Combinatorial Anti-Mitotic Activity of Loratadine/5-Fluorouracil Loaded Zein Tannic Acid Nanoparticles in Breast Cancer Therapy: In silico, in vitro and Cell Studies

Mohamed Hamdi,1,2 Moawia M Al-Tabakha,3,4 Isra H Ali,1,5 Islam A Khalil6 1Department of Pharmaceutics, Faculty of Pharmacy, University of Sadat City, Sadat City, Egypt; 2Nanomedicine Laboratory, Faculty of Pharmacy, University of Sadat City, Sadat City, Egypt; 3Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, Ajman University, Ajman, United Arab Emirates; 4Centre of Medical and Bio-Allied Health Sciences Research Centre, Ajman University, Ajman, United Arab Emirates; 5Department of Cardiovascular Medicine, Mayo Clinic, Rochester, MN, USA; 6Department of Pharmaceutics, College of Pharmaceutical Sciences and Drug Manufacturing, Misr University for Science and Technology, 6th of October City, Giza, 12582, EgyptCorrespondence: Islam A Khalil, Department of Pharmaceutics, College of Pharmaceutical Sciences and Drug Manufacturing, Misr University of Science and Technology, 6th of October City, Giza, 12582, Egypt, Tel +201090140748, Email islam.khalil@must.edu.eg; islamkhll@gmail.com Moawia M Al-Tabakha, Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, Ajman University, P.O. Box 346, Ajman, United Arab Emirates, Tel +97167056208, Email m.altabakha@ajman.ac.aeIntroduction: Chemotherapy, the first approach in breast cancer management, is limited owing to systemic toxicity and drug resistance. For instance, 5-fluorouracil in recommended doses cause severe side effects, highlighting the urgent necessity of finding more effective and safer combinations. Hence, this study aims to develop biocompatible natural-based nanocarriers for the co-delivery of loratadine, an antihistaminic drug along with 5-fluorouracil in order to enhance the anticancer efficacy while reducing the required dose of 5-fluorouracil.Methods: In silico virtual screening was performed to examine the probable molecular interactions between loratadine or 5-fluorouracil, individually with two different polymers, chitosan and zein, to determine the most suitable carrier system. Zein exhibited superior binding affinity compared to chitosan. Nanoparticle optimization was conducted using a Box-Behnken design with zein, tannic acid, and either loratadine or 5-fluorouracil concentration as independent variables. The optimized formulations were characterized by dynamic light scattering, entrapment efficiency, morphology, in-vitro release, followed by cytotoxicity, apoptosis, and cell-cycle analyses in MCF-7 cells.Results: The optimal formulation consisted of zein (50 mg), tannic acid (131.93 mg), and loratadine or 5-fluorouracil (5 mg). The optimized formulation of Loratadine loaded nanoparticles (NPs) showed a particle size of 197 nm, polydispersity index (PDI) of 0.153, zeta potential of – 21.78 mV, and entrapment efficiency of 61.33%. Furthermore, the optimized 5-fluorouracil loaded nanoparticles exhibited a particle size of 231 nm, 0.170 for PDI, zeta potential of – 24.01 mV, and EE of 74.91% for entrapment efficiency. The sustained drug release profile exhibited a controlled pattern over 24– 48 h. Flow cytometry results showed that the mixed nanoparticles exhibited potent cytotoxicity equivalent to 5-fluorouracil loaded nanoparticles alone despite containing only half the 5-fluorouracil dose, confirming a potential synergistic effect.Conclusion: These findings confirmed the potential of drug-loaded nanoparticles as promising drug delivery systems for breast cancer management.Keywords: in silico, loratadine, 5-fluorouracil, zein, tannic acid