N-arachidonoyl glycine (NAGly, Tocris Cookson, Bristol, UK), 1,3-Dimethoxy-5-methyl-2-[(1R,6R)-3-methyl-6-(1-methylethenyl)-2-cyclohexen-1-yl]benzene (O-1918, Tocris) and 6-Iodo-2-methyl-1-[2-(4-morpholinyl)ethyl]-1H-indol-3-yl](4-methoxyphenyl) methanone (AM630, Tocris) were dissolved in DMSO. Lipopolysaccharide (LPS, Sigma Aldrich, Munich, Germany), N-Methyl-D-Aspartate (NMDA, Sigma Aldrich) and adenosine triphosphate (ATP, Sigma Aldrich) were diluted in Aqua dest. All chemicals were stored at −20 °C or −80 °C until use.
O 1918
Manufactured by Bio-Techne
Sourced in United Kingdom
O-1918 is a synthetic cannabinoid receptor antagonist. It acts as an inverse agonist of the CB1 receptor, which is involved in the regulation of various physiological processes. The core function of O-1918 is to modulate the activity of the endocannabinoid system.
Automatically generated - may contain errors
Lab products found in correlation
Am251, by Bio-Techne (4 mentions)
Am630, by Bio-Techne (4 mentions)
Abn cbd, by Bio-Techne (2 mentions)
Gw9662, by Bio-Techne (2 mentions)
Gw6471, by Bio-Techne (2 mentions)
Opti mem, by Thermo Fisher Scientific (2 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions)
N arachidonoyl glycine nagly, by Bio-Techne (1 mentions)
N methyl d aspartate nmda, by Merck Group (1 mentions)
Adenosine triphosphate (atp), by Merck Group (1 mentions)
Lipopolysaccharide lps, by Merck Group (1 mentions)
Trinitrobenzene sulfonic acid tnbs, by Merck Group (1 mentions)
Capsazepine, by Bio-Techne (1 mentions)
Urb597, by Merck Group (1 mentions)
Cgrp8 37, by Bio-Techne (1 mentions)
Tram 34, by Bio-Techne (1 mentions)
U46619, by Bio-Techne (1 mentions)
Cay10441, by Cayman Chemical (1 mentions)
L161982, by Bio-Techne (1 mentions)
Pentobarbitone sodium, by Biowet (1 mentions)
10 protocols using o 1918
1
Pharmacological Agents in Cell Assays
2
Pharmacological Modulation of TNBS Colitis
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Trinitrobenzene sulfonic acid (TNBS) was purchased from Sigma-Aldrich (Oakville, Ontario, Canada). Abn-CBD, dissolved in methyl acetate, O-1918 (1,3-Dimethoxy-5-methyl-2-[(1R,6R)-3-methyl-6-(1-methylethenyl)-2-cyclohexen-1-yl]benz-ene), AM630 (6-iodo-2-methyl-1-[2-(4-morpholinyl)ethyl-1H-indol-3-yl](4-methoxyphenyl)methanone) and AM251 (N-(Piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide) were obtained from Tocris Bioscience (Bristol, UK). Because of its toxicity, methyl acetate was evaporated prior to the in vivo experiments and ethanol was used instead as a solvent. Abn-CBD was then further diluted in Tween 80 (10 %) and sterile saline. Vehicle consisted of ethanol, Tween 80 and sterile saline (1:1:8). AM630 and AM251 were dissolved in dimethyl sulfoxide (DMSO, 99.7 %) and further diluted with vehicle. 45 min prior to the induction of TNBS colitis, mice were injected intraperitoneally (i.p.) with 5 mg/kg AM630, AM251, O-1918 or vehicle, followed by 5 mg/kg Abn-CBD, 15 min later. As a single dose of Abn-CBD was ineffective (preliminary data not shown), mice were injected twice daily for 3 days. For in vitro assays, 10 mM stock solutions (in DMSO) of Abn-CBD and the CB receptor antagonists were prepared.
3
GPR52 Receptor Docking Protocol
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Schrodinger 2020-3 release was used to perform the docking experiments presented here. The GPR52 publicly released structure [18 (link)] (pdb code 6LI0) was prepared with the Protein Preparation Wizard. Glide module was used to perform the dockings with SP precision and extended sampling. Dockings were done with and without water 304 from the 6LI0 structure to compare poses that could interact with that water or displace it.
