CN113368056A - 一种负载巴诺蒽醌的无载体纳米递送系统及其制备方法 - Google Patents
一种负载巴诺蒽醌的无载体纳米递送系统及其制备方法 Download PDFInfo
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Abstract
本发明公开了一种负载巴诺蒽醌的无载体纳米递送系统及其制备方法,包括如下步骤:(1)取巴诺蒽醌·二盐酸盐加入水中,搅拌均匀;(2)向步骤(1)的溶液中加入二氯化锰溶液,搅拌进行反应,再加入三乙胺,搅拌反应,以去除HCl;(3)将步骤(2)所得的反应液缓慢滴加至水中,搅拌进行反应,最后透析除去未配位的巴诺蒽醌,得到所述负载巴诺蒽醌的纳米递送系统。本发明通过金属配位的方法直接将巴诺蒽醌与金属锰离子构成金属有机框架(MOFs),制得负载巴诺蒽醌的无载体纳米递送系统(AMMOFs),AMMOFs在肿瘤细胞内涵体微酸性环境中可有效释放药物,具有造影成像、治疗作用,载药量高,且合成工艺简单。
Description
技术领域
本发明涉及肿瘤治疗及纳米药物递送技术领域,特别是涉及一种负载巴诺蒽醌的无载体纳米递送系统及其制备方法。
背景技术
恶性肿瘤是威胁人类生命健康的头号杀手。缺氧是大多数实体肿瘤的显著特征,是肿瘤的发展和转移、治疗失败的重要原因。针对肿瘤缺氧开展治疗,已经成为研究肿瘤诊疗的新方向。目前最重要的一类以肿瘤缺氧为靶点的治疗方式为缺氧激活前药,如:巴诺蒽醌(Banox antrone,AQ4N),替拉扎明(Tirapazamine,TPZ)等,该类药物对缺氧细胞具有选择性和高毒性,而对富氧区域如正常组织和细胞却展示出低毒性,但单纯给药生物利用度低。纳米递送系统可有效提高其生物利用度,但目前所报道的纳米递送系统存在载药量低、合成工艺复杂的缺陷。
Jiong Li等人研究的含有聚乙二醇(PEG)和聚己内酯(PLC)的三嵌段共聚物纳米系统,可负载巴诺蒽醌(AQ4N)和葡萄糖氧化酶(GOD)[“Adv Healthc Mater”,(10):e1801627,(2019年)],但合成工艺复杂,且AQ4N载药量仅8.17%。Michael A.等人研究由巨噬细胞介导的缺氧激活前药纳米递送系统[“Advanced Therapeutics”,第3卷第2期:1900162,(2019年)]能增强TPZ在实体肿瘤中的渗透和聚集,其载药量为40.1%。
发明内容
鉴于以上所述现有技术的缺点,本发明的目的在于提供一种负载巴诺蒽醌的无载体纳米递送系统AMMOFs及其制备方法,所述纳米递送系统通过金属配位的方法直接将巴诺蒽醌与金属锰离子构成金属有机框架(Metal-Organic Frameworks,MOFs),载药量高,在肿瘤细胞内涵体微酸性环境中(pH=5.0)可有效释放药物,且合成工艺简单。
为实现上述目的及其他相关目的,本发明第一方面提供一种负载巴诺蒽醌的无载体纳米递送系统的制备方法,包括如下步骤:
(1)取巴诺蒽醌·二盐酸盐(AQ4N·HCl)加入水中,搅拌溶解;
(2)向步骤(1)的溶液中逐滴滴加二氯化锰溶液,搅拌进行反应,再逐滴滴加三乙胺,搅拌反应,以去除HCl;
(3)将步骤(2)所得的反应液缓慢滴加至水中,搅拌进行反应,最后透析除去未配位的巴诺蒽醌,得到所述负载巴诺蒽醌的纳米递送系统。
进一步,所述步骤(1)、(2)中,巴诺蒽醌·二盐酸盐与二氯化锰的质量比为20-50:1,优选为20:1。
进一步,所述步骤(1)中,所述巴诺蒽醌·二盐酸盐的浓度为2mg/mL。进一步,所述步骤(1)中,搅拌时间为1-2h,优选为1h。
进一步,所述步骤(2)中,二氯化锰溶液的浓度为1mg/mL。
进一步,所述步骤(2)中,巴诺蒽醌·二盐酸盐与三乙胺的用量比为1:(0.22-0.23)(mg/mL)。
进一步,所述步骤(2)中,二氯化锰溶液加入后,继续搅拌反应3-5h,优选为4h。
进一步,所述步骤(2)中,三乙胺加入后,继续搅拌反应不低于8h。
进一步,所述步骤(3)中,步骤(2)得到的反应液与水的体积用量比为1:4-6,优选为1:5。
进一步,所述步骤(3)中,搅拌反应时间为2-3h。
