CN106267248A - 一种载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡及其制备方法 - Google Patents
一种载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡及其制备方法 Download PDFInfo
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Abstract
本发明公开一种载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡及其制备方法。所述脂质超声微泡包括磷脂膜,气体和载药的叶酸修饰介孔二氧化硅纳米粒。该新型微泡载药量高,制备工艺简单,生产成本低,易于工业化生产,不但可以作为超声显影剂用于疾病诊断,还可以作为载药制剂用于疾病治疗。
Description
技术领域
本文具体涉及一种载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡及其制备方法,属于药物领域。
背景技术
载气脂质超声微泡前后经历几代的发展,已有产品上市。近年的研究发现,微泡可在一定强度的超声辐照下破裂,产生一定的生物作用,这种现象和所产生的生物作用被应用于靶向给药系统,实现特定部位给药。通过不同的制剂技术,微泡可以携带不同的药物。微泡注射进入体内后,在外加的超声辐照下,可以在特定的部位实现超声诱导击破,释放其所携带的药物,从而实现靶向给药。但是微泡存在着载药量低和包封率低的问题,大大限制了其在临床上的应用。
纳米给药系统一直是靶向给药系统研究的热点。介孔二氧化硅纳米粒具有稳定性好,载药量高,生物相容性好和表面可修饰性等诸多优势,近年来受到了广泛的关注,成为了新的研究热点。部分肿瘤细胞表面高表达叶酸受体,而普通正常组织细胞表面叶酸受体表达则相对较低。在本发明中,通过本发明所阐述的方法,在介孔二氧化硅纳米粒的表面共价修饰叶酸分子,可以实现主动靶向肿瘤细胞的目的。本发明所制备的载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡注射进入体内后,可通过体液循环到达肿瘤部位。在超声成像引导下,通过超声辐照实现载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡在肿瘤目标部位的定向击破,随即释放其所携带的载药叶酸修饰介孔二氧化硅纳米粒。释放的载药叶酸修饰介孔二氧化硅纳米粒通过肿瘤部位的EPR效应进入到肿瘤间质后,载药叶酸修饰介孔二氧化硅纳米粒可通过其表面修饰的叶酸配体与肿瘤细胞表面的叶酸受体特异性结合,在叶酸受体介导的胞吞作用下,进入到肿瘤细胞内部,实现肿瘤细胞内部给药。
本发明所制备的载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡可以实现多重靶向作用,降低该药物毒副作用的同时,大大增强药物的治疗作用。
发明内容
本发明的目的在于针对微泡的低包封率和低载药量以及所载药物的毒副作用等问题,提供一种新的中间同时包载气体和载药叶酸修饰介孔二氧化硅纳米粒的脂质微泡及其制备方法。该新型载叶酸修饰介孔二氧化硅纳米粒的脂质微泡在提高微泡包封率和载药量的同时,还可在外加超声辐照下,击破微泡并释放其携带的载药叶酸修饰介孔二氧化硅纳米粒,从而实现多重靶向作用,降低该制剂毒性的同时,大大增强药物的治疗作用。
本发明的目的可以通过以下技术解决方案实现,但本发明并不仅限于下述技术:
步骤1:称取十六烷基三甲基溴化铵(CTAB)和氢氧化钠(NaOH)溶于80℃的去离子水中。吸取正硅酸乙酯(TEOS)滴加到上述溶液中,恒温搅拌反应3h。6%酸醇溶液(v/v)回流洗去模板剂CTAB,离心收集,得到介孔二氧化硅纳米粒。
步骤2:使用3-氨丙基三乙氧基硅烷(APTES)修饰步骤1制得的介孔二氧化硅纳米粒,得到氨基修饰介孔二氧化硅纳米粒。
步骤3:称取适量的叶酸(FA),N-N-羟基琥珀酰亚胺(NHS),二环己基碳二亚胺(DCC)溶于一定量的无水二甲基亚砜(DMSO),25℃避光搅拌反应24h,得到活化的叶酸活性脂。
步骤4:称取步骤2得到的氨基修饰介孔二氧化硅纳米粒和步骤3得到叶酸活性脂,分散于无水二甲基亚砜(DMSO),搅拌反应24h,得到叶酸修饰介孔二氧化硅纳米粒。
步骤5:制备药物乙醇饱和溶液,称取步骤4制得的叶酸修饰介孔二氧化硅纳米粒加入到上述药物乙醇溶液中,旋转蒸发除去乙醇,得到载药叶酸修饰介孔二氧化硅纳米粒。
步骤6:将二棕榈酰磷脂酰胆碱(DPPC),二棕榈酰磷酯酰乙醇胺(DPPE),甘油和磷酸盐缓冲液(PBS)按比例混合,称取步骤5得到的载药叶酸修饰介孔二氧化硅纳米粒,70℃孵化30min,冲入氟碳气剪切2min,得到载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡。