O-1918 and CBD are commercially available from Tocris (2288 and 1570 respectively).
Compound 1 and Compound 2 syntheses are reported in the literature [19 (link)].
O-1918 and CBD are commercially available from Tocris (2288 and 1570 respectively).
Compound 1 and Compound 2 syntheses are reported in the literature [19 (link)].
4
Endocannabinoid Receptor Ligand Preparation
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All endocannabinoids were purchased from Tocris (Bristol, UK) and dissolved in ethanol to a stock concentration of 10 mM, except 2-AG which was purchased from Abcam (Cambridge, UK) and dissolved in acetonitrile. AM251, AM630, GW6471, GW9662 (all 100 nM), capsazepine, O-1918 (both 1 μM) (all dissolved in dimethyl sulfoxide) and CGRP8–37 (2 μM, dissolved in distilled water) were all purchased from Tocris and URB597 (1 μM, dissolved in dimethyl sulfoxide) was purchased from Sigma (Dorset, UK). All were dissolved to a stock solution of 10 mM.
5
Preparation and Use of Chemical Compounds in Biomedical Research
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Cremophor EL, DOCA, dimethyl sulfoxide (DMSO), N,N-dimethylformamide (DMF) and Tween-80; pentobarbitone sodium (Biowet, Puławy, Poland) and sodium chloride (NaCl) (Chempur, Piekary Śląskie, Poland).
Unless otherwise stated, all chemicals were purchased from Sigma-Aldrich (Munich, Germany). U46619, CBD, GW9662, L161982, O-1918, TRAM-34, AM251, and AM630 were bought from Tocris Bioscience (Bristol, UK). Cay10441 was purchased from Cayman Chemical Co. (Ann Arbor, Michigan, USA). Stock solutions of CBD, U46619, GW9662, AM251, O-1918, capsazepine, nimesulide, Cay10441 and RN1734 were made to 10 mmol/l in ethanol; AM630, L161982, UCL1684 and TRAM-34 were made to 10 mmol/l in dimethylsulfoxide. Their final concentrations were prepared by dilutions with deionized water, which adjusted the final concentrations of ethanol or DMSO 0.1% v/v or less with the exception of RN1734 (ethanol final concentration 0.2% v/v). Stock solutions of l -NAME (300 mmol/l), phenylephrine and acetylcholine (all 10 mmol/l) were made in distilled water. Indomethacin was dissolved in 0.5 mol/l NaHCO3 and DETCA directly into Krebs.
Unless otherwise stated, all chemicals were purchased from Sigma-Aldrich (Munich, Germany). U46619, CBD, GW9662, L161982, O-1918, TRAM-34, AM251, and AM630 were bought from Tocris Bioscience (Bristol, UK). Cay10441 was purchased from Cayman Chemical Co. (Ann Arbor, Michigan, USA). Stock solutions of CBD, U46619, GW9662, AM251, O-1918, capsazepine, nimesulide, Cay10441 and RN1734 were made to 10 mmol/l in ethanol; AM630, L161982, UCL1684 and TRAM-34 were made to 10 mmol/l in dimethylsulfoxide. Their final concentrations were prepared by dilutions with deionized water, which adjusted the final concentrations of ethanol or DMSO 0.1% v/v or less with the exception of RN1734 (ethanol final concentration 0.2% v/v). Stock solutions of
6
Characterization of Human GPR18 Ligands
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In this study, three selective ligands of human GPR18 were used, including two agonists with undisclosed structure (PSB-MZ-1415, EC50 0.019 µM and PSB-MZ-1440, EC50 0.061 µM; chemical details will be shortly published elsewhere) and the antagonist (Z)-2-(3-(6-(4-chlorophenoxy)hexyloxy)benzylidene)-6,7-dihydro-2H-imidazo[2,1-b][1,3]thiazin-3(5H)-one (PSB-CB-27, IC50 0.65 µM). The three ligands were synthetized at the Department of Technology and Biotechnology of Drugs, Faculty of Pharmacy, Jagiellonian University, Kraków, Poland, and identified and characterized in the Department of Pharmaceutical & Medicinal Chemistry, University of Bonn, Germany ([33 (link)] and unpublished results).
