| ln Vitro |
PDE4B/D-IN-5 effectively inhibited recombinant PDE4B1 (IC50 = 3.4 nM) in in vitro fluorescent polarizoase activity assays [1]. PDE4B/D-IN-5 (15 μM; duration of action up to 90 minutes) showed high metabolic stability in rat liver microsomes, with 94.8% remaining after 60 minutes, a half-life of 5.5 hours, and a clearance rate as low as 0.0021 mL min−1 mg−1 [1]. PDE4B/D-IN-5 (1-4 μM; duration of action 1 hour) inhibited LPS-stimulated TNF-α production in RAW264.7 mouse macrophages in a dose-dependent manner [1].
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|---|---|
| ln Vivo |
PDE4B/D-IN-5 (1–6 mg/kg; orally; single dose) did not induce vomiting in beagle dogs [1]. PDE4B/D-IN-5 (6–12 mg/kg; intraperitoneally; daily; 5 days) reduced LPS-induced acute lung injury in male C57BL/6 mice in a dose-dependent manner through anti-inflammatory, antioxidant, and anti-apoptotic effects mediated by the cAMP-Epac pathway [1].
|
| Cell Assay |
ELISA detection [1]
Cell Types: RAW264.7 mouse macrophages Tested Concentrations: 1-4 μM Incubation Duration: 1 hour Experimental Results: Compared with the LPS-induced model group (TNF-α level was 26 pg/mL, while the baseline level of untreated cells was 9 pg/mL), the TNF-α level in the supernatant decreased in a dose-dependent manner. |
| Animal Protocol |
Animal/Disease Models:C57BL/6 mice (male, 8 weeks old, acute lung injury model established by intratracheal instillation of LPS) [1]
Doses: 6 mg/kg; 12 mg/kg Route of Administration: Intraperitoneal injection; once daily for 5 consecutive days Experimental Results: Lung erythema, lung enlargement, alveolar damage, interstitial edema and inflammatory cell infiltration were all reduced. Compared with the model group, the Smith lung injury score of the 12 mg/kg group was significantly reduced. The lung wet weight/dry weight ratio decreased in a dose-dependent manner. The neutrophil count in bronchoalveolar lavage fluid (BALF) was more effective than that of dexamethasone in a dose-dependent manner. Compared with dexamethasone, this drug can more effectively inhibit the activity of pulmonary myeloperoxidase (MPO) in a dose-dependent manner. This drug can reduce lung tissue nitric oxide (NO) levels to levels comparable to dexamethasone. It inhibits the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) in bronchoalveolar lavage fluid (BALF) in a dose-dependent manner, with the 12 mg/kg group showing stronger inhibitory effects than the dexamethasone group. It increases lung tissue cAMP levels in a dose-dependent manner, with the 12 mg/kg group showing the most significant effect. It significantly upregulates Epac1 protein expression in a dose-dependent manner. It significantly reduces the Bax/Bcl2 ratio, indicating inhibition of mitochondrial apoptosis. The drug has no statistically significant effect on the p-PKA/PKA or p-AMPK/AMPK ratios. |
| References |
| Molecular Formula |
C22H21F2N5O2
|
|---|---|
| Molecular Weight |
425.43
|
| CAS # |
3064814-26-9
|
| Appearance |
Typically exists as solids at room temperature
|
| SMILES |
FC([H])(CN1N=C(C2=CC=NN2CC)C3=C1C=CC(NC4=CC=C(CC(O)=O)C=C4)=C3)F
|
| 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)
|
| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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|---|---|
| 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
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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). View More
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)] 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). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3506 mL | 11.7528 mL | 23.5056 mL | |
| 5 mM | 0.4701 mL | 2.3506 mL | 4.7011 mL | |
| 10 mM | 0.2351 mL | 1.1753 mL | 2.3506 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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