Polyoxygenated Steroids from the Octocoral Leptogorgia punicea and in Vitro Evaluation of Their Cytotoxic Activity
Abstract
:1. Introduction
2. Results and Discussion
2.1. Chemistry
Position | 1 | 2 | 3 | 4 | 5 |
---|---|---|---|---|---|
1 | 31.8 | 32.0 | 32.3 | 32.2 | 32.4 |
2 | 30.5 | 30.7 | 30.9 | 30.9 | 31.0 |
3 | 67.2 | 67.3 | 67.6 | 67.6 | 67.6 |
4 | 40.4 | 40.5 | 40.8 | 40.8 | 40.9 |
5 | 75.9 | 75.3 | 76.3 | 76.3 | 75.7 |
6 | 75.7 | 75.9 | 76.0 | 76.1 | 76.3 |
7 | 32.2 | 31.6 | 32.6 | 32.6 | 32.0 |
8 | 27.1 | 30.9 | 27.5 | 27.6 | 31.3 |
9 | 48.0 | 45.5 | 48.5 | 48.5 | 45.9 |
10 | 39.0 | 38.5 | 39.4 | 39.4 | 38.9 |
11 | 69.0 | 21.3 | 68.3 | 69.4 | 21.5 |
12 | 49.4 | 34.7 | 49.6 | 49.6 | 34.5 |
13 | 41.7 | 40.5 | 42.0 | 42.0 | 47.1 |
14 | 57.4 | 55.4 | 57.9 | 57.9 | 55.8 |
15 | 23.8 | 23.9 | 28.6 | 28.4 | 24.3 |
16 | 24.1 | 28.1 | 24.4 | 24.4 | 27.6 |
17 | 56.8 | 56.4 | 56.9 | 57.0 | 56.6 |
18 | 14.8 | 61.2 | 15.4 | 15.4 | 60.7 |
19 | 19.6 | 16.5 | 20.0 | 20.0 | 16.9 |
20 | 35.8 | 36.3 | 40.5 | 40.3 | 40.3 |
21 | 18.7 | 19.3 | 21.3 | 21.2 | 22.7 |
22 | 36.1 | 36.3 | 138.1 | 133.7 | 137.8 |
23 | 27.8 | 23.6 | 126.0 | 135.5 | 127.8 |
24 | 39.5 | 39.5 | 42.3 | - | 42.3 |
25 | 28.0 | 28.0 | 28.9 | 31.3 | 28.9 |
26 | 22.5 | 22.6 | 22.7 | 23.2 | 22.7 |
27 | 22.8 | 22.8 | 22.7 | 23.2 | 22.7 |
6-OAc | 170.3 | 171.3 | 170.7 | 170.7 | 170.5 |
21.5 | 21.4 | 21.9 | 21.8 | 21.8 |
Position | 1 | 2 | 3 | 4 | 5 |
---|---|---|---|---|---|
1 | 1.65 (m a) | 1.45 (m a) | 1.65 (m a) | 1.65 (m a) | 1.45 (m a) |
2α | 1.87 (m a) | 1.86 (m a) | 1.87 (m a) | 1. 87 (m a) | 1.86 (m a) |
2β | 1.58 (m a) | 1.52 (m a) | 1.57 (m a) | 1.58 (m a) | 1.54 (m a) |
3 | 4.05 (m a) | 4.09 (m a) | 4.05 (m a) | 4.06 (m a) | 4.08 (m a) |
4α | 1.54 (m a) | 1.58 (m a) | 1.54(m a) | 1.56(m a) | 1.57 (m a) |
4β | 1.95 (m a) | 1. 85 (m a) | 1.95 (m a) | 1.96 (m a) | 1.85 (m a) |
6 | 4.67 (dd, 3.2,3.2) | 4.7 (dd, 3.0,3.0) | 4.66 (dd, 3.0,3.0) | 4.66 (dd, 3.1,3.1) | 4.70 (dd, 2.8,2.8) |
7 | 1.70 (m a) | 1.61 (m a) | 1.70 (m a) | 1.70 (m a) | 1.59 (m a) |
8 | 1.96 (m a) | 1.68 (m a) | 1.95 (m a) | 1.96 (m a) | 1.67 (m a) |
9 | 1.51 (m a) | 1.38 (m a) | 1.50 (m a) | 1.51 (m a) | 1.38 (m a) |
11α | 4.19 (dd, 6.5, 3.2) | 1.48 (m a) | 4.19 (dd, 6.5, 3.2) | 4.19 (dd, 6.3, 3.3) | 1.48 (m a) |
11β | - | 1.31 (m a) | - | - | 1.30 (m a) |
