目的 為了改善丹參酮Ⅱ_A（TanⅡ_A）的溶解性、增加腦組織中的藥物濃度，制備TanⅡ_A聚合物脂質(zhì)納米粒（TanⅡ_APLNs）并進(jìn)行體外釋放、藥動(dòng)學(xué)、腦組織分布研究。方法 建立TanⅡ_A含量測(cè)定的高效液相色譜（HPLC）法，以納米制劑的包封率和粒徑為指標(biāo)，優(yōu)化TanⅡ_A-PLNs的制備工藝，分別優(yōu)化TanⅡ_A與蛋黃磷脂（Egg-PC）的比例、Egg-PC與聚乳酸-羥基乙酸共聚物（PLGA）的比例、有機(jī)相與水相的比例；進(jìn)行TanⅡ_A-PLNs的處方驗(yàn)證、穩(wěn)定性考察、體外釋放考察。建立TanⅡ_A在血漿和腦勻漿中的液-質(zhì)聯(lián)用（LC-MS）檢測(cè)方法。SPF級(jí)雄性SD大鼠6只，隨機(jī)分為2組，給藥前12 h禁食，一組ig 25 mg·kg^-1的TanⅡ_A混懸液，另一組尾iv 5 mg·kg^-1的TanⅡ_A-PLNs溶液，于給藥后5、15、30、60、120、240、360、480、720 min眼眶取血約0.5 mL，LC-MS法進(jìn)行藥動(dòng)學(xué)檢測(cè)。取KM小鼠4只，尾iv DiR標(biāo)記的TanIIA-PLNs，給藥30、60、120、240 min后處死解剖，通過熒光成像觀察TanIIA-PLNs在組織中的分布情況。取KM小鼠12只，隨機(jī)分成4組，給藥前12 h禁食，尾iv 2.5 mg·kg^-1的TanⅡ_A-PLNs溶液，于給藥后30、60、120、240 min處死小鼠，剝離完整大腦組織，稱質(zhì)量，加入2倍超純水勻漿，LC-MS法檢測(cè)TanⅡ_A水平。結(jié)果 TanⅡ_A-PLNs的最優(yōu)處方為TanⅡ_A∶Egg-PC為1∶4、Egg-PC∶PLGA為5∶2、有機(jī)分散體系∶水分散體系為1∶15。以Egg-PC/PLGA作為脂質(zhì)納米粒的膜材，采用納米粒沉淀法制備的TanⅡ_A-PLNs的平均粒徑為272 nm，Zeta電位為-4.93 mV，包封率為88.2%，載藥量為18%。在避光、干燥（室溫）的條件下TanIIA-PLNs凍干粉穩(wěn)定。TanIIA原料藥在48 h內(nèi)釋放完全，累積釋放率為（93.75±0.75）%，與Korsmeyer-Peppas釋放方程擬合程度較好； TanIIAPLNs在400 h左右釋放完全，累積釋放率為（89.44±2.22）%，與零級(jí)釋放方程擬合度最好。藥動(dòng)學(xué)結(jié)果表明，大鼠尾iv TanⅡ_A-PLNs給藥劑量是ig TanⅡ_A原料藥劑量的1/5，TanⅡ_A-PLNs的AUC_0-t和t_1/2ß是TanⅡ_A原料藥的2.8和1.5倍。組織分布實(shí)驗(yàn)結(jié)果顯示，30 min時(shí)肝組織顯示熒光； 60 min時(shí)腦組織顯示出熒光，120 min時(shí)腦組織熒光最強(qiáng)，240 min時(shí)腦組織熒光變?nèi)?。TanⅡ_A-PLNs給藥2 h后，TanⅡ_A腦勻漿質(zhì)量濃度為235.5 ng·g^-1。結(jié)論 制備的TanⅡ_A-PLNs均一穩(wěn)定，包封率較高，聚合物脂質(zhì)納米顆粒的應(yīng)用提高了TanⅡ_A的血藥濃度，可增加藥物在腦部的暴露。

Objective Tanshinone IIA 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 Tan Ⅱ_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, about 0.5 mL 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 DiR 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 TanIIA-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 TanIIA-PLNs were 272 nm, - 4.93 mV, 88.2%, 18% respectively. TanⅡ_A-PLNs freeze-dried powder is stable under dry conditions (room temperature) and away from light. Pharmacokinetics showed that the AUC _(0-t) and t_1/2ß of TanIIA-PLNs were 2.8 and 1.5 times higher than free TanIIA even the dose difference of administration was 1/5 times. The results of tissue distribution experiment showed that the liver tissue showed fluorescence at 30 min. The brain tissue showed fluorescence at 60 min, the fluorescence was the strongest at 120 min, and the fluorescence became weak at 240 min. The plasma concentration of TanIIA reached 235.5 ng/g after 2 h of TanIIA-PLNs administration. Conclusion TanIIA-PLNs were uniform and stable, and had a higher entrapment efficiency. In vivo pharmacokinetic parameters showed that the application of polymeric lipid nanoparticles improved the concentration of TanIIA in brain tissue and increased the exposure of the drug in the brain.

R943;R969.1

吉林省發(fā)展改革委員會(huì)項(xiàng)目(202000C032-4)