Título

PROSPECTION OF NEW ANTI-ANGIOGENIC DRUGS BASED ON CHEMICALLY MODIFIED HEPARINS

Introdução / Purpose

Angiogenesis is the formation of new blood vessels from preexisting vasculature. Pathologic angiogenesis in the eye such as choroidal neovascularization secondary to wet age-related macular degeneration lead to severe visual impairment and dramatically affect the quality of life. The search for emerging therapies to treat neovascularization pointed heparin as good target since it has important functions as regulator of vessel growth. The purpose of this study is investigate the anti-angiogenic effect of chemically modified heparins (mHep) in vitro.

Material e Método / Methods

Three mHEP were used in this study; O-N-desulfated (O-N); 2-0-desulfated (2-0); N-desulfated Re-N-acetylated (N-DRN) with no anti-coagulant or hemorrhagic effects. After 24 hours of treatment with mHep at 10ng/ml, 100ng/ml and 1000ng/ml or saline (PBS) the following tests were performed: Proliferation test in endothelial cells (EC) and the evaluation by direct cell counting, Anti-angiogenic assay by matrigel , cell viability by MTT test in EC and ARPE-19, EC migration assay in transwell. EC adhesion assay using fibronectin, laminin and gelatin as substrates.

Resultados / Results

All mHep showed anti-proliferative effets in ECs after 24 hours of treatment. Dose-related effect was observed with better results in 1000ng/ml. The anti-angiogenic test in matrigel showed an inhibition in tube formation after the treatment with the 2-0 and N-DRN. EC or ARPE-19 cell viability was not affected by the the treatment with mHEPs on the different concentrations. The migration of EC was reduced after the treatment with mHeps 2-0 and N-DRN. Adhesion of EC on fibronectin, laminin and gelatin was reduced after treatment with O-N and 2-0.

Discussão e Conclusões / Discussion

These three mHep have shown anti-angiogenic, anti-proliferative, anti-adhesive and anti-migratory effects on ECs but no effects on ARPE-19 retinal cells viability. In vivo studies using choroidal neovascular models will be used to further investigate the efficacy of these new molecules with anti-angiogenic potential.

Palavras Chave

Angiogenesis, new therapies, modified heparins