
S2：制备一定浓度的三聚磷酸钠溶液；
S3：按一定比例将三聚磷酸钠溶液缓慢滴加到壳聚糖溶液进行交联，制得壳聚糖纳米粒。
进一步地，所述步骤S1中亲水性壳聚糖为乳酸壳聚糖或羧甲基壳聚糖。
进一步地，所述步骤S1中亲水性壳聚糖和壳聚糖的质量比例为5～9:1～5。
进一步地，所述步骤S1制得的壳聚糖溶液的浓度为1～3mg/ml。
进一步地，所述步骤S2中制得的三聚磷酸钠溶液的浓度为1～5mg/ml。
进一步地，所述步骤S3中三聚磷酸钠溶液和壳聚糖溶液的体积比例为1:4～8。
进一步地，所述步骤S2中还包括将亲水药物或蛋白因子溶解于三聚磷酸钠溶液中制得载药三聚磷酸钠溶液，以使步骤S3制得载药壳聚糖纳米粒。
一种由制备方法制备得到的具有优良冻干再分散性的壳聚糖纳米粒或载药壳聚糖纳米粒。
一种壳聚糖纳米粒冻干制剂，由壳聚糖纳米粒冻干制得。
一种壳聚糖纳米粒冻干制剂在制备生化药物或护肤品中的应用。
本发明实施例的技术方案至少具有如下优点和有益效果：
本发明通过亲水性壳聚糖和壳聚糖按比例组装形成壳聚糖纳米粒，制备的壳聚糖纳米粒表面亲水，避免了壳聚糖纳米粒表面氢键的交联，减少纳米粒冻干后的交联现象，大幅提高了壳聚糖纳米粒的冻干再分散性能；且在冻干时不需要使用冻干保护剂，仍然可以保持壳聚糖纳米粒原有的纳米结构。
具体实施方式
为使本发明实施例的目的、技术方案和优点更加清楚，下面将对本发明实施例中的技术方案进行清楚、完整地描述。实施例中未注明具体条件者，按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者，均为可以通过市售购买获得的常规产品。
下面对本发明实施例提供的一种具有优良冻干再分散性的壳聚糖纳米粒、冻干制剂及其制备方法和应用进行具体说明。
实施例1
本实施例提供了一种具有优良冻干再分散性的壳聚糖纳米粒的制备方法，包括以下步骤：
S1：将乳酸壳聚糖和壳聚糖按质量比例为5:5混合形成1mg/ml浓度的壳聚糖溶液；
S2：制备1mg/ml浓度的三聚磷酸钠溶液；
S3：按体积比例为1:6将三聚磷酸钠溶液缓慢滴加到壳聚糖溶液进行交联，制得壳聚糖纳米粒A。
再将制得的壳聚糖纳米粒冻干制得壳聚糖纳米粒冻干制剂A1。
实施例2
本实施例提供了一种具有优良冻干再分散性的壳聚糖纳米粒的制备方法，包括以下步骤：
S1：将乳酸壳聚糖和壳聚糖按质量比例为7:3混合形成1mg/ml浓度的壳聚糖溶液；
S2：制备1mg/ml浓度的三聚磷酸钠溶液；
S3：按体积比例为1:6将三聚磷酸钠溶液缓慢滴加到壳聚糖溶液进行交联，制得壳聚糖纳米粒B。
再将制得的壳聚糖纳米粒冻干制得壳聚糖纳米粒冻干制剂B1。
实施例3
本实施例提供了一种具有优良冻干再分散性的壳聚糖纳米粒的制备方法，包括以下步骤：
S1：将乳酸壳聚糖和壳聚糖按质量比例为7:3混合形成1mg/ml浓度的壳聚糖溶液；
S2：制备1mg/ml浓度的三聚磷酸钠溶液，再于三聚磷酸钠溶液中溶解蛋白因子至终浓度1mg/ml，即制得载药三聚磷酸钠溶液；
S3：按体积比例为1:6将载药三聚磷酸钠溶液缓慢滴加到壳聚糖溶液进行交联，制得载药壳聚糖纳米粒C。
