Background
Methods
Plant materials
Extraction and fractionation
Biological investigations
In vitro assessment of the cytotoxic activity
In vitro assessment of the anti-allergic activity
In vitro assessment of the anti-inflammatory activity
Measurement of superoxide generation
Measurement of elastase release
Statistical analysis
Phytochemical investigations
General experimental procedures
Isolation of the secondary metabolites from the bioactive fractions
Spectroscopic data of compounds 1–7
Results
In vitro assessment of the cytotoxic activity of B. rupestris and B. discolor
Cell line | BRT | BRH | BRD | BRE | BRR | BDT | BDH | BDD | BDE | doxorubicin |
---|---|---|---|---|---|---|---|---|---|---|
HepG2 | 1.28 | −2.078 | −8.02 | −8.43 | −13.74 | 1.66 | −7.43 | −16.18 | −6.62 | 91.28 ± 0.3 |
MDA-MB-231 | −15.86 | − 9.70 | −18.83 | −25.52 | − 23.30 | − 6.19 | − 6.73 | 13.17 | −20.64 | 97.69 ± 0.4 |
A549 | 0.44 | −0.81 | 7.85 | −8.37 | −0.89 | 12.59 | 8.18 | 14.99 | −6.52 | 98.05 ± 0.0 |
Sample | % viability, RBL-2H3a | % inhibition of A23187-induced β-hexosaminidase releaseb | |
---|---|---|---|
100 μg/mL | 10 μg/mL | 100 μg/mL | |
BRT | 99.0 ± 1.7 | 3.0 ± 5.2 | 25.7 ± 2.1** |
BRH | 96.7 ± 4.0 | 3.3 ± 5.8 | 39.0 ± 13.1** |
BRD | 95.3 ± 8.1 | 4.3 ± 7.5 | 19.0 ± 4.4* |
BRE | 97.7 ± 4.0 | 2.0 ± 3.5 | 7.0 ± 5.2 |
BRR | 99.0 ± 1.7 | 4.3 ± 5.1 | 3.7 ± 6.4 |
BDT | 99.0 ± 1.7 | 3.7 ± 6.4 | 16.0 ± 5.0* |
BDH | 99.7 ± 0.6 | 4.3 ± 7.5 | 30.3 ± 3.1** |
BDD | 100.0 ± 0.0 | 0.0 ± 0.0 | 44.0 ± 7.8** |
BDE | 100.0 ± 0.0 | 1.7 ± 2.9 | 0.3 ± 0.6 |
In vitro assessment of the anti-allergic activity of B. rupestris and B. discolor
In vitro assessment of the anti-inflammatory activity of B. rupestris and B. discolor
Sample | Superoxide anion generationa | Elastase releasea |
---|---|---|
IC50 (μg/mL)b | IC50 (μg/mL)b | |
BRT | 4.92 ± 1.47 | 3.82 ± 0.55 |
BRH | 5.69 ± 0.80 | 3.73 ± 1.16 |
BRD | 2.99 ± 0.73 | 1.98 ± 1.54 |
BRE | 6.25 ± 3.10 | 2.71 ± 0.79 |
BRR | 3.01 ± 1.91 | > 10c |
BDT | 4.73 ± 0.97 | 5.37 ± 1.23 |
BDH | 6.25 ± 2.18 | 6.04 ± 2.32 |
BDD | 0.78 ± 0.29 | 1.57 ± 0.84 |
BDE | 5.22 ± 1.35 | 2.95 ± 1.08 |
genistein | 0.41 ± 0.09 | 4.41 ± 1.99 |
Phytochemical investigations
Dihydrodehydrodiconiferyl alcohol 4-O-β-D-glucoside (6) | Dihydrodehydrodiconiferyl alcohol 9-O-β-D-glucoside (7) | |||
---|---|---|---|---|
δ
C
| δH (Mult, Int), J in Hz |
δ
C
| δH (Mult, Int), J in Hz | |
1 | 137.09 | 134.67 | ||
2 | 111.19 | 7.03 (d, 1H), 1.84 | 110.71 | 7.02 (d, 1H), 2.01 |
3 | 150.9 | 149.05 | ||
4 | 147.6 | 147.48 | ||
5 | 117.95 | 7.14 (d, 1H), 8.43 | 116.08 | 6.78 (d, 1H), 8.09 |
6 | 119.37 | 6.93 (dd, 1H), 8.31, 2.03 | 119.72 | 6.89 (dd, 1H), 8.28, 2 |
7 | 88.48 | 5.56 (d, 1H), 5.85 | 88.98 | 5.62 (d, 1H), 6.21 |