步骤7:香豆素6标记叶酸修饰介孔二氧化硅纳米粒,制备载香豆素6标记叶酸修饰介孔二氧化硅纳米粒脂质超声微泡,采用激光共聚焦显微镜观察其外观形貌。
附图说明
图1为本发明实例1所制备的介孔二氧化硅纳米粒(MSN),氨基修饰介孔二氧化硅纳米粒(MSN-NH2)和叶酸修饰介孔二氧化硅纳米粒(MSN-FA)的红外光谱图
图2为本发明实施例1所制备介孔二氧化硅纳米粒(A)和叶酸修饰介孔二氧化硅纳米粒(B)的透射图
图3为本发明实例1所制备的空白脂质超声微泡粒径分布图
图4为本发明实例1所制备的载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡的粒径分布图
图5为本发明实例1所制备的空白脂质超声微泡(MB)和载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡(MSN-FA-TAN-MB)的体外显影图
图6为本发明实例1所制备的空白脂质超声微泡(MB)和载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡(MSN-FA-TAN-MB)注射前(pre)后(post)的体内显影对比图
图7为本发明实例1所制备的载香豆素6标记叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡的激光共聚焦观察图
具体实施方式
以下结合附图实施例对本发明进行详细描述,但本发明并不仅限于下述实施例。
实施例1
1.介孔二氧化硅纳米粒的制备
分别称取1.0g CTAB和0.28g NaOH溶于80℃水中,吸取5mL TEOS滴加到上述溶液中,恒温机械搅拌,离心收集产物。酸醇溶液回流,离心洗涤收集产物,干燥,制得空白介孔二氧化硅纳米粒(MSN)。
2.氨基修饰介孔二氧化硅纳米粒的制备
称取0.4g空白介孔二氧化硅纳米粒加入到25mL无水乙醇中,超声分散,向上述溶液中滴加1.5mL的APTES,室温下搅拌反应24h后,无水乙醇洗涤数次,离心收集产品,干燥,得到氨基修饰介孔二氧化硅纳米粒(MSN-NH2)。
3.叶酸修饰介孔二氧化硅纳米粒的制备
分别取1.0g叶酸,0.52g NHS,0.94g DCC和0.5mL三乙胺溶于20mL无水DMSO中,25℃避光搅拌反应24h。反应结束后过滤,乙醚洗涤,干燥得到叶酸活性脂。
称取200mg氨基修饰介孔二氧化硅纳米粒和20mg叶酸活性脂,加入到20mL无水DMSO中,25℃避光搅拌反应24h。离心收集,洗涤,得到叶酸修饰介孔二氧化硅纳米粒。
将制备成的叶酸修饰介孔二氧化硅纳米粒以及前面制备的介孔二氧化硅纳米粒和氨基修饰介孔二氧化硅纳米粒真空干燥,红外检测,得到的红外光谱图见图1。三个样品均出现了介孔二氧化硅纳米粒的特征吸收峰,具有代表性的几个特征吸收峰如下:1076cm-1为Si-O-Si的伸缩振动峰,470cm-1为Si-O-Si的弯曲震动峰,960cm-1为Si-OH的伸缩振动峰。相比空白介孔二氧化硅纳米粒(MSN)的红外光谱图,氨基修饰介孔二氧化硅纳米粒(MSN-NH2)光谱图在2922cm-1和2855cm-1处出现了两个明显的新吸收峰,这两个吸收峰为氨基修饰介孔二氧化硅纳米粒丙基中的C-H伸缩振动峰,这两个新吸收峰的出现证明了氨基修饰介孔二氧化硅纳米粒的成功制备。另外,相比空白介孔二氧化硅纳米粒和氨基修饰介孔二氧化硅纳米粒的红外图谱,叶酸修饰介孔二氧化硅纳米粒(MSN-FA)在1650cm-1和1550cm-1处出现了新的吸收峰,1650cm-1和1550cm-1是酰胺键的特征吸收峰,它们的出现成功证明了氨基修饰介孔二氧化硅纳米粒的成功制备。
将制备成的叶酸修饰介孔二氧化硅纳米粒和空白介孔二氧化硅纳米粒制成合适浓度的水分散液,送检作透射显微镜观察,结果如图2。从图中可以看出,叶酸修饰介孔二氧化硅纳米粒和空白介孔二氧化硅纳米粒大小均一,形状呈类圆形,粒径分布在110nm左右。相比空白介孔二氧化硅纳米粒,叶酸修饰介孔二氧化硅纳米粒在形态和粒径等方面没有明显改变,说明表面叶酸修饰对介孔二氧化硅纳米粒形态和粒径影响较小。
4.载药叶酸修饰介孔二氧化硅纳米粒的制备
称取160mg叶酸修饰介孔二氧化硅纳米粒和40mg丹参酮II A分散于40mL的无水乙醇中,超声分散,旋转蒸发除去乙醇,洗去多余的药物,得到载药叶酸修饰介孔二氧化硅纳米粒。
5.载叶酸修饰介孔二氧化硅纳米粒脂质超声微泡的制备
