Synthesis and characterization of phosphoethanolamine and its analogues: Comparison between synthetic phosphoethanolamine and liposomal analogues applied in breast cancer

Abstract

The Man's ability to replicate molecules of living beings, and to create other similar molecules, is a remarkable development in human history. Its birth dates back to 1828, when the German chemist Friedrich Wöhler synthesized urea molecule. Such molecules are usually known as natural products, a term which generally refers to secondary metabolites. Originally In the cancer treatment, analogous substances of natural molecules were employed. Regardless of the type of molecule, the treatment aims the interaction of the drug with the DNA, causing irreparable damages to it, thus avoiding the replication of new genetic material. Currently present drugs no longer interact with the DNA, but rather with the signalling of cells and / or the blocking of metabolic pathways of cancer cells. Thus, this work aims to synthesize and characterize molecules analogous of the synthetic phosphoethanolamine (Pho-S) and compare its efficacy with Pho-S studies and formulations associated with DODAC liposomes in the pro-apoptotic pathway of MCF-7 breast cancer cells. As well as using murine 4T1 tumor cells, to evaluate the efficacy of orthotopic antitumor response in vivo in Balb/C mice carrying breast cancer cells and treated with Pho-S, DODAC liposomal formulations and analogous compounds which exhibit anti-tumor characteristics in vitro. The synthetic changes of Pho-S will be made in the laboratory, changing the structural conformation, the increase of the lipid chain, the duplication (bis (2-aminoethyl) hydrogen phosphate) and triplication (tris (2-ammoniethyl) phosphate) of ethanolamine linked to the phosphate group and ligands addition the the Pho-S. Two sets of methodologies will be used, in vitro and in vivo. In the in vitro methodologies will be used the Pho-S, DODAC liposomes and Pho-S analogues in the human breast adenocarcinoma MCF-7 and 4T1 murine lines, the measurements of optical absorption, cytotoxicity evaluation, evaluation of apoptotic activity, necrosis, analysis of the electrical potential of the mitochondrial membrane by flow cytometry, the activities of BCL-X1, BCL-2, BAX, cytochrome c and caspase 3 and 9 by flow cytometry will also be evaluated. The in vivo methodologies will use female Balb/C mice, in which the tumour growth will be implanted and evaluated, these will be divided in 8 groups for the use of the different Pho-S formulations, DODAC liposome and analogues that presented positive responses in the in vitro methodology, such as evaluation of disease progression, side effects and pharmacokinetic aspects.