CN113855706A - 一种具有化学动力学治疗及化疗作用纳米粒的制备方法 - Google Patents
一种具有化学动力学治疗及化疗作用纳米粒的制备方法 Download PDFInfo
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Abstract
本发明涉及纳米药物技术领域，具体涉及一种具有化学动力学治疗及化疗作用纳米粒的制备方法。包括以下几个步骤（1）黑色素药物载体的合成；（2）载体的纯化；(3)纳米粒的自组装；(4)纳米粒的纯化；(5)纳米粒的冷冻干燥。本发明通过合成具有肿瘤化学动力学治疗作用的水溶天然黑色素药物载体，并通过纳米共沉淀法使之与化疗药物自组装为可联合化学动力疗法与化疗的纳米粒子，本发明可为多功能肿瘤治疗平台的设计提供参考。
Description
技术领域
本发明涉及纳米药物技术领域，具体涉及一种具有化学动力学治疗及化疗作用纳米粒的制备方法。
背景技术
化疗是恶性肿瘤治疗的主要策略，然而，化疗药较大的副作用、较低的生物利用度限制了其疗效。纳米药物递送***因可同时改善药物的肿瘤靶向性、控释性、生物利用度、溶解性等而备受青睐。此外，纳米药物递送***可实现化疗与肿瘤的其他新兴疗法如光热治疗、光动力治疗、声动力治疗、化学动力学治疗的联用，从而避免单一疗法引发的肿瘤多重耐药。化学动力学疗法指的是利用纳米催化剂将肿瘤部位过量的过氧化氢转化为高毒性的羟自由基，进而对肿瘤产生杀灭作用。该疗法无需外源能量的输入、毒性低、肿瘤选择性高，较适合作为化疗的联合疗法。蜣螂黑色素是一种富含铁离子且对过氧化氢敏感的色素，具有优良的芬顿反应催化作用且可通过疏水作用、π-π堆积力、氢键等非共价键与多种化合物产生作用。然而，蜣螂黑色素的水溶性较差，具有一定的局限性。为此，本发明将采用化学修饰方法提高蜣螂黑色素的亲水性，并将其与抗肿瘤药物阿霉素自组装为可联合化疗与化学动力学疗法的抗肿瘤纳米粒，本发明将为肿瘤化疗药的纳米递送提供参考。
发明内容
本发明提供一种具有化学动力学治疗及化疗作用纳米粒的制备方法，可获得联合化疗与化学动力学疗法的抗肿瘤纳米粒，将为多功能肿瘤治疗平台的设计及昆虫黑色素的药用提供参考。
一种具有化学动力学治疗及化疗作用纳米粒的制备方法，包括以下步骤：
（1）黑色素药物载体的合成：将蜣螂黑色素按料液比1/100～2/25加入pH 8～9的氨水中，超声30～60分钟，超声功率300～500W，抽滤，得黑色素溶液；将分子量为1000～2000 Da的壳寡糖按料液比1/20～2/5溶于去离子水中，得壳寡糖溶液；取黑色素溶液，按体积比1/3～1/1缓慢加入壳寡糖溶液，同时持续搅拌12～24小时，得黑色素的壳寡糖修饰物溶液。
（2）载体的纯化：将（1）中的黑色素的壳寡糖修饰物溶液，置于截留分子量3500～7000Da的透析袋中，流动去离子水透析48～72小时，随后，取透析袋内容物，冷冻干燥，得壳寡糖修饰的黑色素载体。
（3）纳米粒的自组装：将冻干的壳寡糖修饰的黑色素载体按料液比1/100～1/20分散于50％～75％的二甲基亚砜水溶液（V：V），持续搅拌30～80分钟，得黑色素载体溶液。将阿霉素按料液比1/50～1/10溶于二甲基亚砜中，得阿霉素溶液。随后，取黑色素载体溶液，按体积比3/1～5/1缓慢加入阿霉素溶液，同时持续搅拌20～60分钟，接下来，将黑色素载体与阿霉素混合液置于截留分子量3500～10000 Da的透析袋，流动的去离子水透析48～72小时，透析袋内容物即为纳米粒粗品。
