Journal: Pharmacological Reports

Article Title: In silico assessment of neuromuscular blocking agents and fluoroquinolones as ligands of the Mas-related G protein-coupled receptor X2

doi: 10.1007/s43440-025-00813-7

Figure Lengend Snippet: ( A ) Ciprofloxacin docking pose as predicted with GNINA. ( B ) Schematic breakdown of protein-ligand interactions between ciprofloxacin and MRGPRX2 binding pocket residues. Colours of the dashed lines denote; hydrophobic interactions (green), cationic interactions (red), hydrogen bonds (blue), aromatic interactions (violet). ( C ) Superimposed docking poses of studied fluoroquinolones (ciprofloxacin – yellow, levofloxacin - green, dextrofloxacin - blue, moxifloxacin - pink). ( D ) RMSD (root mean square deviation) plot of the fluoroquinolone ligands during 100 ns of molecular dynamics simulation in complex with MRGPRX2. The presented values of RMSD are calculated as the mean of three separate MD runs

Article Snippet: The following day, the monolayers were washed twice with extracellular buffer (EXB) [ ] and stimulated for 60 min at 37 °C in EXB containing either the reference MRGPRX2 agonist (R)-ZINC-3573 (Tocris, Cat. No. 6351), at concentrations of 1, 5 or 10 μM, the fluoroquinolones: ciprofloxacin (100, 250–500 μg/ml; Ciprofloxacin Kabi , Fresenius Kabi), moxifloxacin (50, 100–250 μg/ml; Sigma-Aldrich, Cat. No. SML1581) and levofloxacin (100, 500–1000 μg/ml; Levoxa , Actavis Group), or the neuromuscular blocking agents: atracurium (200, 500–1000 μg/ml; Tracrium , Aspen Pharma), pipecuronium (100, 250, 500 μg/ml; Sigma-Aldrich, Cat. No. SML1636), rocuronium (500, 1000–2000 μg/ml; Rocuronium Kabi , Fresenius Kabi), vecuronium (500, 1000–2000 μg/ml; Sigma-Aldrich, Cat. No. 76904) and suxamethonium (250, 500–1000 μg/ml; Chlorsuccillin , Bausch Health).

Techniques: Binding Assay

Journal: Pharmacological Reports

Article Title: In silico assessment of neuromuscular blocking agents and fluoroquinolones as ligands of the Mas-related G protein-coupled receptor X2

doi: 10.1007/s43440-025-00813-7

Figure Lengend Snippet: β-hexosaminidase release from RBL-MX2 cells expressed as relative to that from parental RBL-2H3 cells ( A ) for negative control (extracellular buffer), CTRL(-); (R)-ZINC-3573, ZINC; the investigated fluoroquinolones: ciprofloxacin, CIP; moxifloxacin, MOX; levofloxacin, LEV; and ( B ) neuromuscular blocking agents: atracurium, ATR; pipecuronium, PIP; rocuronium, ROC; vecuronium, VEC and suxamethonium, SUX in consecutive concentrations. Diagonal stripes indicate the concentration eliciting the strongest response. Data derived from of at least three independent experiments. The medians with interquartile ranges are shown. Statistical analyses were performed using a two-way Mann-Whitney test. Statistical significance is indicated as **** p < 0.0001; *** p ≤ 0.001. RBL-2H3, rat basophilic leukaemia cells; RBL-MX2, RBL-2H3 cells expressing Mas-related G protein-coupled receptor X2

Article Snippet: The following day, the monolayers were washed twice with extracellular buffer (EXB) [ ] and stimulated for 60 min at 37 °C in EXB containing either the reference MRGPRX2 agonist (R)-ZINC-3573 (Tocris, Cat. No. 6351), at concentrations of 1, 5 or 10 μM, the fluoroquinolones: ciprofloxacin (100, 250–500 μg/ml; Ciprofloxacin Kabi , Fresenius Kabi), moxifloxacin (50, 100–250 μg/ml; Sigma-Aldrich, Cat. No. SML1581) and levofloxacin (100, 500–1000 μg/ml; Levoxa , Actavis Group), or the neuromuscular blocking agents: atracurium (200, 500–1000 μg/ml; Tracrium , Aspen Pharma), pipecuronium (100, 250, 500 μg/ml; Sigma-Aldrich, Cat. No. SML1636), rocuronium (500, 1000–2000 μg/ml; Rocuronium Kabi , Fresenius Kabi), vecuronium (500, 1000–2000 μg/ml; Sigma-Aldrich, Cat. No. 76904) and suxamethonium (250, 500–1000 μg/ml; Chlorsuccillin , Bausch Health).

