Synthesis and ophthalmic hypotensive effect of new potential benzimidazole-based Rho-kinase-2 inhibitors

Authors

DOI:

https://doi.org/10.18413/rrpharmacology.12.1121

Abstract

Introduction: Based on the chemical structures of known Rho-kinase-2 inhibitors, 18 benzimidazole derivatives were synthesized and studied for their ophthalmic hypotensive activity in animals with normal intraocular pressure. The dependence of the pharmacological effect on the chemical structure of the compounds was analyzed. The effect of the most active compounds on Rho-kinase-2 activity was assessed.

Materials and Methods: Ophthalmic hypotensive activity was assessed by measuring intraocular pressure with a TonoVet veterinary tonometer in 120 mongrel rats (6 in each group) before and after instillation of reference drug solutions (timolol and melatonin) and 18 test substances. The effect of the test compounds on Rho-kinase activity was assessed using an in vitro enzyme-linked immunosorbent assay spectrophotometrically.

Results: The most active compounds among the new benzimidazole derivatives after a single instillation at a concentration of 0.4% were compound 7a (1-(4-fluorobenzyl)-3-(2-(pyrrolidin-1-yl)ethyl)-1,3-dihydro-2H-benzo[d]imidazol-2-imine hydrochloride), which reduced intraocular pressure in normotensive animals by 28.21%, exceeding the effect of the reference drug timolol(26.84%), but inferior to melatonin (30.95%), and compound 1d (1-(1-(2-(azepan-1-yl)ethyl)-1H-benzo[d]imidazol-2-yl)-3-(3-trifluoromethyl)phenyl)urea hydrochloride), which reduced ophthalmotonus by 23.96%, slightly inferior to timolol. The test compounds do not affect intraocular pressure dynamics in the contralateral eye and, therefore, do not have a systemic effect, unlike the reference drugs. It was also found that compounds 7a and 1d at a concentration of 1*10-4 mol/L inhibit Rho-kinase-2 by 26.72% and 18.11%, respectively.

Conclusion: The most active compounds, 7a and 1d, were identified as Rho kinase-2 inhibitors that exhibit ocular hypotensive effects in normotensive animals in vivo.

Graphical Abstract

Keywords:

glaucoma, benzimidazole, azepan, biphenyl, intraocular pressure, Rho-kinase (ROCK)

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Author Contribution

Olga N. Zhukovskaya, Southern Federal University, Rostov-on-Don, Russia

PhD in Chemical Sciences, Senior researcher at the Laboratory of Organic Synthesis, Institute of Physical and Organic Chemistry, Southern Federal University, Rostov-on-Don, Russia; e-mail: ol@yandex.ru; ORCID ID: http://orcid.org/0000-0003-2485-2139. The author proposed the design of the structures, synthesized the compounds, studied their structure, and wrote the chemical part of text.

Alena S. Taran, Volgograd State Medical University, Volgograd, Russia

PhD of Medical Sciences, Associate Professor, Associate Professor of the Department of Pharmacology and Bioinformatics, Volgograd State Medical University of the Ministry of Health of Russia, Volgograd, Russia; e-mail: alena-beretta-taran@mail.ru; ОRCID ID: https://orcid.org/0000-0001-8477-254X. The author designed the experimental study, participated in the experimental design, and wrote the pharmacologic part of the manuscript.

Lyudmila V. Naumenko, Volgograd State Medical University, Volgograd, Russia

Doctor Habil. of Medical Sciences, Associate Professor, Professor of the Department Pharmacology and Bioinformatics, Volgograd State Medical University of the Ministry of Health of Russia, Volgograd, Russia; e-mail: milanaumenko@mail.ru; ORCID ID: https://orcid.org/0000-0002-2119-4233. The author developed the design of the experimental study.

Alina M. Chebanko, Volgograd State Medical University, Volgograd, Russia

Assistant of the Department of Pharmacology and Bioinformatics, Volgograd State Medical University of the Ministry of Health of Russia, Volgograd, Russia; e-mail: chebanko@yandex.ru; ORCID ID: https://orcid.org/0009-0004-3140-5040. The author took part in conducting experimental work and analyzing the material.

Umida M. Ibragimovа, Volgograd State Medical University, Volgograd, Russia

Assistant of the Department of Pharmacology and Bioinformatics, Volgograd State Medical University of the Ministry of Health of Russia, Volgograd, Russia; e-mail: iumida@list.ru; ORCID ID: https://orcid.org/0000-0001-9141-4417. The author took part in conducting experimental work and analyzing the material.

Anatoly S. Morkovnik, Southern Federal University, Rostov-on-Don, Russia

Doctor Habil. of Chemical Sciences, Head of the Laboratory of Organic Synthesis, Institute of Physical and Organic Chemistry, Southern Federal University, Rostov-on-Don, Russia; e-mail: asmork@mail.ru; ORCID ID: https://orcid.org/0000-0002-9182-6101. The author was engaged in editing the chemical part of the manuscript.

Alexander A. Spasov, Volgograd State Medical University, Volgograd, Russia

Doctor Habil. of Medical Sciences, Professor, Member of the Russian Academy of Sciences, Head of the Department of Pharmacology and Bioinformatics, Volgograd State Medical University, Volgograd, Russia; e-mail: aspasov@mail.ru; ORCID ID: http://orcid.org/0000-0002-7185-4826. The author proposed the concept and design of the manuscript and approved of the final version of the manuscript.

 

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Published

26-06-2026

How to Cite

Zhukovskaya ON, Taran AS, Naumenko LV, Chebanko AM, Ibragimovа UM, Morkovnik AS, Spasov AA (2026) Synthesis and ophthalmic hypotensive effect of new potential benzimidazole-based Rho-kinase-2 inhibitors. Research Results in Pharmacology 12(2): 131–146. https://doi.org/10.18413/rrpharmacology.12.1121

Issue

Vol. 12 No. 2 (2026)

Section

Experimental Pharmacology

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Copyright (c) 2026 Zhukovskaya ON, Taran AS, Naumenko LV, Chebanko AM, Ibragimova UM, Morkovnik AS, Spasov AA

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