Nadeem Irshad

Amide-linked chalcone derivatives were designed, synthesized and evaluated using in silico , in vitro and in vivo studies for their potential to relieve pain and inflammation.

Multiple sclerosis (MS) has a complex pathophysiology, comprising inflammation, demyelination, and alterations in neurotransmitter levels. Dysregulation of neurotransmitters, an imbalance between GABAergic and glutamatergic activity, enhanced oxidative stress, and an inflammatory response have all been involved in the disease. These mechanisms play a crucial role in regulating neuronal plasticity and activity. This study explored the role of neurotransmitters and inflammatory mediators in experimental autoimmune encephalomyelitis (EAE)-induced MS, as well as the effect of a 1,3,4-oxadiazole derivative, i.e., 2-{[5-(3-iodophenyl)-1,3,4-oxadiazol-2-yl] sulfanyl} ethan-1-ol (abbreviated as OX-3), on reducing neurotransmitter-mediated inflammatory responses. Its activity was assessed using clinical scoring, behavioral testing, histopathological changes, molecular testing through RT-PCR, ELISA, and assessment of neurotransmitters through HPLC. Molecular docking and simulation studies were performed to analyze protein interactions. The study findings revealed that OX-3 significantly reduced the severity of the disease by improving clinical symptoms of MS. It also ameliorated the cognitive decline in MS through a significant impact on neurotransmitters. It improved the GABA/Glutamate balance and reduced the pro-inflammatory cytokine levels. The comet assay revealed a reduction in DNA damage, and antioxidant levels improved, decreasing oxidative stress. The study concludes that OX-3 exerts its neuroprotective effect by attenuating the neurotransmitter-mediated neurodegeneration.

In the present study, a chemical library of novel lophine clubbed acylthioureas (8a–l) was designed and synthesized through a multistep sequence. 2-(4-Nitrophenyl)-4,5-diphenyl-1H-imidazole (3) was synthesized from benzil, 4-nitrobenzaldehyde, and ammonium acetate via multicomponent reaction. Then the nitro group of the synthesized compound was reduced using zinc and hydrochloric acid to furnish 4-(4,5-diphenyl-1H-imidazol-2-yl) aniline (4). On the other hand, the acid chloride (6a–l) of substituted benzoic acids was prepared by reacting acids (5a–l) with thionyl chloride, followed by their reaction with potassium thiocyanate to afford corresponding acyl isothiocyanates (7a–l), which were further reacted with 4-(4,5-diphenyl-1H-imidazol-2-yl)aniline (4) to produce acylthioureas (8a–l) in good yield. The synthesized series (8a–l) was characterized by using a combination of spectroscopic techniques, including FT-IR, ¹H NMR, and ¹³C NMR. Moreover, the synthesized acylthiourea derivatives were assayed for their in vitro and in silico xanthine oxidase and thymidine phosphorylase inhibitory activities. In the tested series, compounds 8j, 8d, and 8i showed robust dual inhibition profiles in low micromolar ranges. Among them, compound 8j revealed the most potent activity, with IC₅₀ values of 1.87 ± 0.26 µM against xanthine oxidase and 1.62 ± 0.27 µM against thymidine phosphorylase. Compound 8d sits in the next position, with an IC₅₀ value of 3.34 ± 0.37 µM (xanthine oxidase) and 3.31 ± 0.06 µM (thymidine phosphorylase), while compound 8i demonstrated considerable xanthine oxidase inhibition with an IC₅₀ of 2.06 ± 0.36 µM. These values have outperformed the standard drug allopurinol (IC₅₀ = 7.4 ± 0.07 µM), which was found to be less potent against xanthine oxidase. Considering the established involvement of these enzymes in oxidative stress, hyperuricemia, and inflammation-based disorders, the results endorse the idea of these derivatives being promising multifunctional therapeutic agents in the treatment of metabolic, inflammatory, and neoplastic diseases.

