Fig 1.
Lapachol (1) and analogues (2–4).
Fig 2.
Proteolytic activity of B. atrox venom, and the effect of lapachol and analogues.
Panel A shows proteolytic activity curve of B. atrox venom (1–10 μg/mL) (n = 5). Panel B shows the inhibition of B. atrox venom (10 μg/mL) by lapachol (1) and analogues (numbers 2–4) at 100 μM (n = 5). Panel C shows the inhibition of proteolytic activity by lapachol and compounds 3a, 3b, 3c and 3e in a concentration-dependent way (n = 5). One-way ANOVA Dunnett’s post-hoc test * p<0.05 vs B. atrox venom (Panel B).
Fig 3.
Collagenase activity of B. atrox venom, and the effect of lapachol and analogues.
Panel A shows the collagenase activity curve of B. atrox venom (10–50 μg/mL) (n = 5). Panel B shows the inhibition of B. atrox venom (50 μg/mL) by lapachol (1) and analogues (numbers 2–4) at 100 μM (n = 5). Panel C shows the inhibition of collagenase activity by compounds 1, 3a, 3b and 3e in a concentration-dependent way (n = 5). One-way ANOVA Dunnett’s post-hoc test * p<0.05 vs B. atrox venom.
Fig 4.
Hemorrhagic activity of Bothrops venoms in mouse skin: Effect of analogue 3a.
Data demonstrates skin hemorrhage after intradermic injection of Bothrops venom in the presence of analogue 3a at 1 and 3 mg/kg (for B. atrox, panel A) and 1 and 10 mg/kg (for B. jararaca, panel B) (n = 8). One-way ANOVA Dunnett’s post-hoc test * p<0.05 vs B. atrox venom.
Fig 5.
Effect of analogue 3a on B. atrox venom edematogenic activity.
Panels A and B show edema variation after injection of PSS, B. atrox venom (1 mg/kg) alone or associated with analogue 3a (3 and 10 mg/kg), and resulting areas under the curves, respectively (n = 5). Panel C shows edema when incubated with lapachol (1) and analogues 3a and 3b at 30 mg/kg, with the respective areas under the curves (Panel D). (n = 5). Two-Way ANOVA Bonferroni’s post-hoc test * p<0.05 vs B. atrox venom (Panels A and C). One-way ANOVA Dunnett’s post-hoc test * p<0.05 vs B. atrox venom (Panels B and D).
Fig 6.
Effect of lapachol and analogues on B. atrox venom phospholipase A2 activity.
Data show the PLA2 activity of B. atrox venom alone (10 μg/mL) (considered as 100% activity) or preincubated with lapachol (1) and analogues (numbers 2–4) at 100 μM (n = 6). One-way ANOVA Dunnett’s post-hoc test *p < 0.05 vs B. atrox venom.
Fig 7.
Effect of analogue 3a on B. atrox venom myotoxicity in vitro, and lapachol and its analogues 3a and 3b in vivo.
Panel A shows EDL muscle superfused with B. atrox venom (25 μ/mL) alone or associated with analogue 3a (300 μM) (n = 4). Two-Way ANOVA Bonferroni’s post-hoc test. Panel B shows plasma CK 2 h after i.m. injection (0.1 mL) of B. atrox venom (V) alone or associated with lapachol (1), analogue 3a or 3b (30 mg/kg) (n = 5). One-way ANOVA Dunnett’s post-hoc test.
Fig 8.
Effect of analogue 3a on B. atrox venom procoagulant activity and clotting time.
Data show the time spent for mice blood collected in a capillary previously containing venom alone or associated with analogue 3a to clot (n = 8–11). One-way ANOVA Dunnett’s post-hoc test.