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    • 5. 发明公开
    • 단백질을 포위하는 나노리포좀의 제조방법 및 단백질-포위나노리포좀
    • 制备纳米脂肪酸包封蛋白和蛋白质包被的纳米脂质体的方法
    • KR1020080000602A
    • 2008-01-02
    • KR1020077024282
    • 2006-03-28
    • 주식회사 리제론
    • 오달균이균영
    • A61K49/18A61K49/06A61P43/00
    • A61K9/0014A61K8/14A61K8/553A61K8/64A61K9/127A61K9/1277A61K2800/413A61Q5/12A61Q7/00A61Q19/00A61Q19/08B82Y5/00
    • A method for preparing nanoliposomes is provided to obtain the nanoliposomes which have high encapsulation efficiency for proteins, are in the form of liquid, have a desired particle size and maintain their original activity by applying the stepwisely increased shearing force to a composition capable of forming the nanoliposomes with a stepwise addition of phospholipids. A method for preparing a nanoliposome encapsulating a protein comprises the steps of: (a) dispersing a phospholipid in an aqueous solution containing a protein to prepare a dispersion; (b) applying a shearing force to the dispersion; (c) adding an additional amount of the phospholipid to the resultant of the step(b) and then applying a shearing force higher than that of the step(b); and (d) repeating the step(c) with an additional amount of the phospholipid and a shearing force higher than a prior step to obtain the nanoliposome having a diameter and an desired encapsulation efficiency, wherein the protein is selected from the group consisting of hormone, hormone analogue, enzyme, enzyme inhibitor, signal transduction protein or fragment thereof, antibody or fragment thereof, single chain antibody, binding protein or fragment thereof, peptide, antigen, adhesive protein, structural protein, regulatory protein, toxin protein, cytokine, transcription regulatory protein, blood clotting protein and plant defense-inducing protein, the phospholipid is at least one selected from the group consisting of soybean lecithin, distearoylphosphatidylcholine, hydrogenated soybean lecithin, egg lecithin, dioleoylphosphatidylcholine, hydrogenated egg lecithin, dielaidoylphosphatidylcholine, dipalmitoylphosphatidylcholine and dimyristoylphosphatidylcholine, the application of the shearing force is performed using a high pressure homogenizer, a sonicator, a microfluidizer, an extrusion apparatus or a French press. A protein-encapsulating nanoliposome is characterized in that the nanoliposome has the diameter of 50-350 nm, has the structure of a small unilamellar vesicle, and is in the form of liquid, and the protein encapsulated in the nanoliposome has an activity corresponding to 90-100% of that of the protein prior to encapsulation.
    • 提供了一种制备纳米脂质体的方法,以获得对于蛋白质具有高包封效率的纳米脂质体,其为液体形式,具有期望的粒度并通过将逐步增加的剪切力施加到能够形成 纳米脂质体逐步加入磷脂。 制备包封蛋白质的纳米脂质体的方法包括以下步骤:(a)将磷脂分散在含有蛋白质的水溶液中以制备分散体; (b)对分散体施加剪切力; (c)向步骤(b)的所得物中加入额外量的磷脂,然后施加高于步骤(b)的剪切力; 和(d)用额外量的磷脂和高于先前步骤的剪切力重复步骤(c),以获得具有直径和期望的包封效率的纳米脂质体,其中所述蛋白质选自激素 ,激素类似物,酶,酶抑制剂,信号转导蛋白或其片段,抗体或其片段,单链抗体,结合蛋白或其片段,肽,抗原,粘附蛋白,结构蛋白,调节蛋白,毒素蛋白,细胞因子,转录 调节蛋白,凝血蛋白和植物防御诱导蛋白,磷脂是选自大豆卵磷脂,二硬脂酰磷脂酰胆碱,氢化大豆卵磷脂,卵磷脂,二油酰磷脂酰胆碱,氢化蛋卵磷脂,二酰基磷脂酰胆碱,二棕榈酰磷脂酰胆碱和二肉豆蔻酰磷脂酰胆碱中的至少一种, 她的应用 使用高压均化器,超声波发生器,微流化器,挤出装置或法式压力机进行施加力。 蛋白质包封的纳米脂质体的特征在于纳米脂质体具有50-350nm的直径,具有小的单层囊泡的结构,并且是液体形式,并且包封在纳米脂质体中的蛋白质具有对应于90 -100%的蛋白质在封装之前。