Techniques: Negative Control, Blocking Assay, Concentration Assay, Derivative Assay, MANN-WHITNEY, Expressing

Journal: Pharmacological Reports

Article Title: In silico assessment of neuromuscular blocking agents and fluoroquinolones as ligands of the Mas-related G protein-coupled receptor X2

doi: 10.1007/s43440-025-00813-7

Figure Lengend Snippet: Results of computational alanine scan for (R)- and (S)-laudanosine, pipecuronium, ciprofloxacin and the selective MRGPRX2 agonist, (R)-ZINC-3573. The bars illustrate the change in binding free energy ΔΔG bind [kcal/mol] upon substitution of binding pocket residues with alanine

Article Snippet: The following day, the monolayers were washed twice with extracellular buffer (EXB) [ ] and stimulated for 60 min at 37 °C in EXB containing either the reference MRGPRX2 agonist (R)-ZINC-3573 (Tocris, Cat. No. 6351), at concentrations of 1, 5 or 10 μM, the fluoroquinolones: ciprofloxacin (100, 250–500 μg/ml; Ciprofloxacin Kabi , Fresenius Kabi), moxifloxacin (50, 100–250 μg/ml; Sigma-Aldrich, Cat. No. SML1581) and levofloxacin (100, 500–1000 μg/ml; Levoxa , Actavis Group), or the neuromuscular blocking agents: atracurium (200, 500–1000 μg/ml; Tracrium , Aspen Pharma), pipecuronium (100, 250, 500 μg/ml; Sigma-Aldrich, Cat. No. SML1636), rocuronium (500, 1000–2000 μg/ml; Rocuronium Kabi , Fresenius Kabi), vecuronium (500, 1000–2000 μg/ml; Sigma-Aldrich, Cat. No. 76904) and suxamethonium (250, 500–1000 μg/ml; Chlorsuccillin , Bausch Health).

Techniques: Binding Assay

Journal: Pharmacological Reports

Article Title: In silico assessment of neuromuscular blocking agents and fluoroquinolones as ligands of the Mas-related G protein-coupled receptor X2

doi: 10.1007/s43440-025-00813-7

Figure Lengend Snippet: The positioning of ciprofloxacin, (R)-laudanosine and (S)-laudanosine with respect to the hydrophobic groove formed by L247 and W248 residues

Article Snippet: The following day, the monolayers were washed twice with extracellular buffer (EXB) [ ] and stimulated for 60 min at 37 °C in EXB containing either the reference MRGPRX2 agonist (R)-ZINC-3573 (Tocris, Cat. No. 6351), at concentrations of 1, 5 or 10 μM, the fluoroquinolones: ciprofloxacin (100, 250–500 μg/ml; Ciprofloxacin Kabi , Fresenius Kabi), moxifloxacin (50, 100–250 μg/ml; Sigma-Aldrich, Cat. No. SML1581) and levofloxacin (100, 500–1000 μg/ml; Levoxa , Actavis Group), or the neuromuscular blocking agents: atracurium (200, 500–1000 μg/ml; Tracrium , Aspen Pharma), pipecuronium (100, 250, 500 μg/ml; Sigma-Aldrich, Cat. No. SML1636), rocuronium (500, 1000–2000 μg/ml; Rocuronium Kabi , Fresenius Kabi), vecuronium (500, 1000–2000 μg/ml; Sigma-Aldrich, Cat. No. 76904) and suxamethonium (250, 500–1000 μg/ml; Chlorsuccillin , Bausch Health).

Techniques:

Journal: Pharmacological Reports

Article Title: In silico assessment of neuromuscular blocking agents and fluoroquinolones as ligands of the Mas-related G protein-coupled receptor X2

doi: 10.1007/s43440-025-00813-7

Figure Lengend Snippet: Molecular structures of the studied neuromuscular junction blockers – rocuronium, vecuronium, pipecuronium, atracurium (with its major metabolite, laudanosine) and suxamethonium (succinylcholine)

Article Snippet: The following day, the monolayers were washed twice with extracellular buffer (EXB) [ ] and stimulated for 60 min at 37 °C in EXB containing either the reference MRGPRX2 agonist (R)-ZINC-3573 (Tocris, Cat. No. 6351), at concentrations of 1, 5 or 10 μM, the fluoroquinolones: ciprofloxacin (100, 250–500 μg/ml; Ciprofloxacin Kabi , Fresenius Kabi), moxifloxacin (50, 100–250 μg/ml; Sigma-Aldrich, Cat. No. SML1581) and levofloxacin (100, 500–1000 μg/ml; Levoxa , Actavis Group), or the neuromuscular blocking agents: atracurium (200, 500–1000 μg/ml; Tracrium , Aspen Pharma), pipecuronium (100, 250, 500 μg/ml; Sigma-Aldrich, Cat. No. SML1636), rocuronium (500, 1000–2000 μg/ml; Rocuronium Kabi , Fresenius Kabi), vecuronium (500, 1000–2000 μg/ml; Sigma-Aldrich, Cat. No. 76904) and suxamethonium (250, 500–1000 μg/ml; Chlorsuccillin , Bausch Health).

