Pharmacological Reports最新文献

Background: Neuromuscular blocking agents (NMBAs) may trigger severe perioperative hypersensitivity reactions. A recently proposed mechanism involves off-target activity of NMBAs. They are thought to directly activate the Mas-related G protein-coupled receptor X2 (MRGPRX2), leading to mast cell degranulation and drug-induced hypersensitivity reactions. This study investigated which NMBAs exert this effect, whether in silico findings are consistent with cell-based experiments, and how the affinity of NMBAs for MRGPRX2 compares with that of fluoroquinolones, known MRGPRX2 agonists. We also sought to predict MRGPRX2 binding-site mutations that might enhance interactions with these drugs.

Methods: Molecular docking, molecular dynamics (MD) simulations, and molecular mechanics Poisson-Boltzmann surface area binding free-energy calculations were combined with a β-hexosaminidase release assay in MRGPRX2-expressing RBL-2H3 stable cell line. Computational alanine scanning was performed to predict the impact of receptor mutations.

Results: We demonstrated that atracurium-induced mast cell degranulation can be attributed to its immediate metabolite, laudanosine, which binds strongly to MRGPRX2. Pipecuronium, but not rocuronium, suxamethonium, or vecuronium, exhibited high affinity for MRGPRX2 in MD simulations. Complexes of ciprofloxacin, levofloxacin, and moxifloxacin with MRGPRX2 demonstrated excellent stability in MD simulations. RBL-MX2 responses to NMBAs and fluoroquinolones were highly consistent with our MD findings. Alanine substitutions reduced the affinity of ligands for MRGPRX2.

Conclusions: In contrast to fluoroquinolones, NMBAs displayed different affinity for MRGPRX2. Cellular responses in bench experiments closely reflected the MD predictions. Alanine scanning revealed that the MRGPRX2 binding pocket exhibits low susceptibility to single-site mutations that enhance receptor responsiveness.

Background: The synthesis of (3-phenylcarbamoyl-3,4-dihydro-2H-pyrrol-2-yl)phosphonates provided novel compounds with relevant affinities for imidazoline 2 (I2) receptors in human brain tissues. A selected compound, 12d, showed improved cognitive and analgesic properties at the preclinical level through the modulation of I2 receptors, but its potential innovative use as an antidepressant is still unknown.

Methods: Male and female adult Sprague-Dawley rats were treated with 3 ip injections of compound 12d (10 or 20 mg/kg), or vehicle (1 ml/kg DMSO), 24, 5, and 1 h prior to scoring its antidepressant-like efficacy under the stress of the forced-swim test. Hippocampal neuroplasticity markers (i.e., FADD, p-ERK/ERK, mBDNF) were evaluated 24 h post-treatment (and post-forced-swim test exposure).

Results: The novel results proved a sex-dependent efficacy of 12d, with dose-dependent antidepressant-like effects in adult male rats, and an inefficacious response for females. Moreover, compound 12d did not alter any of the hippocampal markers evaluated.

Conclusions: These results presented 12d as a novel therapeutic antidepressant candidate at the tested conditions, although only for male rats, thus requiring further studies to better understand the observed sex disparities as well as its molecular underpinnings. Moreover, compound 12d joins the list of other I2 receptor ligands with known antidepressant-like efficacy, validating and strengthening this receptor as a target in this field for future drug development.

Background: Systemic sclerosis (SSc) is an autoimmune disease characterized by vasculopathy, fibrosis, and dysregulation of the immune system. The therapeutic options available for SSc have limited efficacy, and there is a clear need for new pharmacological alternatives. Thiazolidinediones (TZD) are molecules that have therapeutic potential for SSc due to their pharmacological properties. In this study, we aimed to evaluate the immunomodulatory activity of a new TZD analogue (LPSF/CR-35) in peripheral blood mononuclear cells (PBMC) from SSc patients.

Materials and methods: PBMC response to LPSF/CR-35 (100µM) was examined from SSc patients (n = 19) and healthy control subjects (n = 8) following PBMC stimulation with anti-CD3/CD28. Cytokines (IFN-γ, TNF, IL-2, IL-4, IL-6, IL-10, and IL-17 A) and chemokines (CXCL8/IL-8, CCL5/RANTES, CXCL9/MIG, CCL2/MCP-1, and CXCL10/IP-10) levels were quantified in PBMC culture supernatants by cytometric bead array (CBA). In silico analyses were performed to evaluate the binding affinity of LPSF/CR-35 to peroxisome proliferator-activated receptor gamma (PPARγ).

Results: LPSF/CR-35 treatment significantly reduced the secretion of IL-17 A (p = 0.02), IL-10 (p = 0.001), IL-4 (p = 0.04), CCL2/MCP-1 (p = 0.003), and CXCL8/IL-8 (p = 0.03), while increasing TNF levels (p = 0.004) in PBMCs from SSc patients. Molecular docking predicted high binding affinity of both E (-9.987 kcal/mol) and Z (-9.992 kcal/mol) isomers of LPSF/CR-35 to PPARγ, supporting a possible PPARγ-dependent mechanism.

Conclusions: Our results indicate that LPSF/CR-35 modulates key cytokines and chemokines involved in SSc immunopathology, possibly through PPARγ activation. These findings support the potential of LPSF/CR-35 as an immunomodulatory agent in SSc. Further studies are needed to elucidate its effects on fibrosis and confirm its mechanism of action.

Background: The proper functioning of the nervous system determines the homeostasis of the entire body. There are many known approaches designed to positively stimulate the functions of the central nervous system by applying various plants and fungal extracts, but their course of action is poorly understood. Hericium erinaceus and Ganoderma lucidum are examples of fungi with medicinal properties and with a positive health-promoting effect. Therefore, the aim of our study was to evaluate the effect of H. erinaceus or G. lucidum M-CFS with their active metabolites alone and/or in co-treatment with CAY10464 [antagonist of aryl hydrocarbon receptor (AhR)] on the metabolic parameters, cell cycle, and selected protein expression.

Methods: The study was based on the use of the resazurin reduction assay, flow cytometry analyses, and Western blotting in the mouse hippocampal neuronal cell line (HT-22) in vitro.

Results: The obtained results proved no cytotoxicity of the tested metabolites towards the HT-22 cells in the concentration range of 2.5% - 10% of culture medium. The cells treated with the tested compounds were characterized by an increase in the protein expression of SQSTM/p62, PCNA, c-SRC, SOD1, AhR, Beclin 1, and ERK1/2. Moreover, a significant role of AhR in the mechanism of action of the tested metabolites was observed at the protein expression level.

Conclusion: The observed increase in the proliferation-related markers in the HT-22 cells proves the beneficial protective potential of these M-CFSs. Given the findings, we speculate their positive impact on the cognitive functions in the central nervous system.

Clinical trial registration date: Not applicable.

Clinical trial number: Not applicable.