Svetlichny G, Külkamp-Guerreiro I C, Cunha S L, Silva F E K, Bueno K, Pohlmann A R, Fuentefria A M, Guterres S S
Pharmazie. 2015 Mar;70(3):155-64.
The aim of this work was to develop solid lipid nanoparticles (SLN) containing copaiba oil with and without allantoin (NCOA, NCO, respectively) and to evaluate their antifungal activity. Nanoparticle suspensions were prepared using a high homogenisation technique and characterised by dynamic light scattering, laser diffraction, nanoparticle tracking analysis, multiple light scattering analysis, high-pressure liquid chromatography, pH and rheology. The antifungal activities of the formulations were tested in vitro against the emergent yeasts Candida krusei and Candida parapsilosis, and the fungal pathogens of human skin Trichophyton rubrum and Microsporum canis. The dynamic light scattering analysis showed z-average diameters (intensity) between 118.63 ± 8.89 nm for the nanoparticles with both copaiba oil and allantoin and 126.06 ± 9.84nm for the nanoparticles with just copaiba oil. The D[4,3] determined by laser diffraction showed similar results of 123 ± 1.73 nm for the nanoparticles with copaiba oil and allantoin and 130 ± 3.6 nm for the nanoparticles with copaiba oil alone. Nanoparticle tracking analysis demonstrated that both suspensions had monomodal profiles and consequently, the nanoparticle populations were homogeneous. This analysis also corroborated the results of dynamic light scattering and laser diffraction, exhibiting a smaller mean diameter for the nanoparticles with copaiba oil and allantoin (143 nm) than for the nanoparticles with copaiba oil (204 nm). The physicochemical properties indicated that the dispersions were stable overtime. Rheology evidenced Newtonian behaviour for both suspensions. Antifungal susceptibility showed a MIC90 of 125 μg/mL (nanoparticles with copaiba oil) and 7.8 μg/mL (nanoparticles with copaiba oil and allantoin) against C. parapsilosis. The nanoparticles with copaiba oil and the nanoparticles with copaiba oil and allantoin presented a MIC90 of 500 μg/mL and 250 μg/mL, respectively, against C. krusei. The MIC90 values were 500 μg/mL (nanoparticles with copaiba oil) and 1.95 μg/mL (nanoparticles with copaiba oil and allantoin) against T. rubrum. Against M. canis, the nanoparticles with copaiba oil and allantoin had a MIC9 of 1.95 μg/mL. In conclusion, nanoencapsulation improved the antifungal activity of copaiba oil, which was enhanced by the presence of allantoin. The MICs obtained are comparable to those of commercial products and can represent promising therapeutics for cutaneous infections caused by yeasts and dermatophytes.
本研究旨在开发含有巴西香脂油（分别添加和不添加尿囊素，即NCOA和NCO）的固体脂质纳米粒（SLN），并评估其抗真菌活性。采用高均质技术制备纳米粒悬浮液，并通过动态光散射、激光衍射、纳米颗粒跟踪分析、多重光散射分析、高压液相色谱、pH和流变学对其进行表征。在体外测试了这些制剂对新兴酵母克柔念珠菌和近平滑念珠菌，以及人类皮肤真菌病原体红色毛癣菌和犬小孢子菌的抗真菌活性。动态光散射分析显示，同时含有巴西香脂油和尿囊素的纳米粒的z平均直径（强度）在118.63±8.89nm之间，仅含有巴西香脂油的纳米粒的z平均直径为126.06±9.84nm。激光衍射测定的D[4,3]显示，含有巴西香脂油和尿囊素的纳米粒的结果相似，为123±1.73nm，仅含有巴西香脂油的纳米粒为130±3.6nm。纳米颗粒跟踪分析表明，两种悬浮液均具有单峰分布，因此纳米颗粒群体是均匀的。该分析还证实了动态光散射和激光衍射的结果，显示含有巴西香脂油和尿囊素的纳米粒的平均直径（143nm）小于仅含有巴西香脂油的纳米粒（204nm）。物理化学性质表明，这些分散体随时间是稳定的。流变学证明两种悬浮液均表现出牛顿行为。抗真菌药敏试验显示，针对近平滑念珠菌，含有巴西香脂油的纳米粒的MIC90为125μg/mL，含有巴西香脂油和尿囊素的纳米粒为7.8μg/mL。针对克柔念珠菌，含有巴西香脂油的纳米粒和含有巴西香脂油和尿囊素的纳米粒的MIC90分别为500μg/mL和250μg/mL。针对红色毛癣菌，MIC90值分别为500μg/mL（含有巴西香脂油的纳米粒）和1.95μg/mL（含有巴西香脂油和尿囊素的纳米粒）。针对犬小孢子菌，含有巴西香脂油和尿囊素的纳米粒的MIC9为1.95μg/mL。总之，纳米包封提高了巴西香脂油的抗真菌活性，尿囊素的存在进一步增强了这种活性。所获得的MIC与商业产品相当，可代表治疗由酵母和皮肤癣菌引起的皮肤感染的有前景的疗法。
