The In Vitro Effects of Pentamidine Isethionate on Coagulation and Fibrinolysis
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemicals, Reagents, Enzymes, and Substrates
2.2. In vitro Effect on Clotting Times of Human Plasmas
2.3. In Vitro Effect on Fibrinolysis
2.4. Effect of Pentamidine Isethionate on Common Coagulation Pathway and Fibrinolysis Enzymes
2.5. Effect of Pentamidine Isethionate on Related Enzymes
2.6. Michaelis-Menten Kinetics for Chromogenic Substrates Hydrolysis by FXa and Plasmin in the Presence of Pentamidine Isethionate
2.7. Molecular Modeling of Pentamidine Binding to the Active Site of Human FXa and Plasmin
3. Results
3.1. Pentamidine Affects the Clotting Times of Human Plasma and the Effect Is Independent of Factors VIIa, IXa, XIa, and XIIa
3.2. Pentamidine Dose-Dependently Inhibits Plasmin-Mediated Clot Lysis
3.3. Pentamidine Isethionate Selectively Inhibits Human FXa and Plasmin in the Chromogenic Substrate Hydrolysis Assays
3.4. Michaelis-Menten Kinetics of FXa and Plasmin Inhibition by Pentamidine Isethionate
3.5. Molecular Modeling Studies of Pentamidine Interaction with FXa and Plasmin
4. Discussion and Significance
Author Contributions
Funding
Conflicts of Interest
References
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Type of Human Plasma | [Pentamidine Isethionate] (µm) to Double Clotting Time |
---|---|
APTT assay | APTTEC2× |
Normal | 27.88 b) ± 2.99 c) |
Deficient of FVII | 26.71 ± 0.50 |
Deficient of FIX | 26.32 ± 3.52 |
Deficient of FXI | 29.48 ± 3.88 |
Deficient of FXII | 27.05 ± 6.05 |
PT assay | PTEC2× |
Normal | 45.70 ± 0.85 |
TT assay | TTEC2× |
Normal | 158.17 ± 30.87 |
Enzyme | IC50 (µM) | HS | ∆Y (%) |
---|---|---|---|
Thrombin | >100 b) | NA c) | NA |
FXa | 10.4 ± 0.8 d) | 0.97 ± 0.07 | 101.6 ± 2.9 |
FXIa | >100 | NA | NA |
FXIIIa | >50 | NA | NA |
Plasmin | 8.4 ± 0.8 | 1.1 ± 0.1 | 98.0 ± 3.3 |
Neutrophil Elastase | >100 | NA | NA |
Trypsin | 4.9 ± 0.8 | 1.0 ± 0.2 | 116.8 ± 7.4 |
Chymotrypsin | >1750 | NA | NA |
Inhibitor | Spectrozyme FXa | |
Pentamidine Isethionate (µM) | KM (µM) | VMAX (mAU/min) |
0 | 305.0 ± 11.8 b) | 169.4 ± 3.6 |
5 | 266.8 ± 21.2 | 114.3 ± 4.7 |
10 | 410.3 ± 18.2 | 124.2 ± 3.3 |
20 | 364.4 ± 36.6 | 63.1 ± 3.6 |
50 | 690.2± 63.1 | 60.3 ± 3.8 |
Inhibitor | Spectrozyme PL | |
Pentamidine Isethionate (µM) | KM (µM) | VMAX (mAU/min) |
0 | 46.5 ± 5.1 | 76.2 ± 2.5 |
10 | 92.9 ± 6.2 | 78.7 ± 2.0 |
20 | 176.1 ± 4.3 | 88.4 ± 1.0 |
50 | 358.9 ± 21.3 | 92.9 ± 3.2 |
100 | 645.5 ± 68.17 | 90.9 ± 6.5 |
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Al-Horani, R.A.; Clemons, D.; Mottamal, M. The In Vitro Effects of Pentamidine Isethionate on Coagulation and Fibrinolysis. Molecules 2019, 24, 2146. https://doi.org/10.3390/molecules24112146
Al-Horani RA, Clemons D, Mottamal M. The In Vitro Effects of Pentamidine Isethionate on Coagulation and Fibrinolysis. Molecules. 2019; 24(11):2146. https://doi.org/10.3390/molecules24112146
Chicago/Turabian StyleAl-Horani, Rami A., Daytriona Clemons, and Madhusoodanan Mottamal. 2019. "The In Vitro Effects of Pentamidine Isethionate on Coagulation and Fibrinolysis" Molecules 24, no. 11: 2146. https://doi.org/10.3390/molecules24112146