CAJ | 학술논문

目的：考察银杏内酯 B 前体药物（PGB）的脑靶向性，并探究其靶向机制。方法采用 LC-MS ／MS 考察大鼠尾静脉注射 PGB 后的脑部药动学规律并评价其脑靶向性；采用伊文斯蓝法观察 PGB 对不完全性脑缺血小鼠脑毛细血管通透性的影响；HPLC 法测定 PGB 在正辛醇-水体系的分配系数（logP）；MVD 分子对接软件计算 PGB 与 P-糖蛋白（P-gp）的体外结合力；定磷法检测 PGB 对人 P-gp 膜 ATP 酶活性的影响。结果以治疗有效性和靶向指数评价 PGB 的脑靶向效率达6.87和4.14；PGB 预防给药可有效降低不完全性脑缺血小鼠的脑毛细血管通透性（P ＜0.05）；PGB 的 logP 为1.03，高于 GB 的0.61；分子对接计算表明 PGB 与 P-gp 结合力大于 GB，其 MolDockScore 分别为－143.36、－116.40 KJ ·mol －1；ATP 酶活性分析提示 PGB、GB 均能提高 P-gp ATP酶活性，其 Km 值分别为237.75、841.24μmol·L －1，PGB 与P-gp 亲和力高于 GB。结论PGB 具有脑靶向性，其靶向性提高一方面得益于其脂溶性提高，PGB 在脑部渗透量增加；另一方面可能为 PGB 明显提高 P-gp ATP 酶活性，从而抑制P-gp 对 GB 的外排作用。
목적：고찰은행내지 B 전체약물（PGB）적뇌파향성，병탐구기파향궤제。방법채용 LC-MS ／MS 고찰대서미정맥주사 PGB 후적뇌부약동학규률병평개기뇌파향성；채용이문사람법관찰 PGB 대불완전성뇌결혈소서뇌모세혈관통투성적영향；HPLC 법측정 PGB 재정신순-수체계적분배계수（logP）；MVD 분자대접연건계산 PGB 여 P-당단백（P-gp）적체외결합력；정린법검측 PGB 대인 P-gp 막 ATP 매활성적영향。결과이치료유효성화파향지수평개 PGB 적뇌파향효솔체6.87화4.14；PGB 예방급약가유효강저불완전성뇌결혈소서적뇌모세혈관통투성（P ＜0.05）；PGB 적 logP 위1.03，고우 GB 적0.61；분자대접계산표명 PGB 여 P-gp 결합력대우 GB，기 MolDockScore 분별위－143.36、－116.40 KJ ·mol －1；ATP 매활성분석제시 PGB、GB 균능제고 P-gp ATP매활성，기 Km 치분별위237.75、841.24μmol·L －1，PGB 여P-gp 친화력고우 GB。결론PGB 구유뇌파향성，기파향성제고일방면득익우기지용성제고，PGB 재뇌부삼투량증가；령일방면가능위 PGB 명현제고 P-gp ATP 매활성，종이억제P-gp 대 GB 적외배작용。
Aim To investigate the brain targeting of ginkgolide B prodrug (PGB ) and its mechanism. Methods The liquid chromatography tandem mass spectrometry (LC-MS /MS)method was applied to in-vestigate the pharmacokinetics of PGB in rat brain tis-sue after intravenous injection of PGB.Also the brain targeting was evaluated on the basis of the pharmacoki-netic parameter of PGB.The incomplete cerebral is-chemia model was induced in mouse,the effect of PGB on cerebral capillary permeability was observed by Ev-ans blue method.High performance liquid chromatog-raphy (HPLC)was used to determine the partition co-efficients (logP)of PGB in octanol-water system.PGB and P-glycoprotein (P-gp)was docked by using Mole-gro Virtual Docker (MVD)software to predict its bind-ing abilities with P-gp.The interaction of PGB with ATPase activity of human P-gp membrane was esti-mated by measuring inorganic phosphate liberation. Results The brain targeting of PGB was evaluated by treatment effective (TA ) and drug targeting index (DTI),the calculated value were 6.87 and 4.1 4 re-spectively.Preventive medication of PGB could signifi-cantly decrease cerebral capillary permeability (P <0.05).The lipo-hydropartition coefficient of PGB was higher than that of GB,their logP data were 1 .03 and 0.61 respectively.PGB displayed the stronger binding affinity with P-gp than GB according to the molecular docking calculations,their MolDock Score toward P-gp were -1 43.36 and -1 1 6.40KJ·mol -1 respectively. ATP-hydrolisis showed that PGB increased ATPase activity with a Km of approximately 237.75 μmol · L -1 ,however GB with a Km of approximately 841 .24μmol·L -1 .PGB might interact with P-gp with a high-er affinity and exhibit more effect than GB.Conclu-sion PGB is characterized by its brain targeting. Higher liposolubility of PGB results in good blood-brain permeability,which is advantageous to its brain targe-ting.Besides,PGB can effectively inhibit the efflux effect of P-gp to GB because of its increased P-gp AT-Pase activity.