Deutsch D G, Lin S, Hill W A, Morse K L, Salehani D, Arreaza G, Omeir R L, Makriyannis A
Department of Biochemistry and Cell Biology, State University of New York at Stony Brook 11794, USA.
Biochem Biophys Res Commun. 1997 Feb 3;231(1):217-21. doi: 10.1006/bbrc.1997.6072.
Arachidonoyl ethanolamide (anandamide) is an endogenous ligand for cannabinoid receptors (CB1, CB2) and a putative neurotransmitter. Phenylmethylsulfonyl fluoride (PMSF) is an inhibitor of the enzyme (an amidase) which hydrolyzes anandamide to arachidonic acid and ethanolamine. We report here that fatty acid sulfonyl fluorides are potent inhibitors of anandamide metabolism. In order to investigate the SAR of these anandamide amidase inhibitors we tested a series of fatty acid (C12 to C20) sulfonyl fluorides both as inhibitors of anandamide degradation and as ligands for the central cannabinoid receptor (CB1). AM374 (palmitylsulfonyl fluoride, C16) was approximately 20 times more potent than PMSF and 50 times more potent than arachidonyltrifluoromethyl ketone in preventing the hydrolysis of anandamide in brain homogenates. AM374 was over a thousand-fold more effective than PMSF in inhibiting the amidase in cultured cells. The C12 to C18 sulfonyl fluoride analogs were equipotent as inhibitors of the amidase and the reverse reaction (the synthase) with nanomolar IC50 values. These compounds generally showed decreasing affinity for the CB1 receptor as the chain length increased; thus, C12 sulfonylfluoride had an IC50 of 18 nM and C20 sulfonylfluoride had an IC50 of 78 microM. The C14, C16, and C18 sulfonyl fluorides showed high selectivity for the amidase over the CB1 receptor and thus are potentially useful selective anandamide amidase inhibitors.
花生四烯酸乙醇酰胺（阿南达米德）是大麻素受体（CB1、CB2）的内源性配体，也是一种假定的神经递质。苯甲基磺酰氟（PMSF）是一种酶（酰胺酶）的抑制剂，该酶可将阿南达米德水解为花生四烯酸和乙醇胺。我们在此报告，脂肪酸磺酰氟是阿南达米德代谢的有效抑制剂。为了研究这些阿南达米德酰胺酶抑制剂的构效关系，我们测试了一系列脂肪酸（C12至C20）磺酰氟，它们既是阿南达米德降解的抑制剂，也是中枢大麻素受体（CB1）的配体。在防止脑匀浆中阿南达米德水解方面，AM374（棕榈酰磺酰氟，C16）的效力比PMSF高约20倍，比花生四烯酰三氟甲基酮高50倍。在抑制培养细胞中的酰胺酶方面，AM374比PMSF有效一千多倍。C12至C18磺酰氟类似物作为酰胺酶抑制剂和逆反应（合酶）的抑制剂具有同等效力，IC50值为纳摩尔级。随着链长增加，这些化合物对CB1受体的亲和力通常会降低；因此，C12磺酰氟的IC50为18 nM，C20磺酰氟的IC50为78 μM。C14、C16和C18磺酰氟对酰胺酶的选择性高于CB1受体，因此可能是有用的选择性阿南达米德酰胺酶抑制剂。
