CN107007569B - 一种载槲皮素和白藜芦醇的磁性脂质纳米粒及其制备方法 - Google Patents
一种载槲皮素和白藜芦醇的磁性脂质纳米粒及其制备方法 Download PDFInfo
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- CN107007569B CN107007569B CN201710178063.XA CN201710178063A CN107007569B CN 107007569 B CN107007569 B CN 107007569B CN 201710178063 A CN201710178063 A CN 201710178063A CN 107007569 B CN107007569 B CN 107007569B
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- quercetin
- resveratrol
- water
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- lipid
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Abstract
本发明涉及一种载槲皮素和白藜芦醇的磁性脂质纳米粒,其由以下重量份的组分组成:槲皮素1~4份、白藜芦醇1~4份、固体脂质20~100份、液态脂质40~100份、磷脂20~150份、水溶性乳化剂10~80份、磁性粒子4~15份、水2000~5000份。本发明还公开了该磁性脂质纳米粒的制备方法,可分别通过高温乳化‑低温固化、薄膜分散、乳化‑高压均质或微乳法制得。本发明的制备方法操作简便、清洁安全、无有毒有机溶剂残留,且可连续化生产,制得的磁性脂质纳米粒形态均一、粒径小、包封率高、稳定性好,实现了槲皮素和白藜芦醇的靶向载运与缓释,进一步增强了槲皮素和白藜芦醇两药合用的抗肿瘤作用。
Description
技术领域
本发明涉及一种载槲皮素和白藜芦醇的磁性脂质纳米粒及其制备方法,属于医药制剂技术领域。
背景技术
槲皮素(Quercetin,QT)属黄酮类化合物,具有舒张血管、降低血脂、抗氧化等多种生物活性及药理作用,近年来有关槲皮素抗肿瘤作用的研究亦日益引起重视。白藜芦醇(Resveratrol,Res)是广泛存在于多种食用和药用植物中的多酚类化合物。近年的研究表明,白藜芦醇抗肿瘤活性突出,可诱导肝癌、肺癌、乳腺癌、白血病等多种肿瘤细胞发生凋亡,被认为是最具希望的天然防癌剂之一。然而,槲皮素几乎不溶于水,在水中的溶解度仅有2.2μg/mL,白藜芦醇化学性质不稳定,见光遇热易分解,且水溶性差,在水中的溶解度仅为30μg/mL,因此槲皮素和白藜芦醇均存在口服吸收性差、生物利用度低等缺点,导致其临床应用受到一定程度的限制。
