Background
Methods
Animal
Plant collection and extraction
Collection and isolation of major compounds
Preparation of aglycone derivatives
Preparation of acetylated derivatives
Pharmacological tests
Spontaneous pain induced by formalin in mice
Mechanical hyperalgesia and inflammation induced by formalin in rats
Biochemical analysis
Statistical analysis
Results
Cycloartanes
Positions | AOP1
| AOP2
| AOP3
| |||
---|---|---|---|---|---|---|
δC
| δH
(mult, J(Hz)
| δC
| δH (mult, J(Hz) | δC
| δH (mult, J (Hz)
| |
1 | 31.5 | 1.33 (1H, m, Hβ), 1.61 (1H, m, Hα) | 31.3 | 1.33 (1H, m, Hβ), 1.60 (1H, m, Hα) | 31.3 | 1.33 (1H, m, Hβ), 1.62 (1H, m, Hα) |
2 | 30.5 | 1.64 (1H, m, Hα), 1.73 (1H, m, Hβ) | 29.4 | 1.64 (1H, m, Hα), 1.73 (1H, m, Hβ) | 29.4 | 1.63 (1H, m, Hα), 1.72 (1H, m, Hβ) |
3 | 79.1 | 3.17 (dd, 4.6, 10.1) | 77.7 | 3.24 (dd, 4.4, 11.1) | 78.0 | 3.17 (dd, 4.5, 10.1) |
4 | 41.2 | 39.8 | / | 39.8 | / | |
5 | 45.6 | 1.51 (dd, 3.7, 13.1) | 45.7 | 1.52 (dd, 3.9, 13.1) | 45.7 | 1.51 (dd, 3.8, 12.1) |
6 | 29.3 | 1.04 (1H, m, Hα), 1.74 (1H, m, Hβ) | 30.7 | 1.04 (1H, m, Hα), 1.74 (1H, m, Hβ) | 30.7 | 1.03, (1H, m, Hα), 1.74 (1H, m, Hβ) |
7 | 70.1 | 3.50 ddd (4,0, 7.1, 12.6) | 70.0 | 3.50 ddd (4,0, 7.1, 12.7) | 70.0 | 3.50 (ddd, 4.0, 9.4, 13.3) |
8 | 54.4 | 1.72 (1H, d, 7.0) | 54 .2 | 1.71 (1H, d, 6.0) | 54.2 | 1.72 (1H, d, 6.0) |
9 | 26.6 | 26.5 | / | 26.5 | / | |
10 | 19.7 | 19.6 | / | 19.7 | / | |
11 | 26.0 | 1.31 (1H, m, Hα), 1.90 (1H, m, Hβ) | 26.0 | 1.31 (1H, m, Hα), 1.92 (1H, m, Hβ) | 26.0 | 1.30 (1H, m, Hα), 1.92 (1H, m, Hβ) |
12 | 32.8 | 1.61 (1H, m, Hα), 1.68 (1H, m, Hβ) | 32.4 | 1.63 (1H, m, Hα), 1.69 (1H, m, Hβ) | 32.3 | 1.62 (1H, m, Hα), 1.68 (1H, m, Hβ) |
13 | 45.3 | 45.4 | / | 45.4 | / | |
14 | 46.0 | 46.0 | / | 46.0 | / | |
15 | 48.8 | 1.64 (1H, dd, 5.2, 7.5, Hβ), 2.25 (1H, dd, 5.2 7.5, Hα) | 48.8 | 1.66 (1H, dd, 5.2, 7.6, Hβ), 2.25 (1H, dd, 5.2, 7.6, Hα) | 48.8 | 1.67 (1H, dd, 5.2, 7.7, Hβ), 2.25 (1H, dd, 5.2, 7.7, Hα) |
16 | 72.1 | 4.43 (1H, ddd, 5.2, 7.5, 7.8) | 72.1 | 4.44 (1H, ddd, 5.2, 7.6, 7.9) | 72.0 | 4.43 (1H, ddd, 5.2, 7.6, 7.9) |
17 | 55.8 | 1.63 (1H, dd, 7.8, 12.3) | 55.9 | 1.64 (1H, dd, 7.9, 12.4) | 55.8 | 1.63 (1H, dd, 7.9, 12.4) |
18 | 17.2 | 1.20 (3H, s) | 17.3 | 1.22 (3H, s) | 17.2 | 1.21 (3H, s) |
