Journal: Scientific Reports

Article Title: Novel carboxylate-based glycolipids: TLR4 antagonism, MD-2 binding and self-assembly properties

doi: 10.1038/s41598-018-37421-w

Figure Lengend Snippet: ( A ) Monosaccharide FP13-17 are lipid A mimetics. ( B ) FP13 contains two oleic acid chains (C18, cis-9), FP14 two linoleic acid chains (C18, cis, cis-9,12), FP15 myristic acid at C3 (C14) and oleic acid at C2, FP16 myristic acid at C3 and linoleic acid at C2 and FP17 two myristic acid chains.

Article Snippet: Compounds FP13- FP17 (Fig. ) have been rationally designed as lipid A mimetics where the phosphates have been replaced by succinate moieties, while the distance between the two carboxylate groups has been kept similar to that of the two phosphates groups in lipid A (distance of 11 to 15 Å for E . coli Lipid A and 8 to 15 Å for FP13-17) – , – .

Techniques:

Journal: Scientific Reports

Article Title: Novel carboxylate-based glycolipids: TLR4 antagonism, MD-2 binding and self-assembly properties

doi: 10.1038/s41598-018-37421-w

Figure Lengend Snippet: Best AutoDock Vina docked poses of FP13-17 within the TLR4/MD-2 complex. TLR4 and MD-2 are respectively represented in black and grey cartoon. Ligands FP13 to FP17 are in yellow, cyan, pink, blue and green sticks, respectively (left: general view; right: detailed view showing selected interacting residues, only FP13 in yellow and FP16 in blue are shown).

Article Snippet: Compounds FP13- FP17 (Fig. ) have been rationally designed as lipid A mimetics where the phosphates have been replaced by succinate moieties, while the distance between the two carboxylate groups has been kept similar to that of the two phosphates groups in lipid A (distance of 11 to 15 Å for E . coli Lipid A and 8 to 15 Å for FP13-17) – , – .

Techniques:

Journal: Scientific Reports

Article Title: Novel carboxylate-based glycolipids: TLR4 antagonism, MD-2 binding and self-assembly properties

doi: 10.1038/s41598-018-37421-w

Figure Lengend Snippet: Binding studies on purified hMD-2 receptor. ( A ) FP13-17 prevent anti-human hMD-2 monoclonal antibody binding in a dose-dependent manner; ( B ) FP13-17 compete with biotin-LPS for hMD-2 binding; ( C ) FP13-17 dose-dependently inhibit the binding of bis-ANS to hMD-2;( D )-( H ). SPR analysis show direct interaction between FP13-17 and hMD-2; K D values are reported.

Article Snippet: Compounds FP13- FP17 (Fig. ) have been rationally designed as lipid A mimetics where the phosphates have been replaced by succinate moieties, while the distance between the two carboxylate groups has been kept similar to that of the two phosphates groups in lipid A (distance of 11 to 15 Å for E . coli Lipid A and 8 to 15 Å for FP13-17) – , – .

Techniques: Binding Assay, Purification

Journal: Scientific Reports

Article Title: Novel carboxylate-based glycolipids: TLR4 antagonism, MD-2 binding and self-assembly properties

doi: 10.1038/s41598-018-37421-w

Figure Lengend Snippet: Dose-dependent inhibition of LPS-triggered TLR4 pathway activation in HEK293-Blue hTLR4 cells. Cells were pre-incubated with increasing concentrations (0.1 to 10 µM) of compounds FP13-17 in serum-free DMEM medium and stimulated with LPS (100 ng/mL) after 15 min. SEAP levels in cell culture media were quantified after 16 hours as indicator of TLR4 activation. Data were normalized to stimulation with LPS alone and fitted to a sigmoidal 4 parameter logistic equation to obtain dose-effect curves (left panel). IC 50 values for FP13-17 are reported in the right panel. points represent the mean of percentage ± SEM of at least 3 independent experiments.

Article Snippet: Compounds FP13- FP17 (Fig. ) have been rationally designed as lipid A mimetics where the phosphates have been replaced by succinate moieties, while the distance between the two carboxylate groups has been kept similar to that of the two phosphates groups in lipid A (distance of 11 to 15 Å for E . coli Lipid A and 8 to 15 Å for FP13-17) – , – .

Techniques: Inhibition, Activation Assay, Incubation, Cell Culture

Journal: Scientific Reports

Article Title: Novel carboxylate-based glycolipids: TLR4 antagonism, MD-2 binding and self-assembly properties

doi: 10.1038/s41598-018-37421-w

Figure Lengend Snippet: Inhibition of LPS-induced TLR4 signaling in RAW 264.7-Blue cells. Cells were pre-incubated with the indicated concentrations of monosaccharides FP13-17 and stimulated with 1, 10 or 100 ng/mL E . coli O111:B4 LPS 30 min. later in serum-free DMEM medium. SEAP reporter levels in the cell supernatant were determined by QuantiBlue Assay 24 h later. Error bars represent the standard deviation of the mean of technical triplicates. The graph is representative of at least three independent experiments.

Article Snippet: Compounds FP13- FP17 (Fig. ) have been rationally designed as lipid A mimetics where the phosphates have been replaced by succinate moieties, while the distance between the two carboxylate groups has been kept similar to that of the two phosphates groups in lipid A (distance of 11 to 15 Å for E . coli Lipid A and 8 to 15 Å for FP13-17) – , – .

Techniques: Inhibition, Incubation, Standard Deviation

Journal: Scientific Reports

Article Title: Novel carboxylate-based glycolipids: TLR4 antagonism, MD-2 binding and self-assembly properties

doi: 10.1038/s41598-018-37421-w

Figure Lengend Snippet: Presence of serum or BSA neutralizes the antagonistic activity of monosaccharide FP13 on LPS-induced TLR4 signaling in RAW 264.7-Blue cells. A-C. Cells were pre-incubated with the indicated concentrations of monosaccharide FP13 in DMEM supplemented with 10% FBS ( A ), 4 mg/mL BSA ( B ) or 20 mg/ml BSA ( C ). 30 min later cells were stimulated with 1, 10 or 100 ng/mL E . coli O111:B4 LPS. SEAP reporter levels in the cell supernatant were determined by QuantiBlue Assay 24 h later. Error bars represent the standard deviation of the mean of biological triplicates. The graph is representative of at least three independent experiments.

Article Snippet: Compounds FP13- FP17 (Fig. ) have been rationally designed as lipid A mimetics where the phosphates have been replaced by succinate moieties, while the distance between the two carboxylate groups has been kept similar to that of the two phosphates groups in lipid A (distance of 11 to 15 Å for E . coli Lipid A and 8 to 15 Å for FP13-17) – , – .

Techniques: Activity Assay, Incubation, Standard Deviation