为提高槲皮素和白藜芦醇的生物利用度,人们已经研发出了一些药物载体结构,如专利CN 1850070A于2006年10月25日公开了一种槲皮素固体脂质纳米粒制剂,专利CN101982168A于2011年3月2日公开了一种槲皮素纳米胶束制剂,专利CN 104172184A于2014年12月3日公开了一种槲皮素纳米结构脂质载体,专利CN 105106117A于2015年12月2日公开了一种槲皮素的纳米粒,专利CN102614091A于2012年8月1日公开了一种白藜芦醇纳米结构脂质载体,专利CN103040754A于2013年4月17日公开了一种白藜芦醇纳米脂质体,专利CN104688715A于2015年6月10日公开了一种白藜芦醇固体脂质纳米粒,专利CN105534724A于2016年5月4日公开了一种包覆白藜芦醇的纳米固体脂质载体,等。以上制剂仅载有单一药物,虽能不同程度的提高槲皮素或白藜芦醇的生物利用度,但并不能同时发挥两药的药效,肿瘤靶向作用亦不明显。
发明内容
本发明的目的是提供一种载槲皮素和白藜芦醇的磁性脂质纳米粒,该磁性脂质纳米粒稳定性好、生物利用度高、肿瘤靶向性好,并且能够通过两药合用协同发挥抗肿瘤疗效。
本发明的另一目的是提供上述载槲皮素和白藜芦醇的磁性脂质纳米粒的制备方法。
本发明是通过以下技术方案实现的:一种载槲皮素和白藜芦醇的磁性脂质纳米粒,由以下重量份的组分组成:槲皮素1~4份、白藜芦醇1~4份、固体脂质20~100份、液态脂质40~100份、磷脂20~150份、水溶性乳化剂10~80份、磁性粒子4~15份、水2000~5000份;其中,
所述的固体脂质为单硬脂酸甘油酯、硬脂酸、软脂酸的一种或两种以上的混合物,
所述的液态脂质为辛酸癸酸甘油三酯、三油酸甘油酯、中链脂肪酸甘油三酯、肉豆蔻酸异丙酯、二乙二醇单乙基醚、油酸中的一种,
所述的磷脂为蛋黄卵磷脂或大豆卵磷脂,
所述的水溶性乳化剂为泊洛沙姆、胆酸钠中的一种或两种,
所述的磁性粒子为纳米四氧化三铁或油酸改性纳米四氧化三铁。
脂质纳米粒具有载药量理想、稳定性好、可大规模生产的特点,其脂质材料的组成是影响载药量和药物泄露的重要因素,本发明所述的固体脂质优选为单硬脂酸甘油酯和硬脂酸的混合物,所述单硬脂酸甘油酯和硬脂酸的质量比为5:1~9:1。
固体脂质会形成排列整齐的脂质晶格,限制纳米粒的载药能力,而液态脂质的引入能够增加载体的晶体混乱度,使其不易结晶,有利于提高载药量,降低药物在储存过程中的泄漏,本发明所述的液态脂质优选为辛酸癸酸甘油三酯。
乳化剂的种类可影响纳米粒的粒径、稳定性等,纳米载体包载药物和磁性材料得到磁靶向纳米给药系统,通过外磁场定位可选择性使药物定位于肿瘤组织。
本发明优选条件下提供的磁性脂质纳米粒特征为:电镜下呈类球形粒子,颗粒间分散、独立,平均粒径112nm,Zeta电位-27.78mV,白藜芦醇包封率为96.11%,槲皮素包封率为92.67%。
本发明的磁性脂质纳米粒可采用以下不同制备方法中的任意一种制得:
(1)高温乳化-低温固化法:按比例分别称取槲皮素、白藜芦醇、固体脂质、液态脂质和磷脂,溶解于有机溶剂中,构成油相;水溶性乳化剂和磁性粒子分散于水中,构成水相;在60~70℃条件下将油相缓慢滴加至水相,恒温持续搅拌至除尽有机溶剂,得初乳;将制得的初乳置冰水浴中搅拌,冷却后,超声探头乳化,即得载槲皮素和白藜芦醇的磁性脂质纳米粒;4℃保存或向制备的磁性脂质纳米粒中加入冻干保护剂甘露醇,冷冻干燥得固体制剂。
(2)高温乳化-低温固化法:按比例分别称取槲皮素、白藜芦醇、固体脂质、液态脂质和磷脂,溶解于有机溶剂中,构成油相;水溶性乳化剂分散于水中,构成水相;在60~70℃条件下将油相缓慢滴加至水相,恒温持续搅拌至除尽有机溶剂,得初乳;取水分散的磁性粒子,与初乳等体积混合后置冰水浴中搅拌,冷却后,超声探头乳化,即得载槲皮素和白藜芦醇的磁性脂质纳米粒;4℃保存或向制备的磁性脂质纳米粒中加入冻干保护剂甘露醇,冷冻干燥得固体制剂。
