Physicochemical Properties
| Molecular Formula | C10H14N5O7P |
| Molecular Weight | 347.2212 |
| Exact Mass | 365.073 |
| CAS # | 18422-05-4 |
| Related CAS # | Adenosine monophosphate;61-19-8;Adenosine monophosphate-13C10,15N5 disodium;Adenosine 5'-monophosphate disodium;4578-31-8;Adenosine 5'-monophosphate-13C disodium;Adenosine 5'-monophosphate-d2 disodium;Adenosine-5'-monophosphate-15N5 disodium |
| PubChem CID | 6419976 |
| Appearance | White to off-white solid powder |
| Boiling Point | 798.5ºC at 760mmHg |
| Melting Point | 183-188 °C (dec.)(lit.) |
| Flash Point | 436.7ºC |
| Vapour Pressure | 6.7E-27mmHg at 25°C |
| Hydrogen Bond Donor Count | 6 |
| Hydrogen Bond Acceptor Count | 12 |
| Rotatable Bond Count | 4 |
| Heavy Atom Count | 24 |
| Complexity | 481 |
| Defined Atom Stereocenter Count | 4 |
| SMILES | C1=NC(=C2C(=N1)N(C=N2)[C@H]3[C@@H]([C@@H]([C@H](O3)COP(=O)(O)O)O)O)N.O |
| InChi Key | ZOEFQKVADUBYKV-MCDZGGTQSA-N |
| InChi Code | InChI=1S/C10H14N5O7P.H2O/c11-8-5-9(13-2-12-8)15(3-14-5)10-7(17)6(16)4(22-10)1-21-23(18,19)20;/h2-4,6-7,10,16-17H,1H2,(H2,11,12,13)(H2,18,19,20);1H2/t4-,6-,7-,10-;/m1./s1 |
| Chemical Name | [(2R,3S,4R,5R)-5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methyl dihydrogen phosphate;hydrate |
| 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 5'-monophosphate monohydrate is an A1 receptor agonist [1]. Adenosine 5'-monophosphate monohydrate (5'-AMP) at doses ranging from 25 to 400 μM caused minimal cytotoxicity in RAW264.7 cells. Adenosine 5'-monophosphate monohydrate dramatically reduced TNF-α and IL-6 mRNA expression in RAW264.7 cells. At 400 μM, adenosine 5'-monophosphate monohydrate had the strongest inhibitory effect on TNF-α and IL-6 mRNA levels. Exposing cells to adenosine 5'-monophosphate monohydrate decreases NF-κB p65 recruitment to TNF-α, IL-6, and IL-1β transcription promoters [2]. |
| ln Vitro |
Adenosine 5'-monophosphate monohydrate is an A1 receptor agonist [1]. Adenosine 5'-monophosphate monohydrate (5'-AMP) at doses ranging from 25 to 400 μM caused minimal cytotoxicity in RAW264.7 cells. Adenosine 5'-monophosphate monohydrate dramatically reduced TNF-α and IL-6 mRNA expression in RAW264.7 cells. At 400 μM, adenosine 5'-monophosphate monohydrate had the strongest inhibitory effect on TNF-α and IL-6 mRNA levels. Exposing cells to adenosine 5'-monophosphate monohydrate decreases NF-κB p65 recruitment to TNF-α, IL-6, and IL-1β transcription promoters [2]. In Vitro: - AMP and its non-hydrolyzable analog deoxyadenosine 5‘-monophosphonate (ACP) directly activate the human adenosine A1 receptor (hA1R) in HEK293 and COS7 cells, as measured by chimeric G protein (Gqi)-mediated calcium mobilization. AMP and adenosine are equipotent at hA1R.[1] - AMP does not directly activate the human adenosine A2B receptor (hA2BR). Activation of hA2BR by AMP requires its prior hydrolysis to adenosine by ecto-5‘-nucleotidase (NT5E/CD73) or prostatic acid phosphatase (PAP).[1] - In mouse embryonic cortical neurons, ACP (a non-hydrolyzable AMP analog) inhibits forskolin-evoked cAMP accumulation via endogenous A1R, demonstrating direct activation of native A1R signaling pathways.[1] - The P2Y receptor antagonists pyridoxalphosphate-6-azophenyl-2‘,4‘-disulfonic acid and suramin do not block AMP-evoked calcium responses in hA1R-expressing cells, indicating that AMP signaling is specific to A1R and not mediated through P2Y receptors.[1] - Site-directed mutagenesis of the hA1R ligand-binding pocket (His-278 to Ala) abolishes activation by both adenosine and AMP, while mutation of His-251 to Ala specifically reduces AMP potency without affecting adenosine potency, providing molecular evidence for direct AMP binding to A1R.[1] |
| ln Vivo | Adenosine 5'-monophosphate monohydrate (5'-AMP) treatment dramatically raised the amounts of adenosine in the liver of C57BL/6J mice. In comparison, the survival rates of mice treated with adenosine 5'-monophosphate monohydrate (n=15) were 100% (8 h) and 93.3% (24 h), whereas the survival rates of mice treated with PBS (n=15) were 60% (8 h) and 33.3% (24 h). When comparing the adenosine 5'-monophosphate monohydrate group to the vehicle group, there was a substantial decrease in the levels of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT). In the 5'-adenosine monophosphate monohydrate group, there was a decrease in the area and severity of necrosis as well as a reduction in the infiltration of inflammatory cells [2]. |