8 | 55.68 | 3.44 (m, 1H) | 53.28 | 3.69 (m, 1H) |
9 | 65.07 | 3.80, 3.75 (m, m, 2H) | 72.46 | 4.23, 3.79 (dd, m, 2H) |
3-OCH3 | 56.79 | 3.87 (s, 3H) | 56.44 | 3.85 (s, 3H) |
1′ | 138.37 | 136.64 | ||
2′ | 114.19 | 6.73 (s, 1H) | 114.19 | 6.75 (s, 1H) |
3′ | 145.24 | 145.21 | ||
4′ | 147.6 | 147.48 | ||
5′ | 129.58 | 129.56 | ||
6′ | 118.04 | 6.72 (s, 1H) | 118.21 | 6.80 (s, 1H) |
7′ | 32.89 | 2.63 (t, 2H) | 32.89 | 2.65 (t, 2H) |
8′ | 35.84 | 1.82 (m, 2H) | 35.82 | 1.84 (m, 2H) |
9′ | 62.22 | 3.57 (t, 2H) | 62.23 | 3.59 (t, 2H) |
3′-OCH3 | 56.71 | 3.83 (s, 3H) | 56.77 | 3.88 (s, 3H) |
1″ | 102.78 | 4.88 (covered by solvent, 1H) | 104.57 | 4.38 (d, 1H), 7.79 |
2″ | 74.90 | 3.48 (m, 1H) | 75.18 | 3.25 (m, 1H) |
3″ | 77.84 | 3.39 (m, 1H) | 78.07 | 3.31 (m, 1H) |
4″ | 71.34 | 3.39 (m, 1H) | 71.66 | 3.31 (m, 1H) |
5″ | 78.19 | 3.39 (m, 1H) | 78.26 | 3.31 (m, 1H) |
6″ | 62.51 | 3.85, 3.69 (m, 2H) | 62.81 | 3.88, 3.70 (m 2H) |
In vitro assessment of the cytotoxic activity of the isolated compounds
Cell line | Conc. (μg/mL) | % Inhibition | |
---|---|---|---|
β-amyrin acetate (1) | Scopoletin (4) | ||
HepG2 | 20 | 12.1 ± 3.4 | 11.4 ± 2.1 |
10 | 16.0 ± 0.3 | 6.5 ± 1.2 | |
5 | 20.4 ± 2.3 | 0.9 ± 0.1 | |
2.5 | 8.23 ± 0.7 | 5.9 ± 1.2 | |
MDA-MB-231 | 20 | −19.6 ± 1.2 | 8.2 ± 1.0 |
10 | −13.9 ± 1.2 | 6.0 ± 0.3 | |
5 | −5.5 ± 1.2 | 10.7 ± 9.8 | |
2.5 | 7.0 ± 0.1 | 15.5 ± 0.3 | |
A549 | 20 | 2.2 ± 0.5 | 7.9 ± 0.4 |
10 | 9.0 ± 1.0 | 14.9 ± 0.4 | |
5 | 10.8 ± 0.5 | 14.6 ± 0.5 | |
2.5 | 0.4 ± 0.7 | 14.3 ± 0.7 |
In vitro assessment of the anti-allergic activity of the isolated compounds
Sample | % viability, RBL-2H3a | % inhibition of A23187-induced β-hexosaminidase releaseb | |||
---|---|---|---|---|---|
100 μM | 500 μM | 10 μM | 100 μM | 500 μM | |
β-amyrin acetate (1) | 98.3 ± 2.9 |
c
| 0.3 ± 0.6 | 0.7 ± 1.2 |
c
|
scopoletin (4) | 96.0 ± 4.0 | 93.7 ± 6.5 | 3.7 ± 4.7 | 13.7 ± 8.0 | 23.0 ± 8.0**d |
β-sitosterol-3-O-β-D-glucoside (5) | 96.0 ± 1.0 |
c
| 5.0 ± 5.6 | 7.7 ± 3.8 |
c
|
dihydrodehydrodiconiferyl alcohol 4-O-β-D-glucoside (6) | 97.3 ± 2.5 |
e
| 7.0 ± 10.4 | 12.7 ± 7.6 |
e
|
dihydrodehydrodiconiferyl alcohol 9-O-β-D-glucoside (7) | 97.7 ± 2.1 | 95.3 ± 4.2 | 3.3 ± 4.2 | 16.3 ± 5.5* | 18.0 ± 8.7* |
In vitro assessment of the anti-inflammatory activity of the isolated compounds
Sample | Superoxide anion generationa | Elastase releasea |
---|---|---|
IC50 (μΜ)b | IC50 (μΜ)b | |
β-amyrin acetate (1) | > 10 | > 10 |
scopoletin (4) | > 10 | > 10c |
β-sitosterol-3-O-β-D-glucoside (5) | > 10 | > 10 |
dihydrodehydrodiconiferyl alcohol 4-O-β-D-glucoside (6) | > 10 | > 10 |
dihydrodehydrodiconiferyl alcohol 9-O-β-D-glucoside (7) | >1d | >1d |
genistein | 1.16 ± 0.12 | 21.51 ± 6.50 |