| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
PKCα 5.6 nM (IC50) PKCβI 14.5 nM (IC50) PKCβII 13 nM (IC50) PKCγ 37.7 nM (IC50) PKCθ 74.1 nM (IC50)
PF-03622905 targets protein kinase C (PKC) isoforms, including PKCα, PKCβI, PKCβII, PKCγ, and PKCθ. It binds the ATP-binding site of the PKC kinase domain and competitively inhibits ATP binding. This blocks PKC-mediated phosphorylation of downstream substrates. PKCβII activation in hyperglycemic states is implicated in diabetic microvascular abnormalities, making PKC a therapeutic target for diabetic retinopathy and retinal vascular leakage. |
|---|---|
| ln Vitro |
PF-03622905 inhibits phospho-ERK1/2 creation, phospho-SHP2 levels, and interleukin 8 release in a concentration-dependent manner; the IC50 values for these reactions are 0.15 μM, 0.16 μM, and 35 nM, respectively[1]. When applied at 1 μM to human umbilical vein endothelial cells, PF-03622905 exhibits minimal cytotoxicity, leaving 83.3% of the cells viable[1].
In vitro, PF-03622905 inhibits PKCα with an IC50 of 5.6 nM, PKCβI with 14.5 nM, PKCβII with 13 nM, PKCγ with 37.7 nM, and PKCθ with 74.1 nM. It shows high specificity for PKC over other protein kinases. In cellular assays, it causes concentration-dependent inhibition of phospho-ERK1/2 formation (IC50 = 0.15 µM), interleukin-8 release (IC50 = 0.16 µM), and phospho-SHP2 levels (IC50 = 35 nM). Low cytotoxicity is observed at 1 µM against HUVECs, with 83.3% viability. |
| ln Vivo |
In vivo, PF-03622905 demonstrates efficacy in ameliorating retinal vascular leakage in a diabetic rat model. When administered orally, it shows good exposure in the retina and reverses vascular leakage in streptozotocin-induced diabetic rats. This in vivo validation confirms its potential as a therapeutic agent for diabetic retinopathy. The compound's ability to block PKC signaling in retinal tissue underlies its efficacy in reducing vascular permeability and microvascular complications.
|
| Enzyme Assay |
For non-cell enzyme assays, recombinant PKC isoforms are incubated with PF-03622905 and a fluorescent or radiolabeled peptide substrate in kinase assay buffer containing ATP. Kinase activity is measured by fluorescence polarization or radiometric detection. IC50 values are calculated from dose-response curves. Selectivity is assessed against a panel of other protein kinases to confirm specificity for PKC over other kinase families.
|
| Cell Assay |
In cell-based functional assays, PF-03622905 is tested for inhibition of PKC-mediated signaling. It causes concentration-dependent inhibition of phospho-ERK1/2 formation, interleukin-8 release, and phospho-SHP2 levels. These readouts confirm functional PKC inhibition in cellular contexts. Cytotoxicity is assessed in human umbilical vein endothelial cells (HUVECs) at 1 µM, with cell viability remaining at 83.3%, indicating low toxicity at effective concentrations.
|
| Animal Protocol |
For in vivo animal studies, streptozotocin-induced diabetic Long-Evans rats are used. PF-03622905 is administered orally, and retinal exposure is assessed. Retinal vascular leakage is measured using Evans blue dye or fluorescein angiography. PKC activity in retinal lysates is measured by kinase assay. The compound shows good retinal exposure and ameliorates vascular leakage, demonstrating target engagement and efficacy in the diabetic retina.
|
| References |
| Molecular Formula |
C24H35N7O3
|
|---|---|
| Molecular Weight |
469.57980465889
|
| Exact Mass |
469.28
|
| CAS # |
1072100-15-2
|
| PubChem CID |
66738800
|
| Appearance |
Typically exists as solid at room temperature
|
| LogP |
1
|
| tPSA |
118
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
6
|
| Heavy Atom Count |
34
|
| Complexity |
735
|
| Defined Atom Stereocenter Count |
2
|
| SMILES |
CCCN1C[C@@H](N(C[C@H]1CCO)C(=O)N2CC3=C(C2(C)C)NN=C3NC(=O)C4=CC=CC=N4)C
|
| InChi Key |
SYMKCZOALSNIBQ-DLBZAZTESA-N
|
| InChi Code |
InChI=1S/C24H35N7O3/c1-5-11-29-13-16(2)30(14-17(29)9-12-32)23(34)31-15-18-20(24(31,3)4)27-28-21(18)26-22(33)19-8-6-7-10-25-19/h6-8,10,16-17,32H,5,9,11-15H2,1-4H3,(H2,26,27,28,33)/t16-,17+/m0/s1
|
| Chemical Name |
N-[5-[(2S,5R)-5-(2-hydroxyethyl)-2-methyl-4-propylpiperazine-1-carbonyl]-6,6-dimethyl-1,4-dihydropyrrolo[3,4-c]pyrazol-3-yl]pyridine-2-carboxamide
|
| 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
|
|---|---|
| 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 | 2.1296 mL | 10.6478 mL | 21.2956 mL | |
| 5 mM | 0.4259 mL | 2.1296 mL | 4.2591 mL | |
| 10 mM | 0.2130 mL | 1.0648 mL | 2.1296 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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