Synergistic Effects of Artesunate in Combination with Amphotericin B and Miltefosine against Leishmania infantum: Potential for Dose Reduction and Enhanced Therapeutic Strategies
Abstract
:1. Introduction
2. Results
2.1. Antileishmanial Activity of AS, AmB, MF, and PM against L. infantum
2.2. Cytotoxicity of AS, AmB, MF, and PM to THP-1-Derived Macrophages
2.3. Combination Effects of Artesunate and Three Drugs against Intracellular Amastigotes
2.4. Cytotoxicity of AS–AmB, AS–MF, and AS–PM Combinations to THP-1-Derived Macrophages
3. Discussion
4. Materials and Methods
4.1. Drugs
4.2. Parasite Strain
4.3. THP-1 Cell Culture
4.4. Promastigote Assay
4.5. Cytotoxicity Assay
4.6. Preparation of Promastigotes to Infect THP-1-Derived Macrophages
4.7. Intracellular Amastigote Assay and Parasite Rescue and Transformation Assay
4.8. Drug Combination Assay on Intracellular Amastigotes
4.9. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Drugs (µM) | IC50 5 (Promastigote) | IC50 (Amastigote) | CC50 6 | SI 7 (Amastigote) |
---|---|---|---|---|
AS 1 | 121.43 ± 2.05 | 5.88 ± 1.67 | 225.77 ± 21.71 | 38.40 |
AmB 2 | 0.02 ± 0.002 | 0.14 ± 0.02 | 1.20 ± 0.27 | 8.57 |
MF 3 | 2.95 ± 0.18 | 0.37 ± 0.11 | 51.12 ± 0.65 | 138.16 |
PM 4 | 114.93 ± 9.46 | 64.30 ± 2.37 | >20,000 | >311 |
Drug Combination Non-Constant Ratio (µM) 1 | Growth Inhibition (%) 2 | CI 3 | Interaction | Dose Reduction Index (DRI) 4 | |||
---|---|---|---|---|---|---|---|
Combinations | AS | AmB | AS | AmB | |||
1 | 1.00 | 0.03 | 49.1 ± 3.87 ** | 0.68 | synergism | 2.04 | 5.39 |
2 | 1.00 | 0.05 | 63.36 ± 2.27 **** | 0.55 | synergism | 3.45 | 3.85 |
3 | 1.00 | 0.10 | 86.3 ± 4.35 **** | 0.44 | synergism | 11.19 | 2.84 |
4 | 1.00 | 0.20 | 87.85 ± 5.14 **** | 0.76 | slight synergism | 12.65 | 1.48 |
5 | 2.00 | 0.03 | 5.07 ± 4.39 **** | 14.06 | very strong antagonism | 0.07 | 2.26 |
6 | 2.00 | 0.05 | 62.23 ± 2.76 *** | 0.87 | slight synergism | 1.65 | 3.80 |
7 | 2.00 | 0.10 | 84.79 ± 4.40 **** | 0.57 | synergism | 4.99 | 2.74 |
8 | 2.00 | 0.20 | 89.21 ± 2.27 **** | 0.79 | moderate synergism | 7.14 | 1.54 |
9 | 4.00 | 0.03 | 30.51 ± 3.21 **** | 4.26 | strong antagonism | 0.25 | 4.26 |
10 | 4.00 | 0.05 | 81.53 ± 2.91 *** | 0.69 | synergism | 2.02 | 5.10 |
11 | 4.00 | 0.10 | 89.87 ± 2.97 **** | 0.58 | synergism | 3.81 | 3.14 |
12 | 4.00 | 0.20 | 94.47 ± 3.72 **** | 0.67 | synergism | 6.90 | 1.91 |
13 | 8.00 | 0.03 | 85.19 ± 1.59 | 0.89 | slight synergism | 1.28 | 9.20 |
14 | 8.00 | 0.05 | 84.83 ± 1.30 | 0.98 | nearly additive | 1.25 | 5.84 |
15 | 8.00 | 0.10 | 86.14 ± 3.20 | 1.08 | nearly additive | 1.38 | 2.83 |
16 | 8.00 | 0.20 | 96.83 ± 1.62 *** | 0.61 | synergism | 5.85 | 2.28 |
Drug Combination Non-Constant Ratio (µM) 1 | Growth Inhibition (%) 2 | CI 3 | Interaction | Dose Reduction Index (DRI) 4 | |||
---|---|---|---|---|---|---|---|
Combinations | AS | MF | AS | MF | |||
1 | 1.00 | 0.06 | 25.75 ± 2.64 | 1.44 | moderate antagonism | 0.96 | 2.52 |
2 | 1.00 | 0.13 | 51.15 ± 4.18 **** | 0.84 | moderate synergism | 2.35 | 2.42 |
3 | 1.00 | 0.25 | 75.99 ± 2.55 **** | 0.55 | synergism | 5.72 | 2.63 |
4 | 1.00 | 0.50 | 87.42 ± 2.33 **** | 0.54 | synergism | 10.77 | 2.22 |
5 | 2.00 | 0.06 | 57.28 ± 1.70 ** | 0.86 | moderate synergism | 1.43 | 6.19 |
