Alyaa Alsalhi

Fabrication and appraisal of targeted axitinib loaded bilosomes for the enhanced breast and ovarian anticancer activity

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.

Fabrication and appraisal of axitinib loaded PEGylated spanlastics against MCF- 7 and OV- 2774 cell lines using molecular docking methods and in-vitro study

Axitinib is a second-generation tyrosine kinase inhibitor that works by selectively inhibiting vascular endothelial growth factor receptors (VEGFR-1, VEGFR-2, VEGFR-3). Through this mechanism of action, axitinib blocks angiogenesis, tumor growth and metastases and therefor it shows significant promise as a chemotherapeutic agent for various types of cancer. Nevertheless, the clinical efficacy of this substance is hindered by its restricted solubility in water and inadequate stability. To address these challenges, we developed spanlastics with polyethylene glycol (PEG) to improve the efficacy and stability of axitinib against breast and ovarian tumor malignancies in a targeted manner. Moreover, the study conducts a thorough examination of the interactions between the ligand Axitinib alone or after coating with PEG and a diverse array of protein types in breast (Dopamine, VEGFR) and ovarian cancer (EGFR, BCL-xL). The fabrication of axitinib- spanlastics was achieved through a thin-film hydration method. The evaluation of the impact of formulation factors on the features of nanovesicles was conducted using the I- optimal design. Subsequently, the optimum formulation was calculated. The optimal formulation was coated with polyethylene glycol (axitinib-PEG-spanlastics). An in vitro assessment was computed to evaluate the efficiency of the optimized axitinib-PEG-spanlastics against the MCF-7 breast cancer cell line and the OV-2774 ovarian cancer cell line. The optimized axitinib-PEG-spanlastics formulation exhibited a diameter of 563.42 ± 8.63 nm, accompanied by a zeta potential of −46.44 ± 0.09 mV. The formulation demonstrated an 84.32 ± 3.64% entrapment percent and a cumulative release of 73.58 ± 3.37% during a 4-hour period. The results obtained from the WST-1 assay showed a significant decrease in the percentage of cell survival, reaching 50% at a concentration of 0.68 µM for the PEG-spanlastics. In contrast, the axitinib free drug suspension exhibited 50% cell survival at a concentration of 1.1 µM in the breast cancer (MCF-7) cell line. In MCF-7 cells, the percentage of apoptotic cells generated by axitinib-PEG-spanlastics compared to the free drug suspension was 70.76 ± 4.971% vs. 32.6 ± 1.803%, while in OV-2774 cells, it was 43.55 ± 4.243% vs. 24.44 ± 4.950%. These results propose that Axitinib-PEG-spanlastics have the potential to be a successful nanoplatform for targeting breast and ovarian cancer and effectively managing tumors.