5'-N-Ethylcarboxamidoadenosine
Based on 8 publication(s) in Google Scholar
5'-N-Ethylcarboxamidoadenosine (NECA) is a nonselective adenosine receptor agonist.
For research use only. We do not sell to patients.
- Purity: 99.09%
- CAS No.: 35920-39-9
- Formula: C12H16N6O4
- Molecular Weight:308.29
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Storage:Powder -20°C, 3 years , 4°C, 2 years ; In solvent -80°C, 2 years , -20°C, 1 year
Publications Citing Use of MedChemExpress (MCE) 5'-N-Ethylcarboxamidoadenosine
More- Nat Commun. 2026 Mar 31;17(1):3045. [Abstract]
- Cell Death Dis. 2020 Mar 23;11(3):202. [Abstract]
- J Med Chem. 2024 Mar 28;67(6):5075-5092. [Abstract]
- J Med Chem. 2023 Apr 13;66(7):4734-4754. [Abstract]
- J Med Chem. 2022 Mar 10;65(5):4367-4386. [Abstract]
- Pharmaceutics. 2021 Jul 27;13(8):1152. [Abstract]
- Nanomaterials. 2021 Apr 24;11(5):1102. [Abstract]
- bioRxiv. 2026 Mar 30.
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WB
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RT-PCR
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Cell Imaging/Staining
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Cell Imaging/Staining
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RT-PCR
All Adenosine Receptor Isoforms
More
Biological Activity
Adenosine receptor[1]
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Cell Line
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Type | Value | Description | References |
|---|---|---|---|---|
| CHO | EC50 |
2.4 μM
Compound: NECA
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Receptor stimulated adenylyl cyclase activity in CHO cells expressing human recombinant Adenosine A2B receptor
Receptor stimulated adenylyl cyclase activity in CHO cells expressing human recombinant Adenosine A2B receptor
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[PMID: 11459663] |
| CHO | EC50 |
0.04 μM
Compound: NECA
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G-protein activation in human Adenosine A2A receptor expressing CHO cells using cAMP assay
G-protein activation in human Adenosine A2A receptor expressing CHO cells using cAMP assay
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[PMID: 11784146] |
| CHO | EC50 |
84.8 nM
Compound: NECA
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Effective concentration for stimulation of [35S]GTP-gamma-S, binding at human adenosine A3 receptor expressed in CHO cells
Effective concentration for stimulation of [35S]GTP-gamma-S, binding at human adenosine A3 receptor expressed in CHO cells
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[PMID: 15214785] |
| CHO | EC50 |
104 nM
Compound: NECA
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Agonistic activity against human adenosine A2B receptor expressed in CHO cells
Agonistic activity against human adenosine A2B receptor expressed in CHO cells
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[PMID: 15239649] |
| CHO | EC50 |
440 nM
Compound: NECA
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Potency against cAMP formation in CHO cells expressing recombinant human A2B receptor
Potency against cAMP formation in CHO cells expressing recombinant human A2B receptor
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[PMID: 15267242] |
| CHO | EC50 |
50 nM
Compound: NECA
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Potency against cAMP formation in CHO cells expressing recombinant human A2A receptor
Potency against cAMP formation in CHO cells expressing recombinant human A2A receptor
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[PMID: 15267242] |
| CHO | EC50 |
160 nM
Compound: 1, NECA
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Activity at human adenosine A2B receptor expressed in CHO cells assessed as stimulation of cAMP levels
Activity at human adenosine A2B receptor expressed in CHO cells assessed as stimulation of cAMP levels
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[PMID: 17228880] |
| CHO | EC50 |
155 nM
Compound: NECA
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Agonist activity at human adenosine A2B receptor expressed in CHO cells assessed as stimulation of cAMP production
Agonist activity at human adenosine A2B receptor expressed in CHO cells assessed as stimulation of cAMP production
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[PMID: 17306548] |
| CHO | EC50 |
21.9 nM
Compound: 2, NECA
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Agonist activity at human adenosine A2A receptor expressed in CHO cells assessed as stimulation of cAMP production
Agonist activity at human adenosine A2A receptor expressed in CHO cells assessed as stimulation of cAMP production
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[PMID: 17378544] |
| CHO | EC50 |
30 nM
Compound: NECA
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Agonist activity at human adenosine A1 receptor expressed in CHO cells by GTPPgammaS binding assay
Agonist activity at human adenosine A1 receptor expressed in CHO cells by GTPPgammaS binding assay
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[PMID: 20031406] |
| CHO | EC50 |
38 nM
Compound: NECA
