Novel Selective and Low-Toxic Inhibitor of LmCPB2.8ΔCTE (CPB) One Important Cysteine Protease for Leishmania Virulence
Abstract
:1. Introduction
2. Materials and Methods
2.1. Synthesis of Guanidines
2.2. Structural Characterization
2.2.1. NMR Measurements
2.2.2. Single Crystal X-ray Diffraction Analysis
2.3. Biological Toxicity Studies
2.3.1. Cell Toxicity
2.3.2. Organ Toxicity on Isolated Tissue Preparations
2.4. Inhibitory Activity on the Enzyme LmCPB2.8ΔCTE (CPB)
2.4.1. Enzyme Kinetic Assay
2.4.2. Proteases Assays
2.5. Leishmanicidal Activity
2.5.1. Parasites
2.5.2. Antileishmanial In Vivo Assay
2.5.3. Toxicity Assay for BALB/c Mice
2.5.4. Statistical Analysis
2.5.5. Ethics Statement
2.6. Docking Investigation
3. Results and Discussion
3.1. Structural Characterization
3.2. Biological Assays
3.2.1. Cell and Organ Toxicity
3.2.2. Inhibition of Leishmania Cysteine Protease
3.2.3. Leishmanicidal Activity
Reduction of Parasite Load by LQOF-G6 in BALB/c Mice Infected with L. amazonensis
Investigation of Hepatic or Renal Disturbance
3.3. Conformational Analysis
3.3.1. Investigation of Spatial Coupling by NOESY
3.3.2. Solid-state NMR
3.4. Docking Investigation
4. 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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Groups | Treatment |
---|---|
1 | Infected and treated with PBS + 1% DMSO |
2 | Infected and treated with Amp B 2 mg/kg/day |
3 | Infected and treated with LQOF-G6 2 mg/kg/day |
4 | Uninfected and untreated |
Sample | Instrument | Source | Temp. | Detector Distance | Time/Frame | #Frames | Frame Width | CCDC |
---|---|---|---|---|---|---|---|---|
[K] | [mm] | [s] | [°] | |||||
LQOF-G6 | Bruker D8 Venture | Mo | 120 | 40 | 60 | 3506 | 0.3 | 2,144,089 |
LQOF-G1 | Bruker D8 Venture | Mo | 100 | 34 | 15 | 4215 | 0.5 | 1,992,228 |
Molecule | Entry | Atoms | Angle/° (A) | Atoms | Length (A) |
---|---|---|---|---|---|
LQOF-G6 | 1 | C2A-N1A-C1A-N3A | 0.9 (3) | H(C3A)-H(N3A) | 2.90 |
2 | C1A-N1A-C2A-C3A | 81.2(3) | H(C3A)-H(N1A) | 3.07 | |
3 | C1A-N1A-C2A-C7A | −98.6(2) | H(C7A)-H(N3A) | 3.43 | |
4 | N2A-C12A-C13A-C14A | 8.3(3) | H(C7A)-H(N1A) | 2.93 | |
LQOF-G1 | 1 | C2A-N1A-C1A-N3A | 2.9 (3) | H(C3A)-H(N3A) | 4.46 |
2 | C1A-N1A-C2A-C3A | −36.4 (3) | H(C3A)-H(N1A) | 2.30 | |
3 | C1A-N1A-C2A-C7A | 14.0 (2) | H(C7A)-H(N3A) | 1.88 | |
4 | N2A-C12A-C13A-C14A | 3.7 (3) | H(C7A)-H(N1A) | 3.43 |
Ligand | Affinity/kcal.mol−1 | Inhibition Index/% |
---|---|---|
LQOF-G1 | −7.9 | - |
LQOF-G2 | −8.3 | - |
LQOF-G6 | −8.8 | 73 |
LQOF-G32 | −8.2 | 53 |
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Moreira, V.P.; da Silva Mela, M.F.; Anjos, L.R.d.; Saraiva, L.F.; Arenas Velásquez, A.M.; Kalaba, P.; Fabisiková, A.; Clementino, L.d.C.; Aufy, M.; Studenik, C.; et al. Novel Selective and Low-Toxic Inhibitor of LmCPB2.8ΔCTE (CPB) One Important Cysteine Protease for Leishmania Virulence. Biomolecules 2022, 12, 1903. https://doi.org/10.3390/biom12121903
Moreira VP, da Silva Mela MF, Anjos LRd, Saraiva LF, Arenas Velásquez AM, Kalaba P, Fabisiková A, Clementino LdC, Aufy M, Studenik C, et al. Novel Selective and Low-Toxic Inhibitor of LmCPB2.8ΔCTE (CPB) One Important Cysteine Protease for Leishmania Virulence. Biomolecules. 2022; 12(12):1903. https://doi.org/10.3390/biom12121903
Chicago/Turabian StyleMoreira, Vitor Partite, Michele Ferreira da Silva Mela, Luana Ribeiro dos Anjos, Leonardo Figueiredo Saraiva, Angela M. Arenas Velásquez, Predrag Kalaba, Anna Fabisiková, Leandro da Costa Clementino, Mohammed Aufy, Christian Studenik, and et al. 2022. "Novel Selective and Low-Toxic Inhibitor of LmCPB2.8ΔCTE (CPB) One Important Cysteine Protease for Leishmania Virulence" Biomolecules 12, no. 12: 1903. https://doi.org/10.3390/biom12121903