进一步,所述步骤(3)中,透析次数不低于3次,每次透析时间为不低于15min。
进一步,所述步骤(1)、(2)、(3)中,反应全程需要在避光条件下进行。
进一步,所述步骤(1)、(2)、(3)都在室温(25℃)下进行。
进一步,所述纳米递送系统的粒径为118±22.86纳米。
本发明第二方面提供一种根据第一方面所述的制备方法制得的负载巴诺蒽醌的无载体纳米递送系统。
如上所述,本发明的负载巴诺蒽醌的无载体纳米递送系统及其制备方法,具有以下有益效果:
1、本发明通过金属配位的方法直接将巴诺蒽醌与金属锰离子构成金属有机框架(Metal-Organic Frameworks,MOFs),制得一种新型的负载巴诺蒽醌的无载体化纳米载药系统AMMOFs,Mn2+由于金属阳离子(Mn2+)与药物形成的配位键具有酸敏感性,所以本发明的AMMOFs可在肿瘤组织(肿瘤细胞内涵体)弱酸性环境解体,有效释放药物;同时AMMOFs能为肿瘤组织靶向提供吸光物质AQ4N,以提高光声成像效果,因此,本发明的AMMOFs具有造影成像、治疗等作用。
2、本发明所述方法制备的无载体化纳米载药系统,载药量高,载药后形成的纳米粒子粒度均匀,直径小,分散好,不团聚,有利于提高治疗效果。
3、本发明的无载体化纳米载药系统制备方法简单,且原料易于获取,使用的设备为常规设备,有利于大规模工业化生产。
附图说明
图1显示为本发明实施例1中负载巴诺蒽醌的无载体纳米递送系统的粒径分布图。
图2显示为本发明实施例1中负载巴诺蒽醌的无载体纳米递送系统的扫描电镜图。
图3显示为本发明实施例1中细胞摄取激光共聚焦图。
图4显示为本发明实施例1中负载巴诺蒽醌的无载体纳米递送系统的体内光声成像图。
图5显示为本发明实施例中巴诺蒽醌的标准曲线图。
具体实施方式
以下结合附图和下述实施方式进一步说明本发明,应理解,本领域技术人员可由本说明书所揭露的内容轻易地了解本发明的其他优点与功效。本发明还可以通过另外不同的具体实施方式加以实施或应用,本说明书中的各项细节也可以基于不同观点与应用,在没有背离本发明的精神下进行各种修饰或改变。
本发明提供了一种负载巴诺蒽醌的无载体纳米递送系统,其制备方法包括如下步骤:
(1)取巴诺蒽醌·二盐酸盐(AQ4N·HCl)加入水中,搅拌以溶解充分;
(2)向步骤(1)的溶液中加入二氯化锰溶液,搅拌进行反应,再加入三乙胺,搅拌反应,以去除HCl;
(3)将步骤(2)所得的反应液缓慢滴加至水中,搅拌进行反应,最后透析除去未配位的巴诺蒽醌,得到所述负载巴诺蒽醌的纳米递送系统。
进一步,所述步骤(1)、(2)中,巴诺蒽醌·二盐酸盐与二氯化锰的质量比为20-50:1,优选为20-30:1,更优选为20:1。
进一步地,所述步骤(1)中,所述巴诺蒽醌的浓度为2mg/mL。
进一步地,所述步骤(1)中,搅拌时间为1-2h,优选为1h。
进一步地,所述步骤(2)中,二氯化锰溶液的浓度为1mg/mL。
进一步地,所述步骤(2)中,巴诺蒽醌·二盐酸盐与三乙胺的用量比为1:(0.22-0.23)(mg/mL)。
进一步地,所述步骤(2)中,二氯化锰溶液加入后,继续搅拌反应3-5h,优选为4h。
进一步地,所述步骤(2)中,三乙胺加入后,继续搅拌反应不低于8h。
进一步地,所述步骤(3)中,步骤(2)得到的反应液与水的体积用量比为1:4-6,优选为1:5。
进一步地,所述步骤(3)中,搅拌反应时间为2-3h。
进一步地,所述步骤(3)中,透析次数不低于3次,每次透析时间为不低于15min。
需要说明的是,本发明的制备方法中,所采用的水选自去离子水、超纯水;透析时使用的透析袋为MD44(3500D),来自于Solarbio;所涉及的反应及操作都在是室温(25℃)下进行的,反应全程在避光条件下进行。
本发明通过金属配位的方法直接将巴诺蒽醌与金属锰离子构成金属有机框架(Metal-Or ganic Frameworks,MOFs),制得一种新型的负载巴诺蒽醌的无载体化纳米载药系统,载药量高,可在肿瘤组织(肿瘤细胞内涵体)弱酸性环境解体,有效释放药物,同时具有造影成像、治疗作用。同时,本发明制备方法简单,制得的无载体化纳米载药系统,载药后形成的纳米粒子粒度均匀,直径小,分散好,不团聚,有利于提高治疗效果。
