Physicochemical Properties
| Molecular Formula | C10H12N4NA3O14P3 |
| Molecular Weight | 574.1113 |
| Exact Mass | 573.925 |
| CAS # | 35908-31-7 |
| PubChem CID | 135742691 |
| Appearance | Typically exists as White to off-white solids at room temperature |
| Density | 2.63g/cm3 |
| Boiling Point | 994.9ºC at 760mmHg |
| Flash Point | 555.5ºC |
| Hydrogen Bond Donor Count | 4 |
| Hydrogen Bond Acceptor Count | 16 |
| Rotatable Bond Count | 8 |
| Heavy Atom Count | 34 |
| Complexity | 862 |
| Defined Atom Stereocenter Count | 4 |
| SMILES | P(=O)([O-])(OP(=O)([O-])OP(=O)([O-])O[H])OC([H])([H])[C@]1([H])C([H])([C@@]([H])([C@]([H])(N2C([H])=NC3C(N([H])C([H])=NC2=3)=O)O1)O[H])O[H].[Na+].[Na+].[Na+] |
| InChi Key | QRGLCGLOQVQVCS-MSQVLRTGSA-K |
| InChi Code | InChI=1S/C10H15N4O14P3.3Na/c15-6-4(1-25-30(21,22)28-31(23,24)27-29(18,19)20)26-10(7(6)16)14-3-13-5-8(14)11-2-12-9(5)17;;;/h2-4,6-7,10,15-16H,1H2,(H,21,22)(H,23,24)(H,11,12,17)(H2,18,19,20);;;/q;3*+1/p-3/t4-,6-,7-,10-;;;/m1.../s1 |
| Chemical Name | trisodium;[[[(2R,3S,4R,5R)-3,4-dihydroxy-5-(6-oxo-1H-purin-9-yl)oxolan-2-yl]methoxy-oxidophosphoryl]oxy-oxidophosphoryl] hydrogen phosphate |
| Synonyms | Inosine-5'-triphosphate trisodium salt; 35908-31-7; Inosine 5' Triphosphate Trisodium Salt; Inosine 5'-triphosphate trisodium salt; MFCD00084678; trisodium;[[[(2R,3S,4R,5R)-3,4-dihydroxy-5-(6-oxo-1H-purin-9-yl)oxolan-2-yl]methoxy-oxidophosphoryl]oxy-oxidophosphoryl] hydrogen phosphate; ITP; Inosine-5'-triphosphatetrisodiumsalt; |
| 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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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 |
ATPases/GTPases - Nonselective adenosine receptors [1] - Nonselective adenosine receptors (mediates reno-protective effects via regulating IL-18, ICAM-1, and JNK-MAPK pathway) [2] |
| ln Vitro | G-proteins mediate signal transfer from receptors to effector systems. In their guanosine 5'-triphosphate (GTP)-bound form, G-protein alpha-subunits activate effector systems. Termination of G-protein activation is achieved by the high-affinity GTPase [E.C. 3.6.1.-] of their alpha-subunits. Like GTP, inosine 5'-triphosphate (ITP) and xanthosine 5'-triphosphate (XTP) can support effector system activation. We studied the interactions of GTP, ITP, and XTP with the retinal G-protein, transducin (TD), and with G-proteins in HL-60 leukemia cell membranes. TD hydrolyzed nucleoside 5'-triphosphates (NTPs) in the order of efficacy GTP > ITP > XTP. NTPs eluted TD from rod outer segment disk membranes in the same order of efficacy. ITP and XTP competitively inhibited TD-catalyzed GTP hydrolysis. In HL-60 membranes, the chemoattractants N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP) and leukotriene B4 (LTB4) effectively activated GTP and ITP hydrolysis by Gi-proteins. fMLP and LTB4 were at least 10-fold more potent activators of ITPase than of GTPase. Complement C5a effectively activated the GTPase of Gi-proteins but was only a weak stimulator of ITPase. The potency of C5a to activate GTP and ITP hydrolysis was similar. The fMLP-stimulated GTPase had a lower Km value than the fMLP-stimulated ITPase, whereas the opposite was true for the Vmax values. fMLP, C5a, and LTB4 did not stimulate XTP hydrolysis. Collectively, our data show that GTP, ITP, and XTP bind to G-proteins with different affinities, that G-proteins hydrolyze NTPs with different efficacies, and that chemoattractants stimulate GTP and ITP hydrolysis by Gi-proteins in a receptor-specific manner. On the basis of our results and the data in the literature, we put forward the hypothesis that GTP, ITP, and XTP act as differential signal amplifiers and signal sorters at the G-protein level[3]. |
| ln Vivo |
- In male Wistar rats trained for intravenous cocaine self-administration (0.6 mg/kg/infusion, Fixed Ratio 5 schedule), administration of 5'-N-Ethylcarboxamidoadenosine reduced the number of cocaine infusions per session compared to vehicle; this effect was primarily due to a marked increase in the latency for the first cocaine infusion, while responding after drug-induced delays remained at control levels [1] - In male Sprague-Dawley rats with streptozotocin-induced diabetes (6 weeks post-induction), daily intraperitoneal (i.p.) injections of 5'-N-Ethylcarboxamidoadenosine at 0.3 mg/kg/day for 2 weeks exerted hypoglycemic and antioxidant effects, alleviated diabetes-induced renal morphological changes (mild glomerular effects and reduced tubular epithelium vacuolation), and downregulated gene expression of IL-18, TNF-α, ICAM-1 as well as deactivated JNK-MAPK pathway [2] |
