Cubosomes post-loaded with antimicrobial peptides: characterization, bactericidal effect and proteolytic stability
Cubosomes post-loaded with antimicrobial peptides: characterization, bactericidal effect and proteolytic stability
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DOI：
10.1016/j.ijpharm.2017.04.082
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发表时间：
2017-06-30
影响因子：
5.8
通讯作者：
Andersson, Martin
中科院分区：
文献类型：
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作者：
Boge, Lukas;Umerska, Anita;Andersson, Martin
近年来，抗菌肽(AMPS)等新型抗生素受到越来越多的关注。在本工作中，分散的立方体液晶凝胶(立方体)被用作三种AMPS(AP114、DPK-060和LL-37)的药物载体。用高效液相色谱、Zeta电位和圆二色谱研究了两种立方体肽比例下的多肽与立方体间的相互作用。用小角X射线散射和低温电子显微镜研究了AMPS对立方体结构的影响。采用最小抑菌浓度法和时间杀伤法研究了载AMP立方体体外抑菌作用。用两种弹性酶孵育的方法研究了其对蛋白质的保护作用，并用径向扩散法研究了蛋白质降解后的抑菌效果。观察到不同的AMP对立方体的结合效果不同，其中LL-37结合作用最强(>；60%)。负载AP114的立方体显示出保留的抗菌作用，而对于LL-37，广谱杀菌作用被减少到只包含革兰氏阴性菌。有趣的是，负载DPK-060的立方体对金黄色葡萄球菌和大肠杆菌有轻微的增强作用。此外，与未配制的多肽相比，这些立方体能保护LL-37免受蛋白质降解的影响，从而在受到弹性酶作用后具有显著更好的杀菌效果。(C)2017爱思唯尔B.V.保留所有权利。
Novel antibiotics, such as antimicrobial peptides (AMPS), have recently attended more and more attraction. In this work, dispersed cubic liquid crystalline gel (cubosomes) was used as drug delivery vehicles for three AMPs (AP114, DPK-060 and LL-37). Association of peptides onto cubosomes was studied at two cubosomeipeptide ratios using high performance liquid chromatography, zeta-potential and circular dichroism measurements. AMPs impact on the cubosome structure was investigated using small angle x-ray scattering and cryogenic transmission electron microscopy. The antimicrobial effect of the AMP loaded cubosomes was studied in vitro by minimum inhibitory concentration and time-kill assays. Proteolytic protection was investigated by incubating the,formulations with two elastases and the antimicrobial effect after proteolysis was studied using radial diffusion assay. Different association efficacy onto the cubosomes was observed among the AMPs, with LL-37 showing greatest association (>60%). AP114 loaded cubosomes displayed a preserved antimicrobial effect, whereas for LL-37 the broad spectrum bacterial killing was reduced to only comprise Gram-negative bacteria. Interestingly, DPK-060 loaded cubosomes showed a slight enhanced effect against S. aureus and E. coli strains. Moreover, the cubosomes were found to protect LL-37 from proteolytic degradation, resulting in a significantly better bactericidal effect after being subjected to elastase, compared to unformulated peptide. (C) 2017 Elsevier B.V. All rights reserved.