称取5mg DPPC,2mg DPPE和一定量的载药叶酸修饰介孔二氧化硅纳米粒分散在含有0.4mL甘油的4mL PBS中,70℃孵化30min。然后剪切2min,剪切的过程中通入5mL氟碳气,30s内通完,PBS离心洗涤,得到载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡。
将制备成的载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡混悬液和空白脂质超声稀释10倍,采用马尔文激光粒度分析仪测量其粒径。粒径分布图见图3和4,相比空白脂质超声微泡粒径(2061±442.6nm),载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡粒径(2608±605.6nm)有所增大,推测为由于载药叶酸修饰介孔二氧化硅纳米粒的包载,一定程度上增大了载叶酸修饰介孔二氧化硅纳米粒脂质超声微泡的粒径。
6.体外显影
将制备的载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡注入到自制的凝胶模型中,测定其显影效果,结果见图5。结果表明本发明制备的载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡具有良好的体外增强显影效果。
7.体内显影
体外培养H22细胞(由重庆市生物化学与分子药理重点实验室提供),培养液稀释注入昆明鼠(由重庆医科大学动物中心提供)的腹部,待其长成腹水,传至5代。吸取腹水,生理盐水稀释,皮下注射到新领昆明鼠的腋下,建立H22荷瘤小鼠模型。
以建成的H22荷瘤小鼠模型为实验对象,采用自身前后对照法,采集H22荷瘤小鼠肿瘤造影前后的超声响应图像,对比评价本发明制备的载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡的显影效果。
具体操作如下:将0.2mL本发明制备的载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡经尾静脉注入H22荷瘤小鼠的体内,观察本发明制备的载叶酸修饰介孔二氧化硅纳米粒脂质超声微泡体内显影效果。结果(见图6)表明本发明制备的制备的载叶酸修饰介孔二氧化硅纳米粒脂质超声微泡具有良好的体内增强显影效果。
8.激光共聚焦显微镜观察
采用香豆素6标记叶酸修饰介孔二氧化硅纳米粒,将制备好的载香豆素6标记叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡置于激光共聚焦显微镜下观察,结果见图7。由图可见,叶酸修饰介孔二氧化硅纳米粒被成功的包载进脂质超声微泡。
Claims (6)
1.一种载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡,其特征在于:所述脂质超声微泡包括磷脂膜,气体和载药的叶酸修饰介孔二氧化硅纳米粒。
2.如权利要求1所述的载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡,其特征在于所述制剂中各组分重量百分比为:
溶剂为甘油,水,磷酸盐缓冲液或有机溶剂;
所述的有机溶剂为乙醇,二甲亚砜或三氯甲烷;
所述的载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡的步骤如下:(a)称取处方量的叶酸修饰介孔二氧化硅纳米粒和处方量的药物,采用溶剂挥发法或吸附法方法实现叶酸修饰介孔二氧化硅纳米粒的载药,得到载药叶酸修饰介孔二氧化硅纳米粒;(b)将磷脂成分加入热的甘油和水或磷酸盐缓冲液的混合溶液中孵化;(c)将步骤(a)中制得的载药叶酸修饰介孔二氧化硅纳米粒加入步骤(b)中制得的磷脂混合溶液中孵化;(d)孵化后,将所载的气体充入载药容器中,利用超声、剪切、震荡或冻干方式促使溶液形成粒径均匀且物理性质稳定的乳状液。即得到载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡。
3.如权利要求1所述的载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡,其特征在于:所述磷脂为二棕榈酰磷脂酰胆碱,二棕榈酰磷酯酰乙醇胺,二硬脂酰磷脂酰乙醇胺,二硬脂酰磷脂酰胆碱,二月桂酰磷脂酰胆碱,二肉豆蔻酰磷脂酰胆碱,二花生酰基磷脂酰胆碱,二油酰磷脂酰胆碱,双十五酰基-磷脂酰胆碱,1-肉豆蔻酰-2-棕榈酰-磷脂酰胆碱,1-棕榈酰-2-肉豆蔻酰-磷脂酰胆碱,1-棕榈酰-2-硬脂酰-磷脂酰胆碱,1-硬脂酰-2-棕榈酰-磷脂酰胆碱,1-棕榈酰-2-油基-磷脂酰胆碱,1-油酰-2-棕榈酰-磷脂酰胆碱及药学上可接受的天然磷脂中的一种或几种混合。
4.如权利要求1所述载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡,其特征在于:所述的气体选自氟碳气体、空气、氧气、氮气和二氧化碳其中的一种或几种混合的气体。