| Enzyme Assay | The methylation-dependent restriction endonuclease McrBC from Escherichia coli K12 cleaves DNA containing two R(m)C dinucleotides separated by about 40 to 2000 base-pairs. McrBC is unique in that cleavage is totally dependent on GTP hydrolysis. McrB is the GTP binding and hydrolyzing subunit, whereas MrC stimulates its GTP hydrolysis. The C-terminal part of McrB contains the sequences characteristic for GTP-binding proteins, consisting of the GxxxxGK(S/T) motif (position 201-208), followed by the DxxG motif (position 300-303). The third motif (NKxD) is present only in a non-canonical form (NTAD 333-336). Here we report a mutational analysis of the putative GTP-binding domain of McrB. Amino acid substitutions were initially performed in the three proposed GTP-binding motifs. Whereas substitutions in motif 1 (P203V) and 2 (D300N) show the expected, albeit modest effects, mutation in the motif 3 is at variance with the expectations. Unlike the corresponding EF-Tu and ras -p21 variants, the D336N mutation in McrB does not change the nucleotide specificity from GTP to XTP, but results in a lack of GTPase stimulation by McrC. The finding that McrB is not a typical G protein motivated us to perform a search for similar sequences in DNA databases. Eight microbial sequences were found, mainly from unfinished sequencing projects, with highly conserved sequence blocks within a presumptive GTP-binding domain. From the five sequences showing the highest homology, 17 invariant charged or polar residues outside the classical three GTP-binding motifs were identified and subsequently exchanged to alanine. Several mutations specifically affect GTP affinity and/or GTPase activity. Our data allow us to conclude that McrB is not a typical member of the superfamily of GTP-binding proteins, but defines a new subfamily within the superfamily of GTP-binding proteins, together with similar prokaryotic proteins of as yet unidentified function[2]. |
| Animal Protocol |
- Cocaine self-administration experiment (rats): Male Wistar rats were trained to obtain intravenous cocaine infusions (0.6 mg/kg/infusion) under a Fixed Ratio 5 schedule. 5'-N-Ethylcarboxamidoadenosine was administered, and the number of cocaine infusions per session and latency for the first infusion were recorded [1] - Diabetic nephropathy experiment (rats): Male Sprague-Dawley rats were induced with diabetes via streptozotocin. Six weeks later, rats received daily i.p. injections of 5'-N-Ethylcarboxamidoadenosine at 0.3 mg/kg/day for 2 weeks. After treatment, renal tissues were collected to assess morphological changes, malondialdehyde (oxidative stress marker), gene expression of IL-18/TNF-α/ICAM-1 (real-time PCR), and JNK-MAPK activation [2] |
| References |
[1]. Energy-transducing adenosine triphosphatase from Escherichia coli: purification, properties, and inhibition by antibody. J Bacteriol. 1973 May;114(2):772-81. [2]. The GTP-binding domain of McrB: more than just a variation on a common theme? J Mol Biol. 1999 Sep 24;292(3):547-56. [3]. Functionally nonequivalent interactions of guanosine 5'-triphosphate, inosine 5'-triphosphate, and xanthosine 5'-triphosphate with the retinal G-protein, transducin, and with Gi-proteins in HL-60 leukemia cell membranes. Biochem Pharmacol. 1997 Sep 1;54(5):551-62. [4]. te specificity of T4 RNA-ligase: the role of a purine nucleotide base in forming a covalent AMP-RNA-ligase complex. Biokhimiia. 1993 Jun;58(6):857-65. |
| Additional Infomation |
The membrane adenosine triphosphatase (E.C. 3.6.1.3) from Escherichia coli has been solubilized with Triton X-100 and purified to near homogeneity. The purified enzyme has a sedimentation coefficient of 12.9S in a sucrose gradient, corresponding to a molecular weight of about 360,000. On electrophoresis in gels containing sodium dodecyl sulfate, it dissociates into subunits with apparent molecular weights of 60,000, 56,000, 35,000, and 13,000. The purified enzyme loses activity and breaks down into subunits when stored in the cold. Guanosine 5'-triphosphate and inosine 5'-triphosphate are alternative substrates. Ca(2+) and, to a small extent, Co(2+) or Ni(2+) will substitute for Mg(2+) in the reaction. The K(m) for Mg-adenosine triphosphate of the membrane-bound enzyme is 0.23 mM, and for the pure enzyme it is 0.29 mM. Azide is a noncompetitive inhibitor of both the membrane-bound enzyme and the pure enzyme. P(i) is a noncompetitive inhibitor of the solubilized enzyme. An antibody to the purified enzyme was obtained from rabbits. The antibody inhibits the solubilized enzyme and virtually all of the adenosine triphosphate hydrolysis by membranes from cells grown aerobically or anaerobically. The antibody also inhibits the adenosine triphosphate-stimulated pyridine nucleotide transhydrogenase (E.C. 1.6.1.1) of the E. coli membrane.[1] - 5'-N-Ethylcarboxamidoadenosine is a nonselective adenosine agonist; its inhibition of cocaine self-administration may involve motivational systems regulation, with potential contribution from sedative effects (a known property of adenosine agonists) [1] - 5'-N-Ethylcarboxamidoadenosine is a stable nonselective adenosine agonist (avoids rapid extracellular adenosine degradation); its reno-protective effects support adenosine receptor activation as a potential target for diabetic nephropathy [2] |
Solubility Data
| Solubility (In Vitro) |
H2O : ~100 mg/mL (~174.18 mM) DMSO :< 1 mg/mL |
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.7418 mL | 8.7091 mL | 17.4183 mL | |
| 5 mM | 0.3484 mL | 1.7418 mL | 3.4837 mL | |
| 10 mM | 0.1742 mL | 0.8709 mL | 1.7418 mL |