The agonist and antagonist drugs and the dosage were selected according to the previous studies and on pilot experiments in our laboratory (21 (link)22 (link)23 ). For all tests, the CBHE (50, 150, and 300 mg/kg, i.p.) and cnicin (8, 20, and 30 mg/kg, i.p.) were dissolved in dimethyl sulfoxide (DMSO). Morphine sulfate (Morph; 1 mg/kg, i.p.), naloxone (NLX; 1 mg/kg, i.p.), diclofenac (Diclo; 10 mg/kg, i.p.), and xylazine were purchased from Daroopakhsh (Iran). Acetic acid and formalin were purchased from Merck (Germany). Cnicin isolated from Cnicus benedictus L., L-arg hydrochloride (L-arg HCL; 25, 50, 100 μg/paw), Nω-nitro-L-arg methyl ester hydrochloride (L-NAME; 25, 50, 100 μg/paw), sodium nitroprusside (SNP; 125, 250, 500 μg/paw), methylene blue (MB; 100, 200, 400 μg/paw), glibenclamide (Gli; 25, 50, 100 μg/paw), naltrindole (NAL; 0.99 mg/kg, i.p.), nor-binaltorphimine (NBT; 1.03 mg/kg, i.p.), naloxonazine (NAX; 3.5 mg/kg, i.p.), and glutamate all were purchased from Sigma Aldrich Company; USA.
Nor binaltorphimine
Manufactured by Merck Group
Sourced in United States
Nor-binaltorphimine is a laboratory chemical compound used in research and development applications. It is a potent and selective kappa opioid receptor antagonist. The core function of Nor-binaltorphimine is to serve as a research tool for studying the role of kappa opioid receptors in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Naltrindole, by Merck Group (5 mentions)
Glibenclamide, by Merck Group (2 mentions)
Naloxone, by Merck Group (2 mentions)
β funaltrexamine, by Merck Group (2 mentions)
Glutamate, by Merck Group (1 mentions)
Methylene blue, by Merck Group (1 mentions)
Sodium nitroprusside, by Merck Group (1 mentions)
Acetic acid, by Merck Group (1 mentions)
Formalin, by Merck Group (1 mentions)
Bestatin, by Bio-Techne (1 mentions)
Naltrindole, by Bio-Techne (1 mentions)
Naloxone, by Bio-Techne (1 mentions)
Am251, by Bio-Techne (1 mentions)
Am630, by Bio-Techne (1 mentions)
Nor binaltorphimine, by Bio-Techne (1 mentions)
Clocinnamox, by Bio-Techne (1 mentions)
Am251, by Merck Group (1 mentions)
Bestatin, by Merck Group (1 mentions)
Morphine sulfate, by Merck Group (1 mentions)
Hexamethonium, by Merck Group (1 mentions)
9 protocols using nor binaltorphimine
1
Antinociceptive Effects of CBHE and Cnicin
2
Intracerebroventricular Drug Administration in Rodents
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The following drugs and chemicals were used: PnPP-19 (synthesized by China Peptides, China), the opioid receptor antagonist naloxone (Sigma, USA), the μ-opioid receptor antagonist clocinnamox (Tocris, USA), the δ-opioid receptor antagonist naltrindole (Tocris, USA), the κ-opioid receptor antagonist nor-binaltorphimine (Sigma, USA), the aminopeptidase inhibitor bestatin (Sigma, USA), AM251 [N-(piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide; Tocris, USA], AM630 {6-iodo-2-methyl-1-[2-(4-morpholinyl)ethyl]-1H-indol-3-yl(4-ethoxyphenyl) methanone; Tocris, USA}, MAFP [(5Z,8Z,11Z,14Z)-5,8,11,14-eicosatetraenyl-methyl ester phosphonofluoridic acid, Tocris, USA] and VDM11 [(5Z,8Z,11Z,14Z)-N-(4-Hydroxy-2-methylphenyl)-5,8,11,14-eicosatetraenamide, Tocris, USA].
The drugs were dissolved as follows: PnPP-19 (saline), naloxone (saline), clocinnamox (saline), naltrindole (saline), nor-binaltorphimine (saline), bestatin (saline), AM251 and AM630 (12% DMSO in saline), MAFP (10% DMSO in saline), VDM11 (10% in saline) and injected at a volume of 2 μL into the lateral ventricle. The saline used for dilution of all drugs contained 0.5% Evans Blue.
The drugs were dissolved as follows: PnPP-19 (saline), naloxone (saline), clocinnamox (saline), naltrindole (saline), nor-binaltorphimine (saline), bestatin (saline), AM251 and AM630 (12% DMSO in saline), MAFP (10% DMSO in saline), VDM11 (10% in saline) and injected at a volume of 2 μL into the lateral ventricle. The saline used for dilution of all drugs contained 0.5% Evans Blue.
3
Solvent Effects on TPI Compound Assays
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In all assays, TPI compounds were dissolved
in 10% dimethyl sulfoxide, a concentration that did not produce any
detectable behavioral effect. Morphine sulfate, naloxone, nor-binaltorphimine
(nor-BNI), and naltrindole were purchased from Sigma-Aldrich (St.