U46619 ((5Z)-7-{(1R,4S,5S,6R)-6-[(1E,3S)-3-hydroxyoct-1-en-1-yl]-2-oxabicyclo[2.2.1]-heptan-5-yl}hept-5-enoic acid), O-1918 (1,3-dimethoxy-5-methyl-2-[(1R,6R)-3-methyl-6-prop-1-en-2-ylcyclohex-2-en-1-yl]benzene), NAGly, and Abn-CBD were purchased from Tocris Bioscience (Bristol, UK). Stock solutions of these substances were made to 10 mM in ethanol; their final concentrations were prepared by dilutions with deionized water, which led to final concentrations of ethanol of <0.7% v/v. Stock solutions of PSB-MZ-1415, PSB-MZ-1440, and PSB-CB-27 were made only to 10 mM in dimethyl sulfoxide (DMSO). The final concentration of DMSO was <1.0% v/v.
U46619 ((5Z)-7-{(1R,4S,5S,6R)-6-[(1E,3S)-3-hydroxyoct-1-en-1-yl]-2-oxabicyclo[2.2.1]-heptan-5-yl}hept-5-enoic acid), O-1918 (1,3-dimethoxy-5-methyl-2-[(1R,6R)-3-methyl-6-prop-1-en-2-ylcyclohex-2-en-1-yl]benzene), NAGly, and Abn-CBD were purchased from Tocris Bioscience (Bristol, UK). Stock solutions of these substances were made to 10 mM in ethanol; their final concentrations were prepared by dilutions with deionized water, which led to final concentrations of ethanol of <0.7% v/v. Stock solutions of PSB-MZ-1415, PSB-MZ-1440, and PSB-CB-27 were made only to 10 mM in dimethyl sulfoxide (DMSO). The final concentration of DMSO was <1.0% v/v.
7
Investigating Cannabinoid Receptor Signaling
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CBD was bought from Biomol GmbH (Hamburg, Germany). JWH-133 and THC were bought from Bio-Techne GmbH (Wiesbaden, Germany) and Lipomed GmbH (Herne, Germany), respectively. R(+)-methanandamide, capsazepine and NAC were obtained from Sigma-Aldrich (Taufkirchen, Germany). AM251 and AM630 were obtained from Biozol (Eching, Germany). O-1602 and O-1918 were purchased from Tocris Bioscience (Bristol, UK) and Cayman Chemicals (Ann Arbor, Michigan, USA), respectively. SnPPIX was obtained from Enzo Life Sciences (Lörrach, Germany). The transfection reagent Lipofectamine™ RNAiMAX, OptiMEM and hPDGF-BB were purchased from Thermo Fisher Scientific Inc. (Schwerte, Germany). siRNA targeting HO-1 was purchased from Santa Cruz Biotechnology (Heidelberg, Germany; sc-35554). Negative control siRNA was from Qiagen (Hilden, Germany; cat. no. 1022076). Accutase cell detachment solution was obtained from Merck Chemicals GmbH (Darmstadt, Germany).
8
Cannabinoid Compounds Comparison
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Δ9-THC and CBD were obtained through the National Institutes on Drug Abuse drug supply program. O-1918 and Tocrisolve were obtained from Tocris Bioscience.
9
Cannabinoid Receptor Ligand Preparation
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CBG and CBDV were kindly gifted by STI pharmaceuticals. Both compounds were dissolved in 100% ethanol to 10 mM and were stored at −20°C. AM251, AM630, GW6471, GW9962, O1918, CID16020046, SB366791 (Tocris, United Kingdom) were dissolved in dimethyl sulfoxide as stock solutions of 10 mM. (S)-WAY100135 was dissolved in deionized water. Antagonists were stored at −20°C and dilutions were made fresh as required.
10
GPR18 Modulation in CYP-Induced Cystitis
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To analyse the role of GPR18 in CYP-induced cystitis, we used a GPR18 agonist, RvD2 (MCE, USA), and a GPR18 antagonist, O-1918 (Tocris, UK). RvD2 was dissolved in sterile PBS while minimizing exposure to light. RvD2 (10 ng, 100 ng) or vehicle (2% ethanol in PBS) was injected intrathecally as described in previous studies.18 (link),19 (link) Before using O-1918, the methyl acetate was evaporated under a gentle stream of nitrogen, and vehicle was immediately added. The final concentration of O-1918 was 25 mM. Spinal puncture was performed between the L5 and L6 levels using a PE-10 catheter for intrathecal injection of the reagent (30 µL).
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