12α | 1.45 (m a) | 1.04 (m a) | 1.44 (m a) | 1.46 (m a) | 1.00 (m a) |
12β | 2.15 (dd, 14.0, 2.7) | 2,46 (td, 12.7, 3.0) | 2.11 (dd, 14.0, 2.7) | 2.11 (dd, 14.2, 2.9) | 2.42 (td, 12.7, 3.0) |
14 | 1.08 (m a) | 1.24 (m a) | 1.08 (m a) | 1.07 (m a) | 1.21 (m a) |
15α | 1.32 (m a) | 1.57 (m a) | 1.66 (m a) | 1.64 (m a) | 1.55 (m a) |
15β | 1.10 (m a) | 1.01 (m a) | 1.26 (m a) | 1.25 (m a) | 0.95 (m a) |
16α | 1.59 (m a) | 1.89 (m a) | 1.55(m a) | 1.57 (m a) | 1.75 (m a) |
16β | 1.08 (m a) | 1.41 (m a) | 1.08 (m a) | 1.08 (m a) | 1.42 (m a) |
17 | 1.06 (m a) | 1.20 (m a) | 1.10 (m a) | 1.08 (m a) | 1.28 (m a) |
18a | 0.91 (s) | 3.73 (dd, 11.4, 4.7) | 0.93 (s) | 0.92 (s) | 3.62 (d, 11.7) |
18b | 3.61 (dd, 11.4, 4.7) | 3.55 (d, 11.7) | |||
19 | 1.38 (s) | 1.15 (s) | 1.38 (s) | 1.38 (s) | 1.15 (s) |
20 | 1.36 (m a) | 1.57 (m a) | 2.01 (m a) | 1.97 (m a) | 2.29 (dd, 15.3, 8.4) |
21 | 0.92 (d, 6.6) | 1.03 (d, 6.6) | 1.01 (d, 6.5) | 1.00 (d, 6.6) | 1.09 (d, 6.6) |
22 | 1.32 (m a) 0.98 (m a) | 1.37 (m a) 1.04 (m a) | 5.16 (dd, 15.0, 9.0) | 5.13 (dd, 15.0, 8.4) | 5.33 (dd, 15.0, 8.5) |
23 | 1.81 (m a) 1.25 (m a) | 1.18 (m a) 1,33 (m a) | 5.26 (dd, 15.0, 7.0) | 5.26 (dd, 15.0, 6.6) | 5.37 (dd, 15.0, 6.5) |
24 | 1.11 (m a) | 1.13 (m a) | 1.82 (m a) | - | 1.85 (m a) |
25 | 1.51 (m a) | 1.51 (m a) | 1.56 (m a) | 2.18 (m a) | 1.58 (m a) |
26 | 0.86 (d, 6.6) | 0.86 (d, 6.6) | 0.86 (d, 6.7) | 0.94 (d, 6.6) | 0.87 (d, 6.7) |
27 | 0.87 (d, 6.6) | 0.87 (d, 6.6) | 0.86(d, 6.7) | 0.94 (d, 6.6) | 0.87 (d, 6.7) |
6-OAc | 2.07 (s) | 2.06 (s) | 2.07(s) | 2.07 (s) | 2.06 (s) |
2.2. Biological Activity
Punicinol | IC50 24 h (µM) | IC50 48 h (µM) |
---|---|---|
A | 11.8 ± 1.4 | 9.6 ± 1.1 |
B | 13.6 ± 2.1 | 9.7 ± 1.7 |
C | 47.9 ± 1.5 | 35.9 ± 1.4 |
D | 81.3 ± 5.0 | 73.3 ± 0.6 |
E | 59.0 ± 2.1 | 35.8 ± 1.0 |
Cisplatin | 24.8 ± 2.1 | 16.2 ± 2.4 |
Paclitaxel | >1.0 | 0.247 ± 0.04 |
Compounds Combination Ratio | Punicinol A (µM) | Paclitaxel (µM) | Experimental CI Values | Description (Graded Symbols) |
---|---|---|---|---|
4 × IC50 | 40 | 1.000 | 1.218 | Moderate antagonism (++) |
2 × IC50 | 20 | 0.500 | 1.248 | Moderate antagonism (++) |
1 × IC50 | 10 | 0.250 | 0.519 | Synergism (+++) |
0.5 × IC50 | 5 | 0.125 | 0.485 | Synergism (+++) |
0.25 × IC50 | 2.5 | 0.063 | 0.346 | Synergism (+++) |