Louis, MO) and dissolved in 0.9% sterile saline.
in 10% dimethyl sulfoxide, a concentration that did not produce any
detectable behavioral effect. Morphine sulfate, naloxone, nor-binaltorphimine
(nor-BNI), and naltrindole were purchased from Sigma-Aldrich (St.
Louis, MO) and dissolved in 0.9% sterile saline.
4
Anesthetic Agents and Sodium Chloride
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The drugs used in this study, alpha-chloralose, urethane, inactin, U50, norbinaltorphimine, and hexamethonium, were all purchased from Sigma Aldrich. Sodium chloride 0.9% was obtained from Fisher Scientific (Pittsburgh, PA). Isoflurane (ISOTHESIA) was purchased through the University of Texas from Henry Schein Animal Health (Dublin, OH).
5
Pharmacological Modulation of Kappa Opioid Receptor
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U50,488H (U50) (Sigma U111; MilliporeSigma, Saint Louis, MO), naltrexone hydrochloride (NTX) (Sigma N3136), norbinaltorphimine (norBNI) (Sigma N1771), naloxone methiodide (NLXM) (Sigma N129), and SR59230A0A hydrochloride (Bio-Techne 1511; Minneapolis, MN) were dissolved in 0.9% saline. Prior thermal imaging experiments, animals were habituated in a clear behavior chamber on a platform.34 (link) After habituation, animals were injected intraperitoneally (i.p.) with vehicle or antagonist, and 30 min later, vehicle or U50 (2, 5, or 10 mg/kg) was injected i.p. For experiments using intracerebroventricular administration of compounds, animals were similarly habituated. The cannula dummy was unscrewed from the guide cannula and the internal injector cannula was placed inside the guide cannula and screwed on securely. Over the course of 5 min, 5 μL of vehicle or 1 nmol norBNI were infused into the ventricle; after 30 min, 5 mg/kg U50 were administered i.p. NLXM was selected as a peripheral restricted antagonists due to reports that in mice methylnaltrexone is rapidly metabolized to naloxone and crosses the blood brain barrier.71 (link)
6
Pharmacological Agents for Cell Studies
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The drugs apamin, charybdotoxin, tetraethylammonium chloride, atropine, haloperidol, pindolol, yohimbine, prazosin, phenylpherine, clonidine, caffeine, glibenclamide, β-funaltrexamine, naltrindole, nor-binaltorphimine, all purchased from Sigma Aldrich (U.S.A.) and bradykinin (Tocris Bioscience, U.K.), were prepared at the desired dose by dissolving them in distilled water (dH2O). Phorbol 12-myristate 13-acetate (Sigma Aldrich, USA) was dissolved in PBS solution. Acetic acid and dimethyl sulfoxide (DMSO) were purchased from Fisher Scientific (U.K.).
7
Combinatorial Cytotoxicity Assay
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OVCAR4 and Ov4Carbo cells (1000/well in triplicate) were seeded in 96-well plates (triplicate) and treated with chloroxine (10 μM) ± carboplatin as before (see synergy validation, n = 8 biological repeats). The selective κ-opioid agonist: U50,488 ((±)-trans-U-50488 methanesulfonate salt, Sigma-Aldrich) and antagonist: nor-binaltorphimine (nor-BNI, Sigma-Aldrich) were used at 10 μM in place of chloroxine and in combination with carboplatin on day 5. Control plates displaying dose–response curves for each individual compound were seeded in parallel. Cell viability was determined on day 7 using CellTiter-Glo assay and dose–response curves generated.
8
Pharmacological Intervention in Pain Study
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Tetraethylammonium, glibenclamide, apamin, and charybdotoxin were purchased from Tocris (Bristol, UK). DMSO and Tween 20 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Naloxone hydrochloride, β-funaltrexamine, naltrindole, nor-binaltorphimine, DMSO, and Tween 20 were bought from Sigma-Aldrich (St. Louis, MO, USA). Morphine sulphate was purchased from Lipomed (Cambridge, MA, USA). All drugs were dissolved in 0.9% NaCl. The vehicle consisted of DMSO, Tween 20, and 0.9% NaCl at a ratio of 5:5:90. All treatments were administered either intraperitoneally or subcutaneously at a volume of 10 mL/kg of body weight. Intraperitoneal injection was administered in the intraperitoneal cavity and subcutaneous administrations were made into the loose skin over the interscapular area.
9
Synthesis and Pharmacological Evaluation
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14-O-methylmorphine (Fig. 1) was synthesized as described under Section 2.3. Tris-HCl, EGTA, NaCl, MgCl 2 × 6H 2 O, GDP, the GTP analogue GTPγS, the DOR and KOR antagonist naltrindole and norbinaltorphimine, respectively and the KOR agonist U-69593 were purchased from Sigma-Aldrich (Budapest, Hungary). The MOR selective antagonist cyprodime was provided by Dr.
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