Effect of the synthetic coumarin, ethyl 2-oxo-2H-chromene-3-carboxylate, on activity of Crotalus durissus ruruima sPLA2 as well as on edema and platelet aggregation induced by this factor

Fonseca F.V.; Baldissera L.; Camargo E.A.; Antunes E.; Diz-Filho E.B.S.; Correa A.G.; Beriam L.O.S.; Toyama D.O.; Cotrim C.A.; Toyama M.H.

Effect of the synthetic coumarin, ethyl 2-oxo-2H-chromene-3-carboxylate, on activity of Crotalus durissus ruruima sPLA2 as well as on edema and platelet aggregation induced by this factor

Tipo

Artigo

Data de publicação

2010

Periódico

Toxicon

Citações (Scopus)

12

Autores

Fonseca F.V.
Baldissera L.
Camargo E.A.
Antunes E.
Diz-Filho E.B.S.
Correa A.G.
Beriam L.O.S.
Toyama D.O.
Cotrim C.A.
Toyama M.H.

Resumo

We show that ethyl 2-oxo-2H-chromene-3-carboxylate (EOCC), a synthetic coumarin, irreversibly inhibits phospholipase A2 (sPLA2) from Crotalus durissus ruruima venom (sPLA2r) with an IC50 of 3.1 ± 0.06 nmol. EOCC strongly decreased the Vmax and Km, and it virtually abolished the enzyme activity of sPLA2r as well as sPLA2s from other sources. The edema induced by sPLA2r + EOCC was less than that induced by sPLA2r treated with p-bromophenacyl bromide, which was more efficient at neutralizing the platelet aggregation activity of native sPLA2r. Native sPLA2r induced platelet aggregation of 91.54 ± 9.3%, and sPLA2r + EOCC induced a platelet aggregation of 18.56 ± 6.5%. EOCC treatment also decreased the myotoxic effect of sPLA2r. Mass spectrometry showed that EOCC formed a stable complex with sPLA2r, which increased the mass of native sPLA2r from 14,299.34 Da to 14,736.22 Da. Moreover, the formation of this complex appeared to be involved in the loss of sPLA2r activity. Our results strongly suggest that EOCC can be used as a pharmacological agent against the sPLA2 in Crotalus durissus sp. venom as well as other sPLA2s. © 2010 Elsevier Ltd.