（4）纳米粒的纯化：将纳米粒粗品置于截留分子量为10～100 Kda的超滤离心管中，3000转/分钟～5000转/分钟的转速下离心20～45分钟，弃去滤液，即得精制的纳米粒悬液。
（5）纳米粒的冷冻干燥：往精制的纳米粒悬液中按料液比1/100～3/100加入甘露糖，并进行冷冻干燥，冷阱温度-70℃～50℃。
具体实施方式
下面结合具体实施方式对本发明进行详细说明。
实施例一
（1）黑色素药物载体的合成：将蜣螂黑色素按料液比1/100加入pH 8的氨水中，超声30分钟，超声功率300W，抽滤，得黑色素溶液；将分子量为1000 Da的壳寡糖按料液比1/20溶于去离子水中，得壳寡糖溶液；取黑色素溶液，按体积比1/3缓慢加入壳寡糖溶液，同时持续搅拌12小时，得黑色素的壳寡糖修饰物溶液。
（2）载体的纯化：将（1）中的黑色素的壳寡糖修饰物溶液，置于截留分子量3500Da的透析袋中，流动去离子水透析48小时，随后，取透析袋内容物，冷冻干燥，得壳寡糖修饰的黑色素载体。
（3）纳米粒的自组装：将冻干的壳寡糖修饰的黑色素载体按料液比1/100分散于50％的二甲基亚砜水溶液（V：V），持续搅拌30分钟，得黑色素载体溶液。将阿霉素按料液比1/50溶于二甲基亚砜中，得阿霉素溶液。随后，取黑色素载体溶液，按体积比3/1缓慢加入阿霉素溶液，同时持续搅拌20分钟，接下来，将黑色素载体与阿霉素混合液置于截留分子量3500Da的透析袋，流动的去离子水透析48小时，透析袋内容物即为纳米粒粗品。
（4）纳米粒的纯化：将纳米粒粗品置于截留分子量为10Kda的超滤离心管中，3000转/分钟的转速下离心20分钟，弃去滤液，即得精制的纳米粒悬液。
（5）纳米粒的冷冻干燥：往精制的纳米粒悬液中按料液比1/100加入甘露糖、甘露醇，并进行冷冻干燥，冷阱温度-70℃。
实施例二
（1）黑色素药物载体的合成：将蜣螂黑色素按料液比1/20加入pH 8.5的氨水中，超声45分钟，超声功率450W，抽滤，得黑色素溶液；将分子量为2000 Da的壳寡糖按料液比1/4溶于去离子水中，得壳寡糖溶液；取黑色素溶液，按体积比1/2缓慢加入壳寡糖溶液，同时持续搅拌20小时，得黑色素的壳寡糖修饰物溶液。
（2）载体的纯化：将（1）中的黑色素的壳寡糖修饰物溶液，置于截留分子量5000Da的透析袋中，流动去离子水透析60小时，随后，取透析袋内容物，冷冻干燥，得壳寡糖修饰的黑色素载体。
（3）纳米粒的自组装：将冻干的壳寡糖修饰的黑色素载体按料液比2/50分散于60％的二甲基亚砜水溶液（V：V），持续搅拌60分钟，得黑色素载体溶液。将阿霉素按料液比2/25溶于二甲基亚砜中，得阿霉素溶液。随后，取黑色素载体溶液，按体积比4/1缓慢加入阿霉素溶液，同时持续搅拌45分钟，接下来，将黑色素载体与阿霉素混合液置于截留分子量7000Da的透析袋，流动的去离子水透析60小时，透析袋内容物即为纳米粒粗品。
（4）纳米粒的纯化：将纳米粒粗品置于截留分子量为100 Kda的超滤离心管中，4000转/分钟的转速下离心30分钟，弃去滤液，即得精制的纳米粒悬液。
（5）纳米粒的冷冻干燥：往精制的纳米粒悬液中按料液比1/50加入甘露醇，并进行冷冻干燥，冷阱温度-60℃。
本发明通过化学修饰法制备了水溶的壳寡糖修饰的黑色素载体，并采用纳米共沉淀法制备了载有阿霉素，兼有化疗与化学动力学治疗作用的黑色素纳米粒子，将为多功能抗癌纳米粒子的构建提供参考。