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Agonist activity at human adenosine A2B receptor expressed in CHO cells assessed as effect on cAMP production by luciferase reporter gene assay
Agonist activity at human adenosine A2B receptor expressed in CHO cells assessed as effect on cAMP production by luciferase reporter gene assay
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[PMID: 20031406] |
| CHO | EC50 |
63 nM
Compound: NECA
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Agonist activity at human adenosine A3 receptor expressed in CHO cells by GTPPgammaS binding assay
Agonist activity at human adenosine A3 receptor expressed in CHO cells by GTPPgammaS binding assay
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[PMID: 20031406] |
| CHO | EC50 |
10.2 nM
Compound: NECA
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Agonist activity at human recombinant adenosine receptor-1 expressed in CHO cells assessed as inhibition of forskolin-stimulated cAMP production preincubated for 45 mins prior forskolin-induction measured after 15 mins by immunoassay
Agonist activity at human recombinant adenosine receptor-1 expressed in CHO cells assessed as inhibition of forskolin-stimulated cAMP production preincubated for 45 mins prior forskolin-induction measured after 15 mins by immunoassay
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[PMID: 21858244] |
| CHO | EC50 |
0.018 μM
Compound: 2 (NECA)
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Agonist activity at human recombinant adenosine A2A receptor expressed in CHO cells assessed as [3H]cAMP accumulation after 15 mins by liquid scintillation counting
Agonist activity at human recombinant adenosine A2A receptor expressed in CHO cells assessed as [3H]cAMP accumulation after 15 mins by liquid scintillation counting
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[PMID: 23245803] |
| CHO | EC50 |
12.58 nM
Compound: 1b, NECA
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Agonist activity at human adenosine A2A receptor expressed in CHO cells assessed as stimulation of [3H]cAMP levels by scintillation counting method
Agonist activity at human adenosine A2A receptor expressed in CHO cells assessed as stimulation of [3H]cAMP levels by scintillation counting method
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[PMID: 25780876] |
| CHO | EC50 |
>10000 nM
Compound: 1b, NECA
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Agonist activity at human adenosine A2B receptor expressed in CHO cells assessed as stimulation of [3H]cAMP levels by scintillation counting method
Agonist activity at human adenosine A2B receptor expressed in CHO cells assessed as stimulation of [3H]cAMP levels by scintillation counting method
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[PMID: 25780876] |
| CHO | EC50 |
45 nM
Compound: NECA
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Agonist activity at human recombinant adenosine A2A receptor expressed in CHO cells assessed as stimulation of NECA-mediated cAMP accumulation preincubated for 15 mins followed by NECA addition measured after 15 mins by enzyme immunoassay
Agonist activity at human recombinant adenosine A2A receptor expressed in CHO cells assessed as stimulation of NECA-mediated cAMP accumulation preincubated for 15 mins followed by NECA addition measured after 15 mins by enzyme immunoassay
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10.1039/C5MD00086F |
| CHO-K1 | EC50 |
17.6 nM
Compound: 2, NECA
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Agonist activity at human adenosine A2A receptor expressed in CHO-K1 cells by cAMP accumulation assay
Agonist activity at human adenosine A2A receptor expressed in CHO-K1 cells by cAMP accumulation assay
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[PMID: 24900277] |
| COS-7 | EC50 |
0.078 μM
Compound: NECA
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Activity against human wild type adenosine A3 receptor expressed in COS7 cells as measured by accumulation of inositol phosphate by PLC assay
Activity against human wild type adenosine A3 receptor expressed in COS7 cells as measured by accumulation of inositol phosphate by PLC assay
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[PMID: 16640329] |
| COS-7 | EC50 |
10.4 μM
Compound: NECA
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Activity against human adenosine A3 receptor H272E mutant expressed in COS7 cells as measured by accumulation of inositol phosphate by PLC assay
Activity against human adenosine A3 receptor H272E mutant expressed in COS7 cells as measured by accumulation of inositol phosphate by PLC assay
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[PMID: 16640329] |
| HCT-116 | IC50 |
>10 μM
Compound: NECA
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Cytotoxicity against human HCT-116 cells assessed as cell viability incubated for 72 to 96 hrs by MTT assay
Cytotoxicity against human HCT-116 cells assessed as cell viability incubated for 72 to 96 hrs by MTT assay
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[PMID: 35707146] |
| HEK293 | IC50 |
3.7 x 10-8 M
Compound: NECA
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Displacement of [3H]CGS 21680 from human recombinant adenosine A2a receptor expressed in HEK293 cells