(3)薄膜分散法:按比例分别称取槲皮素、白藜芦醇、固体脂质、液态脂质和磷脂,溶解于有机溶剂中,构成油相,将油相转移至圆底烧瓶中,旋转蒸发器减压蒸发除去有机溶剂,使含药脂质在器壁形成薄膜,加入含水溶性乳化剂和磁性粒子的水相洗脱,将洗脱的混悬液进行超声探头乳化,即得载槲皮素和白藜芦醇的磁性脂质纳米粒;4℃保存或向制备的磁性脂质纳米粒中加入冻干保护剂甘露醇,冷冻干燥得固体制剂。
(4)乳化-高压均质法:按比例分别称取槲皮素、白藜芦醇、固体脂质、液态脂质和磷脂,溶解于有机溶剂中,构成油相;将水溶性乳化剂分散于水中,构成水相;在60~70℃条件下将水相一次性快速加入油相,高速搅拌形成初乳,迅速倒入高压均质机,1000bar压力下循环6次;冷却至室温,加入磁性粒子,搅拌,即得载槲皮素和白藜芦醇的磁性脂质纳米粒;4℃保存或向制备的磁性脂质纳米粒中加入冻干保护剂甘露醇,冷冻干燥得固体制剂。
(5)微乳法:按比例分别称取固体脂质、液态脂质和磷脂,热熔后构成油相,将槲皮素、白藜芦醇溶解于有机溶剂中,与油相混合均匀,搅拌至除去有机溶剂;称取水溶性乳化剂、磁性粒子分散于水中,构成水相;在60~70℃条件下将水相缓缓滴加到油相中,搅拌均匀,即得载槲皮素和白藜芦醇磁性脂质纳米粒;4℃保存或向制备的磁性脂质纳米粒中加入冻干保护剂甘露醇,冷冻干燥得固体制剂。
上述任一制备方法中,所述的有机溶剂为无水乙醇和丙酮的混合溶液,所述无水乙醇和丙酮的体积比为0.25:1~4:1。
本发明提供的磁性脂质纳米粒利用脂质纳米载体的疏水内核同时包载槲皮素和白藜芦醇,并将超顺磁纳米四氧化三铁载入含药脂质纳米粒,制成含药磁性脂质纳米粒,有效提高了槲皮素和白藜芦醇的溶解度和生物利用度,同时还可通过外磁场定位实现肿瘤靶向治疗,使药物能够定向浓集于靶区并释放,从而进一步增强了槲皮素和白藜芦醇两药合用的抗肿瘤效果,并降低药物在正常组织中的分布和聚集,减小其毒副作用。
本发明的制备方法多种多样、操作简便、清洁安全、无有毒有机溶剂残留,且可连续化生产,制得的磁性脂质纳米粒形态均一、粒径小、包封率高、稳定性好,实现了槲皮素和白藜芦醇的靶向载运与缓释。
附图说明
图1是超顺磁纳米四氧化三铁透射电镜图;
图2是油酸改性纳米四氧化三铁透射电镜图;
图3是载槲皮素和白藜芦醇的磁性脂质纳米粒透射电镜图;
图4是载槲皮素和白藜芦醇的磁性脂质纳米粒溶液高效液相色谱图;
图5是载槲皮素和白藜芦醇的磁性脂质纳米粒体外释放曲线图;
图6是载槲皮素和白藜芦醇的磁性脂质纳米粒磁滞回线图;
图7是槲皮素裸药、白藜芦醇裸药以及载槲皮素和白藜芦醇的磁性脂质纳米粒对HepG2肝癌细胞的抑制效果图。
具体实施方式
下面结合附图和具体实施例对本发明作进一步详细说明。
超顺磁纳米四氧化三铁的制备实验如下:
分取0.1mol/L的FeCl2·4H2O和FeCl3溶液各60ml,转移至250ml三颈烧瓶中。在体系温度50℃,1000rpm机械搅拌和氮气保护下,缓慢滴加25%的氨水溶液至pH值为12,搅拌反应30min后,冷却至室温,用水和无水乙醇反复洗涤生成物至近中性,磁吸后100℃真空干燥,即得超顺磁纳米四氧化三铁粒子。取适量四氧化三铁粒子的水分散液滴加到覆盖有碳膜的铜网上,自然晾干后,透射电镜下观察形态,透射电镜图如图1所示,粒子大小约5~10nm。
