Preparation and in vitro evaluation of co-loaded nanostructured lipid carriers for methotrexate and cucurbitacin B

LI Hao; BAO Ting-qi; HUANG Xin-yi; ZHOU Jian-wen; MIAO Wen-kai; WU Li-hong; WANG Zi-xun; CAO Li-xin; HAN Cui-yan

doi:10.20251/j.cnki.1003-3734.2025.07.012

Abstract：Objective:In this study,a nanostructured lipid carrier(NLC)was loaded with methotrexate(MTX)and cucurbitacin B(CuB),resulting in the formulation MTX-CuB-NLC,which was optimized and evaluated in vitro.Methods:An in vitro high-performance liquid chromatography analysis method of MTX and CuB was established.The MTX-CuB-NLCs were prepared using a melt emulsification low-temperature curing method.A single-factor analysis was conducted by using encapsulation efficiency and drug loading as evaluation indicators,to identify the optimal prescription using Box-Behnken design response surface methodology.The particle size,polydispersity index(PDI),and Zeta potential of the formulation were evaluated by means of nanoparticle size analyzer.The appearance and morphology of the formulation were examined through transmission electron microscopy.And its storage stability was investigated by changes in particle size and Zeta potential.The in vitro drug release of the drug delivery system was assessed using the dialysis bag method.Results:The optimal formulation of MTX-CuB-NLC was as follows:MTX 0.7 mg,CuB 0.5 mg,mixed fatty acid glyceride 104 mg,ELP 96 mg,water phase volume 4 mL,and PVA 44 mg.The entrapment efficiencies were MTX(61.03±2.40)%,CuB(81.02±1.61)%.The drug loading capacities were MTX(0.25±0.02)%,CuB(0.23±0.02)%.The average particle size(44.13±1.40)nm,PDI(0.279±0.12),Zeta potential-(17.10±4.98)mV.Under transmission electron microscopy,the particles appeared quasi-circular with smooth surface and excellent dispersion.They showed favorable stability during storage over 14 days.The cumulative release of MTX and CuB in 48 h was(86.01±2.25)%and(73.72±1.97)%,respectively.Conclusion:In this study,MTX-CuB-NLC was successfully prepared.The preparation method was stable and sustained-release,which laid the foundation for the further study of the preparation.