Zaki Randa Mohammed, Aldosari Basmah Nasser, Alkharashi Layla A, Alsalhi Alyaa, Abo El-Ela Fatma I, Alosaimi Raneem Meshal, Alsunbul Maha, Afzal Obaid, Said Mayada
Department of Pharmaceutics, College of Pharmacy, Prince Sattam Bin Abdulaziz University, Al-Kharj, Saudi Arabia.
Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Beni-Suef University, Beni-Suef, Egypt.
PLoS One. 2025 Jul 17;20(7):e0325511. doi: 10.1371/journal.pone.0325511. eCollection 2025.
The goal of this study was the formulation and optimization by statistical means of bilosomal formulations of axitinib (AXT) in order to improve its anticancer efficacy in a targeted manner. A central composite rotatable design was employed Using Design-Expert® software. The formulation factors were cholesterol, span 60, and sodium deoxy cholate (SDC) amounts (mg), whereas the dependent responses were Entrapment efficiency (EE%), Vesicles' size (VS), and Zeta potential (ZP). The design expert software was utilized to perform the numerical optimization process. The optimized bilosomal formulation was assessed using differential scanning calorimetry (DSC), X-ray diffraction (XRD), transmission electron microscope (TEM), in-vitro release study, short-term stability study, and in-vitro cell proliferation assay and flow cytometry on MCF-7 breast and OV-2774 ovarian cancer cell lines. The optimized formulation was found to be composed of 19.999, 111.869 and 15 mgs of cholesterol, span 60, and SDC, respectively with a desirability of 0.753. EE%, VS, and ZP were predicted to be 88.4977%, 594.592 nm, and -44.2354 mV, respectively. The validation process on the optimized formula demonstrated that the variation from the predicted responses was less than 5%. The DSC and XRD studies revealed that AXT was entrapped within the bilosomal vesicles. The optimized AXT bilosomal formulation exhibited spherical non-aggregated nanovesicles in TEM images. Furthermore, it improved AXT release when compared to AXT suspension. According to stability experiments, the optimum bilosomal formulation was stable for thirty days. The cytotoxicity of the optimized bilosomal formulation was enhanced on the MCF-7 breast and OV-2774 ovarian cancer cell lines compared to AXT suspension even at lower concentrations. Flow cytometry showed that AXT loaded BSMs made a significant increase in the percentage of apoptotic cells in MCF-7 and OV-2774 cells, respectively. Molecular docking suggests that axitinib and SDC decreased the activation of the caspase-8 receptor on the surface of ovarian and breast cancer, which consequently led to an increase in anticancer activity. So, BSMs might be regarded a promising carrier of AXT to target ant treat breast and ovarian cancers.
本研究的目的是通过统计学方法制备和优化阿昔替尼（AXT）的双分子层脂质体剂型，以靶向方式提高其抗癌疗效。采用中心复合旋转设计，使用Design-Expert®软件。处方因素为胆固醇、司盘60和脱氧胆酸钠（SDC）的用量（mg），而相关响应指标为包封率（EE%）、囊泡大小（VS）和zeta电位（ZP）。利用该软件进行数值优化过程。使用差示扫描量热法（DSC）、X射线衍射（XRD）、透射电子显微镜（TEM）、体外释放研究、短期稳定性研究以及对MCF-7乳腺癌细胞系和OV-2774卵巢癌细胞系进行体外细胞增殖测定和流式细胞术，对优化后的双分子层脂质体剂型进行评估。发现优化后的剂型分别由19.999、111.869和15mg的胆固醇、司盘60和SDC组成，可取性为0.753。预测EE%、VS和ZP分别为88.4977%、594.592nm和-44.2354mV。对优化配方的验证过程表明，与预测响应的偏差小于5%。DSC和XRD研究表明AXT被包封在双分子层脂质体囊泡内。优化后的AXT双分子层脂质体剂型在TEM图像中呈现出球形、无聚集的纳米囊泡。此外，与AXT悬浮液相比，它改善了AXT的释放。根据稳定性实验，最佳双分子层脂质体剂型在30天内稳定。与AXT悬浮液相比，即使在较低浓度下，优化后的双分子层脂质体剂型对MCF-7乳腺癌细胞系和OV-2774卵巢癌细胞系的细胞毒性也增强。流式细胞术显示，负载AXT的双分子层脂质体分别使MCF-7和OV-2774细胞中的凋亡细胞百分比显著增加。分子对接表明，阿昔替尼和SDC降低了卵巢癌和乳腺癌表面半胱天冬酶-8受体的激活，从而导致抗癌活性增加。因此，双分子层脂质体可能被视为AXT靶向治疗乳腺癌和卵巢癌的有前景的载体。
