| Size | Price | Stock | Qty |
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| 2mg |
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| 50mg | |||
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EHNA HCl, the hydrochloride salt of EHNA, is a novel and potent inhibitor of adenosine deaminase, also an inhibitor of cGMP-stimulated phosphodiesterase (PDE2) with IC50 of 0.8 - 4 μM.
| Targets |
The target of EHNA hydrochloride (EHNA) is cyclic nucleotide phosphodiesterase 2 (PDE2), with a Ki value of 0.2 μM; it shows no significant inhibition on PDE1, PDE3, PDE4, or PDE5 (inhibition rate <10% at 10 μM) [1]
The target of EHNA hydrochloride (EHNA) is PDE2; it selectively inhibits PDE2 activity without affecting PDE4 activity (no inhibition observed at 10 μM) [2] |
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| ln Vitro |
While having a significantly lesser inhibitory effect on unstimulated PDE2 activity, EHNA totally removes cyclic GMP's capacity to trigger PDE2 activity [2]. The Hill plot is almost in accord, and EHNA shows typical Michael kinetic inhibition of cyclic GMP-stimulated PDE2 activity [2]. In fibroblasts, EHNA raises mitochondrial dATP levels and inhibits dAdo breakdown [3].
1. Isozyme-selective PDE inhibition (Reference [1]): EHNA hydrochloride (0.01-100 μM) was tested for inhibitory activity against various PDE isozymes. It potently inhibited PDE2 (Ki=0.2 μM) and weakly inhibited cGMP-stimulated PDE activity in rat brain homogenates (IC50=0.5 μM). At 10 μM, it inhibited PDE2 by >90% but had <10% inhibition on PDE1 (bovine heart), PDE3 (rabbit heart), PDE4 (human monocytes), and PDE5 (rat lung) [1] 2. Regulation of PDE activity in thymocytes (Reference [2]): Mouse thymocytes were stimulated with anti-CD3 antibody (T cell antigen receptor ligation) for 0-60 minutes. Pretreatment with EHNA hydrochloride (10 μM) for 30 minutes completely blocked the stimulation-induced increase in PDE2 activity (which normally rose by 2.3-fold at 30 minutes). PDE4 activity was not affected by EHNA hydrochloride, remaining unchanged compared to the stimulated control [2] 3. cAMP level modulation (Reference [2]): Thymocytes treated with EHNA hydrochloride (10 μM) + anti-CD3 antibody had a 1.8-fold higher intracellular cAMP level than those treated with anti-CD3 alone. This was attributed to PDE2 inhibition, as PDE2 normally degrades cAMP in thymocytes [2] |
| Enzyme Assay |
1. PDE activity assay (Reference [1]): Prepare reaction mixtures containing purified PDE isozymes (PDE1-PDE5) or rat brain homogenates, 0.5 μM [³H]-cGMP (for cGMP-stimulated PDE) or [³H]-cAMP (for cAMP-specific PDE), and EHNA hydrochloride (0.01-100 μM). Incubate at 37°C for 15 minutes. Terminate the reaction by adding snake venom phosphodiesterase and incubate for another 10 minutes. Separate [³H]-adenosine/guanosine from unreacted [³H]-cAMP/cGMP using anion-exchange resin. Measure radioactivity with a scintillation counter to calculate PDE activity and inhibition rate [1]
2. Thymocyte PDE activity assay (Reference [2]): Mouse thymocytes were lysed in hypotonic buffer and centrifuged at 100,000×g for 60 minutes to obtain the cytosolic fraction. The fraction was mixed with EHNA hydrochloride (0-10 μM), 1 μM [³H]-cAMP, and PDE reaction buffer (containing MgCl₂ and Tris-HCl, pH 7.5). Incubate at 37°C for 20 minutes. Terminate and quantify as described in Reference [1]. PDE2 activity was calculated by subtracting PDE activity in the presence of 10 μM EHNA hydrochloride (complete PDE2 inhibition) from total PDE activity [2] |
| Cell Assay |
1. Thymocyte stimulation and PDE activity detection (Reference [2]): Isolate mouse thymocytes and suspend them in RPMI 1640 medium. Divide into three groups: (1) Control (no treatment); (2) Anti-CD3 group (10 μg/mL anti-CD3 antibody, stimulation for 0-60 minutes); (3) EHNA hydrochloride + anti-CD3 group (10 μM EHNA hydrochloride pretreatment for 30 minutes, then 10 μg/mL anti-CD3 for 0-60 minutes). After stimulation, lyse cells to extract cytosolic fractions. Measure PDE2 and PDE4 activities using the enzyme assay described above. Intracellular cAMP levels were measured by radioimmunoassay (RIA) with a [¹²⁵I]-cAMP kit [2]
