NECA (5'-N-Ethylcarboxamidoadenosine) is a novel and potent adenosine receptor agonist with Ki values of 6.2, 14, and 20 nM for human A3, A1 and A2A receptors, respectively, and an EC50 of 2.4 μM for human A2B. It can prevent platelet aggregation and is centrally active in vivo.
Physicochemical Properties
| Molecular Formula | C12H16N6O4 |
| Molecular Weight | 308.29324 |
| Exact Mass | 308.123 |
| CAS # | 35920-39-9 |
| PubChem CID | 448222 |
| Appearance | White to off-white solid powder |
| Density | 1.9±0.1 g/cm3 |
| Melting Point | 229-231ºC |
| Index of Refraction | 1.829 |
| LogP | -0.48 |
| Hydrogen Bond Donor Count | 4 |
| Hydrogen Bond Acceptor Count | 8 |
| Rotatable Bond Count | 3 |
| Heavy Atom Count | 22 |
| Complexity | 426 |
| Defined Atom Stereocenter Count | 4 |
| SMILES | O[C@H]1[C@H](N2C=NC3=C(N)N=CN=C23)O[C@H](C(NCC)=O)[C@H]1O |
| InChi Key | JADDQZYHOWSFJD-FLNNQWSLSA-N |
| InChi Code | InChI=1S/C12H16N6O4/c1-2-14-11(21)8-6(19)7(20)12(22-8)18-4-17-5-9(13)15-3-16-10(5)18/h3-4,6-8,12,19-20H,2H2,1H3,(H,14,21)(H2,13,15,16)/t6-,7+,8-,12+/m0/s1 |
| Chemical Name | (2S,3S,4R,5R)-5-(6-aminopurin-9-yl)-N-ethyl-3,4-dihydroxyoxolane-2-carboxamide |
| Synonyms | 5'-N-Ethylcarboxamidoadenosine; NECA |
| HS Tariff Code | 2934.99.9001 |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets | Adenosine receptor; IL-18; TNF-α; CD54; JNK |
| ln Vitro | 5'-N-Ethylcarboxamidoadenosine (NECA) administration results in a dose-dependent significant decrease in the mean number of cocaine infusions obtained per session [5'-N-Ethylcarboxamidoadenosine (NECA): F(4,12)=14.9; P<0.001]. Significantly higher latencies than those obtained for vehicle treatment are observed upon administration of 5'-N-Ethylcarboxamidoadenosine (NECA) [F(4,12)=16.1; P<0.001][1]. In diabetic rats, daily intraperitoneal injections of 5'-N-Ethylcarboxamidoadenosine (NECA) at 0.3 mg/kg/day for two weeks lower MDA levels; control rats do not experience this same reduction. In diabetic rats, daily administration of NECA (0.3 mg/kg/day, i.p.) for two weeks decreases the gene expression of TNF-α and interleukin (IL)-18 induced by diabetes, while having no effect on control rats. For two weeks, a daily intraperitoneal injection of 5'-N-Ethylcarboxamidoadenosine (NECA) at a dose of 0.3 mg/kg/day inhibits the activation of JNK MAPK in diabetic rats, while having no effect on control rats[2]. |
| ln Vivo | The mean number of cocaine infusions per injection was significantly reduced in a dose-dependent manner after administration of 5'-N-ethylcarboxamideadenosine (NECA) [5'-N-ethylcarboxamideadenosine (NECA): F(4,12)= 14.9; P<0.001]. Administration of 5'-N-ethylcarboxamide adenosine (NECA) [F(4,12)=16.1; P<0.001] resulted in a significant increase in latency above the values obtained with vehicle treatment [1]. Daily intraperitoneal injection of 5'-N-ethylcarboxamide adenosine (NECA) at a dose of 0.3 mg/kg/day for two weeks reduced malondialdehyde (MDA) levels in diabetic rats but did not affect controls rat. Daily treatment with NECA (0.3 mg/kg/day, intraperitoneally for two weeks) reduced diabetes-induced gene expression of tumor necrosis factor (TNF)-α and interleukin (IL)-18 in diabetic rats, but not Affected control rats. Daily intraperitoneal injection of 5'-N-ethylcarboxamide adenosine (NECA) 0.3 mg/kg/day for two weeks also blocked the activation of JNK MAPK in diabetic rats but did not affect control rats [2 ]. |
| Animal Protocol | For this experiment, male Wistar rats are utilized. The weight of these animals varies from 350 to 425 g. They live in separate hanging wire cages with a 12-hour light/dark cycle. Four rats are used to test the effects of 5'-N-Ethylcarboxamidoadenosine (NECA). 15 minutes before the start of the test sessions, either 5'-N-Ethylcarboxamidoadenosine (NECA) (5, 7.5, 10, 20 μg/kg) or vehicle (saline) is given intraperitoneally. A random order crossover design is used in the administration of 5'-N-Ethylcarboxamidoadenosine (NECA). Animals are tested twice with the same dosage in the majority of cases[1]. |
| References |
[1]. Adenosine agonists CGS 21680 and NECA inhibit the initiation of cocaine self-administration. Pharmacol Biochem Behav. 2001 Apr;68(4):797-803. [2]. Reno-protective effect of NECA in diabetic nephropathy: implication of IL-18 and ICAM-1. Eur Cytokine Netw. 2012 Jul-Sep;23(3):78-86. |
| Additional Infomation |
N-ethyl-5'-carboxamidoadenosine is a derivative of adenosine in which the 5'-hydroxymethyl group is replaced by an N-ethylcarboxamido group. It has a role as an adenosine A1 receptor agonist, an adenosine A2A receptor agonist, an EC 3.1.4.* (phosphoric diester hydrolase) inhibitor, an antineoplastic agent and a vasodilator agent. It is a member of adenosines and a monocarboxylic acid amide. It is functionally related to an adenosine. A stable adenosine A1 and A2 receptor agonist. Experimentally, it inhibits cAMP and cGMP phosphodiesterase activity. |
Solubility Data
| Solubility (In Vitro) | DMSO: 62~62.5 mg/mL (201.1~202.7 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (8.11 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (8.11 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 3: ≥ 2.5 mg/mL (8.11 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.2437 mL | 16.2185 mL | 32.4370 mL | |
| 5 mM | 0.6487 mL | 3.2437 mL | 6.4874 mL | |
| 10 mM | 0.3244 mL | 1.6218 mL | 3.2437 mL |