目的 优化黄芩汤自组装纳米粒（self-assembled nanoparticles of Huangqin Decoction，HQD-SAN）与特比萘芬（terbinafine，TBF）共载药纳米粒（TBF-HQD-SAN NPs）的处方工艺。方法 采用高速离心结合透析法拆分得到HQD-SAN，进一步包载TBF制备为TBF-HQD-SAN NPs。在单因素考察的基础上，以HQD-SAN质量浓度、磁力搅拌速度和搅拌时间为考察因素，以TBF载药量、黄芩苷包封率及载药量为考察指标，采用3因素3水平Box-Behnken设计（Box-Behnken design，BBD）-效应面法（response surface methodology，RSM）优化TBF-HQD-SAN NPs的处方和工艺。对最优处方和工艺制备的TBF-HQD-SAN NPs进行形貌、粒径分布、ζ电位及载药能力、溶解度进行表征，并考察其对红色毛癣菌Trichophyton rubrum、须癣毛癣菌T. mentagrophytes、犬小孢子菌Microsporum canis的抗菌活性。结果 工艺优化分析表明，所建2次回归模型拟合度优异（R²均＞0.99），HQD-SAN质量浓度、搅拌速度、搅拌时间及部分交互项对指标影响显著（P＜0.05）。效应面分析显示，TBF载药量随HQD-SAN质量浓度升高而降低，随磁力搅拌速度、磁力搅拌时间先增后降；黄芩苷包封率、载药量随HQD-SAN质量浓度升高而增加，随磁力搅拌速度、磁力搅拌时间延长而降低。模型优选最优工艺为HQD-SAN质量浓度5.6 mg/mL，TBF 5 mg，蒸馏水4 mL，超声（50 W、40 kHz）30 min，760 r/min磁力搅拌1.5 h，0.8 μm滤膜滤过，即得；验证实验中各指标实测值与预测值接近（RSD＜5%）。所得TBF-HQD-SAN NPs的粒径为（185.10±1.73）nm，多分散指数（polydispersity index，PDI）为0.22±0.01，ζ电位为（−15.17±1.40）mV；TBF包封率为（99.81±0.33）%，载药量为（3.32±0.09）%；黄芩苷包封率为（58.59±1.42）%、载药量为（6.71±0.15）%；TBF、TBF-HQD-SAN物理混合物（physical mixture，PM）、TBF-HQD-SAN NPs的平衡溶解度分别为（2.27±0.09）、（15.70±1.66）、（78.20±2.22）μg/mL。抗菌实验结果显示，TBF-HQD-SAN NPs的抗菌活性（MIC值为0.15～0.31µg/mL，以TBF计为4.98～10.13 ng/mL）显著优于HQD-SAN（MIC值为1.56～3.13 mg/mL）、TBF（MIC值为0.06～0.50µg/mL）（P＜0.05）。结论 BBD成功优化了TBF-HQD-SAN NPs处方和工艺，该制剂粒径均一、载药性能优异，且抗菌效果显著提升，为其后续研究奠定了基础。

Objective To optimize the formulation process of co-loaded nanoparticles (TBF-HQD-SAN NPs) comprising self-assembled nanoparticles of Huangqin Decoction (黄芩汤) (HQD-SAN) and terbinafine (TBF). Methods The HQD-SAN was obtained with high-speed centrifugation combined with dialysis and then TBF was further loaded to prepare TBF-HQD-SAN NPs. Based on the single-factor investigation, taking the concentration of HQD-SAN, the speed of magnetic stirring and the stirring time as the investigation factors, and the drug loading (DL) of TBF, the encapsulation efficiency and the DL of baicalin as the investigation indicators, the formulation and process of TBF-HQD-SAN NPs were optimized by using the 3-factor 3-level Box-Behnken design (BBD)-response surface methodology (RSM). The optimized TBF-HQD-SAN NPs were characterized for morphology, particle size distribution, ζ potential, and drug-loading capacity. Their saturated solubility was determined, and their antifungal activity against Trichophyton rubrum, T. mentagrophytes, and Microsporum canis was investigated. Results The TBF-HQD-SAN NPs process was optimized by BBD. The established quadratic regression model had excellent fit (all R² > 0.99), and the concentration of HQD-SAN, stirring speed, stirring time and some interaction terms had significant effects on the indicators (P < 0.05). The effect surface analysis showed that the drug loading of TBF decreased with the increase of HQD-SAN concentration and first increased and then decreased with the stirring speed/time. The encapsulation rate/drug loading of baicalin increases with the increased of HQD-SAN concentration and decreased with the extension of stirring speed/time. The optimal process for model selection was as follows: HQD-SAN concentration 5.6 mg/mL, TBF 5 mg, distilled water 4 mL, ultrasonic (50 W, 40 kHz) for 30 min, magnetic stirring at 760 r/min for 1.5 h, and filtration through 0.8 μm filter membrane. The measured values of each index in the verification experiment were close to the predicted values (RSD < 5%). The particle size of the NPs was (185.10 ± 1.73) nm, the polydispersity index (PDI) was 0.22 ± 0.01, and the ζ potential was (−15.17 ± 1.40) mV. The encapsulation efficiency and DL of TBF were (99.81 ± 0.33)% and (3.32 ± 0.09)%, respectively, while those of baicalin were (58.59 ± 1.42)% and (6.71 ± 0.15)%, respectively. The equilibrium solubility was (2.27 ± 0.09) μg/mL for TBF alone, (15.70 ± 1.66) μg/mL for the TBF-HQD-SAN physical mixture (PM), and (78.20 ± 2.22) μg/mL for the TBF-HQD-SAN NPs. The antifungal experiment showed that the antifungal activity of TBF-HQD-SAN NPs (with MIC values ranging from 0.15—0.31 µg/mL, equivalent to 4.98—10.13 ng/mL based on TBF content) was significantly superior to that of HQD-SAN (MIC value 1.56—3.13 mg/mL) and TBF alone (MIC value 0.06—0.50 µg/mL) (P < 0.05). Conclusion The BBD successfully optimized the formulation and preparation process of TBF-HQD-SAN NPs. The resulting NPs demonstrated uniform particle size, excellent drug-loading performance, and significantly enhanced antifungal efficacy, laying a foundation for further research.

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江西省卫生健康委科技计划（202510005）;江西省自然科学基金青年项目（20232BAB216124）;赣鄱俊才支持计划-高层次高技能领军人才培养工程（赣人社字[2024]69号）