Displacement of [3H]CGS 21680 from human recombinant adenosine A2a receptor expressed in HEK293 cells
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[PMID: 26988801] |
| HEK293 | IC50 |
3.7 x 10-8 M
Compound: NECA
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Displacement of [3H]CGS 21680 from human recombinant adenosine receptor A2A expressed in HEK293 cells measured after 120 mins by scintillation counting method
Displacement of [3H]CGS 21680 from human recombinant adenosine receptor A2A expressed in HEK293 cells measured after 120 mins by scintillation counting method
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[PMID: 27876250] |
| HEK293 | IC50 |
3.7 x 10-2 μM
Compound: NECA
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Displacement of [3H]CGS 21680 from human recombinant adenosine receptor A2A expressed in HEK293 cells measured after 120 mins by scintillation counting method
Displacement of [3H]CGS 21680 from human recombinant adenosine receptor A2A expressed in HEK293 cells measured after 120 mins by scintillation counting method
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[PMID: 27876250] |
| HEK293 | EC50 |
188 nM
Compound: 37; NECA
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Partial agonist activity at recombinant mouse A3AR expressed in HEK293 cell membranes assessed as stimulation of [35S]GTPgammaS binding measured after 2 hrs in presence of A2BAR blocker PSB-603 by liquid scintillation counting analysis
Partial agonist activity at recombinant mouse A3AR expressed in HEK293 cell membranes assessed as stimulation of [35S]GTPgammaS binding measured after 2 hrs in presence of A2BAR blocker PSB-603 by liquid scintillation counting analysis
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[PMID: 32271569] |
| PBMC | IC50 |
187 nM
Compound: NECA
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Antiinflammatory activity in rat PBMC assessed as inhibition of LPS-induced TNFalpha production after 20 hrs by ELISA
Antiinflammatory activity in rat PBMC assessed as inhibition of LPS-induced TNFalpha production after 20 hrs by ELISA
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[PMID: 20031406] |
| PBMC | IC50 |
40 nM
Compound: NECA
|
Antiinflammatory activity in human PBMC assessed as inhibition of LPS-induced TNFalpha production after 20 hrs by ELISA
Antiinflammatory activity in human PBMC assessed as inhibition of LPS-induced TNFalpha production after 20 hrs by ELISA
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[PMID: 20031406] |
| PC-12 | EC50 |
130 nM
Compound: 1
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Potency against PC12 cell A2 adenosine receptor by adenylate cyclase activation
Potency against PC12 cell A2 adenosine receptor by adenylate cyclase activation
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[PMID: 3018244] |
| PC-3 | IC50 |
>10 μM
Compound: NECA
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Cytotoxicity against human PC-3 cells assessed as cell viability incubated for 72 to 96 hrs by MTT assay
Cytotoxicity against human PC-3 cells assessed as cell viability incubated for 72 to 96 hrs by MTT assay
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[PMID: 35707146] |
| WI-38 | IC50 |
>10 μM
Compound: NECA
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Cytotoxicity against human WI-38 cells assessed as cell viability incubated for 72 to 96 hrs by MTT assay
Cytotoxicity against human WI-38 cells assessed as cell viability incubated for 72 to 96 hrs by MTT assay
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[PMID: 35707146] |
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Chemical Information
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CAS No. 35920-39-9
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Appearance Solid
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Molecular Weight 308.29
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Formula C12H16N6O4
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Color White to off-white
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SMILES
O[C@H]1[C@H](N2C=NC3=C(N)N=CN=C23)O[C@H](C(NCC)=O)[C@H]1O
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Synonyms
NECA
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
Powder -20°C 3 years 4°C 2 years In solvent -80°C 2 years -20°C 1 year
Publications (8)
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Journal Impact Factor
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Most Recent
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Nat Commun
Timosaponin AIII enhances CAR-T cell potency and prevents relapse through impairing CAR-Tregs. [Abstract]2026 Mar 31;17(1):3045. PMID: 41916982 -
Cell Death Dis
CD73 promotes tumor metastasis by modulating RICS/RhoA signaling and EMT in gastric cancer. [Abstract]2020 Mar 23;11(3):202. PMID: 32205841
5'-N-Ethylcarboxamidoadenosine purchased from MedChemExpress. Usage Cited in: Cell Death Dis. 2020 Mar 23;11(3):202. [Abstract]
OT-I cytotoxic T lymphocytes and MC38-OVA were cocultured with 5'-N-Ethylcarboxamidoadenosine (NECA) (1 μM; 48 h). 5'-N-Ethylcarboxamidoadenosine (NECA), a potent immunosuppressive ligand, notably impaired the effect that OT-I CTLs killed MC38-OVA cancer cells.