油酸改性纳米四氧化三铁的制备实验如下:
称取上述制备的超顺磁纳米四氧化三铁粒子0.5mg,均匀分散于水中,在体系温度60℃,1000rpm机械搅拌和氮气保护下,逐滴加入15ml油酸,搅拌反应30min,反应停止后,溶液冷却至室温,用无水乙醇洗涤油酸表面改性的Fe3O4至中性,磁吸分离后100℃真空干燥,即得油酸改性纳米四氧化三铁粒子。取适量油酸改性纳米四氧化三铁粒子的水分散液滴加到覆盖有碳膜的铜网上,自然晾干后,透射电镜下观察形态,透射电镜图如图2所示,粒子大小约10nm。
实施例1:精密称取槲皮素4.0mg、白藜芦醇4.0mg、单硬脂酸甘油酯50mg、硬脂酸10mg、辛酸癸酸甘油三酯40mg、蛋黄卵磷脂100mg,溶解于2.5ml无水乙醇和丙酮的混合溶剂(体积比1:1)中,70℃水浴下溶解作为油相;精密称取胆酸钠35mg、纳米Fe3O4 10mg,用5ml水分散作为水相;70℃、1000rpm机械搅拌下,将油相缓缓滴加到水相中,恒温持续搅拌2h除尽有机溶剂,得初乳;将制得的初乳置冰水浴中,800rpm搅拌30min,以300W的功率超声探头乳化6min,即得载槲皮素和白藜芦醇磁性脂质纳米粒。4℃保存或向制备的磁性脂质纳米粒中加入适量的甘露醇,冷冻干燥得固体制剂。
实施例2:
精密称取槲皮素2.5mg、白藜芦醇2.5mg、单硬脂酸甘油酯60mg、硬脂酸40mg、辛酸癸酸甘油三酯80mg、蛋黄卵磷脂100mg,溶解于2.5ml无水乙醇和丙酮的混合溶剂(体积比2:1)中,60℃水浴下溶解作为油相;精密称取泊洛沙姆50mg,用2.5ml水溶解作为水相;在60℃恒温条件及磁力搅拌下(1000rpm),将油相缓缓滴加到水相中,继续搅拌2h除尽有机溶剂,即得初乳。精密称取纳米Fe3O415mg,量取与初乳等量的水分散后,与初乳等体积混合,置冰浴下搅拌固化2h,以300W的功率超声探头乳化6min,即得载槲皮素和白藜芦醇磁性脂质纳米粒。
实施例3:
精密称取槲皮素2.5mg、白藜芦醇2.5mg、中链脂肪酸甘油酯100mg、硬脂酸50mg、单硬脂酸甘油酯50mg、大豆卵磷脂100mg,溶解于5ml无水乙醇和丙酮的混合溶剂(体积比0.5:1)中,70℃水浴下溶解作为油相;精密称取胆酸钠35mg,油酸改性纳米Fe3O4 15mg,用2.5ml水分散作为水相;在70℃恒温条件及机械搅拌下(1000rpm),将油相缓缓滴加到水相中,继续搅拌2h除尽有机溶剂,将其分散到适量的水中,使其总体积为5ml,冰水浴下搅拌30min,以300W的功率超声探头乳化6min,即得载槲皮素和白藜芦醇磁性脂质纳米粒。
实施例4:
精密称取槲皮素3.0mg、白藜芦醇3.0mg、单硬脂酸甘油酯90mg、硬脂酸10mg、肉豆蔻酸异丙酯40mg、大豆卵磷脂100mg,溶解于2ml无水乙醇和丙酮的混合溶剂(体积比4:1)中,70℃水浴下溶解作为油相;精密称取泊洛沙姆25mg、胆酸钠25mg,油酸改性纳米Fe3O410mg,用2.5ml水分散作为水相;在70℃恒温条件及机械搅拌下(1000rpm),将油相缓缓滴加到水相中,继续搅拌2h除尽有机溶剂,将其分散到适量的水中,使其总体积为5ml,冰水浴下搅拌30min,以300W的功率超声探头乳化6min,即得载槲皮素和白藜芦醇磁性脂质纳米粒。
实施例5:
精密称取槲皮素2.0mg、白藜芦醇2.0mg、三油酸甘油酯80mg、软脂酸20mg、大豆卵磷脂150mg,溶解于5ml无水乙醇和丙酮的混合溶剂(体积比0.25:1)中,60℃水浴下溶解作为油相;将油相转移至250ml圆底烧瓶中,旋转蒸发器减压蒸发除去有机溶剂,使含槲皮素和白藜芦醇的脂质在器壁形成一层薄膜,加入含胆酸钠35mg和纳米Fe3O410mg的5ml水相进行洗脱,洗脱的混悬液转移至西林瓶中,以300W的功率超声探头乳化6min,即得载槲皮素和白藜芦醇的磁性脂质纳米粒。
实施例6:
称取槲皮素50mg、白藜芦醇50mg、单硬脂酸甘油酯1g、辛酸癸酸甘油三酯2g、大豆卵磷脂1g,溶解于5ml无水乙醇和丙酮的混合溶剂(体积比1:1)中,构成油相;称取泊洛沙姆500mg,在70℃下用100ml水分散作为水相;将水相一次性快速加入到油相中,高速搅拌(6000rpm,3min)制成初乳,迅速倒入Panda Plus2000型高压均质机,1000bar压力下循环6次(每次10min),冷却至室温,加入纳米Fe3O4 200mg,60℃、1000rpm机械搅拌30min,即得载槲皮素和白藜芦醇的磁性脂质纳米粒。4℃保存或向制备的磁性脂质纳米粒中加入适量的甘露醇,冷冻干燥得固体制剂。
实施例7:
精密称取硬脂酸80mg、油酸60mg、大豆卵磷脂50mg热熔后作为油相,精密称取槲皮素2.0mg、白藜芦醇2.0mg,溶解于1ml无水乙醇和丙酮的混合溶剂(体积比1:1)后,与油相于60℃下混合均匀,1000rpm机械搅拌至除去有机溶剂;另精密称取泊洛沙姆80mg、纳米Fe3O48mg,用2.5ml水分散作为水相;在60℃恒温条件及机械搅拌下(1000rpm),将水相缓缓滴加到油相中,继续搅拌1h,即得载槲皮素和白藜芦醇的磁性脂质纳米粒。4℃保存或向制备的磁性脂质纳米粒中加入适量的甘露醇,冷冻干燥得固体制剂。
实施例8:
精密称取硬脂酸100mg、二乙二醇单乙基醚50mg、大豆卵磷脂50mg热熔后作为油相,精密称取槲皮素2.0mg、白藜芦醇2.0mg,溶解于1.5ml无水乙醇和丙酮的混合溶剂(体积比2:1)后,与油相于60℃下混合均匀,1000rpm机械搅拌至除去有机溶剂;另精密称取胆酸钠50mg、纳米Fe3O4 8mg,用2.5ml水分散作为水相;在60℃恒温条件及机械搅拌下(1000rpm),将水相缓缓滴加到油相中,继续搅拌1h,即得载槲皮素和白藜芦醇的磁性脂质纳米粒。4℃保存或向制备的磁性脂质纳米粒中加入适量的甘露醇,冷冻干燥得固体制剂。
通过以下试验(实施例1为例)对本发明的应用效果作进一步的说明:
1.形态观察
取载槲皮素和白藜芦醇的磁性脂质纳米粒水分散液少量滴于铜网上,自然干燥后,于Tecnai G2Spirit Twin型透射电镜下观察,为类球形粒子,粒径在20~180nm之间。结果见图3。
2.粒径与Zeta电位
取载槲皮素和白藜芦醇的磁性脂质纳米粒,用适量的水稀释,采用Nicomp380/ZLS型纳米粒度-Zeta电位测定仪测定其粒径及Zeta电位。Zeta电位值为-27.78mV,平均粒径为112nm,与透射电镜结果基本吻合。
3.包封率的测定
用超滤离心法-高效液相色谱法分离测定纳米粒中包封药物和游离药物,计算其包封率和载药量:取1.0ml纳米粒溶液置超滤离心管中,6500rpm离心20min,HPLC测定上清液中游离药物,计为W游;另取纳米粒溶液1ml,加甲醇适量,超声破乳,用甲醇定容至10ml,取适量分散液,离心,过滤,HPLC测定滤液中药物的含量,为W总。包封率(%)=(W总-W游)/W总×100。
