Antitrypanosomal Activity of Anthriscus Nemorosa Essential Oils and Combinations of Their Main Constituents
Abstract
:1. Introduction
2. Results
2.1. EO Compositions
2.2. Antitrypanosomal Activity
2.3. Contribution of Individual Compounds to the Overall Antitrypanosomal Activity and Synergistic Effects
2.4. Preparation and Characterization of A. nemorosa EO-Based Nanoemulsions
2.5. Determination of NTP and ADP Pools in T. brucei
3. Discussion
4. Materials and Methods
4.1. Plant Material
4.2. Chemicals
4.3. Hydrodistillation
4.4. GC-MS Analysis
4.5. T. brucei and Mammalian Cell Culture
4.6. Growth Inhibition Assay on T. brucei and Balb/3T3 Cells
4.7. Comparative Activities
4.8. Synergistic Interactions among Four Major Constituents
4.9. Preparation of Anthriscus nemorosa EO-Based Nanoemulsions
4.10. Nanoemulsion Characterization
4.11. Determination of NTP and ADP Pools in T. brucei
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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N° | Component a | RI b | RI Lit. c | % A. nemorosa EO-AP | % A. nemorosa EO-R | ID d |
---|---|---|---|---|---|---|
1 | n-nonane | 901 | 900 | 1.7 | 9.5 | RI, MS |
2 | α-thujene | 921 | 924 | Tr e | tr | RI, MS |
3 | α-pinene | 927 | 932 | 0.4 | 1.7 | Std |
4 | sabinene | 964 | 969 | 0.4 | 0.4 | Std |
5 | β-pinene | 969 | 974 | 3.0 | 22.0 | Std |
6 | myrcene | 990 | 988 | 40.1 | 19.9 | Std |
7 | n-decane | 1001 | 1000 | 0.1 | 0.8 | RI, MS |
8 | α-phellandrene | 1004 | 1002 | 0.2 | Std | |
9 | α-terpinene | 1015 | 1014 | 0.1 | tr | Std |
10 | p-cymene | 1022 | 1020 | 2.2 | 2.8 | Std |
11 | limonene | 1024 | 1024 | 5.7 | 11.5 | Std |
12 | β-phellandrene | 1026 | 1025 | 3.0 | 2.7 | Std |
13 | (Z)-β-ocimene | 1038 | 1032 | 4.4 | 8.9 | Std |
14 | (E)-β-ocimene | 1048 | 1044 | 2.8 | 1.8 | Std |
15 | γ-terpinene | 1056 | 1055 | 11.6 | 9.2 | Std |
16 | n-undecane | 1101 | 1100 | 0.4 | 0.9 | RI, MS |
17 | allo-ocimene | 1129 | 1128 | 0.2 | 0.5 | RI, MS |
18 | β-elemene | 1384 | 1389 | 0.4 | tr | RI, MS |
19 | (E)-caryophyllene | 1387 | 1417 | 0.6 | 0.1 | Std |
20 | β-copaene | 1420 | 1430 | 0.5 | RI, MS | |
21 | (E)-β-farnesene | 1457 | 1440 | 0.7 | Std | |
22 | germacrene D | 1472 | 1484 | 10.4 | RI, MS | |
23 | 1-dodecanol | 1476 | 1469 | 0.5 | RI, MS | |
24 | epi-cubebol | 1487 | 1493 | 0.3 | RI, MS | |
25 | (E,E)-α-farnesene | 1508 | 1505 | 4.0 | RI, MS | |
26 | δ-cadinene | 1517 | 1522 | 2.3 | 0.9 | RI, MS |
27 | elemol | 1543 | 1534 | 0.5 | RI, MS | |
28 | guaiol | 1591 | 1600 | 0.8 | RI, MS | |
29 | 10-epi-γ-eudesmol | 1608 | 1622 | 0.5 | RI, MS | |
30 | epi-α-muurolol + epi-α-cadinol | 1633 | 1640/1638 | 1.0 | 0.5 | RI, MS |
31 | β-eudesmol | 1639 | 1649 | 0.2 | 0.4 | RI, MS |
32 | valerianol | 1643 | 1656 | 0.1 | 0.4 | RI, MS |
33 | α-cadinol | 1646 | 1652 | 1.7 | 0.4 | RI, MS |
34 | bulnesol | 1659 | 1670 | 0.7 | RI, MS | |
Oil yield (%, w/w) | 0.06 | 0.08 | ||||
Total identified (%) | 98.1 | 98.6 | ||||
Grouped compunds (%) | ||||||
Monoterpene hydrocarbons | 74.0 | 81.5 | ||||
Oxygenated monoterpenes | ||||||
Sesquiterpenes hydrocarbons | 18.8 | 1.0 | ||||
Oxygenated sesquiterpenes | 3.0 | 4.4 | ||||
Alkanes | 2.3 | 11.2 | ||||
Others | 0.5 |
Samples | EC50 | Selectivity Index (SI) | |
---|---|---|---|
T. brucei (s427) | Balb3T3 | ||
Essential oils | μg/mL | μg/mL | |
A. nemorosa aerial parts | 1.17 ± 0.09 | 5.48 ± 0.61 | 4.65 |
A. nemorosa roots | 2.36 ± 0.09 | 5.33 ± 0.49 | 2.25 |
Pure compounds | μg/mL(μM) | μg/mL(μM) | |
β-Pinene (1): 3% AP, 22% R | 11.4 ± 2.6 (83.7) c | >100 | >8.77 |
