Fig 1.
Synthetic route for hydrazone derivatives H1-H5.
Fig 2.
(A) Effect of H1 (20 and 40 mg/kg, p.o.), morphine (10 mg/kg, i.p.) and indomethacin (20 mg/kg, i.p.); (B) Effect of H2 (20 and 40 mg/kg, p.o.), morphine (10 mg/kg, i.p.), and indomethacin (20 mg/kg, i.p.); (C) Effect of H3 (20 and 40 mg/kg, p.o.), morphine (10 mg/kg, i.p.) and indomethacin (20 mg/kg, i.p.); (D) Effect of H4 (20 and 40 mg/kg, p.o.), morphine (10 mg/kg, i.p.) and indomethacin (20 mg/kg, i.p.); (E) Effect of H5 (20 and 40 mg/kg, p.o.), morphine (10 mg/kg, i.p.) and indomethacin (20 mg/kg, i.p.) in the acetic acid-induced writhing test in mice (n = 6, per group). Values are expressed as the mean ± SEM, where a indicates p < 0.05, significantly different from the control group, according to ANOVA, followed by Tukey’s test.
Fig 3.
(A) Effect of H1 (20 and 40 mg/kg, p.o.), morphine (10 mg/kg, i.p.) and indomethacin (20 mg/kg, i.p.); (B) Effect of H2 (20 and 40 mg/kg, p.o.), morphine (10 mg/kg, i.p.) and indomethacin (20 mg/kg, i.p.); (C) Effect of H3 (20 and 40 mg/kg, p.o.), morphine (10 mg/kg, i.p.) and indomethacin (20 mg/kg, i.p.); (D) Effect of H4 (20 and 40 mg/kg, p.o.), morphine (10 mg/kg, i.p.) and indomethacin (20 mg/kg, i.p.); (E) Effect of H5 (20 and 40 mg/kg, p.o.), morphine (10 mg/kg, i.p.) and indomethacin (20 mg/kg, i.p.) in the first phase of the formalin-induced nociception test in mice (n = 6, per group). Values are expressed as the mean ± SEM, where a indicates p < 0.05, significantly different from the control group, followed by Tukey’s test.
Fig 4.
(A) Effect of H1 (20 and 40 mg/kg, p.o.), morphine (10 mg/kg, i.p.) and indomethacin (20 mg/kg, i.p.); (B) Effect of H2 (20 and 40 mg/kg, p.o.), morphine (10 mg/kg, i.p.) and indomethacin (20 mg/kg, i.p.); (C) Effect of H3 (20 and 40 mg/kg, p.o.), morphine (10 mg/kg, i.p.) and indomethacin (20 mg/kg, i.p.); (D) Effect of H4 (20 and 40 mg/kg, p.o.), morphine (10 mg/kg, i.p.) and indomethacin (20 mg/kg, i.p.); (E) Effect of H5 (20 and 40 mg/kg, p.o.), morphine (10 mg/kg, i.p.) and indomethacin (20 mg/kg, i.p.) in the second phase of the formalin-induced nociception test in mice (n = 6, per group). Values are expressed as the mean ± SEM, where a indicates p < 0.05, significantly different from the control group, b indicates p < 0.05 in comparison with indomethacin group and c indicates p<0.05 in comparison with morphine group, according to ANOVA, followed by Tukey’s test.
Fig 5.
Effect of H5 (20 mg/kg, p.o.), naloxone (1.5 mg/kg, i.p.), naloxone + H5 and morphine (10 mg/kg, i.p.) in the first (A) and second (B) phases of the formalin-induced nociception test in mice (n = 6, per group). Values are expressed as the mean ± S.E.M., where a indicates p < 0.05 in comparison with control group, b indicates p<0.05 in comparison with morphine group, and c indicates p < 0.05 in comparison with H5 group, according to ANOVA, followed by Tukey’s post-test.
Fig 6.
