Pamapimod

Catalog No.S8125 Batch:S812501

Technical Data

Formula

C₁₉H₂₀F₂N₄O₄

Molecular Weight

406.38

CAS No.

449811-01-2

Solubility (25°C)*

In vitro

DMSO

81 mg/mL (199.32 mM)

Ethanol

28 mg/mL (68.9 mM)

Water

Insoluble

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

Clear solution	5% DMSO 1 95% Corn oil	0.405mg/ml	Taking the 1 mL working solution as an example, add 50 μL of 8.1 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results.
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O	4.05mg/ml	Taking the 1 mL working solution as an example, add 50 μL of 81 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 clarify; 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

Pamapimod (R-1503, Ro4402257) is a novel, selective inhibitor of p38 mitogen-activated protein kinase. It inhibits p38α and p38β enzymatic activity with IC50 values of 0.014±0.002 and 0.48± 0.04 microM, respectively with no activity against p38delta or p38gamma isoforms.

Targets

p38α ^[1] (Cell-free assay)	p38β ^[1] (Cell-free assay)
0.014 μM	0.48 μM

In vitro

Pamapimod inhibited p38, but inhibition of JNK was not detected. Pamapimod also inhibited lipopolysaccharide (LPS)-stimulated tumor necrosis factor (TNF) α production by monocytes, interleukin (IL)-1β production in human whole blood, and spontaneous TNFα production by synovial explants from RA patients^[1].

In vivo

In murine collagen-induced arthritis, pamapimod reduced clinical signs of inflammation and bone loss at 50 mg/kg or greater. In a rat model of hyperalgesia, pamapimod increased tolerance to pressure in a dose-dependent manner, suggesting an important role of p38 in pain associated with inflammation. Pamapimod suppresses spontaneous production of TNFα by synovial explants from RA patients. LPS- and TNFα-stimulated production of TNFα and IL-6 in rodents also was inhibited by pamapimod^[1].

Protocol (from reference)

Cell Assay:^{^[1]}	Cell lines Human Monocytic Cell Line, THP-1 Concentrations 15 μM Incubation Time 150 min Method THP-1 cells, growing in log phase, were collected by centrifugation and resuspended in RPMI 1640 containing 5.5×10^-5 M 2-mercaptoethanol and 10% fetal bovine serum to a final cell concentration of 2.5×10⁶ cells/ml. Dilutions of pamapimod were predispensed in 25 μl aliquots (before addition of cells) into round-bottom 96-well plates. The starting concentration was 100 μM in 5% dimethyl sulfoxide, and six half-log serial dilutions were made. After the addition of 200 μl of cell suspension and 25 μl of 5 μg/ml LPS in medium, the final dimethyl sulfoxide concentration was 0.5%. Compounds were diluted an additional 10-fold, and the final LPS concentration was 500 ng/ml. The cell suspensions and compound dilutions were combined and incubated for 30 min at 37°C in a 5% CO₂ humidified atmosphere, before the addition of LPS (or medium for non-LPS control samples). After the addition of LPS, plates were incubated for 2 h, followed by centrifugation to pellet cells. Cell supernatants were stored at 4°C until analysis for TNF-α content. TNF-α levels were determined by ELISA. Cytokine concentrations were determined.
Animal Study:^{^[1]}	Animal Models BALB/c mice Dosages 25, 50, 100, 150 mg/kg Administration oral administration

Cell Assay:^{^[1]}

Cell lines
Human Monocytic Cell Line, THP-1
Concentrations
15 μM
Incubation Time
150 min
Method
THP-1 cells, growing in log phase, were collected by centrifugation and resuspended in RPMI 1640 containing 5.5×10^-5 M 2-mercaptoethanol and 10% fetal bovine serum to a final cell concentration of 2.5×10⁶ cells/ml. Dilutions of pamapimod were predispensed in 25 μl aliquots (before addition of cells) into round-bottom 96-well plates. The starting concentration was 100 μM in 5% dimethyl sulfoxide, and six half-log serial dilutions were made. After the addition of 200 μl of cell suspension and 25 μl of 5 μg/ml LPS in medium, the final dimethyl sulfoxide concentration was 0.5%. Compounds were diluted an additional 10-fold, and the final LPS concentration was 500 ng/ml. The cell suspensions and compound dilutions were combined and incubated for 30 min at 37°C in a 5% CO₂ humidified atmosphere, before the addition of LPS (or medium for non-LPS control samples). After the addition of LPS, plates were incubated for 2 h, followed by centrifugation to pellet cells. Cell supernatants were stored at 4°C until analysis for TNF-α content. TNF-α levels were determined by ELISA. Cytokine concentrations were determined.

Animal Study:^{^[1]}

Animal Models
BALB/c mice
Dosages
25, 50, 100, 150 mg/kg
Administration
oral administration

References

[1] Hill RJ, et al. J Pharmacol Exp Ther. 2008, 327(3):610-9.

Selleck's Pamapimod has been cited by 6 publications

IL-33 promotes type 1 cytokine expression via p38 MAPK in human NK cells. [ J Leukoc Biol, 2020, 10.1002/JLB.3A0120-379RR]	PubMed: 32017227
p38α Regulates Expression of DUX4 in a Model of Facioscapulohumeral Muscular Dystrophy [ J Pharmacol Exp Ther, 2020, 374(3):489-498]	PubMed: 32576599
Corticosteroids inhibit Mycobacterium tuberculosis-induced necrotic host cell death by abrogating mitochondrial membrane permeability transition. [ Nat Commun, 2019, 10(1):688]	PubMed: 30737374
The Novel p38 Inhibitor, Pamapimod, Inhibits Osteoclastogenesis and Counteracts Estrogen-Dependent Bone Loss in Mice [ J Bone Miner Res, 2019, 34(5):911-922]	PubMed: 30615802
Clinically Advanced p38 Inhibitors Suppress DUX4 Expression in Cellular and Animal Models of Facioscapulohumeral Muscular Dystrophy. [ J Pharmacol Exp Ther, 2019, 370(2):219-230]	PubMed: 31189728
EGFR signaling suppresses type 1 cytokine-induced T-cell attracting chemokine secretion in head and neck cancer [ PLoS One, 2018, 13(9):e0203402]	PubMed: 30192802

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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.

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