| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| Other Sizes |
| Targets |
IC50: 22 nM (PDE9A)[1]
Phosphodiesterase 9A (PDE9A). PDE9A is a member of the phosphodiesterase superfamily that specifically hydrolyzes cyclic guanosine monophosphate (cGMP) to its inactive 5-prime form, 5'-GMP. By inhibiting PDE9A, PF-04449613 prevents the degradation of cGMP, leading to elevated intracellular cGMP levels and activation of downstream signaling pathways involved in synaptic plasticity and motor learning. |
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| ln Vitro |
PF-04449613 selectively inhibits PDE9A with an IC50 of 22 nM. In vitro, the compound promotes dendritic spine formation in cultured neurons. As a highly selective PDE9A inhibitor, it shows minimal activity against other PDE family members, reducing off-target effects. The (S)-enantiomer is the more active isomer, with the (R)-isomer showing reduced or negligible activity at the target. (S)-PF-04449613 is the active enantiomer used for in vitro and in vivo studies.
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| ln Vivo |
In vivo, PF-04449613 improves motor learning ability in a mouse model. The compound also promotes dendritic spine formation and performance improvement after motor learning. These findings suggest that PDE9A inhibition enhances synaptic plasticity and learning-related behaviors, making (S)-PF-04449613 a valuable tool for studying the role of cGMP signaling in learning, memory, and neurodegenerative diseases such as Alzheimer's disease and Huntington's disease.
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| Enzyme Assay |
A PDE9A enzyme inhibition assay is performed in 384-well format using purified recombinant human PDE9A enzyme. The enzyme is incubated with [3H]-cGMP (10-20 nM) and varying concentrations of PF-04449613 (0.001-100 uM) in assay buffer (50 mM Tris-HCl, pH 7.5, 8.3 mM MgCl2, 1.7 mM EDTA) at room temperature for 20-60 minutes. The reaction is terminated by adding scintillation proximity assay (SPA) beads, and radioactivity is measured. IC50 values are calculated using a four-parameter logistic curve fit. For cell-based PDE inhibition assays, HEK293 cells expressing PDE9A are seeded in 96-well plates and treated with serial dilutions of PF-04449613 (0.1 nM to 10 uM) for 30-60 minutes. After stimulation with a nitric oxide donor or guanylyl cyclase activator to increase cGMP production, cells are lysed, and cGMP levels are measured by homogeneous time-resolved fluorescence (HTRF) or competitive ELISA. The EC50 for cGMP elevation is calculated.
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| Cell Assay |
For in vivo efficacy in motor learning, C57BL/6 mice are administered PF-04449613 orally or intraperitoneally at doses of 1-30 mg/kg. Motor learning is assessed using the accelerating rotarod test: mice are placed on a rotating rod that accelerates from 4 to 40 rpm over 5 minutes. Latency to fall is recorded over multiple training sessions. Brain tissue is collected at endpoint for analysis of cGMP levels by LC-MS/MS and for Golgi staining to assess dendritic spine density in motor cortex and striatum.
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| Animal Protocol |
PF-04449613 is an orally bioavailable and brain-penetrant PDE9A inhibitor. In preclinical species, the compound shows moderate to high oral bioavailability (F% = 30-70%), moderate plasma clearance, and a terminal half-life suitable for once- or twice-daily dosing. Brain-to-plasma ratios indicate good central nervous system penetration. The compound is soluble in DMSO at 100 mg/mL and can be formulated for in vivo studies using vehicles such as PEG400/saline or DMSO/PEG300/Tween-80/saline. Storage: -20degC powder; -80degC in solvent for 6 months.
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| ADME/Pharmacokinetics |
In preclinical toxicology studies, PDE9A inhibitors are generally well-tolerated at therapeutic doses. PF-04449613 has not shown significant toxicity in animal models at doses up to 30 mg/kg. As a cGMP-specific PDE inhibitor, potential adverse effects may include mild gastrointestinal disturbances (nausea, diarrhea) and vasodilation-related effects (hypotension, headache). No significant hepatotoxicity, nephrotoxicity, or cardiotoxicity (hERG inhibition) has been reported. However, comprehensive toxicology data is not publicly available.
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| Toxicity/Toxicokinetics |
PF-04449613 was discovered and characterized by researchers at Pfizer through structure-based drug design and parallel chemistry. The compound is a selective PDE9A inhibitor that promotes dendritic spine formation and performance improvement after motor learning. (S)-PF-04449613 is the more active enantiomer used for pharmacological studies. Key references include Claffey MM, et al. J Med Chem. 2012;55(21):9055-9068 and Lai B, et al. Dev Neurobiol. 2018;78(9):859-872. This product is for research use only and is not FDA-approved for human therapy.
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| References |
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| Molecular Formula |
C21H25N5O3
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| Molecular Weight |
395.45
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| Related CAS # |
PF-04449613;1236858-52-8
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| Appearance |
White to light yellow solid powder
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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 |
| 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) |
DMSO :~100 mg/mL (~252.88 mM)
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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 | 2.5288 mL | 12.6438 mL | 25.2876 mL | |
| 5 mM | 0.5058 mL | 2.5288 mL | 5.0575 mL | |
| 10 mM | 0.2529 mL | 1.2644 mL | 2.5288 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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