Chemical probes to potently and selectively inhibit endocannabinoid cellular reuptake

The extracellular effects of the endocannabinoids anandamide and 2-arachidonoyl glycerol are terminated by enzymatic hydrolysis after crossing cellular membranes by facilitated diffusion. The lack of potent and selective inhibitors for endocannabinoid transport has prevented the molecular characteri...

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Published inProceedings of the National Academy of Sciences - PNAS Vol. 114; no. 25; pp. E5006 - E5015
Main Authors Chicca, Andrea, Nicolussi, Simon, Bartholomäus, Ruben, Blunder, Martina, Rey, Alejandro Aparisi, Petrucci, Vanessa, del Carmen Reynoso-Moreno, Ines, Viveros-Paredes, Juan Manuel, Gens, Marianela Dalghi, Lutz, Beat, Schiöth, Helgi B., Soeberdt, Michael, Abels, Christoph, Charles, Roch-Philippe, Altmann, Karl-Heinz, Gertsch, Jürg
Format Journal Article
LanguageEnglish
Published United States National Academy of Sciences 20.06.2017
SeriesPNAS Plus
Subjects
2-AG
Analgesics
Anandamide
Animals
Anti-Anxiety Agents - pharmacology
Anti-Inflammatory Agents - pharmacology
Arachidonic Acids - metabolism
Biological Sciences
Biological Transport - drug effects
Brain
Brain - drug effects
Brain - metabolism
Cannabinoid receptors
Cannabinoids
Cell Line, Tumor
Cell Membrane - drug effects
Cell Membrane - metabolism
Chemical synthesis
Chemicals
Diffusion
endocannabinoid reuptake
endocannabinoid system
Endocannabinoids - metabolism
Enzymes
Exploitation
Glycerides - metabolism
Glycerol
Humans
Hydrolysis - drug effects
inhibitor
Inhibitors
lipid transport
Membrane trafficking
Membranes
Mice
Mice, Inbred BALB C
Mice, Inbred C57BL
Pharmacology
Photoactivation
PNAS Plus
Polyunsaturated Alkamides - metabolism
Probes
Proteins
Receptors, Cannabinoid - metabolism
U937 Cells
endocannabinoid reuptake
lipid transport
endocannabinoid system
inhibitor
2-AG
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Summary:The extracellular effects of the endocannabinoids anandamide and 2-arachidonoyl glycerol are terminated by enzymatic hydrolysis after crossing cellular membranes by facilitated diffusion. The lack of potent and selective inhibitors for endocannabinoid transport has prevented the molecular characterization of this process, thus hindering its biochemical investigation and pharmacological exploitation. Here, we report the design, chemical synthesis, and biological profiling of natural product-derived N-substituted 2,4-dodecadienamides as a selective endocannabinoid uptake inhibitor. The highly potent (IC50 = 10 nM) inhibitor N-(3,4-dimethoxyphenyl)ethyl amide (WOBE437) exerted pronounced cannabinoid receptor-dependent anxiolytic, antiinflammatory, and analgesic effects in mice by increasing endocannabinoid levels. A tailored WOBE437-derived diazirine-containing photoaffinity probe (RX-055) irreversibly blocked membrane transport of both endocannabinoids, providing mechanistic insights into this complex process. Moreover, RX-055 exerted site-specific anxiolytic effects on in situ photoactivation in the brain. This study describes suitable inhibitors to target endocannabinoid membrane trafficking and uncovers an alternative endocannabinoid pharmacology.
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1A.C. and S.N. contributed equally to this work.
Author contributions: A.C., S.N., M.B., B.L., K.-H.A., and J.G. designed research; A.C., S.N., R.B., M.B., A.A.R., V.P., I.d.C.R.-M., J.M.V.-P., and M.D.G. performed research; M.S., C.A., and R.-P.C. contributed new reagents/analytic tools; A.C., S.N., R.B., M.B., A.A.R., V.P., I.d.C.R.-M., J.M.V.-P., B.L., H.B.S., M.S., and J.G. analyzed data; and A.C., S.N., M.S., K.-H.A., and J.G. wrote the paper.
Edited by Benjamin F. Cravatt, The Scripps Research Institute, La Jolla, CA, and approved May 10, 2017 (received for review March 14, 2017)
ISSN:0027-8424
1091-6490
1091-6490
DOI:10.1073/pnas.1704065114