β-Ocimene (2): 7,2% AP, 10.7% R | 1.1 ± 0.5 (8) b | >100 | >91 |
p-Cymene (3): 2.2% AP, 2.8% R | 4.5 ± 1.0 (33) b | 28 ± 7 | 6.2 |
Limonene (4): 5.7% AP, 11.5% R | 5.6 ± 1.6 (41) a | >100 | >18 |
Myrcene (5): 40.1% AP, 19.9% R | 26.2 ± 0.44 (193) | 49.6 ± 0.64 | 1.9 |
Farnesene (6): 4.7% AP | 0.84 ± 0.04 (4.11) | 4.09 ± 0.39 | 4.85 |
γ-Terpinene (7): 11.6% AP, 9.2% R | >100 | >100 | - |
Reference drug | μg/mL(μM) | μg/mL(μM) | |
Suramin | 0.025 ± 0.001 (0.0147) | >5 | >262 |
Ratio% w/w | Observed T.b. brucei EC50 (μg/mL) | Observed Balb3T3 EC50 (μg/mL) | Selectivity Index (SI) | Expected EC50 a (μg/mL) | |||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Myrcene | β-Pinene | Limonene | β-Ocimene | p-Cymene | Farnesene | Carrier (Acetone) | Wadley b | R c | S d | ||||
Mix 1 | 63.7 | 4.7 | 9.1 | 11.4 | 3.7 | 7.4 | x | 7.25 ± 0.26 | 28.3 ± 1.06 | 3.91 | 4.09 | 0.56 | Add |
Mix 2 | x | 4.7 | 9.1 | 11.4 | 3.7 | 7.4 | 63.7 | 2.25 ± 0.18 | 9.22 ± 0.67 | 4.11 | 4.54 | 2.01 | Syn |
Mix 3 | x | x | 9.1 | 11.4 | 3.7 | 7.4 | 68.4 | 1.43 ± 0.16 | 5.14 ± 0.36 | 3.63 | 4.63 | 3.23 | Syn |
Mix 4 | x | x | x | 11.4 | 3.7 | 7.4 | 77.5 | 1.58 ± 0.14 | 5.48 ± 0.31 | 3.47 | 5.00 | 3.17 | Syn |
Mix 5 | x | x | 9.1 | 11.4 | 3.7 | x | 75.8 | 1.27 ± 0.02 | 6.98 ± 0.85 | 5.46 | 7.80 | 6.14 | Syn |
Mix 6 | x | x | 9.1 | 11.4 | x | 7.4 | 72.1 | 1.06 ± 0.04 | 3.83 ± 0.19 | 3.62 | 4.81 | 4.53 | Syn |
Mix 7 | x | x | 9.1 | x | 3.7 | 7.4 | 79.8 | 1.40 ± 0.11 | 5.77 ± 0.15 | 4.11 | 8.88 | 6.34 | Syn |
Mix 8 | 63.7 | 4.7 | x | x | x | 7.4 | 24.2 | 15.3 ± 0.33 | 29.3 ± 3.42 | 1.91 | 8.58 | 0.56 | Add |
Mix 9 | 63.7 | 4.7 | x | x | 3.7 | x | 27.9 | 49.2 ± 3.71 | 66.8 ± 3.19 | 1.36 | 27.28 | 0.55 | Add |
Mix 10 | 63.7 | 4.7 | x | 11.4 | x | x | 20.2 | 27.1 ± 0.51 | 25.6 ± 1.21 | 0.94 | 7.57 | 0.28 | Ant |
Mix 11 | 63.7 | 4.7 | 9.1 | x | x | x | 22.5 | 24.3 ± 0.45 | 60.9 ± 2.87 | 2.51 | 22.4 | 0.92 | Add |
Samples | Composition |
---|---|
Control | Polysorbate 80 at 2% (w/w) in water |
NANO A | 2% (w/w) polysorbate 80 + 6% (w/w) A. nemorosa EO from aerial parts |
NANO B | 2% (w/w) polysorbate 80 + 4% (w/w) A. nemorosa EO from aerial parts + 2% (w/w) ethyl oleate |
EC50 | Selectivity Index (SI) | ||
---|---|---|---|
T. brucei (s427) | Balb3T3 | ||
Nanoemulsions | μg/mL | μg/mL | |
Control | >40,000 | >40,000 | |
NANO A | 167.4 ± 12.8 (normalized 9.89 ± 0.797) | 1399 ± 464 | 8.35 |
NANO B | 322.3 ± 43.5 (normalized 12.7 ± 1.72) | 1745 ± 347 | 5.41 |
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Baldassarri, C.; Falappa, G.; Mazzara, E.; Acquaticci, L.; Ossoli, E.; Perinelli, D.R.; Bonacucina, G.; Dall’Acqua, S.; Cappellacci, L.; Maggi, F.; et al. Antitrypanosomal Activity of Anthriscus Nemorosa Essential Oils and Combinations of Their Main Constituents. Antibiotics 2021, 10, 1413. https://doi.org/10.3390/antibiotics10111413
Baldassarri C, Falappa G, Mazzara E, Acquaticci L, Ossoli E, Perinelli DR, Bonacucina G, Dall’Acqua S, Cappellacci L, Maggi F, et al. Antitrypanosomal Activity of Anthriscus Nemorosa Essential Oils and Combinations of Their Main Constituents. Antibiotics. 2021; 10(11):1413. https://doi.org/10.3390/antibiotics10111413
Chicago/Turabian StyleBaldassarri, Cecilia, Giulia Falappa, Eugenia Mazzara, Laura Acquaticci, Elena Ossoli, Diego Romano Perinelli, Giulia Bonacucina, Stefano Dall’Acqua, Loredana Cappellacci, Filippo Maggi, and et al. 2021. "Antitrypanosomal Activity of Anthriscus Nemorosa Essential Oils and Combinations of Their Main Constituents" Antibiotics 10, no. 11: 1413. https://doi.org/10.3390/antibiotics10111413