Study of acute toxicity, endothelial- and cardioprotective properties of phenolic and thiophenolic derivatives of 2H-imidazoles

The aim. To study the acute toxicity, endothelial- and cardioprotective properties of phenolic and thiophenolic derivatives of 2H-imidazoles.

Materials and methods. The study was performed on white laboratory female BALB/c mice (n=57) and male C57Bl/6 mice (n=66). Acute toxicity was assessed according to the interstate standard GOST 32644-2014 with histological evaluation of internal organs. Endothelial dysfunction was modeled by 7-day intraperitoneal administration of N-nitro-L-arginine methyl ester (L-NAME). The studied small molecules were administered intragastrically using a probe. To assess the endothelial protective effect, the levels of systolic and diastolic blood pressure were evaluated, as well as the coefficient of endothelial dysfunction; for the cardioprotective effect, the results of stress tests on the myocardium were evaluated.

Results. The study of acute toxicity of the studied small molecules allowed us to classify them as class 4 and 5. The administration of compounds 1(a–d) and 2(a–c) to mice at a dose equal to 1/10 of LD50 led to changes in blood pressure and restoration of the dynamics of pharmacological tests in response to the administration of acetylcholine and sodium nitroprusside. Molecules 1b and 2c showed statistically significant endothelial protective activity in 3 doses (1/10, 1/50 and 1/100 of LD50). Also, these hit compounds demonstrated cardioprotective effects, recorded by the restoration of the functional capabilities of the myocardium in response to load and in the adrenoreactivity test, and to a lesser extent, during resistance exercise.

Conclusion. The studied compounds have low toxicity and have endothelial- and cardioprotective effects. This study may contribute to the formation of an idea about further directions in the study of the pharmacological activity of these molecules from the group of phenolic and thiophenolic derivatives of 2H-imidazoles.

1. Su JB. Vascular endothelial dysfunction and pharmacological treatment. World J Cardiol. 2015;7(11):719–741. DOI: 10.4330/wjc.v7.i11.719

2. Kolesnichenko PD, Shcheblykin DV, Demidenko AN, Reznikov KM, Azeez AMA, Zhuchenko MA, Demchenko SA, Shcheblykina OV. Modeling L-name induced nitric oxide deficiency considering the the cardio-and endothelial protective effects. Research Results in Pharmacology. 2019. – Vol. 11, No 4. – P. 1618–1624. DOI: 10.31838/ijpr/2019.11.04.0320

3. Strassheim D, Verin A, Batori R, Nijmeh H, Burns N, Kovacs-Kasa A, Umapathy NS, Kotamarthi J, Gokhale YS, Karoor V, Stenmark KR, Gerasimovskaya E. P2Y Purinergic Receptors, Endothelial Dysfunction, and Cardiovascular Diseases. Int J Mol Sci. 2020;21(18):6855. DOI: 10.3390/ijms21186855

4. Shcheblykin DV, Bolgov AA, Pokrovskii MV, Stepenko JV, Tsuverkalova JM, Shcheblykina OV, Golubinskaya PA, Korokina LV. Endothelial dysfunction: developmental mechanisms and therapeutic strategies. Research Results in Pharmacology. 2022;8(4):115–139. DOI: 10.3897/rrpharmacology.8.80376

5. Maujean T, Girard N, Ganesan A, Gulea M, Bonnet D. Three cheers for nitrogen: aza-DKPs, the aza analogues of 2,5-diketopiperazines. RSC Adv. 2020;10(71):43358–43370. DOI: 10.1039/d0ra09457a

6. Rani N, Singh R, Kumar P. Imidazole and Derivatives Drugs Synthesis: A Review. Curr Org Synth. 2023;20(6):630–662. DOI: 10.2174/1570179420666221118100525

7. Cornec AS, Monti L, Kovalevich J, Makani V, James MJ, Vijayendran KG, Oukoloff K, Yao Y, Lee VM, Trojanowski JQ, Smith AB 3rd, Brunden KR, Ballatore C. Multitargeted Imidazoles: Potential Therapeutic Leads for Alzheimer's and Other Neurodegenerative Diseases. J Med Chem. 2017;60(12):5120–5145. DOI: 10.1021/acs.jmedchem.7b00475

8. Ganesh KN, Zhang D, Miller SJ, Rossen K, Chirik PJ, Kozlowski MC, Zimmerman JB, Brooks BW, Savage PE, Allen DT, Voutchkova-Kostal AM. Green Chemistry: A Framework for a Sustainable Future. Environ Sci Technol. 2021;55(13):8459–8463. DOI: 10.1021/acs.est.1c03762

9. Zlotin SG, Egorova KS, Ananikov VP, et al. The green chemistry paradigm in modern organic synthesis. Russ Chem Rev. 2023;92(12):RCR5104.