再将制得的壳聚糖纳米粒冻干制得壳聚糖纳米粒冻干制剂C1。
实施例4
本实施例提供了一种具有优良冻干再分散性的壳聚糖纳米粒的制备方法，包括以下步骤：
S1：将乳酸壳聚糖和壳聚糖按质量比例为9:1混合形成1mg/ml浓度的壳聚糖溶液；
S2：制备1mg/ml浓度的三聚磷酸钠溶液，再于三聚磷酸钠溶液中溶解蛋白因子至终浓度1mg/ml，即制得载药三聚磷酸钠溶液；
S3：按体积比例为1:6将载药三聚磷酸钠溶液缓慢滴加到壳聚糖溶液进行交联，制得载药壳聚糖纳米粒D。
再将制得的壳聚糖纳米粒冻干制得壳聚糖纳米粒冻干制剂D1。
对比例1
本对比例提供了一种壳聚糖纳米粒的制备方法，包括以下步骤：
S1：将壳聚糖溶解于1％醋酸溶剂中形成1mg/ml浓度的壳聚糖溶液；
S2：制备1mg/ml浓度的三聚磷酸钠溶液，再于三聚磷酸钠溶液中溶解蛋白因子至终浓度1mg/ml，即制得载药三聚磷酸钠溶液；
S4：按体积比例为1:6将载药三聚磷酸钠溶液缓慢滴加到壳聚糖溶液进行交联，制得载药壳聚糖纳米粒M；将载药壳聚糖纳米粒M中按质量比10％混合加入甘露醇。
再将混合加入甘露醇的壳聚糖纳米粒冻干制得壳聚糖纳米粒冻干制剂M1。
对比例2
本对比例提供了一种壳聚糖纳米粒的制备方法，包括以下步骤：
S1：将壳聚糖溶解于1％醋酸溶剂中形成1mg/ml浓度的壳聚糖溶液；
S2：制备1mg/ml浓度的三聚磷酸钠溶液，再于三聚磷酸钠溶液中溶解蛋白因子至终浓度1mg/ml，即制得载药三聚磷酸钠溶液；
S3：按体积比例为1:6将载药三聚磷酸钠溶液缓慢滴加到壳聚糖溶液进行交联，制得载药壳聚糖纳米粒G。
再将制得的壳聚糖纳米粒冻干制得壳聚糖纳米粒冻干制剂G1。
实验例1
取等量的实施例1～4制得的壳聚糖纳米粒A～D和对比例1～2制得的壳聚糖纳米粒M、G，检测各壳聚糖纳米粒的多分散指数；再将等量的实施例1～4制得的壳聚糖纳米粒冻干制剂A1～D1和对比例1～2制得的壳聚糖纳米粒冻干制剂M1、G1分别用水溶液复溶，检测各壳聚糖纳米粒冻干制剂复溶溶液的多分散指数；结果如表1所示。
表1 多分散指数
Figure BDA0003456714360000071
Figure BDA0003456714360000081
由表1可知，本发明实施例1～4制得的壳聚糖纳米粒A～D的粒径和实施例1～4制得的壳聚糖纳米粒冻干制剂A1～D1复溶溶液的多分散指数变化不大，且均较小，在0.4以下，而对比例1～2制得的壳聚糖纳米粒冻干制剂M1、G1复溶溶液的粒径比对比例1～2制得的壳聚糖纳米粒M、G的多分散指数增大较多，且超过0.6；说明本发明方法制得的壳聚糖纳米粒冻干制剂复溶后粒径分布范围仍然较集中，说明本发明方法制得的壳聚糖纳米粒的冻干再分散性能较优良。
实验例2
取等量的实施例1～4制得的壳聚糖纳米粒A～D和对比例1～2制得的壳聚糖纳米粒M、G，检测各壳聚糖纳米粒的粒径；再将等量的实施例1～4制得的壳聚糖纳米粒冻干制剂A1～D1和对比例1～2制得的壳聚糖纳米粒冻干制剂M1、G1分别用水溶液复溶，检测各壳聚糖纳米粒冻干制剂复溶溶液的粒径；结果如表2所示。
表2 粒径