6 | 2.00 | 0.13 | 71.17 ± 5.26 **** | 0.66 | synergism | 2.34 | 4.29 |
7 | 2.00 | 0.25 | 80.36 ± 5.46 **** | 0.60 | synergism | 3.52 | 3.12 |
8 | 2.00 | 0.50 | 96.36 ± 5.51 **** | 0.25 | strong synergism | 15.81 | 5.41 |
9 | 4.00 | 0.06 | 27.86 ± 3.67 **** | 4.18 | strong antagonism | 0.26 | 2.79 |
10 | 4.00 | 0.13 | 80.84 ± 4.83 ** | 0.72 | moderate synergism | 1.80 | 6.13 |
11 | 4.00 | 0.25 | 93.08 ± 4.75 **** | 0.36 | synergism | 4.58 | 6.90 |
12 | 4.00 | 0.50 | 96.25 ± 4.71 **** | 0.32 | synergism | 7.71 | 5.30 |
13 | 8.00 | 0.06 | 90.50 ± 3.06 * | 0.62 | synergism | 1.74 | 22.84 |
14 | 8.00 | 0.13 | 92.83 ± 3.13 ** | 0.53 | synergism | 2.22 | 12.93 |
15 | 8.00 | 0.25 | 98.40 ± 1.60 *** | 0.18 | strong synergism | 7.79 | 18.97 |
16 | 8.00 | 0.50 | 97.59 ± 3.07 *** | 0.32 | synergism | 5.57 | 7.18 |
Drug Combination Non-Constant Ratio (µM) 1 | Growth Inhibition (%) 2 | CI 3 | Interaction | Dose Reduction Index (DRI) 4 | |||
---|---|---|---|---|---|---|---|
Combinations | AS | PM | AS | PM | |||
1 | 1.00 | 8.75 | 4.37 ± 4.79 **** | 7.70 | strong antagonism | 0.14 | 2.63 |
2 | 1.00 | 17.50 | 12.03 ± 3.21 *** | 3.19 | strong antagonism | 0.37 | 1.96 |
3 | 1.00 | 35.00 | 32.98 ± 4.09 | 1.47 | antagonism | 1.20 | 1.56 |
4 | 1.00 | 70.00 | 46.15 ± 3.03 ** | 1.55 | antagonism | 2.00 | 0.95 |
5 | 2.00 | 8.75 | 25.93 ± 5.44 ** | 2.45 | antagonism | 0.44 | 5.52 |
6 | 2.00 | 17.50 | 31.78 ± 4.73 * | 2.07 | antagonism | 0.57 | 3.06 |
7 | 2.00 | 35.00 | 57.06 ± 3.99 ** | 1.11 | slight antagonism | 1.50 | 2.24 |
8 | 2.00 | 70.00 | 68.63 ± 3.90 **** | 1.17 | slight antagonism | 2.36 | 1.34 |
9 | 4.00 | 8.75 | 24.73 ± 5.08 **** | 4.99 | strong antagonism | 0.21 | 5.39 |
10 | 4.00 | 17.50 | 54.13 ± 3.16 * | 1.72 | antagonism | 0.67 | 4.29 |
11 | 4.00 | 35.00 | 72.23 ± 3.90 * | 1.07 | nearly additive | 1.38 | 2.86 |
12 | 4.00 | 70.00 | 78.47 ± 1.60 *** | 1.15 | slight antagonism | 1.89 | 1.61 |
13 | 8.00 | 8.75 | 59.97 ± 3.84 **** | 2.50 | antagonism | 0.42 | 9.36 |
14 | 8.00 | 17.50 | 78.12 ± 3.54 | 1.24 | moderate antagonism | 0.93 | 6.41 |
15 | 8.00 | 35.00 | 75.94 ± 1.28 | 1.54 | antagonism | 0.83 | 3.07 |
16 | 8.00 | 70.00 | 74.88 ± 1.13 | 1.94 | antagonism | 0.78 | 1.50 |
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Intakhan, N.; Saeung, A.; Rodrigues Oliveira, S.M.; Pereira, M.d.L.; Chanmol, W. Synergistic Effects of Artesunate in Combination with Amphotericin B and Miltefosine against Leishmania infantum: Potential for Dose Reduction and Enhanced Therapeutic Strategies. Antibiotics 2024, 13, 806. https://doi.org/10.3390/antibiotics13090806
Intakhan N, Saeung A, Rodrigues Oliveira SM, Pereira MdL, Chanmol W. Synergistic Effects of Artesunate in Combination with Amphotericin B and Miltefosine against Leishmania infantum: Potential for Dose Reduction and Enhanced Therapeutic Strategies. Antibiotics. 2024; 13(9):806. https://doi.org/10.3390/antibiotics13090806
Chicago/Turabian StyleIntakhan, Nuchpicha, Atiporn Saeung, Sonia M. Rodrigues Oliveira, Maria de Lourdes Pereira, and Wetpisit Chanmol. 2024. "Synergistic Effects of Artesunate in Combination with Amphotericin B and Miltefosine against Leishmania infantum: Potential for Dose Reduction and Enhanced Therapeutic Strategies" Antibiotics 13, no. 9: 806. https://doi.org/10.3390/antibiotics13090806