R59949

Catalog No.E0082 Batch:E008201

Technical Data

Formula

C₂₈H₂₅F₂N₃OS

Molecular Weight

489.58

CAS No.

120166-69-0

Solubility (25°C)*

In vitro

DMSO

98 mg/mL (200.17 mM)

Ethanol

2 mg/mL (4.08 mM)

Water

Insoluble

In vivo (Add solvents to the product individually and in order)

Clear solution

5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O

5.0mg/ml

Taking the 1 mL working solution as an example, add 50 μL of 100 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to make it clear; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.

* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description

R59949, a diacylglycerol kinase (DGK) inhibitor with an IC50 of 300 nM, inhibits inducible nitric oxide production through decreasing transplasmalemmal L-arginine uptake in vascular smooth muscle cells. R59949 strongly inhibits type I DGK a and γ, and moderately attenuates type II DGK θ and κ.

Targets

DKG ^[1]

0.3 μM

In vitro

R59949 is a pan diacylglycerol kinase (DGK) inhibitor, which inhibits inducible NO production through decreasing transplasmalemmal L-arginine uptake in rat aortic smooth muscle cells (RASMCs).^[1]

In vivo

R59949 upregulates the expression of PHD-2, also downregulates HIF-1α and vascular endothelial growth factor (VEGF) in the retina of oxygen-induced retinopathy (OIR) mice, suggesting a potential possibility that R59949 suppresses retinal neovascular pathophysiology via PHD2/HIF-1α/VEGF pathway.^[2]

Protocol (from reference)

Cell Assay:^{^[1]}	Cell lines rat aortic smooth muscle cells (RASMCs) Concentrations 10 µM Incubation Time 30 min Method RASMCs are prepared from adult male Wistar rats and cultured in DMEM containing 10% FBS under 5% CO2 at 37 ℃. Cultured cells showing a confluent condition in a 24-well plate are used for experiments. The cells are pretreated with R59949 at 10 μM or cycloheximide at 1 μM at 30 min before stimulation with IL-1β at 10 ng/ml or a vehicle for 24 h. Measurement of NOX is performed in the presence of nitrate reductase using a commercial kit. After incubation with nitrate reductase at room temperature for 2 h, aliquots of the culture medium are mixed with Griess reagent (1% sulphanilamide/0.1% N-(1-naphthyl) ethylenediamine dihydrochloride in 5% phosphoric acid). The mixture is then incubated at room temperature for 10 min, and its absorbance is measured at 540 nm using a photometer.
Animal Study:^{^[2]}	Animal Models C57BL/6J mice Dosages 10 μg/g/day Administration i.p.

Cell Assay:^{^[1]}

Cell lines
rat aortic smooth muscle cells (RASMCs)
Concentrations
10 µM
Incubation Time
30 min
Method
RASMCs are prepared from adult male Wistar rats and cultured in DMEM containing 10% FBS under 5% CO2 at 37 ℃. Cultured cells showing a confluent condition in a 24-well plate are used for experiments. The cells are pretreated with R59949 at 10 μM or cycloheximide at 1 μM at 30 min before stimulation with IL-1β at 10 ng/ml or a vehicle for 24 h. Measurement of NOX is performed in the presence of nitrate reductase using a commercial kit. After incubation with nitrate reductase at room temperature for 2 h, aliquots of the culture medium are mixed with Griess reagent (1% sulphanilamide/0.1% N-(1-naphthyl) ethylenediamine dihydrochloride in 5% phosphoric acid). The mixture is then incubated at room temperature for 10 min, and its absorbance is measured at 540 nm using a photometer.

Animal Study:^{^[2]}

Animal Models
C57BL/6J mice
Dosages
10 μg/g/day
Administration
i.p.

References

RETURN POLICY
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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

NOT FOR HUMAN, VETERINARY DIAGNOSTIC OR THERAPEUTIC USE.