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    • 5. 发明授权
    • Composition containing small multilamellar oligodeoxynucleotide-containing lipid vesicles
    • 含有小的多层寡脱氧核苷酸的脂质囊泡的组合物
    • US06835395B1
    • 2004-12-28
    • US09654373
    • 2000-09-01
    • Sean C. SempleSandra K. KlimukTroy O. HarasymNancy Dos SantosSteven M. AnsellPieter R CullisMichael J. HopePeter ScherrerDeirdre McIntoshKim F. WongNorbert Maurer
    • Sean C. SempleSandra K. KlimukTroy O. HarasymNancy Dos SantosSteven M. AnsellPieter R CullisMichael J. HopePeter ScherrerDeirdre McIntoshKim F. WongNorbert Maurer
    • A61K9127
    • A61K9/1272Y10T428/2984
    • Lipidic compositions with superior characteristics for in vivo delivery of oligodeoxynucleotides (ODN) can easily and efficiently be made in the form of small multilamellar vesicles. The compositions contain a population of nucleic acid-containing lipid vesicles in a liquid carrier, and at least a portion of the lipid vesicles are small multilamellar vesicles. The small multilamellar vesicles are made from a lipid component including 20-30 mol % of an ionizable amino lipid such as DODAP, and a steric barrier lipid such as PEG-CerC14; and an oligodeoxynucleotide contained in the lumen or interlamellar spaces of the small multilamellar vesicles. The ODN and lipid components are preferably present in the small multilamellar vesicles in a mole ratio of from 0.15 to 0.25. The compositions of the invention can be made by preparing two solutions: a lipid mixture with 20-30 mol % of the ionizable amino lipid, the steric barrier lipid and additional lipid components selected from among neutral lipids and sterols in an ethanolic solvent; and a solution of oligodeoxynucleotide in an aqueous solvent having a pH at which the ionizable amino lipid is positively charged. The lipid mixture is added to the solution of oligodeoxynucleotide to form a mixture containing lipid vesicles which is then massed through a filter to produce sized lipid vesicles in a solution containing ethanol. The ethanol is then removed, for example by dialysis. Then, the pH of the solution surrounding the sized lipid vesicles is increased to reduce the net positive charge on the exterior of the sized lipid vesicles.
    • 寡脱氧核苷酸(ODN)的体内递送具有优异特性的脂质体组合物可以容易且有效地以小的多层囊泡的形式进行。 组合物在液体载体中含有含有核酸的脂质囊泡,并且至少一部分脂质囊泡是小的多层囊泡。 小的多层泡囊由包含20-30mol%的可离子化氨基脂质如DODAP和空间屏障脂质如PEG-CerC14的脂质组分制成; 和包含在小多层囊泡的内腔或层间空间中的寡脱氧核苷酸。 ODN和脂质组分优选以0.15至0.25的摩尔比存在于小多层囊泡中。 本发明的组合物可以通过制备两种溶液:在乙醇溶剂中具有20-30mol%的可电离氨基脂质的脂质混合物,空间屏障脂质和选自中性脂质和甾醇中的另外的脂质组分; 和寡脱氧核苷酸在具有可电离氨基脂质带正电荷的pH的水性溶剂中的溶液。 将脂质混合物加入到寡脱氧核苷酸的溶液中以形成含有脂质囊泡的混合物,然后通过过滤器将其混合,以在含有乙醇的溶液中产生大小的脂质囊泡。 然后例如通过透析除去乙醇。 然后,增加围绕大小的脂质囊泡的溶液的pH以降低大小的脂质囊泡外部的净正电荷。