Anti-Inflammatory Effect of Homo- and Heterodimers of Natural Enkephalinase Inhibitors in Experimental Colitis in Mice
Abstract
:1. Introduction
2. Results and Discussion
2.1. Peptide Synthesis and Purification
2.2. Effect of Sialorphin, Opiorphin and Homo- and Heterodimers on Degradation of Met-Enkephalin by NEP
2.3. Stability in Human Plasma
2.4. Anti-Inflammatory Effect in TNBS-Induced Colitis
2.5. Histological
3. Materials and Methods
3.1. Peptide Synthesis
3.2. Peptide Purification and Analysis
3.3. Homo- and Heterodimers Synthesis
3.4. Determination of Enkephalins Degradation Rates
3.5. Stability of Sialorphin, Spinorphin and Peptide X in the Human Plasma
3.6. Animals
3.7. TNBS Model
3.8. Evaluation of Colonic Damage
3.9. Determination of Tissue Myeloperoxidase Activity
3.10. Histology
3.11. Statistics
4. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Peptide | Name | Molecular Formula | HPLC tR(min) | [M]+ Calc. | [M + H]+ Found |
---|---|---|---|---|---|
I | Sialorphin (Gln-His-Asn-Pro-Arg) | C26H42N12O8 | 5.5 * | 650.3 | 651.0 |
II | Opiorphin (Gln-Arg-Phe-Ser-Arg) | C29H48N12O8 | 6.7 * | 692.3 | 693.2 |
III | Spinorphin (Leu-Val-Val-Tyr-Pro-Trp-Thr) | C45H64N8O10 | 8.3 ** | 876.5 | 877.4 |
IV | Cys-Gln-His-Asn-Pro-Arg | C29H47N13O9S | 6.2 * | 753.3 | 754.2 |
V | Cys-Gln-Arg-Phe-Ser-Arg | C32H53N13O9S | 8.8 * | 795.3 | 796.3 |
VI | Cys-Leu-Val-Val-Tyr-Pro-Trp-Thr | C48H69N9O11S | 7.3 ** | 979.2 | 1002.4 [M + Na]+ |
VII | (Cys-Gln-His-Asn-Pro-Arg)2 | C58H92N26O18S2 | 7.6 * | 1504.6 | 1505.9 |
VIII | (Cys-Gln-Arg-Phe-Ser-Arg)2 | C64H104N26O18S2 | 9.4 * | 1588.7 | 1589.2 |
IX | (Cys-Leu-Val-Val-Tyr-Pro-Trp-Thr)2 | C96H136N18O22S2 | 9.9 ** | 1956.9 | 1957.5 |
X | Cys-Gln-His-Asn-Pro-Arg │ Cys-Leu-Val-Val-Tyr-Pro-Trp-Thr | C77H114N22O20S2 | 18.6 *** | 1731.0 | 1732.5 |
XI | Cys-Gln-Arg-Phe-Ser-Arg │ Cys-Leu-Val-Val-Tyr-Pro-Trp-Thr | C80H120N22O20S2 | 19.4 *** | 1772.8 | 1774.2 |
Peptide | Inhibitor | 1000 × k [1/min] | t1/2 [min] |
---|---|---|---|
Without inhibitor * | 25.35 ± 1.05 | 27 ± 1 | |
I | Sialorphin * (Gln-His-Asn-Pro-Arg) | 8.84 ± 0.27 ** | 78 ± 2 ** |
II | Opiorphin * (Gln-Arg-Phe-Ser-Arg) | 7.38 ± 0.20 ** | 93 ± 2 ** |
V | Cys-Gln-Arg-Phe-Ser-Arg | 3.47 ± 0.12 ** | 199 ± 7 ** |
VI | Cys-Leu-Val-Val-Tyr-Pro-Trp-Thr | 6.43 ± 0.17 ** | 107 ± 3 ** |
VII | (Cys-Gln-His-Asn-Pro-Arg)2 | 4.75 ± 0.08 ** | 145 ± 2 ** |
VIII | (Cys-Gln-Arg-Phe-Ser-Arg)2 | 1.53 ± 0.01 ** | 451 ± 2 ** |
X | Cys-Gln-His-Asn-Pro-Arg │ Cys-Leu-Val-Val-Tyr-Pro-Trp-Thr | 3.27 ± 0.10 ** | 211 ± 6 ** |
XI | Cys-Gln-Arg-Phe-Ser-Arg │ Cys-Leu-Val-Val-Tyr-Pro-Trp-Thr | 1.13 ± 0.04 ** | 612 ± 22 ** |
Sample Availability: Samples of the compounds are not available from the authors. |
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Sobocińska, M.; Salaga, M.; Fichna, J.; Kamysz, E. Anti-Inflammatory Effect of Homo- and Heterodimers of Natural Enkephalinase Inhibitors in Experimental Colitis in Mice. Molecules 2020, 25, 5820. https://doi.org/10.3390/molecules25245820
Sobocińska M, Salaga M, Fichna J, Kamysz E. Anti-Inflammatory Effect of Homo- and Heterodimers of Natural Enkephalinase Inhibitors in Experimental Colitis in Mice. Molecules. 2020; 25(24):5820. https://doi.org/10.3390/molecules25245820
Chicago/Turabian StyleSobocińska, Małgorzata, Maciej Salaga, Jakub Fichna, and Elżbieta Kamysz. 2020. "Anti-Inflammatory Effect of Homo- and Heterodimers of Natural Enkephalinase Inhibitors in Experimental Colitis in Mice" Molecules 25, no. 24: 5820. https://doi.org/10.3390/molecules25245820