N6 cyclopentyladenosine cpa

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N6-cyclopentyladenosine (CPA) is a chemical compound that is commonly used as a selective agonist for the adenosine A1 receptor. It is a synthetic derivative of the natural nucleoside adenosine. CPA is often utilized in research applications to study the physiological effects and mechanisms of adenosine receptor activation.

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7 protocols using n6 cyclopentyladenosine cpa

Radioligand Binding Assay Protocols

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[³H]-DPCPX (8-Cyclopentyl-1,3-dipropylxanthine) (118 Ci/mmol) and [³H]-ZM 241385 ([2-³H](4-(2-[7-amino-2-(2-furyl) [1,2,4] triazolo [2,3-α] [1,3,5] triazin-5-ylamino] ethyl) phenol) 27.4 Ci/mmol) were purchased from Amersham Biosciences (Buckinghamshire, UK). Theophylline and hydrogen peroxide were acquired from Sigma Aldrich (Madrid, Spain). CPA (N⁶-Cyclopentyladenosine) was obtained from Tocris (Madrid, Spain). The Liquid scintillation cocktail was supplied by Perkin Elmer (Boston, MA, USA). Additional reagents were of analytical grade.

Alonso-Andrés P., Martín M, & Albasanz J.L. (2019). Modulation of Adenosine Receptors and Antioxidative Effect of Beer Extracts in in Vitro Models. Nutrients, 11(6), 1258.

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Adenosine Receptor Agonists and Methamphetamine

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The selective adenosine A₁ receptor agonist, CPA (N⁶-Cyclopentyladenosine), and selective adenosine A_2A receptor agonist CGS 21680 hydrochloride, (4-[2-[[6-Amino-9-(N-ethyl-β-D-ribofuranuronamidosyl)-9H-purin-2-yl]amino]ethyl]benzenepropanoic acid hydrochloride), were purchased from Tocris Bioscience (Bristol, UK). Methamphetamine hydrochloride was obtained from Sigma-Aldrich (St Louis, MO). All drugs were dissolved in sterile-filtered physiological saline.

Kavanagh K.A., Schreiner D.C., Levis S.C., O’Neill C.E, & Bachtell R.K. (2014). Role of adenosine receptor subtypes in methamphetamine reward and reinforcement. Neuropharmacology, 89, 265-273.

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Pharmacological Modulation of Adenosine Receptors

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All reagents were purchased from Fisher Scientific (Fair Lawn, NJ, USA) unless otherwise stated. Phosphate buffered saline (PBS) was used to calibrate electrodes containing (in mM): 131.25 NaCl, 3.0 KCl, 10.0 NaH₂PO₄, 1.2 MgCl₂, 2.0 Na₂SO₄, and 1.2 CaCl₂ with the pH adjusted to 7.4. Sodium phosphate was purchased from RICCA Chemical Company (Arlington, TX, USA). All aqueous solutions were prepared with deionized water (Milli-Q Biocel; Millipore, Billerica, MA, USA). Adenosine was prepared as a 10 mM stock solution in 0.1 M HClO₄ and stored in the refrigerator.
DPCPX (6 mg/kg, i.p., 8-cyclopentyl-1,3-dipropylxanthine, Sigma Aldrich) was dissolved in dimethylsulfoxide (DMSO). CPA (N⁶-cyclopentyladenosine) was purchased from Tocris Bioscience (Ellisville, MO, USA), dissolved in saline, and administered at 1 mg/kg. These doses were chosen as large doses that were previously used in the literature [12] (link), [16] (link), [17] (link).

Nguyen M.D., Lee S.T., Ross A.E., Ryals M., Choudhry V.I, & Venton B.J. (2014). Characterization of Spontaneous, Transient Adenosine Release in the Caudate-Putamen and Prefrontal Cortex. PLoS ONE, 9(1), e87165.

