CN113368054A - 一种多聚脱氧核糖核苷酸柔性纳米脂质体的制备方法 - Google Patents
一种多聚脱氧核糖核苷酸柔性纳米脂质体的制备方法 Download PDFInfo
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Abstract
一种多聚脱氧核糖核苷酸柔性纳米脂质体的制备方法,本发明涉及脂质体制备技术领域,取大豆卵磷脂、胆固醇和聚氧乙烯脱水山梨醇单油酸酯溶解于乙醚溶剂中,恒温30‑40℃减压旋转蒸发掉有机溶剂,使大豆卵磷脂在蒸发溶器底形成一层薄膜后,加入乙醚溶剂与磷酸盐缓冲液调节pH5‑8,继续旋转蒸发水合20‑40分钟后,用0.45微米滤膜过滤后得到空白脂质体溶液;向空白脂质体溶液中加入PDRN、胆酸钠,混合均匀后,进行超声3‑5分钟,加入适量的磷酸盐缓冲液,继续旋转蒸发水合20‑40分钟后,过滤后得到PDRN柔性纳米脂质体。可以显著促进PDRN的透皮吸收并提高其作用效率,还能提高PDRN在皮肤表面和皮肤深层的滞留和作用时间;粉末状态的PDRN柔性纳米脂质体还利于使用、运输和贮存。
Description
技术领域
本发明涉及脂质体制备技术领域,具体涉及一种多聚脱氧核糖核苷酸柔性纳米脂质体的制备方法。
背景技术
皮肤是人体的第一道对外的保护层,补水是护肤中的首要任务,因为肌肤的水分非常容易流失,而肌肤缺乏足够的含水量将直接导致肌肤代谢不正常而衰老、出现皱纹和色斑等等一系列肌肤问题。然而,空调、环境污染、季节转换带来的温度变化与压力,以及皮肤衰老、代谢减缓等因素都会造成肌肤水分流失。一般来说皮肤的保湿就是皮肤的持水能力,能够减缓皮肤内水分蒸发流失的速度。
多聚脱氧核糖核苷酸(简称:PDRN)作为生物体的重要遗传物质,一方面其在调控基因、蛋白表达,改善细胞状态,维持机体的正常生理功能方面发挥着重要的作用;另一方面它作为原料库,为机体生长、发育、修复提供了所需要的脱氧核糖核苷酸。但由于PDRN链比较长,分子量大且带有电荷,导致其不宜通过细胞膜被机体吸收。制药、医美、保健品行业的科研工作者长期致力于寻找碱基组成与人体相似性高的DNA生物材料。经过研究者不懈的努力,证实三文鱼DNA的碱基组成与人体DNA的相似性达98%。
因此,PDRN直接作用于炎症愈合、损伤修复,由内而外加筑肌肤屏障,复原细胞活性,加速细胞新陈代谢,提高皮肤修复和再生能力,重建健康肌底,但PDRN的脂溶性不高,生物利用度低,所以需要改进。
发明内容
本发明的目的在于针对现有技术的缺陷和不足,提供了一种多聚脱氧核糖核苷酸柔性纳米脂质体的制备方法,可以显著促进PDRN的透皮吸收并提高其作用效率,还能提高PDRN在皮肤表面和皮肤深层的滞留和作用时间;粉末状态的PDRN柔性纳米脂质体还利于使用、运输和贮存;可采用机械化方法制备,产品工艺和质量的稳定性好、重现性高,广泛用于制药、医美、保健品以及食品行业中。
为达到上述目的,本发明采用了下列技术方案:它的操作步骤如下:
步骤一、空白脂质体的制备:
取大豆卵磷脂、胆固醇和聚氧乙烯脱水山梨醇单油酸酯溶解于乙醚溶剂中,其中,大豆卵磷脂重量配比为150-250份,胆固醇重量配比为30-90份,聚氧乙烯脱水山梨醇单油酸酯重量配比为20-100份,恒温30-40℃减压旋转蒸发掉有机溶剂,使大豆卵磷脂在蒸发溶器底形成一层薄膜后,加入乙醚溶剂与磷酸盐缓冲液体积比从1∶1到1∶5,调节pH5-8,继续旋转蒸发水合20-40分钟后,用0.45微米滤膜过滤后得到空白脂质体溶液;
步骤二、PDRN柔性纳米脂质体的制备:
向空白脂质体溶液中加入PDRN重量配比为1-40份、胆酸钠20-40份,混合均匀后,进行超声3-5分钟,加入适量的磷酸盐缓冲液,调节pH5-8,继续旋转蒸发水合20-40分钟后,用0.45微米滤膜过滤后得到PDRN柔性纳米脂质体,最后冷处放置保存。
优选地,所述PDRN柔性纳米脂质体为球形单室脂质体,形态均一,包封率高达95%以上,粒子直径平均分布在150nm以下。
优选地,所述PDRN柔性纳米脂质体中含有PDRN重量配比为5-40份。
优选地,所述磷酸盐缓冲液调节pH值为7.4。
优选地,所述PDRN柔性纳米脂质体应用于制药、医美、保健品以及食品行业中。
与现有技术相比,本发明的有益效果是:本发明提供了一种多聚脱氧核糖核苷酸柔性纳米脂质体的制备方法,可以显著促进PDRN的透皮吸收并提高其作用效率,还能提高PDRN在皮肤表面和皮肤深层的滞留和作用时间;粉末状态的PDRN柔性纳米脂质体还利于使用、运输和贮存;可采用机械化方法制备,产品工艺和质量的稳定性好、重现性高,广泛用于制药、医美、保健品以及食品行业中。