Gastric ulcer (GU) is primarily caused by Helicobacter pylori (H. pylori) infection, ethanol-induced oxidative stress, and excessive gastric acid secretion. This is a significant global health burden. Lifetime prevalence of peptic ulcer disease (including GU and duodenal ulcer) in the general population has been estimated to be about 5–10%, and incidence 0.1–0.3% per year. Kahweol (a coffee-derived diterpene) exhibits broad pharmacological effects. Although Kahweol has strong antioxidant, anti-inflammatory and cytoprotective effects, its gastroprotective potential against ethanol-induced GU has remained unexplored. This is the first study demonstrating the multifaceted gastroprotective effects of Kahweol using an integrated computational, in vitro, and in vivo approaches, including direct comparison with omeprazole and evidence of dual acid-suppressive and anti-H. pylori activity. Molecular docking and 100 ns molecular dynamics simulations demonstrated stable Kahweol binding to H⁺/K⁺-ATPase, nuclear factor-kappa B, NOD-like receptor protein 3, and cyclooxygenase-2. Kahweol exhibits inhibition of inflammatory pathways and acid secretion, having the highest affinity for H⁺/K⁺-ATPase (-7.92 kcal/mol). In vitro assays revealed that Kahweol has dose-dependent antibacterial activity against H. pylori clinical isolates. In vivo, Kahweol significantly decreased the ethanol-induced ulcer index in a dose-dependent manner. Kahweol pretreatment inhibited GU up to 80% with 10 mg/kg dose, which was comparable to that of omeprazole (85%). Histopathological analysis in Kahweol-treated groups confirmed preserved mucosal architecture, reduced vacuolation, and restored epithelial boundaries, which were comparable to those of omeprazole. Biochemical assays of Kahweol-treated groups showed enhanced antioxidant defenses, with increased levels of reduced glutathione, glutathione S-transferase, and catalase, along with decreased levels of nitric oxide and malondialdehyde. Results from the enzyme-linked immunosorbent assay of the Kahweol-treated group showed that the pro-inflammatory cytokines interleukin-1 beta and tumor necrosis factor-alpha were suppressed, confirming anti-inflammatory effects. Quantitative real-time polymerase chain reaction verified that the expression of H⁺/K⁺-ATPase was downregulated by Kahweol, indicating the acid-inhibition effect of Kahweol. These findings suggest Kahweol is the potential natural agent for the prevention and treatment of ethanol-induced GU, highlighting the comprehensive gastroprotective effects of Kahweol through its antioxidant, anti-inflammatory, anti-secretory, and anti-H. pylori activity.

1,3,4-Oxadiazole derivatives have attracted substantial attention as promising therapeutic agents for neurodegenerative disorders due to their anti-inflammatory and neuroprotective properties. This study specifically examined a 1,3,4-oxadiazole derivative, i.e.,-{[5-(3-bromophenyl)-1, 3, 4-oxadiazol-2-yl] sulfanyl} ethan-1-ol (abbreviated as OX-1), in the context of multiple sclerosis (MS). Currently, approximately 2.8 million individuals worldwide are living with MS. The study demonstrates the therapeutic potential of OX-1 using the experimental autoimmune encephalomyelitis (EAE) model of MS. Neurobehavioral assessments indicate significant improvements in clinical scoring, motor deficits, muscular strength, and locomotor activity in EAE subjects. Imaging test reveal notable improvements in spinal deformity, further supporting the efficacy of the compound. Comprehensive analyses, including antioxidant assays, RT-PCR, and comet assays, confirm that OX-1 effectively reduces oxidative stress, accompanied by a significant decrease in cytokine expression. Histological examinations reveal critical pathological changes in the hippocampus, cortex, eyes, spinal cord, and optic nerve. The data demonstrate that this compound exhibits neuroprotective effects by activating the Nrf2/HO-1 pathway, thereby reducing oxidative stress and enhancing the antioxidant defense system. Additionally, it suppresses the TLR4/NF-κB pathway, significantly lowering pro-inflammatory cytokine production and immune cell infiltration. Furthermore, molecular docking and simulation studies demonstrate the binding interactions and potential modulatory effects of OX-1 on HO-1 and NF-κB. These results emphasize the therapeutic promise of OX-1 in effectively alleviating the clinical signs and symptoms associated with EAE-induced MS. Graphical Abstract The graphical abstract of EAE model of MS.

Chronic inflammatory disorders are often accompanied by persistent pain, yet current therapies remain limited by modest efficacy and adverse effects. To address this challenge, a series of eleven chalcone-based derivatives was synthesized via Claisen–Schmidt condensation and structurally characterized by FTIR, ¹H-NMR, ¹³C-NMR, and EIMS analyses. Cytotoxicity assessment on 3T3 fibroblasts using the MTT assay revealed no toxicity compared to doxorubicin, while brine shrimp lethality and hemolytic assays further confirmed their safety profile. In vitro COX-1/2 assays demonstrated preferential inhibition of COX-2, with AS1-6 showing a superior selectivity index relative to nimesulide. In vivo screening corroborated these results: AS1-2, AS1-6, and AS1-8 produced significant anti-inflammatory effects in carrageenan-induced paw edema, while AS1-2, AS1-6, and AS1-10 suppressed nociceptive behaviours in the formalin test, indicating both peripheral and central antinociceptive activity. Ex vivo ELISA further verified down-regulation of COX-2-associated inflammatory mediators compared to negative group. Molecular docking revealed strong and selective interactions of AS1-2 and AS1-6 within the COX-2 active site, while sparing COX-1. Molecular dynamics simulations confirmed the stability of AS1-6 in the active pocket throughout a 100 ns trajectory, consistent with the pharmacological outcomes. Collectively, the concordance of in vitro COX-1/2 inhibition, in vivo efficacy, and in silico analyses underscores the potential of these chalcone analogues as selective COX-2 inhibitors. These findings highlight the value of structure-guided design in developing dual-function chalcones with anti-inflammatory and antinociceptive activity, warranting further optimization and preclinical evaluation.