Desenvolvimento, caracterização e avaliação de nanopartículas contendo curcumina e metotrexato em células de adenocarcinoma de pulmão (CALU-3)

Abstract

Curcumin (CUR) is a polyphenolic compound extracted from the rhizomes of Curcuma longa L. and has several biological effects, such as anti-inflammatory, antioxidant and antitumor. Methotrexate (MTX) is an antimetabolite drug, very indicated for the treatment of cancer and has the disadvantage of suffering efflux by the P-glycoprotein (gp P). In order to improve undesirable characteristics such as low aqueous solubility and reduced bioavailability, CUR and MTX are models of substances to be nanoencapsulated. The objective of this work was to develop and characterize polymeric nanoparticles containing CUR and MTX and to evaluate the synergistic function of the two compounds in lung adenocarcinoma cells (CALU-3). Seven formulations (NCUR-1, NCUR-2, NMTX-1, NMTX-2, NCM-1, NCM-1 and NCM-2) were developed from poly (ε-caprolactone) (PCL) and polyethylene glycol 6000 (PEG 6000) by the interfacial deposition method of the preformed polymer. Calu-3 tumor cell lines and normal NHI-3T3 cells were used for assessing viability. Nanoparticles with sizes (278 to 325 nm), pH (5.26 to 6.32), zeta potentials (-33 to - 41 mV) and suitable polydispersion indexes (0.237 to 0.341) were obtained. An analytical method was developed and validated by high performance liquid chromatography (HPLC) for quantification of the drugs in the nanocarreadores. The method was specific, linear, accurate, accurate and robust, in the range of 10 to 70 μg.mL-1 for MTX and 10 to 90 μg.mL-1 for CUR. All formulations were spherical and nanoscale. No change was observed in the functional groups of the drugs in the infrared spectra (FTIR). Thermal analysis (DSC and TGA) and X-ray diffraction (XRD) indicated that the incorporation of MTX and CUR into nanoparticles led to the amorphization of the drugs. In vitro studies using the CALU-3 strain show that the NCM-2 formulation, using MTX (0.3 mg / mL) and CUR (0.5 mg / mL) was more interesting, inferring that the CUR performed the inhibitory role of P gp, allowing the entry of MTX leading to cell death by its cytotoxic effect. Fluorescence morphological analysis showed a predominance of early apoptosis and late apoptosis for the formulation containing NCM-2. It is concluded that the NCM-2 formulation presented synergistic effect by the association of MTX and CUR.