RESUMEN
Thymidine phosphorylase (TP) is an enzyme that is up-regulated in a wide variety of solid tumors, including breast and colorectal cancers. It is involved in tumor growth and metastasis, for this reason it is one of the key enzyme to be inhibited, in an attempt to prevent tumor proliferation. However, it also plays an active role in cancer treatment, through its contribution in the conversion of the anti-cancer drug 5-fluorouracil (5-FU) to an irreversible inhibitor of thymidylate synthase (TS), responsible of the inhibition of the DNA synthesis. In this work, the intrinsic TP fluorescence has been investigated for the first time and exploited to study TP binding affinity for the unsubstituted 5-FU and for two 5-FU derivatives, designed to expose this molecule on liposomal membranes. These molecules were obtained by functionalizing the nitrogen atom with a chain consisting of six (1) or seven (2) units of glycol, linked to an alkyl moiety of 12 carbon atoms. Derivatives (1) and (2) exhibited an affinity for TP in the micromolar range, 10 times higher than the parent compound, irrespective of the length of the polyoxyethylenic spacer. This high affinity was maintained also when the compounds were anchored in liposomal membranes. Experimental results were supported by molecular dynamics simulations and docking calculations, supporting a feasible application of the designed supramolecular lipid structure in selective targeting of TP, to be potentially used as a drug delivery system or sensor device.
Asunto(s)
Antimetabolitos Antineoplásicos/metabolismo , Biología Computacional/métodos , Fluorescencia , Fluorouracilo/metabolismo , Liposomas/química , Fosfolípidos/metabolismo , Timidina Fosforilasa/metabolismo , Antimetabolitos Antineoplásicos/química , Sitios de Unión , Fluorouracilo/química , Humanos , Liposomas/metabolismo , Fosfolípidos/química , Timidina Fosforilasa/químicaRESUMEN
Previous investigations showed that the extent of DNA condensation and the efficiency in the transfection of liposomes formulated with 1,2-dimyristoyl-sn-glycero-phosphocholine and cationic stereomeric gemini surfactants depend heavily on the stereochemistry of the gemini. The influence of the stereochemistry on the interaction of lipoplexes with zwitterionic and anionic cell membrane models was investigated by differential scanning calorimetry to rationalize their different biological behavior. Further, the thermotropic behavior of the corresponding liposomes and of the spontaneous self-assemblies of gemini surfactants in the presence and in the absence of DNA was evaluated to correlate the physicochemical properties of lipoplexes and the stereochemistry of the cationic component. The obtained results show that the stereochemistry of the gemini surfactant controls lipoplexes organization and their mode and kinetic of interaction with different cell membrane models.
Asunto(s)
Calcitriol/análogos & derivados , Liposomas/química , Modelos Químicos , Tensoactivos/química , Calcitriol/química , Rastreo Diferencial de Calorimetría , Cationes/química , ADN/química , Dimiristoilfosfatidilcolina/química , Estereoisomerismo , Temperatura de TransiciónRESUMEN
Lipid-based drug carriers, such as liposomes or drug/lipid complexes, have been extensively investigated in a large number of therapeutic protocols such as gene therapy, drug delivery, drug targeting and antibacterial treatments, in preclinical and clinical trials. Many formulations composed of natural and/or synthetic amphiphiles have been studied. Many synthetic lipids and surfactants have been designed and tested in order to improve liposomes and lipid complexes performances, such as fusion with cellular membrane, cellular uptake, target selectivity, transfection efficiency, low toxicity. Among these, gemini surfactants have been shown to be highly effective in delivering genetic material to cells, and also have been shown promising as synthetic additives in liposome formulations for drug delivery. The encouraging results obtained in gene therapy have given impulse to chemist creativity: an extensive selection of pH sensitive, sugar-, aminoacid- , and peptide-based gemini surfactants have been developed, many of which have shown good biological features. This review focuses on recent progress in gemini surfactant based formulations and their applications in different therapeutic protocols.