下面进一步地例举实施例以详细说明本发明。同样应理解,以下实施例只用于对本发明进行进一步地说明,不能理解为对本发明保护范围的限制,本领域的技术人员根据本发明的上述内容作出的一些非本质的改进和调整均属于本发明的保护范围。下述示例具体的工艺参数等也仅是合适范围中的一个示例,即本领域技术人员可以通过本文的说明做合适的范围内选择,而并非要限定于下文示例的具体数值。
实施例1
本实施例中负载巴诺蒽醌的无载体纳米递送系统的制备方法,包括如下步骤:
(1)取2mg巴诺蒽醌·二盐酸盐(AQ4N·HCl)于棕色玻璃瓶中,加入1mL去离子水溶解,并磁力搅拌。
(2)搅拌1h后,逐滴加入100μl的1mg/mL的二氯化锰溶液,搅拌4h后,逐滴加入450μL三乙胺,继续搅拌过夜,去除HCl。
(3)然后向盛有10mL去离子水的圆底烧瓶中,缓慢滴加2mL步骤(2)所得的反应液,进行磁力搅拌,搅拌时间为2h。
(4)最后将步骤(3)所得反应液用透析袋透析,以去除未配位的巴诺蒽醌,得到负载巴诺蒽醌的纳米递送系统。
上述过程中,反应需全程避光,搅拌时转速均控制在700-800rpm,具体为750rpm。
实施例2
本实施例中负载巴诺蒽醌的无载体纳米递送系统的制备方法,包括如下步骤:
(1)取2mg巴诺蒽醌·二盐酸盐(AQ4N·HCl)于棕色玻璃瓶中,加入1mL去离子水溶解,并磁力搅拌。
(2)搅拌2h后,逐滴加入100μL的1mg/mL的二氯化锰溶液,搅拌3h后,逐滴加入450μL三乙胺,继续搅拌过夜,去除HCl。
(3)然后向盛有10mL去离子水的圆底烧瓶中,缓慢滴加2mL步骤(2)所得的反应液,进行磁力搅拌,搅拌时间为2h。
(4)最后将步骤(3)所得反应液用透析袋透析,以去除未配位的巴诺蒽醌,得到负载巴诺蒽醌的纳米递送系统。
上述过程中,反应需全程避光,搅拌时转速均控制在700-800rpm,具体为750rpm。
实施例3
本实施例中负载巴诺蒽醌的无载体纳米递送系统的制备方法,包括如下步骤:
(1)取20mg巴诺蒽醌·二盐酸盐(AQ4N·HCl)于棕色玻璃瓶中,加入10mL去离子水溶解,并磁力搅拌。
(2)搅拌2h后,逐滴加入1mL的1mg/mL的二氯化锰溶液,搅拌5h后,逐滴加入4.5mL三乙胺,继续搅拌过夜,去除HCl。
(3)然后向盛有10mL去离子水的圆底烧瓶中,缓慢滴加2mL步骤(2)所得的反应液,进行磁力搅拌,搅拌时间为3h。
(4)最后将步骤(3)所得反应液用透析袋透析,以去除未配位的巴诺蒽醌,得到负载巴诺蒽醌的纳米递送系统。
上述过程中,反应需全程避光,搅拌时转速均控制在700-800rpm,具体为750rpm。
将实施例1制得的负载巴诺蒽醌的纳米递送系统(简称为AMMOFs)进行试验,实验过程、实验结果及分析如下:
1、用Marven激光粒度仪测定实施例1制得的AMMOFs的平均粒径、分布,结果如图1所示。
从图1的粒径分布图可知,负载巴诺蒽醌的无载体纳米递送系统的粒径大小在118±22.86纳米,粒径较小,分布较窄,有利于体内的靶向分布。
2、用TEM观察实施例1制得的AMMOFs的形貌特征,结果如图2所示。
从图2的电镜图可以看出,负载巴诺蒽醌的无载体纳米递送系统的粒径大小均一,呈球形,分散性良好。
3、细胞分布实验:
将2.0mL MDA-MB-231细胞(初始密度为104/孔)接种到共聚焦培养皿中。24h后将各皿培养基分别替换为free AQ4N和AMMOFs(AQ4N等量剂量:25ug/ml)的新鲜培养基溶液,再培养4h。随后移除培养液,用PBS轻洗细胞两次,加入1mL甲醇固定1min,PBS洗1次,随后加入200ul DAPI染色12min,再用PBS轻洗细胞两次,加PBS 1ml,使用激光共聚焦扫描成像(CLSM)观察荧光。
图3为激光共聚焦图,即药物在肿瘤细胞内的分布图,其中蓝色代表DAPI,红色代表AQ4N,紫色代表merge,是DAPI和AQ4N融合后的图像。从图3中可知,负载巴诺蒽醌的无载体纳米递送系统AMMOFs在与癌细胞作用4个小时后,药物明显进入细胞核,表示药物在细胞内进行了有效释放。
4、荷瘤鼠光声成像实验
选择荷瘤裸鼠进行实验,经尾静脉注射200μL AMMOFs(AQ4N等效剂量:25mg/L),然后在Vevo Laser光声成像系统上用690nm激光照射采集8h的光声成像(PA)图,如图4所示。
