Caracterização estrutural e atividades farmacológicas do alginato obtido da alga Dictyopteris delicatula (J.V. Lamouroux., 1809) e seu derivado sulfatado

Marine algae are rich sources of various structural compounds which recently has been increasingly studied as a new source of bioactive substances. The alginate, as come as fucans, are considered the main acidic polysaccharides found in brown seaweed. This molecule consists a linear natural polysaccharide, non-sulfated, and presents monosaccharides: acid β-D-mannuronic (M) and α-L-guluronic acid (G); in a vast amount compositions and threads. Alginate has been widely applied in food and pharmaceutical industries because of its ability to retain water, forming films and gels as well as thickening, stabilizing and form emulsions. In this work we aimed to extract, structurally characterize, compare and analyze the possible pharmacological activities of native alginate molecule obtained from brown seaweed Dyctiopteris delicatula (DYN), and its chemically sulfated derivative (DYS). The alginate structure and composition molecule can be proven through chemical dosing, that showed low protein contamination and high sugar level, existence and separation of M and G blocks in the descending paper chromatography, infrared spectroscopy and nuclear magnetic resonance. Molecule sulfation was proven with sulphate dosage, resulting in 28.56% sulphate in molecule; electrophoresis, verify metachromasia with toluidine blue; and infrared spectroscopy, that showed a characteristic band at 1221cm-1 corresponding a sulfate group vibration. For the pharmacological activities the tests was: antioxidant activity, changes in cell function (MTT test) and anticoagulant test. In the antioxidant activity we observed that DYN showed better results in the kidnapping of hydroxyl radicals and ferric chelation compared to DYS, this had the best result in the total antioxidant capacity. Both showed similar activity in reducing power and the kidnapping radicals DPPH. In MTT test DYN and DYS had not proliferative and cytotoxic activity in fibroblast cells (3T3) and showed antiproliferative and cytotoxic activity in cancer cell lines HeLa and B16 melanoma. In anticoagulant assay DYN showed good activity in the intrinsic pathway of blood coagulation, and a small activity in the extrinsic pathway, in the other hand DYS showed only a very small activity in the extrinsic pathway, but cannot come to be regarded as an anticoagulant agent. From these results it can be concluded that the alginate was extracted and sulfated, revealing a potential compound to be used in the pharmaceutical industry as an anticoagulant agent, antioxidant and antitumor and the sulfation has not been conclusively important to performance in the tested pharmacological activities

Alginate is a linear polymer compound of D-mannuronic acid(M) linked (1 → 4) at its epimer in C-5 of L-guluronic acid (G). An alginate was isolated and partially purified from brown seaweed D. delicatula (DYN). DYN was chemically sufated (DYS). DYS showed 22% of sulfation and the rate of total sugars was around 66 and 59% for DYN and DYS respectively. DYN DYS and showed a ratio M G of 0.86 and 1.1, respectively. IR signals at 1221cm-1 confirmed the sulfation of DYS. DYN presented anticoagulant activity about intrinsic coagulation pathway, while DYS did not alter the clotting times for aPTT and PT. DYN and DYS also showed antioxidant properties especially ferric chelation where DYN reached about 97% and the OH sequestration where both averaged 85%. In 3T3 cells, DYN indicated proliferation in 24h and 48h of incubation and no cytotoxicity. For tumor lines HeLa and b16, and DYN and DYS decreased significantly the cell viability, especially after 48 hours (about 71% of inhibition). Given the results obtained in this study, it is clear that the sulfation is not decisive for the activity of D. delicatula alginate. In general, DYS and DYN shown as potential compounds to be used pharmacologically, and also in the food industry for revealing be great antioxidant.