具体实施方式:
本具体实施方式(实施例一)采用如下技术方案:本实施例的操作步骤如下:
步骤一、空白脂质体的制备:
取大豆卵磷脂150g、胆固醇30g和聚氧乙烯脱水山梨醇单油酸酯20g溶解于适量乙醚溶剂中,恒温30-40℃减压旋转蒸发掉有机溶剂,使大豆卵磷脂在蒸发溶器底形成一层薄膜后,加入剩余乙醚溶剂与160ml的磷酸盐缓冲液调节pH7.4,继续旋转蒸发水合20-40分钟后,用0.45微米滤膜过滤后得到空白脂质体溶液,上述乙醚溶剂工40ml;
步骤二、PDRN柔性纳米脂质体的制备:
向空白脂质体溶液中加入PDRN1g、胆酸钠20g,混合均匀后,进行超声3-5分钟,加入100ml的磷酸盐缓冲液,调节pH7.4,继续旋转蒸发水合20-40分钟后,用0.45微米滤膜过滤后得到PDRN柔性纳米脂质体,最后冷处放置保存;
步骤三、采用超速离心法(转速:15000r/min,),离心20min,测定PDRN柔性纳米脂质体的包封率高达95.8%。
本具体实施方式的有益效果是:本具体实施方式提供了一种多聚脱氧核糖核苷酸柔性纳米脂质体的制备方法,可以显著促进PDRN的透皮吸收并提高其作用效率,还能提高PDRN在皮肤表面和皮肤深层的滞留和作用时间;粉末状态的PDRN柔性纳米脂质体还利于使用、运输和贮存;可采用机械化方法制备,产品工艺和质量的稳定性好、重现性高,广泛用于制药、医美、保健品以及食品行业中。
实施例二:
本实施例的操作步骤如下:
步骤一、空白脂质体的制备:
取大豆卵磷脂250g、胆固醇90g和聚氧乙烯脱水山梨醇单油酸酯100g溶解于适量乙醚溶剂中,恒温30-40℃减压旋转蒸发掉有机溶剂,使大豆卵磷脂在蒸发溶器底形成一层薄膜后,加入剩余乙醚溶剂与400ml的磷酸盐缓冲液调节pH7.4,继续旋转蒸发水合20-40分钟后,用0.45微米滤膜过滤后得到空白脂质体溶液,上述乙醚溶剂工100ml;
步骤二、PDRN柔性纳米脂质体的制备:
向空白脂质体溶液中加入PDRN40g、胆酸钠32g,混合均匀后,进行超声3-5分钟,加入200ml的磷酸盐缓冲液,调节pH7.4,继续旋转蒸发水合20-40分钟后,用0.45微米滤膜过滤后得到PDRN柔性纳米脂质体,最后冷处放置保存;
步骤三、采用超速离心法(转速:15000r/min,),离心20min,测定PDRN柔性纳米脂质体的包封率高达96.2%。
对于本领域的技术人员来说,其可以对前述各实施例所记载的技术方案进行修改、部分技术特征的等同替换,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 (4)
1.一种多聚脱氧核糖核苷酸柔性纳米脂质体的制备方法,其特征在于:它的操作步骤如下:
步骤(一)、空白脂质体的制备:
取大豆卵磷脂、胆固醇和聚氧乙烯脱水山梨醇单油酸酯溶解于乙醚溶剂中,其中,大豆卵磷脂重量配比为150-250份,胆固醇重量配比为30-90份,聚氧乙烯脱水山梨醇单油酸酯重量配比为20-100份,恒温30-40℃减压旋转蒸发掉有机溶剂,使大豆卵磷脂在蒸发溶器底形成一层薄膜后,加入乙醚溶剂与磷酸盐缓冲液体积比从1∶1到1∶5,调节pH5-8,继续旋转蒸发水合20-40分钟后,用0.45微米滤膜过滤后得到空白脂质体溶液;
步骤(二)、PDRN柔性纳米脂质体的制备:
向空白脂质体溶液中加入PDRN重量配比为1-40份、胆酸钠20-40份,混合均匀后,进行超声3-5分钟,加入适量的磷酸盐缓冲液,调节pH5-8,继续旋转蒸发水合20-40分钟后,用0.45微米滤膜过滤后得到PDRN柔性纳米脂质体,最后冷处放置保存。
2.根据权利要求1所述的一种多聚脱氧核糖核苷酸柔性纳米脂质体的制备方法,其特征在于:所述PDRN柔性纳米脂质体为球形单室脂质体,形态均一,包封率高达95%以上,粒子直径平均分布在150nm以下。
3.根据权利要求1所述的一种多聚脱氧核糖核苷酸柔性纳米脂质体的制备方法,其特征在于:所述PDRN柔性纳米脂质体中含有PDRN重量配比为5-40份。
4.根据权利要求1所述的一种多聚脱氧核糖核苷酸柔性纳米脂质体的制备方法,其特征在于:所述磷酸盐缓冲液调节pH值为7.4。
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