目的 將聚集引起的猝滅（ACQ）探針P2雜化到槲皮素（QUE）納米晶的晶格內(nèi)制備熒光雜化槲皮素納米晶（QUEHNC），考察QUE-HNC的體外溶出行為及體內(nèi)藥動學。方法 通過沉淀-反溶劑超聲法制備QUE-HNC，單因素分析考察滴加速度、探頭浸入液面的位置、超聲功率、聚維酮K30（PVP K30）的加入方式，篩選制備工藝。制備170、250、310 nm 3種粒徑的QUE-HNC，HPLC法檢測QUE含量，Nano ZS90激光粒度儀檢測粒徑及聚合物分散性指數(shù)（PDI），Zeta電位分析儀檢測電位；掃描電鏡觀察微觀結(jié)構(gòu)；透析袋法比較QUE原料藥及不同粒徑QUE-HNC的體外溶出行為；雄性昆明小鼠尾iv給予3種粒徑的QUE-HNC（126 mg·kg^-1），猝滅組尾iv給予P2水猝滅溶液，給藥后0.5、1.0、2.0、4.0、8.0、11.0、24.0 h放入小動物活體三維多模式成像系統(tǒng)檢測QUE-HNC的體內(nèi)組織分布；Wistar大鼠尾iv給予3種粒徑的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時眼內(nèi)眥取血0.5 mL進行熒光定量，進行QUE-HNC的粒子動力學研究；HPLC法檢測QUE-HNC（100 mg·kg^-1）在大鼠體內(nèi)的藥動學參數(shù)。結(jié)果 根據(jù)單因素考察的結(jié)果，最終的處方為：有機相與水的比例為1∶10，PVP K30添加在水相中，有機相的滴加速度為0.01 mL·s^-1。通過調(diào)整超聲功率和粉碎時間，成功制備170、250、310 nm 3種粒徑的QUE-HNC，粒徑在所需范圍內(nèi)，PDI均小于0.3，微觀形態(tài)呈棒狀，大小較為均一，QUE質(zhì)量濃度均在0.40 mg·mL^-1左右；與原料藥相比，溶出速度和程度明顯提高。3種粒徑QUE-HNC的體內(nèi)分布行為基本一致，在前4 h主要蓄積于肝臟部位，11 h后的熒光信號逐漸向腹部的下方和脾臟處轉(zhuǎn)移，并且隨著時間的延長，肝臟部位熒光信號及腹部總熒光信號強度均呈現(xiàn)逐漸減小的趨勢。藥動學實驗的結(jié)果反映了粒子快速消除的現(xiàn)象，但粒子動力學結(jié)果顯示QUE-HNC并不會在血液中快速的溶解，部分QUE-HNC一直處于粒子狀態(tài)，3種粒徑的QUE-HNC在體內(nèi)都有較長的平均滯留時間。結(jié)論 成功制備了3種粒徑的QUE-HNC，與槲皮素原料藥對比，顯著提高了溶出度，但其依然有緩釋的效果。

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

河北省教育廳引進留學人才項目（C20220345）