Moreira Thais da Silva, Silva Alan Denis Olivindo, Vasconcelos Bianca Rodrigues Farias, Santos Elias da Silva, de Sousa Ana Carolina Cruz, de Freitas João Vito Barroso, de Oliveira Yara Santiago, Vidal Laura Maria Teodorio, Ribeiro Fábio de Oliveira Silva, de Araújo Alyne Rodrigues, Vieira Neto José de Brito, Pessoa Cláudia do Ó, Petrilli Raquel, Eloy Josimar O
Department of Pharmacy, Faculty of Pharmacy, Dentistry and Nursing, Federal University of Ceará, Fortaleza 60430-355, CE, Brazil.
Institute of Health Sciences, University of International Integration of the Afro-Brazilian Lusophony- UNILAB, Redenção 62790-970, CE, Brazil.
Pharmaceutics. 2023 Mar 11;15(3):915. doi: 10.3390/pharmaceutics15030915.
Docetaxel (DTX) is a non-selective antineoplastic agent with low solubility and a series of side effects. The technology of pH-sensitive and anti-epidermal growth factor receptor (anti-EGFR) immunoliposomes aims to increase the selective delivery of the drug in the acidic tumor environment to cells with EFGR overexpression. Thus, the study aimed to develop pH-sensitive liposomes based on DOPE (dioleoylphosphatidylethanolamine) and CHEMS (cholesteryl hemisuccinate), using a Box-Behnken factorial design. Furthermore, we aimed to conjugate the monoclonal antibody cetuximab onto liposomal surface, as well as to thoroughly characterize the nanosystems and evaluate them on prostate cancer cells. The liposomes prepared by hydration of the lipid film and optimized by the Box-Behnken factorial design showed a particle size of 107.2 ± 2.9 nm, a PDI of 0.213 ± 0.005, zeta potential of -21.9 ± 1.8 mV and an encapsulation efficiency of 88.65 ± 20.3%. Together, FTIR, DSC and DRX characterization demonstrated that the drug was properly encapsulated, with reduced drug crystallinity. Drug release was higher in acidic pH. The liposome conjugation with the anti-EGFR antibody cetuximab preserved the physicochemical characteristics and was successful. The liposome containing DTX reached an IC at a concentration of 65.74 nM in the PC3 cell line and 28.28 nM in the DU145 cell line. Immunoliposome, in turn, for PC3 cells reached an IC of 152.1 nM, and for the DU145 cell line, 12.60 nM, a considerable enhancement of cytotoxicity for the EGFR-positive cell line. Finally, the immunoliposome internalization was faster and greater than that of liposome in the DU145 cell line, with a higher EGFR overexpression. Thus, based on these results, it was possible to obtain a formulation with adequate characteristics of nanometric size, a high encapsulation of DTX and liposomes and particularly immunoliposomes containing DTX, which caused, as expected, a reduction in the viability of prostate cells, with high cellular internalization in EGFR overexpressing cells.
多西他赛（DTX）是一种具有低溶解度和一系列副作用的非选择性抗肿瘤药物。pH敏感型和抗表皮生长因子受体（抗EGFR）免疫脂质体技术旨在增加药物在酸性肿瘤环境中向EGFR过表达细胞的选择性递送。因此，本研究旨在采用Box-Behnken析因设计，开发基于二油酰磷脂酰乙醇胺（DOPE）和胆固醇半琥珀酸酯（CHEMS）的pH敏感型脂质体。此外，我们旨在将单克隆抗体西妥昔单抗偶联到脂质体表面，并全面表征纳米系统并在前列腺癌细胞上对其进行评估。通过脂质膜水化制备并经Box-Behnken析因设计优化的脂质体粒径为107.2±2.9nm，多分散指数（PDI）为0.213±0.005，ζ电位为-21.9±1.8mV，包封率为88.65±2.03%。综合傅里叶变换红外光谱（FTIR）、差示扫描量热法（DSC）和X射线衍射（DRX）表征表明，药物被正确包封，药物结晶度降低。在酸性pH条件下药物释放更高。脂质体与抗EGFR抗体西妥昔单抗的偶联保留了其物理化学特性且偶联成功。含DTX的脂质体在PC3细胞系中浓度为65.74nM时达到半数抑制浓度（IC），在DU145细胞系中为28.28nM。反过来，免疫脂质体对PC3细胞的IC为152.1nM，对DU145细胞系为12.60nM，对EGFR阳性细胞系的细胞毒性有显著增强。最后，在DU145细胞系中，免疫脂质体的内化速度比脂质体更快且程度更大，该细胞系中EGFR过表达更高。因此，基于这些结果，有可能获得一种具有适当纳米尺寸特性、DTX高包封率的制剂，以及含有DTX的脂质体，尤其是免疫脂质体，正如预期的那样，它们导致前列腺细胞活力降低，在EGFR过表达细胞中具有高细胞内化率。
