PMC

(A–L) Lateral views of 6 day post-fertilization larvae following fixation and staining with oil red o to mark neutral lipid. (A) Unfed wild type larva. (B) Wild type larva fed 1% egg yolk. Arrows point to lipid in vasculature. (C–I) Compound treated larvae (25 uM) fed 1% egg yolk. There is reduced intestinal lipid in the compound treated larvae, with no evidence of lipid in the vasculature. Hepatic lipid accumulation is not detected. Lipid is present in the intestinal lumen of larvae treated with compound 10 and ezetimibe (50 uM; blue asterisk).

(Upper panel) Analysis of 2D TOCSY spectra collected at 800 MHz of the aromatic region of compound BAS00127538 alone (black) overlaid with spectra of compound bound to Lipid II (red) (Lower panel). 2D natural abunance ¹³C HSQC spectrum illustrating the interaction between Lipid II and the compound BAS00127538. BAS00127538 alone (black) is overlaid with a spectrum of compound bound to Lipid II (red). Spectra were collected on a Bruker 800 MHz Avance NMR spectrometer at 25 degrees. Chemical shift changes for Lipid II upon BAS00127538 compound binding suggest that the interaction is occuring at or near the MurNAc moeity of Lipid II.

Schemes for synthesis of a lipid mimetic of GM3, lyso-GM3, and a lyso-GM3 dimer
Scheme 1. Synthesis of lipid mimetic of lyso-GM3. Reagents and conditions: (a) Ac₂O, pyridine, DMF, 50 °C, 24 h, 83.7%; (b) 2-azido-1-dodecanol, BF₃·(C₂H₅)₂O, molecular sieves, 1,2-dichloroethane, 40 °C, 24 h, 31.7%; (c) PPh₃, H₂O, DMF, 50 °C, 20h, 91.6%.
Scheme 2. Synthesis of lipid mimetic of GM3 (LM-GM3). Reagents and conditions: (a) compound 3 was reacted with stearic acid in EDC, DMAP, anhyd. DMF, at room temperature (r.t) for 17 h; the yield of compound 4, 67.9%; (b) deacetylated in NaOMe, MeOH, r.t., 16 h, the yield of compound 5 (i.e., 2-N-stearylaminododecyl-O-(5-acetamido-α-3,5-dideoxy-D-glycero-D-galacto-2-nonulopyranosylonic acid)-(2→3)-O-β-D-galactopyranosyl-(1→4)-β-D-glucopyranoside), 55.8%.
Scheme 3. Assembly of a lipid mimetic of lyso-GM3 with a glutamic acid dimmer (synthesis of a lipid mimetic of lyso-GM3 dimer; LM-dimer). Reagents and conditions: (a) compound 3 was reacted with Fmoc-(Glu)₂-NH₂ in PyBOP, DIEA, DMAP, anhyd. DMF, r.t, 16 h, the yield of compound 6, 92.6%; (b) NaOMe, MeOH, r.t., 14 h, the yield of compound 7, 66.8%. Details of conditions: see .
Abbreviations: DIEA, N,N-diisopropylethylamine; DMAP, 4-(N,N-dimethylamino)pyridine; DMF, dimethylformamide; EDC, 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride.

Effect of compound 1a on lipid adipogenesis. A: Effect of compound 1a on lipid accumulation in 3T3-L1 adipocytes. At two days after confluence, 3T3-L1 cells were differentiated by MDI. At day 8 compound 1a was given to the cells at series of concentrations. Ten days after differentiation, we measured triglyceride content of 3T3-L1 cells treated with compound 1a by Oil-Red-O staining. The inhibition ratio was calculated by formula 1. B: Effect of vehicle, compound 1a, Inositol Nicotinate on the lipid accumulation by Oil-Red-O staining. Images of the stained lipid droplets were collected on a fluorescence microscope. Lipid accumulation was significantly decreased treated by compound 1a. All values are mean ± SD (n = 3).