图4为荷瘤鼠光声成像图,其中红色代表具有光声成像效果,这表明本发明的ADMOPs具备光声成像效果,8h后ADMOPs到达了肿瘤区域。
5、AQ4N标准曲线以及载药量的测量
标准曲线的绘制:将1mg游离AQ4N溶于1mL去离子水中,完全溶解后,稀释配置为浓度为0.10、0.12、0.16、0.18、0.20mg/mL的溶液,用紫外分光光度计测量波长在618nm处吸光值,每个样本测量六次,作浓度(x)-吸光度(y)标准拟合曲线。巴诺蒽醌的标准曲线如图5所示。
紫外分光光度计检测中的AQ4N的载药率:取0.2mL AMMOFs(1mg/mL)加入含有0.2mL稀盐酸的反应瓶中,再添加0.8mL DMSO,充分使AMMOFs被还原释放药物,取7μL的溶液测量吸光值,测得的吸光度为0.253。根据标准曲线计算所测的AQ4N的含量,测得的药物含量按照以下公式计算载药率:
载药率(%)=W1/W2×100%
W1代表载AMMOFs内AQ4N的含量,W2代表AMMOFs的总量。
计算可知,实施例1制得的AMMOFs具有87.27%的高载药量。
上述实施例仅例示性说明本发明的原理及其功效,而非用于限制本发明。任何熟悉此技术的人士皆可在不违背本发明的精神及范畴下,对上述实施例进行修饰或改变。因此,举凡所属技术领域中具有通常知识者在未脱离本发明所揭示的精神与技术思想下所完成的一切等效修饰或改变,仍应由本发明的权利要求所涵盖。
Claims (10)
1.一种负载巴诺蒽醌的无载体纳米递送系统的制备方法,其特征在于,包括如下步骤:
(1)取巴诺蒽醌·二盐酸盐加入水中,搅拌溶解;
(2)向步骤(1)的溶液中逐滴滴加二氯化锰溶液,搅拌进行反应,再逐滴滴加三乙胺,搅拌反应,以去除HCl;
(3)将步骤(2)所得的反应液缓慢滴加至水中,搅拌进行反应,最后透析除去未配位的巴诺蒽醌,得到所述负载巴诺蒽醌的纳米递送系统。
2.根据权利要求1所述的制备方法,其特征在于:所述步骤(1)中,所述巴诺蒽醌·二盐酸盐的浓度为2mg/mL;
和/或,所述步骤(2)中,二氯化锰溶液的浓度为1mg/mL。
3.根据权利要求1所述的制备方法,其特征在于:所述步骤(1)中,搅拌时间为1-2h。
4.根据权利要求1所述的制备方法,其特征在于:所述步骤(2)中,巴诺蒽醌·二盐酸盐与三乙胺的用量比为1:(0.22-0.23)(mg/mL)。
5.根据权利要求1所述的制备方法,其特征在于:所述步骤(2)中,二氯化锰溶液加入后,继续搅拌反应3-5h;
和/或,所述步骤(2)中,三乙胺加入后,继续搅拌反应不低于8h。
6.根据权利要求1所述的制备方法,其特征在于:所述步骤(3)中,步骤(2)得到的反应液与水的体积用量比为1:4-6。
7.根据权利要求1所述的制备方法,其特征在于:所述步骤(3)中,搅拌反应时间为2-3h;
和/或,所述步骤(3)中,透析次数不低于3次,每次透析时间为不低于15min。
8.根据权利要求1所述的制备方法,其特征在于:所述步骤(1)、(2)、(3)中,反应全程需要在避光条件下进行;
和/或,所述步骤(1)、(2)、(3)都在室温下进行。
9.根据权利要求1所述的制备方法,其特征在于:所述纳米递送系统的粒径为118±22.86纳米。
10.一种根据权利要求1-9任一项所述的制备方法制得的负载巴诺蒽醌的无载体纳米递送系统。
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114344482A (zh) * | 2022-01-14 | 2022-04-15 | 重庆医科大学附属第二医院 | 一种基于金属有机骨架的多功能纳米粒及其制备方法与应用 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20130289520A1 (en) * | 2010-04-23 | 2013-10-31 | Children's Hospital Boston | Targeted and light-activated cytosolic drug delivery |