| Enzyme Assay |
- Ectonucleotidase activity is assessed indirectly. To test if AMP activation of receptors is dependent on hydrolysis to adenosine, experiments are performed in the presence of the competitive NT5E inhibitor α,β-methylene-ADP (αβ-met-ADP, 10 µM). Cells are incubated with the inhibitor for 3 minutes prior to and during agonist stimulation.[1] - The persistence of AMP-evoked calcium responses in hA1R-expressing cells in the presence of αβ-met-ADP, and the lack of such responses in hA2BR-expressing cells under the same condition, is used to conclude direct versus ectonucleotidase-dependent activation.[1] |
| Cell Assay |
- Calcium Imaging Assay: HEK293 cells are grown on coated glass-bottom dishes and transfected with plasmids encoding human A1R (or A2BR) and a chimeric G protein (Gqi or Gqs). Transfected cells are identified via co-transfection with a Venus fluorescent protein plasmid. ~24 hours post-transfection, cells are loaded with the calcium indicator Fura-2 AM for 1 hour at room temperature. After washing, cells are imaged on a fluorescence microscope. Baseline fluorescence (340 nm/380 nm excitation ratio) is recorded for 40 seconds before manually adding agonist solutions (e.g., AMP, adenosine). Calcium mobilization is monitored in real-time, and responses are quantified by calculating the area under the curve (AUC) for 1 minute post-agonist addition.[1] - cAMP Accumulation Assay (GloSensor): HEK293T cells are co-transfected with a plasmid encoding hA1R and the GloSensor 22F cAMP reporter plasmid. After transfection, cells are incubated with test compounds (e.g., AMP, adenosine, CCPA) for 10 minutes, followed by stimulation with a submaximal concentration of the β-adrenergic agonist isoproterenol for 7 minutes to elevate cAMP levels. Following incubation, the GloSensor substrate reagent is added, and luminescence is measured. A1R activation inhibits isoproterenol-stimulated cAMP accumulation, which is detected as a decrease in luminescence.[1] - Primary Neuronal cAMP ELISA: Cortical neurons are dissected from mouse embryos and plated. After 1 day in vitro, neurons are pre-treated with compounds (e.g., ACP, CPA) for 30 minutes, then stimulated with forskolin for 15 minutes to activate adenylyl cyclase. Cells are then lysed, and intracellular cAMP levels are measured using a commercial ELISA kit, normalized to total protein content.[1] |
| References |
[1]. AMP is an adenosine A1 receptor agonist. J Biol Chem. 2012 Feb 17;287(8):5301-9. [2]. Adenosine 5'-monophosphate ameliorates D-galactosamine/lipopolysaccharide-induced liver injury through an adenosine receptor-independent mechanism in mice. Cell Death Dis. 2014 Jan 9;5:e985. [3]. Comparison of the antiviral effects of 5-methoxymethyldeoxyuridine-5'-monophosphate with adenine arabinoside-5'-monophosphate. Antiviral Res. 1983 Sep;3(3):161-74. |
| Additional Infomation |
Adenosine 5'-monophosphate monohydrate is a purine ribonucleoside monophosphate. - Background & Significance: Prior to this study, no definitive receptor for AMP was known. Its diverse physiological effects were often attributed to its hydrolysis to adenosine by ectonucleotidases. This study identifies the adenosine A1 receptor (A1R) as a bona fide direct receptor for AMP, suggesting that some physiological effects of AMP may be ectonucleotidase-independent.[1] - Mechanism: AMP acts as a full agonist at the A1R. Activation is direct and does not require conversion to adenosine. The negatively charged phosphate group of AMP likely interacts with specific histidine residues (e.g., His-251) in the ligand-binding pocket of A1R.[1] - Implications: This finding suggests that AMP-based prodrugs designed to target other adenosine receptors (e.g., A2AR) after hydrolysis may also have off-target direct effects on A1R. It also reinterprets the cardioprotective effects of some AMP analogs, which may be mediated by A1R rather than P2X receptors as previously hypothesized.[1] |
Solubility Data
| Solubility (In Vitro) | H2O : ~5 mg/mL (~13.69 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: 5.26 mg/mL (14.40 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication (<60°C). (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.8800 mL | 14.4001 mL | 28.8002 mL | |
| 5 mM | 0.5760 mL | 2.8800 mL | 5.7600 mL | |
| 10 mM | 0.2880 mL | 1.4400 mL | 2.8800 mL |