Evidence of Pyrimethamine and Cycloguanil Analogues as Dual Inhibitors of Trypanosoma brucei Pteridine Reductase and Dihydrofolate Reductase
Abstract
:1. Introduction
2. Results
2.1. Structural Characterization of the TbPTR1:NADP(H):PYR Complex
2.2. Selection of the Small Library of CYC Derivatives
2.3. Synthesis of CYC Derivatives
2.4. Structure–Activity Relationship Studies of CYC Analogues against Parasitic and Human Enzymes
2.5. Compounds’ Selectivity between the PTR1 and DHFRs Enzymes
3. Discussion
3.1. Comparative Analysis of CYC versus PYR TbPTR1 Structure
3.2. Structural Basis for Developing Dual Inhibitors of TbPTR1 and TbDHFR
4. Materials and Methods
4.1. Synthesis of CYC Analogues
4.2. TbPTR1 and TbDHFR-TS Expression, Purification, Kinetics, and Inhibition Assays
4.3. TbPTR1 and TbDHFR-TS Crystallization
4.4. Data Collection, Structure Solution, and Refinement
4.5. Stability of Compound 6 in Solution at Different pH
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Compound Code | TbPTR1 (IC50 µM) | TbDHFR-TS (IC50 µM) | hDHFR (IC50 µM) | Selectivity IC50 TbDHFR-TS /IC50 TbPTR1 | Selectivity IC50 hDHFR /IC50 TbDHFR-TS |
---|---|---|---|---|---|
PYR | 0.090 | Ki = 0.024 [37] | Ki = 0.47 [38] | c.a. 0.27 a | 19.6 |
CYC | 31.6 | Ki = 0.256 [37] | Ki = 0.41 [38] | c.a. 0.002 a | 1.6 |
MTX | 0.793 | 0.040 | 0.014 | 0.05 | 0.035 |
1 (f) | 6.02 (±1.10) | 103 (±14) | 337 (±53) | 16.9 | 3.27 |
2 (a) | 27.5 (±4.54) | 7.26 (±2.35) | 66.7 (±18.5) | 0.26 | 9.19 |
3 (b) | 1.22 (±0.24) | 17.6 (±4.69) | 65.6 (±9.3) | 14.4 | 3.73 |
4 (e) | 4.10 (±0.75) | 89.0 (±9.9) | 284 (±77.4) | 21.7 | 3.19 |
5 (c) | 0.67 (±0.18) | 33.2 (±9.64) | 413 (±102) | 49.6 | 12.4 |
6 (d) | 0.83 (±0.07) | 18.9 (±5.17) | 62.5 (±9.1) | 22.8 | 3.31 |
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Tassone, G.; Landi, G.; Linciano, P.; Francesconi, V.; Tonelli, M.; Tagliazucchi, L.; Costi, M.P.; Mangani, S.; Pozzi, C. Evidence of Pyrimethamine and Cycloguanil Analogues as Dual Inhibitors of Trypanosoma brucei Pteridine Reductase and Dihydrofolate Reductase. Pharmaceuticals 2021, 14, 636. https://doi.org/10.3390/ph14070636
Tassone G, Landi G, Linciano P, Francesconi V, Tonelli M, Tagliazucchi L, Costi MP, Mangani S, Pozzi C. Evidence of Pyrimethamine and Cycloguanil Analogues as Dual Inhibitors of Trypanosoma brucei Pteridine Reductase and Dihydrofolate Reductase. Pharmaceuticals. 2021; 14(7):636. https://doi.org/10.3390/ph14070636
Chicago/Turabian StyleTassone, Giusy, Giacomo Landi, Pasquale Linciano, Valeria Francesconi, Michele Tonelli, Lorenzo Tagliazucchi, Maria Paola Costi, Stefano Mangani, and Cecilia Pozzi. 2021. "Evidence of Pyrimethamine and Cycloguanil Analogues as Dual Inhibitors of Trypanosoma brucei Pteridine Reductase and Dihydrofolate Reductase" Pharmaceuticals 14, no. 7: 636. https://doi.org/10.3390/ph14070636