Morphine sulfate (National Institute on Drug Abuse Drug Supply Program, Bethesda, MD, USA) was dissolved in sterile water, phencyclidine hydrochloride (Sigma-Aldrich, St Louis, MO, USA) was dissolved in 0.9% saline, and 2-[(1R,2R,5R)-5-hydroxy-2-(3-hydroxypropyl) cyclohexyl]-5-(2-methyloctan-2-yl)phenol (CP55940; Sigma-Aldrich, St. Louis, MO, USA) was dissolved in pure ethanol (2 ml ethanol per bottle containing 10 mg CP55940) then diluted with a solution of 1:1:18 ethanol:emulphor:saline. Drugs were administered subcutaneously in a volume of 0.2 – 0.8 ml. Pretreatment times were 15 min for morphine and phencyclidine and 60 min for CP55940; behavioral effects of morphine and CP55940 are apparent with 15 and 60 min, respectively, and last at least 90 min in various operant tasks, including delay discounting (e.g., Maguire et al., 2012 (link), 2016 (link); Minervini and France, 2018 (link)). All doses were expressed as the weight of the forms noted above.
Cp55940
Manufactured by Merck Group
Sourced in United States
CP55940 is a synthetic cannabinoid receptor agonist. It functions as a potent and selective agonist for the CB1 and CB2 cannabinoid receptors.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Epinephrine, by Merck Group (2 mentions)
Sr144528, by Cayman Chemical (2 mentions)
Am281, by Bio-Techne (2 mentions)
Win55 212 2, by Cayman Chemical (2 mentions)
Zaprinast, by Merck Group (1 mentions)
Formic acid, by J&K Scientific (1 mentions)
Bradykinin, by Merck Group (1 mentions)
Histamine, by Merck Group (1 mentions)
Ham s f 12k medium, by Merck Group (1 mentions)
Mccoy s 5a medium, by Merck Group (1 mentions)
Dulbecco s modified eagle s medium dmem, by Merck Group (1 mentions)
Acetylcholine, by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Hplc grade acetonitrile, by Merck Group (1 mentions)
Epinephrine bitartrate, by Merck Group (1 mentions)
Rauwolscine, by Merck Group (1 mentions)
Zm241385, by Merck Group (1 mentions)
5 n ethylcarboxamidoadenosine neca, by Merck Group (1 mentions)
Dulbecco s modified eagle medium dmem buffer, by Thermo Fisher Scientific (1 mentions)
21 protocols using cp55940
1
Dissolving Drugs for Behavioral Effects
2
Subcutaneous administration of morphine and CP55940
3
Cannabinoid Effects on Visual Cortex Neuronal Activity
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CP55940 (Sigma) was dissolved in ethanol to achieve a stock solution of 3 mg/ml. For each animal, a 0.3 mg/kg dose of CP55940 injection solution was prepared using a mixture of 10% stock solution, 10% Cremaphor and 80% lactated Ringer’s solution. For vehicle-only injections, an equal volume of 10% ethanol, 10% Cremaphor and 80% lactated Ringer’s was prepared. Injections were delivered intravenously through either the cubital or femoral veins.
Baseline neuronal responses were initially recorded in response to the pseudorandom checkerboard stimulus. Following this, the vehicle-only injection was administered and after 15 minutes, responses to visual stimuli were again recorded. Afterwards, a 0.3 mg/kg dose of CP55940 was delivered and 15 minutes later, three recording sessions (spontaneous activity and receptive field mapping) were completed. Each recording session lasted 22 minutes. The baseline and vehicle-only recordings were pooled to form the “control condition.” Similarly, the three post-CP55940 infusion recordings were pooled to form the cannabinoid condition. Pooling over datasets helped reduce the effect of variability in neuronal responses.
Baseline neuronal responses were initially recorded in response to the pseudorandom checkerboard stimulus. Following this, the vehicle-only injection was administered and after 15 minutes, responses to visual stimuli were again recorded. Afterwards, a 0.3 mg/kg dose of CP55940 was delivered and 15 minutes later, three recording sessions (spontaneous activity and receptive field mapping) were completed. Each recording session lasted 22 minutes. The baseline and vehicle-only recordings were pooled to form the “control condition.” Similarly, the three post-CP55940 infusion recordings were pooled to form the cannabinoid condition. Pooling over datasets helped reduce the effect of variability in neuronal responses.
4
Natural Compound Library Preparation
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Natural compounds listed in Table S1 were purchased from Shanghai Yuanye Bio-technology (Shanghai, China), Shanghai Sidend Technical Service (Shanghai, China), National Institutes for Food and Drug Control (Beijing, China), Chengdu Profa Technology Development (Chengdu, China) and Chengdu Push Bio-technology (Chengdu, China). Zaprinast, epinephrine, histamine, bradykinin, CP55940 and acetylcholine were obtained from Sigma-Aldrich (St. Louis, MO, USA). Carbachol was obtained from Tocris Bioscience Co. (St. Louis, MO, USA). Hank's balanced salt solution (HBSS), HEPES, fetal bovine serum (FBS) were from Thermo Fisher Scientific (Waltham, MA). Ham's F12K medium, Dulbecco's modified Eagle's medium (DMEM) and McCoy's 5A medium were bought from Sigma-Aldrich (St Louis, MO, USA). HPLC grade acetonitrile and methanol were purchased from Merck (Darmstadt, Germany). Water was purified from a MilliQ water system (Billerica, MA, USA). Formic acid was obtained from J&K Scientific (Beijing, China). All natural compounds were prepared in 100 mM and were dissolved in 100 % dimethyl sulfoxide (DMSO) and diluted with the assay buffer (1 × Hank's balanced salt solution (HBSS) buffer, 10 mM HEPES, pH 7.4) to the desired concentrations before DMR assays.