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| References |
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| Additional Infomation |
(2R,3S)-EHNA hydrochloride is a hydrochloride prepared by reacting (2R,3S)-EHNA with an equivalent amount of hydrochloric acid. It is a selective inhibitor of cGMP-stimulated phosphodiesterase (PDE2) (IC50 = 0.8-4 mM) and a potent inhibitor of adenosine deaminase. It also functions as an inhibitor of EC 3.1.4 (phosphodiester hydrolase) and EC 3.5.4.4 (adenosine deaminase). It contains (2R,3S)-EHNA(1+).
1. Chemical composition (References [1][2]): EHNA hydrochloride is the hydrochloride of erythro-9-(2-hydroxy-3-nonyl)adenine, a synthetic purine derivative commonly used as a selective PDE2 inhibitor in research [1][2]. 2. Mechanism of action (References [1][2]): EHNA hydrochloride works by binding to the active site of PDE2, inhibiting the enzyme's ability to hydrolyze cyclic nucleotides (cAMP and cGMP). This leads to an increase in intracellular cAMP/cGMP levels, thereby regulating signaling pathways involved in cell proliferation, differentiation, and immune responses (e.g., T cell activation in thymocytes) [1][2] 3. Research applications (References [2]): EHNA hydrochloride is widely used as a tool compound to distinguish the role of PDE2-mediated responses from those of other PDE isoenzymes (e.g., PDE4) in cell and molecular biology studies, particularly in the study of cyclic nucleotide signaling in immune cells [2] |
| Molecular Formula |
C14H23N5O.HCL
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|---|---|
| Molecular Weight |
313.82626
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| Exact Mass |
313.166
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| CAS # |
58337-38-5
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| PubChem CID |
11957547
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| Appearance |
White to light yellow solid powder
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| Density |
1.27g/cm3
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| Boiling Point |
478.2ºC at 760mmHg
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| Flash Point |
243ºC
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| LogP |
3.684
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
7
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| Heavy Atom Count |
21
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| Complexity |
291
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| Defined Atom Stereocenter Count |
2
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| SMILES |
CCCCCC[C@@H]([C@@H](C)O)N1C=NC2=C(N=CN=C21)N.Cl
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| InChi Key |
VVDXNJRUNJMYOZ-DHXVBOOMSA-N
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| InChi Code |
InChI=1S/C14H23N5O.ClH/c1-3-4-5-6-7-11(10(2)20)19-9-18-12-13(15)16-8-17-14(12)19;/h8-11,20H,3-7H2,1-2H3,(H2,15,16,17);1H/t10-,11+;/m1./s1
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| Chemical Name |
(2R,3S)-3-(6-aminopurin-9-yl)nonan-2-ol;hydrochloride
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| HS Tariff Code |
2934.99.9001
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| 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)
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| Solubility (In Vitro) |
H2O : ~100 mg/mL (~318.64 mM)
DMSO : ~83.33 mg/mL (~265.53 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (6.63 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 20.8 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: ≥ 2.08 mg/mL (6.63 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 20.8 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. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (6.63 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.1864 mL | 15.9322 mL | 31.8644 mL | |
| 5 mM | 0.6373 mL | 3.1864 mL | 6.3729 mL | |
| 10 mM | 0.3186 mL | 1.5932 mL | 3.1864 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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