1. Protein Tyrosine Kinase/RTK
  2. Src

WH-4-023 (Synonyms: Dual LCK/SRC inhibitor)

Cat. No.: HY-12299
Handling Instructions

WH-4-023 is a potent and selective dual Lck/Src inhibitor with IC50 of 2 nM/6 nM for Lck and Src kinase respectively; little inhibition on p38α and KDR.

For research use only. We do not sell to patients.
WH-4-023 Chemical Structure

WH-4-023 Chemical Structure

CAS No. : 837422-57-8

Size Price Stock Quantity
Free sample   Apply now  
10 mM * 1 mL in DMSO $100 In-stock
5 mg $80 In-stock
10 mg $120 In-stock
50 mg $400 In-stock
100 mg $650 In-stock
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500 mg   Get quote  

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Description

WH-4-023 is a potent and selective dual Lck/Src inhibitor with IC50 of 2 nM/6 nM for Lck and Src kinase respectively; little inhibition on p38α and KDR.

IC50 & Target

IC50: 2 nM (Lck), 6 nM (Src)[1]

In Vitro

WH-4-023 shows a similar potency increase on Lck as 2-substituted variants[1].

References
Preparing Stock Solutions
Concentration Volume (DMSO) Mass 1 mg 5 mg 10 mg
1 mM 1.7585 mL 8.7924 mL 17.5849 mL
5 mM 0.3517 mL 1.7585 mL 3.5170 mL
10 mM 0.1758 mL 0.8792 mL 1.7585 mL
Kinase Assay
[1]

The Lck HTRF kinase assay involves ATP-dependent phosphorylation of a biotinylated substrate peptide of gastrin in the presence or absence of inhibitor compound. The final concentration of gastrin is 1.2 μM. The final concentration of ATP is 0.5 μM (Km app =0.6±0.1 μM), and the final concentration of Lck (a GST-kinase domain fusion (AA 225−509)) is 250 pM. Buffer conditions are as follows: 50 mM HEPES pH=7.5, 50 mM NaCl, 20 mM MgCl2, 5 mM MnCl2, 2 mM DTT, 0.05% BSA. The assay is quenched and stopped with 160 μL of detection reagent. Detection reagents are as follows:  Buffer made of 50 mM Tris, pH=7.5, 100 mM NaCl, 3 mM EDTA, 0.05% BSA, 0.1% Tween20. Prior to reading, Streptavidin allophycocyanin (SA-APC) is added at a final concentration in the assay of 0.0004 mg/mL, along with europilated anti-phosphotyrosine Ab (Eu-anti-PY) at a final conc of 0.025 nM. The assay plate is read in a Discovery fluorescence plate reader with excitation at 320 nm and emission at 615 and 655 nm[1].

Cell Assay
[1]

WH-4-023 is dissolved in DMSO and stored, and then diluted with appropriate medium before use[1].

The purpose of this assay is to test the potency of T cell receptor (TCR; CD3) and CD28 signaling pathway inhibitors in human T cells. T cells are purified from human peripheral blood lymphocytes (hPBL) and preincubated with or without compound prior to stimulation with a combination of an anti-CD3 and an anti-CD28 antibody in 96-well tissue culture plates (1×105 T cells/well). Cells are cultured for ∼20 h at 37°C in 5% CO2 and then secreted IL-2 in the supernatants is quantified by cytokine ELISA. The cells remaining in the wells are then pulsed with 3H-thymidine overnight to assess the T cell proliferative response. Cells are harvested onto glass fiber filters and 3H-thymidine incorporation into DNA is analyzed by liquid scintillation counter. For comparison purposes, phorbol myristic acid (PMA) and calcium ionophore are used in combination to induce IL-2 secretion from purified T cells. Potential inhibitor compounds are tested for inhibition of this response as described above for anti-CD3 and -CD28 antibodies. Human whole-blood anti-CD3/CD28-induced IL-2 secretion assays are run in a similar fashion as described above using whole blood from normal volunteers diluted 50% in tissue culture medium prior to stimulation[1].

References
M.Wt

568.67

Formula

C₃₂H₃₆N₆O₄

CAS No.

837422-57-8

Storage
Powder -20°C 3 years
  4°C 2 years
In solvent -80°C 6 months
  -20°C 1 month
Shipping

Room temperature in continental US; may vary elsewhere

Solvent & Solubility

DMSO: ≥ 46.8 mg/mL

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WH-4-023
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HY-12299
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