Pharmacological characterization of endocannabinoid transport and fatty acid amide hydrolase inhibitors.

@article{citeulike:1248922, abstract = {: 1. The mechanism of anandamide uptake and disposal has been an issue of considerable debate in the cannabinoid field. Several compounds have been reported to inhibit anandamide uptake or fatty acid amide hydrolase (FAAH; the primary catabolic enzyme of anandamide) activity with varying degrees of potency and selectivity. We recently reported the first evidence of a binding site involved in the uptake of endocannabinoids that is independent from FAAH. There are no direct comparisons of purported selective inhibitory compounds in common assay conditions measuring anandamide uptake, FAAH activity and binding activity. 2. A subset of compounds reported in the literature were tested in our laboratory under common assay conditions to measure their ability to (a) inhibit [(14)C]-anandamide uptake in cells containing (RBL-2H3) or cells lacking (HeLa) FAAH, (b) inhibit purified FAAH hydrolytic activity, and (c) inhibit binding to a putative binding site involved in endocannabinoid transport in both RBL and HeLa cell membranes. 3. Under these conditions, nearly all compounds tested inhibited (a) uptake of [(14)C]-anandamide, (b) enzyme activity in purified FAAH preparations, and (c) radioligand binding of [(3)H]-LY2183240 in RBL and HeLa plasma membrane preparations. General rank order potency was preserved within the three assays. However, concentration response curves were right-shifted for functional [(14)C]-anandamide uptake in HeLa (FAAH(-/-)) cells. 4. A more direct comparison of multiple inhibitors could be made in these three assay systems performed in the same laboratory, revealing more information about the selectivity of these compounds and the relationship between the putative endocannabinoid transport protein and FAAH. At least two separate proteins appear to be involved in uptake and degradation of anandamide. The most potent inhibitory compounds were right-shifted when transport was measured in HeLa (FAAH(-/-)) cells suggesting a requirement for a direct interaction with the FAAH protein to maintain high affinity binding of anandamide or inhibitors to the putative anandamide transport protein.}, address = {Eli Lilly \& Company, Lilly Research Laboratories, Lilly Corporate Center, Indianapolis, Indiana 46285, USA.}, author = {Dickason-Chesterfield, A. K. and Kidd, S. R. and Moore, S. A. and Schaus, J. M. and Liu, B. and Nomikos, G. G. and Felder, C. C. }, citeulike-article-id = {1248922}, doi = {http://dx.doi.org/10.1007/s10571-006-9072-6}, issn = {0272-4340}, journal = {Cell Mol Neurobiol}, keywords = {faah, inhibition, transport}, number = {4-6}, pages = {407--423}, posted-at = {2007-04-25 01:16:17}, priority = {2}, title = {Pharmacological characterization of endocannabinoid transport and fatty acid amide hydrolase inhibitors.}, url = {http://dx.doi.org/10.1007/s10571-006-9072-6}, volume = {26}, year = {2006} }

TY - JOUR ID - citeulike:1248922 L3 - citeulike-article-id:1248922 TI - Pharmacological characterization of endocannabinoid transport and fatty acid amide hydrolase inhibitors. JF - Cell Mol Neurobiol VL - 26 IS - 4-6 SP - 407 EP - 423 AD - Eli Lilly & Company, Lilly Research Laboratories, Lilly Corporate Center, Indianapolis, Indiana 46285, USA. SN - 0272-4340 N2 - : 1. The mechanism of anandamide uptake and disposal has been an issue of considerable debate in the cannabinoid field. Several compounds have been reported to inhibit anandamide uptake or fatty acid amide hydrolase (FAAH; the primary catabolic enzyme of anandamide) activity with varying degrees of potency and selectivity. We recently reported the first evidence of a binding site involved in the uptake of endocannabinoids that is independent from FAAH. There are no direct comparisons of purported selective inhibitory compounds in common assay conditions measuring anandamide uptake, FAAH activity and binding activity. 2. A subset of compounds reported in the literature were tested in our laboratory under common assay conditions to measure their ability to (a) inhibit [(14)C]-anandamide uptake in cells containing (RBL-2H3) or cells lacking (HeLa) FAAH, (b) inhibit purified FAAH hydrolytic activity, and (c) inhibit binding to a putative binding site involved in endocannabinoid transport in both RBL and HeLa cell membranes. 3. Under these conditions, nearly all compounds tested inhibited (a) uptake of [(14)C]-anandamide, (b) enzyme activity in purified FAAH preparations, and (c) radioligand binding of [(3)H]-LY2183240 in RBL and HeLa plasma membrane preparations. General rank order potency was preserved within the three assays. However, concentration response curves were right-shifted for functional [(14)C]-anandamide uptake in HeLa (FAAH(-/-)) cells. 4. A more direct comparison of multiple inhibitors could be made in these three assay systems performed in the same laboratory, revealing more information about the selectivity of these compounds and the relationship between the putative endocannabinoid transport protein and FAAH. At least two separate proteins appear to be involved in uptake and degradation of anandamide. The most potent inhibitory compounds were right-shifted when transport was measured in HeLa (FAAH(-/-)) cells suggesting a requirement for a direct interaction with the FAAH protein to maintain high affinity binding of anandamide or inhibitors to the putative anandamide transport protein. KW - faah KW - inhibition KW - transport AU - Dickason-Chesterfield, AK AU - Kidd, SR AU - Moore, SA AU - Schaus, JM AU - Liu, B AU - Nomikos, GG AU - Felder, CC PY - 2006///g UR - http://dx.doi.org/10.1007/s10571-006-9072-6 ER -