丹参酮Ⅱ_(A)聚合物脂质纳米粒制备及脑部药物递送研究  

Study on preparation of tanshinone ⅡA polymer lipid nanoparticles and brain delivery

作　　者：高羚毓 贾瑞欣 毕野 邱智东[1] 李银清 苏婷[1] 邱野[1] GAO Lingyu;JIA Ruixin;BI Ye;QIU Zhidong;LI Yinqing;SU Ting;QIU Ye(Collage of Pharmacy,Changchun University of Traditional Chinese Medicine,Changchun 130117,China)

机构地区：[1]长春中医药大学药学院,吉林长春130117

出　　处：《药物评价研究》2022年第5期909-917,共9页Drug Evaluation Research

基　　金：吉林省发展改革委员会项目(202000C032-4)。

摘　　要：目的为了改善丹参酮Ⅱ_(A)(TanⅡ_(A))的溶解性、增加脑组织中的药物浓度,制备TanⅡ_(A)聚合物脂质纳米粒(TanⅡ_(A)-PLNs)并进行体外释放、药动学、脑组织分布研究。方法建立TanⅡ_(A)含量测定的高效液相色谱(HPLC)法,以纳米制剂的包封率和粒径为指标,优化TanⅡ_(A)-PLNs的制备工艺,分别优化TanⅡ_(A)与蛋黄磷脂(Egg-PC)的比例、Egg-PC与聚乳酸-羟基乙酸共聚物(PLGA)的比例、有机相与水相的比例;进行TanⅡ_(A)-PLNs的处方验证、稳定性考察、体外释放考察。建立TanⅡ_(A)在血浆和脑匀浆中的液-质联用(LC-MS)检测方法。SPF级雄性SD大鼠6只,随机分为2组,给药前12 h禁食,一组ig 25 mg·kg^(-1)的TanⅡ_(A)混悬液,另一组尾iv 5 mg·kg^(-1)的TanⅡ_(A)-PLNs溶液,于给药后5、15、30、60、120、240、360、480、720 min眼眶取血约0.5 m L,LC-MS法进行药动学检测。取KM小鼠4只,尾iv Di R标记的Tan Ⅱ_(A)-PLNs,给药30、60、120、240 min后处死解剖,通过荧光成像观察Tan Ⅱ_(A)-PLNs在组织中的分布情况。取KM小鼠12只,随机分成4组,给药前12 h禁食,尾iv 2.5 mg·kg^(-1)的TanⅡ_(A)-PLNs溶液,于给药后30、60、120、240 min处死小鼠,剥离完整大脑组织,称质量,加入2倍超纯水匀浆,LC-MS法检测TanⅡ_(A)水平。结果TanⅡ_(A)-PLNs的最优处方为TanⅡ_(A)∶Egg-PC为1∶4、Egg-PC∶PLGA为5∶2、有机分散体系∶水分散体系为1∶15。以Egg-PC/PLGA作为脂质纳米粒的膜材,采用纳米粒沉淀法制备的TanⅡ_(A)-PLNs的平均粒径为272 nm,Zeta电位为-4.93 m V,包封率为88.2%,载药量为18%。在避光、干燥(室温)的条件下Tan Ⅱ_(A)-PLNs冻干粉稳定。Tan Ⅱ_(A)原料药在48 h内释放完全,累积释放率为(93.75±0.75)%,与Korsmeyer-Peppas释放方程拟合程度较好;Tan Ⅱ_(A)-PLNs在400 h左右释放完全,累积释放率为(89.44±2.22)%,与零级释放方程拟合度最好。药动学结果表明,大鼠尾iv TanⅡ_(A)-PLNs给药剂量是ig Tan�Objective Tanshinone Ⅱ_(A)polymeric lipid nanoparticles(TanⅡ_(A)-PLNs)were prepared for improving the solubility and drug concentration in brain tissue to increase drug brain exposure.In vitro release,pharmacokinetics and brain tissue distribution were studied.Method A high performance liquid chromatography(HPLC)method was established to determine the content of TanⅡ_(A),and the preparation process of TanⅡ_(A)-PLNs was optimized by taking the encapsulation rate and particle size of nano-preparations as indexes.The proportion of TanⅡ_(A)to Egg-PC,Egg-PC to polylactic acid-hydroxyacetic acid copolymer(PLGA)and organic phase to water phase were optimized respectively.The prescription validation,stability and in vitro release of TanⅡ_(A)-PLNs were investigated.To establish a method for the determination of TanⅡ_(A)in plasma and brain homogenates by liquid-mass spectrometry(LC-MS).Six SPF male SD rats were randomly divided into two groups,one group was given 25 mg·kg^(-1) TanⅡ_(A)suspension,the other group was given 5 mg·kg^(-1) TanⅡ_(A)-PLNs solution.At 5,15,30,60,120,240,360,480,720 min after administration,about0.5 m L blood was collected from orbit,and pharmacokinetic detection was performed by LC-MS.Four KM mice were selected and TanⅡ_(A)-PLNs labeled with iv Di R were sacrificed 30,60,120 and 240 min after administration,and the distribution of TanⅡ_(A)-PLNs in tissues was observed by fluorescence imaging.Twelve KM mice were randomly divided into four groups,fasted 12 h before administration,and treated with 2.5 mg·kg^(-1) TanⅡ_(A)-PLNs solution in tail iv.The mice were sacrificed 30,60,120 and 240 min after administration.The intact brain tissues were stripped,weighed,and the level of TanⅡ_(A)was detected by LC-MS.Results Egg-PC/PLGA(5:2)were used as membrane materials of Tan Ⅱ_(A)-PLNs nanoparticles,and the volume ratio of organic phase to water phase was 1:15.The particle size,zeta potential,entrapment efficiency and drug loading capacity of Tan Ⅱ_(A)-PLNs were 272 nm,-4.93 m

关 键 词：丹参酮Ⅱ_(A) 聚合物脂质纳米粒 脑组织分布 药动学 

分 类 号：R943[医药卫生—药剂学] R969.1[医药卫生—药学]