Methyl arachidonyl fluorophosphonate: A potent irreversible inhibitor of anandamide amidase

被引:207
作者
Deutsch, DG
Omeir, R
Arreaza, G
Salehani, D
Prestwich, GD
Huang, Z
Howlett, A
机构
[1] SUNY STONY BROOK,INST CELL & DEV BIOL,STONY BROOK,NY 11794
[2] MERCK FROSST CTR THERAPEUT RES,DEPT BIOCHEM & MOL BIOL,POINTE CLAIRE,PQ H9R 4P8,CANADA
[3] ST LOUIS UNIV,SCH MED,DEPT PHARMACOL & PHYSIOL SCI,ST LOUIS,MO 63104
关键词
anandamide; anandamide amidase; amidohydrolase; cannabinoid receptor; methyl arachidonyl fluorophosphonate; sleep-inducing factor;
D O I
10.1016/S0006-2952(96)00830-1
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
Anandamide amidase (EC 3.5.1.4) is responsible for tie hydrolysis of arachidonoyl ethanolamide (anandamide). Relatively selective and potent enzyme reversible inhibitors effective in the low micromolar range, such as arachidonyl trifluoromethyl ketone (Arach-CF3), have been described (Koutek et al., J Biol Chem 269: 22937-22940, 1994). In the current study, methyl arachidonyl fluorophosphonate (MAFP), an arachidonyl binding site directed phosphonylation reagent, was tested as an inhibitor of anandamide amidase and as a ligand for the CB, cannabinoid receptor. MAFP was 800 times more potent than Arach-CF3 and phenylmethylsulfonyl fluoride (PMSF) as an amidase inhibitor in rat brain homogenates. In intact neuroblastoma cells, MAFP was also approximately 1000-fold more potent than Arach-CF3. MAFP demonstrated selectivity towards anandamide amidase for which it was approximately 3000 and 30,000-fold more potent than it was towards chymotrypsin and trypsin, respectively. MAFP displaced [H-3]CP-55940 binding to the CB1 cannabinoid receptor with an IC50 of 20 nM vs 40 nM for anandamide. It bound irreversibly and prevented subsequent binding of the cannabinoid radioligand [H-3]CP-55940 at that locus. These studies suggest that MAFP is a potent and specific inhibitor of anandamide amidase and, in addition, can interact with the cannabinoid receptors at the cannabinoid binding site. This is the first report of a potent and relatively selective irreversible inhibitor of arachidonoyl ethanolamide amidase.
引用
收藏
页码:255 / 260
页数:6
相关论文
共 18 条
  • [1] (R)-METHANANDAMIDE - A CHIRAL NOVEL ANANDAMIDE POSSESSING HIGHER POTENCY AND METABOLIC STABILITY
    ABADJI, V
    LIN, SY
    TAHA, G
    GRIFFIN, G
    STEVENSON, LA
    PERTWEE, RG
    MAKRIYANNIS, A
    [J]. JOURNAL OF MEDICINAL CHEMISTRY, 1994, 37 (12) : 1889 - 1893
  • [2] PHARMACOLOGICAL AND BEHAVIORAL-EVALUATION OF ALKYLATED ANANDAMIDE ANALOGS
    ADAMS, IB
    RYAN, W
    SINGER, M
    RAZDAN, RK
    COMPTON, DR
    MARTIN, BR
    [J]. LIFE SCIENCES, 1995, 56 (23-24) : 2041 - 2048
  • [3] EFFECTS OF ANANDAMIDE ON CANNABINOID RECEPTORS IN RAT-BRAIN MEMBRANES
    CHILDERS, SR
    SEXTON, T
    ROY, MB
    [J]. BIOCHEMICAL PHARMACOLOGY, 1994, 47 (04) : 711 - 715
  • [4] CHEMICAL CHARACTERIZATION OF A FAMILY OF BRAIN LIPIDS THAT INDUCE SLEEP
    CRAVATT, BF
    PROSPEROGARCIA, O
    SIUZDAK, G
    GILULA, NB
    HENRIKSEN, SJ
    BOGER, DL
    LERNER, RA
    [J]. SCIENCE, 1995, 268 (5216) : 1506 - 1509
  • [5] ANANDAMIDE AMIDOHYDROLASE ACTIVITY IN RAT-BRAIN MICROSOMES - IDENTIFICATION AND PARTIAL CHARACTERIZATION
    DESARNAUD, F
    CADAS, H
    PIOMELLI, D
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (11) : 6030 - 6035
  • [6] ENZYMATIC-SYNTHESIS AND DEGRADATION OF ANANDAMIDE, A CANNABINOID RECEPTOR AGONIST
    DEUTSCH, DG
    CHIN, SA
    [J]. BIOCHEMICAL PHARMACOLOGY, 1993, 46 (05) : 791 - 796
  • [7] FORMATION AND INACTIVATION OF ENDOGENOUS CANNABINOID ANANDAMIDE IN CENTRAL NEURONS
    DIMARZO, V
    FONTANA, A
    CADAS, H
    SCHINELLI, S
    CIMINO, G
    SCHWARTZ, JC
    PIOMELLI, D
    [J]. NATURE, 1994, 372 (6507) : 686 - 691
  • [8] THE BINDING OF NOVEL PHENOLIC DERIVATIVES OF ANANDAMIDE TO BRAIN CANNABINOID RECEPTORS
    EDGEMOND, WS
    CAMPBELL, WB
    HILLARD, CJ
    [J]. PROSTAGLANDINS LEUKOTRIENES AND ESSENTIAL FATTY ACIDS, 1995, 52 (2-3): : 83 - 86
  • [9] HILLARD CJ, 1995, J NEUROCHEM, V64, P677
  • [10] Functional identification of the active-site nucleophile of the human 85-kDa cytosolic phospholipase A(2)
    Huang, Z
    Payette, P
    Abdullah, K
    Cromlish, WA
    Kennedy, BP
    [J]. BIOCHEMISTRY, 1996, 35 (12) : 3712 - 3721