液相色谱条件为:Shimadzu LC-20A高效液相色谱仪;色谱柱:Zorbax SB-C18色谱柱;流动相:甲醇-0.1%甲酸溶液;进样体积:20μl;柱温:30℃;流速:1ml/min;DAD检测器;检测波长:306nm(白藜芦醇)、360nm(槲皮素)。
HPLC图谱见图4。根据测定结果,计算得到白藜芦醇包封率为96.11%,槲皮素包封率为92.67%。
4.体外释放试验
精密量取纳米粒溶液装入透析袋(MWCO:3500),置于装有60mL 0.01mol/L磷酸盐缓冲液(PBS,pH7.4)溶液的烧杯中,保鲜膜封口。将装有透析袋的烧杯置于恒温气浴振荡器中(37℃,100rpm)考察其释放行为。分别于0.5、1、2、4、6、8、12、24、28h取1.5ml溶液,同时补充等量PBS;样品液用0.45μm微孔滤膜过滤,除去初滤液,取续滤液lml加入适量的流动相稀释,混匀,经高效液相色谱仪测定槲皮素和白藜芦醇含量,计算其累积释放百分率,绘制释放曲线。
载槲皮素和白藜芦醇的磁性脂质纳米粒体外释放曲线见图5,由图可知,12h内两种药物累积释放百分率达到80%以上,并具明显的缓释特征。
5.磁性特征
采用Lake Shore 7410型磁性超导量子磁强计测定载槲皮素和白藜芦醇的磁性脂质纳米粒的磁性特征,磁滞回线见图6。
6.肿瘤靶向性试验
荷H22肝癌细胞小鼠模型的建立:取昆明小鼠18只,体重15-20g,随即在每只小鼠右前侧腋下接种0.2ml的H22肝癌细胞悬液于皮下,接种后第3d,小鼠右侧腋下可触及移植局部有隆起的肿瘤硬块,接种第7d时肿瘤体积约100mm3,肿瘤形成。将成瘤小鼠随机分为3组,每组6只。给药方案分别为:生理盐水组、载槲皮素和白藜芦醇脂质纳米粒组(不含磁)、载槲皮素和白藜芦醇磁性脂质纳米粒组。给药方式为小鼠尾静脉注射,其中载槲皮素和白藜芦醇磁性脂质纳米粒组于小鼠肿瘤组织部位施加外磁场。小鼠尾静脉注射后,分别于0.5、4、12、24h取出动物,颈椎脱臼法处死小鼠,立刻进行解剖,取出心、肝、脾、肺、肾、肿瘤,在生理盐水中洗涮,滤纸吸干表面水分,称量记录组织湿重,取部分组织称重后加入等量蒸馏水进行组织匀浆,取组织匀浆液20μl,涡旋混合均匀,加入蛋白沉淀溶剂乙腈60μl,涡旋混合3min,然后再补充加入萃取剂,10000rpm离心10min,沉淀蛋白及脂质,HPLC测定上清液中药物的含量。
结果显示载槲皮素和白藜芦醇磁性脂质纳米粒组的荷瘤小鼠除肿瘤组织外其余组织中的分布量均较少;而载槲皮素和白藜芦醇脂质纳米粒(不含磁)组的荷瘤小鼠肿瘤组织和肝脏分布最多,再次是心、肾、脾、肺,充分证明磁性脂质纳米粒较不含磁的脂质纳米粒相比,具有更明显的肿瘤靶向性。
7.肿瘤细胞杀伤试验
HepG2肝癌细胞株在含10%胎牛血清、100U/ml青霉素、100μg/ml链霉素的DMEM培养液中,置于37℃,5%CO2培养箱中培养。采用CCK-8法测定肿瘤细胞活力。具体操作如下:将HepG2细胞悬液以1×105/ml的密度接种于96孔板,设对照组和实验组,每组3个复孔,以未处理的细胞作为对照组,实验组分设槲皮素、白藜芦醇、槲皮素+白藜芦醇、槲皮素磁性脂质纳米粒、白藜芦醇磁性脂质纳米粒、载槲皮素和白藜芦醇磁性脂质纳米粒,培养24h后,每孔加入20μl CCK-8溶液,继续培养2h,随后用BioTek ELx808型多功能酶标仪测定细胞在450nm处的吸光度(OD值),并按以下公式计算肿瘤细胞活力(%),各实验组对HepG2细胞的抑制效果见图7。