19 | 27.9 | 0.19 (1H, d, 4.0, Hα), 0.87 (1H, d, 4.0, Hβ) | 27.9 | 0.19 (1H, d, 4.0, Hα), 0.85 (1H, d, 4.0, Hβ) | 27.9 | 0.75 (1H, d, 4.0, Hα), 0.86 (1H, d, 4.0, Hβ) |
20 | 30.1 | 1.81 (1H, m) | 28.4 | 1.92 (1H, m) | 28.5 | 1.82 (1H, m) |
21 | 17.5 | 0.99 (1H, d, 7.6) | 17.6 | 0.99 (1H, d, 7.6) | 17.5 | 1.00 (1H, d, 7.6) |
22 | 31.1 | 1.26 (1H, m, Hα),, 1.82 (1H, m, Hβ) | 32.3 | 1.24 (1H, m, Hα), 1.82 (1H, m, Hβ) | 32.5 | 1.22(1H, m, Hα),, 1.92(1H, m, Hβ) |
23 | 28.3 | 1.63 (1H, m, Hα), 1.70 (1H, m, Hβ) | 27.0 | 1.63 (1H, m, Hα), 1.72 (1H, m, Hβ) | 28.0 | 1.62 (1H, m, Hα), 1.72 (1H, m, Hβ) |
24 | 218.9 | 77.0 | 3.40 (1H, dd, 5.1, 12.0) | 79.3 | 3.25 (1H, dd, 4.9, 11..3) | |
25 | 77.8 | 72.4 | / | 72.4 | / | |
26 | 26.8 | 1.19 (3H, s) | 24.0 | 1.19 (3H, s) | 24.3 | 1.19 (3H, s) |
27 | 26.8 | 1.17 (3H, s) | 23.9 | 1.17 (3H, s) | 23.4 | 1.16 (3H, s) |
28 | 26.0 | 0.98 (3H, s) | 24.6 | 0.98 (3H, s) | 24.6 | 0.98 (3H, s) |
29 | 13.2 | 0.86 (3H, s) | 13.1 | 0.84 (3H, s) | 14.9 | 0.86 (3H, s) |
30 | 18.5 | 0.97 (3H, s) | 18.4 | 0.97 (3H, s) | 18.4 | 0.97 (3H, s) |
Positions | HOP1
| HOP2
| HOP3
| |||
---|---|---|---|---|---|---|
δC
| δH
(mult, J(Hz)
| δC
| δH (mult, J(Hz) | δC
| δH (mult, J (Hz)
| |
1 | 30.4 | 1.40 (1H, m, Hβ), 1.60 (1H, m, Hα) | 29.1 | 1.62 (1H, m, Hβ), 1.72 (1H, m, Hα) | 29.1 | 1.62 (1H, m, Hβ), 1.72 (1H, m, Hα) |
2 | 29.1 | 1.62 (1H, m, Hα), 1.73 (1H, m, Hβ) | 27.0 | 1.43 (1H, m, Hα), 1.62 (1H, m, Hβ) | 27.0 | 1.43 (1H, m, Hα), 1.62 (1H, m, Hβ) |
3 | 77.8 | 3.22 (dd, 4.4, 11.1) | 77.8 | 3.23 (dd, 4.2, 10.9) | 77.8 | 3.23 (dd, 4.2, 10.9) |
4 | 39.8 | / | 39.8 | / | 39.8 | / |
5 | 44.3 | 1.42 (dd, 3.9, 13.3) | 44.2 | 1.45 (dd, 3.9, 13.1) | 44.2 | 1.45 (dd, 3.9, 13.1) |
6 | 28.1 | 1.08 (1H, m, Hα), 2.13 (1H, m, Hβ) | 28.1 | 1.10 (1H, m, Hα), 2.13 (1H, m, Hβ) | 28.1 | 1.10 (1H, m, Hα), 2.13 (1H, m, Hβ) |
7 | 80.0 | 3.63 (ddd (4,0, 7.63, 12.6) | 80.0 | 3.63 (ddd (4,0, 7.1, 12.7) | 80.0 | 3.63 (ddd (4,0, 7.1, 12.7) |
8 | 50.2 | 2.09 (1H, d, 7.63) | 50.0 | 2.11 (1H, d, 6.0) | 50.0 | 2.11 (1H, d, 6.0) |
9 | 20.1 | / | 20.2 | / | 20.2 | / |
10 | 26.4 | / | 26.4 | / | 26.4 | / |
11 | 26.5 | 1.33 (1H, m, Hα), 1.90 (1H, m, Hβ) | 30.3 | 1.42 (1H, m, Hα), 1.62 (1H, m, Hβ) | 30.3 | 1.42 (1H, m, Hα), 1.62 (1H, m, Hβ) |
