| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
PKCα 2.4 nM (IC50) PKCβI 8.1 nM (IC50) PKCβII 6.9 nM (IC50) PKCγ 45.9 nM (IC50) PKCδ 586 nM (IC50) PKCθ 29.5 nM (IC50) PKCε 522 nM (IC50)
PF-04577806 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 the development of diabetic microvascular abnormalities, making PKC a therapeutic target for diabetic complications. |
|---|---|
| ln Vitro |
In diabetic rat retinal lysates, PF-04577806 (0.001-10 μM; 10 min) inhibits PKC activity with an IC50 of 0.18 μM[1]. PF-04577806 (0.12–10 μM; pretreated for 60 min) inhibits the phosphorylation of SHP2 in HEK293 cells in response to phorbol myristate acetate stimulation, while PF-04577806 (0.001–10 μM; pretreated for 1 h) has an IC50 of 5.8 nM. With an IC50 of 0.12 μM, PF-04577806 (0.001-10 μM; pretreated for 1 h) concentration-dependently inhibits interleukin 8 release from phorbol myristate acetate-stimulated HEK293 cells[1]. When applied to human umbilical vein endothelial cells, PF-04577806 (1 μM; 48 h) exhibits minimal cytotoxicity, leaving 100.5% of the cells viable[1].
In vitro, PF-04577806 inhibits PKCα with an IC50 of 2.4 nM, PKCβI with 8.1 nM, PKCβII with 6.9 nM, PKCγ with 45.9 nM, and PKCθ with 29.5 nM. In diabetic rat retinal lysates stimulated with phorbol myristate acetate, PF-04577806 inhibits PKC activity with an IC50 of 0.18 µM within 10 minutes. The compound shows high specificity for PKC over other protein kinases. |
| ln Vivo |
In vivo, PF-04577806 demonstrates good oral exposure in the retina of diabetic Long-Evans rats. When orally administered, it ameliorates retinal vascular leakage in a streptozotocin-induced diabetic rat model. This efficacy is attributed to its ability to block PKC signaling in retinal tissue. The compound represents a promising new class of targeted kinase inhibitors with potential as therapeutic agents for patients with diabetic microvascular complications.
|
| Enzyme Assay |
For non-cell enzyme assays, recombinant PKC isoforms are incubated with PF-04577806 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.
|
| Cell Assay |
Western Blot Analysis[1]
Cell Types: Jurkat T cells Tested Concentrations: 0, 0.12, 0.37, 1.11, 3.33, 10.0 μM Incubation Duration: Pretreated for 1 hrs (hours) Experimental Results: demonstrated a dose-dependent decrease in phospho-ERK1/2 but not total ERK1/2. In cell-based functional assays, PF-04577806 is tested in multiple cellular systems. It blocks PKC signaling activity in a concentration-dependent manner. Specifically, it inhibits phospho-ERK1/2 formation, interleukin-8 release, and phospho-SHP2 levels. These cellular readouts confirm target engagement and functional inhibition of PKC-mediated signaling pathways. Low cytotoxicity is observed at 1 µM against human umbilical vein endothelial cells, with cell viability remaining at 83.3%. |
| Animal Protocol |
For in vivo animal studies, streptozotocin-induced diabetic Long-Evans rats are used. PF-04577806 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 exposure in the retina and ameliorates vascular leakage, demonstrating target engagement and efficacy in the diabetic retina.
|
| References |
| Molecular Formula |
C26H37N7O3
|
|---|---|
| Molecular Weight |
495.62
|
| Exact Mass |
495.295
|
| CAS # |
1072100-81-2
|
| PubChem CID |
25014995
|
| Appearance |
Typically exists as solid at room temperature
|
| Density |
1.2±0.1 g/cm3
|
| Boiling Point |
613.4±55.0 °C at 760 mmHg
|
| Flash Point |
324.8±31.5 °C
|
| Vapour Pressure |
0.0±1.8 mmHg at 25°C
|
| Index of Refraction |
1.602
|
| LogP |
2.02
|
| tPSA |
107
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
4
|
| Heavy Atom Count |
36
|
| Complexity |
804
|
| Defined Atom Stereocenter Count |
2
|
| SMILES |
O1CCC(CC1)CN1C[C@H](C)N(C(N2CC3C(NC(C4C=CC=CN=4)=O)=NNC=3C2(C)C)=O)C[C@H]1C
|
| InChi Key |
WIUOWTNXQICTJG-MSOLQXFVSA-N
|
| InChi Code |
InChI=1S/C26H37N7O3/c1-17-14-32(18(2)13-31(17)15-19-8-11-36-12-9-19)25(35)33-16-20-22(26(33,3)4)29-30-23(20)28-24(34)21-7-5-6-10-27-21/h5-7,10,17-19H,8-9,11-16H2,1-4H3,(H2,28,29,30,34)/t17-,18+/m1/s1
|
| Chemical Name |
N-[5-[(2S,5R)-2,5-dimethyl-4-(oxan-4-ylmethyl)piperazine-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 | 2.0177 mL | 10.0884 mL | 20.1767 mL | |
| 5 mM | 0.4035 mL | 2.0177 mL | 4.0353 mL | |
| 10 mM | 0.2018 mL | 1.0088 mL | 2.0177 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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