Cb 13

Manufactured by Cayman Chemical

CB-13 is a chemical reagent used for research purposes. It functions as an inhibitor, with its core role being the modulation of specific biological processes. Detailed information about its intended applications is not provided to maintain an unbiased and factual approach.

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4 protocols using cb 13

Mitochondrial Function Assay Protocol

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Endothelin-1 (ET1), compound C, β-actin antibody, l-carnitine hydrochloride, oligomycin, carbonylcyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP), rotenone, antimycin A, and etomoxir were from Sigma Aldrich (St Louis, MO). CB-13 and the JC-1 mitochondrial membrane potential assay kit were from Cayman Chemical (Ann Arbor, MI). Calcein-AM (Molecular Probes) and carnitine palmitoyltransferase (CPT)-1β primers were from Life Technologies (Carlsbad, CA). p-AMPKα (Thr172) and AMPKα antibodies were from Cell Signaling (2535S and 2603S, respectively; Whitby, Canada). CB1 and CB2 antibodies were from Abcam (ab23703 and ab45942, respectively; Toronto, Canada). Proliferator-activated receptor-gamma coactivator (PGC)-1α antibody was from EMD Millipore (ST1202; Temecula, CA). XF24 FluxPaks were from Agilent (Santa Clara, CA).

Lu Y., Lee D.I., Roy Chowdhury S., Lu P., Kamboj A., Anderson C.M., Fernyhough P, & Anderson H.D. (2019). Activation of Cannabinoid Receptors Attenuates Endothelin-1–Induced Mitochondrial Dysfunction in Rat Ventricular Myocytes. Journal of Cardiovascular Pharmacology, 75(1), 54-63.

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Characterization of Mitochondrial Function

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AngII was from Sigma Aldrich (St. Louis, MO). CB13 (1-Naphthalenyl[4-(pentyloxy)-1-naphthalenyl]methanone), compound C (dosmorphin, (6-[4-(2-Piperidin-1-ylethoxy) phenyl]-3-pyridin-4-ylpyrazolo [1,5-a]pyrimidine)), JC-1 (tetraethylbenzimidazolylcarbocyanine iodide) Mitochondrial Membrane Potential assay kit and Mitochondrial Permeability Transition Pore (mPTP) assay kit were from Cayman Chemical (Ann Arbor, MI). Primary antibodies from Cell Signaling Technology (Danvers, Massachusetts) for phospho-AMPKα (Thr172) (1:1000, cat. 2535), AMPKα (1:1000, cat. 2603), phospho-LKB1 (1:1000, cat. 3482), LKB1 (1:1000, cat. 3047) and Abcam (Toronto, Ontario) connexin 43 (1:7500, cat. ab11370), CB1R (1:500, cat. ab23703), and CB2R (1:5000, cat. ab45942) were used.

Altieri D.I., Etzion Y, & Anderson H.D. (2023). Cannabinoid receptor agonist attenuates angiotensin II–induced enlargement and mitochondrial dysfunction in rat atrial cardiomyocytes. Frontiers in Pharmacology, 14, 1142583.

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CB13 Treatment in Cardiomyocytes

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CB13 (1-naphthalenyl [4-(pentyloxy)-1-naphthalenyl]methanone) was from Cayman Chemical (Ann Arbor, Michigan). Tyrode’s buffer was supplemented with vehicle (0.1% v/v DMSO) or CB13 (1 μM, 0.1% v/v DMSO). Treatments remained in buffer for the remainder of the experiment. The concentration of CB13 was based on our previous findings in cardiomyocytes in vitro (Lu et al., 2014 (link); Lu et al., 2020 (link)).

Lee D.I., Murninkas M., Elyagon S., Etzion Y, & Anderson H.D. (2021). Cannabinoid Receptor Agonist Inhibits Atrial Electrical Remodeling in a Tachypaced Ex Vivo Rat Model. Frontiers in Pharmacology, 12, 642398.

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Preparation of Cannabinoid Compounds for Behavioral and Physiological Studies

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Complete Freund’s adjuvant (CFA) (Thermo Fisher, St. Louis, MO) was dissolved in a 1:1 ratio of saline:CFA prior to intraplantar (i.pl) injection. CB-13, AM6545, AM630 and rimonabant (all from Cayman Chemical Company, Ann Arbor, MI) were dissolved in vehicle consisting of 20% DMSO (Sigma Aldrich, St. Louis, MO), 8% ethanol, 8% Tween 80 (Thermo Fisher, St. Louis, MO) and 64% saline and administered via intraperitoneal (i.p.) injection in a volume of 5 mL/kg for behavioral studies. CB-13, AM630 and rimonabant were dissolved in DMSO (25 mg/mL) to create stocks that were frozen until use. On the day of the experiment, stocks were diluted with a volume of DMSO to achieve a concentration of 20% DMSO in the final solution. 95% ethanol was then added, the solution vortexed again, followed by Tween 80 and finally saline. For AM6545, the steps were the same as the other compounds used, but the solution was sonicated for approximately 45 minutes to get the compound to fully dissolve (when the solution was clear). For calcium imaging and electrophysiology studies, prostaglandin E2 (PGE2) (Thermo Fisher, St. Louis, MO) and CB-13 were dissolved in DMSO and diluted in external recording solution.

Slivicki R.A., Yi J., Brings V.E., Huynh P.N, & Gereau RW I.V. (2021). The cannabinoid agonist CB-13 produces peripherally mediated analgesia in mice but elicits tolerance and signs of CNS activity with repeated dosing. Pain, 163(8), 1603-1621.

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