麦角甾醇联合阿法替尼双载药脂质体的制备及质量评价

Preparation and Quality Evaluation of Ergosterol and Afatinib Double-loaded Liposomes

目的研究麦角甾醇(ERG)联合阿法替尼(Afa)双载药脂质体的最佳制备工艺,并对其进行质量评价。方法以薄膜分散法制备ERG脂质体(ERG-LIP),用硫酸铵梯度法将Afa柠檬酸溶液载入脂质体内水相,制备ERG联合Afa双载药脂质体(ERG/Afa-LIP),以Sephadex-G50微柱离心法测定ERG/Afa-LIP的包封率。以单因素试验结果为基础,用Box-Behnken试验考察各因素对脂质体包封率的影响,确定ERG/Afa-LIP的最佳制备工艺。对该工艺下的ERG/Afa-LIP进行形貌表征,测定其粒径与Zeta电位、pH值、过氧化值、包封率与载药量、体外释放度。结果确定ERG/Afa-LIP最佳制备工艺为孵育温度54.2℃,孵育时间59.06 min,Afa浓度0.238 mg·mL^(-1),硫酸铵浓度194.63 mmol·L-1。外观形态较完整,平均粒径为(103.03±0.61)nm,聚合物分散性指数(PDI)为(0.261±0.003),Zeta电位为(2.48±0.45)mV,pH均值为(3.78±0.02),显酸性,平均包封率为(96.92±0.58)%,载药量为(2.92±0.23)%,在释放介质为pH6.5的含40%甲醇的磷酸盐缓冲液下,ERG/AfaLIP的体外释放度符合《中国药典》要求。结论制备出具有靶向作用的ERG与Afa双载药脂质体,其质量稳定,符合2015年版《中国药典》的要求。

Objective To study the best preparation process of ergosterol(ERG)combined with afatinib(Afa)double-loaded liposomes,and to evaluate its quality.MethodsThe ergosterol liposomes(ERG-LIP)were preparedby thin-film dispersion method,and the afatinib citric acid solution was loaded into the liposome aqueous phase bythe ammonium sulfate gradient method to prepare ergosterol combined with afatinib double-loaded drug liposome(ERG/Afa-LIP).Sephadex-G50 microcolumn centrifugation method was used to determine the encapsulationefficiency of ERG/Afa-LIP.Based on the single-factor test results,the effects of various factors on the liposomeencapsulation rate were investigated according to the principle of Box-Behnken test design,and the best preparationprocess of ERG/Afa-LIP was determined.The morphological characterization of ERG/Afa-LIP was performed,andits particle size and Zeta potential,p H value,peroxide value,encapsulation rate and drug loading,in vitro releaserate were determined.ResultsThe best preparation process for ERG/Afa-LIP was determined to be as follows:anincubation temperature of 54.2℃,and on incubation time of 59.06 min,Afa concentration at 0.238 mg·mL^(-1),and ammonium sulfate concentration at 194.63 m M.The appearance is relatively round,and the average particle size is(103.03±0.61)nm,the PDI is(0.261±0.003),the Zeta potential is(2.48±0.45)m V;it is acidic with anaverage p H of(3.78±0.02),and the average encapsulation rate is(96.92±0.58)%;the drug loading is(2.92±0.23)%.The ERG/Afa-LIP meets the requirements of the Chinese Pharmacopoeia in the release medium of p H 6.5 phosphate buffer containing 40%methanol.Conclusion ERG and Afa dual drug-loaded liposomes with targetingeffect was prepared,and the quality was stable,meeting the requirements of the 2015 edition of Chinese Pharmacopoeia.