肿瘤细胞活力(%)=OD实验/OD对照×100%。
结果表明,载槲皮素和白藜芦醇的磁性脂质纳米粒对HepG2细胞具有明显的抑制作用,且载槲皮素和白藜芦醇的磁性脂质纳米粒对于肿瘤细胞的杀伤作用明显优于载槲皮素或载白藜芦醇单药的脂质纳米粒(P<0.05)。
Claims (8)
1.一种载槲皮素和白藜芦醇的磁性脂质纳米粒,其特征在于:由以下重量份的组分组成:槲皮素1~4份、白藜芦醇1~4份、固体脂质20~100份、液态脂质40~100份、磷脂20~150份、水溶性乳化剂10~80份、磁性粒子4~15份、水2000~5000份;其中,
所述的固体脂质为单硬脂酸甘油酯和硬脂酸的混合物,所述单硬脂酸甘油酯和硬脂酸的质量比为5:1~9:1,
所述的液态脂质为辛酸癸酸甘油三酯,
所述的磷脂为蛋黄卵磷脂或大豆卵磷脂,
所述的水溶性乳化剂为泊洛沙姆、胆酸钠中的一种或两种,
所述的磁性粒子为纳米四氧化三铁或油酸改性纳米四氧化三铁。
2.根据权利要求1所述的载槲皮素和白藜芦醇的磁性脂质纳米粒,其特征在于:所述磁性脂质纳米粒的粒径不大于200nm。
3.根据权利要求1所述的载槲皮素和白藜芦醇的磁性脂质纳米粒的制备方法,其特征在于:包括以下步骤:按比例分别称取槲皮素、白藜芦醇、固体脂质、液态脂质和磷脂,溶解于有机溶剂中,构成油相;水溶性乳化剂和磁性粒子分散于水中,构成水相;在60~70℃条件下将油相缓慢滴加至水相,恒温持续搅拌至除尽有机溶剂,得初乳;将制得的初乳置冰水浴中搅拌,冷却后,超声探头乳化,即得载槲皮素和白藜芦醇的磁性脂质纳米粒;4℃保存或向制备的磁性脂质纳米粒中加入冻干保护剂甘露醇,冷冻干燥得固体制剂。
4.根据权利要求1所述的载槲皮素和白藜芦醇的磁性脂质纳米粒的制备方法,其特征在于:包括以下步骤:按比例分别称取槲皮素、白藜芦醇、固体脂质、液态脂质和磷脂,溶解于有机溶剂中,构成油相;水溶性乳化剂分散于水中,构成水相;在60~70℃条件下将油相缓慢滴加至水相,恒温持续搅拌至除尽有机溶剂,得初乳;取水分散的磁性粒子,与初乳等体积混合后置冰水浴中搅拌,冷却后,超声探头乳化,即得载槲皮素和白藜芦醇的磁性脂质纳米粒;4℃保存或向制备的磁性脂质纳米粒中加入冻干保护剂甘露醇,冷冻干燥得固体制剂。
5.根据权利要求1所述的载槲皮素和白藜芦醇的磁性脂质纳米粒的制备方法,其特征在于:包括以下步骤:按比例分别称取槲皮素、白藜芦醇、固体脂质、液态脂质和磷脂,溶解于有机溶剂中,构成油相,将油相转移至圆底烧瓶中,旋转蒸发器减压蒸发除去有机溶剂,使含药脂质在器壁形成薄膜,加入含水溶性乳化剂和磁性粒子的水相洗脱,将洗脱的混悬液进行超声探头乳化,即得载槲皮素和白藜芦醇的磁性脂质纳米粒;4℃保存或向制备的磁性脂质纳米粒中加入冻干保护剂甘露醇,冷冻干燥得固体制剂。
6.根据权利要求1所述的载槲皮素和白藜芦醇的磁性脂质纳米粒的制备方法,其特征在于:包括以下步骤:按比例分别称取槲皮素、白藜芦醇、固体脂质、液态脂质和磷脂,溶解于有机溶剂中,构成油相;将水溶性乳化剂分散于水中,构成水相;在60~70℃条件下将水相一次性快速加入油相,高速搅拌形成初乳,迅速倒入高压均质机,1000bar压力下循环6次;冷却至室温,加入磁性粒子,搅拌,即得载槲皮素和白藜芦醇的磁性脂质纳米粒;4℃保存或向制备的磁性脂质纳米粒中加入冻干保护剂甘露醇,冷冻干燥得固体制剂。
