依诺沙星温敏凝胶的制备及抗菌活性研究

张雪; 刘杰; 郭嘉欣; 于波

doi:10.13748/j.cnki.issn1007-7693.20243466

摘要:

目的制备依诺沙星固体脂质纳米粒温敏凝胶(enoxacin solid lipid nanoparticles gel，ENX-SLN-Gel)并考察其体外抗菌活性。

方法采用乳化蒸发-低温固化法制备ENX-SLN，以包封率、载药量为考察指标，通过单因素实验和Box-Behnken响应面法优化制备工艺，以粒径、Zeta电位、差示扫描量热法、傅里叶变换红外光谱及X射线衍射法对其进行表征；采用冷溶法制备温敏凝胶并进行质量评价；考察体外释放行为及透皮效果；通过CCK8法考察细胞毒性并进行皮肤刺激性实验观察其皮肤用药安全性；抑菌圈实验比较ENX和ENX-SLN-Gel的体外抗菌活性。

结果优化后的ENX-SLN的包封率为(89.86±0.60)%，载药量为(4.20±0.03)%，平均粒径为(262.4±0.72)nm，PDI为(0.167±0.64)，Zeta电位为(−14.3±5.25)mV，DSC、FT-IR、XRD进一步验证了ENX-SLN的形成；所得凝胶的胶凝温度为(32.4±0.46)℃，pH值为(6.30±0.13)，SEM、流变学证明了ENX-SLN-Gel的形成；体外释放拟合方程为Q=17.29t^1/2+11.37(R²=0.9668)，符合Higuchi模型，具有一定的缓释效果。体外透皮实验表明，该制剂的透皮效果优于原料药凝胶，可促进依诺沙星的经皮吸收。ENX-SLN-Gel在0~200 μg·mL⁻¹浓度范围内时，细胞的存活率＞95%；皮肤刺激性实验显示，完整皮肤和破损皮肤组均未见红斑、水肿等刺激性反应，说明其安全性良好；ENX和ENX-SLN-Gel的抑菌圈直径分别为(9.6±0.7)mm，(14.5±1.0)mm。

结论 ENX-SLN-Gel制备工艺方法简单，合理可行，具有良好的抗菌效果，有望成为外用制剂的一种新途径。

Abstract:

OBJECTIVE To prepare enoxacin solid lipid nanoparticles gel(ENX-SLN-Gel) and to investigate its antibacterial activity in vitro.

METHODS ENX-SLN were prepared by emulsion evaporation low-temperature solidification method, with encapsulation efficiency and drug loading as evaluation indicators. The preparation process was optimized through single factor experiments and Box Behnken response surface methodology. The particles were characterized by particle size, Zeta potential, differential scanning calorimetry, fourier transform infrared spectroscopy, and X-ray diffraction; temperature sensitive gel was prepared by cold solution method and its quality was evaluated; investigating in vitro drug release behavior and transdermal efficacy; using CCK8 method to investigate cell toxicity and conducting skin irritation experiments to observe the safety of skin medication; compared the in vitro antibacterial activity of ENX and ENX-SLN-Gel through inhibition zone experiments.

RESULTS The encapsulation efficiency of the optimized ENX-SLN was (89.86±0.60)%, the drug loading was (4.20±0.03)%, the average particle size was (262.4±0.72)nm, the PDI was (0.167±0.64), and the Zeta potential was (−14.3±5.25)mV. DSC, FT-IR, and XRD further confirmed the formation of ENX-SLN; the gelling temperature of the obtained gel was (32.4±0.46)℃, the pH value was (6.30±0.13), SEM and Rheology had demonstrated the formation of ENX-SLN-Gel; the fitting equation for in vitro release was Q=17.29t^1/2+11.37(R²=0.9668), which conformed to the Higuchi model and had a certain sustained release effect. The transdermal experiment in vitro showed that the transdermal effect of the preparation was better than that of the bulk drug gel, which could promote the transdermal absorption of enoxacin. When the concentration range of ENX-SLN-Gel was 0−200 μg·mL⁻¹, the cell survival rate was＞95%. The skin irritation test showed that there were no irritating reactions such as erythema or edema observed in both intact and damaged skin groups, indicating good safety; the diameters of the inhibition zones for ENX and ENX-SLN-Gel were (9.6±0.7)mm and (14.5±1.0)mm, respectively.

CONCLUSION The preparation process of ENX-SLN-Gel is simple, reasonable and feasible, with good antibacterial effects, and is expected to become a new approach for topical formulations.

依诺沙星温敏凝胶的制备及抗菌活性研究

Preparation and Antibacterial Activity of Enoxacin Thermosensitive Gel