RESUMO
In this present study, the anti-IIa activity and the antitumor properties of a hybrid heparin/heparan sulfate-like compound (sH/HS) from Litopenaeus vannamei shrimp heads are related. In addition to inhibiting 90.7% of thrombin activity at the lowest tested concentration (0.5⯵g/mL), sH/HS compound stimulated the synthesis of antithrombotic heparan sulfate by endothelial cells in a dose-dependent manner. In vitro experiments demonstrated that the molecule from shrimp displayed a potent anti-angiogenic effect, reducing over 80% of the tubular structures formation at 50 and 100⯵g/mL. In addition, sH/HS compound was able to inhibit the migration of B16F10 cells at all tested concentrations without affecting the cell viability. Although the studied compound had no effect on the proliferation of such cells during a period of 24â¯h, it had a significant long-term anti-proliferative effect, reducing about 80% of colony formation and anchorage-independent growth at 50 and 100⯵g/mL concentrations. When its effectiveness was tested in vivo, it was demonstrated that sH/HS promoted a reduction of more than 90% of tumor growth. In the context of thromboembolic disorders associated with cancer, such findings make the sH/HS compound an excellent target for studies on inhibiting of development and tumor progression, and the prevention of coagulopathies.
Assuntos
Heparina/química , Heparitina Sulfato/química , Heparitina Sulfato/farmacologia , Penaeidae/química , Protrombina/antagonistas & inibidores , Inibidores da Angiogênese/química , Inibidores da Angiogênese/farmacologia , Animais , Antineoplásicos/química , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Endoteliais/citologia , Células Endoteliais/efeitos dos fármacos , CoelhosRESUMO
The structural characterization and the anticoagulant potential of a novel heparin/heparan sulfate-like compound from the heads of Litopenaeus vannamei shrimp are described. While it is distinct from either heparin or heparan sulfate, enzymatic depolymerization and nuclear magnetic resonance spectroscopy analyses revealed that this molecule does share some structural features with heparin, such as the high degree of N- and 6-O-sulfation and minor N-acetylation, and with heparan sulfate, in the glucuronic acid content. Its ability to stabilize human antithrombin explains its significant anticoagulant activity in aPTT and Factor-Xa inhibition assays. Interestingly, in contrast to mammalian heparin, the shrimp compound displayed negligible hemorrhagic effect. Together, these findings have particular interest since they reveal a novel molecule with significant anti-Xa activity coupled with low bleeding effects which make the shrimp heparin/HS-like compound a potential alternative for mammalian heparin.
Assuntos
Anticoagulantes/química , Hemorragia/prevenção & controle , Heparina/química , Heparitina Sulfato/química , Penaeidae/química , Acetilação , Animais , Anticoagulantes/isolamento & purificação , Anticoagulantes/farmacologia , Antitrombinas/antagonistas & inibidores , Antitrombinas/química , Antitrombinas/isolamento & purificação , Bovinos , Cromatografia por Troca Iônica , Fator Xa/química , Inibidores do Fator Xa , Ácido Glucurônico/química , Cabeça , Heparina/isolamento & purificação , Heparina/farmacologia , Heparitina Sulfato/isolamento & purificação , Heparitina Sulfato/farmacologia , Humanos , Intestinos/química , Pâncreas/química , Tempo de Tromboplastina Parcial , Ratos , Suínos , Cauda/irrigação sanguínea , Cauda/efeitos dos fármacosRESUMO
A natural heparin-like compound isolated from the crab Goniopsis cruentata was structurally characterized and its anticoagulant and hemorrhagic activities were determined. Enzymatic and nuclear magnetic resonance analysis revealed that its structure is rich in disulfated disaccharides, possessing significant amounts of 2-O-sulfated-ß-D-glucuronic acid units. Furthermore, low amounts of trisulfated disaccharide units containing 2-O-sulfated-α-L-iduronic acid were detected, when compared to mammalian heparin. In addition, this heparin-like structure showed negligible in vitro anticoagulant activity and low bleeding potency, facts that make it a suitable candidate for the development of structure-driven, heparin based therapeutic agents with fewer undesirable effects.
Assuntos
Anticoagulantes/farmacologia , Braquiúros/química , Glucuronatos/química , Heparina/farmacologia , Animais , Anticoagulantes/química , Anticoagulantes/isolamento & purificação , Configuração de Carboidratos , Sequência de Carboidratos , Bovinos , Dissacarídeos/química , Glucosamina/química , Glucuronatos/isolamento & purificação , Heparina/química , Heparina/isolamento & purificação , Heparitina Sulfato/química , Espectroscopia de Ressonância Magnética , Masculino , Peso Molecular , Tempo de Tromboplastina Parcial , Ratos , Ácidos Urônicos/químicaRESUMO
The anti-inflammatory properties of a heparin-like compound from the shrimp Litopenaeus vannamei are related. Besides reducing significantly (p<0.001) the influx of inflammatory cells to injury site in a model of acute inflammation, shrimp heparin-like compound was able to reduce the matrix metalloproteinase (MMPs) activity in the peritoneal lavage of inflamed animals. Moreover, this compound also reduced almost 90% the activity of MMP-9 secreted by human activated leukocytes. Negligible anti-coagulant activities in aPPT assay and a poor bleeding potential make this compound a better alternative than mammalian heparin as a possible anti-inflammatory drug.
Assuntos
Anti-Inflamatórios/farmacologia , Anticoagulantes/farmacologia , Glicosaminoglicanos/farmacologia , Heparina/farmacologia , Inflamação/tratamento farmacológico , Penaeidae/fisiologia , Animais , Anti-Inflamatórios/química , Anticoagulantes/química , Endotélio Vascular/citologia , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/metabolismo , Glicosaminoglicanos/química , Glicosaminoglicanos/isolamento & purificação , Hemorragia/tratamento farmacológico , Heparina/química , Heparina/isolamento & purificação , Heparitina Sulfato/metabolismo , Leucócitos/efeitos dos fármacos , Leucócitos/enzimologia , Metaloproteinase 9 da Matriz/metabolismo , Neutrófilos/efeitos dos fármacos , Cavidade Peritoneal/fisiologia , Coelhos , Ratos , SuínosRESUMO
Heparin, other sulphated glycosaminoglycans and histamine were extracted from various dissected organs of Anomalocardia brasiliana, a mollusc from the South Atlantic, and quantified. A good correlation between heparin and histamine content was found in the labial palp, intestine, ctenidium, mantle and foot tissues. The tissue location of metachromatic cells, putatively containing heparin, was identified histologically with Alcian Blue, Toluidine Blue, Masson trichrome, Haematoxylin-Eosin and PAS. Except for the foot, cells containing metachromatic granules were found in the epithelium surfaces of all the organs analysed. An in situ identification of heparin using nitrous acid and heparinase degradation has established unequivocally the presence of this compound in the metachromatic cells. The location of 'mast-like' cells at the epithelium surface of mollusc tissues exposed to the environment are very similar to the distribution of mammalian and other vertebrate mast cells and gives support to the suggestion for a role of mast cells in defense mechanisms.