目的 将聚集引起的猝灭（ACQ）探针P2杂化到槲皮素（QUE）纳米晶的晶格内制备荧光杂化槲皮素纳米晶（QUEHNC），考察QUE-HNC的体外溶出行为及体内药动学。方法 通过沉淀-反溶剂超声法制备QUE-HNC，单因素分析考察滴加速度、探头浸入液面的位置、超声功率、聚维酮K30（PVP K30）的加入方式，筛选制备工艺。制备170、250、310 nm 3种粒径的QUE-HNC，HPLC法检测QUE含量，Nano ZS90激光粒度仪检测粒径及聚合物分散性指数（PDI），Zeta电位分析仪检测电位；扫描电镜观察微观结构；透析袋法比较QUE原料药及不同粒径QUE-HNC的体外溶出行为；雄性昆明小鼠尾iv给予3种粒径的QUE-HNC（126 mg·kg^-1），猝灭组尾iv给予P2水猝灭溶液，给药后0.5、1.0、2.0、4.0、8.0、11.0、24.0 h放入小动物活体三维多模式成像系统检测QUE-HNC的体内组织分布；Wistar大鼠尾iv给予3种粒径的QUE-HNC（100 mg·kg^-1），在0.083、0.250、0.500、1.000、2.000、4.000、8.000、11.000、24.000 h时眼内眦取血0.5 mL进行荧光定量，进行QUE-HNC的粒子动力学研究；HPLC法检测QUE-HNC（100 mg·kg^-1）在大鼠体内的药动学参数。结果 根据单因素考察的结果，最终的处方为：有机相与水的比例为1∶10，PVP K30添加在水相中，有机相的滴加速度为0.01 mL·s^-1。通过调整超声功率和粉碎时间，成功制备170、250、310 nm 3种粒径的QUE-HNC，粒径在所需范围内，PDI均小于0.3，微观形态呈棒状，大小较为均一，QUE质量浓度均在0.40 mg·mL^-1左右；与原料药相比，溶出速度和程度明显提高。3种粒径QUE-HNC的体内分布行为基本一致，在前4 h主要蓄积于肝脏部位，11 h后的荧光信号逐渐向腹部的下方和脾脏处转移，并且随着时间的延长，肝脏部位荧光信号及腹部总荧光信号强度均呈现逐渐减小的趋势。药动学实验的结果反映了粒子快速消除的现象，但粒子动力学结果显示QUE-HNC并不会在血液中快速的溶解，部分QUE-HNC一直处于粒子状态，3种粒径的QUE-HNC在体内都有较长的平均滞留时间。结论 成功制备了3种粒径的QUE-HNC，与槲皮素原料药对比，显著提高了溶出度，但其依然有缓释的效果。

Objective Fluorescently labeled quercetin-hydroxyapatite nanocrystals (QUE-HNC) was prepared by incorporating the aggregation-induced quenching (ACQ) probe P2 into the crystal lattice of quercetin nanocrystals, and investigated the in vitro dissolution behavior and in vivo pharmacokinetics of QUE-HNC. Methods The QUE-HNC was prepared by precipitation-antisolvent ultrasonic method, and single factor analysis was used to investigate the effects of dropping speed, probe immersion position, ultrasonic power, and PVP K30 addition method on the preparation process. The preparation process was optimized to produce QUEHNC with three different sizes of 170, 250, and 310 nm. The content of QUE was determined by HPLC, the size and polymer dispersion index (PDI) were determined by Nano ZS90 laser particle size analyzer, the Zeta potential was determined by Zeta potential analyzer, the microstructure was observed by scanning electron microscopy, the in vitro dissolution behavior of QUE raw material and different sizes of QUE-HNC was compared using dialysis bags, and the in vivo distribution behavior and particle kinetics of QUE-HNC with three different sizes were investigated in male Kunming mice by iv injection of 126 mg·kg^-1 and in Wister rats by iv injection of 100 mg·kg^-1. The pharmacokinetics of QUE-HNC with three different sizes were investigated in Wister rats by iv injection of 100 mg·kg^-1. Results As a result of the single factor investigation, the final formulation was determined to be: A 1:10 ratio of organic phase to water, with PVP K30 added to the aqueous phase, and a drop rate of 0.01 mL·s^-1 for the organic phase. By adjusting the ultrasonic power and grinding time, we successfully prepared QUE-HNC particles with three different sizes of 170, 250, and 310 nm, within the desired range, with PDI all less than 0.3, microscopic morphology in the form of rods, relatively uniform size, and QUE concentration all around 0.40 mg·mL^-1. Compared with the raw drug, the dissolution rate and extent were significantly improved. The in vivo distribution behavior of the three sizes of QUE-HNC was basically the same, with the fluorescence signal mainly accumulating in the liver area in the first 4 h, gradually shifting to the lower abdomen and spleen area after 11 h, and the fluorescence signal intensity in the liver area and the total fluorescence signal intensity gradually decreasing over time. The pharmacokinetic experiment results reflected the phenomenon of rapid elimination of particles, but the particle dynamics results showed that QUE-HNC would not dissolve rapidly in the blood, with some QUE-HNC remaining in particle form. All three sizes of QUE-HNC had a relatively long average residence time in the body. Conclusion Fluorescent hybridized nanocrystals with three particle sizes were successfully prepared, and the nanocrystals significantly improved the dissolution compared to quercetin API, but they still had a slow release effect.

R943

河北省教育厅引进留学人才项目（C20220345）