5'-N-Ethylcarboxamidoadenosine purchased from MedChemExpress. Usage Cited in: Cell Death Dis. 2020 Mar 23;11(3):202. [Abstract]
Human Jurkat T cells were stimulated with 20 ng/mL PMA and 1 μg/mL ionomycin (IONO) in the presence of 5'-N-Ethylcarboxamidoadenosine (NECA) (1 μM) for 5 h with varied concentrations of compound 7ai (0.1, 1,10 μM). The gene expression of PD-1 and TIM-3 were determined by Q-PCR and standardized using GAPDH as an internal reference.
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J Med Chem
Subtle Structural Changes across the Boundary between A2AR/A2BR Dual Antagonism and A2BR Antagonism: A Novel Class of 2-Aminopyrimidine-Based Derivatives. [Abstract]2024 Mar 28;67(6):5075-5092. PMID: 38483150
5'-N-Ethylcarboxamidoadenosine purchased from MedChemExpress. Usage Cited in: J Med Chem. 2024 Mar 28;67(6):5075-5092. [Abstract]
WB analysis of inhibition effects of CD73 plasmids, 5'-N-Ethylcarboxamidoadenosine (NECA) (10 μM), or adenosine in RhoA/LIMK/cofilin signaling. 5'-N-Ethylcarboxamidoadenosine (NECA) inhibited the phosphorylation of RhoA-LIMK-cofilin signaling and showed the reduction of p-LIMK and p-cofilin to the maximum extent in AGS cells.
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J Med Chem
Discovery of Pyridinone Derivatives as Potent, Selective, and Orally Bioavailable Adenosine A2A Receptor Antagonists for Cancer Immunotherapy. [Abstract]2023 Apr 13;66(7):4734-4754. PMID: 36951370
5'-N-Ethylcarboxamidoadenosine purchased from MedChemExpress. Usage Cited in: J Med Chem. 2023 Apr 13;66(7):4734-4754. [Abstract]
5'-N-Ethylcarboxamidoadenosine (NECA) (1 μM) treatment significantly down-regulated the gene expressions of GZMB, IFNG, and IL-2 in Jurkat T cells.
5'-N-Ethylcarboxamidoadenosine purchased from MedChemExpress. Usage Cited in: J Med Chem. 2023 Apr 13;66(7):4734-4754. [Abstract]
Cytotoxic T lymphocytes from OT-I mouse splenocyte (OT-I CTL) were co-incubated with MC38-OVA cells for 48 h in the presence of 5'-N-Ethylcarboxamidoadenosine (NECA) (1 μM) with or without compound 38. Then, living MC38-OVA cells were stained with a cell-permeant dye Calcein AM with green fluorescence. 5'-N-Ethylcarboxamidoadenosine (NECA) profoundly impaired the cytotoxic function of OT-ICTL.
