RESUMO
Methotrexate (MTX) is the most important drug used in the treatment of several kinds of cancers, such as colon cancer. However, this drug can cause a reduction in the target tissue bioavailability. It is administered orally and absorbed quickly. This study aimed to produce an anti-colon cancer prodrug based on MTX via loading it into a biopolymer compound. Chitosan (CS) was extracted from scales of local fish by utilizing a previously published protocol. The MTX was then transformed to Methotrexate - imidazole and loaded into CS to prepare Chitosan - Methotrexate (CS-MTX) conjugates as colon cancer prodrugs. Fourier-transform infrared (FTIR), UV-visible spectroscopy, and 1H-NMR were used to analyse the structure of the prepared compounds. The prepared compounds were also tested for hemolytic activity. Chemical stability was studied using 0.2 M from the different buffer types with a pH of 1.2 and 7.4 over different periods about 240 min and kept in an incubator at 37 °C. The loading percentage was measured by hydrolysing the amide bond in basic media followed by the measurement of the absorbency at 273 nm. Three types of cancer cells, MCF-7, MDA-MB-231, and MDA-MB-453, were used to test the anticancer effects of CS-MTX by using tetrazolium bromide (MTT) assay. The results indicated that the viability of human breast cancer cell lines decreased because of the use of CS-MTX. This study also showed that CS-MTX was less toxic than the original drug. Therefore, it may be measured for additional biological analyses and medical applications. The results presented here showed that the new compound is remarkably stable in comparison with MTX and has longer half-life (t ½). Therefore, the CS-MTX has promising strategies through minimising the side effects of anti-colon tumour drugs.
RESUMO
The cell wall of mycobacteria is characterised by glycolipids composed of different classes of mycolic acids (MAs; alpha-, keto-, and methoxy-) and sugars (trehalose, glucose, and arabinose). Studies using mutant Mtb strains have shown that the structure of MAs influences the inflammatory potential of these glycolipids. As mutant Mtb strains possess a complex mixture of glycolipids, we analysed the inflammatory potential of single classes of mycolate esters of the Mtb cell wall using 38 different synthetic analogues. Our results show that synthetic trehalose dimycolate (TDM) and trehalose, glucose, and arabinose monomycolates (TMM, GMM, and AraMM) activate bone marrow-derived dendritic cells in terms of the production of pro-inflammatory cytokines (IL-6 and TNF-α) and reactive oxygen species, upregulation of costimulatory molecules, and activation of NLRP3 inflammasome by a mechanism dependent on Mincle. These findings demonstrate that Mincle receptor can also recognise pentose esters and seem to contradict the hypothesis that production of GMM is an escape mechanism used by pathogenic mycobacteria to avoid recognition by the innate immune system. Finally, our experiments indicate that TMM and GMM, as well as TDM, can promote Th1 and Th17 responses in mice in an OVA immunisation model, and that further analysis of their potential as novel adjuvants for subunit vaccines is warranted.