Figure BDA0003456714360000091
由表2可知，本发明实施例1～4制得的壳聚糖纳米粒A～D的粒径和实施例1～4制得的壳聚糖纳米粒冻干制剂A1～D1复溶溶液的粒径变化不大，对比例1制得的壳聚糖纳米粒M在冻干时采用了甘露醇冻干保护剂，其制得的壳聚糖纳米粒冻干制剂M1复溶溶液的粒径变化不大，而对比例2制得的壳聚糖纳米粒G在冻干时未采用冻干保护剂，其制得的壳聚糖纳米粒冻干制剂G1复溶溶液的粒径比对比例2制得的壳聚糖纳米粒G的粒径增大较多；说明本发明方法在冻干时不使用冻干保护剂制得的壳聚糖纳米粒仍然可以保持壳聚糖纳米粒原有的纳米结构。
综上，通过亲水性壳聚糖和壳聚糖按比例组装形成壳聚糖纳米粒，制备的壳聚糖纳米粒具有优良的冻干再分散性能，且在冻干时不需要使用冻干保护剂，仍然可以保持壳聚糖纳米粒原有的纳米结构。
以上所述仅为本发明的优选实施例而已，并不用于限制本发明，对于本领域的技术人员来说，本发明可以有各种更改和变化。凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。

Claims (10)

1.一种具有优良冻干再分散性的壳聚糖纳米粒的制备方法，其特征在于，包括以下步骤：
S1：将亲水性壳聚糖和壳聚糖按比例混合形成一定浓度的壳聚糖溶液；
S2：制备一定浓度的三聚磷酸钠溶液；
S3：按一定比例将三聚磷酸钠溶液缓慢滴加到壳聚糖溶液进行交联，制得壳聚糖纳米粒。
2.根据权利要求1所述的具有优良冻干再分散性的壳聚糖纳米粒的制备方法，其特征在于，所述步骤S1中亲水性壳聚糖为乳酸壳聚糖或羧甲基壳聚糖。
3.根据权利要求1所述的具有优良冻干再分散性的壳聚糖纳米粒的制备方法，其特征在于，所述步骤S1中亲水性壳聚糖和壳聚糖的质量比例为5～9:1～5。
4.根据权利要求1所述的具有优良冻干再分散性的壳聚糖纳米粒的制备方法，其特征在于，所述步骤S1制得的壳聚糖溶液的浓度为1～3mg/ml。
5.根据权利要求1所述的具有优良冻干再分散性的壳聚糖纳米粒的制备方法，其特征在于，所述步骤S2中制得的三聚磷酸钠溶液的浓度为1～5mg/ml。
6.根据权利要求1所述的具有优良冻干再分散性的壳聚糖纳米粒的制备方法，其特征在于，所述步骤S3中三聚磷酸钠溶液和壳聚糖溶液的体积比例为1:4～8。
7.根据权利要求1所述的具有优良冻干再分散性的壳聚糖纳米粒的制备方法，其特征在于，所述步骤S2中还包括将亲水药物或蛋白因子溶解于三聚磷酸钠溶液中制得载药三聚磷酸钠溶液，以使步骤S3制得载药壳聚糖纳米粒。
8.一种由权利要求1～7任一项所述的制备方法制备得到的具有优良冻干再分散性的壳聚糖纳米粒或载药壳聚糖纳米粒。
9.一种壳聚糖纳米粒冻干制剂，其特征在于，由权利要求1～7任一项所述的制备方法制得的壳聚糖纳米粒或载药壳聚糖纳米粒或权利要求8所述的壳聚糖纳米粒或载药壳聚糖纳米粒冻干制得。
10.一种权利要求9所述的壳聚糖纳米粒冻干制剂在制备生化药物或护肤品中的应用。
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