7.根据权利要求1所述的载槲皮素和白藜芦醇的磁性脂质纳米粒的制备方法,其特征在于:包括以下步骤:按比例分别称取固体脂质、液态脂质和磷脂,热熔后构成油相,将槲皮素、白藜芦醇溶解于有机溶剂中,与油相混合均匀,搅拌至除去有机溶剂;称取水溶性乳化剂、磁性粒子分散于水中,构成水相;在60~70℃条件下将水相缓缓滴加到油相中,搅拌均匀,即得载槲皮素和白藜芦醇磁性脂质纳米粒;4℃保存或向制备的磁性脂质纳米粒中加入冻干保护剂甘露醇,冷冻干燥得固体制剂。
8.根据权利要求3至7任一项所述的载槲皮素和白藜芦醇的磁性脂质纳米粒的制备方法,其特征在于:所述的有机溶剂为无水乙醇和丙酮的混合溶液,所述无水乙醇和丙酮的体积比为0.25:1~4:1。
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Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1932517A2 (en) * | 2006-12-11 | 2008-06-18 | Universiteit Utrecht Holding B.V. | Liposomes containing a polyphenol derivative such as caffeic acid and a method of post-loading thereof |
CN105457038A (zh) * | 2015-11-09 | 2016-04-06 | 东南大学 | 一种速释型药物磷脂化合物及其药物组合物 |
WO2016053809A1 (en) * | 2014-09-29 | 2016-04-07 | Barrie Tan | Non-synthetic emulsion-based lipid formulations and methods of use |
-
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Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1932517A2 (en) * | 2006-12-11 | 2008-06-18 | Universiteit Utrecht Holding B.V. | Liposomes containing a polyphenol derivative such as caffeic acid and a method of post-loading thereof |
WO2016053809A1 (en) * | 2014-09-29 | 2016-04-07 | Barrie Tan | Non-synthetic emulsion-based lipid formulations and methods of use |
CN105457038A (zh) * | 2015-11-09 | 2016-04-06 | 东南大学 | 一种速释型药物磷脂化合物及其药物组合物 |
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