0.125 × IC50 | 1.25 | 0.031 | 0.180 | Strong synergism (++++) |
0.062 × IC50 | 0.625 | 0.016 | 0.116 | Strong synergism (++++) |
0.031 × IC50 | 0.31 | 0.008 | 0.095 | Very Strong synergism (+++++) |
Compounds Combination ratio | Punicinol B (µM) | Paclitaxel (µM) | Experimental CI values | Description (graded symbols) |
---|---|---|---|---|
4 × IC50 | 40 | 1.000 | 0.735 | Moderate Synergism (++) |
2 × IC50 | 20 | 0.500 | 0.633 | Synergism (+++) |
1× IC50 | 10 | 0.250 | 0.922 | Additive effect (±) |
0.5 × IC50 | 5 | 0.125 | 1.024 | Additive effect (±) |
0.25 × IC50 | 2.5 | 0.063 | 6.381 | Strong antagonism (++++) |
0.125 × IC50 | 1.25 | 0.031 | 3.191 | Antagonism (−−−) |
0.062 × IC50 | 0.625 | 0.016 | 1.595 | Antagonism (−−−) |
0.031 × IC50 | 0.31 | 0.008 | 0.799 | Synergism (+++) |
3. Experimental Section
3.1. General
3.2. Biological Material Collection
3.3. Extraction and Isolation
3.4. Biological Activity
3.4.1. Cell Culture
3.4.2. Sulforhodamine B Assay
3.4.3. Synergistic Effects of Punicinols A and B in Combination with Paclitaxel
3.4.4. Cell Cycle Analysis by Flow Cytometry
3.4.5. Clonogenic Assay
4. Conclusions
Supplementary Files
Supplementary File 1Acknowledgments
Author Contributions
Conflicts of Interest
References
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Moritz, M.I.G.; Marostica, L.L.; Bianco, É.M.; Almeida, M.T.R.; Carraro, J.L.; Cabrera, G.M.; Palermo, J.A.; Simões, C.M.O.; Schenkel, E.P. Polyoxygenated Steroids from the Octocoral Leptogorgia punicea and in Vitro Evaluation of Their Cytotoxic Activity. Mar. Drugs 2014, 12, 5864-5880. https://doi.org/10.3390/md12125864
Moritz MIG, Marostica LL, Bianco ÉM, Almeida MTR, Carraro JL, Cabrera GM, Palermo JA, Simões CMO, Schenkel EP. Polyoxygenated Steroids from the Octocoral Leptogorgia punicea and in Vitro Evaluation of Their Cytotoxic Activity. Marine Drugs. 2014; 12(12):5864-5880. https://doi.org/10.3390/md12125864
Chicago/Turabian StyleMoritz, Maria Izabel G., Lucas Lourenço Marostica, Éverson M. Bianco, Maria Tereza R. Almeida, João L. Carraro, Gabriela M. Cabrera, Jorge A. Palermo, Cláudia M. O. Simões, and Eloir P. Schenkel. 2014. "Polyoxygenated Steroids from the Octocoral Leptogorgia punicea and in Vitro Evaluation of Their Cytotoxic Activity" Marine Drugs 12, no. 12: 5864-5880. https://doi.org/10.3390/md12125864