本发明不局限于上述具体的实施方式，本发明可以有各种更改和变化。凡是依据本发明的技术实质对以上实施方式所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围。
Claims (6)
1.一种具有化学动力学治疗及化疗作用纳米粒的制备方法，包括以下步骤：
黑色素药物载体的合成：制备了壳寡糖修饰的黑色素药物载体；
载体的纯化：利用透析法除去游离的壳寡糖及黑色素；
纳米粒的自组装：纳米共沉淀法构建了黑色素与阿霉素的纳米粒；
纳米粒的纯化：利用超滤法除去纳米粒中游离的阿霉素与黑色素载体；
纳米粒的冷冻干燥：加入甘露糖或甘露醇抗冻剂，并冻干纳米粒。
2.根据权利1所述一种具有化学动力学治疗及化疗作用纳米粒的制备方法，其特征在于，步骤（1）所述的黑色素药物载体的合成：将蜣螂黑色素按料液比1/100～2/25加入pH 8～9的氨水中，超声30～60分钟，超声功率300～500W，抽滤，得黑色素溶液；将分子量为1000～2000 Da的壳寡糖按料液比1/20～2/5溶于去离子水中，得壳寡糖溶液；取黑色素溶液，按体积比1/3～1/1缓慢加入壳寡糖溶液，同时持续搅拌12～24小时，得黑色素的壳寡糖修饰物溶液。
3.根据权利1所述一种具有化学动力学治疗及化疗作用纳米粒的制备方法，其特征在于，步骤（2）载体的纯化：将（1）中的黑色素的壳寡糖修饰物溶液，置于截留分子量3500～7000Da的透析袋中，流动去离子水透析48～72小时，随后，取透析袋内容物，冷冻干燥，得壳寡糖修饰的黑色素载体。
4.根据权利1所述一种具有化学动力学治疗及化疗作用纳米粒的制备方法，其特征在于，步骤（3）纳米粒的自组装：将冻干的壳寡糖修饰的黑色素载体按料液比1/100～1/20分散于50％～75％的二甲基亚砜水溶液V：V，持续搅拌30～80分钟，得黑色素载体溶液，将阿霉素按料液比1/50～1/10溶于二甲基亚砜中，得阿霉素溶液，随后，取黑色素载体溶液，按体积比3/1～5/1缓慢加入阿霉素溶液，同时持续搅拌20～60分钟，接下来，将黑色素载体与阿霉素混合液置于截留分子量3500～10000 Da的透析袋，流动的去离子水透析48～72小时，透析袋内容物即为纳米粒粗品。
5.根据权利1所述一种具有化学动力学治疗及化疗作用纳米粒的制备方法，其特征在于，步骤（4）纳米粒的纯化：将纳米粒粗品置于截留分子量为10～100 Kda的超滤离心管中，3000转/分钟～5000转/分钟的转速下离心20～45分钟，弃去滤液，即得精制的纳米粒悬液。
6.根据权利1所述一种具有化学动力学治疗及化疗作用纳米粒的制备方法，其特征在于，步骤（5）纳米粒的冷冻干燥：往精制的纳米粒悬液中按料液比1/100～3/100加入甘露糖，并进行冷冻干燥，冷阱温度-70℃～50℃。
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| CN107998392A (zh) * | 2017-12-08 | 2018-05-08 | 同济大学 | 具有增强光吸收的黑色素/Ce6光动力纳米药物及其制备方法 |
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| CN107998392A (zh) * | 2017-12-08 | 2018-05-08 | 同济大学 | 具有增强光吸收的黑色素/Ce6光动力纳米药物及其制备方法 |
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