5
Cell Lines and Reagents for Bioassays
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A transfected Chinese hamster ovary (CHO), Hela, a murine interleukin-3 dependent pro-B (Ba/F3), PC12, and rat basophil leukemia cell lines were obtained from Eurofins Scientific (Eurofins-Cerep, Le Bois I’Eveque, France). Buffers—Dulbecco’s modified Eagle medium (DMEM) buffer, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer, and Hank’s balanced salt solution (HBSS) buffer—were purchased from Invitrogen (Carlsbad, CA, USA). The reference agonists: 5′-N-ethylcarboxamidoadenosine (NECA), epinephrine bitartrate, epinephrine, DPDPE, CP 55940, linoleic acid, GLP-1(7–37, arginine vasopressin (AVP), and serotonin, and antagonists: ZM 241385, RX-821002, rauwolscine, naltriben mesylate, AM 281, exendin-3(9–39), [d(CH2)51, Tyr (Me)2]-AVP, (S)-WAY-100635, and GR55562) were obtained from Sigma-Aldrich (St. Louis, MO, USA). All other chemicals and reagents purchased from Merck and Fluka were of the highest available grade unless otherwise stated.
6
Cannabinoid Receptor Binding Assay
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192IgG-saporin was acquired from Millipore
(Temecula, CA, USA). [35S]GTPγS (1250 Ci/mmol) and
[3H]CP55,940 (131.8 Ci/mmol) were acquired from PerkinElmer
(Boston MA, USA). The [14C]-microscales, used as standards
in the autoradiographic experiments, were acquired from American Radiolabeled
Chemicals (St. Louis, MO, USA). Kodak Biomax MR β-radiation-sensitive
films, bovine serum albumin (BSA),dl -dithiothreitol, adenosine
deaminase, guanosine 5′-diphosphate, guanosine 5′-O-3-thiotriphosphate
(GTPγS), ketamine and xylazine, as well as CP55,940 were acquired
from Sigma-Aldrich (St. Louis, MO, USA). SR141716A and HU308 were
acquired from Tocris Bioscience (Bristol, UK) and SR144528 from Cayman-Chemicals
(MI, USA). All the compounds used were of the highest commercially
available quality.
(Temecula, CA, USA). [35S]GTPγS (1250 Ci/mmol) and
[3H]CP55,940 (131.8 Ci/mmol) were acquired from PerkinElmer
(Boston MA, USA). The [14C]-microscales, used as standards
in the autoradiographic experiments, were acquired from American Radiolabeled
Chemicals (St. Louis, MO, USA). Kodak Biomax MR β-radiation-sensitive
films, bovine serum albumin (BSA),
deaminase, guanosine 5′-diphosphate, guanosine 5′-O-3-thiotriphosphate
(GTPγS), ketamine and xylazine, as well as CP55,940 were acquired
from Sigma-Aldrich (St. Louis, MO, USA). SR141716A and HU308 were
acquired from Tocris Bioscience (Bristol, UK) and SR144528 from Cayman-Chemicals
(MI, USA). All the compounds used were of the highest commercially
available quality.
7
Osteoclastogenesis Assay Protocol
8
Synthesis and Characterization of Novel Cannabinoid Compounds
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Forskolin, WIN 55,212-2 and CP 55,940 (both CB1R/CB2R agonist), and fluoro-3 acetoxymethyl ester were purchased from Sigma–Aldrich Corp., (St. Louis, MO, United States). TXX-522 and SR141716A were synthesized and prepared by the New Drug Design Center of our Institute. Their purity and structure were confirmed using high-performance liquid chromatography, mass spectrometry, and proton (1H)-nuclear magnetic resonance. The structures of TXX-522 [(5-(4-chlorophenyl)-1-(2, 4-dichlorophenyl)-4-methyl-1H-pyrazol-3-yl) (piperidin-1-yl) methanone] and SR141716A are shown in Figure 1 .
9
Pharmacological Modulation of Membrane Permeability
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The following were used in the experiments: Evans Blue (Sigma Aldrich, St. Louis, MO, USA), WIN 55,212-2 (Cayman Chemicals, Ann Arbor, MI, USA); CP 55,940 (Sigma Aldrich); bethanechol (Sigma Aldrich), loperamide (Sigma Aldrich), JZL184 (Cayman Chemicals).
10
Pharmacological Evaluation of Remifentanil and Cannabinoids
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