Home MAPK p38 MAPK inhibitor Neflamapimod (VX-745)

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Neflamapimod (VX-745) p38 MAPK inhibitor

Neflamapimod (VX-745) is a potent and selective inhibitor of p38α with IC50 of 10 nM, and it shows 22-fold greater selectivity versus p38β and no inhibition to p38γ.

Neflamapimod (VX-745) p38 MAPK inhibitor Chemical Structure

Chemical Structure

Molecular Weight: 436.26

Purity & Quality Control

Related Products

Signaling Pathway

p38 MAPK Signaling Pathways

p38 MAPK Signaling Pathway

Mechanism of Action

Features A potent and selective inhibitor to p38α and p38β MAPK.
Targets
p38α [1] p38β [1]
10 nM 220 nM

In vitro
In vitro Neflamapimod (VX-745) selectively inhibits p38α and p38β MAPK with IC50 of 10 nM and 220 nM, respectively, but not p38γ MAPK and a large panel of other kinases, with IC50 larger than 20 µM. In a human peripheral blood mononuclear cell (PBMC) assay, it provides IC50 of 56 and 52 nM for IL-1β and TNFα, respectively. This compound blocks IL-6 and IL-8 production induced by IL-1 and TNFα, and COX-2 synthesis mediated by LPS and IL-1β. [1-3] It (60 nM-20 µM) inhibits IL-6 and VEGF secretion in bone marrow stromal cells (BMSCs), without affecting their viability. VX-745 also inhibits TNF-α-induced IL-6 secretion in BMSCs. It inhibits both multiple myeloma (MM) cell proliferation and IL-6 secretion in BMSCs triggered by adherence of MM cells to BMSCs, suggesting that it can inhibit paracrine multiple myeloma (MM) cell growth in the BM milieu and overcome cell adhesion-related drug resistance. [4]
Kinase Assay Spectrophotometric coupled-enzyme assay
Neflamapimod (VX-745) was evaluated for its inhibitory activity against p38α and p38β homologs using a spectrophotometric coupled-enzyme assay. A fixed concentration of enzyme (15 nM of p38α or p38β) was incubated with it in DMSO for 10 min. at 30 °C in 0.1 M HEPES buffer, pH 7.5, containing 10% glycerol, 10 mM MgCl2, 2.5 mM phosphoenolpyruvate, 200 µM NADH, 150 µg/mL pyruvate kinase, 50 µg/mL lactate dehydrogenase, and 200 µM EGF receptor peptide (KRELVEPLTPSGEAPNQALLR). The reaction was initiated with 100 µM and 70 µM ATP for p38α and p38β assays, respectively. The decrease of absorbance at 340 nm was monitored to follow the rate of the reaction. IC50 was evaluated from the rate data as a function of the inhibitor concentration.
Cell Research Cell lines Human bone marrow stromal cells (BMSCs) and multiple myeloma (MM) cells
Concentrations 60 nM - 20 μM, dissolved in DMSO.
Incubation Time 48 hours
Method Neflamapimod (VX-745) is used to incubate BMSCs (5 × 104 cells/well) or MM cells (3 × 104 cells/well) in 96-well culture plates for 48 hours at 37 °C, either in its presence or absence. DNA synthesis is measured by [3H]-thymidine ([3H]TdR) uptake. Cells are pulsed with [3H]TdR (0.5 μCi/well [.0185 MBq]) during the last 8 hours of 48-hour cultures. Growth inhibition of both MM cells and BMSCs by this compound is also assessed by measuring 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) dye absorbance.

In Vivo
In vivo Neflamapimod (VX-745) is effective against adjuvant-induced arthritis (AA) in the rat with ED50 of 5 mg/kg. Histological scores for this compound in AA rats are 93% inhibition of bone resorption and 56% inhibition of inflammation. In the classical cartilage-induced arthritis model, it exhibits a dose-responsive decrease in severity score. [1-3] In a type II collagen-induced arthritis (CIA) mice model, VX-745 (2.5, 5, and 10 mg/kg) has 27%, 31%, and 44% improvement in the inflammatory scores, respectively, when compared to vehicle-treated mice. In addition, histological scores show a 32-39% protection of bone and cartilage erosion by it. [5]
Animal Research Animal Models Type II collagen-induced arthritis (CIA) mice model (DBA/1J)
Dosages 2.5, 5, and 10 mg/kg
Administration Oral gavage twice daily
NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT02423200 Completed
Alzheimer''s Disease
EIP Pharma Inc
 April 2015 Phase 2

References
  • #
  • https://pubmed.ncbi.nlm.nih.gov/11892915/
  • #
  • https://pubmed.ncbi.nlm.nih.gov/12393542/
  • http://pubs.acs.org/doi/abs/10.1021/ml2001455

Chemical Information

Molecular Weight 436.26 Formula

C19H9Cl2F2N3OS
CAS No. 209410-46-8 SDF Download SDF
Synonyms N/A
Smiles C1=CC(=C(C(=C1)Cl)C2=C3C=CC(=NN3C=NC2=O)SC4=C(C=C(C=C4)F)F)Cl

Storage and Stability

Storage (From the date of receipt)

In vitro
Batch:

DMSO : 15 mg/mL ( (34.38 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Water : Insoluble

Ethanol : Insoluble


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In vivo
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Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

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