10. Scioli MG, Storti G, D'Amico F, Rodríguez Guzmán R, Centofanti F, Doldo E, Céspedes Miranda EM, Orlandi A. Oxidative Stress and New Pathogenetic Mechanisms in Endothelial Dysfunction: Potential Diagnostic Biomarkers and Therapeutic Targets. J Clin Med. 2020;9(6):1995. DOI: 10.3390/jcm9061995

11. Moseev TD, Nikiforov EA, Varaksin MV, Charushin VN, Chupakhin ON. Metal-Free C-H/C-H Coupling of 2H-Imidazole 1-Oxides with Polyphenols toward Imidazole-Linked Polyphenolic Compounds. J Org Chem. 2021;86(19):13702–13710. DOI: 10.1021/acs.joc.1c01796

12. Nikiforov EA, Vaskina NF, Moseev TD, Varaksin MV, Charushin VN, Chupakhin ON. Metal-Free Eliminative C-H Arylthiolation of 2H-Imidazole N-Oxides with Thiophenols. Chemistry. 2023;5(3):1477–1487. DOI: 10.3390/chemistry5030100

13. Shtabsky BM, Grzhegotsky MI, Grzhegotsky MD, Kudrina VN, Manenko AK, Fedorenko VI. On the methodology for determining the average lethal doses and concentrations of chemicals. Sanitation and hygiene. 1980;(10):49–51. Russian

14. Medina-Leyte DJ, Zepeda-García O, Domínguez-Pérez M, González-Garrido A, Villarreal-Molina T, Jacobo-Albavera L. Endothelial Dysfunction, Inflammation and Coronary Artery Disease: Potential Biomarkers and Promising Therapeutical Approaches. Int J Mol Sci. 2021;22(8):3850. DOI: 10.3390/ijms22083850

15. Korokina LV, Golubev IV, Pokopejko ON, Zagrebelnaya AV, Demchenko SA. Search for new pharmacological targets for increasing the efficiency of correction of cardiovascular diseases. Research Results in Pharmacology. 2019;5(3):67–77. DOI: 10.3897/rrpharmacology.5.39521

16. Cyr AR, Huckaby LV, Shiva SS, Zuckerbraun BS. Nitric Oxide and Endothelial Dysfunction. Crit Care Clin. 2020;36(2):307–321. DOI: 10.1016/j.ccc.2019.12.009

17. Förstermann U, Sessa WC. Nitric oxide synthases: regulation and function. Eur Heart J. 2012;33(7):829–37, 837a–837d. DOI: 10.1093/eurheartj/ehr304

18. Lind M, Hayes A, Caprnda M, Petrovic D, Rodrigo L, Kruzliak P, Zulli A. Inducible nitric oxide synthase: Good or bad? Biomed Pharmacother. 2017;93:370–375. DOI: 10.1016/j.biopha.2017.06.036

19. Minozzo BR, Fernandes D, Beltrame FL. Phenolic Compounds as Arginase Inhibitors: New Insights Regarding Endothelial Dysfunction Treatment. Planta Med. 2018;84(5):277–295. DOI: 10.1055/s-0044-100398

20. Wang L, Bhatta A, Toque HA, Rojas M, Yao L, Xu Z, Patel C, Caldwell RB, Caldwell RW. Arginase inhibition enhances angiogenesis in endothelial cells exposed to hypoxia. Microvasc Res. 2015;98:1–8. DOI: 10.1016/j.mvr.2014.11.002

21. Kavalukas SL, Uzgare AR, Bivalacqua TJ, Barbul A. Arginase inhibition promotes wound healing in mice. Surgery. 2012;151(2):287–295. DOI: 10.1016/j.surg.2011.07.012

22. Dimitroulas T, Sandoo A, Kitas GD. Asymmetric dimethylarginine as a surrogate marker of endothelial dysfunction and cardiovascular risk in patients with systemic rheumatic diseases. Int J Mol Sci. 2012;13(10):12315–1235. DOI: 10.3390/ijms131012315

23. Lee TS, Lu TM, Chen CH, Guo BC, Hsu CP. Hyperuricemia induces endothelial dysfunction and accelerates atherosclerosis by disturbing the asymmetric dimethylarginine/dimethylarginine dimethylaminotransferase 2 pathway. Redox Biol. 2021;46:102108. DOI: 10.1016/j.redox.2021.102108

24. Kass DA, Champion HC, Beavo JA. Phosphodiesterase type 5: expanding roles in cardiovascular regulation. Circ Res. 2007;101(11):1084–1095. DOI: 10.1161/CIRCRESAHA.107.162511

25. Aversa A, Letizia C, Francomano D, Bruzziches R, Natali M, Lenzi A. A spontaneous, double-blind, double-dummy cross-over study on the effects of daily vardenafil on arterial stiffness in patients with vasculogenic erectile dysfunction. Int J Cardiol. 2012;160(3):187–191. DOI: 10.1016/j.ijcard.2011.04.003

26. Kuruppu S, Smith AI. Endothelin Converting Enzyme-1 phosphorylation and trafficking. FEBS Lett. 2012;586(16):2212–2217. DOI: 10.1016/j.febslet.2012.06.020

27. Kossmann S, Hu H, Steven S, Schönfelder T, Fraccarollo D, Mikhed Y, Brähler M, Knorr M, Brandt M, Karbach SH, Becker C, Oelze M, Bauersachs J, Widder J, Münzel T, Daiber A, Wenzel P. Inflammatory monocytes determine endothelial nitric-oxide synthase uncoupling and nitro-oxidative stress induced by angiotensin II. J Biol Chem. 2014;289(40):27540–27550. DOI: 10.1074/jbc.M114.604231

28. Watanabe T, Barker TA, Berk BC. Angiotensin II and the endothelium: diverse signals and effects. Hypertension. 2005;45(2):163–169. DOI: 10.1161/01.HYP.0000153321.13792.b9

29. Landmesser U, Dikalov S, Price SR, McCann L, Fukai T, Holland SM, Mitch WE, Harrison DG. Oxidation of tetrahydrobiopterin leads to uncoupling of endothelial cell nitric oxide synthase in hypertension. J Clin Invest. 2003;111(8):1201–1209. DOI: 10.1172/JCI14172