1. GPCR/G Protein
  2. Adenosine Receptor
  3. 5'-N-Ethylcarboxamidoadenosine

5'-N-Ethylcarboxamidoadenosine  (Synonyms: NECA)

Cat. No.: HY-103173 Purity: 99.86%
SDS COA Handling Instructions

5'-N-Ethylcarboxamidoadenosine (NECA) is a nonselective adenosine receptor agonist.

For research use only. We do not sell to patients.

5'-N-Ethylcarboxamidoadenosine Chemical Structure

5'-N-Ethylcarboxamidoadenosine Chemical Structure

CAS No. : 35920-39-9

Size Price Stock Quantity
Solid + Solvent (Highly Recommended)
10 mM * 1 mL in DMSO
ready for reconstitution
USD 73 In-stock
Solution
10 mM * 1 mL in DMSO USD 73 In-stock
Solid
5 mg USD 50 In-stock
10 mg USD 63 In-stock
25 mg USD 120 In-stock
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Customer Review

Based on 6 publication(s) in Google Scholar

Top Publications Citing Use of Products
  • Biological Activity

  • Protocol

  • Purity & Documentation

  • References

  • Customer Review

Description

5'-N-Ethylcarboxamidoadenosine (NECA) is a nonselective adenosine receptor agonist.

IC50 & Target

Adenosine receptor[1]

Cellular Effect
Cell Line Type Value Description References
CHO EC50
> 10000 nM
Compound: 1b, NECA
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
[PMID: 25780876]
CHO EC50
0.018 μM
Compound: 2 (NECA)
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
[PMID: 23245803]
CHO EC50
0.04 μM
Compound: NECA
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
[PMID: 11784146]
CHO EC50
10.2 nM
Compound: NECA
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
[PMID: 21858244]
CHO EC50
12.58 nM
Compound: 1b, NECA
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
[PMID: 25780876]
CHO EC50
30 nM
Compound: NECA
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
[PMID: 20031406]
CHO EC50
38 nM
Compound: NECA
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
[PMID: 20031406]
CHO EC50
45 nM
Compound: NECA
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
10.1039/C5MD00086F
CHO EC50
63 nM
Compound: NECA
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
[PMID: 20031406]
CHO-K1 EC50
17.6 nM
Compound: 2, NECA
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
[PMID: 24900277]
COS-7 EC50
0.078 μM
Compound: NECA
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
[PMID: 16640329]
COS-7 EC50
10.4 μM
Compound: NECA
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
[PMID: 16640329]
HCT-116 IC50
> 10 μM
Compound: NECA
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
[PMID: 35707146]
HEK293 EC50
188 nM
Compound: 37; NECA
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
[PMID: 32271569]
HEK293 IC50
3.7 x 10-2 μM
Compound: NECA
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
[PMID: 27876250]
PBMC IC50
187 nM
Compound: NECA
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
[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
[PMID: 20031406]
PC-12 EC50
130 nM
Compound: 1
Potency against PC12 cell A2 adenosine receptor by adenylate cyclase activation
Potency against PC12 cell A2 adenosine receptor by adenylate cyclase activation
[PMID: 3018244]
PC-3 IC50
> 10 μM
Compound: NECA
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
[PMID: 35707146]
WI-38 IC50
> 10 μM
Compound: NECA
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
[PMID: 35707146]
In Vivo

After the administration of 5'-N-Ethylcarboxamidoadenosine (NECA), the mean number of cocaine infusions obtained per session is decreased significantly in a dose-dependent manner [5'-N-Ethylcarboxamidoadenosine (NECA): F(4,12)=14.9; P<0.001]. The administration of 5'-N-Ethylcarboxamidoadenosine (NECA) [F(4,12)=16.1; P<0.001] results in a significant increase in latencies above values obtained for vehicle treatment[1]. Daily i.p. injection of 5'-N-Ethylcarboxamidoadenosine (NECA) at 0.3 mg/kg/day for two weeks reduces malondialdehyde (MDA) levels in diabetic rats, but does not affect control rats. Daily treatment with NECA (0.3 mg/kg/day, i.p. for two weeks) reduces diabetes-induced gene expression of tumor necrosis factor (TNF)-α and interleukin (IL)-18 in diabetic rats, but does not affect control rats. Daily i.p. injection of 5'-N-Ethylcarboxamidoadenosine (NECA) at 0.3 mg/kg/day for two weeks also blocks the activation of JNK MAPK in diabetic rats, but does not affect control rats[2].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Molecular Weight

308.29

Formula

C12H16N6O4

CAS No.
Appearance

Solid

Color

White to off-white

SMILES

O[C@H]1[C@H](N2C=NC3=C(N)N=CN=C23)O[C@H](C(NCC)=O)[C@H]1O

Shipping

Room temperature in continental US; may vary elsewhere.

Storage
Powder -20°C 3 years
4°C 2 years
In solvent -80°C 2 years
-20°C 1 year
Solvent & Solubility
In Vitro: 

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)

Preparing
Stock Solutions
Concentration Solvent Mass 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
View the 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.

  • Molarity Calculator

  • Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Mass
=
Concentration
×
Volume
×
Molecular Weight *

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

C1

×
Volume (start)

V1

=
Concentration (final)

C2

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Volume (final)

V2

In Vivo:

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.

  • Protocol 1

    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.
  • Protocol 2

    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.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

g

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
DMSO +
+
%
Tween-80 +
%
Saline
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
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).
Calculation results:
Working solution concentration: mg/mL
Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).
The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
 If the continuous dosing period exceeds half a month, please choose this protocol carefully.
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.
Purity & Documentation

Purity: 99.86%

References
Animal Administration
[1]

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].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

References

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
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Product Name:
5'-N-Ethylcarboxamidoadenosine
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HY-103173
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