In vitro and In vivo evaluation of 2-amino-7(8)-fluorophenazine N5,N10-dioxide within polymeric micelles as theranostic agents

N. Lecot
Centro de Investigaciones Nucleares. Facultad de Ciencias, Universidad de la República, Montevideo, Uruguay.,
Uruguay

Keywords: breast cancer, hypoxic, fluorophenazine, polymeric micelles, Ames test, theranostic agents

Summary:

In vitro and In vivo evaluation of 2-amino-7(8)-fluorophenazine N5,N10-dioxide within polymeric micelles as theranostic agents Nicole Lecot1,2, Belén Dávila1, Carina Sánchez1, Marcelo Fernandez3, Joaquin Afonso2, Mirel Cabrera2, Hugo Cerecetto1,2 Pablo Cabral2, Romina Glisoni4, Mercedes González1 1. Grupo de Química Orgánica Medicinal, Facultad de Ciencias, Universidad de la República, Iguá 4225, Montevideo, 11400, Uruguay. 2. Área de Radiofarmacia, Centro de Investigaciones Nucleares, Facultad de Ciencias, Universidad de la República, Iguá 4225, Montevideo, 11400, Uruguay. 3. Laboratorio de Experimentación Animal, Centro de Investigaciones Nucleares, Facultad de Ciencias, Universidad de la República, Iguá 4225, Montevideo, 11400, Uruguay. 4. Instituto NANOBIOTEC UBA-CONICET. Departamento de Tecnología Farmacéutica, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina. Contact e-mail: lecotn@gmail.com Breast cancer, one of the most common cancer in developing countries, is reported to feature high incidence, relatively high morbidity and mortality in women since the diagnosis and treatment is made in the later stages of the disease. It is a solid tumor characterized by a high level of hypoxic areas which are difficult to treat. We have developed bioreducible pro-drugs under hypoxic conditions in which 2-amino-7(8)-fluorophenazine N5,N10-dioxide (FNZ) stands out as a potent and selective anticancer agent (1). To improve its solubility and stability, the encapsulation was studied using pristine polymeric micelles (PMs) and glycosylated derivatives (2) as a strategy to: (a) increase the solubility, (b) stabilize the aqueous formulation for intravenous application and (c) have nanoscale size which is suitable for the enhanced permeability and retention (EPR) effect for cancer diagnosis and improved antitumor activity (3). This work presents the characterization, encapsulation and the In vitro-In vivo evaluation. We have studied the physicochemical characterization of the nanosystems and the evaluation of the mutagenic properties in vitro (AMES test) (4) and its potential activity as an anti-cancer agent of FNZ and PMs/FNZ(5). Finally we explored its property as a diagnostic agent (99Tc-PMs/FNZ). The preliminary results indicate that all the systems under study (Free-FNZ, pristine PMs/FNZ and glycosylated PMs/FNZ) were stable in particle size and Z-potential after its resuspension in ultrapure water. The FNZ and PMs/FNZ exhibited mutagenicity in the TA 98 strain, while in the TA 100 PM-Glu/FNZ (F127-Glu/FNZ) did not. The In vivo studies reveal an enhanced circulation time and a good performance of a theranostic agent in BALB/c mice with an induced 4T1 murine mammary tumor cell line. References: (1) Cerecetto H, et al. Phenazine 5, 10-dioxide derivates as hypoxic selective cytotoxins:Part II Structure-activity relationships studies. MedChem. 2006; 5, 511-521. (2) Glisoni RJ, et al. Novel Poly(Ethylene Oxide)-b-Poly(Propylene Oxide) Copolymer-Glucose Conjugate by the Microwave-Assisted Ring Opening of a Sugar Lactone. Macromolecular Bioscience. 2014; 14, 1639-1651. (3) Torchilin V, et al. Tumor delivery of macromolecular drugs based on the EPR effect. Advanced Drug Delivery Reviews. 2011; 63,131-135.(4) OECD Guide Bacterial Reverse Mutation Test. (5) Oerlemans C, , et al. Polymeric Micelles in Anticancer Therapy: Targeting, Imaging and Triggered Release. Pharm Res. 2010; 27(12): 2569–2589. Acknowledgement: The authors want to thank the ANII for financial support