By Vincenzo Di Marzo, PhD, Istituto di Chimica Biomolecolare Pozzuoli, NA, Italy.

1. Role of the Dysregulated Endocannabinoid System in Determining Cardiometabolic Risk Vincenzo Di Marzo, PhD Istituto di Chimica Biomolecolare Pozzuoli (NA), Italy

2. Endocannabinoids (Endogenous Agonists of Cannabinoid Receptors) 1) are produced “ on demand” 2) activate cannabinoid receptors locally 3) are immediately metabolized Phospholipid-derived precursors Endocannabinoids Degradation products NAPE-PLD DAG Lipase O 2-arachidonoylglycerol O Anandamide N H OH H 2 N OH O OH Fatty Acid Amide Hydrolase MAG Lipase NH O O Phospholipid Remodelling O OH HO OH CH OH O OH CH OH O O CH R 3 P O O O O R 2 R 1 O DAG Lipase : diacylglycerol lipase MAG Lipase : monoacylglycerol lipase NAPE-PLD : N-acylphosphatidyethanolamine-hydrolyzing phospholipase D

3. The CB1 Receptor Stimulates Adipocyte Differentiation and Lipogenesis Adapted from Matias I et al. J Clin Endocrinol Metab 2006; 91: 3171-80 Reproduced with permission *** p<0.005 vs. vehicle ## p<0.01 vs. HU210 HU210: cannabinoid receptor agonist SR1: rimonabant Forskolin 1  M HU210 20 nM HU210 200 nM SR1 20nM SR1 200nM HU210 SR1 20nM HU210 SR1 200nM % forskolin-induced cAMP inhibition

4. Overactive Endocannabinoid System in Adipose Tissue and Liver of Mice with Diet-Induced Obesity Adapted from Osei-Hyiaman D et al. J Clin Invest 2005; 115: 1298-1305 1.6 1.2 0.8 0.4 0.0 AEA: anandamide 2-AG: 2-arachidonoylglycerol Adapted from Matias I et al. J Clin Endocrinol Metab 2006; 91: 3171-80 Reproduced with permission Anandamide (pmol/g) 2-AG (nmol/g)

5. The Overactive Endocannabinoid System in Human Obesity and Type 2 Diabetes 2-AG: 2-arachidonoylglycerol BED: binge-eating disorder Adapted from Matias I et al. J Clin Endocrinol Metab 2006; 91: 3171-80 Reproduced with permission ** p≤0.01 vs. visceral fat from normoweight volunteers ## p≤0.01 vs. visceral fat from obese patients Fat of obese patients Blood of type 2 diabetic patients ** p≤0.01 vs. controls *** p≤0.005 vs. controls p<0.005 Anandamide (pmol/ml) Blood of obese women From Monteleone P et al. Neuropsychopharmacology 2005; 30: 1216-21 Reproduced with permission

6. Metabolic Effects of CB1 Blockade in Mice with Diet-Induced Obesity Adapted from Ravinet Trillou C et al. Am J Physiol Integr Comp Physiol 2003; 284: R345-5 Reproduced with permission 10 mg/kg/day for five weeks in HFD Mice STD-veh: standard diet vehicle HFD-veh: high fat diet vehicle HFD-SR10 mg: high fat diet + rimonabant 10 mg * p<0.001 vs. STD-veh ** p<0.001 vs. HFD-veh 2.0 1.6 1.2 0.8 0.4 0.0 * **

7. Metabolic Effects of CB1 Blockade in Mice with Diet-Induced Obesity Adapted from Poirier B et al. Diabetes Obes Metab 2005; 7: 65-72 0 24 (Weeks) 0 24 34 Standard diet (STD) High fat diet (HFD) STD Switched to STD HFD + Vehicle HFD + Rimonabant Lipoprotein cholesterol distribution in obese mice following a 10-week treatment with rimonabant HFD + Rimonabant Switched to STD vehicle STD vehicle HFD vehicle * p<0.001 vs. other groups † p<0.01 vs. STD groups ‡ p<0.05 vs. STD vehicle group 50 40 30 20 grams Body weight ‡ * 0 2 4 6 HDL-C † † mmol/l 6 6 8 8 10 10 12 12 HDL/LDL * ‡ 0.00 0.25 0.50 0.75 1.00 LDL-C TG mmol/l * * HDL-C: HDL cholesterol HDL/LDL: HDL cholesterol/LDL cholesterol LDL-C: LDL cholesterol TG: triglycerides