目的 基于頂端鈉依賴性膽鹽轉(zhuǎn)運體（ASBT）對膽酸（BA）的特異性識別，為提高阿苯達唑（ABZ）的跨膜轉(zhuǎn)運效率，制備阿苯達唑-膽酸衍生物（ABZ-C6O2-BA），并對其固態(tài)性質(zhì)、跨膜轉(zhuǎn)運行為和降解機制進行評價。方法 以6-氨基-1-羥基-二己脂作為Linker連接ABZ與BA，合成ABZ-C6O2-BA；使用核磁共振氫譜（¹H-NMR）和傅里葉紅外技術(shù)（FTIR）對ABZ-C6O2-BA進行驗證；通過粉末X射線衍射（PXRD）法、差示掃描量熱（DSC）法和掃描電子顯微鏡（SEM）法進行固態(tài)性質(zhì)的表征；建立Caco-2單層細胞模型在體外對ABZ-C6O2-BA進行跨膜轉(zhuǎn)運研究；進行ABZ-C6O2-BA體外羧酸酯酶（CES）水解反應，通過超高效液相色譜-質(zhì)譜聯(lián)用（UPLC-MS）定性分析ABZ-C6O2-BA水解產(chǎn)物。結(jié)果 ¹H-NMR和FT-IR結(jié)果顯示特征峰的偏移，證實ABZ-C6O2-BA制備成功；PXRD、DSC和SEM結(jié)果顯示ABZ-C6O2-BA為無定型形態(tài)，且微觀結(jié)構(gòu)較ABZ有明顯不同；體外跨膜轉(zhuǎn)運實驗結(jié)果顯示高質(zhì)量濃度的ABZ-C6O2-BA跨膜轉(zhuǎn)運表觀滲透系數(shù)（P_app）顯著高于高濃度的ABZ（P＜0.05），中、低質(zhì)量濃度的ABZ-C6O2-BA P_app相近于ABZ；ABZ-C6O2-BA在72 h時降解率＞92.8%，代謝良好；通過UPLC-MS定性分析ABZ-C6O2-BA水解產(chǎn)物為ABZ-Linker，即CES水解了酯鍵，使得BA與ABZ分開。結(jié)論 所制備的ABZ-C6O2-BA以無定型態(tài)存在，跨膜轉(zhuǎn)運效率在一定濃度下較ABZ顯著增加，且與濃度不相關(guān)，推測跨膜方式為主動轉(zhuǎn)運，連接的BA可被體內(nèi)的酶水解。

Objective To improve the transcellular transport efficiency of albendazole (ABZ) by preparing a novel compound, ABZC6O2-BA, based on the specific recognition of the apical sodium-dependent bile acid transporter (ASBT) for bile acid (BA). The solid state properties, transcellular transport behavior, and degradation mechanism of ABZ-C6O2-BA were evaluated. Method ABZC6O2-BA was synthesized by linking ABZ with BA using 6-aminodihydroxy-dipentyl glycerol as the linker. The synthesized ABZC6O2-BA was verified by ¹H-NMR and FT-IR. The solid state properties were characterized by powder X-ray diffraction (PXRD), differential scanning calorimetry (DSC), and scanning electron microscopy (SEM). The transcellular transport behavior of ABZC6O2-BA was studied in vitro using a Caco-2 monolayer cell model. The degradation mechanism of ABZ-C6O2-BA was evaluated by in vitro carboxylesterase (CES) hydrolysis, and the hydrolysis products were quantitatively analyzed by ultra-high-performance liquid chromatography-mass spectrometry (UPLC-MS). Results The ¹H-NMR and FT-IR results showed a shift in characteristic peaks, confirming the successful synthesis of ABZ-C6O2-BA. The PXRD, DSC, and SEM results showed that ABZ-C6O2-BA was amorphous in form, and its microstructure was significantly different from that of ABZ. The in vitro transcellular transport experiment results showed that the apparent permeability coefficient (P_app) of high-quality ABZ-C6O2-BA across the membrane was significantly higher than that of high-concentration ABZ (P < 0.05), and the P_app of medium-and low-quality ABZ-C6O2-BA was similar to that of ABZ. The degradation rate of ABZ-C6O2-BA was > 92.8% at 72 h. By UPLC-MS qualitative analysis, the hydrolysis product of ABZ-C6O2-BA was identified as ABZ-Linker, indicating that the CES hydrolyzed the ester bond, separating BA from ABZ. Conclusion The ABZ-C6O2-BA prepared in this study exists in an amorphous state, and its transmembrane transport efficiency is significantly increased at certain concentrations, which is independent of the concentration, suggesting that the transmembrane mode is active transport. The connected BA can be hydrolyzed by enzymes in the body.

R943

2022年青海省科技廳科技援青合作專項（2022-QY-201）