The Bioflavonoids Rutin and Rutin Succinate Neutralize the Toxins of B. jararaca Venom and Inhibit its Lethality

Abstract

The venom of the Brazilian pit viper Bothrops jararaca (BjV) is a complex mixture of molecules, and snake venom metalloproteinases (SVMP) and serine proteinases (SVSP) are the most abundant protein families found therein. Toxins present in BjV trigger most of the deleterious disturbances in hemostasis observed in snakebites, i.e., thrombocytopenia, hypofibrinogenemia and bleedings. The treatment of patients bitten by snakes still poses challenges and the bioflavonoid rutin has already been shown to improve hemostasis in an experimental model of snakebite envenomation. However, rutin is poorly soluble in water; in this study, it was succinylated to generate its water-soluble form, rutin succinate (RS), which was analyzed comparatively regarding the chemical structure and characteristic features of rutin. Biological activities of rutin and RS were compared on hemostatic parameters, and against toxic activities of crude BjV in vitro. In vivo, C57BL/6 mice were injected i.p. with either BjV alone or pre-incubated with rutin, RS or 1,10-phenanthroline (o-phe, an SVMP inhibitor), and the survival rates and hemostatic parameters were analyzed 48 h after envenomation. RS showed the characteristic activities described for rutin – i.e., antioxidant and inhibitor of protein disulfide isomerase – but also prolonged the clotting time of fibrinogen and plasma in vitro. Differently from rutin, RS inhibited typical proteolytic activities of SVMP, as well as the coagulant activity of BjV. Importantly, both rutin and RS completely abrogated the lethal activity of BjV, in the same degree as o-phe. BjV induced hemorrhages, falls in RBC counts, thrombocytopenia and hypofibrinogenemia in mice. Rutin and RS also improved the recovery of platelet counts and fibrinogen levels, and the development of hemorrhages was totally blocked in mice injected with BjV incubated with RS. In conclusion, RS has anticoagulant properties and is a novel SVMP inhibitor. Rutin and RS showed different mechanisms of action on hemostasis. Only RS inhibited directly BjV biological activities, even though both flavonoids neutralized B. jararaca toxicity in vivo. Our results showed clearly that rutin and RS show a great potential to be used as therapeutic compounds for snakebite envenomation.

[Retracted] Chemical Structures and Pharmacological Properties of Typical Bioflavonoids in Polygonati Rhizoma (PGR)

“…Artificially, rutin can be modified by adding succinate moieties on glycoside hydroxyl groups of rutin (Figure 1). Alternatively, metal ions can be conjugated to rutin [14].…”

“…Polyphenols, therefore, are a rightful candidate for snake venom treatment since they exhibit antioxidant properties. Research was done by Sachetto et al where rutin succinate was tested for antivenom properties [14].…”

“…Sachetto et al analyzed the effect of VV on prothrombin activation. The team preincubated VV with rutin succinate and found that prothrombin activation was reduced by 98%, which greatly reduced the probability and degree of coagulation and also had a large positive effect on the treatment of snake venom [14]. Rutin succinate is expected to be a natural drug replacing specific inhibitors.…”

“…After 48 h, the survival rate of 3LD 50 alone group was 33.3%. It then follows that rutin succinate prohibits lethality of mice following VV administration [14].…”

Secondary Metabolites with Biomedical Applications from Plants of the Sarraceniaceae Family

“…Another interesting element is the ability of rutin and its derivative rutin succinate to inhibit metalloproteinases from the venom of the Brazilian pit viper Bothrops jararaca , protecting mice from venom toxicity and ensuring their survival. These findings indicate that rutin has potential as complementary treatment for snakebites [ 157 , 158 ].…”