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J Med Chem
Design, Synthesis, and Bioevaluation of 2-Aminopteridin-7(8 H)-one Derivatives as Novel Potent Adenosine A2A Receptor Antagonists for Cancer Immunotherapy. [Abstract]2022 Mar 10;65(5):4367-4386. PMID: 35212542 -
Pharmaceutics
Effective Actions of Ion Release from Mesoporous Bioactive Glass and Macrophage Mediators on the Differentiation of Osteoprogenitor and Endothelial Progenitor Cells. [Abstract]2021 Jul 27;13(8):1152. PMID: 34452110 -
Nanomaterials
Effects of Ipriflavone-Loaded Mesoporous Nanospheres on the Differentiation of Endothelial Progenitor Cells and Their Modulation by Macrophages. [Abstract]2021 Apr 24;11(5):1102. PMID: 33923311 -
Solvent & Solubility
DMSO : 62.5 mg/mL (202.73 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
H2O : < 0.1 mg/mL (insoluble)
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
Select the appropriate dissolution method based on your experimental animal and administration route.
- For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
- To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
- The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
Solubility: ≥ 2.5 mg/mL (8.11 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)
Solubility: ≥ 2.5 mg/mL (8.11 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
Please enter the basic information of animal experiments:
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
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%DMSO +
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
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%+
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+%Tween-80 + +
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%Saline +
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Working solution concentration: 0.22 mg/mL
Method for preparing stock solution: mg drug dissolved in μL DMSO. Stock solution concentration: mg/mL.
1. Take μL DMSO stock solution;
2. Add μL .
μL , mix evenly;
3. Then add μL Tween 80, mix evenly;
4. Then add μL
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Protocol
Male Wistar rats are used in this experiment. These animals range in weight between 350 and 425 g. They are housed individually in hanging wire cages under a 12-h light/dark cycle. The effects of 5'-N-Ethylcarboxamidoadenosine (NECA) are tested in four rats. In the experiment, either 5'-N-Ethylcarboxamidoadenosine (NECA) (5, 7.5, 10, 20 μg/kg) or vehicle (saline) is administered intraperitoneally 15 min prior to the start of test sessions. 5'-N-Ethylcarboxamidoadenosine (NECA) is administered following a random order crossover design. In most cases, animals are tested twice with the same dose[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Purity & Documentation
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Data Sheet (277 KB)
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SDS (479 KB)
- English - EN (479 KB)
- Français - FR (479 KB)
- Deutsch - DE (479 KB)
- Norwegian - NO (479 KB)
- Español - ES (479 KB)
- Swedish - SV (479 KB)
- Italian - IT (479 KB)
- Korean - KR (479 KB)
- Portuguese - PT (479 KB)
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Handling Instructions (2659 KB)
References
[1]. Knapp CM, et al. Adenosine agonists CGS 21680 and NECA inhibit the initiation of cocaine self-administration. Pharmacol Biochem Behav. 2001 Apr;68(4):797-803. [Content Brief]
[2]. Elsherbiny NM, et al. 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. [Content Brief]
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| DMSO | 1 mM | 3.2437 mL | 16.2185 mL | 32.4370 mL | 81.0925 mL |
| 5 mM | 0.6487 mL | 3.2437 mL | 6.4874 mL | 16.2185 mL | |
| 10 mM | 0.3244 mL | 1.6218 mL | 3.2437 mL | 8.1092 mL | |
| 15 mM | 0.2162 mL | 1.0812 mL | 2.1625 mL | 5.4062 mL | |
| 20 mM | 0.1622 mL | 0.8109 mL | 1.6218 mL | 4.0546 mL | |
| 25 mM | 0.1297 mL | 0.6487 mL | 1.2975 mL | 3.2437 mL | |
| 30 mM | 0.1081 mL | 0.5406 mL | 1.0812 mL | 2.7031 mL | |
| 40 mM | 0.0811 mL | 0.4055 mL | 0.8109 mL | 2.0273 mL | |
| 50 mM | 0.0649 mL | 0.3244 mL | 0.6487 mL | 1.6218 mL | |
| 60 mM | 0.0541 mL | 0.2703 mL | 0.5406 mL | 1.3515 mL | |
| 80 mM | 0.0405 mL | 0.2027 mL | 0.4055 mL | 1.0137 mL | |
| 100 mM | 0.0324 mL | 0.1622 mL | 0.3244 mL | 0.8109 mL |