CN104837814A (zh) * | 2012-08-08 | 2015-08-12 | 拜欧斯戴特斯公司 | 新型化合物及其用途 |
CN106562930A (zh) * | 2016-10-24 | 2017-04-19 | 苏州大学 | 一种乏氧响应性脂质体制剂及其制备方法与应用 |
CN107007571A (zh) * | 2017-02-24 | 2017-08-04 | 福州市传染病医院 | 肿瘤微酸性敏感的铜‑药物共配位自组装纳米粒及应用 |
-
2021
- 2021-04-29 CN CN202110489072.7A patent/CN113368056A/zh active Pending
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20130289520A1 (en) * | 2010-04-23 | 2013-10-31 | Children's Hospital Boston | Targeted and light-activated cytosolic drug delivery |
CN104837814A (zh) * | 2012-08-08 | 2015-08-12 | 拜欧斯戴特斯公司 | 新型化合物及其用途 |
CN106562930A (zh) * | 2016-10-24 | 2017-04-19 | 苏州大学 | 一种乏氧响应性脂质体制剂及其制备方法与应用 |
CN107007571A (zh) * | 2017-02-24 | 2017-08-04 | 福州市传染病医院 | 肿瘤微酸性敏感的铜‑药物共配位自组装纳米粒及应用 |
Non-Patent Citations (5)
Title |
---|
DA ZHANG ET AL: "Chemotherapeutic Drug Based Metal–Organic Particles for Microvesicle-Mediated Deep Penetration and Programmable pH/NIR/Hypoxia Activated Cancer Photochemotherapy", 《ADVANCED SCIENCE》 * |
DA ZHANG ET AL: "pH/hypoxia programmable triggered cancer photo-chemotherapy based on a semiconducting polymer dot hybridized mesoporous silica framework", 《CHEMICAL SCIENCE》 * |
YU JI ET AL: "Tandem activated photodynamic and chemotherapy: Using pH-Sensitive nanosystems to realize different tumour distributions of photosensitizer/ prodrug for amplified combination therapy", 《BIOMATERIALS》 * |
张孝明: "Mn-DOX 金属—有机纳米颗粒的制备及其抗肿瘤诊疗一体的研究", 《中国优秀博硕士学位论文全文数据库(硕士)工程科技Ⅰ辑》 * |
梁金来等: "改善肿瘤乏氧环境及乏氧应激释药型抗肿瘤药纳米递送系统研究进展", 《中国药科大学学报》 * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114344482A (zh) * | 2022-01-14 | 2022-04-15 | 重庆医科大学附属第二医院 | 一种基于金属有机骨架的多功能纳米粒及其制备方法与应用 |
CN114344482B (zh) * | 2022-01-14 | 2023-05-12 | 重庆医科大学附属第二医院 | 一种基于金属有机骨架的多功能纳米粒及其制备方法与应用 |
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