Techniques:

Journal: Pharmacological Reports

Article Title: In silico assessment of neuromuscular blocking agents and fluoroquinolones as ligands of the Mas-related G protein-coupled receptor X2

doi: 10.1007/s43440-025-00813-7

Figure Lengend Snippet: RMSD (root mean square deviation) of the studied NMBAs atom coordinates over 100 ns of MD production run, calculated with respect to the starting (docked) pose. Pipecuronium, (R)- and (S)-laudanosine exhibit excellent stability in complex with MRGPRX2, while rocuronium, vecuronium and succinylcholine dissociate readily

Article Snippet: The following day, the monolayers were washed twice with extracellular buffer (EXB) [ ] and stimulated for 60 min at 37 °C in EXB containing either the reference MRGPRX2 agonist (R)-ZINC-3573 (Tocris, Cat. No. 6351), at concentrations of 1, 5 or 10 μM, the fluoroquinolones: ciprofloxacin (100, 250–500 μg/ml; Ciprofloxacin Kabi , Fresenius Kabi), moxifloxacin (50, 100–250 μg/ml; Sigma-Aldrich, Cat. No. SML1581) and levofloxacin (100, 500–1000 μg/ml; Levoxa , Actavis Group), or the neuromuscular blocking agents: atracurium (200, 500–1000 μg/ml; Tracrium , Aspen Pharma), pipecuronium (100, 250, 500 μg/ml; Sigma-Aldrich, Cat. No. SML1636), rocuronium (500, 1000–2000 μg/ml; Rocuronium Kabi , Fresenius Kabi), vecuronium (500, 1000–2000 μg/ml; Sigma-Aldrich, Cat. No. 76904) and suxamethonium (250, 500–1000 μg/ml; Chlorsuccillin , Bausch Health).

Techniques:

Journal: Pharmacological Reports

Article Title: In silico assessment of neuromuscular blocking agents and fluoroquinolones as ligands of the Mas-related G protein-coupled receptor X2

doi: 10.1007/s43440-025-00813-7

Figure Lengend Snippet: β-hexosaminidase release from RBL-MX2 cells expressed as relative to that from parental RBL-2H3 cells ( A ) for negative control (extracellular buffer), CTRL(-); (R)-ZINC-3573, ZINC; the investigated fluoroquinolones: ciprofloxacin, CIP; moxifloxacin, MOX; levofloxacin, LEV; and ( B ) neuromuscular blocking agents: atracurium, ATR; pipecuronium, PIP; rocuronium, ROC; vecuronium, VEC and suxamethonium, SUX in consecutive concentrations. Diagonal stripes indicate the concentration eliciting the strongest response. Data derived from of at least three independent experiments. The medians with interquartile ranges are shown. Statistical analyses were performed using a two-way Mann-Whitney test. Statistical significance is indicated as **** p < 0.0001; *** p ≤ 0.001. RBL-2H3, rat basophilic leukaemia cells; RBL-MX2, RBL-2H3 cells expressing Mas-related G protein-coupled receptor X2

Article Snippet: The following day, the monolayers were washed twice with extracellular buffer (EXB) [ ] and stimulated for 60 min at 37 °C in EXB containing either the reference MRGPRX2 agonist (R)-ZINC-3573 (Tocris, Cat. No. 6351), at concentrations of 1, 5 or 10 μM, the fluoroquinolones: ciprofloxacin (100, 250–500 μg/ml; Ciprofloxacin Kabi , Fresenius Kabi), moxifloxacin (50, 100–250 μg/ml; Sigma-Aldrich, Cat. No. SML1581) and levofloxacin (100, 500–1000 μg/ml; Levoxa , Actavis Group), or the neuromuscular blocking agents: atracurium (200, 500–1000 μg/ml; Tracrium , Aspen Pharma), pipecuronium (100, 250, 500 μg/ml; Sigma-Aldrich, Cat. No. SML1636), rocuronium (500, 1000–2000 μg/ml; Rocuronium Kabi , Fresenius Kabi), vecuronium (500, 1000–2000 μg/ml; Sigma-Aldrich, Cat. No. 76904) and suxamethonium (250, 500–1000 μg/ml; Chlorsuccillin , Bausch Health).

Techniques: Negative Control, Blocking Assay, Concentration Assay, Derivative Assay, MANN-WHITNEY, Expressing