12 | 32.3 | 1.60 (1H, m, Hα), 1.68 (1H, m, Hβ) | 32.4 | 1.60 (1H, m, Hα), 1.69 (1H, m, Hβ) | 32.4 | 1.60 (1H, m, Hα), 1.69 (1H, m, Hβ) |
13 | 46.4 | / | 46.4 | / | 46.4 | / |
14 | 45.3 | / | 45.3 | / | 45.3 | / |
15 | 46.6 | 1.64 (1H, dd, 5.0, 7.5, Hβ), 2.33 (1H, dd, 5.0 7.5, Hα) | 46.4 | 1.63 (1H, dd, 5.2, 7.6, Hβ), 2.31 (1H, dd, 5.2, 7.6, Hα) | 46.4 | 1.63 (1H, dd, 5.2, 7.6, Hβ), 2.31 (1H, dd, 5.2, 7.6, Hα) |
16 | 71.9 | 4.46 (1H, ddd, 5.0, 7.6, 7.8) | 72.3 | 4.43 (1H, ddd, 5.2, 7.6, 7.9) | 72.3 | 4.43 (1H, ddd, 5.2, 7.6, 7.9) |
17 | 56.0 | 1.62 (1H, dd, 7.7, 12.2) | 56.0 | 1.64 (1H, dd, 7.9, 12.4) | 56.0 | 1.64 (1H, dd, 7.9, 12.4) |
18 | 16.2 | 1.18 (3H, s) | 16.1 | 1.20 (3H, s) | 16.1 | 1.20 (3H, s) |
19 | 24.9 | 0.20 (1H, d, 4.5, Hα) 0.85 (1H, d, 4.5, Hβ) | 24.7 | 0.18 (1H, d, 4.6, Hα), 0.87 (1H, d, 4.6, Hβ) | 24.7 | 0.18 (1H, d, 4.6, Hα), 0.87 (1H, d, 4.6, Hβ) |
20 | 29.7 | 1.72 (1H, m) | 28.5 | 1.95 (1H, m) | 28.5 | 1.91 (1H, m) |
21 | 17.0 | 0.97 (1H, d, 7.6) | 17.2 | 0.97 (1H, d, 7.6) | 17.2 | 0.97 (1H, d, 7.6) |
22 | 32.9 | 2.78 (1H, m, Hα), 2.86 (1H, m, Hβ) | 32.4 | 1.22 (1H, m, Hα), 1.81 (1H, m, Hβ) | 32.4 | 1.82 (1H, m, Hα), 1.01 (1H, m, Hβ) |
23 | 29.5 | 1.21 (1H, m, Hα), 1.97 (1H, m, Hβ) | 30.3 | 1.42 (1H, m, Hα), 1.62 (1H, m, Hβ) | 31.3 | 2.12 (1H, m, Hα), 1.12 (1H, m, Hβ) |
24 | 218.1 | / | 77.0 | 3.36 (1H, dd, 5.1, 12.0) | 79.5 | 3.46 (1H, dd, 5.1, 12.0) |
25 | 76.5 | / | 71.9 | / | 71.9 | / |
26 | 25.4 | 1.32 (3H, s) | 25.4 | 1.32 (3H, s) | 25.4 | 1.32 (3H, s) |
27 | 24.8 | 1.00 (3H, s) | 24.0 | 1.18 (3H, s) | 24.5 | 1.18 (3H, s) |
28 | 24.9 | 1.00 (3H, s) | 24.9 | 1.00 (3H, s) | 24.9 | 1.00 (3H, s) |
29 | 12.2 | 0.82 (3H, s) | 12.8 | 0.82 (3H, s) | 12.8 | 0.82 (3H, s) |
30 | 18.5 | 0.96 (3H, s) | 18.5 | 0.94 (3H, s) | 18.5 | 0.94 (3H, s) |
1’ | 102.6 | 4.36 (1H, d, 7.8) | 102.6 | 4.35 (1H, d, 7.7) | 102.5 | 4.35 (1H, d, 7.7) |
2’ | 73.9 | 3.18 (1H, dd, 7.8, 8.8) | 73.9 | 3.18 (1H, dd, 7.7, 8.7) | 73.9 | 3.18 (1H, dd, 7.7, 8.7) |
3’ | 76.9 | 3.37 (1H, dd, 8.5, 8.8) | 76.9 | 3.40 (1H, dd, 8.7, 9.0) | 76.9 | 3.40 (1H, dd, 8.7, 9.0) |
4’ | 70.2 | 3.28 (1H, dd, 8.5, 9.0) | 70.2 | 3.27 (1H, dd, 9.0, 9.2) | 70.2 | 3.27 (1H, dd, 9.0, 9.2) |