Effect of L-NAME (20 mg/kg, i.p.), H5 (20 mg/kg, p.o.), L-NAME + H5, in the first (A) and second (B) phases of the formalin-induced nociception test in mice (n = 6, per group). Values are expressed as the mean ± S.E.M., where a indicates p < 0.05 in comparison with control group, according to ANOVA, followed by Tukey’s post-test.
Fig 7.
Effect of ondansetron (0.5 mg/kg, i.p.), H5 (20 mg/kg, p.o.), ondansetron + H5 in the first (A) and second (B) phases of the formalin-induced nociception test in mice (n = 6, per group). Values are expressed as the mean ± S.E.M., where a indicates p < 0.05 in comparison with control group and b indicates p < 0.05 in comparison with the ondansetron group, according to ANOVA, followed by Tukey’s post-test.
Fig 8.
Effect of atropine (0.1 mg/kg, i.p.), H5 (20 mg/kg, p.o.), atropine + H5 in the first (A) and second (B) phases of the formalin-induced nociception test in mice (n = 6, per group). Values are expressed as the mean ± S.E.M., where a indicates p < 0.05 in comparison with control group, according to ANOVA, followed by Tukey’s post-test.
Fig 9.
Effect of glibenclamide (2 mg/kg, i.p.), H5 (20 mg/kg, p.o.), glibenclamide + H5 in the first (A) and second (B) phases of the formalin-induced nociception test in mice (n = 6, per group). Values are expressed as the mean ± S.E.M., where a indicates p < 0.05 in comparison with control group, according to ANOVA, followed by Tukey’s post-test.
Fig 10.
Effect of ruthenium red (3 mg/kg, i.p.), H5 (20 mg/kg, p.o.), ruthenium red + H5 in the first (A) and second (B) phases of the formalin-induced nociception test in mice (n = 6, per group). Values are expressed as the mean ± S.E.M., where a indicates p < 0.05 in comparison with control group, according to ANOVA, followed by Tukey’s post-test.
Fig 11.
Effect of H5 (20 and 40 mg/kg, p.o.) and dexamethasone (2 mg/kg, i.p.) on leukocyte migration into the peritoneal cavity induced by carrageenan in mice.
Values are expressed as the mean ± S.E.M. (n = 6, per group), where a indicates p < 0.05, significantly different from the control group, according to ANOVA, followed by Tukey’s test.
Fig 12.
Effect of H5 (20 and 40 mg/kg, p.o.) and indomethacin (20 mg/kg, i.p.) on paw edema induced by carrageenan in mice.
The sham group was treated only with saline, whereas the control group received saline and carrageenan. Values are expressed as the mean ± S.E.M. (n = 6, per group), where a indicates p < 0.05, significantly different from the control group, according to ANOVA, followed by Tukey’s post-test.
Fig 13.
Effect of H5 (20 mg/kg, p.o.) on paw edema induced by histamine in mice.
The sham group was treated only with saline, whereas the control group received saline and histamine. Values are expressed as the mean ± S.E.M. (n = 6, per group), where a indicates p < 0.05, significantly different from the control group, according to ANOVA, followed by Tukey’s post-test.
Fig 14.
Effects of H5 (20 and 40 mg/kg, p.o.), and diazepam (2.5 mg/kg, i.p.) in the Rota-rod test in mice.
Values are expressed as mean ± S.E.M. (n = 6, per group), where a indicates p < 0.05, significantly different from the control group, according to ANOVA, followed by Tukey’s post-test.
Table 1.
Lethality rate of Artemia salina nauplius to hydrazone (H5).
Table 2.
Artemia salina toxicity test of hydrazone H5.
Fig 15.
Results of the docking procedures for H5 and Meloxicam, (A) Interaction profile of H5 in the murine COX-2 enzyme binding site after the docking study; (B) Interaction profile Meloxicam in the murine COX-2 enzyme binding site after the redocking study.
Table 3.
Comparative in silico physicochemical properties and ADMET profile of the anti-inflammatory drugs indomethacin and meloxicam and new hydrazone series H1 to H5.