Anandamide analogs as affinity probes for proteins of the endocannabinoid system

Date of Completion

January 2000

Keywords

Chemistry, Pharmaceutical

Degree

Ph.D.

Abstract

Anandamide, the major endogenous cannabimimetic, was used as a molecular template for the design and synthesis of several series of compounds. The objective of this study was to identify the structural determinants for recognition of endocannabinoid substances by each of the four proteins of the endocannabinoid system (CB1 and CB2 receptors, anandamide amidase and anandamide transporter protein). The information derived from this structure-activity relationship study was used to guide the design of pharmacological probes for these proteins. ^ A strategy was identified for the improvement of the metabolic stability of anandamide. First, introduction of methyl groups at the 1′ and 2′ positions of anandamide, led to increased resistance to anandamide amidase. 1′-methylation was proven to be superior in providing hydrolytic stability. The resultant methylated analogs exhibited a significant degree of stereoselectivity in their interactions with both, the CB1 receptor and anandamide amidase. Based on these results, a partial CB1 pharmacophoric model was proposed. ^ The structure-activity relationship that was developed for anandamide amidase allowed the design of a ligand affinity chromatography system for the purification of this enzyme. This chromatography system made possible a 4,500-fold purification of anandamide amidase. The pure enzyme was functional and had similar substrate specificity with that of the crude enzyme preparation. Furthermore, arachidonyl aldehyde and its congeners were identified as potent, tight binding, reversible inhibitors of anandamide amidase. ^

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