5’ | 73.7 | 3.48 (1H, m) | 73.7 | 3.48 (1H, m) | 73.7 | 3.48 (1H, m) |
6’ | 63.9 | 4.11 (1H, dd, 2.1, 11.9), 4.48 (1H, dd, 2.1, 11.9) | 63.9 | 4.11 (1H, dd, 2.0, 12.0), 4.48 (1H, dd, 2.0, 12) | 63.9 | 4.11 (1H, dd, 2.0, 12.0), 4.48 (1H, dd, 2.0, 12) |
Effect of the mixture and isolated cycloartanes on formalin-induced pain in mice
Effect of OP3, its aglycone (AOP1) and acetylated (HOP1) derivatives on formalin-induced pain in mice
Effect of OP5, its aglycone (AOP2) and acetylated (HOP2) derivatives on formalin-induced pain in mice
Effect of OP6, its aglycone (AOP3) and acetylated (HOP3) derivatives on formalin-induced pain in mice
Effect of OP5 and OP6 on mechanical hyperalgesia and inflammation induced by formalin in rats
Biochemical analyses
Parameters | Tissue | Control | Diclofenac (5 mg/kg) | OP5 (2.5 mg/kg) | OP5 (5 mg/kg) |
---|---|---|---|---|---|
NO (μmol/g tissue) | liver | 0.49 ± 0.06 | 0.28 ± 0.06* | 0.48 ± 0.04 | 0.36 ± 0.03 |
kidney | 1.014 ± 0.11 | 0.66 ± 0.15 | 1.23 ± 0.15 | 0.85 ± 0.06 | |
Glutathione (μmol/g tissue) | liver | 0.12 ± 0.01 | 0.11 ± 0.01 | 0.12 ± 0.01 | 0.10 ± 0.01 |
kidney | 0.07 ± 0.01 | 0.04 ± 0.01 | 0.09 ± 0.01 | 0.07 ± 0.00 | |
MDA (μmol/g tissue) | liver | 9.37 ± 0.74 | 9.80 ± 0.46 | 3.10 ± 0.37*** | 9.42 ± 0.24 |
kidney | 0.12 ± 0.01 | 0.09 ± 0.02 | 0.14 ± 0.01 | 0.12 ± 0.00 | |
SOD (U/mg protein) | liver | 0.05 ± 0.01 | 0.12 ± 0.00* | 0.08 ± 0.02 | 0.11 ± 0.01* |
kidney | 0.19 ± 0.08 | 0.12 ± 0.05 | 0.09 ± 0.02 | 0.10 ± 0.04 | |
Catalase (U/mg protein) | kidney | 0.08 ± 0.01 | 0.08 ± 0.01 | 0.10 ± 0.02 | 0.08 ± 0.01 |
ALAT (U/mg protein) | serum | 0.39 ± 0.15 | 0.4 ± 0.01 | 0.60 ± 0.08 | 0.80 ± 0.21 |
ASAT (U/mg protein) | serum | 1.28 ± 0.10 | 1.58 ± 0.13 | 1.35 ± 0.07 | 1.313 ± 0.11 |
Ulcerogenic activity
Groups | Doses (mg/kg) | Mucus weight (mg) | Total surface of stomach (mm2) | ulcerated surface (mm2) | % of ulcerated animals |
---|---|---|---|---|---|
Control | 0.163 ± 0.013 | 1479.00 ± 72.79 | 0.00 ± 0.00 | 0 | |
Diclo | 5 | 0.073 ± 0.014* | 1406.00 ± 78.92 | 221.10 ± 0.00*** | 100 |
OP5 | 2,5 | 0.127 ± 0.018 | 1886.00 ± 127.00 | 0.00 ± 0.00 | 0 |
OP5 | 5 | 0.090 ± 0.009 | 1723.00 ± 46.90 | 0.00 ± 0.00 | 0 |