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Cellular Model for TGF-β Signaling

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EC cell lines, HEC-1-A (American Type Culture Collection) and HEC-50 (gift from Bryan Hennessy, University of Texas MD Anderson Cancer Center) were maintained as described (21 (link)). Recombinant human TGF-β1 (R&D Systems) was reconstituted in 4 mM hydrochloric acid (HCl) containing 0.1% bovine serum albumin (BSA). 5’AMP and adenosine 5′-(α,β-methylene) diphosphate (AoPCP) were purchased from Sigma-Aldrich and 5’-N-ethylcarboxamidoadenosine (NECA) and N6-cyclopentyladenosine (CPA) from Tocris Bioscience. Non-targeting and CD73 siRNA (1247) were purchased from Sigma-Aldrich (21 (link)). Human CCND1 esiRNA was purchased from Sigma-Aldrich.

Kurnit K.C., Draisey A., Kazen R.C., Chung C., Phan L.H., Harvey J.B., Feng J., Xie S., Broaddus R.R, & Bowser J.L. (2021). Loss of CD73 shifts transforming growth factor-β1 (TGF-β1) from tumor suppressor to promoter in endometrial cancer. Cancer letters, 505, 75-86.

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Pharmacological Modulation of Adenosine Signaling

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(R)-(+)-[2,3-Dihydro-5-methyl-3-(4-morpholinylmethyl)pyrrolo[1,2,3-de]-1,4-benzoxazin-6-yl]-1-naphthalenylmethanone mesylate (WIN55212-2), 8-cyclopentyl-1,3-dipropylxanthine (DPCPX), N6-cyclopentyladenosine (CPA), N-(piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide (AM251), rolipram, and forskolin were purchased from Tocris (Bristol, UK). Adenosine deaminase was obtained from Sigma-Aldrich. Stock solutions of WIN55212-2 (20 mM), rolipram (20 mM), DPCPX (50 μM), and AM251 (5 mM) were prepared in dimethyl sulfoxide (DMSO). forskolin (50 mM) and CPA (2 mM) stock solutions were prepared in ethanol and water, respectively. Suitable dilutions of each stock solution with Krebs buffer were made before performing the experiments.

Serpa A., Correia S., Ribeiro J.A., Sebastião A.M, & Cascalheira J.F. (2015). The Combined Inhibitory Effect of the Adenosine A1 and Cannabinoid CB1 Receptors on cAMP Accumulation in the Hippocampus Is Additive and Independent of A1 Receptor Desensitization. BioMed Research International, 2015, 872684.

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Adenosine Receptor Agonists in Research

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The following reagents were used in this study: adenosine, adenosine 5′-monophosphate (AMP), ovalbumin (grade V), urethane (all Sigma-Aldrich, UK), propofol, isoflurane (Centaur Services, UK) and the selective adenosine A₁ receptor agonist N⁶-cyclopentyladenosine (CPA) (Tocris, UK).

Lexmond A.J., Keir S., Terakosolphan W., Page C.P, & Forbes B. (2018). A novel method for studying airway hyperresponsiveness in allergic guinea pigs in vivo using the PreciseInhale system for delivery of dry powder aerosols. Drug Delivery and Translational Research, 8(3), 760-769.

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Synthesis and Characterization of Pannexin-based Compounds

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¹⁰Panx (WRQAAFVDSY) and ^scPanx (FSVYWAQADR) were synthesised by Thermo Fisher Scientific (purity, >95%); pyridoxalphosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS), N6-Cyclopentyladenosine (CPA), ZD7288, SCH58261, NBQX, and CPP were purchased from Tocris, Reactive Blue-2 (RB-2), Tolbutamide, SCH23390, 8-Cyclopentyl-1,3-dimethylxanthine (8-CPT), picrotoxin from Sigma-Aldrich and EtBr from Molecular Probes.

Vasile F., Dossi E., Moulard J., Ezan P., Lecoin L., Cohen-Salmon M., Mailly P., Le Bert M., Couillin I., Bemelmans A, & Rouach N. (2022). Pannexin 1 activity in astroglia sets hippocampal neuronal network patterns. PLOS Biology, 20(12), e3001891.

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