Mechanistic evaluation of a novel cyclohexenone derivative’s functionality against nociception and inflammation: An in-vitro, in-vivo and in-silico approach

• Published in: European journal of pharmacology 2021-07, Vol.902, p.174091, Article 174091
• Main Authors: Khan, Jawad, Ali, Gowhar, Rashid, Umer, Khan, Rasool, Jan, Muhammad Saeed, Ullah, Rahim, Ahmad, Sajjad, Abbasi, Sumra Wajid, Khan Khalil, Atif Ali, Sewell, RobertD.E.
• Format: Article
• Language: English
• Subjects: 5-Lipoxygenase; Analgesics - pharmacology; Analgesics - therapeutic use; Animals; Anti-inflammatory; Anti-Inflammatory Agents - pharmacology; Anti-Inflammatory Agents - therapeutic use; Anti-nociceptive; Arachidonate 5-Lipoxygenase - metabolism; Behavior, Animal - drug effects; Computer Simulation; Cyclohexanones - chemistry; Cyclohexanones - pharmacology; Cyclohexanones - therapeutic use; Cyclohexanones - toxicity; Cyclohexenes - chemistry; Cyclohexenes - pharmacology; Cyclohexenes - therapeutic use; Cyclohexenes - toxicity; Cyclohexenone; Cyclooxygenase 2 - chemistry; Cyclooxygenase 2 - genetics; Cyclooxygenase 2 - metabolism; Cyclooxygenase 2 Inhibitors - pharmacology; Cyclooxygenase 2 Inhibitors - therapeutic use; Cyclooxygenase 2 Inhibitors - toxicity; Cyclooxygenase-2; Cytokines - genetics; Cytokines - metabolism; Edema - chemically induced; Edema - drug therapy; Female; GABAA/Opioid receptors; Inflammation - chemically induced; Inflammation - drug therapy; Lipoxygenase Inhibitors - pharmacology; Lipoxygenase Inhibitors - therapeutic use; Lipoxygenase Inhibitors - toxicity; Male; Mice; Mice, Inbred BALB C; Nociceptive Pain - chemically induced; Nociceptive Pain - drug therapy; Receptors, GABA - chemistry; Receptors, GABA - drug effects; Receptors, Opioid - chemistry; Receptors, Opioid - drug effects
• ISSN: 0014-2999; 1879-0712
• DOI: 10.1016/j.ejphar.2021.174091

The synthesis of a novel cyclohexanone derivative (CHD; Ethyl 6-(4-metohxyphenyl)-2-oxo-4-phenylcyclohexe-3-enecarboxylate) was described and the subsequent aim was to perform an in vitro, in vivo and in silico pharmacological evaluation as a putative anti-nociceptive and anti-inflammatory agent in mice. Initial in vitro studies revealed that CHD inhibited both cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX) enzymes and it also reduced mRNA expression of COX-2 and the pro-inflammatory cytokines TNF-α and IL-1β. It was then shown that CHD dose dependently inhibited chemically induced tonic nociception in the abdominal constriction assay and also phasic thermal nociception (i.e. anti-nociception) in the hot plate and tail immersion tests in comparison with aspirin and tramadol respectively. The thermal test outcomes indicated a possible moderate centrally mediated anti-nociception which, in the case of the hot plate test, was pentylenetetrazole (PTZ) and naloxone reversible, implicating GABAergic and opioidergic mechanisms. CHD was also effective against both the neurogenic and inflammatory mediator phases induced in the formalin test and it also disclosed anti-inflammatory activity against the phlogistic agents, carrageenan, serotonin, histamine and xylene compared with standard drugs in edema volume tests. In silico studies indicated that CHD possessed preferential affinity for GABAA, opioid and COX-2 target sites and this was supported by molecular dynamic simulations where computation of free energy of binding also favored the formation of stable complexes with these sites. These findings suggest that CHD has prospective anti-nociceptive and anti-inflammatory properties, probably mediated through GABAergic and opioidergic interactions supplemented by COX-2 and 5-LOX enzyme inhibition in addition to reducing pro-inflammatory cytokine expression. CHD may therefore possess potentially beneficial therapeutic effectiveness in the management of inflammation and pain. [Display omitted] •A synthesized anti-inflammatory cyclohexanone (CHD) was tested in vivo and in vitro.•CHD inhibited COX-2 and 5-LOX enzymes plus COX-2, TNF-α and IL-1β mRNA expression.•CHD also produced GABAA and opioid mediated inhibitory activity in nociceptive tests.•In silico CHD had preferential affinity for GABAA, opioid and COX-2 target sites.•CHD may possess therapeutic effectiveness in the management of inflammation and pain.