Inhibitors, Agonists, Screening Libraries
https://www.doczj.com/doc/e92301046.html, Data Sheet
BIOLOGICAL ACTIVITY:
URB597 is a potent, orally bioavailable FAAH inhibitor with IC50 of 4.6 nM, with no activity on other cannabinoid–related targets.IC50 value: 4.6 nM [1]
Target: FAAH
in vitro: URB597 binds in the hydrophobic pocket and catalytic core of FAAH that connects the active site residues to the membrane surface of FAAH [1]. URB597 reduces the expression of the LPS–induced enzymes cyclo–oxygenase 2 (COX–2) and inducible nitric oxide synthase (iNOS; NOS2) in primary rat microglial cell, with a concomitant reduction in the release of the inflammatory mediators prostaglandin E2 (PGE2) and (NO) nitric oxide [2].
in vivo: URB597 inhibits [3H]anandamide hydrolysis in rat brain membranes with a parallel increase in brain anandamide, OEA, and PEA content by inhibition of FAAH. URB597 enhances the hypothermia effect induced by ethanolamide by inhibiting FAAH [3]. When delivered intraperitonealy (0.3 mg/kg) URB597 reduces allodynia and hyperalgesia through cannabinoid CB1 and CB2
receptor–mediated analgesia in rats with inflammatory pain [4].
PROTOCOL (Extracted from published papers and Only for reference)
Enzyme assay [3]
FAAH–/– mice were anesthetized with halothane and sacrificed by decapitation. The brain and duodenum were removed and
homogenized in ice–cold Tris–HCL (20 mM, 10 vol, pH 7.4) containing 0.32 M sucrose. The homogenates were centrifuged at 800g for 15 min and then at 27,000g for 30 min. The 27,000g pellet was suspended in phosphate–buffered saline (pH = 7.4) and used for FAAH assays. To isolate the acid amidase activity, URB–597 was first subjected to two cycles of freezing and thawing and then centrifuged at 105,000 g for 60 min (Ueda et al., 2001). Acid amidase activity was analyzed in the supernatant, as described below. We measured FAAH activity at 37°C for 30 min in 0.5 ml of Tris buffer (50 mM, pH 7.5) containing fatty acid–free bovine serum albumin (0.05%),membrane protein (50 μg) and anandamide[ethanolamine–3H] (10,000 dpm, specific activity 20 Ci/mmol; American Radiolabeled Chemicals, St. Louis, MO). We quantified acid amidase activity at 37°C for 30 min in 0.5 ml of sodium phosphate buffer (50 mM, pH 5.0) containing 3 mM dithiothreitol, 0.1% Triton X–100, protein (50 μg), and palmitoyl[ethanolamine–3H] (30,000 dpm, specific activity 20 Ci/mmol; American Radiolabeled Chemicals). After stopping the reactions with chloroform/methanol (1:1, 1 ml), we measured radioactivity in the aqueous layers by liquid scintillation counting.
References:
[1]. Mor M, et al. Cyclohexylcarbamic acid 3'– or 4'–substituted biphenyl–3–yl esters as fatty acid amide hydrolase inhibitors: synthesis, quantitative structure–activity relationships, and molecular modeling studies. J Med Chem, 2004, 47(21), 4998–5008.
[2]. Tham CS, et al. Inhibition of microglial fatty acid amide hydrolase modulates LPS stimulated release of inflammatory mediators. FEBS Lett, 2007, 581(16),
Product Name:
URB–597Cat. No.:
HY-10864CAS No.:
546141-08-6Molecular Formula:
C 20H 22N 2O 3Molecular Weight:
338.40Target:
FAAH; FAAH; Autophagy Pathway:
Neuronal Signaling; Metabolic Enzyme/Protease; Autophagy Solubility:
10 mM in DMSO
2899–2904.
[3]. Fegley D, et al. Characterization of the fatty acid amide hydrolase inhibitor cyclohexyl carbamic acid 3'–carbamoyl–biphenyl–3–yl ester (URB597): effects on anandamide and oleoylethanolamide deactivation. J Pharmacol Exp Ther. 2005 Apr;313(1):352–8.
[4]. Jayamanne A, et al. Actions of the FAAH inhibitor URB597 in neuropathic and inflammatory chronic pain models. Br J Pharmacol, 2006, 147(3), 281–288.
Caution: Product has not been fully validated for medical applications. For research use only.
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