A Novel Nanoemulsion Formula for an Improved Delivery of a Thalidomide Analogue to Triple-Negative Breast Cancer; Synthesis, Formulation, Characterization and Molecular Studies

Noran M Tawfik,1,2 Mohammed S Teiama,3,4 Sameh Samir Iskandar,5 Ahmed Osman,1,6 Sherif F Hammad7,8 1Biotechnology Program, Basic and Applied Sciences Institute, Egypt-Japan University of Science and Technology, Alexandria, Egypt; 2Department of Zoology, Faculty of Science, Suez Canal University, Ismailia, Egypt; 3Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Helwan University, Cairo, Egypt; 4Department of Pharmaceutics, Faculty of Pharmacy, Galala University, Suez, Egypt; 5Fellow and Head of Surgical Oncology Department, Ismailia Teaching Oncology Hospital (GOTHI), Ismailia, Egypt; 6Department of Biochemistry, Faculty of Science, Ain Shams University, Cairo, Egypt; 7PharmD Programs, Egypt-Japan University of Science and Technology, Alexandria, Egypt; 8Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Helwan University, Cairo, EgyptCorrespondence: Noran M Tawfik; Ahmed Osman, Biotechnology Program, Basic and Applied Sciences Institute, Egypt-Japan University of Science and Technology, New Borg El-Arab, Alexandria, 21934, Egypt, Email noran.tawfik@ejust.edu.eg; ahmed.osman@ejust.edu.egBackground: Thalidomide (THD) and its analogues were recently reported as a promising treatment for different types of solid tumors due to their antiangiogenic effect.Methods: In this work, we synthesized a novel THD analogue (TA), and its chemistry was confirmed with different techniques such as IR, mass spectroscopy, elemental analysis as well as 1H and 13C NMR. To increase solubility and anticancer efficacy, a new oil in water (O/W) nanoemulsion (NE) was used in the formulation of the analogue. The novel formula’s surface charge, size, stability, FTIR, FE-TEM, in vitro drug release and physical characteristics were investigated. Furthermore, molecular docking studies were conducted to predict the possible binding modes and molecular interactions behind the inhibitory activities of the THD and TA.Results: TA showed a significant cytotoxic activity with IC50 ranging from 0.326 to 43.26 μmol/mL when evaluated against cancerous cells such as MCF-7, HepG2, Caco-2, LNCaP and RKO cell lines. The loaded analogue showed more potential cytotoxicity against MDA-MB-231 and MCF-7-ADR cell lines with IC50 values of 0.0293 and 0.0208 nmol/mL, respectively. Moreover, flow cytometry of cell cycle analysis and apoptosis were performed showing a suppression in the expression levels of TGF-β, MCL-1, VEGF, TNF-α, STAT3 and IL-6 in the MDA-MB-231 cell line.Conclusion: The novel NE formula dramatically reduced the anticancer dosage of TA from micromolar efficiency to nanomolar efficiency. This indicates that the synthesized analogue exhibited high potency in the NE formulation and proved its efficacy against triple-negative breast cancer cell line.Graphical Abstract: Keywords: thalidomide analogue, antiangiogenic, nanoemulsion, cytotoxicity, triple-negative breast cancer, docking study