LADAF, Department of Food and Experimental Nutrition, Faculty of Pharmaceutical Sciences, University of São Paulo, São Paulo, Brazil.
Department of Organic Chemistry, Institute of Chemistry, Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil.
Eur J Pharm Biopharm. 2021 Feb;159:99-107. doi: 10.1016/j.ejpb.2020.12.016. Epub 2021 Jan 6.
Atherosclerosis is a non-resolving inflammatory condition that underlies major cardiovascular diseases.Recent clinical trial using an anti-inflammatory drug has showna reduction of cardiovascular mortality, but increased the susceptibility to infections. For this reason, tissue target anti-inflammatory therapies can represent a better option to regress atherosclerotic plaques. Docosahexaenoic acid (DHA) is a natural omega 3 fatty acidcomponentof algae oil and acts asaprecursor of several anti-inflammatory compounds, such the specialized proresolving lipid mediators(SPMs). During the atherosclerosis process, the inflammatory condition of the endothelium leads to the higher expression of adhesion molecules, such as Endothelial Cell Adhesion Molecule Plate 1 (PECAM-1 or CD31), as part of the innate immune response. Thus, the objective of this study was to develop lipid-core nanocapsules with DHA constituting the nucleus and anti-PECAM-1 on their surface and drive this structure to the inflamed endothelium. Nanocapsules were prepared by interfacial deposition of pre-formed polymer method. Zinc-II was added to bind anti-PECAM-1 to the nanocapsule surface by forming an organometallic complex. Swelling experiment showed that the algae oil act as non-solvent for the polymer (weight constant weight for 60 days, p > 0.428) indicating an adequate material to produce kinetically stable lipid-core nanocapsules (LNC). Five formulations were synthesized: Lipid-core nanocapsules containing DHA (LNC-DHA) or containing Medium-chain triglycerides (LNC-MCT), multi-wall nanocapsules containing DHA (MLNC-DHA) or containing MCT (MLNC-MCT) and the surface-functionalized (anti-PECAM-1) metal-complex multi-wall nanocapsules containing DHA (MCMN-DHA-a1). All formulations showed homogeneous macroscopic aspects without aggregation. The mean size of the nanocapsules measured by laser diffraction did not show difference among the samples (p = 0.241). Multi-wall nanocapsules (MLNC) showed a slight increase in the mean diameter and polydispersity index (PDI) measured by DLS, lower pH and an inversion in the zeta-potential (ξP) compared to LNCs. Conjugation test for anti-PECAM-1 showed 94.80% of efficiency. The mean diameter of the formulation had slightly increased from 160 nm (LCN-DHA) and 162 nm (MLNC-DHA) to 164 nm (MCMN-DHA-a1) indicating that the surface functionalization did not induce aggregation of the nanocapsules. Biological assays showed that the MCMN-DHA-a1 were uptaken by the HUVEC cells and did not decrease their viability. The surface-functionalized (anti- PECAM-1) metal-complex multi-wall nanocapsules containing DHA (MCMN-DHA-a1) can be considered adequate for pharmaceutical approaches.
动脉粥样硬化是一种未解决的炎症性疾病，是主要心血管疾病的基础。最近使用抗炎药物的临床试验表明，心血管死亡率降低，但感染易感性增加。因此，组织靶向抗炎治疗可能是使动脉粥样硬化斑块消退的更好选择。二十二碳六烯酸（DHA）是一种天然的ω-3 脂肪酸，是藻类油的成分之一，作为几种抗炎化合物的前体，如特殊的促解决脂质介质（SPM）。在动脉粥样硬化过程中，内皮的炎症状态导致细胞间黏附分子的高表达，如内皮细胞黏附分子 1（PECAM-1 或 CD31），作为先天免疫反应的一部分。因此，本研究的目的是开发含有 DHA 的核芯脂质纳米囊，在其表面带有抗 PECAM-1，并将该结构驱动到炎症内皮。纳米囊通过界面沉积法制备。加入锌-II 通过形成有机金属络合物将抗 PECAM-1 结合到纳米囊表面。溶胀实验表明，藻类油作为聚合物的非溶剂（60 天内重量恒定，p > 0.428），表明有足够的材料来生产动力学稳定的核芯脂质纳米囊（LNC）。合成了五种配方：含 DHA 的核芯纳米囊（LNC-DHA）或含中链甘油三酯（LNC-MCT）、含 DHA 的多壁纳米囊（MLNC-DHA）或含 MCT（MLNC-MCT）和表面功能化（抗 PECAM-1）的金属络合物多壁纳米囊（MCMN-DHA-a1）。所有配方均表现出均匀的宏观外观，无聚集现象。激光衍射法测量的纳米囊的平均粒径在样品间无差异（p = 0.241）。多壁纳米囊（MLNC）的平均粒径和多分散指数（PDI）略有增加，通过 DLS 测量，pH 值降低，zeta 电位（ξP）反转与 LNCs 相比。抗 PECAM-1 的结合试验显示效率为 94.80%。配方的平均粒径从 160nm（LCN-DHA）和 162nm（MLNC-DHA）略有增加到 164nm（MCMN-DHA-a1），表明表面功能化没有引起纳米囊的聚集。生物测定表明，MCMN-DHA-a1 被 HUVEC 细胞摄取，并且不降低其活力。含 DHA 的表面功能化（抗 PECAM-1）金属络合物多壁纳米囊（MCMN-DHA-a1）可用于药物制剂。