5.如权利要求1所述的载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡,其特征在于:包载于叶酸修饰介孔二氧化硅纳米粒内部的药物可以是:药物;生物活性剂;细胞毒性剂;化疗剂;放射治疗剂;蛋白质;天然或合成的肽,包括寡肽和多肽;维生素;类固醇;遗传物质,包括核苷,核苷酸,寡核苷酸,多核苷酸和质粒。
6.如权利要求2所述的载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡,其特征在于:脂质超声微泡的制备方法可以为超声法,剪切法,震荡法或冻干法。
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Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107213476A (zh) * | 2017-07-12 | 2017-09-29 | 重庆医科大学 | 一种透明质酸修饰的硅包裹载药磷脂液态氟碳纳米球超声造影剂及其制备方法 |
CN109675034A (zh) * | 2019-02-22 | 2019-04-26 | 重庆医科大学 | 一种多模态造影剂及其用途 |
CN113384530A (zh) * | 2020-03-13 | 2021-09-14 | 暨南大学 | 一种多糖核心Nanocells及其制备方法与应用 |
CN114146188A (zh) * | 2021-12-14 | 2022-03-08 | 河南工业大学 | 一种修饰型LMSNs纳米药物载体的制备方法 |
CN115227833A (zh) * | 2022-07-29 | 2022-10-25 | 西安交通大学医学院第一附属医院 | 一种氟化二氧化硅载药纳米粒及其制备方法、用途 |
CN116327985A (zh) * | 2023-02-21 | 2023-06-27 | 武汉工程大学 | 一种具有靶向超声造影成像功能的空心囊泡载药递送系统及其制备方法与应用 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103990130A (zh) * | 2013-06-24 | 2014-08-20 | 中国人民解放军第二军医大学 | 介孔二氧化硅纳米制剂及其制备方法和应用 |
CN104096245A (zh) * | 2014-07-18 | 2014-10-15 | 重庆医科大学 | 包裹载药白蛋白纳米粒的脂质超声微泡及其制备方法 |
-
2016
- 2016-09-08 CN CN201610815688.8A patent/CN106267248B/zh not_active Expired - Fee Related
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103990130A (zh) * | 2013-06-24 | 2014-08-20 | 中国人民解放军第二军医大学 | 介孔二氧化硅纳米制剂及其制备方法和应用 |
CN104096245A (zh) * | 2014-07-18 | 2014-10-15 | 重庆医科大学 | 包裹载药白蛋白纳米粒的脂质超声微泡及其制备方法 |
Non-Patent Citations (1)
Title |
---|
HENGLI YANG ET AL.: "NanobubbleeAffibody: Novel ultrasound contrast agents for targeted molecular ultrasound imaging of tumor", 《BIOMATERIALS》 * |
Cited By (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107213476A (zh) * | 2017-07-12 | 2017-09-29 | 重庆医科大学 | 一种透明质酸修饰的硅包裹载药磷脂液态氟碳纳米球超声造影剂及其制备方法 |
CN107213476B (zh) * | 2017-07-12 | 2021-01-19 | 重庆医科大学 | 一种透明质酸修饰的硅包裹载药磷脂液态氟碳纳米球超声造影剂及其制备方法 |
CN109675034A (zh) * | 2019-02-22 | 2019-04-26 | 重庆医科大学 | 一种多模态造影剂及其用途 |
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